Objective To elucidate the kinetic characteristics of viral replication and transcription,an in vitro model of viral replication and transcription was established.Utilizing single-molecule magnetic tweezers technology,the dynamic process of SARS-CoV-2 RNA-dependent RNA polymerase(RdRp)during in vitro replication and transcription was investigated.Methods The force field of the single-molecule magnetic tweezer system was corrected using DNA fragments,followed by the construction of RNA fragments to explore the kinetics of RdRp replication and transcription in vitro.Results The force field calibration results were consistent with the worm-like chain model(WLC).The ssRNA strand was found to be approximately 0.3 pm longer than the dsRNA strand and could be stably extended under a 30 pN force field.The average synthesis rate of RdRp extension was determined to be 3.27 nt/s,with an average processivity of 886 nt.Conclusions By implementing force calibration in single-molecule magnetic tweezers,the real-time tracking of RdRp kinetics during the full-cycle replication-transcription process(initiation,elongation,and termination)in vitro was achieved,thereby constructing a mechanistic model of RdRp-driven nucleic acid synthesis.This study provides a basis for further investigating the kinetics of viral RdRp in physiological processes including replication,transcription,and backtracking under varied in vitro environments using single-molecule magnetic tweezers,and establishes a single-molecule manipulation framework for evaluating the effects of therapeutic compounds on viral replication-transcription processes in vitro.

OBJECTIVE To explore the effect of hydrogen peroxide disinfectant on terminal disinfection in wards of medical institutions.METHODS The surfaces of highly frequent contact objects of the wards of the First Affilia-ted Hospital of Nanchang University from which the public health center patients were discharged between Apr.2024 and Jun.2024 were respectively disinfected with 0.5％(low)and 5％(high)concentrations of hydrogen peroxide disinfecting wipes,totally 180 samples were randomly collected before and after the disinfection,and the pathogens were detected.The air of the wards from which the public health center patients were discharged be-tween Jul.2024 and Aug.2024 were disinfected with hydrogen peroxide dry mist disinfection device,and 90 sam-ples were respectively collected before and after the disinfection.Geobacillus stearothermophilus was used as the biological indicator and placed at various points within the air-disinfected wards to evaluate the disinfection level.The environmental sampling results and distribution of bacteria were observed and compared.RESULTS The qualified rates of disinfection of the object surfaces were 95.56％(86/90)and 98.89％(89/90)respectively for the low and high concentratioins of hydrogen peroxide disinfecting wipes,and there was no significant difference in the disinfection effect.Totally 120 strains of pathogens were isolated from unqualified samples before the disinfection,among which gram-negative bacteria(69.17％)were dominant,and the isolation rate of multidrug-resistant or-ganisms was 22.50％(27/120);only 1 strain of carbapenem-resistant Acinetobacter baumannii was isolated after the disinfection.The qualified rate of disinfection of air in the wards was 96.00％by 7.5％hydrogen peroxide dry mist disinfection device,the average bacterial colony counts in the air were 2 CFU/(5 min·vsl)after the dis-infection,and the killing rate of Geobacillus stearothermophilus was 100.00％by the air disinfection.CONCLUSION The hydrogen peroxide disinfectant can meet the requirement for terminal disinfection of the wards of the medical institutions,and it is portable and highly efficient.

Objective To observe the repair effect and mechanism of Danhuang Powder-containing serum on high glucose-induced vascular endothelial cell injury.Methods Danhuang Powder-containing serum was prepared.Human umbilical vein endothelial cells(HUVECs)were cultured to be divided into control group,recombinant human epidermal growth factor(called"growth factor"for short)group,Danhuang Powder group,high glucose group,high glucose+growth factor group,and high glucose+Danhuang Powder group.After corresponding intervention in each group for 48 hours,the cell ultrastructure and autophagy were observed under transmission electron microscope,apoptosis was detected by flow cytometry,and the protein expression levels of vascular endothelial growth factor(VEGF),epidermal growth factor(EGF)and basic fibroblast growth factor(bFGF)in the cells were detected by Western Blot.Results(1)The intra-mitochondrial ridges in the control group were clearly visible,autophagosomes and autolysosomes were fewer;mitochondria in the high glucose group were swollen and irregular,and appeared vacuolated;and the more typical autophagy-like structures were seen in the high glucose+Danhuang Powder group.(2)Compared with the high glucose group and high glucose+growth factor group,the apoptosis rate of cells in the high glucose+Danhuang Powder group was significantly decreased(P<0.05).(3)Compared with the high glucose group and the high glucose+growth factor group,the protein expression levels of VEGF,EGF and bFGF in the cells of the high glucose+Danhuang Powder group were significantly increased(P<0.05).Conclusion Danhuang Powder-containing serum can reduce the high glucose-induced damage in HUVEC cells,and its mechanism may be related to the activation of mitochondrial autophagy,and the inhibition of apoptosis,as well as the up-regulation of the expression of VEGF,EGF and bFGF.

Xiaohuang Qudan decoction (XHQDD) is a classical traditional Chinese medicine (TCM) formula widely used in the treatment of cholestatic liver injury. Despite its widespread use, the protective mechanism of XHQDD against cholestatic liver injury remains incompletely understood. The aim of this study was to investigate whether XHQDD mediates its beneficial effects by inhibiting the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway and regulating TH17/Treg balance. To this end, the researchers used Sprague-Dawley (SD) rats and established a cholestatic liver injury model by oral administration of alpha-naphthylisothiocyanate (ANIT). The experimental group was divided into six groups: Control (CON), ANIT, ursodeoxycholic acid (UDCA), XHQDD-low dose (XHQDD-L) group, XHQDD-medium dose (XHQDD-M) group, and XHQDD-high dose (XHQDD-H) groups. Then, after 7 d of treatment, various tests were performed to verify the results. Firstly, XHQDD and its drug-containing serum were analyzed by ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS), and 14 blood-entry components were identified. Then, bile flow was monitored and found to be significantly reduced in the model group, which was significantly reversed in the UDCA and XHQDD groups. To further assess ANIT-induced liver injury, hematoxylin and eosin (H&E) and Sirius red staining, alongside transmission electron microscopy (TEM), were employed to observe liver tissues, revealing hepatocellular injury, cholestasis, and hepatic fibrotic changes. Serum inflammatory factors and liver injury indicators were assessed using enzyme-linked immunosorbent assay (ELISA), indicating an inflammatory state in ANIT-induced liver injury rats. The expression levels of JAK2/STAT3-related genes and proteins in liver and intestinal tissues were measured via quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunohistochemistry, immunofluorescence (IF) staining, and Western blottting (WB) assays. These studies revealed that the inflammatory state of liver-injured rats was inextricably linked to the inflammatory cascade associated with the JAK2/STAT3 pathway and that XHQDD may exert anti-inflammatory efficacy by inhibiting the JAK2/STAT3 pathway. Flow cytometry was used to determine the percentage of T helper 17 (Th17)/regulatory T (Treg) cells in serum and hepatocytes, and it was further found that XHQDD was able to regulate Th17/Treg immune homeostasis in liver-injured rats. The findings suggest that XHQDD markedly alleviates inflammation in ANIT rats, potentially treating cholestasis and liver injury through JAK2/STAT3 inhibition and Th17/Treg balance regulation.
Animals ; STAT3 Transcription Factor/immunology* ; Janus Kinase 2/immunology* ; Drugs, Chinese Herbal/pharmacology* ; Rats, Sprague-Dawley ; 1-Naphthylisothiocyanate/adverse effects* ; Male ; Rats ; Th17 Cells/immunology* ; Cholestasis/immunology* ; Signal Transduction/drug effects* ; T-Lymphocytes, Regulatory/immunology* ; Chemical and Drug Induced Liver Injury/immunology* ; Liver/drug effects*

Uterine arteriovenous fistula (UAVF), a relatively rare vascular anomaly, may lead to life-threatening hemorrhage and poses diagnostic and management challenges for primary hospitals during initial encounters. This article reports four cases of UAVF treated with Drospirenone and ethinyl estradiol tablets at Department of Gynaecology, Zhuji Maternal and Child Health Hospital between 2023 and 2024. Combined with literature review, we systematically analyzed the etiological mechanisms, clinical characteristics, and diagnostic-therapeutic strategies of this condition, with particular focus on evaluating the efficacy and safety of conservative pharmacological management. By summarizing case features and individualized treatment protocols, this study aims to provide clinical references for early identification, accurate diagnosis, and rational intervention of UAVF, thereby reducing risks of massive hemorrhage and long-term complications while improving patient prognosis.

Uterine arteriovenous fistula (UAVF), a relatively rare vascular anomaly, may lead to life-threatening hemorrhage and poses diagnostic and management challenges for primary hospitals during initial encounters. This article reports four cases of UAVF treated with Drospirenone and ethinyl estradiol tablets at Department of Gynaecology, Zhuji Maternal and Child Health Hospital between 2023 and 2024. Combined with literature review, we systematically analyzed the etiological mechanisms, clinical characteristics, and diagnostic-therapeutic strategies of this condition, with particular focus on evaluating the efficacy and safety of conservative pharmacological management. By summarizing case features and individualized treatment protocols, this study aims to provide clinical references for early identification, accurate diagnosis, and rational intervention of UAVF, thereby reducing risks of massive hemorrhage and long-term complications while improving patient prognosis.

Objective To elucidate the kinetic characteristics of viral replication and transcription,an in vitro model of viral replication and transcription was established.Utilizing single-molecule magnetic tweezers technology,the dynamic process of SARS-CoV-2 RNA-dependent RNA polymerase(RdRp)during in vitro replication and transcription was investigated.Methods The force field of the single-molecule magnetic tweezer system was corrected using DNA fragments,followed by the construction of RNA fragments to explore the kinetics of RdRp replication and transcription in vitro.Results The force field calibration results were consistent with the worm-like chain model(WLC).The ssRNA strand was found to be approximately 0.3 pm longer than the dsRNA strand and could be stably extended under a 30 pN force field.The average synthesis rate of RdRp extension was determined to be 3.27 nt/s,with an average processivity of 886 nt.Conclusions By implementing force calibration in single-molecule magnetic tweezers,the real-time tracking of RdRp kinetics during the full-cycle replication-transcription process(initiation,elongation,and termination)in vitro was achieved,thereby constructing a mechanistic model of RdRp-driven nucleic acid synthesis.This study provides a basis for further investigating the kinetics of viral RdRp in physiological processes including replication,transcription,and backtracking under varied in vitro environments using single-molecule magnetic tweezers,and establishes a single-molecule manipulation framework for evaluating the effects of therapeutic compounds on viral replication-transcription processes in vitro.

OBJECTIVE To explore the effect of hydrogen peroxide disinfectant on terminal disinfection in wards of medical institutions.METHODS The surfaces of highly frequent contact objects of the wards of the First Affilia-ted Hospital of Nanchang University from which the public health center patients were discharged between Apr.2024 and Jun.2024 were respectively disinfected with 0.5％(low)and 5％(high)concentrations of hydrogen peroxide disinfecting wipes,totally 180 samples were randomly collected before and after the disinfection,and the pathogens were detected.The air of the wards from which the public health center patients were discharged be-tween Jul.2024 and Aug.2024 were disinfected with hydrogen peroxide dry mist disinfection device,and 90 sam-ples were respectively collected before and after the disinfection.Geobacillus stearothermophilus was used as the biological indicator and placed at various points within the air-disinfected wards to evaluate the disinfection level.The environmental sampling results and distribution of bacteria were observed and compared.RESULTS The qualified rates of disinfection of the object surfaces were 95.56％(86/90)and 98.89％(89/90)respectively for the low and high concentratioins of hydrogen peroxide disinfecting wipes,and there was no significant difference in the disinfection effect.Totally 120 strains of pathogens were isolated from unqualified samples before the disinfection,among which gram-negative bacteria(69.17％)were dominant,and the isolation rate of multidrug-resistant or-ganisms was 22.50％(27/120);only 1 strain of carbapenem-resistant Acinetobacter baumannii was isolated after the disinfection.The qualified rate of disinfection of air in the wards was 96.00％by 7.5％hydrogen peroxide dry mist disinfection device,the average bacterial colony counts in the air were 2 CFU/(5 min·vsl)after the dis-infection,and the killing rate of Geobacillus stearothermophilus was 100.00％by the air disinfection.CONCLUSION The hydrogen peroxide disinfectant can meet the requirement for terminal disinfection of the wards of the medical institutions,and it is portable and highly efficient.

Objective:To explore the value of serum matrix metalloproteinase-1 (matrix metalloproteinase-1), and phosphatase and tensin hmmlogydeleted on ten (PTEN), in predicting osteoporosis (OP) in women with gestational diabetes mellitus (GDM) .Methods:The clinical data of.218 pregnant women with GDM treated in Linyi People’s Hospital from Sep. 2019 to Feb. 2023 were retrospectively collected and included in the GDM group. Another 98 healthy pregnant women who underwent physical examination during the same period were selected as the control group. Age, body mass index (BMI), blood glucose related indexes and serum MMP-1 and PTEN levels were compared between the two groups. According to the occurrence of osteoporosis (OP) in the second and third trimester of pregnancy, the GDM group was divided into OP group and non-OP group. Logistic regression analysis was used to explore the influencing factors of OP in pregnant women with GDM in the second and third trimesters. ROC curve was drawn to analyze the predictive value of serum MMP-1 and PTEN levels for osteoporosis in GDM pregnant women in the second and third trimesters.Results:Body mass index (BMI), fasting blood glucose (FBG), glycated hemoglobin A1c (HbA1c) and serum MMP-1 and PTEN levels in GDM group were higher than those in control group, but BUA, SOS and SI were lower than those in control group ( t=6.18, 12.68, 9.91, 11.42, 7.70, 5.63, 5.76, 9.84, all P<0.05). Among 218 GDM women, 29 cases had OP and 189 cases did not have OP. There were no significant differences in fasting blood glucose, glycosylated hemoglobin, blood calcium, blood magnesium, or blood phosphorus between the two groups (statistical value: 0.96, 0.86, 1.35, 1.06, 0.49, P>0.05). Logistic regression analysis showed that age, abnormal increase of MMP-1 and PTEN levels were independent risk factors for OP in GDM women in the second and third trimesters [ OR (95% CI) =1.260 (1.103-1.440) ,1.075 (1.031-1.120) ,1.055 (1.023-1.089), all P<0.05]. ROC curve showed that the AUC of MMP-1 and PTEN in predicting OP in GDM pregnant women in the second and third trimesters were 0.824 and 0.777, respectively. The AUC of the combination of the two in predicting OP in GDM pregnant women in the second and third trimesters was 0.933, and the specificity and sensitivity were 83.04% and 93.48%, respectively, which were higher than those of single detection. Conclusions:MMP-1 and PTEN in early pregnancy may play a potential role in the prediction of OP in pregnant women with GDM. Close monitoring of MMP-1 and PTEN levels may provide new ideas for the clinical diagnosis and treatment of OP high-risk patients.

Objective:To understand the current status of gastroscopy in diagnosing gastric lesions in general population, and to recommend the optimal age for the first gastroscopy and intervals for repeated gastroscopy.Methods:The gastroscopy records of residents aged 18-80 years in Yinzhou District of Ningbo, Zhejiang Province, between April 2010 and December 2021 were analyzed retrospectively. The detections of gastric lesions across different years, age and genders were described. Goodness of fit tests were applied to compare the differences in detection rates of different lesions in first-time endoscopy in different age groups and different populations. Generalized additive models were used to fit the trend of age specific gastric lesion detection rate explore the optimal age for gastroscopy. The appropriate gastroscopy intervals were determined according to the progress of the gastric lesions detected in repeated gastroscopy.Results:A total of 237 751 participants with 344 398 gastroscopy records were included in analyses. A total of 5 597 cases of chronic atrophic gastritis (CAG), 9 796 cases of intestinal metaplasia (IM), 165 cases of low-grade intraepithelial neoplasia (LGIN), 52 cases of high-grade intraepithelial neoplasia (HGIN) and 435 cases of gastric cancer were detected by the first gastroscopy. The overall detection rate of gastric lesions increased significantly in age group 45-70 years, and remained stable after 70 years old, with LGIN and HGIN showing notable increases at 50 and 55 years old, respectively. Repeated gastroscopy detected CAG, IM, LGIN, and HGIN at a higher rate compared with the first gastroscopy. Normal/superficial gastritis progressed in 3-5 years, whereas CAG or more severe lesions progressed in 1-6 years.Conclusion:Gastroscopy is recommended for general population aged 45 years and above. Furthermore, gastroscopy can be performed every 3-5 years for individuals with normal endoscopy results and once a year for patients with CAG or more severe gastric lesions.