Bovine cysticercosis and taeniasis are significant neglected parasitic zoonotic diseases in sub-Saharan Africa, including Ethiopia. A cross-sectional study was conducted from October 2023 to May 2024 to estimate the prevalence of bovine cysticercosis at the Nekemte municipal abattoir and assess the level of public risk perception and practices. Ante-mortem examination and routine meat inspection were conducted, along with a questionnaire survey administered through face-to-face interviews to gather public responses on risk perception and practices related to bovine cysticercosis. Among the 402 cattle examined, 21.6% (95% CI: 17.7-26.0) were found positive for bovine cysticercosis. Age was a significant factor (p < 0.05) for the prevalence of bovine cysticercosis. Adult (aOR: 4.6, 95% CI: 1.3-15.7) and older (aOR: 3.8, 95% CI: 1.1-13.3) had higher odds of testing positive for bovine cysticercosis compared to young cattle. In terms of cyst distribution, the diaphragm, the shoulder muscle, and the heart were the most frequently affected organs, with prevalence rates of 6.5%, 5.0%, and 3.2%, respectively. A total of 178 cysts were recovered, resulting in an overall average intensity of 1.9 cysts per organ. The majority of respondents exhibited poor practices, 74.2% (95% CI: 65.4-81.7) and negative risk perception, 69.2% (95% CI: 60.1-77.3). Respondents who attended tertiary education were 4.8 times more likely to adopt good practices (OR = 4.8, 95% CI: 1.6, 14.1) compared to those who attended primary school or less. Urban residents demonstrate better practices, with a likelihood 3.3 times greater (OR = 3.3, 95% CI: 1.2-8.7) than rural residents. Muslims were 2.7 times more likely (OR = 2.7, 95% CI: 1.1, 6.3) to exhibit better practices compared to Christians. Furthermore, individuals with tertiary education had a risk perception that is 4.5 times more likely (OR = 4.5, 95% CI: 1.6, 11.6) than those who completed only primary school or less. In conclusion, the study revealed a moderately high prevalence of bovine cysticercosis, while public practices remained poor, coupled with a low level of risk perception. This highlights the need for increased attention to mitigate potential economic losses and health crises for both animals and humans associated with bovine cysticercosis.
Objective: To construct a reverse genetic system for the genotype ON1 of human respiratory syncytial virus subtype A (HRSV-A) expressing fluorescent reporter genes. Methods: Recombinant plasmids encoding EGFP or mCherry were constructed based on the 2019 Beijing HRSV-A ON1 dominant strain (6914). Recombinant viruses, rescued by co-transfecting BSR/T7-9 cells with helper plasmids, were identified via indirect immunofluorescence, whole-genome sequencing, and Western blot. Biological properties were characterized through fluorescent quantitative RT-PCR (qRT-PCR), immunostaining plaque assay and fluorescent focus assays (FFA). Results: Two recombinant viruses expressing EGFP or mCherry (rRSVA6914-EGFP and rRSVA6914-mCherry) were successfully rescued. Western blot analysis confirmed that the expression levels of key structural proteins (G, F, and N) in the recombinant strains were consistent with the parental virus. Multistep growth curve analysis revealed that the replication kinetics of the two recombinant viruses in HEp-2 cells did not differ significantly from those of the parental strain. Two recombinant viruses exhibited substantial neutralizing activity against both palivizumab and nirsevimab used in clinical settings. Furthermore, the viral titer of rRSVA6914-mCherry in A549 cells [(1.19±0.05)×105 PFU/ml] was significantly higher than in HEp-2 cells [(7.60±0.79)×104 PFU/ml] (P<0.001). For rRSVA6914-EGFP, the viral titers determined by immunostaining plaque assay and FFA methods were (1.15±0.17)×105 PFU/ml and (1.36±0.19)×105 FFU/ml. For rRSVA6914-mCherry, the corresponding titers were (3.50±0.23)×104 PFU/ml and (3.37±0.07)×104 FFU/ml. There was no statistically significant difference between the immunostaining plaque assay and FFA methods (both P>0.05). Conclusion: The HRSV-A genotype ON1 reverse genetic system expressing fluorescent reporter genes has been successfully constructed and systematically verified, providing a scientific tool for investigating the pathogenic mechanism of genotype ON1 and for screening antiviral drugs. 目的： 构建携带荧光报告基因的人呼吸道合胞病毒A亚型（HRSV-A）ON1基因型反向遗传学系统。 方法： 利用2019年北京地区分离的HRSV-A优势流行株ON1基因型6914株，构建携带增强型绿色荧光蛋白（EGFP）或红色荧光蛋白（mCherry）报告基因的重组质粒。通过与辅助质粒共转染至BSR/T7-9细胞拯救重组病毒；利用间接免疫荧光、全基因组测序、Western blot等方法鉴定病毒；基于荧光定量RT-PCR、免疫染色空斑试验及荧光灶形成试验（FFA）评价其生物学特性。 结果： 成功拯救出携带EGFP或mCherry报告基因的两株重组病毒（rRSVA6914-EGFP与rRSVA6914-mCherry）。Western blot证实重组病毒关键结构蛋白（G、F、N）表达水平与亲本株一致。多步生长曲线分析显示，两株重组病毒在HEp-2细胞中的复制动力学与亲本株基本一致。重组病毒对临床使用的帕丽珠单抗和尼塞韦单抗均表现出显著的中和活性。rRSVA6914-mCherry在A549细胞［（1.19±0.05）×105 PFU/ml］中的病毒滴度高于HEp-2细胞［（7.60±0.79）×104 PFU/ml］（P<0.001）。rRSVA6914-EGFP采用免疫染色空斑试验法和FFA法测得的病毒滴度为（1.15±0.17）×105 PFU/ml和（1.36±0.19）×105 FFU/ml，rRSVA6914-mCherry为（3.50±0.23）×104 PFU/ml和（3.37±0.07）×104 FFU/ml，两种方法测得的病毒滴度差异均无统计学意义（均P>0.05）。 结论： 成功构建并系统验证了携带荧光报告基因的HRSV-A ON1基因型反向遗传系统，可为ON1基因型致病机制研究及抗病毒药物筛选提供科学工具。.
Objective: To investigate the effects of miR-1910-3p on proliferation, invasion, epithelial-mesenchymal transition (EMT) and in vivo tumor growth of lung adenocarcinoma (LUAD) cells. Methods: Bioinformatics analysis was performed using The Cancer Genome Atlas (TCGA) database and the Starbase database to compare the expression of miR-1910-3p between LUAD tissues and adjacent normal tissues. In vitro experiments were conducted using human LUAD cell lines (H1299, A549) and normal lung epithelial cells (BEAS-2B). The expression level of miR-1910-3p was verified by real-time quantitative polymerase chain reaction (RT-qPCR). H1299 and A549 cells were transfected with liposomes to establish miR-1910-3p overexpression (mimic group), knockdown (inhibitor group) and negative control (NC group) cell models. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay and plate colony formation assay. Cell invasion was evaluated by Transwell invasion assay. The protein expression changes of EMT markers (E-cadherin, N-cadherin, Vimentin, Snail) were quantitatively analyzed by Western blot. For in vivo experiments, a subcutaneous xenograft model was established in nude mice, and tumor growth was monitored on days 7, 14, 21 and 28. At the end of the experiment, the nude mice were euthanized and the tumors were harvested for analysis. Results: Bioinformatics analysis showed that the expression level of miR-1910-3p was significantly higher in LUAD tissues than in adjacent normal tissues (P＜0.01). RT-qPCR results confirmed that compared with BEAS-2B cells, the relative expression levels of miR-1910-3p were significantly upregulated in A549 and H1299 LUAD cell lines (both P＜0.05). CCK-8 assay results showed that compared with the mimic NC group, overexpression of miR-1910-3p significantly enhanced the proliferative activity of A549 cells [mimic NC group (111.00±7.69)% vs. miR-1910-3p mimic group (119.70±7.54)%, P＜0.01] and H1299 cells [mimic NC group (113.40±15.21)% vs. miR-1910-3p mimic group (118.3±18.82)%, P＜0.01]; conversely, compared with the inhibitor NC group, knockdown of miR-1910-3p significantly inhibited the proliferative activity of A549 cells [inhibitor NC group (115.90±11.39)% vs. miR-1910-3p inhibitor group (111.90±8.83)%, P＜0.05] and H1299 cells [inhibitor NC group (113.20±15.34)% vs. miR-1910-3p inhibitor group (109.60±12.53)%, P＜0.05]. Plate colony formation assay showed that compared with the mimic NC group, overexpression of miR-1910-3p significantly enhanced the colony formation ability of A549 cells [mimic NC group (18.96±1.92)% vs. miR-1910-3p mimic group (37.33±3.66)%, P＜0.01] and H1299 cells [mimic NC group (22.86±2.78)% vs. miR-1910-3p mimic (42.33±2.58)%, P＜0.01]; conversely, compared with the inhibitor NC group, knockdown of miR-1910-3p significantly inhibited the colony forming ability of A549 cells [inhibitor NC group (19.46±3.33)% vs. miR-1910-3p inhibitor group (10.79±2.86)%, P＜0.01] and H1299 cells [inhibitor NC group (22.63±1.27)% vs. miR-1910-3p inhibitor group (12.96±1.45)%, P＜0.01]. Western blot analysis showed that compared with the mimic NC group, overexpression of miR-1910-3p significantly upregulated the expression of N-cadherin, Vimentin and the transcription factor Snail, while downregulating the expression level of E-cadherin (all P＜0.05). Transwell invasion assay showed that compared with the mimic NC group, overexpression of miR-1910-3p significantly enhanced the invasion ability of A549 cells [mimic NC group (333.00±35.68) vs. miR-1910-3p mimic group (521.67±46.92), P＜0.01] and H1299 cells [mimic NC group (341.67 ±32.87) vs. miR-1910-3p mimic group (537.66±33.13), P＜0.01]; conversely, compared with the inhibitor NC group, knockdown of miR-1910-3p significantly inhibited the invasive ability of A549 cells [inhibitor NC group (363.67±49.24) vs. miR-1910-3p inhibitor group (211.33±27.79), P＜0.01] and H1299 cells [inhibitor NC group (351.67±24.11) vs. miR-1910-3p inhibitor group (154.33±9.29), P＜0.01]. Subcutaneous xenograft experiment in nude mice showed that compared with the mimic NC group, overexpression of miR-1910-3p significantly promoted tumor growth in vivo, manifested as an increased tumor weight and volume (both P＜0.01). In the mimic NC group, tumor weights on days 7, 14, 21 and 28 were (0.08±0.01) g, (0.18±0.03) g, (0.41±0.06) g and (0.73±0.06) g, respectively; in the the miR-1910-3p mimic group, tumor weights on days 7, 14, 21 and 28 were (0.07±0.01) g, (0.35±0.06) g, (0.72±0.08) g, and (0.96±0.09) g, respectively. In the mimic NC group, tumor volumes on days 7, 14, 21 and 28 were (132.00±1.00) mm3, (254.67±7.10) mm3, (530.67± 42.71) mm3 and (853.33±74.10) mm3; in the miR-1910-3p mimic group, tumor volumes on days 7, 14, 21 and 28 were (132.00±2.00) mm3, (425.33±29.94) mm3, (829.00±62.00) mm3, and (1 123.33±95.38) mm3, respectively. Conclusion: miR-1910-3p acts an oncogene in LUAD, promoting tumor cell proliferation, invasion and in vivo tumorigenesis by activating EMT process. 目的： 探讨miR-1910-3p对肺腺癌细胞增殖、侵袭、上皮-间质转化（EMT）和体内肿瘤生长的影响。 方法： 通过癌症基因组图谱数据库和Starbase数据库进行生物信息学分析，分析miR-1910-3p在肺腺癌组织与癌旁正常组织中的表达差异。体外实验采用人肺腺癌细胞系（H1299、A549）及正常肺上皮细胞（BEAS-2B），应用实时荧光定量聚合酶链反应（RT-qPCR）验证miR-1910-3p表达水平。H1299和A549细胞分别通过脂质体转染法构建miR-1910-3p过表达（mimic组）、沉默（inhibitor组）及阴性对照（NC组）细胞模型。采用细胞计数试剂盒8（CCK-8）、平板克隆形成实验评估细胞增殖能力，Transwell侵袭实验检测细胞侵袭能力，Western blot技术定量分析EMT标志物（E-cadherin、N-cadherin、Vimentin、Snail）的蛋白表达变化。体内实验建立裸鼠皮下移植瘤模型，分别于第7天、第14天、第21天及第28天监测肿瘤生长情况，实验终点处死裸鼠并取瘤分析。 结果： 生物信息学分析显示，与癌旁正常组织比较，miR-1910-3p在肺腺癌组织中表达水平升高（P<0.01）。RT-qPCR验证结果显示，与BEAS-2B细胞比较，miR-1910-3p在肺腺癌细胞系A549细胞和H1299细胞中的相对表达水平均上调（均P<0.05）。CCK-8检测结果显示，与mimic NC组比较，过表达miR-1910-3p能够促进A549细胞［mimic NC组（111.00±7.69）%，miR-1910-3p mimic组（119.70±7.54）%，P<0.01］和H1299细胞［mimic NC组（113.40±15.21）%，miR-1910-3p mimic组（118.3±18.82）%，P<0.01］的增殖能力；反之，与inhibitor NC组比较，敲低miR-1910-3p能够抑制A549细胞［inhibitor NC组（115.90±11.39）%，miR-1910-3p inhibitor组（111.90±8.83）%，P<0.05］和H1299细胞［inhibitor NC组（113.20±15.34）%，miR-1910-3p inhibitor组（109.60±12.53）%，P<0.05］的增殖能力。平板克隆实验表明，与mimic NC组比较，过表达miR-1910-3p能够促进A549细胞［mimic NC组（18.96±1.92）%，miR-1910-3p mimic组（37.33±3.66）%，P<0.01］和H1299细胞［mimic NC组（22.86±2.78）%，miR-1910-3p mimic组（42.33±2.58）%，P<0.01］的克隆形成能力；反之，与inhibitor NC组比较，敲低miR-1910-3p能够抑制A549细胞［inhibitor NC组（19.46±3.33）%，miR-1910-3p inhibitor组（10.79±2.86）%，P<0.01］和H1299细胞［inhibitor NC组（22.63±1.27）%，miR-1910-3p inhibitor组（12.96±1.45）%，P<0.05］的克隆形成能力。Western blot分析结果显示，与mimic NC组比较，过表达miR-1910-3p能够同时上调N-cadherin、Vimentin及Snail转录因子的表达，下调E-cadherin的表达水平（均P<0.05）。Transwewll检测结果显示，与mimic NC组比较，过表达miR-1910-3p能够促进A549细胞［mimic NC组（333.00±35.68）个，miR-1910-3p mimic组（521.67±46.92）个，P<0.01］和H1299细胞［mimic NC组（341.67±32.87）个，miR-1910-3p mimic组（537.66±33.13）个，P<0.01］的侵袭能力；反之，与inhibitor NC组比较，敲低miR-1910-3p能够抑制A549细胞［inhibitor NC组（363.67±49.24）个，miR-1910-3p inhibitor组（211.33±27.79）个，P<0.01］和H1299细胞［inhibitor NC组（351.67±24.11）个，miR-1910-3p inhibitor组（154.33±9.29）个，P<0.01］的侵袭能力。裸鼠皮下成瘤实验显示，与mimic NC组比较，过表达miR-1910-3p能够促进裸鼠体内肿瘤生长，肿瘤重量和体积增加（均P<0.01）。mimic NC组裸鼠第7天、第14天、第21天及第28天肿瘤重量分别为（0.08±0.01）g、（0.18±0.03）g、（0.41±0.06）g和（0.73±0.06）g；miR-1910-3p mimic组分别为（0.07±0.01）g、（0.35±0.06）g、（0.72±0.08）g和（0.96±0.09）g。mimic NC组裸鼠第7天、第14天、第21天及第28天肿瘤体积分别为（132.00±1.00）mm3、（254.67±7.10）mm3、（530.67±42.71）mm3和（853.33±74.10）mm3；miR-1910-3p mimic组分别为（132.00±2.00）mm3、（425.33±29.94）mm3、（829.00±62.00）mm3和（1 123.33±95.38）mm3。 结论： miR-1910-3p在肺腺癌中发挥促癌基因作用，其通过激活EMT进程促进肿瘤细胞增殖、侵袭及体内成瘤。.
To investigate the expression pattern of methyltransferase-like protein 7B (METTL7B) in papillary thyroid carcinoma (PTC) and elucidate its role and molecular mechanisms in tumor metabolic reprogramming and malignant progression. Public datasets from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) were analyzed to assess METTL7B expression in PTC and its association with clinicopathological features. Immunohistochemistry on tissue microarrays was performed to validate METTL7B protein expression in clinical samples. Stable METTL7B-knockdown PTC cell lines were constructed, and knockdown efficiency was confirmed by quantitative real-time PCR (qRT-PCR) and Western blotting. Cell proliferation and clonogenic potential were evaluated using MTT and colony formation assays. Transcriptome sequencing was conducted in METTL7B-overexpressing cells to identify differentially expressed genes and enriched pathways, followed by qRT-PCR and Western blotting validation of METTL7B-mediated regulation of hypoxia-inducible factor 1α (HIF-1α). Cycloheximide chase and ubiquitination assays were used to assess the effect of METTL7B on HIF-1α protein stability, and the involvement of ubiquitin-specific protease 28 (USP28) was further examined. Lactate production and glucose uptake assays were performed to assess the impact of METTL7B and HIF-1α on PTC glycolytic activity. METTL7B was significantly upregulated in PTC tissues (P<0.05) and positively correlated with lymph node metastasis (P<0.05). METTL7B knockdown markedly suppressed PTC cell proliferation and colony formation (all P<0.05). In a xenograft tumor model, METTL7B knockdown significantly slowed the growth of PTC tumors, with tumor volumes significantly lower than those in the control group (P<0.05). Bioinformatics analysis suggested a strong association between METTL7B and the HIF-1α signaling pathway. Further experiments showed that METTL7B knockdown reduced HIF-1α protein levels without affecting its mRNA expression. Mechanistically, METTL7B inhibited the ubiquitination and degradation of HIF-1α, thereby maintaining its protein stability, and this process was mediated by the deubiquitinase USP28. Functional assays demonstrated that METTL7B enhanced glycolytic activity and malignant phenotypes of PTC cells by upregulating glycolysis-related genes through the USP28/HIF-1α axis. METTL7B promotes glycolysis and malignant progression in PTC by stabilizing HIF-1α through USP28-mediated deubiquitination. 目的: 探讨甲基转移酶样蛋白7B（METTL7B）在甲状腺乳头状癌（PTC）中的表达及其在肿瘤代谢重编程及恶性进展中的作用和分子机制。方法: 基于基因表达综合（GEO）和癌症基因组图谱（TCGA）数据库分析METTL7B在PTC组织中的表达水平及其与临床特征的关系。采用免疫组织化学染色验证METTL7B在临床组织芯片样本中的表达差异;构建METTL7B稳定敲减细胞株，并通过定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测敲减效率。进一步采用MTT法检测细胞增殖能力，克隆形成实验评估克隆形成能力。通过转录组测序分析METTL7B过表达后的差异基因并进行通路富集分析，结合免疫印迹法和qRT-PCR实验验证其对低氧诱导因子1α（HIF-1α）蛋白的调控作用。采用放线菌酮处理和泛素化检测评估METTL7B对HIF-1α蛋白稳定性的影响，并进一步探讨泛素特异性蛋白酶28（USP28）在其中的作用。乳酸生成及葡萄糖吸收实验分析METTL7B及HIF-1α对PTC糖酵解的调控。结果: METTL7B在PTC组织中显著高表达（P<0.05），并与淋巴结转移呈正相关（P<0.05）。敲减METTL7B显著抑制PTC细胞增殖及克隆形成能力（均P<0.05）。体内研究显示，敲减METTL7B可显著减缓PTC 裸鼠异种移植瘤的生长速度，并且肿瘤体积显著低于对照组（P<0.05）。生物信息学分析提示METTL7B与HIF-1α信号通路密切相关。进一步研究显示，敲减METTL7B降低HIF-1α蛋白而不影响其mRNA水平。机制研究表明，METTL7B可下调HIF-1α的泛素化降解并维持其蛋白稳定性，而去泛素化酶USP28可介导该过程。功能实验表明，METTL7B通过调控USP28/HIF-1α轴上调糖酵解关键基因表达，增强细胞糖酵解活性并促进其恶性生物学行为。结论: METTL7B通过USP28介导的去泛素化作用稳定HIF-1α，从而促进PTC的糖酵解和恶性进展。.
Objective: To investigate the expression of miR-527 in bladder urothelial cell carcinoma tissues (UCC) and cell lines, and to explore the mechanism by which miR-527 inhibits bladder cancer cell migration and promotes apoptosis through targeting Bcl-2. Methods: The expression of miR-527 in bladder UCC was analyzed using The Cancer Genome Atlas database. The expression of miR-527 in bladder UCC cell lines was verified by real-time quantitative reverse transcription polymerase chain reaction. The relationship between miR-527 and its target gene Bcl-2 was validated using a dual-luciferase reporter assay. miR-527 inhibitor and miR-527 mimic were constructed for cell transfection. Cell migration and apoptosis were assessed using wound healing assay and flow cytometry. Protein expression was detected by Western blot. In vivo tumorigenesis and the expressions of Bcl-2 and Ki-67 were evaluated using a nude mouse cell-derived xenograft model and immunohistochemistry. Results: Analysis of The Cancer Genome Atlas database revealed that miR-527 expression was significantly higher in bladder cancer tissues than in normal bladder tissues, whereas Bcl-2 protein expression was significantly lower. Dual-luciferase reporter assay confirmed that miR-527 directly targeted Bcl-2. Inhibition of miR-527 significantly upregulated Bcl-2 mRNA and protein levels (both P＜0.01), enhanced cell migration, and reduced apoptosis [migration rate: (100.00±0.00)% in the miR-527 inhibitor group vs. (47.74 ± 2.18)% in the T24 control group; apoptosis rate: (2.68±1.28)% vs. (11.82±5.19)%; both P＜0.05]. Overexpression of miR-527 produced opposite effects [migration rate: (45.22±3.66)% in the miR-527 overexpression group vs. (100.00±0.00)% in the UMUC-3 control group; apoptosis rate: (11.74±0.64)% vs. (3.81±0.75)%; both P＜0.05]. In animal experiments, overexpression of miR-527 significantly inhibited tumor growth and reduced the expressions of Bcl-2 and Ki-67 in tumor tissues, whereas inhibition of miR-527 promoted tumor growth, increased tumor cellular atypia, and enhanced the expressions of Bcl-2 and Ki-67. Conclusions: miR-527, as a tumor-associated gene, is highly expressed in bladder cancer tissues. Overexpression of miR-527 inhibits the growth and migration of bladder cancer cells and induces apoptosis by negatively regulating the Bcl-2. 目的： 探讨miR-527在膀胱尿路上皮癌组织和细胞系中的表达及其通过靶向Bcl-2抑制膀胱癌细胞迁移并促进癌细胞凋亡的机制。 方法： 利用癌症基因组图谱数据库分析miR-527在膀胱尿路上皮癌组织中的表达，通过实时荧光定量逆转录聚合酶链反应验证膀胱尿路上皮癌细胞系中miR-527的表达，双荧光素酶报告基因验证miR-527与靶基因Bcl-2的关系，构建miR-527 inhibitor和miR-527 mimic进行细胞转染，细胞划痕实验及流式细胞术检测细胞迁移和凋亡情况，Western blot检测相关蛋白表达，裸鼠细胞源性异种移植和免疫组化验证体内成瘤和Bcl-2、Ki-67表达情况。 结果： 通过癌症基因组图谱数据库分析，膀胱癌组织中miR-527表达显著高于正常膀胱组织，Bcl-2蛋白表达显著降低。双荧光素酶报告基因实验证实miR-527可直接靶向Bcl-2。抑制miR-527表达后，Bcl-2 mRNA及蛋白表达水平显著升高（均P＜0.01），细胞迁移能力增强，凋亡细胞减少［T24细胞miR-527抑制组、T24细胞对照组细胞迁移率：（100.00±0.00）%、（47.74±2.18）%，细胞凋亡率：（2.68±1.28）%、（11.82±5.19）%，均P＜0.05］，但过表达miR-527则相反［UMUC-3细胞miR-527过表达组和UMUC-3 细胞对照组细胞迁移率：（45.22±3.66）%、（100.00±0.00）%，细胞凋亡率：（11.74±0.64）%、（3.81±0.75）%，均P＜0.05］。在动物实验中，过表达miR-527显著抑制肿瘤生长，肿瘤组织中Bcl-2及Ki-67表达减弱；抑制miR-527则促进肿瘤生长，肿瘤细胞异型性增加，Bcl-2及Ki-67表达增强。 结论： miR-527作为一种肿瘤相关基因，在膀胱癌组织中高表达，同时过表达miR-527可通过靶向负调控Bcl-2基因抑制膀胱癌肿瘤细胞生长和迁移，并诱导肿瘤细胞凋亡。.
This study is aimed at investigating the therapeutic effects and the relevant mechanisms of Lactiflorin in ulcerative colitis (UC) via a combination of various methodologies. The PI3K/AKT pathway identified through network pharmacology, and key pathways and therapeutic effects were validated by animal experiments, molecular docking, and molecular dynamics simulations. The experimental verification was performed using an acute colitis model induced by 2.5% dextran sulfate sodium. This study further explored whether the key bioactive components could improve intestinal barrier integrity and alleviate ulcerative colitis by inhibiting the PI3K/AKT signaling pathway. We performed hematoxylin and eosin staining, cytometric bead array, western blotting, and immunofluorescence. Network analysis identified 718 predicted drug targets, among which 264 were related to UC therapeutic targets. Animal experiments further confirmed the significant role of the key pathways and the effects of the pharmacological intervention. Molecular docking and dynamics simulations demonstrated Lactiflorin with a strong binding affinity to STAT3 (- 11.24 kcal/mol), AKT1 (- 15.61 kcal/mol), and PIK3R1 (- 11.95 kcal/mol). In vivo experiments, Lactiflorin improved the Disease Activity Index score and histopathological score in UC-modelled mice and suppressed the expression of pro-inflammatory cytokines including IL‑23, IL‑12p70, IL‑17A, and IL‑1α. Western blot and immunofluorescence results revealed that Lactiflorin inhibited the expression of AKT, STAT3, and PI3K proteins (P < 0.01). These findings suggest that Lactiflorin exerts potential therapeutic effects against UC through PI3K/AKT pathway.
Scientific influence, or the capacity of ideas and concepts to shape future research, is crucial to developing and disseminating knowledge and sustained innovation. Here, using nearly 240 million scientific works published between 1990 and 2023 from OpenAlex to construct international networks of influence, discursive influence, which represents what global scientific communities consider important and worthy of investigation, is shown to be disproportionately and increasingly concentrated in a small group of resource-wealthy countries, including the United States, Canada, Western Europe and East Asia, in comparison to attributional influence. This concentration raises issues of equity and innovation in global scientific discourse, perhaps narrowing research perspectives, exacerbating biases and creating echo chambers that are associated with stifling innovation and marginalizing contributions from countries that are peripheral to global scientific discourse. The findings underscore the need to promote diverse and inclusive global research enterprises.
Metabolic dysfunction associated steatotic liver disease (MASLD) and its advanced form, MASH, are closely linked to cardiac dysfunction, particularly heart failure with preserved ejection fraction (HFpEF). However, the mechanisms underlying MASLD-associated HFpEF and its reversibility remain poorly understood, largely due to the lack of robust preclinical models. Here, we established a translational model of MASLD-associated cardiac dysfunction that recapitulates the key features of human HFpEF. We applied functional and transcriptomic analyses of the left ventricle (LV) to define the pathways associated with cardiac dysfunction and its reversibility. Alms1-/- (Foz/Foz) mice and wild-type littermates were fed normal chow (NC) or Western diet (WD) for up to 36 weeks (wk). Reversibility was modeled by switching WD-fed Foz/Foz mice at 12wk back to NC for 12wk. Cardiac assessment included echocardiography, invasive hemodynamics with dobutamine stimulation, histopathology, electron microscopy and isolated cardiomyocyte contractility. LV transcriptomes were profiled by bulk RNA sequencing and analyzed by differential expression and pathway enrichment. Foz/Foz mice on WD for 24wk developed metabolic syndrome and MASH with advanced liver fibrosis. Cardiac phenotyping showed LV hypertrophy, impaired cardiomyocyte contractility, reduced β-adrenergic reserve, elevated plasma BNP, and increased mortality while the ejection fraction was preserved (>50%), consistent with HFpEF. The progression of cardiac dysfunction was closely associated with liver fibrosis that developed during MASH. Switching WD-fed Foz/Foz mice at 12wk to normal chow diet reversed hepatic fibrosis, restored LV function, and reduced mortality, demonstrating plasticity of the liver-heart axis. LV transcriptomic analysis revealed that cardiac impairment in these mice was associated with mitochondrial dysfunction, altered substrate utilization, extracellular matrix remodeling, and metabolic stress; pathways that are similarly dysregulated in human HFpEF. Cardiac electron microscopy revealed swollen mitochondria with disrupted cristae, which improved following dietary intervention. Mitochondrial dysfunction and fibroinflammatory remodeling are prominent features of MASLD-associated cardiac dysfunction. Reversal of hepatic and cardiac phenotypes with dietary intervention, together with elucidation of underlying pathways, establish the Foz/Foz model as a useful translational platform for studying liver-heart axis in MASLD.
Emerging infectious diseases (EIDs) cause significant health and economic burdens in the USA and globally. Existing methods and analyses fall short of what is required to prioritise diseases for health technology research and development (R&D), including for medical countermeasure (MCM) development within rapid response frameworks by the Center for Biomedical Advanced Research and Development Authority, part of the Administration for Strategic Preparedness and Response within the U.S. Department of Health and Human Services. We developed a method for quantifying and ranking health and economic disease burdens ('full burdens') and applied it to 15 high-priority EIDs for 223 countries and territories, including the USA and US territories, historically from 2000 to 2022 and prospectively from 2025 to 2034. Health burdens consisted of disability-adjusted life-year losses, converted into monetary values using the value of a statistical life-year. Economic burdens consisted of direct and indirect costs during the acute stage of illness for hospitalised cases. We computed unweighted and weighted burden measures, the latter controlling for global disparities in ability-to-pay to avoid EID burdens. We projected future disease burdens using Monte Carlo simulation. Pandemics caused the largest historical and projected unweighted and weighted full burdens in the USA and globally. Among non-pandemics, across unweighted and weighted burdens, dengue and cholera imposed the largest historical and projected full burdens globally; West Nile Virus imposed the largest historical and projected full burdens in the USA, and dengue imposed the largest historical full burdens in the US territories. Weighted full burdens exceeded five times the unweighted ones. Regionally, the Americas and Africa faced the largest per capita weighted burdens while the Western Pacific region faced the smallest. R&D priority-setting, including MCM development, depends on multiple criteria, including disease burdens. Our full burden quantification methods and results, along with other such criteria, can inform optimal priority-setting.
Gastric cancer (GC) is one of the leading causes of global cancer mortality, underscoring an urgent need for novel therapeutic targets. Dysregulation of ubiquitination and deubiquitination plays a critical role in tumorigenesis. The deubiquitinating enzyme FAM105A, belonging to the OTULIN subfamily, has not yet been characterized in cancer. Preliminary bioinformatics analysis indicated that FAM105A is upregulated in gastric adenocarcinoma and positively correlates with the expression of CTGF, a downstream target of the Hippo/YAP signaling pathway, which is frequently dysregulated in GC. This study aimed to elucidate the role and mechanism of FAM105A in GC progression. Bioinformatics databases were used to analyze the expression of FAM105A and its clinical significance, which was further validated in clinical samples via immunohistochemistry (IHC) and Western blot (WB). In vitro and in vivo functional assays were conducted using stable FAM105A-knockdown and FAM105A-overexpressing GC cell lines. The interaction between FAM105A and YAP was investigated using co-immunoprecipitation (Co-IP), immunofluorescence, ubiquitination assays, and proximity ligation assay (PLA). The functional dependency on the Hippo pathway was assessed using the inhibitor XMU-MP-1. Domain mapping was performed by constructing truncated mutants of both FAM105A and YAP. FAM105A was significantly overexpressed in GC tissues, and its high expression correlated with advanced TNM stage and poor prognosis. Functionally, FAM105A promoted GC cell proliferation, migration, and inhibited apoptosis in vitro. In vivo evaluation using xenograft models demonstrated that FAM105A-overexpressing xenografts exhibited significantly larger tumor volumes compared to controls, while FAM105A-knockdown tumors showed reduced growth. FAM105A knockdown significantly decreased final tumor weight, whereas overexpression increased tumor mass. Immunohistochemical analysis revealed that knockdown tumors exhibited consistently reduced Ki-67 immunopositivity, while overexpression samples showed enhanced proliferation signatures. Mechanistically, Co-IP experiments further confirmed that FAM105A binds to YAP via its OTU domain, whereas full-length YAP, but not its truncation variants, efficiently interacts with FAM105A. FAM105A deubiquitinates and stabilizes YAP protein, promotes its nuclear translocation, and thereby activates the Hippo/YAP signaling pathway. The oncogenic effects of FAM105A overexpression were effectively reversed by the Hippo pathway inhibitor XMU-MP-1. This study provides comprehensive in vitro and in vivo evidence demonstrating that the deubiquitinating enzyme FAM105A functions as an oncogene in GC by stabilizing YAP and activating the Hippo/YAP pathway, thereby promoting tumor growth. FAM105A represents a promising prognostic biomarker and a potential therapeutic target for gastric cancer.
Human toxocariasis is a parasitic infection caused by Toxocara species and is important in the differential diagnosis of patients presenting with unexplained eosinophilia. A case of toxocariasis with an eosinophilic pleural effusion is presented. A 67-year-old Japanese man presented with progressive dyspnea. Laboratory investigations showed marked peripheral eosinophilia and a markedly elevated serum immunoglobulin E level. Chest imaging demonstrated bilateral ground-glass opacities and a left-sided pleural effusion. Analysis of the pleural fluid showed eosinophil predominance without evidence of malignancy or bacterial infection. Extensive diagnostic evaluations, including autoimmune serology, bone marrow examination, and genetic analyses for myeloid neoplasms with eosinophilia, yielded negative results. Serological test using an enzyme-linked immunosorbent assay demonstrated elevated anti-Toxocara antibody level, which were subsequently confirmed by western blot analysis, leading to a diagnosis of human toxocariasis presenting with visceral larva migrans. The patient had no clear history of animal contact or ingestion of raw meat or liver. Treatment with oral albendazole for four weeks resulted in marked clinical improvement, with resolution of the pleural effusion and pulmonary infiltrates, normalization of peripheral eosinophil counts, and a significant decrease in anti-Toxocara antibody level. This case emphasizes the importance of considering toxocariasis in the differential diagnosis of an eosinophilic pleural effusion and pulmonary infiltrates, even in the absence of identifiable exposure risks. Early serological testing can facilitate prompt diagnosis and appropriate anthelmintic therapy, potentially preventing unnecessary invasive procedures and delayed treatment.
To develop and validate a deep learning (DL) model for automatic quantification of knee effusion-synovitis volume (ESV) on MRI, assess correlations of ESV with semiquantitative effusion-synovitis (sqES) scores, and compare associations of ESV and sqES with MRI features and symptoms of knee osteoarthritis. A DL model was developed to quantify ESV on baseline right-knee MRI from the Osteoarthritis Initiative (n=4,698), which was trained and tested using manual segmentations from 101 randomly selected knees. Dice coefficients were used to quantify segmentation performance. Spearman correlations were computed between ESV and sqES scores from Whole-Organ Magnetic Resonance Imaging Score (WORMSES) and MRI Osteoarthritis Knee Score (MOAKSES). Paired differences in standardized β coefficients (Δβstd) from linear models were used to compare associations of ESV with tissue-specific WORMS scores and the total score of the Western Ontario McMaster Universities Arthritis Index (WOMAC). The DL model achieved a mean Dice coefficient of 0.79 (95% Confidence Interval [CI] 0.71-0.86) on the test set. ESV showed moderate correlations with WORMSES (ρ=0.50, 95% CI 0.47-0.53) and MOAKSES (ρ=0.65, 95% CI 0.63-0.67) and demonstrated larger effect sizes for associations with WORMS features (minimum Δβstd=0.03 [95% CI 0.00-0.07]) and knee OA symptoms (minimum Δβstd=0.03 [95% CI 0.00-0.07]) than sqES scores. A DL model for automated quantification of knee ESV on MRI was developed and validated. While our results demonstrate the potential of ESV as a scalable imaging biomarker in osteoarthritis research, validation in independent cohorts is necessary to confirm its utility.
Apoptosis plays a paramount role in endometriosis pathogenesis. This process may be disrupted in endometrial stromal cells (ESCs) of women with endometriosis, causing them to continue developing in ectopic locations. This study investigates the role of apoptosis in endometriosis by comparing the protein expression of Fas (CD95) and Fas ligand (FasL) in ESCs of women with endometriosis to that of healthy controls. Additionally, it examines the gene expression levels of Fas and FasL in peritoneal fluid mononuclear cells (PFMCs) and peripheral blood mononuclear cells (PBMCs) from both groups. Lastly, it assesses the levels of soluble FasL (sFasL) released by ESCs and PFMCs. ESCs were isolated from ectopic (n = 11) and eutopic samples (n = 17) of endometriosis patients and control (n = 10), alongside peritoneal fluid and blood samples from 10 patients and 10 controls. Using Western blot, Fas and FasL protein expression were assessed in ectopic endometrial stromal cells (EESCs), eutopic endometrial stromal cells (EuESCs), and control endometrial stromal cells (CESCs). Additionally, quantitative real-time PCR was used to evaluate Fas and FasL gene expression in PFMCs and PBMCs. Lastly, an enzyme-linked immunosorbent assay (ELISA) was conducted to assess the concentration of sFasL molecules in the supernatants of EESCs, EuESCs, CESCs, and PFMCs. Importantly, Fas protein levels in EESCs and EuESCs were lower than in CESCs (p < 0.01). Conversely, FasL protein levels were elevated in EESCs compared to both EuESCs and CESCs (p < 0.01 and p < 0.05, respectively). Additionally, patients with endometriosis exhibited higher Fas gene expression in PFMCs (p < 0.05) and lower expression in PBMCs (p < 0.01) compared to controls, along with reduced FasL expression in PFMCs (p < 0.01). Moreover, the concentration of sFasL molecules released from EESCs was significantly higher compared to EuESCs (p < 0.01) and CESCs (p < 0.05). All in all, our findings shed light on the understanding of the involvement of the Fas/FasL pathway in endometriosis pathogenesis.
House dust mites (HDMs), particularly Dermatophagoides farinae, are commonly found in household dust and play a key role in allergic diseases such as asthma and allergic rhinitis. Beyond clinical management, allergen removal strategies are crucial for improving quality of life. Hence, this study investigated the effects of ozone exposure on D. farinae, focusing on changes in protein expression, surface bacterial composition, mortality, and mobility. Mites were exposed to ozone concentrations of 0.05, 0.5, and 1 ppm for 24, 48, and 72 h in a controlled chamber, with non-exposed mites serving as controls. Western blotting using anti-Der f 1 and anti-Blo t 5 antibodies assessed changes in allergen profiles, while 16 S rRNA sequencing characterised changes in surface bacterial communities. Mortality was evaluated using 100 mites per group under varying exposure durations. To assess behavioural responses, a three-chamber mobility assay was conducted, where mites were placed in a central compartment flanked by no-ozone and low-ozone chambers, and their distribution was recorded after 72 h. Ozone exposure resulted in a concentration- and time-dependent reduction of Der f 1 protein intensity, suggesting allergen degradation. Surface bacterial profiling revealed distinct compositional shifts following ozone exposure. Mortality increased proportionally with ozone concentration and duration. In the mobility assay, mites predominantly remained in the no-ozone chamber, indicating avoidance of ozone. Collectively, these findings demonstrate that ozone exposure affects D. farinae at molecular, microbial, and behavioural levels, highlighting ozone's potential role in modulating mite allergenicity and ecology.
This scoping review aims to map and describe mental health indicators used in peer-reviewed studies in the WHO European Region to inform the development of a regional mental health measurement framework. reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Review (PRISMA-ScR), this scoping review analysed 75 studies identified through database searches on mental health monitoring and indicators in the WHO European Region. The scope was restricted to English-language, peer-reviewed literature published from 2019 onwards. Indicators were extracted and standardised to reflect their original reported meaning, treated as distinct and pragmatically classified into nine predefined domains. Across the 75 studies, 450 distinct indicators were identified. Most indicators related to mental health status and mental health risk factors and/or determinants. Geographic coverage varied, with northern and western European countries most frequently represented among the included literature. Diverse measurement tools were employed. The predominance of indicators related to mental health status and determinants reflects the emphasis on burden and underlying factors, while system-level or structural indicators remain under-represented. Differences in geographic coverage likely stem from disparities in research capacity, publication practices and data availability. This scoping review provides a descriptive overview of mental health indicators applied in recent peer-reviewed research in the Region. Greater alignment between research indicators and policy frameworks may strengthen the comparability and policy utility of mental health data across the Region.
Glauconite-bearing sandstone reservoirs represent promising targets for potential subsurface CO2 storage via mineral trapping. Nevertheless, previous studies commonly indicate that substantial CO2 sequestration through glauconite carbonation occurs on a geological timescale. In this study, we present results of a controlled experimental assessment of CO2 mineral trapping in Albian glauconite-bearing sandstones from the Mangyshlak Basin (southwestern Kazakhstan). The studied unit, composed of very fine to fine-grained sandstones, is commonly 30-40 m thick and extends hundreds of kilometers laterally. A 30-day CO2 injection batch experiment was performed at 100°C and 150 bar using a brine/rock ratio of 20. The aqueous phase exhibited a sustained increase in Fe, Na, K, Mg, and Si ions, followed by a modest late-stage decline in Mg, Fe, and Si ions, implying mineral dissolution followed by ion consumption through formation of secondary minerals. These results are supported by petrographic comparison of pre- and post-experiment mineral assemblages, which confirm the dissolution of glauconite clasts, feldspars, albite, and K-feldspar overgrowths, with minor precipitation of ankerite. Although the experiments clearly demonstrate significant dissolution of glauconite, the precise mechanisms governing the partitioning and incorporation of Fe and Mg into newly formed carbonate and/or secondary phyllosilicate phases require further mineralogical and geochemical investigations. The current results demonstrate that nascent laboratory scale glauconite carbonation can be feasible under high temperature-pressure conditions and significant concentration of Fe2+.
Genotype × environment interaction (GEI) poses a major challenge to the identification of stable, high-yielding capsicum genotypes under heterogeneous agro-climatic conditions. The present study evaluated 96 diverse capsicum (Capsicum annuum L.) genotypes across four staggered transplanting environments during 2022 and 2023 in the North-western Himalayas to dissect GEI for fruit yield and its component traits. Combined analysis of variance revealed highly significant (p ≤ 0.01) effects of genotype, environment, and GEI for all traits, indicating substantial genetic variability and differential environmental responses. AMMI analysis effectively partitioned interaction effects, with the first two interaction principal component axes (79-95%) explaining a large proportion of GEI for most traits. GGE biplot analysis provided clear visualization of genotype performance and stability. Genotypes G5 and G6 consistently exhibited superior marketable fruit yield with wide adaptability, while several genotypes showed specific adaptation to individual environments. The GGE biplot interpretation revealed that environment E3 was the most representative for selecting stable genotypes, while E2 demonstrated high discriminative ability among genotypes. The integrated AMMI-GGE approach facilitated the identification of promising genotypes under the evaluated environments at the study location, demonstrating its utility for stability assessment in capsicum.
Cancer stem cells (CSCs) serve as critical drivers of cancer relapse, metastasis and drug resistance, and are closely associated with poor prognosis. Increasing evidence has highlighted the regulatory effects of CSCs on immune cells such as macrophages in the tumor microenvironment (TME). Therefore, it is imperative to thoroughly study the specific mechanisms by which cancer stemness traits modulate macrophages. Transcriptomic and clinical data of STAD were retrieved from The Cancer Genome Atlas (TCGA). A prognostic Lasso-Cox model was constructed based on CSC-related genes using the R package 'glmnet'. Pathway enrichment analysis was performed using the 'clusterProfiler' package. Composition estimation of infiltrating immune cells in STAD tissues was conducted by CIBERSORTx. Western blotting and flow cytometry were used to detect the expression of CSC-related and macrophage polarization-related markers. Through analysis of the TCGA-STAD dataset, 35 CSC-related genes were upregulated in tumor tissues and associated with shorter survival, whereas 4 CSC-related genes were downregulated and correlated with longer survival. These 39 genes were used to construct the prognostic model, from which an optimal 17-gene CSC-related signature was derived and used to stratify patients into high-risk and low-risk groups. The high-risk group was related to poorer prognosis than the low-risk group in both training and testing cohorts. Within this model, CXCR4 exhibited the highest regression coefficient and was significantly associated with poor prognosis in STAD patients. Furthermore, CXCR4 inhibition significantly attenuated CSC-like properties in STAD cells and reduced expression of the CSC markers CD44 and CD24. Immune infiltration analysis revealed that the proportion of M2 macrophages was significantly increased, while M1 macrophages were decreased in the high-risk group. Moreover, CXCR4 expression was positively correlated with hypoxia-inducible factors and glycolysis regulators, and CXCR4 facilitated M2 macrophage polarization and migration by regulating lactate secretion. We established and validated a CSC-related prognostic model that was closely associated with macrophage polarization and clinical outcomes in STAD. CXCR4 was identified as the key gene in this model, which unregulated lactate secretion to drive M2 macrophage polarization and maintain CSC properties in STAD, and may serve as a putative regulatory factor in STAD progression. The CXCR4 inhibitor AMD3100 exerted significant anti-tumor effects in vitro, suggesting that CXCR4 may serve as a promising prognostic biomarker and a candidate target for STAD.
Type 2 diabetes mellitus and depression are mutually reinforcing, yet combined pharmacotherapy is limited by side effects and poor tolerability. Transcutaneous auricular vagus nerve stimulation (taVNS) is a non-invasive neuromodulatory approach with emerging metabolic and antidepressant potential, although its central mechanisms remain unclear. Here, we investigated whether hypothalamic serotonergic signaling is involved in the dual effects of taVNS in a mouse model of type 2 diabetes with depression (T2DD). T2DD was induced in db/db mice by chronic unpredictable mild stress, followed by taVNS intervention (2/15 Hz, 1 mA, 30 min/day for 3 weeks). Behavioral performance, body weight, and fasting blood glucose were assessed, while brain-wide neuronal activity was mapped using fluorescence micro-optical sectioning tomography (fMOST). Hypothalamic 5-HT release was monitored by fiber photometry, and 5-HT content and 5-HT₁A receptor expression were quantified by ELISA and Western blotting. TaVNS significantly reduced body weight and fasting blood glucose, alleviated depression-like behaviors, and improved locomotor activity. Brain-wide mapping revealed state-dependent modulation of neuronal activity, characterized by widespread activation under physiological conditions and suppression of stress-induced hyperactivation in T2DD. Notably, taVNS induced robust hypothalamic 5-HT release, increased long-term 5-HT levels, and upregulated 5-HT₁A receptor expression. Correlation analyses further linked enhanced hypothalamic serotonergic signaling to improved glycemic control and behavioral outcomes. These findings suggest that hypothalamic 5-HT signaling may contribute to the dual metabolic and antidepressant effects of taVNS and highlight its potential as an integrative neuromodulatory intervention for metabolic-psychiatric comorbidity.
Thalictrum glandulosissimum (TG) is traditionally used for treating ulcerative colitis (UC). Given the continuously increasing global incidence of UC and the limitations of current therapeutic options, there is an urgent need for safer treatments. However, most existing studies have focused on single component. Studies on the multi-component profile of TG's active alkaloids, particularly systematic pharmacokinetic investigations under pathological conditions, remain insufficient. Without a systematic understanding of which components are absorbed into the bloodstream and how their disposition is altered under UC pathology, the clinical translation of TG is significantly hindered. This research first established a UPLC-Q-TOF-MS/MS method to characterize TG's chemical composition, identifying 58 constituents including 16 prototypes. Integrating network pharmacology and in vitro assays, we predicted berberine, coptisine, cryptopine, and berlambine as core components, with STAT3, EGFR, and SRC as key targets. Molecular docking, qRT-PCR, and Western blot were used for validation. Furthermore, a rapid UPLC-MS/MS method was developed and validated to quantify five major bioactive alkaloids in rat plasma. Pharmacokinetic comparisons between normal and UC model rats revealed that UC significantly shortened Tmax (0.5-1.0 h) and increased the Cmax of berberine, coptisine, and columbamine by 1.19- to 1.67-fold (p < 0.05), suggesting enhanced intestinal absorption under inflammation. This study completes a "prediction-verification-application" workflow, laying a solid foundation for further pharmacokinetic-pharmacodynamic investigations into the therapeutic effects of TG in UC.