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.

Primary small cell carcinoma of esophagus(PSCCE)is a rare and highly aggressive neuroendocrine tumor,accounting for 0.05%to 3.1%of all esophageal malignancies.Its biological characteristics are similar to those of small cell lung cancer,with highly aggressive behavior and early dissemination tendency.It often metasta-sizes rapidly through lymphatic and hematogenous pathways.The prognosis is extremely poor,with a 5-year overall survival rate of less than 15%.There are no large-scale randomized controlled trials,and no standard treatment strategies have been established.In recent years,the treatment of limited-stage PSCCE has become a focal point of research.In traditional treatment paradigms,endoscopic therapy is feasible for very early-stage cases,while radical surgery serves as the primary approach for relatively early-stage patients.For locally advanced cases,two predominant treatment modalities are commonly employed in clinical practice:a surgery-based comprehensive treatment regimen and a radical chemoradiotherapy-centered therapeutic protocol,with no definitive conclusion yet reached regarding the optimal treatment strategy.Concurrently,emerging therapeutic strategies such as immuno-therapy and molecularly targeted therapy have demonstrated remarkable clinical efficacy,thereby providing novel therapeutic opportunities for limited-stage PSCCE.This article aims to review the recent advances in the treatment of limited-stage PSCCE,summarize the current diagnostic and therapeutic landscape,and outline future directions in this field.

AIM:This study aims to investigate the mechanism by which LINC00973 regulates multidrug re-sistance of non-small-cell lung cancer(NSCLC).METHODS:The GEPIA database was employed to analyze the expres-sion levels of LINC00973 in NSCLC and its correlation with patient prognosis in clinical settings.RT-qPCR was utilized to assess LINC00973 expression in various NSCLC cell lines and chemoresistant cells.The migration,invasion,stemness ca-pacity,and drug sensitivity of NSCLC cells with either overexpression or knockdown of LINC00973 were evaluated using wound healing assay,Transwell assay,sphere formation assay,and CCK-8 assay.RNA sequencing was conducted on LINC00973-overexpressing and parental NSCLC cells to identify dysregulated pathways and targets.The LncBase Pre-dicted v.2 and TargetScan databases were used to predict the microRNAs(miRNAs)co-bound by LINC00973 and their target genes.Mimics and inhibitors of candidate miRNAs were synthesized and transfected into NSCLC cells subjected to LINC00973 overexpression or knockdown.Changes in the expression level of LINC00973,and the mRNA and protein ex-pression levels of its target genes were assessed using RT-qPCR and Western blot.RESULTS:Analysis of the GEPIA da-tabase revealed that LINC00973 was significantly up-regulated in lung adenocarcinoma and lung squamous cell carcino-ma,correlating with poor prognosis of NSCLC patients.The LINC00973 expression was elevated in various NSCLC and chemoresistant cell lines(A549/DDP and A549/5-FU).Overexpression of LINC00973 in A549 and H520 cells markedly enhanced their migration,invasion,and stemness capabilities,while concurrently reducing sensitivity to chemotherapy and targeted therapies.RNA sequencing,along with RT-qPCR results,indicated that ATP-binding cassette transporter G5(ABCG5)was activated in LINC00973-overexpressing A549 and H520 cells.Further database analyses and dual trans-fection experiments confirmed that LINC00973 regulated ABCG5 expression through competitive binding with hsa-miR-150-5p.CONCLUSION:LINC00973,which is aberrantly up-regulated in NSCLC specimens and associated with poor clinical prognosis,promotes the expression of ABCG5 through competitive binding with hsa-miR-150-5p.This interaction leads to en-hanced invasion,stemness,and multidrug resistance in NSCLC cells.

This study aims to screen the diagnostic biomarkers for fecal antigen of Echinococcus granulosus(E.granulosus)in dogs with high specificity and sensitivity.The sheep-derived EgPSC artificially infected dogs were collected,and the negative and positive fecal samples of dogs with E.granulosus were prepared by arecoline hydrobromide leakage method.Polyclonal antibody,negative fecal antigen-polyclonal antibody conjugates and positive fecal antigen-polyclonal antibody conju-gates were purified by ammonium sulfate precipitation and affinity chromatography,three groups of samples were detected by ELISA and Western blot,LC-MS/MS and bioinformatics analysis were performed on the three groups of samples.The positive fecal antigen-polyclonal antibody con-jugate was used as the treatment group,the polyclonal antibody and the negative fecal antigen-polyclonal antibody conjugates were used as the control groups to screen the unique peptides of the treatment group.ELISA and Western blot showed that only the positive fecal antigen-polyclonal antibody conjugates were positive.According to LC-MS/MS and bioinformatics analysis,11 unique peptides were screened out only in the treatment group.Among them,3 proteins were related to E.granulosus,namely dysferlin,integrator complex 9 and diagnostic antigen gp50,which were mem-brane-associated proteins,INT complex components and diagnostic antigens.This study has pre-liminarily screened out three candidate canine E.granulosus fecal antigen diagnostic markers,pro-viding a reference for further exploration of diagnostic standards for E.granulosus,screening of echinococcosis target genes,and vaccine development.

Objective To investigate effect of cassava RS3 resistant starch(Ce-RS3)on serum homocysteine(Hcy)level in patients with atherosclerotic cerebral infarction(ACI)during the recovery period.Methods A total of 55 patients with ACI at the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine from October 2023 to October 2024 were selected as subjects.They were devieded into observation group(n=28)and control group(n=27)using a random number table.The control group received atorvastatin calcium,phospholipids,and aspirin,while the observation group received atorvastatin calcium,phospholipids,aspirin,and Ce-RS3.After 12 weeks of treatment,homocysteine(Hcy)levels,carotid plaque diameter,National Institutes of Health stroke scale(NIHSS)scores,Barthel index(BI)scores,and traditional Chinese medicine(TCM)syndrome scores were compared between two groups.Results After treatment,the serum Hcy levels decreased and carotid plaque size reduced in both groups,with the NIHSS scores and TCM syndrome scores also decreased,and observation group was lower than control group(P＜0.05).After treatment,the BI score increased,with observation group higher than control group(P＜0.05).Conclusion The use of Ce-RS3 in the recovery phase of patients with ACI can effectively improve neurological function and enhance treatment efficacy.

Objective:To investigate the mechanism through which ginsenoside Re enhances lipid metabolism and mitochondrial function in H9C2 cardiomyocytes induced by high glucose and high fat, specifically by modulating AMP-activated protein kinase(AMPK)and specificity protein 1(Sp1).Methods:The H9C2 cardiomyocyte model was cultured in a high-glucose and high-lipid(HGHL)environment.Six groups were established: a control group(CON group), a model group(HGHL group), a ginsenoside Re 60 μmol/L group(Re 60 μmol/L), a model+ ginsenoside Re 20 μmol/L group(HGHL+ Re 20 μmol/L), a model+ ginsenoside Re 40 μmol/L group(HGHL+ Re 40 μmol/L), and a model+ ginsenoside Re 60 μmol/L group(HGHL+ Re 60 μmol/L).Cell morphology was assessed using hematoxylin and eosin(HE)staining, while intracellular oil and triglyceride content were evaluated using oil red O staining.Reactive oxygen species(ROS)levels were measured with a fluorescence kit, apoptosis was assessed using TUNEL staining, and the expressions of phosphorylated AMPK, Sp1, nuclear respiratory factor 1(Nrf1), and peroxisome proliferator-activated receptor gamma coactivator 1 alpha(PGC1α)were analyzed via Western blotting.Results:Compared to the control group, the model group exhibited significant increases in cell hypertrophy, ROS levels, apoptosis, and intracellular TG.Conversely, the expressions of pAMPK, Nrf1, and PGC1α were significantly decreased, while the expression of Sp1 was significantly increased( P<0.05).Compared to the model group, treatment with Ginsenoside Re at concentrations of 20 μmol/L, 40 μmol/L, and 60 μmol/L significantly improved cell hypertrophy, reduced ROS levels, alleviated apoptosis, and decreased intracellular TG levels.Additionally, the expressions of pAMPK, Nrf1, and PGC1α were significantly increased, while Sp1 expression was significantly decreased in a dose-dependent manner( P<0.05). Conclusions:Ginsenoside Re enhances lipid metabolism, mitigates mitochondrial oxidative stress, and reduces apoptosis in H9C2 cardiomyocytes induced by high glucose and high lipid conditions.This effect is associated with the regulation of AMPK and Sp1 proteins, leading to increased expressions of pAMPK, Nrf1, and PGC-1α, and decreased expression of Sp1.

This article reports the diagnosis and treatment of a patient with diffuse large B-cell lymphoma (DLBCL) initially manifested as acquired von Willebrand syndrome (AvWS), along with a literature review. The patient, a 22-year-old male, was diagnosed with hereditary von Willebrand disease at the initial stage due to repeated mucosal bleeding, and was later diagnosed with DLBCL (non germinal center type, Ann Arbor stage Ⅳ) due to chest wall mass. Through chemotherapy combined with autologous hematopoietic stem cell transplantation and zebutinib maintenance therapy, the patient achieved sustained complete remission, and the coagulation function returned to normal. This case provides an important reference for the diagnosis and treatment of lymphoma associated AvWS, and highlights the importance of early recognition of basic diseases.

Objective:To explore the safety and effectiveness of laparoscopic treatment for complicated appendicitis in children with and without abscess.Methods:A retrospective analysis was conducted on the clinical data of 363 patients with complicated appendicitis admitted to our department between May 2017 and Sep 2023. Patients were divided into two groups: the group without abscess and the group with abscess, comparing the surgical outcomes and incidence of postoperative complications between two groups.Results:As a result, all 277 cases in the non abscess group completed laparoscopic appendectomy, while all 86 cases in the periappendibular abscess group also completed laparoscopic appendectomy. The history of appendiceal abscess group was longer than that of the group without abscess [5(3-7) d vs. 2(1-3) d, Z=-11.59, P<0.05],the operation time was longer [86 (68-121) min vs. 63 (50-76) min, Z=-7.260, P<0.05], intraoperative bleeding was more [5(3-10) ml vs. 2(2-5) ml, Z=-7.010, P<0.001]. The incidence of postoperative complications was higher in the appendiceal abscess group, with a postoperative abdominal abscess rate of 11.1%,compared to 6.9% ( χ2=1.656, P=0.198), and the incision infection rate of 4.9%,compared to 4.0% ( χ2=0.008, P=0.930), the intestinal obstruction rate was 4.9%,compared to 3.2%, ( χ2=0.158, P=0.691), the intestinal injury rate was 1.3% ,compared to 0,( P=0.226). Conclusions:Laparoscopic appendectomy for periappendiceal abscess in children was safe and effective, especially for patients with a medical history of less than 7 d.

Objective:From the dual perspectives of time and space,a quantitative evaluation of the policies for special outpatient chronic diseases in 31 cities in China over the years is carried out,so as to provide targeted suggestions for relevant departments in the design of medical insurance policies for chronic disease prevention and control.Methods:Combining the relevant literature and the content of the policy text,the policy consistency index model is constructed and quantitative evaluation is carried out based on the results.Results:Of the 421 policies,176 were"Narrowly covered"(41.81%),113 were"Meets expectations"(26.84%),109 were"Focused"(25.89%),and 23 were"Reasonably complete"(5.46%);while 30 cities were"qualified"or above in policy content(χ1),20 cities were"qualified"or above in policy audience(χ2),26 cities were"qualified"or above in policy inclination(χ3),4 cities were"qualified"or above in policy cohesion(χ4),and 24 cities were"qualified"or above in policy objective(χ5).Conclusion:The policies for special outpatient chronic diseases in 31 cities in China are characterized by rich content,wide audience coverage,and diverse policy linkages.To further enhance the security capabilities,it is recommended that sustained efforts be made to consolidate the foundation by standardizing the scope of disease types,strengthening the management of medical treatment in other places and optimizing the interface with other policies.

Objective:To investigate whether repeated tirofiban injections can continuously inhibit platelet aggregation in the arterial duct and affect its closure in neonatal canines.Methods:Four 24-month-old pregnant beagles underwent cesarean sections in two batches (two dogs per batch) 1-2 days before the expected delivery date at the Xuzhou Medical University Animal Experiment Center. The first litter of 21 neonates served as the control group (receiving 10 ml/kg normal saline) and were randomly divided into 1-h ( n=7, injected immediately after birth), 4-h ( n=7, injected at 0 h and 2 h after birth), and 12-h subgroups ( n=7, injected at 0 h, 2 h, 4 h, 6 h, 8 h, and 10 h after birth). The second litter of 18 neonates served as the experimental group (receiving 10 ml/kg tirofiban) with identical subgroup assignments ( n=6 per subgroup). Echocardiography was performed at 1 h, 4 h, and 12 h after birth to measure arterial duct inner diameter, maximum shunt velocity, and left atrial diameter/aortic root diameter (LA/Ao) ratio. Plasma platelet-derived growth factor (PDGF) was detected by enzyme-linked immunosorbent assay, while platelet membrane glycoprotein Ⅱb-Ⅲa in the arterial duct was assessed by Western blot and immunohistochemistry. Data were analyzed using t-tests, one-way ANOVA, Chi square tests, or Fisher's exact test. Results:No significant bleeding tendency occurred in either group. Two control neonates (one each in the 4-h and 12-h subgroups) died. In both control and experimental 1-h subgroups, all arterial ducts remained open, with no significant differences in ductal diameter, shunt velocity, or LA/Ao between groups (all P>0.05). In the 4-h subgroups, all experimental neonates had patent ducts arteriosus, while two controls exhibited closure; the experimental group had larger ductal diameters [(1.05±0.05) vs. (0.55±0.44) mm, t=－2.75, P<0.05)] and higher LA/Ao ratios (1.31±0.09 vs. 1.14±0.03, t=－4.90, P<0.05), but lower maximum shunt velocities [(107.06±17.47) vs. (153.74±12.78) cm/s, t=4.54, P=0.002). In the 12-h subgroups, all the controls had closed arterial ducts, while four of six experimental neonates exhibited closure, though the difference in closure rate was not statistically significant (6/6 vs. 4/6, Fisher's exact test, P=0.455). Plasma PDGF and glycoprotein Ⅱb-Ⅲa levels did not differ between two 1-h subgroups (all P>0.05). However, the 4-h and 12-h experimental subgroups showed lower PDGF levels [(373.5±13.1) vs. (880.3±80.2) pg/ml, t=10.81; (356.7±35.0) vs. (1 111.2±125.3) pg/ml, t=9.74; both P<0.05] and reduced glycoprotein Ⅱb-Ⅲa expression (0.32±0.07 vs. 0.80±0.23, t=3.29; 0.42±0.07 vs. 0.92±0.26, t=3.24; both P<0.05) compared to controls. Conclusion:Repeated tirofiban injections sustainably inhibit platelet aggregation in the arterial duct of neonatal canines and delay ductal closure, suggesting that intraductal platelet aggregation may be one factor influencing this process.