Insect cells of Spodoptera frugiperda support WSSV gene replication but not progeny virion assembly.

The lacking of stable and susceptible cell lines has hampered research on pathogenic mechanism of crustacean white spot syndrome virus (WSSV). To look for the suitable cell line which can sustain WSSV infection, we performed the studies on WSSV infection in the Spodoptera frugiperda (Sf9) insect cells. In consistent with our previous study in vitro in crayfish hematopoietic tissue cells, the WSSV envelope was detached from nucleocapsid around 2 hpi in Sf9 cells, which was accompanied with the cytoplasmic transport of nucleocapsid toward the cell nucleus within 3 hpi. Furthermore, the expression profile of both gene and protein of WSSV was determined in Sf9 cells after viral infection, in which a viral immediate early gene IE1 and an envelope protein VP28 exhibited gradually increased presence from 3 to 24 hpi. Similarly, the significant increase of WSSV genome replication was found at 3-48 hpi in Sf9 cells after infection with WSSV, indicating that Sf9 cells supported WSSV genome replication. Unfortunately, no assembled progeny virion was observed at 24 and 48 hpi in Sf9 cell nuclei as determined by transmission electron microscope, suggesting that WSSV progeny could not be assembled in Sf9 cell line as the viral structural proteins could not be transported into cell nuclei. Collectively, these findings provide a cell model for comparative analysis of WSSV infection mechanism with crustacean cells.

Intrarelationships between suboptimal health status and anxiety symptoms: A network analysis.

BACKGROUND: Suboptimal health status is a global public health concern of worldwide academic interest, which is an intermediate health status between health and illness. The purpose of the survey is to investigate the relationship between anxiety statuses and suboptimal health status and to identify the central symptoms and bridge symptoms. METHODS: This study recruited 26,010 participants aged <60 from a cross-sectional study in China in 2022. General Anxiety Disorder-7 (GAD-7) and suboptimal health status short form (SHSQ-9) were used to quantify the levels of anxiety and suboptimal health symptoms, respectively. The network analysis method by the R program was used to judge the central and bridge symptoms. The Network Comparison Test (NCT) was used to investigate the network differences by gender, place of residence, and age in the population. RESULTS: In this survey, the prevalence of anxiety symptoms, SHS, and comorbidities was 50.7 %, 54.8 %, and 38.5 %, respectively. "Decreased responsiveness", "Shortness of breath", "Uncontrollable worry" were the nodes with the highest expected influence. "Irritable", "Exhausted" were the two symptom nodes with the highest expected bridge influence in the network. There were significant differences in network structure among different subgroup networks. LIMITATIONS: Unable to study the causal relationship and dynamic changes among variables. Anxiety and sub-health were self-rated and may be limited by memory bias. CONCLUSIONS: Interventions targeting central symptoms and bridge nodes may be expected to improve suboptimal health status and anxiety in Chinese residents. Researchers can build symptom networks for different populations to capture symptom relationships.

The crustacean DNA virus tegument protein VP26 binds to SNAP29 to inhibit SNARE complex assembly and autophagic degradation.

Autophagy generally functions as a cellular surveillance mechanism to combat invading viruses, but viruses have evolved various strategies to block autophagic degradation and even subvert it to promote viral propagation. White spot syndrome virus (WSSV) is the most highly pathogenic crustacean virus, but little is currently known about whether crustacean viruses such as WSSV can subvert autophagic degradation for escape. Here, we show that even though WSSV proliferation triggers the accumulation of autophagosomes, autophagic degradation is blocked in the crustacean species red claw crayfish. Interestingly, the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex including CqSNAP29, CqVAMP7, and the novel autophagosome SNARE protein CqSyx12 is required for autophagic flux to restrict WSSV replication, as revealed by gene silencing experiments. Simultaneously, the expressed WSSV tegument protein VP26, which likely localizes on autophagic membrane mediated by its transmembrane region, binds the Qb-SNARE domain of CqSNAP29 to competitively inhibit the binding of CqSyx12-Qa-SNARE with CqSNAP29-Qb-SNARE; this in turn disrupts the assembly of the CqSyx12-SNAP29-VAMP7 SNARE complex, which is indispensable for the proposed fusion of autophagosomes and lysosomes. Consequently, the autophagic degradation of WSSV is likely suppressed by the expressed VP26 protein in vivo in crayfish, thus probably protecting WSSV components from degradation via the autophagosome-lysosome pathway, resulting in evasion by WSSV. Collectively, these findings highlight how a DNA virus can subvert autophagic degradation by impairing the assembly of the SNARE complex to achieve evasion, paving the way for understanding host-DNA virus interactions from an evolutionary point of view, from crustaceans to mammals.IMPORTANCEWhite spot syndrome virus (WSSV) is one of the largest animal DNA viruses in terms of its genome size and has caused huge economic losses in the farming of crustaceans such as shrimp and crayfish. Detailed knowledge of WSSV-host interactions is still lacking, particularly regarding viral escape from host immune clearance. Intriguingly, we found that the presence of WSSV-VP26 might inhibit the autophagic degradation of WSSV in vivo in the crustacean species red claw crayfish. Importantly, this study is the first to show that viral protein VP26 functions as a core factor to benefit WSSV escape by disrupting the assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, which is necessary for the proposed fusion of autophagosomes with lysosomes for subsequent degradation. These findings highlight a novel mechanism of DNA virus evasion by blocking SNARE complex assembly and identify viral VP26 as a key candidate for anti-WSSV targeting.

Study on the determinants of health professionals' performance on diabetes management care in China.

BACKGROUND: As the direct providers of diabetes management care in primary health care facilities (PHFs) in China, health professionals' performance on management care of diabetes determines the quality of services and patients' outcomes. This study aims to analyze the key determinants of health professionals' performance on diabetes management care in PHFs in China. METHODS: We conducted a cross-sectional study in 72 PHFs in 6 cities that piloted the contracted family doctor service (CFDS). Self-developed questionnaire was used to measure three kinds of factors (capacity, motivation and opportunity) potentially influencing the performance of health professionals. The performance of diabetes management care in the study was measured as whether health professionals delivered 7 service items required by the National Basic Public Health Service Guideline with a total of 7 points and was divided into three grades of good, medium and bad. The questionnaire is self-administered by all the health professionals involved in the study with the number of 434. The Chi-square tests were used to compare differences of performance on diabetes management care among health professionals with different characteristics. The ordinal logistic regression was used to analyze the determinants on the performance of diabetes management care. RESULTS: Health professionals who got higher score on diabetes knowledge test had odds of better performance on diabetes management care (OR = 1.529, P < 0.001). health professionals with higher degree of self-reported satisfaction on training (OR = 1.224, P < 0.05) and perception of decreasing workload (OR = 3.336, P < 0.01) had odds of better performance on diabetes management care. While health professionals with negative feeling on information system support had odds of worse performance on diabetes management care (OR = 0.664, P < 0.01). CONCLUSIONS: Attention should be paid to the training of health professionals' knowledge on diabetes management capacity. Furthermore, measures to improve training for health professionals could satisfying their needs for self-growth and improve the motivation of health professionals. The information system supporting management care should be improved continuously to improve the health professionals' working opportunities and decrease the workload.

Pendelluft as a predictor of weaning in critically ill patients: An observational cohort study.

Objective: Weaning failure is associated with adverse clinical outcomes. This study aimed to evaluate the accuracy of pendelluft during the spontaneous breathing trials (SBT) as a predictor of weaning outcome of patients with mechanical ventilation. Methods: An observational cohort study included 60 critically ill patients who were eligible for extubation. Pendelluft and electrical activity of the diaphragm (Edi) were monitored at baseline and every 10 minutes for the first 30 min of SBT denoted as T0, T1, T2, and T3. The pendelluft was measured using electrical impedance tomography (EIT), and Edi parameters were collected by Edi catheter. Patients were followed up after extubation and were divided into success group and failure group. Pendelluft, Edi parameters, respiratory parameters, and clinical outcomes such as intensive care units (ICU) stay, mortality, and 28-day ventilator-free days were compared between the two groups. Receiver operating characteristic (ROC) curves were constructed to evaluate the ability of pendelluft to predict weaning outcome. Results: Fifty patients (50/60) were successfully weaned from the machine and 10 (10/60) failed, with weaning failure rate of 16.7%. Respiratory parameters such as rapid shallow breathing index (RSBI), respiratory rate (RR) and Edi parameters such as maximum value of Edi (Edimax), Edi variation between a maximum and minimum(ΔEdi) in the failure group were higher than those in the success group. The ICU stay and the 28-day ventilator-free days in the failure group were significantly longer than those in the success group. The 28-day mortality rate was higher in the failure group. The pendelluft mainly occurred in the early stage of SBT. Ventral pendelluft and total pendelluft in the failure group were higher than those in the success group at T1. Edimax and ΔEdi were positively correlated with pendelluft. The area under ROC curve (AUC) showed moderate predictive ability for ventral pendelluft in predicting weaning failure at T1 (AUC 0.76, 95% CI 0.58-0.94, cut-off value > 3% global tidal variation). Conclusion: Pendelluft is one of the factors leading to weaning failure, which may be related to diaphragm function. Measuring pendelluft volume maybe helpful to predict weaning.

A Novel Seven Gene Signature-Based Prognostic Model to Predict Distant Metastasis of Lymph Node-Negative Triple-Negative Breast Cancer.

BACKGROUND: The prognosis of lymph node-negative triple-negative breast cancer (TNBC) is still worse than that of other subtypes despite adjuvant chemotherapy. Reliable prognostic biomarkers are required to identify lymph node-negative TNBC patients at a high risk of distant metastasis and optimize individual treatment. METHODS: We analyzed the RNA sequencing data of primary tumor tissue and the clinicopathological data of 202 lymph node-negative TNBC patients. The cohort was randomly divided into training and validation sets. Least absolute shrinkage and selection operator Cox regression and multivariate Cox regression were used to construct the prognostic model. RESULTS: A clinical prognostic model, seven-gene signature, and combined model were constructed using the training set and validated using the validation set. The seven-gene signature was established based on the genomic variables associated with distant metastasis after shrinkage correction. The difference in the risk of distant metastasis between the low- and high-risk groups was statistically significant using the seven-gene signature (training set: P < 0.001; validation set: P = 0.039). The combined model showed significance in the training set (P < 0.001) and trended toward significance in the validation set (P = 0.071). The seven-gene signature showed improved prognostic accuracy relative to the clinical signature in the training data (AUC value of 4-year ROC, 0.879 vs. 0.699, P = 0.046). Moreover, the composite clinical and gene signature also showed improved prognostic accuracy relative to the clinical signature (AUC value of 4-year ROC: 0.888 vs. 0.699, P = 0.029; AUC value of 5-year ROC: 0.882 vs. 0.693, P = 0.038). A nomogram model was constructed with the seven-gene signature, patient age, and tumor size. CONCLUSIONS: The proposed signature may improve the risk stratification of lymph node-negative TNBC patients. High-risk lymph node-negative TNBC patients may benefit from treatment escalation.

Prediction models for acute kidney injury in critically ill patients: a protocol for systematic review and critical appraisal.

INTRODUCTION: Acute kidney injury (AKI) has high morbidity and mortality in intensive care units, which can lead to chronic kidney disease, more costs and longer hospital stay. Early identification of AKI is crucial for clinical intervention. Although various risk prediction models have been developed to identify AKI, the overall predictive performance varies widely across studies. Owing to the different disease scenarios and the small number of externally validated cohorts in different prediction models, the stability and applicability of these models for AKI in critically ill patients are controversial. Moreover, there are no current risk-classification tools that are standardised for prediction of AKI in critically ill patients. The purpose of this systematic review is to map and assess prediction models for AKI in critically ill patients based on a comprehensive literature review. METHODS AND ANALYSIS: A systematic review with meta-analysis is designed and will be conducted according to the CHecklist for critical Appraisal and data extraction for systematic Reviews of prediction Modelling Studies (CHARMS). Three databases including PubMed, Cochrane Library and EMBASE from inception through October 2020 will be searched to identify all studies describing development and/or external validation of original multivariable models for predicting AKI in critically ill patients. Random-effects meta-analyses for external validation studies will be performed to estimate the performance of each model. The restricted maximum likelihood estimation and the Hartung-Knapp-Sidik-Jonkman method under a random-effects model will be applied to estimate the summary C statistic and 95% CI. 95% prediction interval integrating the heterogeneity will also be calculated to pool C-statistics to predict a possible range of C-statistics of future validation studies. Two investigators will extract data independently using the CHARMS checklist. Study quality or risk of bias will be assessed using the Prediction Model Risk of Bias Assessment Tool. ETHICS AND DISSEMINATION: Ethical approval and patient informed consent are not required because all information will be abstracted from published literatures. We plan to share our results with clinicians and publish them in a general or critical care medicine peer-reviewed journal. We also plan to present our results at critical care international conferences. OSF REGISTRATION NUMBER: 10.17605/OSF.IO/X25AT.

Horizontal Inequity in Health Care Utilization among the Middle-Aged and Elderly in China.

BACKGROUND: Equitable utilization of health care is a primary goal of the Chinese health system. This study aimed to examine horizontal inequity in health care utilization and identify the factors resulting in inequity among the middle-aged and elderly in China. METHODS: The data were obtained from the China Health and Retirement Longitudinal Study (CHARLS). We employed the concentration index (CI) and horizontal inequity (HI) to measure inequity in health care utilization. Non-linear regression models were used to decompose the CI into the contribution of each factor. RESULTS: The CIs for the probability of and total number of outpatient visits were 0.0642 and 0.0780, respectively, and those for inpatient visits were 0.1418 and 0.1471, respectively. The HIs were also significantly positive. Living standard was the most important contributor. The contribution of health insurance varied between plans. CONCLUSIONS: This study supported the presence of pro-rich inequity in health care utilization for both probability and frequency among the middle-aged and elderly in China. Narrowing the living standard gap and improving the health insurance benefit package for the elderly should help to resolve these inequities.

Whether New Cooperative Mmedical Schemes reduce the economic burden of chronic disease in rural China.

BACKGROUND: The New Cooperative Medical Scheme (NCMS) provides health insurance coverage for rural populations in China. This study aimed to evaluate changes in household catastrophic health expenditure (CHE) due to chronic disease before and after the reimbursement policies for services of chronic disease were implemented to provide additional financial support. METHODS: The study used data from the household surveys conducted in Shandong Province and Ningxia Hui Autonomous Region in 2006 and 2008. The study sample in village-level units was divided into two groups: 36 villages which implemented the NCMS reimbursement policies for chronic diseases as the intervention group, and 72 villages which did not as the control group. Health care expenditure of more than 40% of household's non-food expenditure was defined as a household with CHE (i.e., impoverishment). The conceptual framework was established based on the Andersen socio-behavioral model of health care utilization to explore how the NCMS reimbursement policies impacted health expenditures. A difference-in-difference model was employed to compare the change in the proportion of households incurring CHE due to chronic disease between the two groups over time. RESULTS: The households that participated in the NCMS were less likely to become impoverished (P<0.05). In addition, the households with both male household head and higher income level were protective factors to prevent CHE (P<0.05). Young households with preschool children suffered less from CHE (P<0.05). The effect of the NCMS reimbursement policies for chronic disease on the CHE was negative, yet not statistically significant (p = 0.814). CONCLUSIONS: The NCMS coverage showed financial protection for households with chronic disease. However, the NCMS reimbursement policies should be strengthened in the future.