Better evaluation of the contribution of the main diseases, injuries, and risk factors for mortality and life expectancy is crucial for more efficient policy making at the national and subnational levels in Iran. The aim of this study is to assess the effect of emerging causes of mortality on health, specifically COVID-19, which can help policy makers implement preventive measures in similar situations. In this systematic analysis of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023, we present estimates of cause-specific mortality at the national and subnational levels in Iran from 1990 to 2023. New to this iteration of GBD, we present a decomposition analysis of the contribution of specific causes of death to net gain or loss in life expectancy across 31 provinces of Iran. We used an array of data sources including censuses, vital registration, and surveys for national and subnational estimates. The two leading causes of death in Iran were ischaemic heart disease and stroke in both 1990 and 2019. However, in 2020 and 2021, the COVID-19 pandemic displaced the leading causes of death, ranking first with age-standardised mortality rates of 286·2 deaths (95% uncertainty interval 267·9-310·5) per 100 000 in 2020 and 250·0 deaths (233·2-272·5) per 100 000 in 2021. COVID-19 ranked second and tenth in 2022 and 2023, respectively. Life expectancy at birth for both sexes combined declined from 78·0 years (77·7-78·1) in 2019 to 74·3 years (74·0-74·4) in 2020. It steadily recovered to 78·8 years (78·5-79·2) in 2023. COVID-19 was the main cause of loss in life expectancy, by 4·19 years, between 2019 and 2020. There was a net gain of 12·4 years in life expectancy in Iran from 1990 to 2023. The net gain at the national level can be mostly attributed to reduced mortality from ischaemic heart disease (2·61 years), stroke (1·63 years), neonatal disorders (1·26 years), transport injuries (0·88 years), and neoplasms (0·64 years). The decline in mortality rates of major causes continued to 2023 despite the pandemic. An exception was Alzheimer's disease, which showed a 4·0% increase in rate between 2019 and 2023 and led to a net loss of 0·04 years in life expectancy since 1990. Diabetes led to a net loss of 0·09 years since 1990. There were variations between provinces in terms of age-standardised rates and the net change in life expectancy before and after the COVID-19 pandemic. The COVID-19 pandemic disrupted the rising trend of life expectancy in Iran, varying across provinces. Findings show that the health-care infrastructure and policies in Iran were not efficient in controlling the pandemic in 2020 and 2021, mainly due to inadequate vaccination coverage and timeliness, specifically for vulnerable subgroups. Sanctions may have aggravated the effect of COVID-19 on loss in life expectancy of Iranians. Despite the pandemic, the declining trend in age-standardised rates for top causes of mortality has continued to 2023, leading to a full recovery of life expectancy and underscoring the ultimate resilience of Iran's health system. Gates Foundation.
To identify the continuous dose-response relationship between the duration of premature rupture of membranes (PROM) and the probability of neonatal and maternal infectious morbidity. This meta-analysis and systematic review synthesise data from 15 studies worldwide involving more than 70,000 mother-neonate pairs. A two-step random-effects model of PROM duration as a continuous dose, using restricted cubic splines, was used to estimate specific risk thresholds. The analysis established a progressive, non-linear escalation of risk. The onset of statistical risks at 16 hours is the early-onset pneumonia (Adjusted OR 1.86, 95% CI: 1.152.99). At the age of 18 hours, the incidence of culture-proven sepsis in neonates was 4.0%, and the odds ratio for maternal fever was significantly higher (AOR 36.6). The analysis of the ROC curves revealed a critical mathematical pivot point at 37 hours, after which complications escalate exponentially. Latency greater than 48 hours was the most significant independent predictor of culture-proven sepsis, with an increased risk of 8.2 (p < 0.001). Histologic chorioamnionitis was detected in 39% of mothers, and in many cases, they are clinically silent. Considerable heterogeneity (I2 > 60%) was mainly caused by gestational age disparities in cohorts of extremely preterm and term babies. PROM latency risk is not a threat but accelerates with time. Although 18 hours will be an acceptable early warning level, the range of 37 to 48 hours is a high-risk period that needs aggressive treatment. International guidelines need to be reviewed to reflect this non-linear trend, especially regarding pregnancy, where the risks of delivery are low compared to the rising risk of latency.
Quantitative detection of human immunodeficiency virus-type 1 (HIV-1) and hepatitis C virus (HCV) RNA plays a crucial role in the diagnosis, monitoring of the therapy and evaluation of the treatment response. The ELITe BeGenius® platform (ELITechGroup, Turin, Italy) is a fully automated sample-toresult molecular system integrating extraction, amplification and detection within a single workflow. The HIV-1 ELITe MGB® and HCV ELITe MGB® assays are real-time polymerase chain reaction tests designed for plasma viral-load quantification. This study aimed to verify their analytical performance under routine clinical laboratory conditions. Verification included assessments of accuracy, intra- and inter-assay precision, linearity and method correlation. A total of 70 plasma samples for HIV-1 RNA and 52 for HCV RNA were analyzed using previously tested and stored patient specimens, reference materials, and external quality controls. Results were compared with established reference assays used in accredited laboratories. Statistical analyses included positive, negative, and overall percent agreement (PPA, NPA, OPA), coefficients of variation (CV%), correlation and regression analyses and Bland-Altman bias estimation. For HIV-1 RNA, 19 of 20 positive and all 20 negative plasma samples were correctly identified by the ELITe MGB® assay, yielding a PPA of 95.0%, NPA of 100.0% and OPA of 97.5% (κ= 0.95). Intra-assay precision showed strong repeatability, with CVs of <1-3.9% for low-positive, 0.4-6.4% for medium-positive and <2% for high-positive specimens. Inter-assay reproducibility was consistent with CVs of 12.8% at low, 2.4% at medium, and 1.4% at high viral loads. Correlation analysis showed excellent concordance with the reference assay (p= 0.975, p< 0.001; R²= 0.95) and a mean bias of -0.40 log10 copies/mL in Bland-Altman analysis. Linearity was strong (R²= 0.97), confirming accurate quantification across the dynamic range with minor underestimation at higher dilutions. For HCV RNA, all 14 positive and 14 negative samples were correctly classified (PPA, NPA, and OPA= 100%; κ= 1.00). Intra-assay precision was excellent, with CVs around 2% for both low- and medium-positive samples, confirming consistent repeatability within a single run. Inter-assay reproducibility was equally robust, with CVs of 0.4-3.3% for low positives, 1.0-2.5% for medium and <1.1% for high-titer specimens. Correlation with the comparator method was strong (r= 0.956, p< 0.001; R²= 0.91) with a mean bias of -0.38 log10 IU/mL. Linearity analysis confirmed high proportionality between expected and measured concentrations (R²= 0.96). Deviations were negligible at low titers and slightly elevated at high loads but remained within acceptable limits. The HIV-1 and HCV ELITe MGB® assays on the BeGenius® platform demonstrated high accuracy, reproducibility and linearity, showing excellent correlation with reference methods. These results confirm that the ELITe BeGenius® system provides reliable and clinically valid viral-load measurements suitable for routine diagnostic use. Comprehensive laboratory verification of molecular assays under real-world conditions is crucial to ensure consistent performance, cross-platform comparability and reliable viral-load monitoring.
Introduction. Chronic rhinosinusitis is a difficult-to-treat, recurrent inflammatory condition of the nose and paranasal sinuses with global prevalence. Despite its impact on patient quality of life and its cost to the healthcare system, the pathogenesis of chronic rhinosinusitis (CRS) remains poorly understood. Additionally, while the presence of bacteria in CRS has been confirmed by numerous studies, their influence on disease symptoms is unclear. Disease-relevant models can help resolve these questions.Hypothesis. We hypothesized that bacterial inoculation could drive CRS-associated symptoms in a murine model.Aim. To characterize host-microbe interactions in a murine model of sinonasal bacterial infection.Methodology. Staphylococcus aureus and/or Pseudomonas aeruginosa were inoculated in the nasal cavity of Swiss Webster, C57Bl/6, Balb/c and B6.Cg-Prkdc scid/SzJ severe combined immunodeficient (SCID) mice. Systemic cytokine response was quantified with a multiplexed enzyme-linked immunosorbent assay, and local histological alterations were quantified using haematoxylin and eosin as well as Alcian Blue-Periodic Acid-Schiff-stained sinonasal sections.Results. Intranasal bacterial inoculation induced symptoms of CRS in murine sinonasal cavities. Dual species inoculation generated a unique response compared to single species. Repeated inoculations did not result in bacterial clearance from immunological priming. While Swiss Webster and C57Bl/6 mice demonstrated the greatest magnitude of responses, Balb/c mice demonstrated a protective response, generally downregulating cytokines and attempting to prevent further tissue damage. SCID mice demonstrated effective clearance of P. aeruginosa by innate immunity, but maintenance of S. aureus.Conclusion. Pathogenic bacteria are able to persist and drive the development of symptoms associated with clinical CRS in a murine model. Bacterial interactions and host factors influence CRS-associated inflammation. By investigating host responses from a number of mouse genetic backgrounds, the heterogeneity of disease presentation in CRS can be modelled, and strategies for infection management can be evaluated as potential therapeutic targets.
Human flora-associated (HFA) mice are often used to simulate the structure of human intestinal microbiota and to study the causal relationships between diseases and gut microbiota. However, several factors affect the colonization efficiency of human microbiota in germ-free (GF) mice, and the differential effects of gavage and lower gut transplantation on colonization are still unclear. In this study, we explored the reproducibility of the recipient-to-donor gut microbiota community structure and function under different transplantation routes and the differences in microbial colonization between recipients via gavage transplantation (GT_mice group) and lower gut transplantation (LGT_mice group). High-throughput sequencing of the metagenome was performed on the feces of each subject, and the composition of microbiome of each group was analyzed. As expected, the introduction of human fecal microbiota into GF mice via lower gut transplantation had a high transfer efficiency, which was evident from the similar species community structure to that of the donor (Adonis R2=0.713 960 for LGT_mice group‒donor group; Adonis R2=0.774 095 for GT_mice group‒donor group) and a higher bacterial colonization rate. The findings provide unique insights into improving the accuracy of constructing humanized microbiota transplantation models, aiding our understanding of the relationships between the human gut microbiota and disease. 人类菌群相关（HFA）小鼠常被用于模拟人类肠道微生物群结构，以探索疾病与肠道微生物群间的因果关系。然而，人类微生物群在无菌小鼠中的定植效率受多种因素影响，且灌胃和灌肠移植两种途径的定植效果差异尚待阐明。本研究探讨了不同移植途径对受体与供体间肠道微生物的群落结构和功能的重现性的影响，并比较了灌胃（上消化道移植，GT_mice组）和灌肠（下消化道移植，LGT_mice组）两种菌群移植方式在受体小鼠中微生物定植差异。我们通过对每个受体的粪便进行高通量宏基因组测序，并分析各组微生物组组成。结果显示，通过下消化道移植将人类粪便微生物群引入无菌小鼠具有较高的转移效率，这不仅体现在其物种群落结构与供体相似度较高（Adonis分析结果:LGT_mice组-供体组，Adonis R2=0.713 960;GT_mice组-供体组，Adonis R2=0.774 095），也表现为更高的细菌定植率。上述发现为提高人源化微生物群移植模型的构建准确性提供了新见解，有助于加深对人类肠道微生物群与疾病关联机制的理解。.
Respiratory failure is a serious complication of community-acquired pneumonia (CAP) that threatens children's health. In this study, the clinical characteristics of CAP with respiratory failure caused by different pathogens were analyzed. The distribution characteristics of the infectious pathogens were analyzed. Further statistical analysis of clinical data was conducted among the Mycoplasma pneumoniae (MP), respiratory syncytial virus (RSV), and bocavirus (BoV) groups. A total of 16 pathogens were detected, with the top three in terms of infection frequency being MP, RSV, and BoV. The duration of illness prior to hospitalization was longer in the multiple-pathogen group compared with the single-pathogen group. The duration of illness prior to hospitalization and the length of hospital stay were longer in the MP group. The distribution of abnormal imaging findings observed via radiographic examination among the MP, RSV, and BoV groups was related to the course of the illness. The MP group required the longest duration of oxygen therapy, with the majority of cases exceeding 7 days. In children, MP, RSV, and BoV are common pathogens associated with CAP accompanied by respiratory failure. In CAP with respiratory failure, differences in the infecting pathogens lead to variations in clinical characteristics.
Pneumonia remains the leading cause of mortality and morbidity among pediatric age groups worldwide. Several risk factors contribute to the progression of pneumonia into severe and complicated forms. Parapneumonic effusion is one of the most common complications to consider when severe pneumonia advances to a more critical stage. This study evaluates the prevalence, risk factors, and disease progression of severe pneumonia in children, providing valuable insights for resource-limited settings. A hospital-based prospective cohort study was conducted at a tertiary center from July 2022 to December 2023 among children aged 2 months to 14 years with severe pneumonia. Data were collected on socio-demographic characteristics, clinical status, disease progression, microbiological findings, and patient outcomes, and were analyzed using Stata Version 14. A total of 74.13% of severe pneumonia cases occurred in children under 5 years of age. Fever, cough, tachypnea, dyspnea, and chest indrawing were the most frequently observed clinical presentations. Overall, 31.46% of patients progressed to parapneumonic effusion. Absence of prior pneumonia (AOR = 0.25; 95% CI: 0.11-0.57), no prior hospitalization for the current diagnosis (AOR = 0.49; 95% CI: 0.28-0.86), no exposure to secondhand cigarette smoke (AOR 0.24; 95% CI: 0.07-0.84), and no contact with a coughing patient (AOR = 0.53; 95% CI: 0.29-0.96) were each associated with lower odds of progression to parapneumonic effusion. In contrast, malnutrition was associated with higher odds of progression (AOR = 2.00; 95% CI: 1.18-3.54). A higher prevalence of severe pneumonia was observed in children under 5 years of age. Disease progression to parapneumonic effusion occurred in 31.46% of cases, and the mortality rate was 4.54%.
The burden and clinical correlates of dementia are unknown in Uganda. We therefore determined the prevalence and factors associated with dementia among older persons in Northern Uganda. In a cross-sectional study, adults aged 60 years and above attending the medical outpatient clinic at a tertiary Hospital from March 2022 to April 2022 were enrolled. The prevalence of dementia was determined using the Intervention for Dementia in Elderly Africans (IDEA) and the Instrumental Activities of Daily Living (IADL) tool henceforth presented as IDEA-IADL. Factors of association were collected using a standardized questionnaire. A total of 271 eligible participants were enrolled into the study. Majority were age 60-69 years (153/271) and of male gender (165/271). The prevalence of dementia was 17.7% (48/271), severe dementia 7.4%, (20/271), moderate dementia 10.3%, (28/271). Factors independently associated with dementia were: participants aged between 80-89 years; (adjusted odds ratio (aOR): 9.7; 95% Confidence Interval (CI): 4.08-23.07, p< 0.001), family history of dementia (aOR: 3.4, 95CI: 1.62-7.04, p=0.001). history of depression (aOR: 2.7, 95CI: 1.40-5.02, p=0.003). Physical activity and cognitive exercise were associated protective factors with (aOR 0.1, 95CI: 0.03-0.24, p< 0.001) and (aOR 0.2, 95CI: 0.06-0.53, p<0.002) respectively. There burden of dementia among older persons in Northern Uganda is significant. Enhanced screening and early identification of dementia is recommended in this setting.
Autophagy plays a crucial role in maintaining cellular homeostasis and has been implicated in the pathogenesis of knee osteoarthritis (OA). However, data on radiographic stage-dependent transcriptional variation of autophagy-related genes in patients with knee OA, particularly using peripheral blood samples, remain limited. The aim of this study was to evaluate whether disease severity was associated with stage-dependent changes in the expression of selected autophagy-related genes within a patient cohort. A total of 200 patients diagnosed with knee OA were included in the study. Disease severity was classified according to the Kellgren-Lawrence radiographic grading system. Peripheral blood samples were collected, and the expression levels of selected autophagy-related genes were analyzed using quantitative real-time polymerase chain reaction [autophagy-related 5 (ATG5), ATG7, unc-51-like kinase 1 (ULK1), microtubule-associated protein 1 light chain 3 beta (LC3B), WD repeat domain phosphoinositide-interacting protein 1 (WIPI1), neighbor of BRCA1 gene 1 (NBR1), forkhead box O3 (FOXO3), transcription factor EB (TFEB)]. Relative gene expression was calculated using the ΔCt method, and comparisons were performed across radiographic stages. Associations between gene expression levels and systemic inflammatory markers were also assessed. Significant stage-dependent differences were observed in the expression of ULK1, TFEB, WIPI1, and NBR1 (p<0.05), with higher ΔCt values (reduced relative expression) in advanced radiographic stages compared with early-stage disease. In contrast, ATG5, ATG7, LC3B, and FOXO3 expression remained stable across radiographic stages. Furthermore, no significant associations were observed between expression of autophagy-related genes and systemic inflammatory status, as assessed by C-reactive protein levels. In patients with knee OA, regulatory and early autophagy-related genes exhibit radiographic stage-associated transcriptional alterations in peripheral blood, while expression of core autophagy machinery genes remain relatively stable. These findings suggest that disease severity is associated with selective transcriptional changes in autophagy-related pathways within the OA patient population and support further investigation of stage-dependent molecular patterns in knee OA.
During the COVID-19 pandemic, international border restrictions, along with traveler screening and quarantine, were implemented to limit virus spread. This study analyzes the epidemiological and genomic profiles of SARS-CoV-2 infections imported into Kinshasa (DRC) during the restrictions period in 2021. As part of the national response to the pandemic, self-reported demographic and clinical data were collected from travelers entering the DRC via N'djili-Kinshasa International Airport. SARS-CoV-2 infection was diagnosed using RT-PCR, and positive samples were subjected to whole genome sequencing (WGS) to determine variant types and viral lineages. The impact of the virus's genomic profile on the clinical presentation of travelers and on the COVID-19 epidemiology in the DRC was then assessed. Of 102,810 included travelers, 1037 (1.0%) tested positive for SARS-CoV-2 and reported significantly more nausea, diarrhea, and weight loss than uninfected travelers (p < 0.001). SARS-Cov-2-infected travelers were predominantly under 43 years old (p < 0.001) and primarily from France (24.8%) and Belgium (19.5%). Of the 105 WGS analyzed, 86 (81.9%) were variants of concern (VOCs), 14 (13.3%) were variants under monitoring (VUM), and the main genomic lineages identified were Delta-B.1.617.2 (24.8%), Alpha-B.1.1.7 (10.5%), Delta-AY.122 (7.6%), and B.1.620 (5.7%). The Delta-VOC was the most prevalent among positive travelers (61/86) and appeared to cause more symptomatic infections than non-Delta variants, although one-third of positive travelers reported no symptoms. SARS-CoV-2 importation into Kinshasa (DRC) mirrored global variant circulation patterns at the study's time. This genomic landscape was consistent with in-country clinical observations, emphasizing the importance of robust border surveillance and adaptive public health strategies during pandemics.
Streptococcus suis is a serious zoonotic pathogen responsible for rapid progression and deadly infections in both humans and pigs. With an increasing number of reported cases and considering the limitations of standard routine identification, a simple, rapid, and cost-effective approach is needed. In this study, a label-free colorimetric assay based on gold nanoparticles (AuNPs) was applied with a specific aptamer, R8-su12. This assay offered simplified detection through observable color change, enabling visual analysis by the naked eye or assessment via UV-Vis spectrophotometry. Under the optimal assay conditions, the detection procedure was carried out within 45 min. The reaction of the aptasensor and other bacterial species, including Staphylococcus aureus, S. pneumoniae, S. pyogenes, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium, and E. faecalis, was not present, indicating the specificity of this assay. Moreover, the aptasensor exhibited high sensitivity with a limit of detection (LOD) at 1 CFU of S. suis and had broad reactivity with S. suis serotypes 1, 1/2, 9, and 14, as well as with S. suis isolated from clinical specimens. Thus, this aptasensor demonstrates proof-of-concept feasibility including clinical sample testing before practical implementation. It holds promise as a practical tool for the early screening and outbreak management of S. suis in a variety of settings, such as clinical laboratories, food safety, and the environment.
Tuberculosis (TB) remains a major global health challenge, with an estimated 10.8 million new cases and 1.25 million deaths in 2023. Despite advances in molecular detection of Mycobacterium TB (MTB), significant diagnostic gaps remain: in 2023, only 48% of newly diagnosed TB cases received rapid diagnostic testing, far below the 100% target. These challenges are intensified in high-burden settings, where sputum collection and distinguishing TB from other illnesses are difficult. The Xpert MTB Host Response (Xpert-HR) assay measures host immune gene expression from blood, shows promise but variable accuracy across studies. Hence, this study will perform an Individual Patient Data Meta-Analysis (IPDMA) to evaluate the diagnostic accuracy, subgroup performance, predictive values and clinical benefit of Xpert-HR compared with conventional sputum-based testing. This systematic review and IPDMA will follow Preferred Reporting Items for Systematic Reviews and Meta-Analyses for Diagnostic Test Accuracy guidelines. Prospective studies including adolescents (>12 years) or adults with presumed TB tested using the Xpert MTB Host-Response assay will be identified through PubMed, Embase and Web of Science. Study quality will be assessed using an adapted diagnostic accuracy tool. Diagnostic accuracy will be pooled using random-effects models, with subgroup analyses where applicable. Decision curve analysis will evaluate clinical utility. Predictive values will be estimated across TB prevalences of 1-10%. Both one-stage and two-stage IPDMA approaches will be explored, and the proportion of unevaluable samples will be reported. The review will be based on deidentified individual patient data to be obtained upon request from the corresponding authors of studies fulfilling all the data sharing agreement. Ethical approval has been obtained from the Ethical Committee of the Medical Faculty of Heidelberg University (Approval No. S-043/2026). The results will be disseminated through publication in a peer-reviewed journal, and through presentations at academic conferences. CRD420251071857.
Human Immunodeficiency Virus, the retrovirus that causes Acquired Immune Deficiency Syndrome, is a major global public health threat. This chronic viral infection diminishes the immune system by attacking CD4 cells. The principal treatment is antiretroviral medication (ART), which significantly increases the life expectancy of HIV patients. However, ART does not address psychological issues, including depression, anxiety, and stress. Psychosocial factors are known to influence HIV disease progression through activation of stress-related biological pathways, including the hypothalamic-pituitary-adrenal (HPA) axis, inflammatory cytokine responses, and monoamine neurotransmitter dysregulation. Mind-body practices such as yoga may modulate these pathways by reducing physiological stress, improving emotional regulation, and enhancing overall well-being. The current trial aims to assess the effectiveness of yoga as an adjunct therapy on psychological parameters (depression, anxiety, and stress), quality of life, and medication adherence of people living with HIV on antiretroviral therapy at a tertiary care hospital in AIIMS, New Delhi, India. This study is a two-arm, parallel-group, open-label, blinded-endpoint, single-center, randomized controlled trial investigating the effects of a yoga therapy as an adjunct therapy in people living with HIV (PLHIV). Participants (n = 192) will be randomized to either 12 weeks of a Yoga therapy program (n = 96) or an Active control group, i.e., a prescribed brisk walk (n = 96). Both groups will receive standard treatment. The primary outcome is anxiety and depression scores (HADS-A and HADS-D), and the secondary outcomes are Stress (PSS), quality of life (WHOQOL-HIV BREF and SF-36 QoL), and medication adherence. The findings of this RCT will help shed light on yoga intervention to address the psychosocial dimensions of HIV. If shown to be effective, yoga as an adjunct intervention may promote a transition in HIV care from a predominantly biomedical framework to a holistic, patient-centered approach encompassing mental health and overall well-being. The study is approved by Institute Research Board Ethics (AIIMSA2969/03.01.2025, RP-46/25, OP-16/02.05.25, OP-18/05.12.2025) and is registered at Clinicaltrials.gov (CTRI/2025/03/081645). CTRI Link- https://www.ctri.nic.in/Clinicaltrials/pmaindet2.php?EncHid=MTIyNjUx&Enc=&userName=HIV,%20Yoga.
In India, no publicly funded seasonal influenza immunization is ongoing, partly because cost-effectiveness is yet to be established. We estimate the cost-effectiveness and budgetary impact of introducing seasonal influenza vaccination among adults aged ≥ 60 years in India, a known high-risk group. Estimates of disease burden including disability adjusted life years (DALYs) were generated from 3-year community cohorts and hospital-based study set up at four sites to estimate incidence of symptomatic acute respiratory infections (ARI), health care utilization, costs and outcomes and applied to India's 2021 estimated population. We used a decision analysis model from an abridged societal perspective for implementation of an inactivated trivalent influenza vaccination programme for older adults (aged ≥ 60 years) using either facility-based or outreach-based approaches. We estimated incremental cost-effectiveness ratio (ICER) per DALY averted and used India's per capita gross domestic product of US$ 2238 for 2021 as the cost-effectiveness threshold. We performed deterministic and probabilistic sensitivity analyses. We also estimated ICER for immunizing all those above 65 years or with comorbidities. Using financial cost of the vaccination programme and the direct medical cost of disease averted we estimated its budgetary impact. In 2021, older adults in India had 5.3 (95% CI: 3.6-7.7) million influenza-ARIs resulting in 36,149 (95% CI: 30,076-43,268) hospitalizations and 84,613 (95% CI: 39,895-155,454) deaths. It amounted to a total of 974,019 DALYs and US$ 66.6 million. As compared with no vaccination, facility-based and outreach-based influenza vaccination had an ICER of US$1979 and US$1851, respectively, below the cost-effectiveness threshold. In all sensitivity and scenario analyses, ICER estimates were below the threshold and vaccinating those above 60 years with specified comorbidities using a passive approach was found to be most cost-effective with least budgetary impact. Annually vaccinating older adults against influenza was found to be cost-effective in the majority of scenarios considered in the study. Given the budgetary implication, it might be prudent to focus on those with co-morbidities.
Invasive fungal infection (IFI) represents a significant global health challenge, particularly in paediatric populations, due to high mortality and severe long-term sequelae. This study aimed to characterize the epidemiological patterns and quantify the disease burden of IFI among hospitalised children in China. Data were obtained from the face sheets of discharge medical records collected between 2016 and 2022 from 30 tertiary children's hospitals, aggregated into the FUTang Updating medical REcords (FUTURE) database. Sociodemographic variables, disease spectrum, pathogen distribution, potential risk factors, length of stay (LOS), and overall disease burden among children with IFI were systematically analysed. A total of 1250 IFI cases were identified, revealing an upward trend in incidence since 2019. The pathogen distribution among the 485 episodes with available microbiological data included Candida (25.36%; 123/485), Aspergillus (23.71%; 115/485), Cryptococcus (22.06%; 107/485), Pneumocystis (21.44%; 104/485), Mucor (4.12%; 20/485), Talaromyces (1.24%; 6/485), Histoplasma (0.82%; 4/485), Blastomyces (0.62%; 3/485), and Sporotrichum (0.62%; 3/485). Disseminated infections accounted for 5.76% (72/1250) of all IFI. Among the 1178 non-disseminated IFI cases, pneumonia (73.34%, 864/1178), central nervous system (CNS) infections (15.53%, 183/1178), and bloodstream infections (8.66%, 102/1178) were the predominant disease types. The most prevalent risk factors were haematological malignancies and myelosuppression. Overall, 181 patients died during their hospitalisation, representing a mortality rate of 14.48%. The incidence of IFI among hospitalised children in tertiary centres in China has risen since 2019, with Candida, Aspergillus, and Cryptococcus identified as the predominant pathogens. These infections are associated with considerable mortality (14.48%). The findings highlight the urgent need for enhanced surveillance, earlier diagnosis, and targeted therapeutic strategies to reduce morbidity and mortality in this high-risk population.
Objective: To explore risk factors for colorectal adenoma (CRA) in non-smoking women, develop a simplified and efficient predictive model, and evaluate its performances with existing risk evaluation tools and in different time periods. Methods: Clinical data were collected from non-smoking women between November 2021 and April 2023. The positive case group included patients with colorectal polyps confirmed as CRA by pathology. Candidate variables were identified through single-factor logistic analysis (P<0.2), and prelimiarily screened using multivariate logistic regression. To obtain the optimal model, a stepwise regression method based on the AIC was employed for predictor selection, and the final prediction model was constructed with the selected predictors. A nomogram and model list were developed to visually demonstrate the contribution of each factor. The model's discrimination and calibration were evaluated and compared with existing risk assessment tools, including the Asia-Pacific Colorectal Screening (APCS) score, its modified version (MAPCS), and the "Colorectal Cancer Screening and Early Diagnosis and Treatment Program (2024 Edition)" of China. A temporal external test set was used to further evaluate the model's stability and predictive performance in a real-world clinical setting. Results: After analyzing data from 1 155 non-smoking women, the final model based on age (5 age groups) and BMI (≥24.0 kg/m²) as the main predictive factors was constructed. The model achieved area under the curve (AUC) values of 0.705 (95%CI: 0.672-0.738) in the training set (n=927) and 0.695 (95%CI: 0.629-0.762) in the validation set (n=228), with calibration curves and Hosmer-Lemeshow tests showing good fitness (P>0.05). A risk threshold of 0.400 was applied, with predicted probabilities ≥0.400 indicating high-risk and <0.400 indicating non-high-risk. The model achieved stable performance in the temporal external test set (n=272) with an AUC of 0.783 (95%CI: 0.730-0.836), further confirming the model's temporal stability and clinical utility. Compared with the existing risk evaluation tools, the values of the model in terms of discrimination are slightly higher than those of the high-risk groups in APCS, MAPCS and "Colorectal Cancer Screening and Early Diagnosis and Treatment Program (2024 Edition)", and the values in terms of specificity and accuracy are also higher. Conclusions: The simplified prediction model based on age and BMI can effectively evaluate CRA risk in non-smoking women, demonstrating high discriminatory power and temporal stability. It can provide more precise risk stratification guidance for early CRA screening with improved efficiency. 目的： 分析不吸烟女性结直肠腺瘤（CRA）的危险因素，构建简洁高效的预测模型，并评估其与现有风险评估工具的比较优势和在不同时间段人群中的效能。 方法： 收集2021年11月至2023年4月不吸烟女性资料，阳性病例组病理结果证实为CRA者。通过单因素logistic回归分析确定候选变量（P<0.2），采用多因素logistic回归分析初步筛选自变量；为得到最优模型采用最小赤池信息准则的逐步回归法进行预测变量筛选，最终筛选出预测变量构建预测模型。绘制nomogram图及建立列表，评估模型区分度及校准度，并与亚太地区结直肠肿瘤筛查评估（APCS）、其修订版（MAPCS）及《结直肠癌筛查与早诊早治方案（2024年版）》等现有风险评估工具进行比较。同时采用时间外部测试集进一步评估模型在实际临床环境中的稳定性和预测性能。 结果： 分析1 155名不吸烟女性的相关数据后，最终模型纳入年龄（分5个年龄段））和BMI（≥24.0 kg/m²）2个主要预测因素。模型在训练集（n=927）和验证集（n=228）的曲线下面积（AUC）分别为0.705（95%CI：0.672~0.738）和0.695（95%CI：0.629~0.762），校准曲线和Hosmer-Lemeshow检验均显示良好拟合性（P>0.05）。以0.400为风险阈值，预测概率≥0.400为高危组，<0.400为非高危组。模型在时间外部验证集（n=272）中表现同样稳定，AUC为0.783（95%CI：0.730~0.836），提示模型具有时间稳定性和临床实用价值。与现有风险评估工具相比，所构建模型在区分度方面的数值稍高于APCS、MAPCS和《结直肠癌筛查与早诊早治方案（2024年版）》等的高危组，且在特异度和准确度方面数值较高。 结论： 基于年龄和BMI的预测模型能有效评估不吸烟女性CRA发病风险，具有较高的区分能力和时间稳定性，可为CRA早期筛查提供更精准的风险分层指导，提高筛查效率。.
This protocol details an optimized method for the production of small stable spheroids, their culture, and 3D imaging, for the study of the endothelial and insulin-producing β cells interactions in a 3D model of pancreatic islets. The 150-200 µm spheroids, mirroring the lowest range of islet sizes, were prepared from a selected ratio combining 1 intra-islet endothelial cells (MS-1 cells) to 20 insulin-secreting cells (β-TC-6). Staining, clearing, and mounting challenges of small spheroids and their tackling by employing low-melting point agarose and the CUBIC clearing technique are detailed, as well as key points for an efficient analysis of the 3D structure with different probes. Data indicate that NTPDASE-ectonucleotidase 3 does not colocalize with insulin in the spheroid model, suggesting varying maturity and functional levels of β-TC6 and that the complete procedure can also be applied to isolated pancreatic islets, with clear probing of intra-islet vessels. These findings underscore the effectiveness of the 3D imaging protocol in revealing complex pancreatic cell organization and interactions within the islet model.
Although next-generation RNA sequencing (RNA-seq) is increasingly incorporated into germline cancer predisposition testing, its diagnostic utility is often limited by low expression of many clinically relevant genes. To improve RNA yield and transcript representation for targeted RNA-seq, we optimized a simple protocol based on short-term lymphocyte culture prepared directly from whole blood collected in Li-heparin tubes. We systematically evaluated biological reproducibility and pre-analytical sample handling variability and demonstrated that whole blood can be stored at 4 °C for up to 5 days prior to lymphocyte cultivation without compromising RNA quality/gene expression. Gene expression was comparable for RNA isolated from K2EDTA and Tempus tubes, whereas short-term lymphocyte culture resulted in a substantial increase in expression of clinically important genes including BRCA1, BRCA2, RAD51C, RAD51D, PALB2, CHEK2, and multiple Fanconi anaemia genes otherwise low expressed in whole blood. Cultivation for 3-5 days did not significantly affect lymphocyte gene expression, providing flexibility for routine dia-gnostics. The protocol also enables inhibition of nonsense-mediated decay to facilitate analysis of variants causing premature termination. As a proof of principle, we characterized the splicing impact of FANCA c.2602-3C>G variant (located in intron 27) using cultured lymphocytes from its carrier. The variant causes deletion of six nucleotides in the mature transcript (ΔE28p(-6)/r.2602_2607del), resulting in an in-frame deletion (p. Gln869_Phe870del). This spliceogenic effect was reliably detectable only in cultured lymphocytes preferentially expressing the full-length FANCA transcript. Overall, short-term lymphocyte culture re-presents a simple and flexible RNA source that enhances variant interpretation in clinical RNA-seq analyses.
Dengue fever is one of the most common mosquito-borne viral infections, with severe cases characterized by plasma leakage, hemorrhage, and multi-organ involvement. Identification of dengue serotypes and reliable biomarkers is essential for predicting disease progression and guiding timely interventions. This prospective cohort study was conducted at a super-speciality tertiary care hospital in southern India from July 2024 to July 2025. A total of 69 patients presenting with dengue warning signs were included in the study. Patients were categorized into the severe dengue group (n = 25) and non severe dengue group (n = 44). Clinical data, laboratory findings, dengue serotype, and serial serum samples collected on Days 1, 4, and 8 were analyzed to evaluate the predictive and monitoring efficacy of Interleukin-6 (IL-6) and Interleukin-8 (IL-8), and followed up till discharge. Out of 69 dengue patients with warning signs, 32 dengue-positive patients were serotyped, which included DEN V-1 (31.3%), DEN V-2 (31.3%), DEN V-3 (15.6%), DEN V-4 (18.8%), and mixed DEN V-(2 + 3) (3.1%). Severe dengue patients exhibited a higher frequency of secondary dengue infection (IgG) than primary dengue infection (88% vs. 12%), with statistically significantly higher packed cell volume, hemoglobin levels, high AST levels, and prolonged activated partial thromboplastin time, as well as lower platelet counts and albumin levels. Platelet transfusion was given to 35 dengue patients, which had also resulted in significant length of stay in hospital in comparison to non-transfused patients. IL-6 and IL-8 levels were significantly elevated in severe dengue patients when compared to non-severe dengue patients on Day 1 and Day 4, followed by a decline on Day 8, corresponding with clinical recovery. However, the elevated IL-8 levels were observed to be significantly associated with longer hospital stays, indicating its potential role as an early predictor of disease progression. The observed co-circulation of multiple serotypes reflects the hyper-endemic pattern reported across India. Early measurement of these cytokines IL-6 and IL-8 helps distinguish severe from non-severe dengue among patients presenting with warning signs. IL-6 and IL-8 may have potential as biomarkers for disease severity. However their role in guiding platelet transfusion requires further investigation in non-severe cases and prioritizing timely management for those at higher risk of severe disease.
Diabetic foot disease is a major public health problem, with an annual National Health Service (NHS) expenditure exceeding £1 billion. Infection increases the risk of major amputation fivefold. Due to the polymicrobial nature of diabetic foot infections, it is often difficult to correctly and rapidly isolate pathological organisms with conventional culture techniques and deliver appropriate narrow-spectrum antimicrobials. Rapid DNA-based technology, using multi-channel arrays, offers a quicker alternative and has previously been used effectively in other settings. We undertook a prospective cohort study of deep tissue samples taken from diabetic foot ulcers (DFUs), comparing samples processed by conventional culture and real-time PCR TaqMan array card (TAC). Fifty samples were taken from 39 patients. The ulcers were of variable chronicity prior to sampling (range: 1-113 weeks) and were sited on the heel (3), midfoot (6) and forefoot (41). TAC results were available an average of 4.3 days earlier than culture results. Seventeen samples had the same organisms detected on culture and TAC. Sixteen of these 17 had additional organisms detected by TAC. The most frequent organisms detected by TAC that were not detected by culture were staphylococci, Enterobacter spp., Pseudomonas spp. and fungi. TAC rapidly and accurately detects clinically relevant organisms from DFUs, providing earlier results than standard culture. This may enable earlier rationalization of antimicrobials and infection control interventions.