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 investigate the relationship between the risk occurrence of rectal cancer and the single nucleotide polymorphisms(SNP) of zinc finger protein 6(ZBED6), and to provide the experimental basis for early diagnosis and treatment of rectal cancer. Methods: Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) technology was used to genotype the ZBED6 gene in 109 randomly selected rectal cancer patients and 110 unrelated healthy controls. To evaluate the relationship between alleles, genotypes and the risk of rectal cancer, unconditional Logistic regression analysis was used toORand 95%CI. Results: The SNP rs7552670 of ZBED6 had significant correlation with the risk of rectal cancer. Compared with the population carrying the TT genotype, those carrying the TC genotype had a 2.653 fold increased risk of rectal cancer(TTvsTC:OR=2.635, 95%CI=1.501-4.690). Other SNPs had no significant correlation with the risk of rectal cancer. Conclusion There is an interaction between the polymorphisms of ZBED6(rs7552670) and rectal cancer. The population carried ZBED6 rs7552670 TC genotype had an growing risk of rectal cancer.

Objective To study the accuracy and safety of oral mucosal exfoliated cell specimens used in the bedside rapid detection of MTHFR C677T genotype by using the fluorescent probe method.Methods The outpatients and inpatients with hypertension visited and admitted in the department of cardiovascular medicine of this hospital from January 2019 to September 2020 were selected.The plasma homocysteine (Hcy) level in all patients was detected in the laboratory,a total of 482 hypertensive patients with Hcy≥10 μmol/L were se-lected,and the oral mucosal cells and whole blood sample were collected in all patients,and the genotypes of the above specimens were detected by the oral mucosal exfoliative cell fluorescent probe method and whole blood sample contrast reagents.If the two test results were inconsistent,the "gold standard" Sanger sequen-cing method was used to detect the whole blood sample for the final determination of MTHFR C677T geno-type.The coincidence rate was compared between the two detection methods,and the probability of adverse e-vents during the samples collection was observed and recorded.The accuracy and safety of fluorescence probe method for detecting MTHFR C677T genotype in the patients with oral mucosa exfoliation was evaluated.Re-sults The oral mucosal exfoliated cell samples and whole blood samples from 482 hypertensive patients were successfully collected,and no obvious adverse reactions occurred during the sampling process.The incidence rate of total mutation of MTHFR C677T gene detected by the fluorescence probe method and contrast reagent all were 73.23％ (353/482),the coincidence rate of homozygous wild type (CC type) in MTHFR C677T gene detected by the two methods was 100.00％ (95％CI:97.11-100.00),which of heterozygous mutant type (CT type) was 99.14％ (95％CI:96.91-99.76),which of homozygous mutant type (TT type) was 99.17％(95％CI:95.47-99.85),the total coincident rate of MTHFR C677T genotype was 99.38％ (95％CI:98.19-99.79)and the detection results consistency Kappa value was 0.9902.Conclusion The detection of MTHFR C677T gene mutation in oral mucosal exfoliated cells by fluorescent probe method is simple with less invasion,moreover which is rapid,safe and accurate.

Objective:To summarize the clinical and electrophysiological characteristics of epilepsy patients with fixation-off sensitivity (FOS).Methods:Using "fixation-off sensitivity" and "epilepsy" as search terms, data were retrieved from the Electroencephalogram Monitoring Center Database of Xijing Hospital, Air Force Military Medical University. Information was collected on epilepsy patients with FOS who were seen at the Xijing Hospital Electroencephalogram Monitoring Center from March 2019 to May 2023. A retrospective analysis of video electroencephalograms and clinical information was conducted, along with follow-up. Single factor analysis and multi-factor Logistic regression analysis were used to analyze the risk factors affecting the prognosis of the patients.Results:A total of 78 patients met the inclusion criteria, including 43 females and 35 males; the age at first epilepsy onset was 12.00 (8.00, 15.25) years. Among these 78 patients, 46 were diagnosed with genetic generalized epilepsy, 11 with self-limited focal epilepy, 9 with familial cortical myoclonic tremor with epilepsy, 7 with developmental and epileptic encephalopathy, 3 with symptomatic epilepsy, and 2 with progressive myoclonic epilepsy. The fixation-off induction trial most commonly triggered seizures in epilepsy with eyelid myoclonia (EEM) patients, with eyelid myoclonia being the most common type of seizure. Twenty-five epilepsy patients with FOS showed a positive photoparoxysmal response, of whom 6 experienced photo-convulsive response. Sixty-eight patients completed follow-up with comprehensive clinical data. At the last follow-up, 45 patients had been free from tonic-clonic seizures for at least one year. A history of febrile seizures ( OR=20.559, 95% CI 2.179-193.997, P=0.008) and cognitive decline ( OR=3.752, 95% CI 1.118-12.588, P=0.032) were identified as risk factors for poor prognosis in epilepsy patients with FOS. The age of first epilepsy onset, the number of anti-seizure medications, and the intermittent photo stimulation trial results had no correlation with prognosis. Conclusions:FOS is more common in female epilepsy patients and can be observed in both generalized and focal epilepsy syndromes. EEM patients are prone to seizures during fixation-off induction trial, and the most common type of seizures is eyelid myoclonus seizure. Patients with epilepsy with FOS who have a history of febrile seizures and cognitive impairment tend to have poor prognosis.

[摘 要] 目的：探讨miR-216b-5p对食管癌Eca109细胞顺铂（DDP）耐药性的影响及其作用机制。方法:采用qPCR法检测miR-216b-5p在食管癌细胞TE-1、KYSE-150、Eca109和耐药细胞Eca109/DDP中的表达水平。利用脂质体转染技术分别将miR-216b-5p mimic及mimic NC、自噬相关蛋白5（ATG5）过表达质粒转染到Eca109/DDP细胞中，用CCK-8、EdU法和FCM分别检测转染后细胞的增殖和凋亡；mRFP-eGFP-LC3双荧光标记实验检测mRFP-eGFP-LC3慢病毒感染后各组细胞自噬发生情况，WB法检测自噬相关蛋白LC3、Beclin 1和P62表达。用荧光素酶报告基因实验验证miR-216b-5p与ATG5的靶向关系，WB法检测ATG5的表达。建立裸鼠Eca109/DDP细胞移植瘤模型，观察miR-216b-5p过表达对移植瘤生长的影响。结果：miR-216b-5p在TE-1、KYSE-150、Eca109和Eca109/DDP细胞中均呈低表达（均P<0.05）。过表达miR-216b-5p可显著抑制Eca109/DDP细胞的增殖并诱导凋亡（均P<0.05），减少细胞中自噬小体数量（P<0.05），下调LC3Ⅱ/LC3Ⅰ比值和Beclin 1蛋白水平、上调P62蛋白水平（均P<0.05）。双荧光素酶报告基因实验证实miR-216b-5p靶向并负调控ATG5的表达（P<0.05），过表达ATG5可使miR-216b-5p mimic对Eca109/DDP细胞增殖、自噬的抑制作用和凋亡的诱导作用明显减弱（均P<0.05），自噬相关蛋白P62表达降低、LC3Ⅱ/LC3Ⅰ比值和Beclin 1表达升高（均P<0.05）。荷瘤实验结果表明，miR-216b-5p过表达可显著抑制裸鼠移植瘤的生长（P<0.05）。结论：miR-216b-5p过表达可逆转食管癌Eca109/DDP细胞对DDP的耐药性，其机制可能与靶向负调控ATG5表达并影响细胞自噬有关。

Objective:To compare the differences in lung function between sanitation workers and the general population undergoing routine physical examinations, and to analyze the risk factors for restricted airflow and severity of the condition in sanitation workers.Methods:This study is a large cross-sectional study called "Shanxin Respiratory Health Screening for Ten Thousand People". A total of 1 036 sanitation workers (sanitation group) and 6 701 individuals from the general population undergoing routine physical examinations (control group) were selected as the original study subjects from June 2021 to April 2022 (before matching). Both groups underwent pre-bronchodilator lung function tests, and the differences in lung function characteristics between the two groups were compared. The sanitation group also completed a questionnaire survey. Multivariate and ordinal multinomial logistic regression analysis were used to analyze the risk factors for airflow limitation and its severity.Results:A total of 1 027 individuals from the sanitation group and 999 individuals from the control group were included in the study. There were no significant differences in age, gender, height, weight, and body mass index (BMI) between the two groups (all P>0.05). The rate of airflow restriction was significantly higher in the sanitation group compared to the control group (22.88% vs 8.81%, P<0.001). In the sanitation group, there was no statistically significant difference in a self-assessment test for chronic obstructive pulmonary disease (CAT) scores between individuals with airflow restriction (235 cases) and those without airflow restriction (792 cases) [(1.50±2.50) vs (1.15±2.03) points, P=0.084]. There were no statistically significant differences in forced vital capacity (FVC) as a percentage of predicted value (FVC%pred) between the two groups. However, the sanitation group had significantly lower %pred for forced expiratory volume in one second (FEV 1%pred), FVC/FEV 1 ratio (FEV 1/FVC%pred), forced expiratory flow at 50% of FVC (FEF 50%%pred), forced expiratory flow at 75% of FVC (FEF 75%%pred), and maximal mid-expiratory flow (MMEF%pred) compared to the control group (all P<0.05). The rates of abnormal FEF 50%%pred, FEF 75%%pred, and MMEF%pred were significantly higher in the sanitation group compared to the control group (17.62% vs 10.31%, 17.04% vs 10.01%, 27.26% vs 18.41%, all P<0.001). Small airway parameters and the rate of airflow restriction were significantly higher in past and current smokers of the sanitation group compared to never smokers (all P<0.05). Multifactorial analysis showed that high BMI ( OR=0.929, 95% CI: 0.885-0.974) was a protective factor for airflow restriction, while high smoking index was a risk factor ( OR=1.020, 95% CI: 1.011-1.030). Ordered multinomial logistic regression analysis showed that high BMI ( OR=0.925, 95% CI: 0.882-0.971) was a protective factor for the severity of airflow restriction, while high smoking index ( OR=1.020, 95% CI: 1.011-1.029) was a risk factor for the severity of airflow restriction. Conclusions:The incidences of airflow limitation and small airway abnormalities in sanitation workers are higher than that in general physical examination population. High smoking index and low BMI are independent risk factors for airflow limitation and its severity.

【Objective】 To analyze the characteristics of peripheral blood mononuclear cells (PBMC) collection in healthy children, and explore the factors affecting collection efficiency (CE). 【Methods】 The PBMC data, involving 70 episodes of apheresis from 42 children during January 2017 and June 2020 were retrospectively analyzed All children were collected in Zhujiang Hospital of Southern Medical University. 【Results】 Multiple linear regression analysis showed that mononuclear cells (MNC) in donor collections from healthy children were positively correlated with anticoagulant dosage, lymphocyte count and monocyte count (P<0.05), meanwhile, negatively correlated with age and platelet count. The PBMC CE was negatively correlated with age, platelet count, and processed whole blood volume (P<0.05). CD34⁺ cells (×10⁷ /kg)was negatively correlated with age, meanwhile, positively correlated with numbers of collection and processed whole blood volume(r=-0.79). No statistical differences in red blood cell count, platelet count, lymphocyte count, monocyte count of healthy child donors were notable before versus after apheresis. 【Conclusion】 MNC can be collected effectively in children of different ages. The PBMC collection efficiency was related to age. Meanwhile, the higher the lymphocytes and monocytes were before apheresis, the more MNC were collected. The efficiency of MNC collection would decrease when the apheresis volume of the children exceeded their total blood volume twice. However, the absolute value of CD34+ cells in the final yields would increase.

Objective:To investigate the effects of microRNA(miR)-133b on epithelial-mesenchymal transition(EM)of human small airway epithelial cells induced by cigarette smoke extracts(CSE)and its regulatory mechanisms.Methods:The miR-expression profiles with microarray in airway epithelial cells of patients with chronic obstructive pulmonary disease were searched in the Gene Expression Omnibus(GEO)database, and the differentially expressed miRs were searched and verified by a real-time fluorescence quantitative method(qRT-PCR). Human small airway epithelial cells(HSAEpiC)were divided into the control group, the CSE group, the CSE+ miR-133b inhibitor transfection group(inhibitor group)and the CSE+ miR-133b inhibitor negative control transfection group(inhibitor control group)according to different intervention methods.Levels of miR-133b and mRNA levels of transforming growth factor(TGF)-β1, Smad2, E-cadherin and vimentin were detected by RT-PCR; Protein levels of E-cadherin and vimentin were detected by enzyme-linked immunosorbent assays(ELISA)and Western blotting.Results:Nine differentially expressed miRs were found in GSE53519, with miR-133b showing the most significant differential in thee HSAEpiC cell model after verification.CSE induced morphological changes in HSAEpiC cells, and miR-133b inhibitors could partially reverse the morphological changes in cell mode.mRNA and protein expressions of E-cadherin were decreased and expression of Vimentin mRNA and protein were increared in CSE induced HSAEpiC cells( F=9.09、12.35、7.57、101.87, P=0.015、0.007、0.023、0.000); miR-133b inhibitors partally reversed the mRNA and protein expressions of E-cadherin and Vimentin( F=40.59、27.74、15.87、20.42, P=0.000、0.001、0.004、0.002). CSE induced incresed expression of TGF-β1 mRNA and Smad mRNA in HSAEpiC cells, and miR-133b inhibitors partially reversed the changes in TGF-β1 mRNA and Smad mRNA( F=17.25、64.15, P=0.003、0.000). Conclusions:miR-133b may regulate CSE-related HSAEpiC cell EMT through the TGF-β1/Smad pathway.

Objective To explore the difference in the regulating effect of electroacupuncture on serum insulin (Ins) and fat consumption between male and female simple obesity rat and the possible mechanism of its action.Methods A rat model of simple obesity was made by induction with sodium glutamate. The rats were randomized into model and electroacupuncture groups (male and female), 10 rats each. A normal group of 10 rats (male and female) was established as a control. Points Quchi, Zhongwan, Guanyuan, Housanli, Fenglong and Sanyinjiao were selected in the electroacupuncture group. Stimulation with a low frequency of 2 Hz was provided after needle insertion. The treatment was given once daily for 28 consecutive days. Serum Ins content, and greater omentum, pericardiac and perirenal fat weights were measured in the male and female groups of rats before and after electroacupuncture intervention. The comparisons were made.Results Serum Ins content, and greater omentum, pericardiac and perirenal fat weights were significantly lower in the electroacupuncture male and female groups of rats than in the model male and female groups of rats (P<0.01) and also in the electroacupuncture male group of rats than in the electroacupuncture female group of rats (P<0.05).Conclusions Electroacupuncture has different degrees of weight-reducing effect in both male and female obesity rats. The reducing effect on serum Ins content, and greater omentum, pericardiac and perirenal fat weights is better in male obesity rats.

Objective To investigate early and differential diagnosis of Alzheimer disease and vascular dementia (VD)using FDG-PET scan. Methods Clinical data was collected from AD,VD and normal control(NC). 18F-2-fluo-ro-deoxy-D-glucose(18F-FDG)PET scan was conducted to detect the cerebral FDG metabolism. The average standard uptake value (SUV) of cerebral regions was expressed as semiquantitative index relative to ipsilateral cerebellum. Re-sults There were no differences in age and the mean total scores of the mini mental status examination (MMSE) between patients with VD and those with AD.The AD group showed that the SUV was significantly decreased in the right frontal, parietal,temporal lobe,hippocampus,temporo-parietal junction,and bilateral posterior cingulate cortex compared to the VD and in the bilateral posterior cingulate cortex,temporo-parietal junction,frontal and temporal lobe comparing to the NC(P≤0.01). SUV was significantly increased in the right parietal,posterior cingulate cortex and hippocampus whereas was significantly decreased in the left frontal lobe,temporo-parietal junction and thalamus in the VD compared with NC group(P≤0.01). Conclusion AD patients have typical features of 18F-FDG PET which may be helpful for the diagnosis of AD in the early stage.