To investigate the current distribution and predict the future suitable habitats of Haemaphysalis longicornis in Nanjing City, so as to provide insights into control and early warning of ticks and management of tick-borne diseases in Nanjing City. The electronic map of Nanjing City was obtained from the National Platform for Common GeoSpatial Information Services. The distribution of H. longicornis and the longitude and latitude of distribution points from 2022 to 2024 were obtained from centers for disease control and prevention across each district in Nanjing City. Climatic and environmental variable data in Nanjing City were captured from the Worldclim database. Initially, 19 bioclimatic variables in this database were selected, including annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the warmest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter. The elevation and normalized difference vegetation index were obtained from Data Sharing Platform of the Center for Resources and Environmental Sciences, Chinese Academy of Sciences. Then, the distribution points of H. longicornis, elevation, vegetation index and 19 bioclimatic variables were loaded into the software MaxEnt 3.4.4 to evaluate and screen out the variables with a contribution rate of 1% and higher. ArcGIS 10.8.1 software was used to extract the elevation, vegetation index and 19 bioclimatic variables of the distribution points of H. longicornis for a correlation analysis. If the absolute value of the correlation coefficient was 0.8 and higher, the variable with the higher contribution was retained. The 2050 dataset of the BCCCSM2-MR atmospheric circulation model in the coupled model intercomparison project phase 6 (CMIP6) were obtained from the Worldclim database as climate data for 2050. Screened H. longicornis species data and environmental and climate data were loaded into the maximum entropy (MaxEnt) model with the software MaxEnt 3.4.4 for training and validation, and then, all data generated from the model were imported into the software ArcGIS 10.8.1 to generate raster data and yield the map pertaining to the distribution of H. longicornis risk in Nanjing City. The accuracy of the model was evaluated with a receiver operating characteristic (ROC) curve, and the predictive effect of the model was assessed with area under the ROC curve (AUC). The suitable habitats of H. longicornis were classified in Nanjing City with the software ArcGIS 10.8.1, and the areas of distribution of suitable habitats in various categories were recorded to create the map of current H. longicornis suitable habitats classification in Nanjing City. The climatic and geographic information data in 2050 were employed as future environmental and climatic factors, and current environmental and climatic factors and current H. longicornis distribution data were additionally used to predict the future suitable habitats of H. longicornis in Nanjing City. In addition, the contributions of environmental and climatic factors to distribution of suitable habitats of H. longicornis was evaluated with the Jackknife method in Nanjing City. A total of 10 environmental and climatic variables were screened for analysis of the suitability of H. longicornis in Nanjing City based on correlation analyses and contributions of the MaxEnt model, including annual mean temperature, precipitation of the warmest quarter, vegetation index, precipitation of the wettest month, temperature annual range, annual precipitation, mean temperature of the warmest quarter, elevation, mean temperature of the wettest quarter, and maximum temperature of the warmest month, and annual mean temperature (34.8%), precipitation of the warmest quarter (17.3%), vegetation index (13.1%), and precipitation of the wettest month (10.8%) contributed relatively highly to the distribution of suitable habitats of H. longicornis in Nanjing City. The mean AUC of the ROC curve was 0.810 ± 0.055 for 10 repeated modeling results of the MaxEnt model, indicating high predictive performance of the model. The potential distribution areas of H. longicornis were predicted to be mainly located in Luhe District, Pukou District, Jiangning District, Lishui District, and Gaochun District in Nanjing City with the MaxEnt model. Under current climatic conditions, the area of potential suitable habitats of H. longicornis was 4 182.42 km2 in Nanjing City, including 1 252.94 km2 highly suitable habitats, which accounted for 19.00% of the total area of Nanjing City. Under the climate scenario in 2050, the area of potential suitable habitats of H. longicornis was projected to increase to 5 467.58 km2 in Nanjing City, accounting for 82.95% of the total area of the city, and these habitats were mainly concentrated in Luhe District, Pukou District, Jiangning District, Lishui District, and Gaochun District. The areas of suitable habitats of H. longicornis at various categories were predicted to vary greatly in 2050, and the area of highly suitable habitats of H. longicornis was projected to increase to 2 378.82 km2, accounting for 36.08% of the total area of Nanjing City. Based on jackknife tests and contributions of environmental and climatic variables, 6 dominant environmental and climatic factors were screened, including annual mean temperature (34.8% contribution), precipitation of the warmest quarter (17.3% contribution), vegetation index (13.1% contribution), precipitation of the wettest month (10.8% contribution), temperature annual range (5.4% contribution), and mean temperature of the warmest quarter (5.0% contribution), with cumulative contributions of 86.4%. The distribution of H. longicornis is strongly associated with vegetation, temperature and precipitation in Nanjing City. Future climate change may lead to an expansion of the distribution area of H. longicornis in Nanjing City. ［摘要］ 目的 分析南京市长角血蜱 (Haemaphysalis longicornis) 分布现状并预测其未来适生区范围, 为该市蜱媒控制预 警及蜱传疾病防治提供参考。方法 于国家地理信息公共服务平台获取南京市电子地图。于南京市各区疾病预防控制 中心获取2022—2024年各区长角血蜱分布数据及分布点经纬度信息。于世界气候数据库获取该期间南京市气候环境 变量数据, 初步选取该数据库中全部19个生物气候变量, 包括年平均气温、昼夜温差月均值、等温性、温度季节性变化、最暖月份最高气温、最暖月份最低气温、平均年温差、最湿季度平均温度、最干季度平均温度、最暖季度平均温度、最冷季 度平均温度、年降水量、最湿月份降水量、最干月份降水量、季节性降水量、最湿季度降水量、最干季度降水量、最暖季度 降水量和最冷季度降水量。于中国科学资源环境数据中心共享平台获取海拔、归一化植被指数。将长角血蜱分布点、海 拔、植被指数和19个生物气候变量数据导入MaxEnt 3.4.4软件, 评估并筛选出贡献率≥ 1%的变量。采用ArcGIS 10.8.1软 件提取长角血蜱分布点海拔、植被指数和上述19个生物气候变量数据并进行相关性分析, 当相关系数绝对值≥ 0.8时, 保 留两者中贡献率较高者。在世界气候数据库中, 选取第六次国际耦合模式比较计划中的BCCCSM2-MR大气环流模型 2050年数据集作为2050年气候数据。采用MaxEnt 3.4.4软件, 将筛选所得长角血蜱物种数据与环境气候数据导入最大 熵 (maximum entropy, MaxEnt) 模型进行训练和验证, 将模型输出结果导入ArcGIS 10.8.1软件形成栅格数据, 得到南京市 长角血蜱风险分布图。采用受试者工作特征 (receiver operating characteristic, ROC) 曲线验证模型准确性, 并以ROC曲线 下面积 (area under curve, AUC) 评估预测效果。采用ArcGIS 10.8.1软件对南京市长角血蜱适生区进行等级划分, 统计不 同等级适生区分布面积, 绘制当前南京市长角血蜱适生区等级图。以2050年气候与地理信息数据作为未来环境气候因 子, 结合当前环境气候因子及长角血蜱分布数据预测未来南京市长角血蜱适生区。采用刀切法评估各环境气候因子对 南京市长角血蜱适生区分布的影响。结果 结合相关性分析与MaxEnt模型贡献率, 筛选出年平均气温、最暖季度降水 量、植被指数、最湿月份降水量、平均年温差、年降水量、最暖季度平均温度、海拔、最湿季度平均温度和最暖月份最高气 温10个环境气候变量用于南京市长角血蜱适生性分析, 其中年平均气温、最暖季度降水量、植被指数和最湿月份降水量 贡献率较高, 分别为34.8%、17.3%、13.1%和10.8%。10次重复建立的MaxEnt模型ROC曲线AUC均值为0.810 ± 0.055, 模型预测性能较好。MaxEnt模型预测结果显示, 南京市长角血蜱潜在分布区主要位于六合区、浦口区、江宁区、溧水区 和高淳区。当前气候条件下, 南京市长角血蜱潜在适生区面积为4 182.42 km2; 其中高适生区面积为1 252.94 km2, 占南 京市总面积的19.00%。在未来2050年气候情景下, 南京市长角血蜱潜在适生区面积将增至5 467.58 km2, 占全市总面积 的82.95%; 适生区主要集中于六合区、浦口区、江宁区、溧水区与高淳区; 不同等级适生区面积较当前变化较大, 其中高 适生区面积增至2 378.82 km2, 占南京市总面积的36.08%。综合刀切法检验和环境气候变量贡献率, 筛选出年平均气温 (贡献率34.8%)、最暖季度降水量 (贡献率17.3%)、植被指数 (贡献率13.1%)、最湿月份降水量 (贡献率10.8%)、平均年温 差 (贡献率5.4%)、最暖季度平均温度 (贡献率5.0%) 6个主导环境气候因子, 累计贡献率为86.4%。结论 南京市长角血 蜱分布与植被、气温和降水量密切相关。未来气候变化可能导致南京市长角血蜱分布区域扩大。.
To investigate the effectiveness of the integrated schistosomiasis control programme in Sichuan Province during the stage moving from transmission interruption to elimination (2015-2023), so as to provide insights into formulation of the schistosomiasis control measures during the post-elimination stage. Schistosomiasis control data were retrospectively collected from departments of health, agriculture and rural affairs, forestry and grassland, water resources, and natural resources in Sichuan Province from 2015 to 2023, and a database was created to document examinations and treatments of human and livestock schistosomiasis, and snail survey and control, conversion of paddy fields to dry fields, ditch hardening, rivers and lakes management and building of forests for snail control and schistosomiasis prevention. The completion of schistosomiasis control measures was investigated, and the effectiveness was evaluated. A total of 20 545 155 person-times received human schistosomiasis examinations in Sichuan Province during the period from 2015 to 2023, and 232 157 person-times were seropositive, with a reduction in the seroprevalence from 2.10% (44 299/2 107 003) in 2015 to 1.12% (9 361/837 896) in 2023 (χ2 = 7.68, P < 0.001). The seroprevalence of human schistosomiasis appeared a tendency towards a decline in Sichuan Province over years from 2015 to 2023 (b = -8.375, t = -10.052, P < 0.001); however, no egg positive individuals were identified during the period from 2018 to 2023, with the prevalence of human Schistosoma japonicum infections maintained at 0. Expanded chemotherapy was administered to 2 754 515 person-times, and medical assistance of advanced schistosomiasis was given to 6 436 persontimes, with the treatment coverage increasing from 46.80% (827/1 767) in 2015 to 64.87% (868/1 338) in 2023. Parasitological tests for livestock schistosomiasis were performed in 35 113 herd-times, and expanded chemotherapy was administered to 513 043 herd-times, while the number of fenced livestock decreased from 121 631 in 2015 to 103 489 in 2023, with a reduction of 14.92%. Snail survey covered 433 621.80 hm2 in Sichuan Province from 2015 to 2023, with 204 602.81 hm2 treated by chemical control and 4 637.74 hm2 by environmental modifications. The area of snail habitats decreased from the peak of 5 029.80 hm2 in 2016 to 3 709.72 hm2 in 2023, and the actual area of snail habitats decreased from the peak of 8 585.48 hm2 in 2016 to 473.09 hm2 in 2023. The mean density of living snails remained low across the study period except in 2017 (0.62 snails/0.1 m2). Schistosomiasis control efforts by departments of agriculture and rural affairs in Sichuan Province included conversion of paddy fields to dry fields covering 153 346.93 hm2, hardening of 6 110.31 km ditches, building of 70 356 biogas digesters, replacement of cattle with 227 161 sets of machines, and captive breeding of 21 161 070 livestock from 2015 to 2023, and the control efforts by departments of water resources included rivers and lakes management measuring 5 676.92 km and renovation of 2 331 irrigation areas, while the control efforts by departments of forestry and grassland included building of forests for snail control and schistosomiasis prevention covering 23 913.33 hm2, renovation of snail control forests covering 8 720 hm2 and newly building of shelterbelts covering 764 686.67 hm2. All 63 endemic counties (cities and districts) had achieved the criterion for schistosomiasis elimination criteria in Sichuan Province by the end of 2023. Following the integrated control efforts from 2015 to 2023, remarkable achievements have been obtained in the schistosomiasis control programme in Sichuan Province, with all endemic counties successfully attaining the schistosomiasis elimination target at the county level. ［摘要］ 目的 分析 2015—2023 年四川省血吸虫病从传播阻断到消除阶段的综合防治效果, 为制定消除后血吸虫病防 治措施提供科学依据。方法 收集 2015—2023 年四川省卫生健康、农业农村、林草、水利、自然资源等部门血吸虫病防 治相关资料, 统计人群、家畜查治病和流行区查灭螺, 以及水田改旱、沟渠硬化、河流湖泊治理、抑螺防病林建设等数据, 分析各项血吸虫病防治措施完成情况, 并评价防治效果。结果 2015—2023 年四川省累计开展人群查病 20 545 155 人· 次, 血检阳性 232 157 人·次, 血检阳性率从2015年的2.10% (44 299/2 107 003) 下降至 2023 年的 1.12% (9 361/837 896), 差异有统计学意义 (χ2 = 7.68, P < 0.001); 各年血检阳性率呈逐年下降趋势 (b = −8.375, t = −10.052, P < 0.001)。2018—2023 年均未发现病原学阳性病人, 人群血吸虫感染率持续为 0。9 年来共开展人群扩大化疗 2 754 515 人·次, 救治晚 期血吸虫病病人 6 436 人·次, 救治人数占比从 2015 年的 46.80% (827/1 767) 上升至 2023 年的 64.87% (868/1 338); 共开 展家畜病原学检查35 113头·次, 完成扩大化治疗513 043头·次, 家畜存栏量从2015年的121 631头降至 2023 年的 103 489 头, 下降了 14.92%。2015—2023 年四川省累计查螺 433 621.80 hm2, 药物灭螺 204 602.81 hm2, 环境改造灭螺 4 637.74 hm2, 查出有螺面积从 2016 年的 5 029.80 hm2 下降至 2023 年的 3 709.72 hm2, 实有钉螺面积从2019年的8 585.48 hm2 下降 至 2023 年的 473.09 hm2; 2017 年活螺平均密度为0.62只/0.1 m2, 其余年份均在较低水平波动。2015—2023 年四川省农 业部门累计完成水田改旱153 346.93 hm2, 沟渠硬化 6 110.31 km, 沼气池建设70 356 座, 以机代牛 227 161台 (套), 家畜 圈养21 161 070头 (只); 水利部门累计开展河流湖泊治理 5 676.92 km, 灌区改造 2 331处; 林草部门累计建设抑螺防病林 23 913.33 hm2, 抑螺林改造 8 720 hm2, 新建防护林 764 686.67 hm2。截至 2023 年底, 全省 63 个流行县 (市、区) 全部达到血 吸虫病消除标准。结论 2015—2023 年四川省实施各项血吸虫病综合防治措施后, 全省血吸虫病防治效果显著, 以县 为单位实现了消除血吸虫病目标。.
To investigate the antibacterial activity of the antifungal peptide Mt6-21DLeu derived from Musca domestica against Acinetobacter baumannii (AB) and unravel its underlying mechanisms, so as to provide insights into development of novel agents against AB. The minimum inhibitory concentrations (MICs) of Mt6-21DLeu, M. domestica-derived antifungal peptide-1 (MAF-1A), and polymyxin B were determined against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and AB using the broth microdilution assay, and the antibacterial activity of Mt6-21DLeu and polymyxin B was dynamically assessed against AB over 24 hours with time-kill curves. The inhibitory effects of Mt6-21DLeu and polymyxin B on biofilm formation in AB at concentrations of 1/4 × MIC, 1/2 × MIC and MIC, and the eradication effects of Mt6-21DLeu and polymyxin B on mature biofilms in AB at concentrations of MIC, 2 × MIC, and 4 × MIC were evaluated using crystal violet staining. Structural changes in the cell membrane of AB were observed 3 hours post-exposure to Mt6-21DLeu at concentrations of MIC and 2 × MIC using scanning electron microscopy, and alterations in the cell membrane permeability of AB were analyzed 3 hours post-treatment with Mt6-21DLeu at concentrations of MIC and 2 × MIC by means of fluorescence microscopy and propidium iodide (PI) staining. Intracellular reactive oxygen species (ROS) levels in AB were measured 3 hours post-treatment with Mt6-21DLeu at concentrations of MIC, 2 × MIC, and 4 × MIC using flow cytometry. The survival of Caenorhabditis elegans exposed to Mt6-21DLeu at concentrations of MIC, 2 × MIC, and 4 × MIC was monitored for 7 consecutive days, and survival curves were plotted to evaluate the in vivo toxicity of Mt6-21DLeu. In addition, C. elegans infected with AB and treated with Mt6-21DLeu at a concentration of 4 × MIC served as the treatment group, and uninfected C. elegans served as the control group, while infected but untreated C. elegans served as the infection group. The in vivo antibacterial efficacy of Mt6-21DLeu at a concentration of 4 × MIC was evaluated by comparing the survival curves and bacterial load among the three groups. The MICs of MAF-1A were all >128 μg/mL against S. aureus, B. subtilis, E. coli, K. pneumoniae, P. aeruginosa, and AB. In contrast, the MICs of Mt6-21DLeu were >128, 32, 8, 8, 16, and 4 μg/mL against these strains, respectively, and the MIC of Mt6-21DLeu against AB was close to that of polymyxin B (2 μg/mL). Time-kill curve analysis showed that both Mt6-21DLeu at concentrations of MIC and 2 × MIC and polymyxin B at a concentration of MIC inhibited AB growth over the 24-hour study period. The biofilm biomass in AB was (52.38 ± 6.92)%, (40.88 ± 9.17)% and (14.77 ± 6.00)% post-exposure with Mt6-21DLeu at concentrations of 1/4 × MIC, 1/2 × MIC and MIC, (61.58 ± 7.35)%, (47.42 ± 5.51)% and (20.85 ± 10.48)% post-treatment with polymyxin B at concentrations of 1/4 × MIC, 1/2 × MIC and MIC, and (100.00 ± 15.92)% in the control group (only bacterial suspension), respectively (F = 68.38, P < 0.001), and pairwise comparisons indicated that Mt6-21DLeu and polymyxin B at all concentrations significantly inhibited biofilm formation as compared to the control group (all P values < 0.001). The mature biofilm biomass in AB was (73.44 ± 11.41)%, (72.56 ± 13.08)% and (49.65 ± 9.23)% post-exposure to Mt6-21DLeu at concentrations of MIC, 2 × MIC, and 4 × MIC, (84.38 ± 8.60)%, (72.31 ± 9.63)% and (58.85 ± 4.96)% post-treatment with polymyxin B at concentrations of MIC, 2 × MIC, and 4 × MIC, and (100.00 ± 6.36)% in the control group (F = 35.63, P < 0.001), and pairwise comparisons revealed that Mt6-21DLeu at all concentrations significantly eradicated biofilm biomass (all P values < 0.05); however, polymyxin B showed no clear-cut eradication effect at a concentration of MIC (P > 0.05). Scanning electron microscopy revealed pore formation and content leakage in the cell membrane of AB 3 hours post-treatment with Mt6-21DLeu at concentrations of MIC and 2 × MIC. Fluorescence microscopy showed that the proportions of PI-stained AB were (24.79 ± 11.51)% and (68.44 ± 15.80)% post-treatment with Mt6-21DLeu at concentrations of MIC and 2 × MIC, and (0.96 ± 0.94)% in the phosphate-buffered saline (PBS) treatment group (F = 105.90, P < 0.001), with the highest proportion of PI-stained AB seen post-treatment with Mt6-21DLeu at a concentration of 2 × MIC (P < 0.05). Flow cytometry revealed that the relative intracellular ROS levels in AB were (652.00 ± 141.90), (694.33 ± 14.19), and (974.33 ± 160.02) 3 hours post-treatment with Mt6-21DLeu at concentrations of MIC, 2 × MIC and 4 × MIC, and (403.67 ± 86.56) in the PBS treatment group, respectively (F = 12.27, P < 0.05), with the highest intracellular ROS level measured following treatment with Mt6-21DLeu at a concentration of 4 × MIC (P < 0.05). Survival curve analysis revealed that Mt6-21DLeu posed no impact on C. elegans survival at concentrations of MIC (χ2 = 0.02, P > 0.05), 2 × MIC (χ2 = 0.06, P > 0.05) or 4 × MIC (χ2 = 0.16, P > 0.05), and there was a significant difference in the survival period of C. elegans among the control group, the infection group, and the treatment group (χ2 = 82.66, P < 0.05), with a significantly longer survival period in the treatment group than in the infection group (χ2 = 45.00, P < 0.05). In addition, the log-transformed bacterial colony counts in C. elegans were (0.00 ± 0.00), (5.46 ± 0.03), and (3.91 ± 0.47) CFU/mL in the control group, the infection group, and the treatment group, respectively (F = 324.80, P < 0.001), and the log-transformed bacterial colony counts in C. elegans were significantly lower in the treatment group than in the infection group (P < 0.05). Mt6-21DLeu exerts potent antibacterial effects through disrupting the cell membrane integrity of AB and promoting intracellular ROS accumulation in AB, and exhibits promising potential for treatment of AB infections both in vivo and in vitro, which may serve as a candidate drug molecule against multidrug-resistant AB infections. ［摘要］ 目的 探究家蝇抗真菌衍生肽Mt6-21DLeu对鲍曼不动杆菌的抗菌活性及其潜在机制, 为开发新型抗鲍曼不动 杆菌药物提供实验依据。方法 采用微量液体稀释法检测Mt6-21DLeu、家蝇抗真菌肽-1A (Musca domestica antifun-gal peptide-1A, MAF-1A) 、多黏菌素B对金黄色葡萄球菌、枯草芽孢杆菌、大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌、 鲍曼不动杆菌的最小抑菌浓度 (minimum inhibitory concentration, MIC), 通过时间-杀菌曲线动态评估Mt6-21DLeu和多黏 菌素B在24 h内对鲍曼不动杆菌的抑菌效果, 采用结晶紫染色法分别检测浓度为1/4 × MIC、1/2 × MIC、MIC的Mt6-21DLeu和多黏菌素B对鲍曼不动杆菌生物被膜的抑制作用以及浓度为MIC、2 × MIC、4 × MIC的Mt6-21DLeu和多黏菌素 B对鲍曼不动杆菌成熟生物被膜的清除效果。采用扫描电镜观察经浓度为MIC、2 × MIC的Mt6-21DLeu作用3 h时鲍曼 不动杆菌细胞膜结构变化, 采用荧光显微镜结合碘化丙啶 (propidium iodide, PI) 染色分析浓度为MIC、2 × MIC的Mt6-21DLeu作用3 h后鲍曼不动杆菌细胞膜通透性的变化, 采用流式细胞术检测浓度为MIC、2 × MIC、4 × MIC的Mt6-21DLeu 作用于鲍曼不动杆菌菌体3 h时其细胞内活性氧 (reactive oxygen species, ROS) 水平。此外, 连续7 d监测经浓度为MIC、 2 × MIC、4 × MIC的Mt6-21DLeu作用后秀丽隐杆线虫的生存状态, 绘制生存曲线评估Mt6-21DLeu的体内毒性。以感染 鲍曼不动杆菌且以浓度为4 × MIC的Mt6-21DLeu治疗的秀丽隐杆线虫为治疗组, 未感染鲍曼不动杆菌的秀丽隐杆线虫 为对照组, 感染鲍曼不动杆菌但未予治疗的秀丽隐杆线虫为感染组, 通过比较各组秀丽隐杆线虫生存曲线及体内带菌量 评估浓度为4 × MIC的Mt6-21DLeu的体内抗菌效果。结果 MAF-1A对金黄色葡萄球菌、枯草芽孢杆菌、大肠埃希菌、肺 炎克雷伯菌、铜绿假单胞菌、鲍曼不动杆菌的MIC均> 128 μg/mL, Mt6-21DLeu对上述各菌的MIC依次为> 128、32、8、8、 16、4 μg /mL, 其对鲍曼不动杆菌的MIC接近多黏菌素B (2 μg/mL) 。时间-杀菌曲线分析显示, 浓度为MIC、2 × MIC的Mt6-21DLeu和浓度为MIC的多黏菌素B均可在24 h内抑制鲍曼不动杆菌生长。经浓度为1/4 × MIC、1/2 × MIC、MIC的Mt6-21DLeu和多黏菌素B处理后, 鲍曼不动杆菌生物被膜含量依次为 (52.38 ± 6.92) %、 (40.88 ± 9.17) %、 (14.77 ± 6.00) %, (61.58 ± 7.35) %、 (47.42 ± 5.51) %和 (20.85 ± 10.48) %; 仅含鲍曼不动杆菌悬液的生长对照组生物被膜含量为 (100.00 ± 15.92) %; 各组差异有统计学意义 (F = 68.38, P < 0.001); 两两比较结果显示, 不同浓度的Mt6-21DLeu和多黏菌素B均可 抑制鲍曼不动杆菌生物被膜形成 (P 均< 0.001) 。此外, 浓度为MIC、2 × MIC、4 × MIC的Mt6-21DLeu和多黏菌素B处理 后, 鲍曼不动杆菌生物被膜含量依次为 (73.44 ± 11.41) %、 (72.56 ± 13.08) %、 (49.65 ± 9.23) %, (84.38 ± 8.60) %、 (72.31 ± 9.63) %、 (58.85 ± 4.96) %, 仅含鲍曼不动杆菌悬液的生长对照组生物被膜含量为 (100.00 ± 6.36) %, 差异有统计学意义 (F = 35.63, P < 0.001); 两两比较结果显示, 不同浓度的Mt6-21DLeu均可清除鲍曼不动杆菌生物被膜 (P 均< 0.05), 但浓 度为MIC的多黏菌素B对鲍曼不动杆菌生物被膜无显著清除作用 (P > 0.05) 。扫描电镜观察显示, 浓度为MIC及2 × MIC 的Mt6-21DLeu作用3 h后, 鲍曼不动杆菌细胞膜出现孔洞及内容物泄漏; 荧光显微镜观察显示, 经浓度为MIC及2 × MIC 的Mt6-21DLeu处理后PI染色的鲍曼不动杆菌比例分别为 (24.79 ± 11.51) %和 (68.44 ± 15.80) %, 而仅加入磷酸缓冲盐溶 液 (phosphate-buffered saline, PBS) 的鲍曼不动杆菌悬液中PI染色的鲍曼不动杆菌细胞比例为 (0.96 ± 0.94) %, 差异有统 计学意义 (F = 105.90, P < 0.001); 其中经浓度为2 × MIC的Mt6-21DLeu处理后PI染色的鲍曼不动杆菌细胞比例最高 (P < 0.05) 。流式细胞术检测结果显示, 浓度为MIC、2 × MIC、4 × MIC的Mt6-21DLeu作用于鲍曼不动杆菌菌体3 h时, 鲍 曼不动杆菌细胞内ROS相对丰度为 (652.00 ± 141.90) 、 (694.33 ± 14.19) 、 (974.33 ± 160.02), 而仅加入PBS的鲍曼不动杆 菌菌液中ROS相对丰度为 (403.67 ± 86.56), 差异有统计学意义 (F = 12.27, P < 0.05); 经浓度为4 × MIC的Mt6-21DLeu处 理后鲍曼不动杆菌细胞内ROS相对丰度最高 (P 均< 0.05) 。生存曲线分析结果显示, 浓度为MIC、2 × MIC、4 × MIC的 Mt6-21DLeu对秀丽隐杆线虫寿命无影响 (χ2 = 0.02、0.06、0.16, P > 0.05); 对照组、感染组、治疗组秀丽隐杆线虫生存时间差异有统计学意义 (χ2 = 82.66, P < 0.05), 其中治疗组较感染组生存时间延长 (χ2 = 45.00, P < 0.05) 。3组秀丽隐杆线虫 菌落浓度对数值分别为 (0.00 ± 0.00) 、 (5.46 ± 0.03) 、 (3.91 ± 0.47) CFU/mL, 差异有统计学意义 (F = 324.80, P < 0.001); 治 疗组线虫体内带菌量较感染组减少 (P < 0.05) 。结论 Mt6-21DLeu可通过破坏鲍曼不动杆菌细胞膜完整性、促进细胞内 ROS蓄积发挥高效抗菌作用, 且在体内外均表现出鲍曼不动杆菌感染治疗潜力, 可作为一种抗多重耐药鲍曼不动杆菌感 染的候选药物分子。.
To investigate the species of sandflies and the prevalence of Leishmania infections in sandflies from selected areas of northern and northwestern China, so as to provide insights into identification of leishmaniasis vectors and assessment of epidemiological trends of leishmaniasis in China. Sandfly samples were collected from Mentougou District of Beijing Municipality, Xiangning County in Linfen City of Shanxi Province, Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, and Payzawat County of Karamay District of Karamay City, Gaochang District of Turpan City in Xinjiang Uygur Autonomous Region from July 2023 to July 2024. Approximately 100 intact female sandfly samples were randomly selected from each site and the species of sandflies was identified according to morphological characteristics and molecular assays. Female sandflies originating from the same habitat were grouped into pools of 10 individuals. Leishmania infection was detected using polymerase chain reaction (PCR) assay targeting the internal transcribed spacer 1 (ITS-1) gene, and the prevalence of Leishmania infection was calculated in sandflies from different sampling sites using the minimum infection rate (MIR) method. In addition, positive amplicons were sequenced and subjected to phylogenetic analysis. A total of 6 155 sandflies were collected from different environments at sampling sites across the six aforementioned regions from July 2023 to July 2024. Phlebotomus chinensis (96.00%) was the dominant sandfly species in Mentougou District, Beijing Municipality, with a small proportion of Ph. sergenti (4.00%), and only Ph. chinensis was found in Xiangning County, Linfen City, Shanxi Province. Ph. wui was the only sandfly species detected in Ejin Banner, Alxa League, Inner Mongolia Autonomous Region, and Payzawat County, Kashgar City, Xinjiang Uygur Autonomous Region, and Ph. caucasicus (97.70%) was the dominant sandfly species in Karamay District, Karamay City, Xinjiang Uygur Autonomous Region, with a small proportion of Ph. wui (2.30%), while Ph. alexandri was the only species in Gaochang District, Turpan City, Xinjiang Uygur Autonomous Region. A total of 40, 60, 34, 18, 18, and 22 pools of sandfly samples were tested from Mentougou District in Beijing Municipality, Xiangning County in Linfen City of Shanxi Province, Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, Payzawat County in Kashgar City, Karamay District in Karamay City, and Gaochang District in Turpan City of Xinjiang Uygur Autonomous Region, respectively. L. infantum was detected in Ph. chinensis samples from Mentougou District in Beijing Municipality, and Xiangning County of Linfen City in Shanxi Province, with MIR of 0.25% to 1.00%, and L. donovani was detected in Ph. wui from Ejin Banner in Alxa League of Inner Mongolia Autonomous Region, and Payzawat County in Kashgar City of Xinjiang Uygur Autonomous Region, with MIR of 0.56% to 0.88%; however, no Leishmania infection was detected in Ph. caucasicus from Karamay District in Karamay City or Ph. alexandri from Gaochang District in Turpan City of Xinjiang Uygur Autonomous Region. Phylogenetic analysis showed that the Leishmania ITS-1 gene sequences obtained from Mentougou District in Beijing Municipality and Xiangning County in Linfen City of Shanxi Province were clustered into the same clade with the reference sequences of L. infantum ITS-1 gene, while the Leishmania ITS-1 gene sequences obtained from Ejin Banner in Alxa League of Inner Mongolia Autonomous Region and Payzawat County in Kashgar City of Xinjiang Uygur Autonomous Region were clustered into the same clade with the reference sequences of L. donovani ITS-1 gene. There are variations in sandfly species in selected areas of northern and northwestern China, and variations in the species of Leishmania infecting sandflies. Improved surveillance of sandfly vectors and targeted control strategies with adaptations to geographical features and leishmaniasis vectors are recommended. ［摘要］ 目的 调查我国华北及西北部分地区的媒介白蛉种类并检测其利什曼原虫感染情况, 为我国利什曼病传播媒 介识别与流行趋势判断提供科学依据。方法 2023年7月—2024年7月在北京市门头沟区、山西省临汾市乡宁县、内蒙 古自治区阿拉善盟额济纳旗、新疆维吾尔自治区喀什市伽师县、克拉玛依市克拉玛依区和吐鲁番市高昌区采集白蛉样 本, 并分别随机挑选完整的单只雌性白蛉样本约100只, 通过形态学与分子生物学方法鉴定确定白蛉蛉种。在各采样点 采集的白蛉样本中, 选取来源于同一种生境的雌性白蛉, 每10只分为1组; 以利什曼原虫内转录间隔区1 (internal transcribed spacer 1, ITS-1) 为目的基因, 采用PCR法检测上述地区各组白蛉利什曼原虫感染情况。利用最小感染率 (minimum infection rate, MIR) 法计算不同采样点白蛉的利什曼原虫感染率。此外, 扩增的阳性片段经测序后进行系统发育分 析。结果 2023年7月—2024年7月, 共在上述6个地区采样点的不同环境采集白蛉样本6 155只。经鉴定, 北京市门头 沟区采集的100只白蛉样本主要为中华白蛉 (96.00%), 其间混杂少许鳞喙司蛉 (4.00%); 山西省临汾市乡宁县采集的100 只白蛉样本均为中华白蛉; 内蒙古自治区阿拉善盟额济纳旗、新疆维吾尔自治区喀什市伽师县采集的100只白蛉样本均 为吴氏白蛉; 新疆维吾尔自治区克拉玛依市克拉玛依区采集的100只白蛉样本主要为高加索白蛉 (97.70%), 其间混杂少 许吴氏白蛉 (2.30%); 新疆维吾尔自治区吐鲁番市高昌区采集的100只白蛉样本均为亚历山大白蛉。北京市门头沟区、山西省临汾市乡宁县、内蒙古自治区阿拉善盟额济纳旗、新疆维吾尔自治区喀什市伽师县、克拉玛依市克拉玛依区和吐 鲁番市高昌区分别检测40、60、34、18、18、22组白蛉样本, 其中北京市门头沟区、山西省临汾市乡宁县中华白蛉样本中检 出婴儿利什曼原虫, MIR在0.25%～1.00%; 内蒙古自治区阿拉善盟额济纳旗、新疆维吾尔自治区喀什市伽师县吴氏白蛉 样本中检出杜氏利什曼原虫, MIR在0.56%～0.88%; 新疆维吾尔自治区克拉玛依市克拉玛依区高加索白蛉、吐鲁番市高 昌区亚历山大白蛉均未检出利什曼原虫。系统进化分析显示, 北京市门头沟区、山西省临汾市乡宁县采样点所获序列与 婴儿利什曼原虫参考序列聚为一支, 内蒙古自治区阿拉善盟额济纳旗、新疆维吾尔自治区喀什市伽师县采样点所获序列 与杜氏利什曼原虫参考序列聚为一支。结论 我国华北及西北部分地区白蛉种类存在差异, 且感染利什曼原虫虫种不 同。建议加强上述地区媒介白蛉监测, 并结合当地地理环境和传播媒介制定针对性的防控策略。.
To analyze Chinese and English publications pertaining to Oncomelania hupensis control from 2005 to 2024, so as to decipher the research status and hotspots of O. hupensis control. Chinese and English publications pertaining to O. hupensis control from 2005 to 2024 were retrieved in the Web of Science Core Collection Database and China National Knowledge Infrastructure. The annual number of publications was analyzed from 2005 to 2024, and the author and institution cooperation networks were mapped using the software CiteSpace 6.3.1. Keywords were extracted from publications to map the co-occurrence, burst and timeline of keywords to identify the research hotspots of O. hupensis control. A total of 158 English publications and 771 Chinese publications were included for bibliometric analyses. The overall output of English publications was relatively small from 2005 to 2024, the annual average publication was 7.90 publications. Parasites & Vectors was the most productive journal by the number of publications (21 publications). The three most productive authors included Li Shizhu (24 publications), Zhou Xiaonong (13 publications), and Yang Kun (12 publications), and the three most productive institutions included Chinese Center for Disease Control and Prevention (49 publications), the WHO (27 publications), and Fudan University (25 publications). The annual average number of Chinese publications was high from 2005 to 2015 (57.73 publications), and reduced to 15.11 publications during the period from 2016 to 2024, with Chinese Journal of Schistosomiasis Control as the most productive journal (241 publications). The three most productive authors included Wang Wanxian (18 publications), Sun Qixiang (16 publications), and Dai Jianrong (16 publications), and the three most productive institutions included Jiangsu Institute of Parasitic Diseases (55 publications), Chinese Center for Disease Control and Prevention (47 publications), and Hubei Uni-versity (38 publications). Among the 158 English publications, molluscicidal effect, climate change, geographic information, biological control, machine learning were current research hotspots, and the Yangtze River and elimination were emerging research hotspots. Among the 771 Chinese publications, molluscicidal effect, niclosamide, comprehensive management, molluscicide, effectiveness evaluation, marshland, and endophyte were current research hotspots, and the future research hotspots shifted to molluscicidal effect and pyriclobenzuron. Limited attention is paid to the research on O. hupensis control across the world. The Yangtze River, elimination, molluscicidal effect, and pyriclobenzuron may be future research hotspots. High attention is recommended to be paid to the research on O. hupensis control, and development of diverse approaches for O. hupensis control is of urgent needs. We should continue to attach importance to the control research of O. hupensis and strengthen the exploration of diverse snail extermination and control methods. ［摘要］ 目的 对2005—2024年发表的钉螺控制相关研究中英文文献进行分析, 了解该领域的研究现状和热点。方法 分别在Web of Science核心合集数据库和中国期刊全文数据库中检索2005—2024年发表的有关钉螺控制研究的英 文和中文文献, 使用CiteSpace 6.3.1软件分析各年发文情况, 并构建作者、机构合作网络图谱; 提取文献关键词绘制关键 词共现、突现和时间线图谱, 分析钉螺控制研究热点。结果 共纳入158篇英文文献和771篇中文文献进行文献计量学 分析。2005—2024年英文文献发文量总体较少, 年均发文量为7.90篇; 《Parasites & Vectors》是刊载文献最多的期刊 (21 篇); 发文量居前3位的作者分别为Li Shizhu (24篇)、Zhou Xiaonong (13篇)、Yang Kun (12篇), 居前3位的机构分别是中 国疾病预防控制中心 (49篇)、WHO (27篇)、复旦大学 (25篇)。2005—2015年中文文献年均发文量为57.73篇, 2016— 2024年降至15.11篇; 《中国血吸虫病防治杂志》是刊载文献最多的期刊 (241篇); 发文量居前3位的作者分别为王万贤 (18篇)、孙启祥 (16篇)、戴建荣 (16篇), 排名前3位的机构分别是江苏省血吸虫病防治研究所 (55篇)、中国疾病预防控 制中心 (47篇)、湖北大学 (38篇)。158篇英文文献中, 灭螺效果 (molluscicidal activity)、气候变化 (climate change)、地理 信息 (geographic information)、生物控制 (biological control)、机器学习 (machine learning) 是目前的研究热点, 长江 (Yangtze River) 和消除 (elimination) 是新的研究热点。771篇中文文献中, 灭螺效果、氯硝柳胺、综合治理、灭螺剂、效果评价、江 滩、内生真菌是目前的研究热点, 未来研究热点转向灭螺效果和吡螺脲。结论 国内外对钉螺控制研究的关注度较低, 长江、消除、灭螺效果、吡螺脲可能是今后的研究热点, 应继续重视钉螺控制研究, 加强对多样化灭螺、控螺方式的探讨。.
To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate. ［摘要］ 目的 克隆、表达日本血吸虫 (Schistosoma japonicum, Sj) 热休克同源蛋白20 (heat shock cognate protein 20, Hsc20) (SjHsc20), 并对其生物学特性进行初步研究。方法 采用PCR法扩增SjHsc20基因目的片段, 构建重组表达载体 pET-28a (+)-SjHsc20, 并将其转化入大肠埃希菌BL21 (DE3)。采用异丙基-β-D-硫代半乳糖苷 (isopropyl β-D-thiogalactopyranoside, IPTG) 诱导表达并纯化重组SjHsc20 (rSjHsc20) 蛋白, 采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳 (sodium dodecyl sulfate-polyacrylamide gel electrophoresis, SDS-PAGE) 验证重组蛋白表达, 采用Western blotting检测重组蛋白免疫原 性。采用实时荧光定量PCR (real-time quantitative reverse transcription polymerase chain reaction, RT-qPCR) 法检测SjH-sc20 基因在不同发育期虫体及不同性别成虫中的转录水平。30只雌性6～8周龄BALB/c小鼠随机分为rSjHsc20免疫组、PBS对照组、ISA 206佐剂组, 每组10只。在实验第1、15、31天, rSjHsc20免疫组、PBS对照组、ISA 206佐剂组小鼠分别皮 下免疫注射20 μg rSjHsc20重组蛋白及等体积PBS和ISA 206佐剂, 最后1次免疫14 d后, 各组小鼠经腹部贴片法感染日 本血吸虫尾蚴 (40 ± 2) 条。采用间接酶联免疫吸附试验 (enzyme-linked immunosorbent assay, ELISA) 检测各组小鼠血清 抗rSjHsc20特异性IgG及其亚型IgG1、IgG2a抗体水平, 以及血清中抗rSjHsc20抗体滴度, 并计算IgG1/IgG2a比值。感染 42 d后, 剖杀各组小鼠, 采用肝门静脉灌注法收集虫体并计数, 计算每克肝脏虫卵数 (eggs per gram, EPG) 以及rSjHsc20免 疫组相对于PBS对照组的减虫率和减卵率, 以评估rSjHsc20蛋白免疫保护效果。结果 SjHsc20基因开放阅读框长 756 bp、编码252个氨基酸, rSjHsc20蛋白相对分子质量约为29 kDa。rSjHsc20可被感染日本血吸虫的小鼠血清以及免疫 该蛋白后获得的小鼠血清识别。7、12、21、28、35、42 d虫体SjHsc20 基因相对转录水平分别为1.001 4 ± 0.065 7、2.268 3 ± 0.129 2、1.378 5 ± 0.160 4、1.196 4 ± 0.244 0、1.646 3 ± 0.226 1、1.758 0 ± 0.611 1, 差异有统计学意义 (F = 38.45, P < 0.000 1), 其中12 d虫体内SjHsc20 基因转录水平高于其他时期虫体 (P 均 < 0.05)。第3次免疫后, PBS对照组、ISA 206佐剂组和 rSjHsc20免疫组小鼠血清IgG抗体水平分别为0.106 6 ± 0.010 7、0.108 3 ± 0.010 4、0.553 2 ± 0.069 1, IgG1抗体水平分别为 0.137 3 ± 0.054 0、0.181 1 ± 0.096 8、1.765 8 ± 0.221 1, IgG2a抗体水平分别为0.280 3 ± 0.197 6、0.274 0 ± 0.146 3、1.560 4 ± 0.106 0, 差异均有统计学意义 (F = 397.70、401.00和229.70, P 均< 0.000 1); 两两比较结果显示, rSjHsc20免疫组小鼠血清 IgG、IgG1和IgG2特异性抗体水平均高于其他两组 (P均< 0.000 1)。3组小鼠血清IgG1/IgG2a比值分别为0.428 4 ± 0.199 8、0.559 9 ± 0.181 1、1.177 2 ± 0.143 6, 差异有统计学意义 (F = 43.97, P < 0.000 1); rSjHsc20免疫组IgG1/IgG2a比值> 1, 且高 于PBS对照组和ISA 206佐剂组 (P 均< 0.001)。3次免疫后, rSjHsc20免疫组小鼠血清抗体滴度均在1∶16 384 以上, 表明 rSjHsc20具有较强免疫原性。在尾蚴攻击42 d后, PBS对照组、ISA 206佐剂组、rSjHsc20免疫组小鼠平均虫荷数分别为 (16.60 ± 5.75)、(15.80 ± 5.58)、(14.40 ± 5.75) 条/鼠, 差异无统计学意义 (F = 0.50, P > 0.05); 3组小鼠EPG分别为68 370 ± 22 690、67 972 ± 19 502、41 075 ± 13 251, 差异有统计学意义 (F = 4.55, P < 0.05); 两两比较结果显示, PBS对照组和ISA 206佐剂组EPG均低于rSjHsc20免疫组 (P 均< 0.05)。rSjHsc20免疫组小鼠减虫率和减卵率分别为13.25%和39.92%。结论 成功克隆、表达了SjHsc20基因, rSjHsc20蛋白可诱导小鼠产生一定的免疫保护效果。上述结果为后续研究SjH-sc20生物学功能及评估其作为疫苗候选分子的潜能奠定了基础。.
To construct an index system for assessment of schistosomiasis transmission risk following natural disasters such as rainstorms, floods, earthquakes, mudslides, and landslides, so as to provide insights into rapid identification of schistosomiasis transmission risk post-disasters and formulation of targeted schistosomiasis control strategies. An initial framework for the index system for assessment of schistosomiasis transmission risk following natural disasters was drafted through literature review, brainstorming, and focus group discussions. Two rounds of expert correspondence consultations were conducted using the Delphi method to refine and finalize the system, and the degrees of expert activeness, authority and endorse ment, and consensus were evaluated. In addition, the weights of each index were calculated using the analytic hierarchy process. A total of 18 experts participated in the consultation. The expert positive coefficients were 100.00% and 94.44% for two rounds of consultations, with authority coefficients of 0.92 and 0.94, respectively. The coefficients of coordination on the index importance, rationality and operability were 0.209, 0.185, 0.222 and 0.407, 0.214, 0.257 for two rounds of consultations, respectively, and all consistency tests were statistically significant (χ2 = 246.771 to 505.278, all P values < 0.001). Following two rounds of expert consultations, an index system consisting of 6 first-level indicators, 15 second-level indicators, and 49 third-level indicators was ultimately constructed. In terms of first-level indicators, "disaster situation", "previous epidemics", "healthcare guarantee", "response capacity" and "emergency recovery" had the highest weights, each at 18.18%. Regarding second-level indicators, "Schistosoma japonicum infections in animals", "S. japonicum infections in snails" and "medical treatment" had the highest weights, each at 7.35%. In terms of third-level indicators, ten items had the highest weights, including "identification of schistosomiasis cases", "detection of S. japonicum infections in wild feces", "detection of S. japonicum infections in snails", "reserves of schistosomiasis diagnostic/testing reagents and consumables", "reserves of chemotherapy agents for human and animal schistosomiasis", "reserves of cercariacides", "periodical surveillance on schistosomiasis", "identification of schistosomiasis transmission risk and timely response", "normal provision of diagnosis and treatment services" and "post-disaster schistosomiasis surveillance", each at 2.40%. A scientific, systematic, and practical index system has been constructed for assessment of schistosomiasis transmission risk following natural disasters, which may provide insights into rapid post-disaster identification of schistosomiasis transmission risk, formulation of targeted schistosomiasis control strategies and optimization of resource allocation. ［摘要］ 目的 构建暴雨、洪涝、地震、泥石流、滑坡等自然灾害后血吸虫病传播风险评估指标体系, 为灾后快速识别传 播风险、制定针对性防控策略提供科学依据。方法 通过文献检索、头脑风暴法和专题小组讨论, 初步拟定自然灾害后 血吸虫病传播风险评估指标体系框架。采用德尔菲法, 通过函询的方式进行两轮专家咨询。根据专家咨询结果, 对指标 体系进行调整, 并评价专家积极程度、专家权威程度、专家认可程度及专家意见一致性, 采用层次分析法计算各指标权 重。结果 共有18名专家参与咨询。两轮咨询的专家积极系数分别为100.00%和94.44%, 权威系数分别为0.92和 0.94, 指标重要性、合理性和可操作性评分的协调系数分别为0.209、0.185、0.222和0.407、0.214、0.257, 一致性检验均有 统计学意义 (χ2 = 246.771～505.278, P 均< 0.001)。经过两轮专家咨询, 最终形成包括6个一级指标、15个二级指标和49 个三级指标的评估体系。一级指标中, “受灾情况”、“既往疫情”、“卫生保障”、“响应能力”和“应急恢复”权重最高, 均为 18.18%。二级指标中, “动物感染”、“钉螺感染”、“医疗救治”权重最高, 均为7.35%。三级指标中, “是否发现病人”、“是 否检出阳性野粪”、“是否检出感染性钉螺”、“是否储备血吸虫病诊断、检测试剂及耗材”、“是否储备人畜血吸虫病治疗药 物”、“是否储备灭蚴药物”、“是否定期开展血吸虫病监测”、“是否发现风险并及时处置”、“是否可以正常提供诊疗服务” 和“灾后是否开展血吸虫病监测”10项指标权重最高, 均为2.40%。结论 本研究构建的自然灾害后血吸虫病传播风险 评估指标体系具有科学性、系统性和可操作性, 可为灾后快速识别血吸虫病传播风险、制定针对性防控策略及优化资源 配置提供科学参考。.
To evaluate the value of inquiry examinations for diagnosis of taeniasis in field investigations, so as to provide insights into improving the efficiency and accuracy of taeniasis control programmes. Four taeniasis-endemic villages were sampled in Dali Bai Autonomous Prefecture, Yunnan Province in October 2023 as survey sites, and at least 305 permanent residents at ages of over 3 years were randomly sampled from each site. Face-to-face inquiries were performed with structured questionnaires to investigate participants' history and frequency of proglottids release during the past year, consumption of raw or undercooked meat products or pork liver during the past six months, history and time of deworming. Participants' stool samples were collected, and Taenia eggs were detected in stool samples using Kato-Katz technique (two slides of one stool sample). Egg-positive individuals or participants with a history of proglottids release during the past year were given diagnostic deworming with areca nuts and pumpkin seeds. The detection of Taenia eggs using Kato-Katz technique and release of Taenia worms or proglottids following diagnostic deworming served as a gold standard to evaluate the diagnostic efficiency of inquiry examinations for taeniasis. The receiver operating characteristic (ROC) curves of inquiry examinations for diagnosis of taeniasis were plotted, and the area under the curve (AUC) was calculated. In addition, Taenia worms or proglottids released following diagnostic deworming were subjected to multiplex PCR assay and Sanger sequencing for species identification. A total of 1 842 participants were included, and 1 842 valid questionnaires were recovered. A total of 1 533 stool samples were collected, among which 25 egg-positives were identified. Questionnaire surveys showed that 1 305 respondents had consumed raw or undercooked meat products during the past six months, and 42 respondents had a history of proglottids release during the past year. Diagnostic deworming was given to the aforementioned 25 egg-positive individuals and 42 participants who self-reported a history of proglottids release during the past year, and 33 participants had a release of complete worms. Following exclusion of records of individuals receiving deworming during the past 3 months, during the past year, and all participants with a history of deworming, the AUCs were 0.767 (Z = 4.71, P < 0.001), 0.762 (Z = 4.51, P < 0.001), and 0.762 (Z = 4.52, P < 0.001) for diagnosis of Taenia infections with "self-reported history of proglottids release during the past year", respectively, and pairwise comparisons showed no statistically significant differences (D values, -0.01 to 0.33; all P values > 0.05). Following exclusion of records of individuals receiving deworming during the past 3 months, during the past year, and all participants with a history of deworming, the AUCs were 0.797 (Z = 4.71, P < 0.001), 0.835 (Z = 4.17, P < 0.001), and 0.847 (Z = 3.98, P < 0.001) for diagnosis of Taenia infections with "self-reported history of twice proglottids releases", respectively, and pairwise comparisons showed no statistically significant differences (D values, -0.43 to -0.10; all P values > 0.05). Following exclusion of records of all individuals receiving deworming, the diagnostic sensitivity and specificity of "self - reported history of twice proglottids releases" were 87.50% and 81.82%, respectively. Following exclusion of records of individuals receiving deworming at different intervals, the AUCs were 0.529 to 0.532 for diagnosis of Taenia infections with "self-reported consumption of raw or undercooked meat products or pork liver during the past 6 months" (all P values > 0.05). Among 33 individuals with releases of complete Taenia worms, 31 individuals were identified with T. asiatica infections (93.94%), with a mean worm burden of (1.39 ± 0.72) worms/person, and 2 were identified with T. saginata infections (6.06%), with one worm in each participant. A history of Taenia proglottids release during the past year as revealed by inquiry examinations exhibit a diagnostic value for taeniasis and may serve as an initial screening tool for field taeniasis screening. Increasing the frequency of Taenia proglottids release during inquiry examinations and exclusion of records of individuals receiving deworming during the analysis may improve the value for diagnosis of taeniasis. ［摘要］目的 评价现场调查中采用询检法诊断带绦虫病的价值, 为进一步提升带绦虫病防控工作效率和精准度提供科学依据。方法2023年10月, 在云南省大理白族自治州选择有带绦虫病流行的4个村作为调查点, 各调查点分别随机抽取3周岁以上常住居民开展调查, 调查人数均≥ 305例。采用结构化问卷以面对面方式对调查对象开展询检, 了解其近1年内排节片史和排节片次数, 近半年内食用生肉及其制品、生猪肝等未熟肉制品和是否服驱虫药物及服用驱虫药时间等信息。收集调查对象粪便样本, 采用改良加藤厚涂片法 (1粪2检) 检测粪便中带绦虫虫卵。对粪检阳性或自述近1年内有排节片史者采用槟榔-南瓜籽法进行诊断性驱虫治疗。以改良加藤厚涂片法检出带绦虫虫卵及诊断性驱虫治疗驱出虫体或节片作为“金标准”, 评价询检法用于带绦虫病的诊断效能, 绘制受试者工作特征 (receiver operating characteristic, ROC) 曲线并计算曲线下面积。此外, 收集调查对象在诊断性驱虫治疗中排出的虫体或节片, 采用多重PCR法及Sanger测序技术鉴定带绦虫虫种。结果共纳入1 842例调查对象, 收集1 842份有效问卷; 共采集1 533份粪便样本, 其中带绦虫虫卵阳性样本25份。问卷调查结果表明, 自述近半年内食用生肉及其制品、生猪肝等未熟肉制品者1 305例, 近1年内有排节片史者42例。对上述25例粪检阳性者及42例自述近1年内有排节片史者进行诊断性驱虫治疗, 其中33例驱出完整虫体。排除近3个月内、近1年内有服药史者及全部有服药史者记录后, “自述近1年内有排节片史”诊断带绦虫病的ROC曲线下面积分别为0.767 (Z = 4.71, P < 0.001) 、0.762 (Z = 4.51, P < 0.001) 、0.762 (Z = 4.52, P < 0.001), 两两比较显示差异均无统计学意义 (D = –0.01 ~ 0.33, P 均> 0.05) 。排除近3个月内、近1年内有服药史者及全部有服药史者记录后, “自述排节片次数为2次”诊断带绦虫病的ROC曲线下面积分别为0.797 (Z = 4.71, P < 0.001) 、0.835 (Z = 4.17, P < 0.001) 、0.847 (Z = 3.98, P < 0.001), 两两比较显示差异均无统计学意义 (D = –0.43 ~ –0.10, P 均> 0.05); 排除所有有服药史者记录后, “自述排节片次数为2次”的诊断灵敏度和特异度分别为87.50%和81.82%。在排除不同服药间隔者记录后, “自述近半年内食用生肉及其制品、生猪肝等未熟肉制品”诊断带绦虫病的ROC曲线下面积在0.529 ~ 0.532, 但差异均无统计学意义 (P 均> 0.05) 。经虫种鉴定, 驱出虫体的33例调查对象中, 31 例 (93.94%) 感染亚洲带绦虫, 平均感染度为 (1.39 ± 0.72) 条/例; 2例 (6.06%) 感染牛带绦虫, 感染度均为1条/例。结论询检法对于带绦虫病诊断具有一定价值, 可作为带绦虫病现场筛查的初筛工具。询检时进一步增加排节片次数等信息以及在分析过程中排除有服药史记录, 均可提高诊断价值。.
To evaluate the effectiveness of schistosomiasis surveillance in Jiangsu Province during the stage moving from transmission control to transmission interruption, and to analyze the current risk and challenges, so as to provide the evidence for achieving the target of schistosomiasis elimination. Schistosomiasis surveillance data were collected from Jiangsu Province from 2012 to 2024, and the endemic areas, Schistosoma japonicum infections in humans and livestock, Oncomelania hupensis snail distribution and implementation of integrated interventions were descriptively analyzed. In addition, the trends in areas with snails, seroprevalence of human S. japonicum infections and numbers of advanced schistosomiasis cases were assessed using a Joinpoint regression model. The endemic areas of schistosomiasis continued to shrink in Jiangsu Province from 2012 to 2024, with the number of schistosomiasis-eliminated counties (cities, districts) increasing from 53 (75.71%) to 63 (96.92%), and interruption of schistosomiasis transmission was achieved across the province. A total of 4 600 300 person-times were tested for serum antibodies against S. japonicum, with 28 719 person-times positive detected; and 616 500 person-times were tested S. japonicum infections among local residents in Jiangsu Province from 2012 to 2024, with only 3 egg-positives detected, and no egg-positives found since 2017. A total of 187 600 herd-times were tested for schistosomiasis in livestock, and no S. japonicum infections were found. O. hupensis snail survey was performed covering 1 018 408.97 hm2, and a total of 35 556.35 hm2 was found with snail-infested habitats, including 174.40 hm2 of emerging snail-infested habitats. A total of 1 102 800 O. hupensis snails were identified for S. japonicum infections, and no infections were found. The areas of snail-infested habitats appeared a tendency towards a rise in Jiangsu Province from 2019 to 2023 (APC = 23.67%, P < 0.05), and the actual areas of snail-infested habitats appeared a tendency towards a decline from 2012 to 2015 (APC = -22.77%, P < 0.05), and towards a rise from 2015 to 2023 (APC = 9.76%, P < 0.01). The seroprevalence of anti-S. japonicum antibodies appeared a tendency towards a decline among residents in Jiangsu Province from 2017 to 2023 (APC = -14.92%, P < 0.01). In addition, the number of newly diagnosed advanced schistosomiasis cases appeared a tendency towards a decline from 2012 to 2024 (APC = -12.02%, P < 0.01), and the numbers of advanced schistosomiasis patients requiring treatment showed a tendency towards a decline from 2012 to 2021 (APC = -10.56%, P < 0.01) and from 2021 to 2023 (APC = -20.06%, P < 0.01). Great progresses had been achieved in schistosomiasis control in Jiangsu Province following transmission control, and transmission interruption had been achieved; however, there are still snail-infested habitats. High-intensity surveillance and integrated control are required to be maintained to advance the achievement of the target of schistosomiasis elimination in Jiangsu Province. ［摘要］ 目的 分析2012—2024年江苏省血吸虫病监测结果, 评价江苏省达到血吸虫病传播控制标准后向传播阻断标 准推进的防治成效, 分析当前存在的风险与挑战, 为实现消除血吸虫病目标提供科学依据。方法 收集2012—2024年 江苏省血吸虫病监测数据, 对流行范围、人群和家畜感染情况、钉螺分布及综合防控措施等指标进行描述性分析。采用 Joinpoint回归模型对血吸虫病疫情指标逐年变化趋势进行建模分析。结果 2012—2024年, 江苏省血吸虫病流行区范 围持续压缩, 达到血吸虫病消除标准的县 (市、区) 数从53个 (占75.71%) 增至63个 (占96.92%), 全省均达到血吸虫病传 播阻断标准。2012—2024年, 江苏省累计开展本地人群血清抗血吸虫抗体检查460.03万人·次, 检出阳性28 719人·次; 人群病原学检测61.65万人·次, 检出阳性3例, 自2017年起未查出粪检阳性病人; 累计开展家畜血吸虫病查病18.76万 头·次, 未查出血吸虫感染阳性家畜; 累计开展钉螺分布调查1 018 408.97 hm2, 累计查出有螺面积35 556.35 hm2, 其中新 发现有螺面积174.40 hm2; 解剖钉螺110.28万只, 未检出血吸虫感染阳性钉螺。2012—2024年, 江苏省查出有螺面积于 2019—2023年呈上升趋势 (APC = 23.67%, P < 0.05); 实有钉螺面积于2012—2015年呈下降趋势 (APC = −22.77%, P < 0.05), 于2015—2023年呈上升趋势 (APC = 9.76%, P < 0.01); 人群血清抗血吸虫抗体阳性率于2017—2023年呈下降趋势 (APC = −14.92%, P < 0.01); 新发现晚期血吸虫病 (晚血) 病人数呈下降趋势 (APC = −12.02%, P < 0.01), 2012—2021年 (APC = −10.56%, P < 0.01) 和2021—2023年 (APC = −20.06%, P < 0.01) 需治疗晚血病人数均呈下降趋势。结论 江苏省 在达到血吸虫病传播控制标准后, 防治工作进展显著, 基本达到传播阻断标准; 但有螺环境依然存在, 应保持高强度监测 和综合防控, 推动血吸虫病消除目标的实现。.
To investigate the suitable habitats of Phlebotomus chinensis in Gansu Province, so as provide insights into effective management of mountain-type zoonotic visceral leishmaniasis (MT-ZVL). The geographical coordinates of locations where MT-ZVL cases were reported were retrieved in Gansu Province from 2015 to 2023, and data pertaining to 26 environmental variables were captured, including 19 climatic variables (annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the coldest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter), five geographical variables (elevation, annual normalized difference vegetation index, vegetation type, landform type and land use type), and two population and economic variables (population distribution and gross domestic product). Twelve species distribution models were built using the biomod2 package in R project, including surface range envelope (SRE) model, generalized linear model (GLM), generalized additive model (GAM), multivariate adaptive regression splines (MARS) model, generalized boosted model (GBM), classification tree analysis (CTA) model, flexible discriminant analysis (FDA) model, maximum entropy (MaxEnt) model, optimized maximum entropy (MAXNET) model, artificial neural network (ANN) model, random forest (RF) model, and extreme gradient boosting (XGBOOST) model. The performance of 12 models was evaluated using the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS), and Kappa coefficient, and single models with high performance was selected to build the optimal ensemble models. Factors affecting the survival of Ph. chinensis were identified based on climatic, geographical, population and economic variables. In addition, the suitable distribution areas of Ph. chinensis were predicted in Gansu Province under shared socioeconomic pathway 126 (SSP126), SSP370 and SSP585 scenarios based on climatic data during the period from 1991 to 2020, from 2041 to 2060 (2050s), and from 2081 to 2100 (2090s) . A total of 11 species distribution models were successfully built for prediction of potential distribution areas of Ph. chinensis in Gansu Province, and the RF model had the highest predictive accuracy (AUC = 0.998). The ensemble model built based on the RF model, XGBOOST model, GLM, and MARS model had an increased predictive accuracy (AUC = 0.999) relative to single models. Among the 26 environmental factors, precipitation of the wettest quarter (12.00%), maximum temperature of the warmest month (11.58%), and annual normalized difference vegetation index (11.29%) had the greatest contributions to suitable habitats distribution of Ph. sinensis. Under the climatic conditions from 1991 to 2020, the potential suitable habitat area for Ph. chinensis in Gansu Province was approximately 5.80 × 104 km2, of which the highly suitable area was 1.42 × 104 km2, and primarily concentrated in the southernmost region of Gansu Province. By the 2050s, the unsuitable and lowly suitable areas for Ph. chinensis in Gansu Province had decreased by varying degrees compared to that of 1991 to 2020 period, while the moderately and highly suitable areas exhibited expansion and migration. By the 2090s, under the SSP126 scenario, the suitable habitat area for Ph. chinensis increased significantly, and under the SSP585 scenario, the highly suitable areas transformed into extremely suitable areas, also showing substantial growth. Future global warming is conducive to the survival and reproduction of Ph. chinensis. From the 2050s to the 2090s, the highly suitable areas for Ph. chinensis in Gansu Province will be projected to expand northward. Under the SSP126 scenario, the suitable habitat area for Ph. chinensis in Gansu Province is expected to increase by 194.75% and 204.79% in the 2050s and 2090s, respectively, compared to that of the 1991 to 2020 period. Under the SSP370 scenario, the moderately and highly suitable areas will be projected to increase by 164.40% and 209.03% in the 2050s and 2090s, respectively, while under the SSP585 scenario, they are expected to increase by 195.98% and 211.66%, respectively. The distribution of potential suitable habitats of Ph. sinensis gradually shifts with climatic changes. Intensified surveillance and management of Ph. sinensis is recommended in central and eastern parts of Gansu Province to support early warning of MT-ZVL. ［摘要］ 目的 识别甘肃省中华白蛉适生区, 为有效防控山丘型内脏利什曼病 (mountain-type zoonotic visceral leishmaniasis, MT-ZVL) 提供参考。方法 收集 2015—2023 年甘肃省MT-ZVL报告病例信息, 获取病例分布点地理坐标。收集 26 个环境变量数据, 包括年平均温度、平均气温日较差、等温性、气温季节性变化标准差、最暖月最高气温、最冷月最低气 温、温度年较差、最湿季度平均气温、最干季度平均气温、最暖季度平均气温、最冷季度平均气温、年降水量、最湿月降水 量、最干月降水量、降水量季节性变化标准差、最湿季度降水量、最干季度降水量、最暖季度降水量、最冷季度降水量等 19 个气候变量, 海拔、年度归一化植被指数、植被类型、地形地貌、土地利用类型等 5 个地理变量和国内生产总值、人口分 布 2 个人口经济变量。采用 R 软件Biomod2 软件包建立 12 个物种分布模型, 包括表面分布区分室模型 (surface range envelope, SRE)、广义线性模型 (generalized linear model, GLM)、广义加性模型 (generalized additive model, GAM)、多元适应 回归样条 (multivariate adaptive regression splines, MARS)、推广回归树 (generalized boosted model, GBM)、分类树分析 (classification tree analysis, CTA)、柔性判别分析 (flexible discriminant analysis, FDA)、最大熵模型 (maximum entropy model, MAXENT)、优化版最大熵 (optimized maximum entropy model, MAXNET)、人工神经网络 (artificial neural network, ANN)、随 机森林 (random forest, RF) 和极限梯度提升 (extreme gradient boosting, XGBOOST)。利用受试者工作特征曲线下面积 (area under curve, AUC)、真实技巧统计量 (true skill statistics, TSS) 和 Kappa 系数对各模型进行性能评价, 选择性能较好的单一模型构建最优组合模型, 基于气候、地形和社会经济因子分析影响中华白蛉生存的因素。根据 1991—2020 年和 2041—2060 年 (2050s)、2081—2100 年 (2090s) 气候数据, 分别对共享社会经济路径 (shared socioeconomic pathways, SSP) 126、SSP370 和 SSP585 气候情景下甘肃省中华白蛉适宜分布区进行预测。结果 成功建立了 11 种预测甘肃省中华白蛉潜在 分布区的模型。其中随机森林模型预测准确度最高 (AUC = 0.998)。以RF、XGBOOST、GLM 和 MARS 4 种表现较好的单 一模型构建的组合模型 (AUC = 0.999) 较各单一模型准确度有所提升。26 个环境变量中, 最湿季度降水量、最暖月最高 气温和年度归一化植被指数对中华白蛉适宜生境分布影响最大, 贡献率分别为 12.00%、11.58% 和 11.29%。1991—2020 年气候条件下, 甘肃省中华白蛉潜在适生区面积约 5.80 × 104 km2, 其中高适生区为 1.42 × 104 km2, 主要集中于陇南南部。2050s 甘肃省中华白蛉非适生区与低适生区面积较 1991—2020 年均不同程度降低, 但中、高适生区面积存在扩张迁移趋 势; 在 SSP126 情景下 2090s 中华白蛉适生区面积将大幅增加, 在 SSP585 情景下 2090s 中华白蛉高适生区转变为极高适生 区的面积将大幅增加。未来全球气候变暖有利于中华白蛉的生存繁衍, 2050s 至 2090s 甘肃省中华白蛉高适生区将呈向 北扩展的趋势。在SSP126情景下, 2050s 与 2090s 甘肃省中华白蛉适生区面积分别较 1991—2020 年增加 194.75% 和 204.79%; 在SSP370 情景下, 2050s 与 2090s 甘肃省中华白蛉中、高适生区面积较 1991—2020 年分别增加 164.40% 和 209.03%, 在SSP585情景下分别增加 195.98% 和 211.66%。结论 甘肃省中华白蛉潜在适生区范围会随气候变化而逐渐 迁移, 未来可有针对性地在陇中和陇东地区对中华白蛉加强监测和防控, 从而为 MT-ZVL 早期预警提供支持。.
To investigate the prevalence of Schistosoma japonicum infections in wild rodents in schistosomiasis-endemic areas of China, so as to provide insights into formulation of technical guidelines for monitoring of and the precise control strategy for S. japonicum infections in wild rodents during the elimination phase. Two administrative villages where schistosomiasis was historically highly prevalent were selected each from Dongzhi County, Anhui Province, and Duchang County, Jiangxi Province as study villages. Wild rodents were captured from study villages with baited traps or cages at night in June and September, 2021. The number of rodents captured was recorded, and the rodent species was characterized based on morphologi-cal characteristics. Liver tissues were sampled from captured rodents for macroscopical observation of the presence of egg granulomas, and S. japonicum infection was detected simultaneously using liver tissue homogenate microscopy, examinations of mesenteric tissues for parasites, and modified Kato-Katz thick smear technique (Kato-Katz technique). A positive S. japonicum infection was defined as detection of S. japonicum eggs or adult worms by any of these methods. The rate of wild rodent capture and prevalence of S. japonicum infections in wild rodents were compared in different study villages and at different time periods, and the detection of S. japonicum infections in wild rodents was compared by different assays. The overall rate of wild rodent capture was 8.28% (237/2 861) in Dongzhi County, and the wild rodent capture rates were 9.24% (133/1 439) and 7.31% (104/1 422) in two study villages (χ2 = 3.503, P = 0.061), and were 8.59% (121/1 409) and 7.99% (116/1 452) in June and September, 2021, respectively (χ2 = 0.337, P = 0.561). The overall rate of wild rodent capture was 3.72% (77/2 072) in Duchang County, and the wild rodent capture rates were 6.91% (67/970) and 0.91% (10/1 102) in two study villages (χ2 = 51.901, P < 0.001), and were 4.13% (39/945) and 3.37% (38/1 127) in June and September, 2021, respectively (χ2 = 0.815, P = 0.365). Rattus norvegicus was the predominant rodent species captured in both counties, accounting for 70.04% (166/237) of all captured wild rodents in Dongzhi County and 88.31% (68/77) in Duchang County. No S. japonicum infection was detected in wild rodents captured in Duchang County. Nevertheless, the overall prevalence of S. japonicum infections was 51.05% (121/237) in wild rodents captured in Dongzhi County, with prevalence rates of 50.38% (67/133) and 51.92% (54/104) in two study villages (χ2 = 0.098, P = 0.755), and 54.31% (63/116) and 47.93% (58/121) in September and June, 2021, respectively (χ2 = 0.964, P = 0.326). Of 237 wild rodents captured in Dongzhi County, there were 140 (59.07%) rodents with visible hepatic egg granulomas, 117 (49.47%) tested positive for S. japonicum eggs by liver tissue homogenate microscopy, 34 (14.35%) tested positive for S. japonicum eggs with Kato-Katz technique; however, no adult S. japonicum worms were detected in mesenteric tissues. In addition, hepatic egg granulomas were found in all wild rodents tested positive for S. japonicum eggs with liver tissue homogenate microscopy. The rate of wild rodent capture and prevalence of S. japonicum infection in wild rodents vary greatly in schistosomiasis-endemic areas of China, and the prevalence of S. japonicum infection is slightly higher in wild rodents captured in autumn than in summer. Liver tissue is recommended as the preferred sample for surveillance of S. japonicum infection in wild rodents, and a combination of macroscopical observation of hepatic egg granulomas and liver tissue homogenate microscopy may be a standard method for surveillance of S. japonicum infection in wild rodents. ［摘要］ 目的 了解我国血吸虫病流行区野鼠血吸虫感染率, 为制定消除阶段野鼠血吸虫感染监测技术规范以及精准 防控策略提供依据。方法 在安徽省东至县和江西省都昌县各选择2个历史血吸虫病疫情较重的行政村作为调查村, 分别于2021年6月和9月采用夹 (笼) 夜法捕捉野鼠, 统计野鼠捕获数量, 采用形态学方法鉴定鼠种。捕获的野鼠剖杀 后, 取野鼠肝脏, 肉眼观察其表面是否有虫卵结节; 随后采用肝脏组织匀浆镜检法、肠系膜组织血吸虫虫体检查法和改良 加藤厚涂片 (Kato-Katz) 法对野鼠血吸虫感染情况进行平行检测, 任一方法检出血吸虫虫卵或虫体则判定为血吸虫感染 阳性。比较不同调查村、调查时间野鼠捕获率、血吸虫感染率及不同方法对野鼠血吸虫感染检出率的差异。结果 安徽 省东至县野鼠捕获率为8.28% (237/2 861); 2个调查村野鼠捕获率分别为9.24% (133/1 439) 和7.31% (104/1 422), 差异无 统计学意义 (χ2 = 3.503, P = 0.061); 2021年6月和9月野鼠捕获率分别为8.59% (121/1 409) 和7.99% (116/1 452), 差异无 统计学意义 (χ2 = 0.337, P = 0.561) 。江西省都昌县野鼠捕获率为3.72% (77/2 072); 2个调查村野鼠捕获率分别为6.91% (67/970) 和0.91% (10/1 102), 差异有统计学意义 (χ2 = 51.901, P < 0.001); 2021年6月和9月野鼠捕获率分别为4.13% (39/ 945) 和3.37% (38/1 127), 差异无统计学意义 (χ2 = 0.815, P = 0.365)。两县捕获野鼠均以褐家鼠为主, 其中东至县捕获褐 家鼠占捕获野鼠总数的70.04% (166/237) 、都昌县占88.31% (68/77) 。都昌县捕获野鼠未发现血吸虫感染阳性野鼠。东 至县捕获野鼠的血吸虫感染率为51.05% (121/237); 2个调查村野鼠血吸虫感染率分别为50.38% (67/133) 和51.92% (54/ 104), 差异无统计学意义 (χ2 = 0.098, P = 0.755); 2021 年9 月捕获的野鼠血吸虫感染率 (54.31%, 63/116) 略高于6 月 (47.93%, 58/121), 差异无统计学意义 (χ2 = 0.964, P = 0.326) 。此外, 在东至县捕获的237只野鼠中, 140只 (59.07%) 发现 肝脏虫卵结节、117只 (49.47%) 肝脏组织匀浆镜检发现血吸虫虫卵、34只 (14.35%) 粪便Kato-Katz法检出血吸虫虫卵, 但 肠系膜组织检查均未发现血吸虫虫体; 所有肝脏组织匀浆镜检检出血吸虫虫卵的野鼠均发现存在肝脏虫卵结节。结论 我国不同血吸虫病流行区野鼠捕获率及野鼠血吸虫感染率差异较大, 秋季捕获的野鼠血吸虫感染率略高于夏季。肝 脏样本可作为野鼠血吸虫感染监测的首选样本, 肝脏虫卵结节观察法结合肝脏组织匀浆镜检法可作为监测的标准方法。.
To investigate the epidemiological characteristics of cross-county imported dengue fever cases within Yunnan province in 2023, so as to provide insights into formulation of preventive and control measures for intra-provincial spread of dengue fever. All data pertaining cross-county imported dengue fever cases within Yunnan Province in 2023 were collected, and the temporal, regional and population distributions of the cases were descriptively analyzed. A total of 1 664 intra-provincial cross-county imported dengue fever cases were reported in 95 counties (cities, districts) cross 16 profectures (cities) in Yunnan Province in 2023, accounting for 12.34% of total cases in the province. Cross-county imported dengue fever cases were predominantly reported during the period between August and October (1 516 cases, 91.11% of total cases), and peaked in September (659 cases), with a single-day peak on October 8 (36 cases). During the period from September 4 to 10, five counties (cities) with local dengue fever epidemics, including Jinghong City of Xishuangbanna Dai Autonomous Prefecture, Gengma Dai and Wa Autonomous County of Lincang City, Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, Mengla Coun ty of Xishuangbanna Dai Autonomous Prefecture, and Zhenkang County of Lincang City, exported 165 cross-county imported dengue fever cases to the rest of the province. Among the 1 644 intra-provincial cross-county imported dengue fever cases, the male to female ratio was 1.40∶1.00, and 1 329 cases were at ages of 15 to 55 years (79.87%), with farmers as the predominant occupation (886 cases, 53.25%). The top 5 counties (cities/districts) reporting the highest number of intra-provincial cross-county imported dengue fever cases included Simao District (266 cases) and Lancang Lahu Autonomous County (118 cases) of Pu'er City, Mengla County (91 cases) and Menghai County (91 cases) of Xishuangbanna Dai Autonomous Prefecture, and Mangshi City (73 cases) of Dehong Dai and Jingpo Autonomous Prefecture, which accounting for 38.40% of total imported cases. These intra-provincial cross-county imported dengue fever cases originated from 7 counties (cities/districts) in 4 prefectures (cities), including 1 261 cases (76.70%) from Jinghong City of Xishuangbanna Dai Autonomous Prefecture, 224 cases (13.63%) from Ruili City of Dehong Dai and Jingpo Autonomous Prefecture, 103 cases (6.27%) from Gengma Dai and Wa Autonomous County of Lincang City, 31 cases (1.89%) from Mengla County of Xishuangbanna Dai Autonomous Prefecture, 30 cases (1.82%) from Zhenkang County of Lincang City, 10 cases (0.61%) from Cangyuan Wa Autonomous County of Lincang City, and 5 cases (0.30%) from Mohan-Boten Economic Cooperation Zone of Kunming City. In addition, local dengue fever epidemics following intra-provincial cross-county importation of dengue fevers cases in Simao District, Jinggu Dai and Yi Autonomous County, Mangshi City, Longchuan County, and Cangyuan Wa Autonomous County. Farmers and students are high-risk populations for intra-provincial cross-county imported dengue fever cases in Yunnan Province, and health education pertaining personal protection against dengue fever should be strengthened among these high-risk populations by governments at all levels. There is a high risk of local out-break of dengue fever following continuous introduction of intra-provincial cross-county imported cases. Standardized management of intra-provincial cross-county imported dengue fever cases should be reinforced to reduce the risk of local epidemics. ［摘要］ 目的 分析2023年云南省登革热省内跨县 (市、区) 输入病例的流行病学特征, 为制定登革热省内扩散防控措 施和策略提供参考。方法 收集2023年云南省登革热省内跨县 (市、区) 输入病例资料, 对其报告时间、地区以及人群分 布特征进行流行病学描述分析。结果 2023年云南省16个州 (市) 95个县 (市、区) 累计报告省内跨县 (市、区) 输入病例 1 664例, 占当年该省登革热病例总数的12.34% (1 664/13 483) 。病例报告时间集中于8—10月, 该时段共报告1 516例 (占91.11%); 9月为报告病例数高峰 (659例), 10月8日为单日报告病例数高峰 (36例); 9月4—10日西双版纳傣族自治 州景洪市、勐腊县, 临沧市耿马傣族佤族自治县、镇康县以及德宏傣族景颇族自治州瑞丽市5个本地疫情县 (市、区) 向全 省输出跨县 (市、区) 病例165例。1 664例省内跨县 (市、区) 输入病例男女性别比为1.40∶1.00, 以15 ~ < 55岁病例数最 多 (1 329例, 占79.87%), 职业以农民为主 (886例, 占53.25%) 。省内跨县 (市、区) 输入病例报告数居前5位的分别是普 洱市思茅区 (266例) 、澜沧拉祜族自治县 (118例) 、西双版纳傣族自治州勐腊县 (91例) 、勐海县 (91例) 和德宏傣族景颇族 自治州芒市 (73例), 上述5个县 (市、区) 报告输入病例数占输入病例总数的38.40% (639/1 664) 。输入病例来源于4个州 (市) 7 个县 (市、区), 其中景洪市1 261 例 (占76.70%) 、瑞丽市224 例 (占13.63%) 、耿马傣族佤族自治县103 例 (占 6.27%) 、西双版纳傣族自治州勐腊县31例 (占1.89%) 、镇康县30例 (占1.82%) 、沧源佤族自治县10例 (占0.61%) 、昆明市 磨憨-磨丁经济合作区5例 (占0.30%) 。思茅区、景谷傣族彝族自治县、芒市、陇川县和沧源佤族自治县报告登革热省内 跨县 (市、区) 输入病例后均发生本地疫情。结论 农民和学生是云南省登革热省内扩散防控的重点群体, 应加强对这两 类人群的健康宣教。在跨县 (市、区) 输入性病例持续发生后, 本地登革热暴发风险较高, 故应加强输入病例的规范处置, 以降低本地疫情发生风险。.
To evaluate the effectiveness of the integrated soil-borne nematodiasis and clonorchiasis control programmesin Guizhou Province from 2019 to 2023, so as to provide insights into formulation of appropriate parasitic disease control strategies in the province. From 2019 to 2023, Shiqian County in Tongren City and Zhenfeng County in Qianxi'nan Buyi and Miao Autonomous Prefecture were selected as pilot counties in Guizhou Province for soil-borne nematodiasis prevention and control programmes, and Rongjiang County in Qiandongnan Miao and Dong Autonomous Prefecture was selected as a pilot county for clonorchiasis control programmes. Integrated control measures were implemented in these 3 pilot counties, including surveys on human parasitic infections, deworming, health education and improved water and sanitation. At least 1 000 individuals were sampled from each of three pilot counties using a stratified multi-stage random sampling method from 2019 to 2023 for detection of soil-borne nematodes and Clonorchis sinensis human infections, and the awareness of soil-borne nematodiasis and clonorchiasis control knowledge was investigated among residents in pilot counties using questionnaire surveys. In addition, the implementation of deworming and coverage of sanitary toilets and safe drinking water were collected in three pilot counties. The prevalence of soil-borne nematode human infections reduced from 7.78% (79/1 016), 2.80% (28/1 001) and 14.40% (144/1 000) in 2019 to 1.18% (12/1 014), 1.38% (14/1 001) and 2.73% (28/1 024) in 2023 in Shiqian County, Zhenfeng County, and Rongjiang County, respectively (χ2 = 51.51, 4.91 and 88.54, all P values < 0.05). No C. sinensis human infections were detected in Shiqian County or Zhenfeng County from 2019 to 2023, and the prevalence of C. sinensis human infections reduced from 1.80% (18/1 000) in 2019 to 0.29% (3/1 024) in 2023 in Rongjiang County (χ2 = 11.19, P < 0.05). Free deworming was provided to 574 cases with soil-borne nematode infections and 47 cases with C. sinensis infections detected in three pilot counties from 2019 to 2023. The coverage of health education was 100.00% in both Zhenfeng County and Shiqian County during the period from 2019 to 2023, and the awareness of soil-borne nematodiasis control knowledge increased from 93.60% (234/250) in Zhenfeng County and 70.97% (577/813) in Shiqian County in 2019 to 99.20% (248/250) and 98.40% (492/500) in 2023, respectively. The coverage of health education increased from 60.07% (161/268) in 2019 to 100.00% (250/250) in 2023 in Rongjiang County, and the awareness of clonorchiasis control knowledge increased from 80.67% (121/150) in 2019 to 99.20% (248/250) in 2023. The coverage of sanitary toilets increased from 48.89% (61 078/124 935), 34.20% (40 381/118 085) and 70.55% (60 604/85 920) in 2019 to 65.87% (77 649/117 878), 56.00% (63 252/112 948) and 89.15% (72 737/81 590) in 2023 in Shiqian County, Zhenfeng County and Rongjiang County, respectively, and the coverage of safe drinking water was all 100.00% in both Shiqian County and Rongjiang County during the 5-year study period, and increased from 85.33% (100 765/118 085) in 2019 to 100.00% (112 948/112 948) in 2023 in Zhenfeng County. There were remarkable reductions in the prevalence of soil-borne nematodes human infections in Shiqian County, Zhenfeng County, and Rongjiang County and in the prevalence of C. sinensis human infections in Rongjiang County, Guizhou Province following the 5-year integrated control programmes from 2019 to 2023. The wide-spread application of the health education-led and human parasitic diseases examination and treatment-based integrated soil-borne nematodiasis and clonorchiasis control programmes seems justified. ［摘要］目的 评价2019—2023年贵州省土源性线虫病和华支睾吸虫病综合防治措施实施效果, 为该省制定适宜的寄生虫病防控策略提供科学依据。方法 2019—2023年, 选择贵州省铜仁市石阡县、黔西南布依族苗族自治州贞丰县为 土源性线虫病防治试点县, 选择黔东南苗族侗族自治州榕江县为华支睾吸虫病防治试点县, 开展人群寄生虫感染情况调查、药物驱虫、健康教育、改水改厕等综合防治措施。2019—2023年3个试点县均采用分层多阶段随机抽样方法选择调查对象, 每个试点县均抽取不少于1 000人开展人群土源性线虫和华支睾吸虫感染调查。通过问卷调查了解试点县人群寄生虫病防治知识知晓情况。此外, 收集3个试点县药物驱虫实施情况以及卫生厕所普及率、安全饮用水普及率等数据。结果石阡县、贞丰县和榕江县人群土源性线虫感染率分别从2019年的7.78% (79/1 016) 、2.80% (28/1 001) 和14.40% (144/1 000) 下降至2023年的1.18% (12/1 014) 、1.38% (14/1 001) 和2.73% (28/1 024), 差异均有统计学意义 (χ2 = 51.51、4.91、88.54, P 均< 0.05) 。2019—2023年石阡县和贞丰县均未检出华支睾吸虫感染者; 榕江县人群华支睾吸虫感染率从2019年的1.80% (18/1 000) 下降至2023年的0.29% (3/1 024), 差异有统计学意义 (χ2 = 11.19, P < 0.05) 。对2019—2023年3个试点县查出的574例土源性线虫感染者和47例华支睾吸虫感染者均进行了免费驱虫治疗。贞丰县和石阡县自2019年起健康教育覆盖率均为100.00%, 人群土源性线虫病防治知识知晓率分别由2019年的93.60% (234/250) 和 70.97% (577/813) 上升至2023年的99.20% (248/250) 和98.40% (492/500) 。榕江县健康教育覆盖率从2019年的 60.07% (161/268) 上升至2023年的100.00% (250/250), 人群华支睾吸虫病防治知识知晓率由2019年的80.67% (121/150) 上升至2023年的99.20% (248/250) 。2019—2023年石阡县、贞丰县和榕江县卫生厕所普及率均有所提高, 分别从2019年的48.89% (61 078/124 935) 、34.20% (40 381/118 085) 和70.55% (60 604/85 920) 上升至2023年的65.87% (77 649/117 878) 、56.00% (63 252/112 948) 和89.15% (72 737/81 590); 石阡县和榕江县安全饮用水普及率自2019年起均为100.00%, 贞丰县从2019年的85.33% (100 765/118 085) 上升至2023年的100.00% (112 948/112 948) 。结论 经过5年综合防治, 贵州省石阡县、贞丰县和榕江县人群土源性线虫感染率以及榕江县人群华支睾吸虫感染率均显著下降, 以健康宣教为先导、 人群查治病为主的综合防治模式值得推广应用。.
To investigate the regulatory role of the programmed cell death protein 1 (PD-1) and its ligand programmed cell death protein ligand 1 (PD-L1) signaling on the subtypes and functions of natural killer (NK) cells at the maternal-fetal interface during the second trimester in mice following Toxoplasma gondii infection during the first trimester. Twelve 6- to 8-week-old female mice of the C57BL/6J strain were divided into a control group and an infection group, of 6 mice in each group. On the 6.5th day of pregnancy (Gd6.5), each pregnant mouse in the infection group was intraperitoneally injected with 150 tachyzoites of the Toxoplasma gondii PRU strain, while mice in the control group were injected with an equal volume of physiological saline. On the 12.5th day of pregnancy (Gd12.5), uterus and placenta tissues were sampled from pregnant mice for pathological observations, and the mRNA expression levels of PD-1, PD-L1, and tumor necrosis factor-α (TNF-α) were quantified in uterus and placenta tissues. The PD-1 and DX5 expression was measured on NK cells at the maternal-fetal interface using flow cytometry. In addition, the in vitro JEG-3 trophoblast cells and NK-92MI cells co-culture system was established as the control group, and the addition of T. gondii tachyzoites in the co-culture system served as the infection group. The PD-1, PD-L1, and DX5 mRNA expression was quantified in cells using real-time fluorescence quantitative reverse transcription PCR (RT-qPCR) assay, and the TNF-α concentration was measured in the cell culture supernatant using enzyme-linked immunosorbent assay (ELISA). On Gd12.5, clear and intact cellular structures of placental decidual tissues were seen in pregnant mice in the control group, with no remarkable abnormal changes found in the uterine columnar epithelial cells, and inflammatory cell infiltration and blood stasis at varying degrees were found in uterine and placental tissues from pregnant mice in the infection group. The relative PD-1, PD-L1, and TNF-α mRNA expression was (1.004 ± 0.004), (1.001 ± 0.001), and (1.001 ± 0.001) in uterine tissues from pregnant mice in the control group and (2.480 ± 0.720), (3.355 ± 0.920), and (2.391 ± 0.073) in the infection group, respectively. The relative PD-1, PD-L1, and TNF-α mRNA expression was (1.007 ± 0.010), (1.006 ± 0.006), and (1.001 ± 0.001) in the uterine tissues in the control group and (6.948 ± 1.918), (3.225 ± 1.034), and (1.536 ± 0.150) in the infection group, respectively. The relative PD-1, PD-L1, and TNF-α mRNA expression was higher in both the uterine (t = 3.55, 4.43 and 33.02, all P values < 0.05) and placental tissues (t = 5.36, 3.72 and 6.18, all P values < 0.05) in the infection group than in the control group. Flow cytometry showed that the proportions of PD-1+ NK cells, PD-1+ DX5+ NK cells, and DX5+ NK cells were (12.200 ± 1.082)%, (9.373 ± 7.728)%, and (44.000 ± 4.095)% in uterine tissues from pregnant mice in the control group, and (21.733 ± 1.630)%, (18.767 ± 1.242)%, and (73.367 ± 0.611)% in the infection group, respectively. The proportions of PD-1+ NK cells, PD-1+ DX5+ NK cells, and DX5+ NK cells were (1.100 ± 0.510)%, (2.277 ± 1.337)%, and (96.167 ± 2.831)% in placental tissues from mice in the control group, and (26.867 ± 9.722)%, (23.433 ± 6.983)%, and (82.467 ± 2.248)% in the infection group, respectively. The proportions of PD-1+ NK cells (t = 8.45, P < 0.05) and DX5+ NK cells (t = 12.29, P < 0.05) were higher in uterine tissues from pregnant mice in the infection group than in the control group, and no significant difference was seen in the proportion of PD-1+ DX5+ NK cells (Z = -1.09, P > 0.05). The proportions of PD-1+ NK cells (t = 4.58, P < 0.05) and PD-1+ DX5+ NK cells (t = 5.15, P < 0.05) were higher in placental tissues from pregnant mice in the infection group than in the control group, while the proportion of DX5+ NK cells was lower in the infection group than in the control group (t = -6.56, P < 0.05). RT-qPCR assay revealed that the relative PD-1, PD-L1, and DX5 mRNA expression was (1.010 ± 0.005), (1.002 ± 0.003), and (1.001 ± 0.001) in the JEG-3 cells and NK92MI cells co-culture system and (3.638 ± 1.258), (0.397 ± 0.158), and (4.267 ± 1.750) in the control group, and ELISA measured that the TNF-α concentration was higher in the cell culture supernatant in the infection group [(22.056 ± 3.205) pg/mL] than in the control group [(12.441 ± 0.001) pg/mL] (t = 5.20, P < 0.05). The PD-1(t = 3.62, P < 0.05) and DX5 mRNA expression (t = 3.23, P < 0.05) was higher in the infection group than in the control group, and the PD-L1 mRNA expression was lower in the infection group than in the control group (t = -6.63, P < 0.05). Following T. gondii infection, both PD-L1 expression and PD-1 expression on DX5+ NK cells at the maternal-fetal interface are upregulated in mice during the second trimester; however, the proportion of DX5+ NK cells decreases. These findings suggest that PD-1/PD-L1 signaling may suppress NK cell functions by modulating DX5+ NK cell subsets. ［摘要］ 目的 探讨小鼠孕早期感染弓形虫后, 孕中期母胎界面程序性死亡蛋白1 (programmed cell death protein-1, PD-1) 及其程序性死亡蛋白配体1 (programmed cell death protein ligand 1, PD-L1) 通路对自然杀伤 (natural killer, NK) 细胞亚型 和功能的调节作用。方法 将12只雌性C57BL/6J小鼠 (6~8周龄) 随机分为对照组和感染组, 每组6只。妊娠第6.5天 (Gd6.5) 时, 每只感染组孕鼠均采用腹腔注射方式感染150只弓形虫PRU株速殖子, 对照组小鼠腹腔注射等量生理盐水。 妊娠第12.5天 (Gd12.5) 时取孕鼠子宫和胎盘组织观察其病理变化, 检测组织中PD-1、PD-L1 及肿瘤坏死因子 (tumor necrosis factor, TNF) -α mRNA 表达水平, 并采用流式细胞术检测母胎界面NK细胞上PD-1和DX5表达水平。此外, 设置 JEG-3滋养层细胞与NK92MI细胞体外共培养体系为对照组, 在此基础上加入弓形虫速殖子作为感染组, 通过实时荧光 定量PCR (real-time quantitative reverse transcription polymerase chain reaction, RT-qPCR) 法检测细胞PD-1、PD-L1 及DX5 mRNA 表达水平, 采用酶联免疫吸附试验 (enzyme-linked immunosorbent assay, ELISA) 检测细胞培养上清液中TNF-α浓 度。结果 Gd12.5时, 对照组孕鼠胎盘蜕膜组织细胞结构清晰完整, 子宫柱状上皮细胞无明显异常改变; 感染组孕鼠子 宫和胎盘组织均出现不同程度炎症细胞浸润和淤血。对照组孕鼠子宫组织中PD-1、PD-L1、TNF-α mRNA 相对表达水平 分别为1.004 ± 0.004、1.001 ± 0.001、1.001 ± 0.001, 感染组分别为2.480 ± 0.720、3.355 ± 0.920、2.391 ± 0.073; 对照组胎盘 组织中PD-1、PD-L1、TNF-αmRNA相对表达水平分别为1.007±0.010、1.006±0.006、1.001±0.001, 感染组分别为6.948±1.918、 3.225 ± 1.034、1.536 ± 0.150; 与对照组相比, 感染组子宫组织 (t = 3.55、4.43、33.02, P 均< 0.05) 和胎盘组织中PD-1、PD-L1、TNF-α mRNA 表达水平 (t = 5.36、3.72、6.18, P 均< 0.05) 均升高。流式细胞术检测结果显示, 对照组孕鼠子宫组织中 PD-1+ NK细胞、PD-1+ DX5+ NK细胞和DX5+ NK细胞比例分别为 (12.200 ± 1.082) %、 (9.373 ± 7.728) %、 (44.000 ± 4.095) %, 感染组分别为 (21.733 ± 1.630) %、 (18.767 ± 1.242) %、 (73.367 ± 0.611) %; 对照组胎盘组织中PD-1+ NK细胞、PD-1+ DX5+ NK 细胞和DX5+ NK 细胞比例分别为 (1.100 ± 0.510) %、 (2.277 ± 1.337) %、 (96.167 ± 2.831) %, 感染组分别为 (26.867 ± 9.722) %、 (23.433 ± 6.983) %、 (82.467 ± 2.248) %。与对照组相比, 感染组小鼠子宫组织中PD-1+ NK细胞 (t = 8.45, P < 0.05) 、DX5+ NK细胞比例 (t = 12.29, P < 0.05) 升高, PD-1+ DX5+ NK细胞比例无显著变化 (Z = -1.09, P > 0.05); 胎盘组织中 PD-1+ NK细胞 (t = 4.58, P < 0.05) 、PD-1+ DX5+ NK细胞比例升高 (t = 5.15, P < 0.05), 而DX5+ NK细胞比例下降 (t = -6.56, P < 0.05) 。JEG-3细胞与NK92MI细胞体外共培养体系检测结果显示, 对照组细胞上PD-1、PD-L1、DX5 mRNA相对表达水 平分别为1.010 ± 0.005、1.002 ± 0.003、1.001 ± 0.001, 感染组分别为3.638 ±1.258、0.397 ± 0.158、4.267 ± 1.750; 对照组和感 染组细胞培养上清液中TNF-α浓度分别为 (12.441 ± 0.001) 、 (22.056 ± 3.205) pg/mL; 与对照组相比, 感染组PD-1 (t = 3.62, P < 0.05) 、DX5 mRNA (t = 3.23, P < 0.05) 表达均上调, PD-L1 mRNA 表达下调 (t = -6.63, P < 0.05), 且细胞培养上清 液中TNF-α浓度升高 (t = 5.20, P < 0.05) 。结论 刚地弓形虫感染后, 小鼠孕中期母胎界面中PD-L1表达水平及DX5+NK 细胞PD-1表达水平均上调、DX5+ NK细胞比例下降, 提示PD-1/PD-L1通路可能通过调控DX5+ NK细胞亚群抑制NK细胞 功能。.
To evaluate the value of serum Mac-2 binding protein (M2BP) for assessment of the severity of schisto somiasis-induced liver fibrosis, so as to provide insights into non-invasive diagnosis and disease surveillance of liver fibrosis caused by schistosomiasis. A total of 234 individuals with a history of Schistosoma japonicum infection were sampled from Xinhua Village, Lushan City, Jiangxi Province from 2019 to 2020, and 234 serum samples were collected from all participants. All participants received B-ultrasound examinations of the liver. Serum samples were categorized into four groups (grades 0, Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis groups) according to B-ultrasound examination results, and then, each group was randomly divided into a receiver operating characteristic (ROC) curve group and an efficacy assessment group at a ratio of 7∶3. Serum M2BP concentration was measured in four groups using the enzyme-linked immunosorbent assay (ELISA), and differences in serum M2BP concentrations were compared with analysis of variance and Spearman correlation analysis. Serum M2BP concentration was subjected to ROC curve analysis among individuals with different grades of schistosomiasis-induced liver fibrosis in the ROC curve group to determine the optimal diagnostic threshold of M2BP concentration at different fibrosis grades, and the area under the ROC curve (AUC) was calculated to evaluate the diagnostic performance. The diagnostic accuracy was verified by comparing the accordance rate and Kappa consistency test in the efficacy assessment group. Among 234 serum samples, there were 79 samples with grade 0 schistosomiasis-induced liver fibrosis, 87 samples with Grade Ⅰ, 46 samples with Grade Ⅱ and 22 samples with Grade Ⅲ according to the B-ultrasound examinations. The mean serum M2BP concentrations were (0.40 ± 0.31) [95% confidence interval (CI): (0.33, 0.47)], (0.64 ± 0.48) [95% CI: (0.53, 0.74)], (1.76 ± 0.58) [95% CI: (1.59, 1.93)] μg/mL and (2.56 ± 0.93) [95% CI: (2.14, 2.97)] μg/mL in the four groups, respectively (F = 150.796, P < 0.001), and the severity of schistosomiasis-induced liver fibrosis significantly positively correlated with serum M2BP concentration (rs = 0.715, P < 0. 001). The sample sizes of grades 0, Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis sera were randomly allocated as follows: 55 versus 24, 61 versus 26, 32 versus 14, and 15 versus 7 in the ROC curve and efficacy assessment groups, respectively, and the serum M2BP concentrations were (0.39 ± 0.29) μg/mL and (0.42 ± 0.36) μg/mL (F = 0.196, P > 0.05), (0.59 ± 0.47) μg/mL and (0.75 ± 0.51) μg/mL (F = 1.967, P > 0.05), (1.73 ± 0.59) μg/mL and (1.85 ± 0.57) μg/mL (F = 0.417, P > 0.05), and (2.46 ± 0.64) μg/mL and (2.76 ± 1.41) μg/mL (F = 0.491, P > 0.05), respectively. ROC curve analysis showed that the optimal diagnostic thresholds of serum M2BP concentration were 0.347 86 μg/mL (AUC = 0.635, P < 0.05), 1.188 83 μg/mL (AUC = 0.938, P < 0.000 1) and 2.021 21 μg/mL (AUC = 0.821, P < 0.000 1) for grade Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis. In addition, the accordance rates between the optimal diagnostic threshold of serum M2BP and B-ultrasound examinations for predicting grade Ⅰ, Ⅱ and Ⅲ schistosomiasis-induceed liver fibrosis were 69.23%, 85.71% and 71.43% (χ2 = 1.340, P > 0.05), and the overall Kappa consistency test showed moderate consistency [Kappa = 0.608, 95% CI: (0.428, 0.788); Z = 6.609, P < 0.000 1]. Serum M2BP may serve as a potential biomarker for assessing moderate to advanced schistosomiasis-induced liver fibrosis; however, its diagnostic value for early-stage schistosomiasis-induced liver fibrosis remains limited. ［摘要］ 目的 评估血清Mac-2结合蛋白 (Mac-2 binding protein, M2BP) 对血吸虫病肝纤维化严重程度的诊断价值, 为血 吸虫病肝纤维化的无创诊断和病情监测提供参考。方法 2019—2020年在江西省庐山市新华村抽取234名既往血吸虫 感染者作为调查对象, 并采集其血清样本, 共234份。对调查对象进行肝脏B超检查, 根据检查结果将血清样本分为血吸 虫病肝纤维化0、Ⅰ、Ⅱ级和Ⅲ级4个级别, 各级别再按7∶3的比例将样本随机分为受试者工作特征 (receiver operating characteristic, ROC) 曲线组和效能评估组。采用酶联免疫吸附法检测不同血吸虫病肝纤维化分级血清样本中M2BP浓 度, 并进行方差分析和Spearman相关性分析。对不同血吸虫病肝纤维化分级血清样本ROC组M2BP浓度进行ROC曲线 分析, 确定M2BP诊断不同肝纤维化分级的最佳诊断阈值, 并计算曲线下面积 (area under the curve, AUC) 以评估诊断效 能。根据不同血吸虫病肝纤维化分级所对应的M2BP最佳诊断阈值, 分别预测各效能评估组血清样本的肝纤维化分级, 并与B超检查结果进行符合率比较和Kappa一致性检验。结果 根据B超检查结果, 234份血清样本中血吸虫病肝纤维 化0、Ⅰ、Ⅱ级和Ⅲ级样本分别为79、87、46、22份, 不同肝纤维化分级样本血清M2BP平均浓度分别为 (0.40 ± 0.31) ［95% 可信区间 (confidence interval, CI) ： (0.33, 0.47) ］、(0.64 ± 0.48) ［95% CI： (0.53, 0.74) ］、(1.76 ± 0.58) ［95% CI： (1.59, 1.93) ］μg/mL和 (2.56 ± 0.93) ［95% CI： (2.14, 2.97) ］μg/mL, 差异有统计学意义 (F = 150.796, P < 0.001), 且血吸虫病肝纤 维化严重程度与血清M2BP浓度呈正相关 (rs = 0.715, P < 0.001)。血吸虫病肝纤维化0、Ⅰ、Ⅱ、Ⅲ级ROC曲线组和效能评 估组样本分别为55份和24份、61份和26份、32份和14份、15份和7份, 不同肝纤维化分级ROC曲线组和效能评估组血 清M2BP 浓度分别为 (0.39 ± 0.29) μg/mL 和 (0.42 ± 0.36) μg/mL、(0.59 ± 0.47) μg/mL 和 (0.75 ± 0.51) μg/mL、(1.73 ± 0.59) μg/mL和 (1.85 ± 0.57) μg/mL、(2.46 ± 0.64) μg/mL和 (2.76 ± 1.41) μg/mL, 差异均无统计学意义 (F = 0.196、1.967、0.417、0.491, P 均> 0.05)。血清M2BP诊断血吸虫病肝纤维化的ROC曲线分析结果显示, 其对肝纤维化Ⅰ、Ⅱ级和Ⅲ级 的最佳诊断阈值分别为0.347 86 (AUC = 0.635, P < 0.05)、1.188 83 μg/mL (AUC = 0.938, P < 0.000 1) 和2.021 21 μg/mL (AUC = 0.821, P < 0.000 1)。诊断效能评估结果显示, M2BP最佳诊断阈值对血吸虫病肝纤维化Ⅰ级、Ⅱ级和Ⅲ级的预测 结果与B超检查结果的符合率分别为69.23%、85.71%和71.43%, 差异无统计学意义 (χ2 = 1.340, P > 0.05), 总体一致性检 验呈中等程度一致［Kappa = 0.608, 95% CI： (0.428, 0.788); Z = 6.609, P < 0.000 1］。结论 血清M2BP可作为辅助诊断血 吸虫病肝纤维化中、晚期的潜在生物标志物, 但其对早期血吸虫病肝纤维化的诊断价值有限。.
To investigate the epidemiological characteristics of severe fever with thrombocytopenia syndrome (SFTS) and to identify factors affecting deaths among SFTS patients in China from 2010 to 2023, so as to provide insights into scientific prevention and control of SFTS. Demographic and epidemiological characteristics of reported, definitively diagnosed SFTS cases in China from 2010 to 2023 were captured from National Notifiable Infectious Disease Reporting System of China Information System for Disease Control and Prevention, including current residence address, age, gender, occupation, time of incidence and date of death, and the temporal, spatial and population distributions of SFTS cases were analyzed. The county level incidence of reported SFTS cases in China from 2010 to 2023 was subjected to spatial autocorrelation analysis, and the global Moran's I index was calculated. The high-incidence clusters for SFTS were identified using space-time scan analysis based on a Poisson distribution model, and the relative risk (RR) and log-likelihood ratio (LLR) were estimated. In addition, factors affecting the death and their risk levels were identified among SFTS cases using chi-square test and logistic regression models, and the risk of death was evaluated with odds ratio (OR). A total of 27 457 SFTS cases were reported in China from 2010 to 2023, and the number of SFTS cases increased from 71 in 2010 to 5 062 in 2023, appearing a tendency towards a rise (b = 5.567, t = 51.35, P < 0.05). A total of 1 326 deaths occurred during the 14-year study period, with a case fatality rate of 4.82%, and the annual incidence and fatality of SFTS were 0.005/105 to 0.359/105, and 2.70% to 12.70%. SFTS cases were reported across 27 provinces in China, which were predominantly reported in 7 provinces of Shandong (7 890 cases, 28.74%), Henan (6 286 cases, 22.89%), Anhui (5 718 cases, 20.83%), Hubei (3 938 cases, 14.34%), Liaoning (1 418 cases, 5.16%), Zhejiang (990 cases, 3.61%), and Jiangsu (957 cases, 3.49%), accounting for 99.05% (27 197/27 457) of totally reported cases in China. The time of SFTS incidence appeared a seasonal distribution, and the incidence of SFTS peaked during the period from May to July, with a significant difference in the time of SFTS incidence among provinces (P < 0.01). Among all SFTS cases, there were 12 894 men (46.96%) and 14 563 women (53.04%), and there were 61.27% (16 823/27 457) of SFTS cases at ages of 61 years and older, with farmers as the predominant occupation (84.74%, 23 266/27 457). The annual Moran's I index for SFTS incidence ranged from 0.326 2 to 0.607 5 from 2010 to 2023, and there were significant differences in the Moran's I index for SFTS incidence each year from 2011 to 2023 (Z = 10.207 to 18.101, all P values < 0.001), presenting spatial clusters. Local spatial autocorrelation analysis identified "high-high", "low-high", "high-low", and "low-low" clusters of reported SFTS cases in China, with "high-high" clusters predominantly distributed in 5 provinces of Shandong, Anhui, Hubei, Henan, and Liaoning, covering 63 counties (cities, districts) in 2011 to 134 counties (cities, districts) in 2023. Monthly space-time scans identified three high-incidence clusters for SFTS in 4 provinces of Henan, Shandong, Jiangsu and Anhui. Univariate analysis revealed the risk of death of reported SFTS cases was associated with province (χ2 = 605.482, P < 0.01), gender (χ2 = 23.421, P < 0.01), age (χ2 = 254.181, P < 0.01), duration from disease onset to diagnosis (χ2 = 49.895, P < 0.01), and occupation (χ2 = 30.685, P < 0.01), and multivariate logistic regression analysis revealed a higher risk of death among SFTS cases reported in three provinces of Shandong [OR = 3.081, 95% confidence interval (CI): (2.605, 3.643)], Zhejiang [OR = 4.280, 95% CI: (3.367, 5.441)], and Jiangsu [OR = 2.733, 95% CI: (2.059, 3.628)]; among SFTS cases at ages of 70 years and older [> 70 to 80 years: OR = 4.511, 95% CI: (1.626, 12.511); > 80 years and older: OR = 3.632, 95% CI: (1.241, 10.631)]; among male SFTS cases males than among female cases [OR = 1.243, 95% CI: (1.114, 1.387)]; and among SFTS cases 31 days and longer duration from disease onset to diagnosis [OR = 1.660, 95% CI: (1.254, 2.197)]. The number of reported SFTS cases has remarkably risen in China in recent years, with expanded geographic distributions, seasonal distribution and spatial clusters. Targeted preventive and control measures for SFTS are urgently needed. ［摘要］目的分析2010—2023年我国发热伴血小板减少综合征 (severe fever with thrombocytopenia syndrome, SFTS) 流行特征及病例死亡影响因素, 为该病科学防控提供参考。方法在中国疾病预防控制信息系统传染病报告信息管理系统中获取于2010—2023年报告且经终审的SFTS病例数据, 主要包括病例现住址、年龄、性别、职业、发病和死亡时间等, 分析SFTS报告病例流行病学特征。以县级行政区为单位, 对2010—2023年我国SFTS报告病例发病情况进行空间自相关分析, 计算全局Moran’s I 指数; 基于泊松分布模型对SFTS病例数据进行时空扫描分析, 计算相对危险度 (relative risk, RR) 、对数似然比 (logarithmic likelihood ratio, LLR) 。采用χ2检验和logistic回归模型分析SFTS病例死亡相关因素, 采用比值比 (odds ratio, OR) 表征死亡风险。结果2010—2023年我国累计报告SFTS病例27 457例, 报告病例数由2010年的71例增加至2023年的5 062例, 呈上升趋势 (b = 5.567, t = 51.35, P < 0.05); 各年发病率在0.005/10万~ 0.359/10万。上述SFTS报告病例中, 死亡1 326例, 病死率为4.82%; 各年病死率在2.70% ~ 12.70%。全国共有27个省份报告SFTS病例, 病例主要分布于山东省 (7 890例, 占28.74%) 、河南省 (6 286例, 占22.89%) 、安徽省 (5 718 例, 占20.83%) 、湖北省 (3 938例, 占14.34%) 、辽宁省 (1 418 例, 占5.16%) 、浙江省 (990例, 占3.61%) 和江苏省 (957例, 占3.49%), 上述7省报告病例数占我国SFTS报告病例总数的99.05% (27 197/27 457) 。病例发病时间具有季节性特征, 以5 ~ 7月为发病高峰; 不同省份病例发病时间分布差异有统计学意义 (P < 0.01) 。27 457例SFTS报告病例中, 男性12 894例 (占46.96%), 女性14 563例 (占53.04%) 。报告病例年龄以61岁及以上人群为主, 占61.27% (16 823/27 457); 职业以农民为主, 占84.74% (23 266/27 457) 。全局空间自相关分析结果显示, 2010—2023年我国SFTS发病率Moran’s I 指数值在0.326 2 ~ 0.607 5, 其中2011—2023年Moran’s I 指数值均有统计学意义 (Z = 10.207 ~ 18.101, P 均< 0.001), 病例分布存在空间聚集性。局部空间自相关分析结果显示, 我国SFTS报告病例存在“高-高”、“低-高”、“高-低”和“低-低”聚集区, 其中“高-高”聚集区主要分布于山东省、安徽省、湖北省、河南省及辽宁省, 聚集区范围由2011年63个县 (市、区) 增至2023年134个县 (市、区) 。逐月时空扫描分析发现3处SFTS报告病例高聚集区, 主要包括河南、山东、江苏省和安徽省4省。单因素分析发现, 我国SFTS病例死亡相关因素包括省份 (χ2 = 605.482, P < 0.01) 、性别 (χ2 = 23.421, P < 0.01) 、年龄 (χ2 = 254.181, P < 0.01) 、发病至诊断时间间隔 (χ2 = 49.895, P < 0.01) 及职业 (χ2 = 30.685, P < 0.01) 。多因素logistic回归分析结果表明, 山东省［OR = 3.081, 95% CI： (2.605, 3.643) ］、浙江省［OR = 4.280, 95% CI： (3.367, 5.441) ］和江苏省［OR = 2.733, 95% CI： (2.059, 3.628) ］SFTS报告病例死亡风险较高, 70岁以上组［> 70 ~ 80岁：OR = 4.511, 95% CI： (1.626, 12.511); > 80岁：OR = 3.632, 95% CI： (1.241, 10.631) ］病例死亡风险较高, 男性病例死亡风险高于女性［OR = 1.243, 95% CI： (1.114, 1.387) ］, 发病至诊断时间间隔≥ 30 d的病例死亡风险较高［OR = 1.660, 95% CI： (1.254, 2.197) ］。结论近年来我国SFTS报告病例数显著增加, 发病范围逐渐扩大, 发病具有季节和空间聚集性分布特征, 亟需制定有针对性的防控措施。.
As a novel healthcare management tool, diagnosis-related group (DRG) plays a significant role in standardization of medical behaviors, reduction of medical costs, and improvements of the utilization efficiency of medical resources. This article analyzes internal and external factors pertaining to the application of DRG in schistosomiasis control through a strengths, weaknesses, opportunities and threats (SWOT) analysis and discusses potential impacts, challenges, and development opportunities. DRG has shown significant potential in optimizing resources, standardizing diagnosis and treatment, and reducing expenses; however, it also suffers from problems of a great difficulty in grouping and a low informatization level at grassroots institutions. Optimization of diagnosis-related grouping and increased investments into grassroots institutions are therefore required to meet the special needs of schistosomiasis control, so as to improve the effectiveness of schistosomiasis control. ［摘要］ 疾病诊断相关分组 (diagnosis related groups, DRG) 作为一种新型医疗管理工具, 在规范医疗行为、控制医疗成本 和提高医疗资源利用率等方面效能显著。本研究通过采用优势、劣势、机会、威胁 (strengths,weaknesses, opportunities, threats, SWOT) 分析, 综合评估了DRG应用于血吸虫病防治领域的内、外部因素, 探讨其潜在影响、面临的挑战及发展机 遇。SWOT分析结果表明, DRG在优化资源配置、规范诊疗、减少费用支出方面具有较大潜力, 但也面临分组难度大、基 层信息化程度低等障碍。因此, 需优化DRG分组并加大基层投入, 使其更好地适应血吸虫病防治的特殊需求, 提升血吸 虫病防治效果。.
To investigate the scientific outputs of metagenomic next-generation sequencing (mNGS) for diagnosis of infection of unknown origin in intensive care units (ICUs), and to decipher the latest advances, frontier trends and spatiotemporal evolution of research hotpots in mNGS for diagnosis of infection of unknown origin in ICUs. Publications pertaining to the application of mNGS in diagnosis of infection of unknown origin in ICUs were retrieved from Web of Science Core Collection (WOSCC) from January 1, 2015 to December 31, 2024. The software Scimago Graphica 1.0.30 was employed to generate the network maps of collaboration relationships between countries, international collaborative relationships, author collaborations, institutional collaborative relationships, and a heatmap of journals, and the software VOSviewer 1.6.18 was used to create a heatmap of keywords, and maps of keyword co-occurrence clustering and keyword clustering timelines. In addition, the keyword burst map was created using the software CiteSpace 6.3.R3. A total of 1 707 publications were included in the final analysis, and the number of publications appeared an overall tendency towards a rise from 2015 to 2024, with the largest number of publications seen in 2024 (545 publications). The largest number of publications was recorded in China (1 390 publications), followed by in USA (190 publications) and United Kingdom (31 publications), and China led the global research in this field, with 81% of global related researches linked with China. Frontiers in Cellular and Infection and Microbiology published the largest number of articles (212 publications, 12.42%), and Joseph Derisi was the most productive author (33 publications). Author collaborations occurred within groups; however, there was a lack of close inter-group collaborations, with University of California, San Francisco and Chan Zuckerberg Biohub-based group seen as the largest collaborative group. High-frequency co-occurrence keywords included mNGS, infection, diagnosis, case report, community-acquired pneumonia and bronchoalveolar lavage fluid, and the 100 most common high-frequency co-occurrence keywords were assigned into four clusters. Keyword clustering timeline analysis revealed that the research hotspots in this field shifted from virus sequencing and sequence alignment to severe pulmonary infections, and keyword burst analysis showed identification, mNGS and virus as top three keywords with the highest burst intensity. mNGS was mainly used for identification of viruses among patients with infections of unknown origins in ICUs from 2015 to 2024, and future research priority shifted to pathogen detection for severe pulmonary infections. ［摘要］ 目的 了解宏基因组二代测序 (metagenomic next-generation sequencing, mNGS) 技术用于重症监护室 (intensive care unit, ICU) 不明原因感染诊断的科研产出, 揭示mNGS用于ICU不明原因感染诊断的最新进展、前沿趋势及研究热点 的时空演变。方法 在Web of Science核心数据集 (Web of Science Core Collection, WOSCC) 数据库中, 检索2015年1月1 日至2024年12月31日发表的mNGS用于ICU不明原因感染诊断的相关文献。采用Scimago Graphica 1.0.30软件绘制作 者所属国家及国际合作关系网络图、作者合作网络及研究机构合作关系网络图和期刊热图, 采用VOSviewer 1.6.18软件 绘制关键词热图、关键词共现聚类图以及时间叠加关键词聚类图, 同时使用CiteSpace 6.3.R3软件绘制关键词突现图。结果 共有1 707篇文献纳入分析, 2015—2024年发文量总体呈上升趋势, 以2024年发文量最高 (545篇)。中国发文量最 高 (1 390篇), 其次是美国 (190篇) 和英国 (31篇); 中国在该研究领域具有领先优势, 全球81%的相关研究均与中国相 关。以《Frontiers in Cellular and Infection and Microbiology (细胞与感染微生物学前沿) 》发文量最大 (212篇, 12.42%), Joseph Derisi博士产出最高 (33篇)。作者间合作主要存在于各个不同群组内, 群组间合作欠密切; 以加利福尼亚大学旧金 山分校和陈-扎克伯格生物中心为核心组成的合作群组规模最大。高频共现关键词包括mNGS、感染、诊断、病例报道、社 区获得性肺炎和支气管肺泡灌洗液等, 前100个高频共现关键词分为4个聚类; 时间叠加关键词聚类图显示, 该领域研究 重点从病毒测序和序列比对向严重肺部感染转变; 关键词突现图显示, 识别、mNGS和病毒是突现强度最高的前3个关键 词。结论 2015—2024年mNGS主要用于ICU不明原因感染病例病毒识别, 未来其研究重点将转向严重肺部感染病原 检测。.
Based on review of global and Chinese schistosomiasis control progress and the evolution of control strategies, this article focuses on Chinese practical experiences of schistosomiasis control and systematically summarizes five key determinants for schistosomiasis transmission risk, including source of infections, intermediate host snails, high-risk populations, natural environments, and social factors. To address these risks and challenges associated with these determinants, the article reviews the advances in techniques for assessment of schistosomiasis transmission risk and their applications, including conventional risk assessment approaches, mathematical model-based tools for prediction of schistosomiasis transmission risk, and indicator-systembased techniques for assessment of schistosomiasis transmission risk. This review underscores the essential role of interdisciplinary integration and dynamic management in precision schistosomiasis control and recommends the intensification of verification of field adaptation and dynamic updates of indicator systems to promote the widespread application of assessment tools across diverse regions and contexts, so as to provide strategic guidance and methodological support to achieve the target for elimination of schistosomiasis across China in 2030. ［摘要］ 本文在回顾全球及中国血吸虫病防控进展与策略演变的基础上, 聚焦中国实践经验, 系统梳理当前在血吸虫病 传播风险管控中传染源、中间宿主、重点人群、自然环境及社会因素等5大关键影响因素, 并针对上述风险与挑战, 从传 统风险评估方法、基于数理模型的传播风险预测方法与基于指标体系的风险评估技术3个方面, 对血吸虫病传播风险评 估技术及应用进展进行综述。本综述强调多学科融合与动态管理在精准防控中的作用, 并建议加强现场适应性验证与 指标体系动态更新, 推动评估工具在多区域、多情境下的广泛应用, 从而为实现2030年全国消除血吸虫病目标提供决策 参考与方法支撑。.
To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ2 = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system. ［摘要］ 目的 探究极端温度和干旱胁迫对球形小泡螺生存能力的影响, 为无参考基因条件下小泡螺基因组学研究提 供理论依据。方法 自桑给巴尔奔巴岛Kiwani Shehia地区现场采集球形小泡螺样本, 经实验室饲养、繁殖获取子代螺。选取120只来自同一代的10 周龄小泡螺作为实验螺, 随机分配至高温干旱组 (HD组)、常温干旱组 (D组)、低温干旱组 (LD组) 和对照组 (C组), 每组30只螺。将HD组、D组、LD组小泡螺均置于底部为干燥土壤的烧杯中, 随后分别置于35、26 ℃和10 ℃的气候培养箱中; C组小泡螺置于装有26 ℃去氯自来水的500 mL培养皿中。饲养3 d后收集各组活螺, 解 剖分离软体组织, 提取样本总RNA并构建相应基因库, 随后在Illumina HiSeq 4000 测序平台进行高通量测序。采用 Trinity 软件进行从头转录组组装并选取最长的转录本作为单基因簇。使用DIAMOND软件对单基因簇进行基因功能注 释, 用于基因功能标注的数据库包括基因本体论 (Gene Ontology, GO)、京都基因与基因组百科全书 (Kyoto Encyclopedia of Genes and Genomes, KEGG)、NCBI非冗余蛋白序列数据库 (NCBI non-redundant protein sequences database, NR)、蛋白质 家族数据库 (Protein Family Database, Pfam) 和瑞士-Prot蛋白序列数据库 (UniProtKB/Swiss-Prot, Swiss-Prot)。分别使用 topGO 软件和clusterProfiler软件对差异表达基因 (differentially expressed genes, DEGs) 进行GO、KEGG分析。另外选择4 个相关基因进行实时荧光定量PCR (real-time quantitative PCR, qRT-PCR) 验证, 以确定转录组测序结果的可靠性。结果 饲养3 d后, HD组、LD组、D组和C组分别有7、20、28只和30只小泡螺存活, 各组存活率分别为23.33% (7/30)、66.67% (20/30)、93.33% (28/30) 和100.00% (30/30), 差异有统计学意义 (χ2 = 52.72, P < 0.001)。通过无参转录组测序装 配了176 942个单基因簇, 在上述5个数据库中的标注率分别为0.98%、13.49%、26.46%、12.48%和14.39%。基因表达水 平分析结果显示, 与C组相比, D组共有33个基因上调、72个基因下调, HD组共有483个基因上调、815个基因下调, LD 组共有245个基因上调、172个基因下调。去除各组重叠基因及不匹配基因后, 共确定了11个候选基因。GO和KEGG分 析结果显示, 上述11个DEGs中, 热休克蛋白 (heat shock protein, HSP) 相关DEGs有3个, 分别注释为HSP12.2、HSP70、HSP20 基因, 在各处理组中的表达均显著上调; 免疫和神经系统相关DEGs有3个, 在各处理组中均显著下调, 涉及神经 细胞黏附分子L1样蛋白通路、纤维蛋白原结合蛋白通路和白细胞弹性蛋白酶抑制剂样蛋白通路。qRT-PCR检测结果显 示, 4个与温度及干旱胁迫相关的基因表达水平在不同处理组中的表达趋势与转录组测序结果高度一致。结论 极端 温度和干旱联合胁迫下球形小泡螺存活率降低, 可能与其细胞内稳态调节系统失衡有关。.