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.

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 evaluate the diagnostic efficacy of FibroScan combined with various noninvasive diagnostic models for liver fibrosis in patients with chronic hepatitis B (CHB) complicated with nonalcoholic fatty liver disease (NAFLD), and to establish a new predictive model with common clinical indicators.Methods:From January 2016 to May 2024, the clinical data of 118 CHB patients complicated with NAFLD from the First Affiliated Hospital of Zhengzhou University, who underwent liver biopsy and FibroScan examination were respectively analyzed. According to the Scheuer scoring system, different diagnostic endpoints were set based on the degree of liver fibrosis (S0 to S1, ≥S2, ≥S3, and S4), fibrosis stage ≥S2 was designated as the criterion for significant liver fibrosis. Fibrosis-4 (FIB-4), γ-glutamyl transpeptidase (GGT) to platelet ratio (GPR), GGT-age-platelet-international normalized ratio (GAPI) model, S index, King index and Forns index were calculated according to the common clinical indicators. The independent t test or Mann-Whitney U test was used to compare the two groups. Spearman rank correlation was used to analyze the correlation between each noninvasive diagnostic method and the degree of liver fibrosis. Receiver operating characteristic curve (ROC) was plotted, and the DeLong test was performed to compare the area under the curve (AUC), and to evaluate the predictive value of FibroScan combined with various noninvasive diagnostic models for the diagnosis of liver fibrosis. The laboratory indicators were compared between patients with non-significant liver fibrosis and patients with significant liver fibrosis. And the indicators with statistically significant differences ( P<0.05) in the univariate analysis were further analyzed by multivariate logistic regression to establish a new predictive model for liver fibrosis. Hosmer-Lemeshow test was used to assess the model′s goodness of fit. Results:The results of Spearman rank correlation showed that FIB-4, GPR, FibroScan, GAPI model, S index, King index, and Forns index were positively correlated with the stage of liver fibrosis ( r=0.413, 0.458, 0.512, 0.473, 0.533, 0.380 and 0.478, all P<0.001). The results of ROC analysis indicated that among combination of FibroScan and other diagnostic models, the AUC values of FibroScan+ FIB-4, FibroScan+ Forns index were relatively high in ≥S2 and ≥S3, which were 0.804 and 0.907, respectively. The platelet count ((200.65±50.89)×10 9/L vs. (169.96±63.68)×10 9/L), total cholesterol ((4.69±0.77) mmol/L vs. (4.32±1.00) mmol/L), high-density lipoprotein (HDL) (1.28 (1.05, 1.46) mmol/L vs. 1.08 (0.92, 1.21) mmol/L), total protein (74.00 (70.63, 77.08) g/L vs. 68.80 (64.60, 73.55) g/L), albumin (47.06 (44.65, 48.81) g/L vs. 44.70 (41.55, 46.20) g/L), and globulin (26.80 (24.48, 29.70) g/L vs. 25.80 (23.05, 27.60) g/L) of the non-significant liver fibrosis group were higher than those of the significant liver fibrosis group, and the differences were statistically significant ( t=2.74, 2.09; Z=-3.30, -3.88, -3.95, -2.01; P=0.007, =0.040, =0.001, <0.001, <0.001, =0.044). GGT (27.50 (17.00, 41.75) U/L vs. 37.00 (22.50, 87.00) U/L), the liver stiffness measurement (LSM) in the non-significant hepatic fibrosis group was lower than the significant liver fibrosis group (6.85 (5.60, 9.26) kPa vs. 11.60 (7.08, 17.26) kPa), and the differences were statistically significant ( Z=-2.73, -4.39; P=0.006, <0.001). The result of multivariate logistic regression analysis revealed that globulin, albumin, HDL, and LSM were independent factors of liver fibrosis ( OR (95% confidence interval)=0.865 (0.759 to 0.985), 0.804 (0.691 to 0.935), 0.128 (0.023 to 0.711), and 1.251 (1.091 to 1.433), respectively; P=0.029, 0.025, 0.019, 0.001). A novel model, GLAH, was established with globulin, LSM, albumin, and HDL. The AUC for diagnosing liver fibrosis degree ≥S2, ≥S3, and S4 was 0.847, 0.938, and 0.909, respectively, which were higher than those of the above models. The positive predictive value for diagnosing liver fibrosis degree ≥S2 with GLAH>1.12 as the cutoff value was 95.8%. The negative predictive value for excluding fibrosis stage ≥S2 with GLAH<-1.41 was 92.3%. This approach could reduce the number of liver biopsies by 48.3% (57/118), with an accuracy of 94.7% (54/57). Conclusions:The clinical value of FibroScan combined with FIB-4 or Forns index is better in the diagnisis of fibrosis stage ≥S2 and ≥S3. The GLAH model has higher diagnostic value and can accurately predict the degree of liver fibrosis in CHB patients complicated with NAFLD, thus reducing the need for liver biopsy.

Objective To verify the diagnostic efficiency and application value of fecal syndecan 2(SDC2)gene methylation detection in intestinal tumor screening.Methods The clinical data of 1 456 patients with fecal SDC2 gene methylation detection in this hospital from November 2021 to December 2023 were ana-lyze retrospectively.The detection positive rate,colonoscopic compliance,sensitivity,specificity,positive pre-dictive rate and negative predictive rate were analyzed.The pathological diagnosis served as the gold standard.The receiver operating characteristic(ROC)curve and the area under the curve(AUC)were used to judge the diagnostic effect.Results In the results in 1 456 cases of fecal SDC2 gene methylation detection,90 cases were positive with a positive rate of 6.2％.The positive rate had no statistical difference between different sexes(P＞0.05).The positive rate in the patients ≥50 years old was higher than that in the patients＜50 year old(P＜0.05).Among 90 cases of detection results positive,67 cases completed the enteroscopic examination and the enteroscopic compliance rate was 74.4％.The enteroscopic compliance rate had no statistical difference be-tween the different sexes and among different ages of patients(P＞0.05).Among 67 cases of enteroscopic ex-amination completion,there were 6 cases(9.0％)of colorectal cancer,17 cases(25.4％)of progressive stage adenoma,15 cases(22.4％)of non-progressive stage adenoma,6 cases(9.0％)of non-adenomatous polyp and the lesion detection rate was 65.7％.Among 112 cases of fecal SDC2 gene methylation detection negative,there were 2 cases(1.8％)of progressive stage adenoma and 22 cases(19.6％)of non-progressive stage ade-noma.The sensitivity and specificity of this detection for colorectal cancer and progressive stage adenoma were 92.0％and 71.4％,respectively,which had obvious diagnostic significance for colorectal tumor(AUC=0.721,P＜0.001).Conclusion The fecal SDC2 gene methylation detection has an important clinical value in the preliminary screening of colorectal cancer.

OBJECTIVE To observe the efficacy of cetrorelix combined with aspirin in preventing early-onset ovarian hyperstimulation syndrome （OHSS）. METHODS A retrospective analysis was conducted on clinical data from 38 early-onset OHSS patients， who received treatment in our hospital from January 1st to July 1st， 2022. These patients were divided into intervention group （19 cases） and control group （19 cases） according to the therapy regimen. On the first day after oocyte retrieval surgery， the control group was given aspirin enteric-coated tablets 100 mg orally until menstruation began. The intervention group was given cetrorelix for injection 0.25 mg subcutaneously， for consecutive 3 days+aspirin enteric-coated tablets （same usage and dosage as the control group）. The first luteal phase， the degree of OHSS， and the ovarian volume， ascites volume， serum estradiol （E2）， white blood cell count （WBC）， hematocrit （HCT）， neutrophil ratio （NEUT%）， D-dimer （DD）， prothrombin time （PT）， fibrinogen （Fib） after oocyte retrieval surgery were observed and measured in 2 groups. RESULTS The first luteal phase was significantly shorter， and the proportions of median and severe OHSS cases were significantly lower in the intervention group compared to the control group （P＜0.05 or P＜0.01）. After oocyte retrieval surgery， the intervention group showed significantly lower ovarian volume， ascites volume， serum E2， WBC， NEUT%， HCT， DD and Fib compared to the control group， but PT of intervention group was signiticantly higher than that of control group （P＜0.05）. CONCLUSIONS Cetrorelix combined with aspirin is more effective in preventing early-onset OHSS than aspirin alone.

Objective:To evaluate the diagnostic efficacy of FibroScan combined with various noninvasive diagnostic models for liver fibrosis in patients with chronic hepatitis B (CHB) complicated with nonalcoholic fatty liver disease (NAFLD), and to establish a new predictive model with common clinical indicators.Methods:From January 2016 to May 2024, the clinical data of 118 CHB patients complicated with NAFLD from the First Affiliated Hospital of Zhengzhou University, who underwent liver biopsy and FibroScan examination were respectively analyzed. According to the Scheuer scoring system, different diagnostic endpoints were set based on the degree of liver fibrosis (S0 to S1, ≥S2, ≥S3, and S4), fibrosis stage ≥S2 was designated as the criterion for significant liver fibrosis. Fibrosis-4 (FIB-4), γ-glutamyl transpeptidase (GGT) to platelet ratio (GPR), GGT-age-platelet-international normalized ratio (GAPI) model, S index, King index and Forns index were calculated according to the common clinical indicators. The independent t test or Mann-Whitney U test was used to compare the two groups. Spearman rank correlation was used to analyze the correlation between each noninvasive diagnostic method and the degree of liver fibrosis. Receiver operating characteristic curve (ROC) was plotted, and the DeLong test was performed to compare the area under the curve (AUC), and to evaluate the predictive value of FibroScan combined with various noninvasive diagnostic models for the diagnosis of liver fibrosis. The laboratory indicators were compared between patients with non-significant liver fibrosis and patients with significant liver fibrosis. And the indicators with statistically significant differences ( P<0.05) in the univariate analysis were further analyzed by multivariate logistic regression to establish a new predictive model for liver fibrosis. Hosmer-Lemeshow test was used to assess the model′s goodness of fit. Results:The results of Spearman rank correlation showed that FIB-4, GPR, FibroScan, GAPI model, S index, King index, and Forns index were positively correlated with the stage of liver fibrosis ( r=0.413, 0.458, 0.512, 0.473, 0.533, 0.380 and 0.478, all P<0.001). The results of ROC analysis indicated that among combination of FibroScan and other diagnostic models, the AUC values of FibroScan+ FIB-4, FibroScan+ Forns index were relatively high in ≥S2 and ≥S3, which were 0.804 and 0.907, respectively. The platelet count ((200.65±50.89)×10 9/L vs. (169.96±63.68)×10 9/L), total cholesterol ((4.69±0.77) mmol/L vs. (4.32±1.00) mmol/L), high-density lipoprotein (HDL) (1.28 (1.05, 1.46) mmol/L vs. 1.08 (0.92, 1.21) mmol/L), total protein (74.00 (70.63, 77.08) g/L vs. 68.80 (64.60, 73.55) g/L), albumin (47.06 (44.65, 48.81) g/L vs. 44.70 (41.55, 46.20) g/L), and globulin (26.80 (24.48, 29.70) g/L vs. 25.80 (23.05, 27.60) g/L) of the non-significant liver fibrosis group were higher than those of the significant liver fibrosis group, and the differences were statistically significant ( t=2.74, 2.09; Z=-3.30, -3.88, -3.95, -2.01; P=0.007, =0.040, =0.001, <0.001, <0.001, =0.044). GGT (27.50 (17.00, 41.75) U/L vs. 37.00 (22.50, 87.00) U/L), the liver stiffness measurement (LSM) in the non-significant hepatic fibrosis group was lower than the significant liver fibrosis group (6.85 (5.60, 9.26) kPa vs. 11.60 (7.08, 17.26) kPa), and the differences were statistically significant ( Z=-2.73, -4.39; P=0.006, <0.001). The result of multivariate logistic regression analysis revealed that globulin, albumin, HDL, and LSM were independent factors of liver fibrosis ( OR (95% confidence interval)=0.865 (0.759 to 0.985), 0.804 (0.691 to 0.935), 0.128 (0.023 to 0.711), and 1.251 (1.091 to 1.433), respectively; P=0.029, 0.025, 0.019, 0.001). A novel model, GLAH, was established with globulin, LSM, albumin, and HDL. The AUC for diagnosing liver fibrosis degree ≥S2, ≥S3, and S4 was 0.847, 0.938, and 0.909, respectively, which were higher than those of the above models. The positive predictive value for diagnosing liver fibrosis degree ≥S2 with GLAH>1.12 as the cutoff value was 95.8%. The negative predictive value for excluding fibrosis stage ≥S2 with GLAH<-1.41 was 92.3%. This approach could reduce the number of liver biopsies by 48.3% (57/118), with an accuracy of 94.7% (54/57). Conclusions:The clinical value of FibroScan combined with FIB-4 or Forns index is better in the diagnisis of fibrosis stage ≥S2 and ≥S3. The GLAH model has higher diagnostic value and can accurately predict the degree of liver fibrosis in CHB patients complicated with NAFLD, thus reducing the need for liver biopsy.

Objective:To explore the multidisciplinary management that centred on gastroenterology department, and follow-up study of children with Alagille syndrome(ALGS).Methods:The clinical data of 19 children diagnosed with ALGS in Pediatric Gastroenterology Department, Shengjing Hospital of China Medical University since June 2013 to December 2022 was retrospectively analyzed, and the clinical manifestations of various systems of the body were followed up and evaluated, and then developed the personalised management strategies.Results:Among the 19 confirmed patients, 18 cases were confirmed by genetic testing.Eighteen cases(94.7%) had characteristic facial features.To follow-up node, 8 cases(42.1%) had cholestasis, with alanine aminotransferase(210.20±110.50)U/L, aspartate aminotransferase(187.86±96.70)U/L, and direct bilirubin(110.93±108.15)μmol/L.Eighteen cases(94.7%) had pruritus.Eighteen cases(94.7%) of the patients had a high risk of malnutrition, and the level of total bilirubin[(76.17±107.34)μmol/L] and total bile acid[(100.18±83.78)μmol/L] were significantly increased in the children with obvious growth retardation.Thirteen cases(68.42%) had diffuse liver injury.The clinical opinions on genetic counseling, application of new drugs, liver transplantation, cardiac medicine and surgery follow-up, spine and oral surgery orthodontics were given by multiple disciplines.Conclusion:ALGS children have a high risk of long-term malnutrition and are associated with the severity of liver injury, and pruritus and jaundice are the main clinical manifestations.The management of ALGS patients should be centered around liver disease doctors, combined with multiple disciplines, paying attention to changes in various related organs of ALGS patients, and improving their quality of life.

The effect of anti-programmed cell death 1 (anti-PD-1) immunotherapy is limited in patients with hepatocellular carcinoma (HCC). Yes-associated protein 1 (YAP1) expression increased in liver tumor cells in early HCC, and Akkermansia muciniphila abundance decreased in the colon. The response to anti-PD-1 treatment is associated with A. muciniphila abundance in many tumors. However, the interaction between A. muciniphila abundance and YAP1 expression remains unclear in HCC. Here, anti-PD-1 treatment decreased A. muciniphila abundance in the colon, but increased YAP1 expression in the tumor cells by mice with liver tumors in situ. Mechanistically, hepatocyte-specific Yap1 knockout (Yap1^LKO) maintained bile acid homeostasis in the liver, resulting in an increased abundance of A. muciniphila in the colon. Yap1 knockout enhanced anti-PD-1 efficacy. Therefore, YAP1 inhibition is a potential target for increasing A. muciniphila abundance to promote anti-PD-1 efficacy in liver tumors. Dihydroartemisinin (DHA), acting as YAP1 inhibitor, increased A. muciniphila abundance to sensitize anti-PD-1 therapy. A. muciniphila by gavage increased the number and activation of CD8⁺ T cells in liver tumor niches during DHA treatment or combination with anti-PD-1. Our findings suggested that the combination anti-PD-1 with DHA is an effective strategy for liver tumor treatment.