The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.

ObjectiveTo evaluate the clinical efficacy and safety of Baoshen prescription in the treatment of stage Ⅳ diabetic nephropathy （DN） in the patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction， and to explore the mechanism of this prescription delaying the disease progression. MethodsA randomized， controlled， double-blind， multicenter clinical trial was conducted， in which 94 stage Ⅳ DN patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction were randomly assigned into Baoshen prescription and control groups （47 cases）. The treatment lasted for 12 weeks. The primary efficacy indicators were mainly renal function indexes， including urine albumin-to-creatinine ratio （UACR）， 24-hour urine total protein （24 h-UTP）， serum creatinine （SCr）， and estimated glomerular filtration rate （eGFR）. The secondary efficacy indicators were metabolic memory of hyperglycemia， podocyte epithelial-to-mesenchymal transdifferentiation-related indexes， and TCM syndrome score. ResultsAfter 12 weeks of treatment， the Baoshen prescription group showed lowered levels of advanced glycation end products （lgAGEs）， connective tissue growth factor （CTGF）， type Ⅳ collagen （Col-Ⅳ）， receptor of AGEs （RAGE）， urinary fibroblast-specific protein-1 （FSP-1）， UACR， 24 h-UTP， and glycated hemoglobin （HbAlc） （P<0.05）， and an upward trend of miR-21 mRNA. The control group showed elevated levels of SCr and UREA and lowered levels of urinary FSP-1， eGFR， and HbAlc （P<0.05）. After treatment， the Baoshen prescription group had lower levels of lgAGEs， CTGF， urinary FSP-1， SCr， UACR， and 24 h-UTP and higher levels of Col-Ⅳ and eGFR than the control group （P<0.05）. In addition， the Baoshen prescription group showed statistically significant differences in SCr， eGFR， UACR， and 24 h-UTP before and after treatment （P<0.05）. ConclusionBaoshen prescription can effectively improve the renal function， reduce the urinary protein level， and alleviate clinical symptoms in stage Ⅳ DN patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction. The mechanism may be related to the metabolic memory of hyperglycemia and epithelial-to-mesenchymal transdifferentiation of podocytes.

Objective To explore the application effects of an innovative teaching model(blended 5MP)that organically combines blended teaching and the five-minute teaching method in clinical teaching for operating room nursing students.Methods Using convenience sampling,94 nursing students who interned in operating room of Kunming Children's Hospital from July 2023 to March 2024 were randomly assigned to observation and control groups,with 47 students in each group.For the control group,a traditional blended learning approach was utilized,whereas the observation group implemented a blended 5MP teaching model.Results The observation group's post-class assignment scores,theoretical exam scores,and practical skills assessment scores were all significantly higher than those in the control group,with a statistically significant difference(P<0.05).The observation group also had higher scores in the Self-Assessment of Clinical Reasoning and Reflection Scale,with a statistically significant difference(P<0.05).Furthermore,the observation group reported higher satisfaction rates regarding teaching methods,teaching formats,theoretical instruction,practical instruction,and overall satisfaction compared to the control group,with statistically significant differences(P<0.05).Conclusion The blended approach combines online and offline instructional modalities,allowing for a more strategic allocation of educational resources.This methodology enables students to thoroughly acquire essential knowledge and skills while simultaneously fostering advanced clinical reasoning and reflective competencies.By adopting this innovative educational model,institutions can significantly enhance the academic experience and professional development of nursing students.

AIM:To investigate the preventive and therapeutic effects of sesamin(SES)on fatty liver disease in rats with hypertension combined with dyslipidemia,and to explore the potential mecha-nisms based on mRNA-seq.METHODS:Spontane-ously hypertensive rats(SHRs)were fed a high-fat,high-cholesterol diet to establish a rat model of hy-pertension combined with dyslipidemia,and then treated with SES for 16 weeks continuously.The ex-periment was divided into four groups:WKY,SHR,Model,and Model+SES(160 mg·kg-1·d-1).Blood pressure was measured using the tail-cuff method.Body weight was monitored,and body mass index was calculated.Liver morphology was detected by ultrasound,and liver thickness was measured.Liver wet weight was weighed,and liver index was calcu-lated.Liver volume was detected by the water dis-placement method.Serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),aspartate amino-transferase(AST),and total bile acids(TBA)were de-tected by ELISA.Liver sequencing analysis was per-formed using mRNA-seq.Liver histomorphological changes were observed by HE staining.The degree of hepatic steatosis was observed by Oil Red O stain-ing,and the degree of hepatic fibrosis was observed by MASSON staining.The mRNA expression of Al-dh1a7,Nnmt,Irs2,Pltp,and Scd was detected by q-PCR.The protein expression of Scd,Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was detected by Western blotting.RESULTS:After 16 weeks of continuous SES administration to rats with hypertension combined with dyslipidemia,blood pressure was significantly reduced(P＜0.01),and body weight was decreased.Serum TG,TC,and LDL-C levels were decreased,while HDL-C levels were increased.Serum ALT and AST levels were decreased.Liver weight,organ in-dex,liver thickness,and liver volume were de-creased.The degree of hepatic steatosis and hepat-ic fibrosis was improved.A total of 545 differentially expressed mRNAs were identified in the livers of rats in each group,of which 278 were upregulated and 267 were downregulated.Among the 27 com-monly differentially expressed mRNAs,five mRNAs related to lipid metabolism were screened,namely Aldh1a7,Nnmt,Irs2,Pltp,and Scd.KEGG enrich-ment analysis showed that the enriched pathways were AMPK and PPAR.Further validation revealed that in the SES-treated group,the mRNA expression of Scd in the liver was decreased,while the mRNA expression of Nnmt was increased.The protein ex-pression of Scd was decreased,while the protein ex-pression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was increased.CONCLUSION:SES has preven-tive and therapeutic effects on fatty liver disease in rats with hypertension combined with dyslipidemia,and its mechanism of action may be related to the reduction of Scd expression levels in the liver and the increase in the expression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ.

Objective:To explore the relationships among psychological resilience,spiritual health,and illness uncertainty in cancer patients,and to analyze the mediating role of disease uncertainty.Methods:The cancer patients were selected by convenience sampling method from Feb 2024 to May 2024 in the Department of Oncology of a Grade Ⅲ-A general hospital in Wuhu City.Data were collected using a general information questionnaire,the Mishel Uncertainty in Illness Scale(MUIS),the Functional Assessment of Chronic Illness Therapy-Spiritual Well-being Scale(FACIT-SP-12,Chinese version),and the Connor-Davidson Resilience Scale(CD-RISC-10).Pearson correlation analysis was conducted to assess the relationships among psychological resilience,illness uncertainty,and spiritual health.The mediating effect of illness uncertainty was tested using Hayes'PROCESS Model 4 and the Bootstrap method.Results:The total scores of spiritual health,psychological resilience and illness uncertainty of cancer patients was(25.11±7.19),(24.36±6.75)and(67.75±13.06),respectively.The spiritual health was positively correlated with psychological resilience(r=0.415,P＜0.01)and negatively correlated with illness uncertainty(r=-0.398,P＜0.01).The psychological resilience was negatively correlated with illness uncertainty(r=-0.668,P＜0.01).Illness uncertainty partially mediated the relationship between psychological resilience and spiritual health,accounting for 35.29%of the total effect.Conclusions:The spiritual health of cancer patients is at a moderate level.Enhancing psychological resilience and reducing illness uncertainty can alleviate psychological burden and improve spiritual health,thereby promoting overall quality of life.

Sj?gren′s syndrome (SS) is a prevalent systemic autoimmune disorder characterized by significant heterogeneity and complexity, which present considerable challenges in diagnosis and management. In 2017, the British Society for Rheumatology (BSR) released guidelines for the management of adult SS. Based on this, the BSR identified 19 key issues and subsequently formulated 47 evidence-based recommendations in 2024,including 11 recommendations supported by level A evidence. Compared to the previous guidelines, the 2024 version has expanded its scope to include novel recommendations in diagnostic approaches (such as salivary gland ultrasonography and labial salivary gland biopsy)and in the management of specific patient populations (such as patients with juvenile-onset and pregnant patients). There are also substantial updates regarding the local treatment of keratoconjunctivitis sicca. However, the use of systemic medications still requires additional robust evidence from clinical studies. This article aims to critically discuss these updates to provide rheumatologists with valuable insights for clinical decision-making and to offer guidance for patients with SS in their self-management strategies.

Objective To develop a Treatment Burden Scale for patients with chronic heart failure(CHF)and evaluate its reliability and validity,aiming to provide an effective tool for assessing treatment burden levels in this population.Methods According to Cumulative Complexity Model,the study was conducted in 2 phases.A preliminary item pool was established through literature review and semi-structured interviews,from August 2022 to March 2023,followed by expert consultation to finalize the initial scale.A convenience sample of CHF patients from 4 tertiary A hospitals in Hunan and Hubei provinces was conducted to refine scale items and assess psychometric properties,from June 2023 to April 2024.Results The final scale contained 26 items.Exploratory factor analysis revealed 6 domains,including family financial support burden,healthcare utilization burden,the challenges patients face in acquiring knowledge about proper medication use and water/sodium dietary management,the behavioral burden associated with tracking daily water/sodium intake and symptom patterns,psychological burden,and medication management burden,cumulatively explaining 68.661％of variance.Confirmatory factor analysis demonstrated satisfactory model fit;the x2/df was 2.076;the root mean square error of approximation was 0.070;the normed fit index was 0.912;the content validity score was 0.974.The total Cronbach's α coefficient of the scale was 0.903;the split-half reliability was 0.785;the test-retest reliability was 0.936.Conclusion The Treatment Burden Scale for CHF patients developed in this study has good reliability and validity psychometric properties,and it can be used to evaluate treatment burden levels and influencing factors in clinical practice.

Objective:To analyze the distinctive urinary exosomal metabolites in patients with lupus nephritis (LN).Methods:In this case-control study, urine samples were collected from 29 LN patients (5 males and 24 females; age, 32?±?12 years old) admitted to the Department of Rheumatology in the Nanjing University Medical School Affiliated Drum Tower Hospital form October 20th, 2022 to March 10th, 2024, as well as from 20 healthy volunteers (4 males and 16 females; age 34?±?11 years old) undergoing routine physical examinations. Exosomes were isolated using polyethylene glycol precipitation, and metabolomic profiling was performed using liquid chromatography-mass spectrometry (LC-MS). The preprocessed data matrix was analyzed via principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) use R lamguage. Differential metabolites were screened based on the variable importance in projection (VIP) values derived from the OPLS-DA model and Student′s t-test. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis.Results:Metabolomic analysis identified a total of 963 metabolites. Based on the criteria of VIP>1 and P<0.05, three differential exosomal metabolites—L-tryptophan, L-isoleucine, and L-phenylalanine—were ultimately selected. The areas under the ROC curves were 0.774, 0.783, and 0.789, respectively, with a combined AUC of 0.847. Conclusion:This study preliminarily identified characteristic urinary exosomal metabolites in LN patients, providing novel insights and strategies for the subsequent identification of biomarkers in LN.

OBJECTIVE To explore the functional substance basis of Jinhong Tablet in the treatment of chronic superficial gastri-tis(CSG).METHODS Three different models were constructed to investigate the anti-inflammatory and antioxidant effects,func-tional material basis of Jinhong Tablet:inflammatory model in human gastric epithelial cells(GES-1)induced by lipopolysaccharide(LPS),LPS-induced inflammatory model in mouse gastric organoids,and ethanol-induced oxidative damage model in GES-1 cells.MTS assay was performed to detect cell proliferation activity;qPCR was applied to measure the relative mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-8(IL-8)in cells and gastric organoids;and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and reactive oxygen species(ROS)in cells were detected.RESULTS Jinhong Tablet and 10 functional components significantly reduced the relative expression of inflammation-relat-ed genes TNF-α,IL-1β,IL-6,and IL-8 in LPS-induced GES-1 cells and gastric organoids,suggesting that these 10 components are the functional substance basis for the anti-inflammatory effects of Jin Hong Tablet.Jin Hong Tablet and 11 functional components markedly decreased the levels of MDA and ROS and increased the activity of SOD,indicating that these 11 components were the func-tional substance basis of the antioxidant effects of Jinhong Tablet.CONCLUSION Through in vitro cell and gastric organoid experi-ments,it has been preliminarily determined that allocryptopine,corydaline,dehydrocorydaline,palmatine hydrochloride,chlorogenic acid,costunolide,rutin,quercitrin,dehydrocostus lactone,tetrahydrocoptisine,isochlorogenic acid B,toosendanin,protopine,and quercetin are the functional material basis of Jinhong Tablet in treating CSG,accumulating scientific evidence for the enhancement of the quality standards of Jinhong Tablet.