OBJECTIVE: Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro. METHODS: The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining. RESULTS: The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism. CONCLUSION Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

【Objective】 To analyze the effects of different fortified feeding methods on nutritional metabolism and growth rate of preterm very low birth weight infants (VLBWI), in order to provide new clues for improving the prognosis of the preterm infants. 【Methods】 A total of 115 cases of premature VLBWI admitted to Department of Neonatology, The First Affiliated Hospital of Kunming Medical University from January 2019 to December 2020 were included in this study, and were divided into fortified breastfeeding group (HFM group), mixed feeding group, and premature formula feeding group (PF group) based on their feeding methods. The effects of different feeding methods on the nutritional metabolism and growth rate of premature VLBWI were analyzed. 【Results】 1) The hospitalization time of infants in the HFM group was shorter than that in PF group and mixed feeding group (t=7.185, 6.924, P<0.05). 2) The proportion of necrotizing enterocolitis (NEC) in the HFM group during hospitalization was lower than that in the PF group (P<0.05); the proportions of late onset septicemia(LOS) and extra uterine growth restriction(EUGR) in the HFM group during hospitalization were lower than those in the PF group (χ²=5.030, 4.147, P<0.05); the proportion of LOS was lower than that of the mixed feeding group(χ²=6.589, P<0.05). 3) During hospitalization, the proportions of abdominal distension, bloody stools and increased eosinophils in the HFM group were lower than those in the PF group (P<0.05), which in mixed feeding group was lower than those in PF group (Fisher exact test, P<0.05). 4) At discharge, the weight and length growth rate of the HFM group were higher than those of the mixed feeding group (t=3.722, 0.425, P<0.001) and the PF group (t =6.015, 0.496, P< 0.001). 【Conclusion】 Fortified breastfeeding can more effectively increase the growth rate of VLBWI in premature infants, improve nutritional metabolism, reduce complications and adverse feeding reactions related to premature infants, and is safer and more effective.

Mismatch repair-deficient/microsatellite instability-high(dMMR/MSI-H)gastric cancer represents a distinct molecular subtype of tumors characterized by pronounced sensitivity to immune checkpoint inhibitors(ICIs)attributed to its unique immune microenvironment and elevated mutation burden.Various clinical studies underscore the efficacy of ICIs in treating dMMR/MSI-H gastric cancer;however,chal-lenges such as primary and acquired resistance persist.Overcoming resistance and identifying optimal ICIs for its treatment remain critical clinical issues.This review delineates the mechanisms of ICIs,recent advances in their therapeutic application for dMMR/MSI-H gastric can-cer,and ongoing challenges in combating resistance,aiming to guide clinical practice effectively.

Objective To explore the value of non-contrast CT findings of acute ischemic stroke(AIS)for predicting early prognosis after mechanical thrombectomy.Methods Data of 161 AIS patients from clinical center 1 who underwent mechanical thrombectomy were retrospectively analyzed.The patients were divided into training set(n=113)and internal test set(n=48)at the ratio of 7∶3,while 79 AIS patients who underwent mechanical thrombectomy from clinical center 2 were retrospectively enrolled as external test set.According to the National Institutes of Health stroke scale(NIHSS)scores 7 days after thrombectomy,patients'prognosis were classified as good(＜15 points)or poor(≥15 points).Pre-treatment non-contrast CT images of patients were reviewed,and CT findings were comparatively analyzed.Independent predictors of patients'early prognosis after mechanical thrombectomy were obtained with sequential univariate and multivariate logistic regressions,and a predicting model was established and visualized as a nomogram.The receiver operating characteristic curve was drawn,and the distinction was assessed with the area under the curve(AUC),then calibration was assessed with Hosmer-Lemeshow goodness of fit test,and the net benefit was evaluated with decision curve analysis(DCA).Results Alberta stroke program early CT score(ASPECTS),hyperdense middle cerebral artery sign(HMCAS)and basal ganglia calcification were all independent predictors of early prognosis of AIS after mechanical thrombectomy(all P＜0.05).The predictive model was established combining the above 3 variables and then visualized as a nomogram to predict prognosis of AIS after mechanical thrombectomy,with AUC of 0.776 in internal test set(χ2=6.052,P=0.417)and 0.800 in external test set(χ2=2.269,P=0.811).DCA showed that the nomogram might provide clinical net benefit within certain threshold probability ranges.Conclusion ASPECTS,HMCAS and basal ganglia calcification were all independent predictors of early prognosis of AIS after mechanical thrombectomy.The nomogram originated from predicting model combining the three could be used to somewhat accurately predict poor early prognosis after mechanical thrombectomy.

Objective To explore the effect and safety of right unilateral modified electroconvulsive therapy(RUL-MECT)for major depressive disorder patients(MDD).Methods A randomized controlled trial was conducted on 70 patients with MDD who were randomly divided into a study group and a control group.The study group underwent age-based RUL-MECT,while the control group underwent bitemporal MECT.The participants were evaluated using the 17-item Hamilton depression scale(HAMD-17),MATRICS consensus cognitive battery(MCCB)and orientation recovery tests(ORT).Any adverse reactions that occurred during each intervention process were recorded.Results Before treatment,there were no significant differences in the HAMD-17(32.89±5.68 vs.33.54±4.78)between the two groups(P>0.05).The HAMD-17 score of the intervention group was 6.83±4.68,while the control group was 7.20±4.60 after 8 interventions,repeated measures analysis of variance showed the time effect(P<0.001)was significant.The intergroup effect and interaction effect was not significant(P>0.05).In terms of MCCB scores,there were significant main effect(P<0.001)in connectivity tests,symbol coding,language memory,spatial breadth,number sequence,maze test,visual memory,emotional management and the duration of continuous operation.The intergroup effects of language memory,number sequence,visual memory,speech fluency,and continuous operation were significant(P<0.05).The interaction effect of language memory and continuous operation were significant(P<0.05).After the intervention,the recovery of orientation time was significantly shorter in the study group than that in the control group[(508.57±104.48)s vs.(631.66±212.27)s](P<0.05).The incidence of adverse reactions between two groups(28.6% vs.40.0% )has no significance(P>0.05).Conclusions Compared with bitemporal MECT,RUL-MECT has comparable efficacy in treating depressant and better performance in improving cognitive function and recovery of orientation.

Objective To investigate the relationship between docosahexaenoic acid (DHA) level and the onset of attention deficit hyperactivity disorder (ADHD) based on Mendelian Randomization (MR).Meth-ods The instrumental variables for DHA and ADHD were extracted from the genome-wide association stud-ies (GWAS) data.To ensure the stablity of the study results,the ADHD datasets were selected from two dif-ferent studies.The The dataset for DHA levels included 115,006 individuals,the ADHD1 dataset consisted of 55374 individuals,including 20183 cases and 35191 controls,the ADHD2 dataset consisted of 225534 indi-viduals,including 38691 cases and 186843 controls,the comprehensive MR and sensitivity analyses were per-formed.The MR analyses were performed by the inverse variance weighted (IVW) method,MR-Egger regression analysis and the weighted median method (WM),utilizing the odds ratio (OR) and effect coefficient (β) as evaluation indictors to explore the bidirectional causal relationship between DHA and ADHD.Results There exists a causal link between DHA levels and the onset of ADHD (ADHD1 IVW:OR=0.833,95％CI:0.719-0.964,P=0.014;ADHD2 IVW:OR=0.895,95％CI:0.808-0.992,P=0.034).Additionally,a reverse causal relation-ship between ADHD and DHA levels (IVW:β=-0.073,95％CI:-0.103 to-0.043,P<0.001) was found.Conclusion The potential bidirectional causal relationship exists between DHA level and ADHD.

ObjectivesTo investigate characteristics and influencing factors of short-term variation of systolic blood pressure of community hypertension patients in Shanghai. MethodsBased on the standardized blood pressure measurement data of hypertensive patients in Shanghai （2018‒2021） and the noninfectious chronic disease management system database， coefficient of variation of systolic blood pressure （CV_S） was described and the influencing factors were analyzed. ResultsAmong 112 680 community hypertension patients， males accounted for 46.87% with a median age of 69 years. CV_S was 0.038 6 ± 0.035 0. Generalized linear model analysis showed the following influencing factors of CV_S： gender， females were higher than males， B=0.032， P<0.001； age group， 60‒69 years， 70‒79 years old group， and ≥80 years groups were all higher than <50 years group， B=0.042， 0.056， and 0.074 respectively with P values of 0.020， 0.002， and <0.001 respectively； smoking， smoking cessation and smoking everyday was lower than never smoking， B=-0.032 and-0.028 respectively with P values of 0.023 and 0.007 respectively； systolic blood pressure， 140‒159 mmHg group， 160‒179 mmHg group， and ≥180 mmHg group were lower then <140 mmHg group， B=-0.039， -0.091， and -0.175 respectively with P values all <0.001； and measurement season， autumn was lower than spring， （B=-0.056， P<0.001）. The paired test showed that CV_S calculated from the three measurements was 1.04% higher than that calculated from the two measurements （P<0.001）. ConclusionThe coefficient of variation of systolic blood pressure of community hypertension patients in Shanghai has large variation， and was influenced by many factors.

The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR associated proteins) system is an adaptive immune system of bacteria and archaea against phages, plasmids and other exogenous genetic materials. The system uses a special RNA (CRISPR RNA, crRNA) guided endonuclease to cut the exogenous genetic materials complementary to crRNA, thus blocking the infection of exogenous nucleic acid. According to the composition of the effector complex, CRISPR-Cas system can be divided into two categories: class 1 (including type Ⅰ, Ⅳ, and Ⅲ) and class 2 (including type Ⅱ, Ⅴ, and Ⅵ). Several CRISPR-Cas systems have been found to have very strong ability to specifically target RNA editing, such as type Ⅵ CRISPR-Cas13 system and type Ⅲ CRISPR-Cas7-11 system. Recently, several systems have been widely used in the field of RNA editing, making them a powerful tool for gene editing. Understanding the composition, structure, molecular mechanism and potential application of RNA-targeting CRISPR-Cas systems will facilitate the mechanistic research of this system and provide new ideas for developing gene editing tools.
CRISPR-Cas Systems/genetics* ; RNA/genetics* ; Bacteria/genetics* ; Gene Editing ; Archaea

Background/Aims: The accuracy of endosonographers in diagnosing gastric subepithelial lesions (SELs) using endoscopic ultrasonography (EUS) is influenced by experience and subjectivity. Artificial intelligence (AI) has achieved remarkable development in this field. This study aimed to develop an AI-based EUS diagnostic model for the diagnosis of SELs, and evaluated its efficacy with external validation. Methods: We developed the EUS-AI model with ResNeSt50 using EUS images from two hospitals to predict the histopathology of the gastric SELs originating from muscularis propria. The diagnostic performance of the model was also validated using EUS images obtained from four other hospitals. Results: A total of 2,057 images from 367 patients (375 SELs) were chosen to build the models, and 914 images from 106 patients (108 SELs) were chosen for external validation. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the model for differentiating gastrointestinal stromal tumors (GISTs) and non-GISTs in the external validation sets by images were 82.01%, 68.22%, 86.77%, 59.86%, and 78.12%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in the external validation set by tumors were 83.75%, 71.43%, 89.33%, 60.61%, and 80.56%, respectively. The EUS-AI model showed better performance (especially specificity) than some endosonographers.The model helped improve the sensitivity, specificity, and accuracy of certain endosonographers. Conclusions We developed an EUS-AI model to classify gastric SELs originating from muscularis propria into GISTs and non-GISTs with good accuracy. The model may help improve the diagnostic performance of endosonographers. Further work is required to develop a multi-modal EUS-AI system.

Ribosomes are abundant,large RNA-protein complexes that are the sites of all protein synthesis in cells.Defects in ribosomal proteins(RPs),including proteoforms arising from genetic variations,alternative splicing of RNA transcripts,post-translational modifications and alterations of protein expression level,have been linked to a diverse range of diseases,including cancer and aging.Comprehensive character-ization of ribosomal proteoforms is challenging but important for the discovery of potential disease biomarkers or protein targets.In the present work,using E.coli 70S RPs as an example,we first developed a top-down proteomics approach on a Waters Synapt G2 Si mass spectrometry(MS)system,and then applied it to the HeLa 80S ribosome.The results were complemented by a bottom-up approach.In total,50 out of 55 RPs were identified using the top-down approach.Among these,more than 30 RPs were found to have their N-terminal methionine removed.Additional modifications such as methylation,acetylation,and hydroxylation were also observed,and the modification sites were identified by bottom-up MS.In a HeLa 80S ribosomal sample,we identified 98 ribosomal proteoforms,among which multiple truncated 80S ribosomal proteoforms were observed,the type of information which is often overlooked by bottom-up experiments.Although their relevance to diseases is not yet known,the integration of top-down and bottom-up proteomics approaches paves the way for the discovery of proteoform-specific disease biomarkers or targets.