OBJECTIVE: To reveal the molecular basis of knee osteoarthritis (KOA) with Yang deficiency and blood stasis syndrome by analyzing the gene expression profiles in synovial fluid and blood of KOA patients with this syndrome. METHODS: A total of 80 KOA patients were recruited from October 2022 to June 2024, including 40 cases in the non-Yang deficiency and blood stasis group (27 males and 13 females), with an average age of (61.75±3.45) years old;and 40 cases in the Yang deficiency and blood stasis group (22 males and 18 females), with an average age of (62.00±2.76) years old. The levels of body mass index (BMI), high-density lipoprotein (HDL), low-density lipoprotein (LDL), fibrinogen, total cholesterol, and D-dimer were recorded and summarized. Blood and synovial fluid samples from patients were collected for gene expression profile microarray sequencing, and then PCR and immunohistochemistry were used for clinical verification on the patients' synovial fluid and cartilage samples. RESULTS: Logistic regression analysis showed that compared with KOA patients with non-Yang deficiency and blood stasis syndrome, those with Yang deficiency and blood stasis syndrome had increased BMI, LDL, fibrinogen, total cholesterol, and D-dimer, and decreased HDL, with a clear correlation between the two groups. There were 562 differential genes in the blood, among which 322 were up-regulated and 240 were down-regulated;755 differential genes were found in the synovial fluid, with 350 up-regulated and 405 down-regulated. KEGG signaling pathway analysis of synovial fluid revealed changes in lipid metabolism-related pathways, including cholesterol metabolism, fatty acid metabolism, and PPARG signaling pathway. Analysis of the involved differential genes identified 6 genes in synovial fluid that were closely related to lipid metabolism, namely LRP1, LPL, ACOT6, TM6SF2, DGKK, and PPARG. Subsequently, PCR and immunohistochemical verification were performed using synovial fluid and cartilage samples, and the results were consistent with those of microarray sequencing. CONCLUSION This study explores the clinical and genomic correlation between traditional Chinese medicine syndromes and knee osteoarthritis from the perspective of lipid metabolism, and proves that abnormal lipid metabolism is closely related to KOA with Yang deficiency and blood stasis syndrome from both clinical and basic aspects.
Humans ; Male ; Female ; Middle Aged ; Synovial Fluid/metabolism* ; Osteoarthritis, Knee/metabolism* ; Yang Deficiency/complications* ; Aged

Given that ovarian stimulation is vital for assisted reproductive technology (ART) and results in elevated serum estrogen levels, exploring the impact of elevated estrogen exposure on oocytes and embryos is necessary. We investigated the effects of various ovarian stimulation treatments on oocyte and embryo morphology and gene expression using a mouse model and estrogen-treated mouse embryonic stem cells (mESCs). Female C57BL/6J mice were subjected to two types of conventional ovarian stimulation and ovarian hyperstimulation; mice treated with only normal saline served as controls. Hyperstimulation resulted in high serum estrogen levels, enlarged ovaries, an increased number of aberrant oocytes, and decreased embryo formation. The messenger RNA (mRNA)‍-sequencing of oocytes revealed the dysregulated expression of lysine-specific demethylase 6b (Kdm6b), which may be a key factor indicating hyperstimulation-induced aberrant oocytes and embryos. In vitro, Kdm6b expression was downregulated in mESCs treated with high-dose estrogen; treatment with an estrogen receptor antagonist could reverse this downregulated expression level. Furthermore, treatment with high-dose estrogen resulted in the upregulated expression of histone H3 lysine 27 trimethylation (H3K27me3) and phosphorylated H2A histone family member X (γ-H2AX). Notably, knockdown of Kdm6b and high estrogen levels hindered the formation of embryoid bodies, with a concomitant increase in the expression of H3K27me3 and γ-H2AX. Collectively, our findings revealed that hyperstimulation-induced high-dose estrogen could impair the demethylation of H3K27me3 by reducing Kdm6b expression. Accordingly, Kdm6b could be a promising marker for clinically predicting ART outcomes in patients with ovarian hyperstimulation syndrome.
Female ; Mice ; Demethylation/drug effects* ; Embryonic Stem Cells ; Estrogens/administration & dosage* ; Gene Expression/drug effects* ; Histones/metabolism* ; Jumonji Domain-Containing Histone Demethylases/metabolism* ; Mice, Inbred C57BL ; Oocytes ; Ovary/drug effects* ; Reproductive Techniques, Assisted ; Animals

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 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.

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 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 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.

AIM To study the chemical constituents from the branches and leaves of Toona ciliata Roem.var.pubescens(Franch.)Hand-Mazz.and their antitumor activities.METHODS The compounds were isolated and purified by silica gel,RP-18 reverse phase silica gel and semi-preparative HPLC,the structures of compounds were identified by physicochemical properties and spectral data.The antitumor activities were determined by MTT method.RESULTS Fifteen compounds were isolated and identified as toonaolide D(1),toonaciliatin E(2),bourjotinolone A(3),(21R,23R)-epoxy-21α-ethoxy-24S,25-dihydroxyapotirucalla-7-en-3-one(4),(Z)-toonasterone C(5),(E)-toonasterone(6),3-epi-dyscusin C(7),(Z)-aglawone(8),(E)-volkendousin(9),8(14),15-isopimaradiene-2α,3α,19-triol(10),(-)-loliolide(11),cyclohexenone(12),pubinernoid A(13),quercetin-3-O-(4″-methoxy)-α-L-rahmnopyranosyl(14),5-hydroxymethyl-2-furancarboxaldehyde(15).The IC50 values of compounds 3 and 4 on K562 cells were 54.2 and 47.3 μmol/L,respectively,and the IC50 values on HEL cells were 47.3 and 61.1 μmol/L,respectively.CONCLUTION Compounds 4,7,10 and 11 are isolated from Toona genus for the first time,and compounds 2,15 are first isolated from this plant.Compounds 3 and 4 show weak antitumor activities.

Objective:To compare the safety and efficacy of two-person and three-person operation of endoscopic retrograde cholangiopancreatography (ERCP) after Roux-en-Y anastomosis.Methods:A total of 31 cases of bile duct stones where endoscopic access was successful and papilla was detected after Roux-en-Y anastomosis in Hangzhou First People's Hospital from January 2021 to September 2022 were recruited. Based on the retrospective study, 17 patients with the two-person operation (an operator and an assistant completed selective intubation) were included in group A from January to December 2021, and 14 patients with the three-person operation (an operator and two assistants completed the selective intubation) from January to September 2022 were included in group B. The success rate, the time of intubation, and complication incidence of the two groups were compared.Results:The intubation success rate of all enrolled patients was 90.32% (28/31). There was no significant difference in the intubation success rate between group A and group B [82.35% (14/17) VS 100.00% (14/14), P=0.232]. The intubation time in group B was significantly less than that of group A (42.89±6.57 min VS 61.02±9.23 min) with significant difference ( t=5.77, P<0.001). The incidence of postoperative complication of ERCP in all enrolled patients was 16.13% (5/31). There was no significant difference in the postoperative complication incidence between group A and group B [17.65% (3/17) VS 14.29% (2/14), P=1.000]. Conclusion:For patients undergoing ERCP after Roux-en-Y anastomosis, the two methods were not statistically different in terms of the intubation success rate and the incidence of procedure-related complications. But three-person operation shows clear advantage in terms of operation time.

Objective:To explore the impact of the hospital health literacy environment on patients′ postoperative pain self-management behaviors, aiming to provide insights for hospitals to implement the Comprehensive Pain Management Pilot Work Program in hospitals and to promote self-health management among patients with other diseases or symptoms. Methods:From November to December 2023, a convenience sampling method was used to select postoperative patients from three grade A tertiary general hospitals in Zhejiang Province for an on-site questionnaire survey. The Chinese version of brief health literacy screen (BHLS), short-form health literacy environment scale (SF-HLES) and postoperative pain self-management behavior questionnaire (PPSMB) were used as survey tools to investigate the health literacy level of patients, the health literacy environment of the hospital, and the postoperative pain management behaviors of patients. Two-way ANOVA was used to compare the impact of different dimensions of the hospital health literacy environment on postoperative pain management behaviors among patients with different levels of health literacy. Multiple linear regression analysis was used to explore the relationship between the hospital health literacy environment, individual health literacy, and patients′ postoperative pain self-management behaviors, and to discuss the impact of individual health literacy on patients′ postoperative pain self-management behaviors under different hospital health literacy environments.Results:341 valid questionnaires were collected. The average score of the hospitals′ SF-HLES was (73.62±19.54) points. The average score of the patients′ BHLS was (9.65±2.88) points. The average score of the patients′ PPSMB was (25.99±6.35) points. Two-way ANOVA results showed that the interaction between individual health literacy and the clinical dimension ( F=5.463, P=0.020) and structural dimension ( F=6.470, P=0.011) of the hospital health literacy environment had a statistically significant impact on patients′ postoperative pain self-management behaviors, while the interaction with the interpersonal dimension ( F=0, P=0.984) had no statistically significant impact on pain self-management behaviors. Simple effect analysis indicated that only in the high health literacy environment of the clinical and structural dimensions did the difference in pain self-management behaviors between patients with good health literacy and those with limited health literacy had statistical significance ( P<0.001). Multiple linear regression analysis results showed that for each 1-point increase in the patients′ BHLS score, their PPSMB score increased by 3.74 points ( β1=0.832, P<0.001); for each 1-point increase in the hospital′s SF-HLES score, the patients′ PPSMB score could increase by 0.198 points ( β2=0.610, P<0.001). In a low health literacy environment, individual health literacy did not affect pain self-management behaviors ( P>0.05); however, in a high health literacy environment, for each 1-point increase in the patients′ BHLS score, their PPSMB score correspondingly increased by 4.037 points ( β4=0.317, P<0.001). Conclusions:The positive impact of individual health literacy on pain self-management is contingent upon a high-quality hospital health literacy environment. This suggests that optimizing the hospital health literacy environment is a necessary precondition for implementing the relevant content of the Comprehensive Pain Management Pilot Work Program and can provide a reference for promote self-health management among patients with pain and other diseases or symptoms.