The aim of this study was to investigate the virulence determinants and genetic diversity of foodborne Yersinia enterocolitica from Wenzhou.A total of 71 strains of Yersinia enterocolitica were isolated from food and foodborne diarrhea ca-ses in Wenzhou,and their biotypes,serotypes,and drug resistance were analyzed.On the basis of whole genome sequencing,we assessed virulence gene profiles,and performed multilocus sequence typing(MLST)and core gene multilocus sequence typ-ing(cgMLST).A total of 94.4％(67/71)of isolates belonged to biotype 1A,and the highest proportion had serotype lA/O∶5(29.6％,21/71).The sensitivity rates of the isolates to 14 antibiotics exceeded 95.8％.A total of 16 categories and 126 viru-lence genes were identified,with two strains carrying the pYV plasmid and chromosome-related virulence genes.ST3(31.6％,12/38)was the most widespread MLST type,and cgMLST analysis revealed no dense clusters of genotypes except for strains sharing the same ST.In conclusion,pathogenic strains were identified from foodborne Yersinia enterocolitica in Wenzhou and were found to exhibit high genetic polymorphism.Enhanced regulatory supervision is essential to prevent the outbreak of food-borne diseases caused by Yersinia enterocolitica.

Objective To evaluate the effects of puerarin on bone density in rats and mice through a meta-analysis. Methods The databases, including CNKI, SinoMed, Wanfang data, VIP, PubMed, EMBase, Web of Science, the Cochrane Library, and Scopus from their inception to November 6, 2023, were searched for literature on the effects of puerarin treatment on bone density in rats and mice. Inclusion criteria for the literature were randomized controlled trials with a placebo or blank control group; the subject animals were rats or mice; the intervention was puerarin; and the results included bone density measurements. Exclusion criteria included combination therapy with puerarin; lack of original research data; unpublished studies; and using mandible as the measurement site for bone density. Risks of bias were assessed using SYRCLE's RoB tool. Data analysis was conducted with Stata 16.0 and Rev Man 5.3 software. Results After applying the inclusion and exclusion criteria, a total of 429 records were identified and 42 articles covering 41 studies were ultimately included. 925 animals were involved and the data analysis results indicated that puerarin improved bone density in rats and mice compared to the control group: femur [37 studies, n=824, standardized mean difference （SMD)=2.12, 95% confidence interval (CI)=1.69-2.54, P < 0.000 1], lumbar spine (13 studies, n=271, SMD=2.25, 95% CI=1.49-3.01, P < 0.000 1), tibia (4 studies, n=95, SMD=0.94, 95% CI=0.05-1.83, P=0.04), and the whole body (4 studies, n=94, SMD=1.89, 95% CI=0.50-3.29, P=0.008), with all inter-group differences in bone density being statistically significant. Conclusion Puerarin can improve bone density in rats and mice. This study provides a valuable reference for clinical studies on the prevention and treatment of osteoporosis with puerarin.

BACKGROUND:Resveratrol is a natural antioxidant extracted from plants.Its mechanism of improving exercise-induced fatigue mainly focuses on the protective effect against oxidative stress and inflammation.In this study,the protective mechanism of resveratrol on exercise-induced fatigue was mainly discussed from the perspective of gluconeogenesis. OBJECTIVE:To investigate the effect of resveratrol on gluconeogenesis in exercise-induced fatigue rats. METHODS:After 1 week of adaptive training,male Sprague-Dawley rats were randomly divided into 4 groups with 12 rats in each group:blank control group,resveratrol group,exercise group,resveratrol + exercise group.Weight-bearing swimming training was used to simulate long-term medium-high intensity exercise.After swimming with a weight of 5%for 1 hour every day,50 mg/kg resveratrol solution or the same volume of dimethyl sulfoxide solvent were given orally,6 days a week,for a total of 6 weeks.Samples were collected 24 hours after the last exercise,and the levels of urea nitrogen,creatine kinase,blood glucose,liver glycogen and lactic acid and pyruvate in liver tissue were detected by the kit.The activity of phosphoenolpyruvate carboxykinase was detected by microassay,and the activity of glucose-6-phosphatase was detected by enzyme-linked immunosorbent assay.Real-time fluorescence quantitative PCR was used to detect the gene expression of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α. RESULTS AND CONCLUSION:In the exercise group,plasma urea nitrogen and creatine kinase levels of rats were significantly increased(both P<0.05),liver lactate and pyruvate levels and lactate/pyruvate ratio were significantly increased(all P<0.01),and blood glucose and liver glycogen contents were significantly decreased(both P<0.01).Resveratrol supplementation could effectively improve the above conditions.Exercise significantly decreased the activities of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase(P<0.01,P<0.05),and resveratrol supplementation significantly increased the activity of phosphoenolpyruvate carboxykinase in liver tissue(P<0.01).The mRNA expression levels of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α in liver tissue of the exercise group were significantly decreased(all P<0.01),while resveratrol supplementation could significantly increase the gene expression levels of this pathway.To conclude,resveratrol can relieve exercise-induced fatigue caused by long-term medium-high intensity exercise,and its mechanism may be related to up-regulating the gluconeogenesis regulatory pathway,improving rate-limiting enzyme activity,promoting liver gluconeogenesis,and increasing blood glucose and liver glycogen levels.

Objective To study the bioequivalence and safety of two olmesartan medoxomil and hydrochlorothiazide tablets in Chinese healthy subjects.Methods A total of 24 healthy subjects underwent fasting and postprandial tests in a single-center,randomized,open-label,single-dose,two-formulation,two-sequence,two-period,self-cross-over controlled design.The subjects were administered a single oral dose of the test formulation and reference formulation(each containingolmesartan medoxomil 20 mg and hydrochlorothiazide 12.5 mg)in a random cross-over fashion.The plasma concentrations of olmesartan and hydrochlorothiazide were determined by LC-MS/MS.The non-compartmental model analysis of olmesartan and hydrochlorothiazide was conducted using WinNonlin 7.0 software to calculate pharmacokinetic parameters and assess bioequivalence.Results In the fasting test,the pharmacokinetic parameters of olmesartan of test and reference were as follows:Cmax were(798.35±206.78)and(664.52±168.25)ng·mL-1,AUC0-t were(4 430.71±1 294.87)and(3 976.67±1 083.54)h·ng·mL-1,AUC0-∞ were(4 551.67±1 303.06)and(4 090.37±1 103.97)h·ng·mL-1.The pharmacokinetic parameters of hydrochlorothiazide of test and reference were as follows:Cmax were(92.39±35.96)and(96.15±38.76)ng·mL-1,AUC0_t were(548.69±217.11)and(564.41±208.68)h·ng·mL-1,AUC0-∞ were(603.04±228.59)and(619.26±223.27)h·ng·mL-1.In the fed test,the pharmacokinetic parameters of olmesartan of T and R were as follows:Cmax were(583.15±149.48)and(550.57±104.76)ng·mL-1,AUC0-t were(3 585.18±952.72)and(3 292.19±904.58)h·ng·mL-1,AUC0-∞ were(3 696.05±996.55)and(3 396.30±923.41)h·ng·mL-1.The pharmacokinetic parameters of hydrochlorothiazide of test and reference were as follows:Cmax were(70.30±17.88)and(74.70±21.65)ng·mL-1,AUC0-t were(476.60±119.39)and(492.91±144.81)h·ng·mL-1,AUC0-∞ were(523.37±132.67)and(535.81±151.92)h·ng·mL-1.In fasting and fed condition,the 90％confidence interval(90％CI)of Cmax,AUC0-t and AUC0-∞ of olmesartan and hydrochlorothiazide were in 80.00％-125.00％.Conclusion The two olmesartan medoxomil and hydrochlorothiazide tablets were bioequivalent under fasting and fed conditions,and good security.

Objective:To assess the accuracy of two-dimensional (2D) photographs in measuring esthetic parameters of the maxillary anterior teeth by comparing them with measurements obtained from three-dimensional (3D) dental models.Methods:A total of one hundred volunteers (49 males, 51 females, aged 18-23 years) were recruited from School and Hospital of Stomatology, Wuhan University from January to February 2024. 3D digital models of their dentitions were obtained using an intraoral scanner, and standardized frontal 2D intraoral photographs were captured with a digital camera. The lengths, widths and width/length ratio of the bilateral incisors, lateral incisors and canines were measured on both the 3D digital models and the 2D intraoral photographs. The width ratios of adjacent maxillary anterior were also calculated on the 2D intraoral photographs and the frontal view of 3D digital models.Results:The widths of lateral incisors [(5.85±0.60) mm] and canines [(4.73±0.71) mm] and the lengths of canines [(8.72±0.96) mm] in the 2D intraoral photographs were significantly lower than those in 3D digital models [(6.65±0.59), (7.76±0.60), (8.90±0.86) mm] ( t=-18.24, P<0.001; t=-54.43, P<0.001; t=-4.40, P<0.001), while there were no significant differences in the lengths and widths of the other teeth ( P>0.05). The width/length ratios measured from the 2D intraoral photographs for the lateral incisors and canines (0.74±0.08, 0.55±0.08) were significantly lower than those measured in the 3D digital models (0.84±0.09, 0.88±0.09) ( t=-19.68, P<0.001; t=-50.21, P<0.001), and the width/length ratio of the central incisors showed no significant difference between the two groups ( P>0.05). The width ratios of canines/lateral incisors and lateral incisors/central incisors measured on the 2D intraoral photographs (0.72±0.06, 0.85±0.11) were significantly smaller than those measured in the frontal view of 3D digital models (0.75±0.06, 0.89±0.11) ( t=-9.31, P<0.001; t=-6.58, P<0.001). Conclusions:There is a difference between 2D and 3D measurement results of teeth in the esthetic area and the magnitude of the difference varies with their position in the dental arch. When analyzing the measurement of the anterior teeth, it is necessary to choose the appropriate method according to the target tooth position.

Objective:To investigate the role and mechanism of human umbilical cord mesenchymal stem cell exosomes (hUCMSC-ex) in wounds with escharectomy and skin grafting in scalded rats.Methods:The study was an experimental study. Twelve male Sprague-Dawley (SD) rats aged 6-8 weeks were divided into combined treatment group, fixed+allogeneic skin group, autologous skin+allogeneic skin group, and allogeneic skin group by random number table method (the same grouping method hereinafter), with 3 rats in each group. The four groups of rats were inflicted with scalded wounds on the back and performed with escharectomy, and then the wounds of rats in combined treatment group were fixed with a metal ring (the same fixing method hereinafter) and transplanted with autologous skin grafts and allogeneic skin grafts, and the other three groups of rats were fixed and/or transplanted with skin grafts corresponding to the group name. At 14, 21, and 28 d after surgery, the wound healing area in the four groups of rats was measured. Another 15 male SD rats aged 6-8 weeks were divided into normal group with no treatment, high exosome group, low exosome group, supernatant group, and phosphate buffer solution (PBS) group, with 3 rats in each group. The last 4 groups of rats were treated as that in the above-mentioned combined treatment group, and then were injected around the wounds with 200 μL of PBS containing 100 μg of hUCMSC-ex, 200 μL of PBS containing 50 μg of hUCMSC-ex, 200 μL of supernatant with no hUCMSC-ex, and 200 μL of PBS at 0 (immediately), 7, 14, and 21 d after surgery, respectively. At 14, 21, and 28 d after surgery, the wound healing area in the four groups of rats was measured. The wound neo-epithelial tissue of rats in high exosome group and PBS group at 28 d after surgery and the normal skin tissue of rats in normal group at the same time point were taken, and the differentially expressed proteins were screened by label-free quantitative proteomics method; the two up-regulated and differentially expressed proteins, the immunoglobulin G1 heavy chain constant region (IGHG1) and cystatin A (CSTA) with the largest and second largest fold changes in comparison between high exosome group and PBS group were selected, and their protein expressions were detected by Western blotting. The number of samples in all experiments was 3.Results:At 14, 21, and 28 d after surgery, the wound healing area in combined treatment group, autologous skin+allogeneic skin group, and allogeneic skin group of rats was significantly larger than that in fixed+allogeneic skin group ( P<0.05), the wound healing area in autologous skin+allogeneic skin group of rats at 21 d after surgery and that in allogeneic skin group of rats at 14 and 21 d after surgery was significantly larger than that in combined treatment group ( P<0.05), and the wound healing area in allogeneic skin group of rats at 14 d after surgery was significantly larger than that in autologous skin+allogeneic skin group ( P<0.05). The wound healing area of rats in high exosome group and low exosome group at 14, 21, and 28 d after surgery and in supernatant group at 14 and 28 d after surgery was significantly larger than that in PBS group ( P<0.05); the wound healing area in high exosome group of rats at 14 and 21 d after surgery was significantly larger than that in supernatant group ( P<0.05), and the wound healing area at 14 d after surgery was significantly larger than that in low exosome group ( P<0.05); the wound healing area in low exosome group of rats at 14 d after surgery was significantly larger than that in supernatant group ( P<0.05). Compared with that in PBS group, 332 proteins were differentially expressed in the neo-epithelial tissue of the wounds in high exosome group of rats at 28 d after surgery ( P<0.05), among which the protein expressions of IGHG1 and CSTA were significantly up-regulated (with fold change of 12.60 and 2.27, respectively, P<0.05). Compared with those of normal skin tissue in normal group, 1 400 and 1 057 proteins were differentially expressed in the neo-epithelial tissue of the wounds in high exosome group and PBS group of rats at 28 d after surgery, respectively. The protein expressions of IGHG1 and CSTA in the wound neo-epithelial tissue in high exosome group of rats at 28 d after surgery were significantly larger than those in normal skin tissue of rats in normal group ( P<0.05) and those in PBS group ( P<0.05). Conclusions:hUCMSC-ex may accelerate the repair process of wounds with escharectomy and skin grafting and improve the quality of wound healing in scalded rats by regulating the protein expressions of IGHG1 and CSTA.

Objective To evaluate the application value of the Global Leadership Initiative on Malnutri-tion(GLIM)criteria in the patients with gastric cancer radical surgery,and to explore the relationship be-tween the malnutrition defined by GLIM criteria and the prognosis of the patients with gastric cancer radical surgery.Methods A total of 150 gastric cancer patients receiving radical surgical resection in Zhejiang Provin-cial Tongde Hospital from January 2014 to December 2017 were selected as the study subjects.All the study subjects were diagnosed as malnutrition and graded by the severity of malnutrition.The effect of malnutrition defined by GLIM criteria on the prognosis of the patients with gastric cancer radical surgery was evaluated by the Kaplan Meier survival curve and Cox regression analysis.The receiver operating characteristic(ROC)curve was used to evaluate the predictive value of GLIM criteria on the postoperative survival rate of the pa-tients with gastric cancer radical surgery,and the corresponding area under the curve(AUC)was calculated,and the predictive value of inflammatory indicator C-reactive protein(CRP)addition on GLIM criteria in pre-dicting the prognosis of the patients with gastric cancer radical surgery was evaluated.Results Malnutrition diagnosed by GLIM criteria was an independent risk factor affecting the postoperative survival rate of the pa-tients with gastric cancer radical surgery.The results of survival trend analysis of various indicators in GLIM criteria showed that involuntary weight reduction(moderate malnutrition HR=14.13,95％CI:1.70-117.39,severe malnutrition HR=12.50,95％CI:1.40-111.89)and CRP＞10 mg/L(HR=9.70,95％CI:2.31-40.67)were the most important factors affecting the survival rate of the patients with gastric cancer radical surgery.In addition,the addition of inflammatory marker CRP could increase the sensitivity and speci-ficity of the GLIM model in predicting the postoperative survival rate of the patients with gastric cancer radical sur-gery.Conclusion The GLIM criteria can not only reflect the nutritional status of the patients with gastric cancer radi-cal surgery,but also serve as an effective predictive tool for predicting the survival rate after radical surgery.The addition of inflammatory marker CRP increases the sensitivity and specificity of GLIM criteria in predicting postoperative survival rate.

Objective To construct a prognostic model for liver hepatocellular carcinoma(LIHC)based on immune-related genes.Methods LIHC and normal tissue samples were downloaded from the UCSC Xena database and The Cancer Genome Atlas(TCGA)database.Differential analysis was performed on the gene data of LIHC samples and adjacent/normal samples.Enrichment analysis was conducted on differentially expressed genes.Kaplan-Meier survival analysis was performed on liver cancer samples from the TCGA cohort to obtain survival-and immune-related differentially expressed genes.LASSO Cox and multivariate Cox regression analysis were used to identify hub genes and construct a gene risk prognostic model.Data from a high-throughput gene expression(GEO)database was obtained for external validation.The sensitivity of hub genes to common anticancer drugs was investigated using the CellMiner database.Results Enrichment analysis result indicated that differentially expressed genes were mainly associated with metabolic pathways.Through differential analysis and Kaplan-Meier survival analysis,25 survival-and immune-related differentially expressed genes were obtained.Based on the result of LASSO Cox and multivariate Cox regression analysis,five hub genes(FYN,CSF3R,HLA-G,FOS,BIRC5)were identified and a nomogram was constructed.The concordance index(C-index)value for the training cohort and validation cohort were 0.739 and 0.625,respectively.Based on the sensitivity of hub genes to anticancer drugs,12 types of anticancer drugs were selected for subsequent experiments.Conclusion This model can effectively predict the prognosis of LIHC patients and provide a new insights for immune therapy in LIHC.

Objective:To compare the application effect of domestic and imported intravenous radiofrequency closure system in the treatment of primary varicose veins of lower extremities.Methods:This single-center prospective, non-inferiority randomized controlled trial was performed in the Department of Vascular Surgery, the Fourth Affiliated Hospital, Zhejiang University School of Medicine from January 2021 to January 2022. Patients with primary varicose veins of lower extremities who met the ataxation criteria were randomly assigned to the experimental group(domestic novel venous radiofrequency closure system) or the control group(imported venous radiofrequency closure system) in a ratio of 1∶1. The two groups of subjects were compared in terms of target vein closure rate, technical success rate, system operation performance, incidence of adverse events and incidence of serious adverse events(SAE) within 6 months after surgery. Quantitative data were compared by Mann-Whitney U test, and categorical data were compared by χ2 test and non-inferiority test. Results:A total of 80 subjects were included in the trial (41 in the experimental group and 39 in the control group), including 27 males and 53 females, aged ( M(IQR)) 55(23) years (range:40 to 78 years). There were 48 cases of left lower limb and 32 cases of right lower limb. The technical success rate and system control performance between the groups were 100%.The incidence of adverse events (58.5% (24/41) vs. 61.5% (24/39), χ2=0.075, P=0.784), and the incidence of SAE (7.3% (3/41) vs. 5.1% (2/39), χ2=0.163, P=0.686) within 6 months after surgery in experimental group and control group had no statistical significance. There was one device-related adverse event in each of the two groups. In the experimental group, one patient developed endovenous heat-induced thrombosis after surgery and recovered after taking rivaroxaban tablets. One patient in the control group had pain in the upper right thigh for more than 1 day after operation, which was cured after using analgesic cream. No device-related SAE occurred. The venous closure rate of the experimental group was 100% (38/38) at 6 months after surgery, and that of the control group was 97.4% (37/38). The difference between the two groups was 2.63% (95% CI:-3.19 to 8.45, Z=4.865, P<0.01), and the 95% CI lower limit of the difference in target venous closure rate between two groups was greater than the non-inferiority threshold of -10.00%. Conclusion:The early application effect of the new domestic intravenous radiofrequency closure system in patients with primary varicose veins of lower extremities is in line with expectations, it is not inferior to the imported system.

Objective:To compare the application effect of domestic and imported intravenous radiofrequency closure system in the treatment of primary varicose veins of lower extremities.Methods:This single-center prospective, non-inferiority randomized controlled trial was performed in the Department of Vascular Surgery, the Fourth Affiliated Hospital, Zhejiang University School of Medicine from January 2021 to January 2022. Patients with primary varicose veins of lower extremities who met the ataxation criteria were randomly assigned to the experimental group(domestic novel venous radiofrequency closure system) or the control group(imported venous radiofrequency closure system) in a ratio of 1∶1. The two groups of subjects were compared in terms of target vein closure rate, technical success rate, system operation performance, incidence of adverse events and incidence of serious adverse events(SAE) within 6 months after surgery. Quantitative data were compared by Mann-Whitney U test, and categorical data were compared by χ2 test and non-inferiority test. Results:A total of 80 subjects were included in the trial (41 in the experimental group and 39 in the control group), including 27 males and 53 females, aged ( M(IQR)) 55(23) years (range:40 to 78 years). There were 48 cases of left lower limb and 32 cases of right lower limb. The technical success rate and system control performance between the groups were 100%.The incidence of adverse events (58.5% (24/41) vs. 61.5% (24/39), χ2=0.075, P=0.784), and the incidence of SAE (7.3% (3/41) vs. 5.1% (2/39), χ2=0.163, P=0.686) within 6 months after surgery in experimental group and control group had no statistical significance. There was one device-related adverse event in each of the two groups. In the experimental group, one patient developed endovenous heat-induced thrombosis after surgery and recovered after taking rivaroxaban tablets. One patient in the control group had pain in the upper right thigh for more than 1 day after operation, which was cured after using analgesic cream. No device-related SAE occurred. The venous closure rate of the experimental group was 100% (38/38) at 6 months after surgery, and that of the control group was 97.4% (37/38). The difference between the two groups was 2.63% (95% CI:-3.19 to 8.45, Z=4.865, P<0.01), and the 95% CI lower limit of the difference in target venous closure rate between two groups was greater than the non-inferiority threshold of -10.00%. Conclusion:The early application effect of the new domestic intravenous radiofrequency closure system in patients with primary varicose veins of lower extremities is in line with expectations, it is not inferior to the imported system.