Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder affecting a substantial proportion of women of reproductive age. It is frequently associated with ovulatory dysfunction, infertility, and an increased risk of chronic metabolic diseases. A hallmark pathological feature of PCOS is the arrest of follicular development, closely linked to impaired intercellular communication between the oocyte and surrounding granulosa cells. Transzonal projections (TZPs) are specialized cytoplasmic extensions derived from granulosa cells that penetrate the zona pellucida to establish direct contact with the oocyte. These structures serve as essential conduits for the transfer of metabolites, signaling molecules (e.g., cAMP, cGMP), and regulatory factors (e.g., microRNAs, growth differentiation factors), thereby maintaining meiotic arrest, facilitating metabolic cooperation, and supporting gene expression regulation in the oocyte. The proper formation and maintenance of TZPs depend on the cytoskeletal integrity of granulosa cells and the regulated expression of key connexins, particularly CX37 and CX43. Recent studies have revealed that in PCOS, TZPs exhibit significant structural and functional abnormalities. Contributing factors—such as hyperandrogenism, insulin resistance, oxidative stress, chronic inflammation, and dysregulation of critical signaling pathways (including PI3K/Akt, Wnt/β‑catenin, and MAPK/ERK)—collectively impair TZP integrity and reduce their formation. This disruption in granulosa-oocyte communication compromises oocyte quality and contributes to follicular arrest and anovulation. This review provides a comprehensive overview of TZP biology, including their formation mechanisms, molecular composition, and stage-specific dynamics during folliculogenesis. We highlight the pathological alterations in TZPs observed in PCOS and elucidate how endocrine and metabolic disturbances—particularly androgen excess and hyperinsulinemia—downregulate CX43 expression and impair gap junction function, thereby exacerbating ovarian microenvironmental dysfunction. Furthermore, we explore emerging therapeutic strategies aimed at preserving or restoring TZP integrity. Anti-androgen therapies (e.g., spironolactone, flutamide), insulin sensitizers (e.g., metformin), and GLP-1 receptor agonists (e.g., liraglutide) have shown potential in modulating connexin expression and enhancing granulosa-oocyte communication. In addition, agents such as melatonin, AMPK activators, and GDF9/BMP15 analogs may promote TZP formation and improve oocyte competence. Advanced technologies, including ovarian organoid models and CRISPR-based gene editing, offer promising platforms for studying TZP regulation and developing targeted interventions. In summary, TZPs are indispensable for maintaining follicular homeostasis, and their disruption plays a pivotal role in the pathogenesis of PCOS-related folliculogenesis failure. Targeting TZP integrity represents a promising therapeutic avenue in PCOS management and warrants further mechanistic and translational investigation.

OBJECTIVE To investigate the effect of leech freeze-dried powder combined with scorpion freeze-dried powder on carotid artery thrombosis in rats. METHODS FeCl3 was used to induce carotid artery thrombosis, and the experiment was divided into 6 groups: sham-operated group(sham group), model group, leech freeze-dried powder group(leech group), whole scorpion freeze-dried powder group(scorpion group), leech+scorpion half-dose group(half-dose group), leech+scorpion full-dose group(full-dose group), 6 rats in each group. The inhibition rate of bilateral carotid artery thrombosis was calculated after the final administration. The activated partial thromboplastin time(APTT), plasma prothrombintime(PT), plasma fibrinogen(FIB), and prothrombin time(TT) were measured by platelet aggregation and coagulation factor analyzer. The levels of plasma tissue-type plasminogen activator(t-PA) and plasminogen activatorinhibitor-I(PAI-I) were measured by ELSIA, and the thrombosis of each group was observed by HE staining. RESULTS Compared with model group, the wet weight and dry weight of thrombus were reduced in the leech, scorpion, half-dose and full-dose groups, and the full-dose group had the best effect in inhibiting thrombus formation. Compared with sham group, APTT, PT, TT was shortened, FIB content increased and t-PA level decreased in model group; compared with model group, APTT, PT, TT was prolonged, FIB content decreased and t-PA level increased in each dose group. The PAI-Ⅰ was not significantly different in each group. A large number of thrombus were seen in the blood vessels of model group. Compared with model group, there were significantly fewer thrombus in the leech, scorpion and half-dose groups, and almost no significant thrombus in the full-dose group. CONCLUSION Leech freeze-dried powder combined with scorpion freeze-dried powder inhibited carotid artery thrombosis in rats by affecting coagulation function and fibrinolytic system.

To investigate the differences in methylation levels of cuproptosis related genes in cervical cancer and their effects on clinical prognosis.Methods:The methylation data of 310 cervical tissue specimens were acquired from public databases. The UALCAN database was used to analyze the methylation level differences of 12 cuproptosis-related genes and study their level in different stages or grades of cervical cancer. Genes with statistically significant differences were selected for prognosis analysis using the EWAS datahub. Finally, gene-enrichment analysis, pathway analysis, immune infiltration analysis, the mutation rate and tumor mutation burden (TMB) of the genes in cervical cancer were analyzed using the cBioportal database. Two independent samples rank-sum test was used for differences in methylation levels and immune cell infiltration; comparative analyses of overall survival were performed using KM survival curves and Log-rank two-sided tests. TMB analyses were performed using the Wilcoxon Test for statistical analyses; Pearson correlation analysis was used for assessment in GSEA and pathway analyses.Results:The methylationβvalue of Cyclin Dependent Kinase Inhibitor 2A (CDKN2A gene) in the cervical cancer tissues of patients was 0.075 which was significantly higher than the methylationβvalue of 0.049 in normal human tissues ( P=0.008). Dihydrolipoamide S-Acetyltransferase (DLAT gene) methylation with a β value of 0.102 was significantly higher than normal human tissue methylation with a β value of 0.08 ( P=0.002), and the methylation level β value of Lipoyltransferase 1 (LIPT1 gene) in cervical cancer tissues was 0.06,which was significantly lower than normal human tissue methylation value of 0.092 ( P=0.009). Patients with CDKN2A gene methylation levels≥0.199 had an overall survival of 14.75 years, which was lower than that of patients with methylation levels<0.199 (17.56 years) ( P=0.034).The results of gene enrichment analysis indicated that it mainly involves biological processes such as the response to type I interferon and DNA replication. The expression of CDKN2A gene is positively correlated with the number of neutrophils and dendritic cells in the tumor microenvironment( P<0.05), and negatively correlated with the number ofmacrophages( P<0.05). TMB was higher in the group of variants of the CDKN2A gene than in the group of non-variants ( P=0.019). Conclusion:CDKN2A methylation is a potential biomarker for predicting the prognosis of cervical cancer.

Objective To investigate the mid-term effect and complications of arthroscopic popliteal tendon suture in the treatment of lateral meniscus injury.Methods From January 2016 to December 2020,the data of 57 patients with lateral meniscus popliteal tendon injury treated by arthroscopic popliteal tendon suture fixation were retrospectively analyzed,includ-ing 35 males and 22 females,aged from 18 to 47 years old with an average of(32.9±7.9)years old.Knee function was evaluat-ed using the International Knee Documentation Committee(IKDC)and Lysholm scores both before the operation and at the fi-nal follow-up.Meniscus healing was evaluated according to the postoperative Barrett standard.Wound healing complications,such as vascular injury,nerve injury,and lower extremity venous thrombosis,were recorded.Results All 57 patients were fol-lowed up for 12 to 58 months with an average of(38.1±14.9)months.The incisions of the patients after the operation were all Grade A healing without infection,popliteal tendon injury,blood vessel injury,nerve injury and lower extremity venous throm-bosis.The IKDC score increased from(49.7±3.6)points preoperatively to(88.5±4.4)points in the final follow-up(P＜0.05).The Lysholm score increased from(48.8±4.9)points preoperatively to(91.9±3.9)points at the final follow-up(P＜0.05).At 3,6 months and 1 year after operation,according to Barrett's criteria,54 cases were clinically healed,the healing rate was 94.7％(54/57).Conclusion This study preliminarily confirmed that arthroscopic suture technique can result in clinical sta-bility through suture and fixation of the meniscus in the injured lateral popliteal tendon area.No adverse effects on knee joint function were found in the mid-term follow-up after the operation.

Objective Based on the theory of Information-Knowledge-Attitude-Practice(IKAP),the reliability and validity of the discharge preparation scale for patients with diabetic foot(DF)was developed and tested,which is in accordance with the national conditions and disease characteristics of our country.Methods Based on the IKAP theoretical framework,articles were identified through literature analysis,qualitative interviews and Delphi method.After pre-investigation,a total of 483 patients with DF were selected from 3 general hospitals in Jiangxi province from June 2023 to February 2024.The items were screened by item analysis,and the reliability and validity were tested.Results The discharge preparation scale for patients with DF included 4 dimensions and 29 items.The Cronbach's α coefficient was 0.934;the scores of each dimension Cronbach's α coefficient ranged from 0.889 to 0.947;the overall split-half reliability was 0.966,and the split-half reliability of each dimension ranged from 0.901 to 0.952.A total of 4 common factors were extracted by exploratory factor analysis,and the cumulative contribution rate of variance was 68.074％.The x2/df value of confirmatory factor analysis was 1.906.The fitting fit of the model is suitable.The content validity was 0.973 at scale level and 0.875～1.000 at item level.Conclusion The reliability and validity of the discharge preparation scale for patients with DF are good,and it can be used as a tool to evaluate the discharge preparation for patients with DF.

Objective To investigate the effects of Anmian Dan on neurotransmitters in the brain of model rats,which were sleep deprived by multi-platform water environment.Methods Fifty SD rats were randomly and evenly divided into 5 groups with 10 rats in each group,which were blank control group(Control group),Model group(Model group),Estazolam group(Estazolam group),low dose group(AMD-L group)and high dose group(AMD-H group).The rats were subjected to sleep deprivation in a multi-platform water environment for 20 hours per day for 21 days.The movement distance and movement time of rats at different time points were recorded by autonomous activity analyzer to evaluate the changes of autonomous activity.The contents of glutamic acid(Glu)and gamma-aminobutyric acid(GABA)were detected by ELISA,and the mRNA expression levels of NDRG2,GLT-1,GAD65 and GAD67 were detected by Real-time PCR.Western blot was used to detect the expressions of NDRG2,p-PI3K,p-Akt,GLT-1,GAD65 and GAD67.Results The Model group was more active than the Control group,and the concentration of GABA in the cortex of the Model group was decreased and the concentration of Glu was increased.The mrna and protein expression levels of NDRG2 in Model group were higher than those in Control group(P<0.01),but the mrna and protein expression levels of GLT-1,GAD65 and GAD67 in model group were lower than those in Control group(P<0.01).The protein expression levels of P-PI3K and P-AKT in the cortex of model group were significantly decreased(P<0.01).Compared with Model group,Anmeidan could reduce the autonomic activity of sleep deprived rats,increase the concentration of GABA,decrease the concentration of Glu in cortex(P<0.05),and increase the mrna relative expression levels and protein expression levels of GLT-1,GAD65 and GAD67(P<0.05).The expression levels of P-PI3K and P-Akt were increased(P<0.01),and mrna and protein expression levels of NDRG2 were decreased(P<0.01).Conclusion Anmian Dan may regulate the activity of astrocytes and affect the levels of neurotransmitters GABA and GLU in the brain through the PI3K/AKT signaling pathway,thus playing a role in improving the circadian rhythm disturbance in sleep-deprived rats.

Objective:To investigate the effects of naringin(NRG)on apoptosis,proliferation,migration and invasion of oral squa-mous cell carcinoma(OSCC)cells.Methods:NRG of 0,5,10,15,20,25 and 30 μmol/L was respectively used to treat OSCC CAL-27 cells,and CCK-8 assay was used to cell vitality.CAL-27 cells were divided into low,medium and high dose NRG groups(NRG-L,NRG-M and NRG-H),Compound C(AMPK inhibitor)group,NRG-H+Compound C group,and control group(NC group,normal cul-ture).Cell proliferation,apoptosis,migration and invasion were detected by CCK-8 assay,EdU staining,flow cytometry,scratch and Tr-answell assays.The expression of aspartate specific cysteine proteinase-3(Caspase-3),proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),MMP-9,p-AMPK,sirtuin 1(SIRT1)and acetylated NF-κB p65(Ac-NF-κB p65)was detected by Western blot.Results:5,10 and 20 μmol/L NRG was selected as the low,medium and high dose for the treatment of CAL-27 cells,re-spectively.Compared with NC group,in NRG-L,NRG-M and NRG-H groups EdU positive rate,scratch healing rate,A450 value,num-ber of invasive cells,the protein expression of PCNA,MMP-2,MMP-9 and Ac-NF-κB p65 in CAL-27 cells decreased,the apoptosis rate,and the protein of Caspase-3,p-AMPK,and SIRT1 increased(P＜0.05);in Compound C group EdU positive rate,scratch healing rate,A450 value,the number of invasive cells,the protein expression of PCNA,MMP-2,MMP-9 and Ac-NF-κB p65 in CAL-27 cells in-creased,apoptosis rate and the protein expression of Caspase-3,p-AMPK and SIRT1 decreased(P＜0.05).Compound C reversed the effects of high dose NRG on the proliferation,migration,apoptosis and invasion of CAL-27 cells.Conclusion:The inhibitory effects of NRG on proliferation,migration and invasion of CAL-27 cells and the promotion of apoptosis may be related to activation of AMPK and inhibition of NF-κB pathway.

Objective To establish a dynamic prediction model of fatal massive hemorrhage in trauma based on the vital signs time series data and machine learning algorithms.Methods Retrospectively analyze the vital signs time series data of 7522 patients with trauma in the Medical Information Mart for Intensive Care-Ⅳ(MIMIC-Ⅳ)database from 2008 to 2019.According to the occurrence of posttraumatic fatal massive hemorrhage,the patients were divided into two groups:fatal massive hemorrhage group(n=283)and non-fatal massive hemorrhage group(n=7239).Six machine learning algorithms,including logistic regression(LR),support vector machine(SVM),random forests(RF),adaptive boosting(AdaBoost),gated recurrent unit(GRU),and GRU-D were used to develop a dynamic prediction models of fatal massive hemorrhage in trauma.The probability of fatal massive hemorrhage in the following 1,2,and 3 h was dynamically predicted.The performance of the models was evaluated by accuracy,sensitivity,specificity,positive predictive value,negative predictive value,Youden index,and area under receiver operating characteristic curve(AUC).The models were externally validated based on the trauma database of the Chinese PLA General Hospital.Results In the MIMIC-Ⅳ database,the set of dynamic prediction models based on the GRU-D algorithm was the best.The AUC for predicting fatal major bleeding in the next 1,2,and 3 h were 0.946±0.029,0.940±0.032,and 0.943±0.034,respectively,and there was no significant difference(P=0.905).In the trauma dataset,GRU-D model achieved the best external validation effect.The AUC for predicting fatal major bleeding in the next 1,2,and 3 h were 0.779±0.013,0.780±0.008,and 0.778±0.009,respectively,and there was no significant difference(P=0.181).This set of models was deployed in a public web calculator and hospital emergency department information system,which is convenient for the public and medical staff to use and validate the model.Conclusion A set of dynamic prediction models has been successfully developed and validated,which is greatly significant for the early diagnosis and dynamic prediction of fatal massive hemorrhage in trauma.

Objective To investigate the effects of Chaihuang Yishen Granules on renal fibrosis in unilateral ureteral obstruction(UUO)mice and its underlying mechanisms.Methods Twenty-four 8-week-old male C57BL/6 mice were randomly divided into control group,model group,and low and high dose groups of Chaihuang Yishen Granules(6 in each group).In control group,only right kidney ureter was exposed and dissected.In model group,the UUO animal model was established by UUO.In low and high dose groups,mice were administered intragastrically at doses of 3.8 and 7.6 g/kg of Chaihuang Yishen Granules respectively,following the model group's method to establish the UUO model.After 7 days,the mice were euthanized and renal samples were collected.HE and Masson staining were used to observe pathological changes and fibrosis degree of the kidneys in each group,Sirius red staining was used to observe collagen deposition.The expression levels of α-smooth muscle actin(α-SMA),fibronectin(FN),type Ⅰ collagen(Col-Ⅰ),glycogen synthase kinase 3β(GSK-3β),and β-catenin related proteins were detected using Western blotting.Changes in A33 and GSK-3β,β-catenin mRNA levels were measured by RT-PCR.Additionally,a normal transformed C3H mouse kidney-1(TCMK1)was used as control(normal group);an in vitro fibrosis model was established using TCMK1 stimulated with Transforming Growth Factor-β(TGF-β);and an in vitro drug model was established using TCMK1 treated with serum containing Chaihuang Yishen Granules.A33 was overexpressed in TCMK1 cells using a transfection with an A33 overexpression plasmid,and changes in fibrosis-related indicators and the expression of A33 and GSK-3β,β-catenin mRNA were observed.Results RT-PCR results showed that,compared with control group,A33 level was significantly increased in model group,while it was significantly reduced in both low and high dose groups of Chaihuang Yishen Granules(P＜0.05).Western blotting showed that the expression levels of fibrosis-related factors such as α-SMA,FN,Col-Ⅰ in model group were significantly higher than those in control group(P＜0.05);while compared with model group,the expression levels of α-SMA,FN,Col-Ⅰ in low and high dose groups of Chaihuang Yishen Granules were significantly lower(P＜0.05).HE,Masson,immunohistochemical staining results showed that model group had severe kidney structural damage,significant increase in collagen deposition,and significantly higher expression levels of GSK-3β and β-catenin proteins compared with those in control group(P＜0.01).In contrast,low and high dose groups of Chaihuang Yishen Granules had good kidney structure,significant improvement in kidney damage and fibrosis,and significantly lower expression levels of GSK-3β and β-catenin proteins compared with those in model group(P＜0.05).In vitro experiment results confirmed that,compared with normal group,A33 overexpression promoted the upregulation of fibrosis-related factors in TCMK1 cells,significantly increase the expression of downstream target genes GSK-3β and β-catenin mRNA in the Wnt/β-catenin signaling pathway(P＜0.05),and A33 overexpression reversed the cellular fibrosis changes downregulated by the serum containing Chaihuang Yishen Granules(P＜0.01).Conclusion Chaihuang Yishen Granules significantly improve renal fibrosis in UUO mice by downregulating the A33/Wnt/β-catenin signaling pathway,suggesting that A33 may be a potential therapeutic target for renal fibrosis.

Objective To analyze the pharmacodynamic interaction of esketamine and propofol in hysteroscopic surgery by response surface method.Methods Forty-five patients underwent elective hystero-scopic surgery,aged 18.5-64.0 years,BMI 18.5-28.0 kg/m2,ASA physical status Ⅰ or Ⅱ.Compound propofol target control infusion of esketamine(0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,and 0.8 μg/ml)with different plasma drug concentrations were selected to keep the plasma drug concentration of esketamine unchanged,and the plasma drug concentration of propofol was increased step by step.To evaluate body re-sponse caused by dilation of the cervix.A response surface model was used to analyze the pharmacodynamic interaction of esketamine and propofol.Results The three-dimensional response surface of esketamine(0.0-0.8 μg/ml)and propofol(1.0-7.0 μg/ml)showed that the two have an additive effect in sedation and inhibition of body activity reaction caused by dilated cervix.The median effective concentration(EC50)of esketamine was 0.61 μg/ml(95％CI 0.41-0.81 μg/ml),and the EC50 of propofol was 4.69 μg/ml(95％CI 3.17-6.21 μg/ml)when inhibits the body activity reaction caused by dilated cervix.Conclusion Response surface method can qualitatively and quantitatively analyze the pharmacodynamic interaction of es-ketamine and propofol.Esketamine and propofol have additive effects in inhibiting the body activity reaction caused by dilated cervix.