ObjectiveTo investigate the correlation between neutrophil-to-lymphocyte ratio （NLR）， platelet-to-lymphocyte ratio （PLR）， systemic immune-inflammation index （SII） and the severity of intrauterine adhesions （IUA）. MethodsThe retrospective study included 380 patients who underwent transcervical resection of adhesions （TCRA） from December 2019 to March 2025. Based on the American Fertility Society （AFS） classification， patients were divided into mild （n=61）， moderate （n=225）， and severe （n=94） groups. NLR， PLR， and SII were calculated from preoperative blood tests. Statistical analyses included Kruskal-Wallis test and ordinal Logistic regression. ResultsNLR， PLR， and SII were significantly higher in the severe IUA group compared to the mild group （P<0.05）， with SII showing the strongest predictive ability （OR=1.004， P=0.001）. The number of intrauterine procedures was an independent risk factor （OR=1.27/level， P=0.016）. The predictive model ［Logit（P）=-0.676+0.241×operation times+0.004×SII］ effectively identified severe IUA cases. ConclusionInflammatory markers （particularly SII） are correlated with IUA severity and may serve as non-invasive tools for clinical assessment.

ObjectiveTo investigate the role of Ras-GTPase-activating protein SH3 domain-binding protein 2 （G3BP2） in regulating the activation， proliferation， and migration of hepatic stellate cells （HSCs）. MethodsThe mouse HSCs （JS-1 cell line） were treated with 5 μg/L transforming growth factor-beta 1（TGF-β1） for 24 hours to establish an HSC activation and proliferation model. A G3BP2 knockdown system was constructed using siRNA interference technology. The experiment was divided into four groups： Control， TGF-β1 treatment， TGF-β1+si-NC， and TGF-β1+ G3BP2-siRNA. The expression levels of key fibrosis indicators， including type I collagen （Collagen I）， α-smooth muscle actin （α-SMA）， and G3BP2， were detected by Western blot and RT-qPCR. Cell proliferation activity was assessed using the CCK-8 proliferation assay kit and EdU fluorescence labeling technology. Cell migration ability was analyzed by scratch wound healing assay and Transwell migration assay. The formation level of stress granules was quantified by immunofluorescence microscopy to investigate the effects of G3BP2 on stress granule formation in activated HSCs. ResultsStimulation with TGF-β1 upregulated the expression of G3BP2 in JS-1 cells （RT-qPCR： P0.000 1； Western blot： P0.000 1）， while a downward trend in its expression was observed in the G3BP2‑silenced group （RT-qPCR： P0.01； Western blot： P0.000 1）. Compared with the control group， the TGF-β1 group exhibited increased protein expression levels of α-SMA and Collagen I （RT-qPCR： both P0.01； Western blot： P0.01 and P0.05， respectively）， concomitant with an increased number of stress granules and enhanced cell proliferation and migration capacity （all P0.001）. The experimental results demonstrated that G3BP2 knockout effectively reversed the aforementioned phenotypes， with the G3BP2-silenced group showing reduced expression of fibrotic markers （all P0.01）， decreased stress granule formation （P0.01）， and reduced cell proliferation and migration capacity （all P0.05）， compared to the negative control group. ConclusionG3BP2 enhances the activation， proliferation， and migration of HSCs by promoting the formation of stress granules， thereby accelerating the pathological progression of liver fibrosis. This suggests that stress granules may serve as important regulators in controlling the activation， proliferation， and migration of HSCs.

AIM:To evaluate the pharmacokinet-ics(PK)and pharmacodynamics(PD)of two recom-binant human insulin injection by euglycemic clamp technology in healthy male subjects after a single subcutaneous injection.METHODS:We con-ducted a randomized,open-label,single dose,two period,crossover study.A total of 24 healthy male subjects were enrolled and randomized to receive single subcutaneous doses(0.2 U/kg)of the investi-gational products every period.The PK and PD characteristics were assessed by euglycemic clamp up to 14 hours after dosing.RESULTS:Euglycemic clamp technique was successfully established.C-peptide levels detected at each time point before and after administration indicated that endoge-nous insulin secretion was inhibited in the two groups after administration.The geometric mean ratio of Cmax and AUC0-tand 90％confidence interval(CI)of test preparation and reference preparation under fasting condition were in the range of 80.00％-125.00％.CONCLUSION:The human insulin produced by KP Biotech demonstrated similarity to the reference preparation Humulin? in PK and PD characteristics in healthy Chinese subjects.

Objective To explore the effect of transcranial direct current stimulation(tDCS)in rehabilitation of swallowing function in stroke patients.Methods A total of 86 stroke patients diagnosed and treated in Affiliated Rehabilitation Hospital of Zhejiang Chinese Medical University from December 2022 to December 2023 were selected as study objects,and they were divided into observation group and control group according to random number table method,with 43 cases in each group.The control group received routine rehabilitation programs,while the observation group received tDCS therapy on the basis of control group.The swallowing function,actual swallowing process and quality of life of two groups were compared before and after intervention.Results After the intervention,standardized swallowing assessment scores of observation group were significantly lower than those of control group(P＜0.05).The scores of oral and pharyngeal phases of observation group were higher than those of control group(P＜0.05),and there was no significant difference in esophageal phase score between two groups(P＞0.05).The quality of life score of observation group was significantly higher than that of control group(P＜0.05).Conclusion tDCS can improve the swallowing function of stroke patients and improve the quality of life,which is worthy of clinical application.

Objective:To evaluate the accuracy of p53 immunohistochemistry for predicting the mutational status of TP53 in gliomas.Methods:A retrospective study was conducted on 242 diffuse gliomas diagnosed at the Department of Pathology, the First Affiliated Hospital of Fujian Medical University, Fuzhou, China from June 2022 to March 2023. All cases underwent next-generation sequencing (NGS) and p53 immunohistochemical staining. The best threshold in the percentage of p53 immunohistochemical expression was assessed as an alternative to testing for TP53 mutation.Results:Among the 242 diffuse gliomas (WHO grade 2-4), 94 cases had a TP53 mutation. The p53 immunohistochemistry results revealed a significantly increased probability of TP53 mutation when the p53 immunohistochemical positivity rate (based on strongly positive cell count) was ≥20% ( P<0.05). The sensitivity and specificity of p53 immunohistochemistry for predicting TP53 gene mutations were 75.6% and 90.4%, respectively. When p53 immunohistochemical stain was totally negative, the probability of TP53 mutation increased significantly, and the mutation ratio of TP53 gene was 6/17 in all negative cases. Conclusions:When the percentage of p53 positive cells (based on strongly positive cell count) in glioma is ≥20%, p53 immunohistochemistry can be used as a reliable alternative method for TP53 mutation detection. When p53 immunohistochemistry is completely negative, the mutation rate of TP53 gene is higher, and further gene sequencing is recommended to determine the mutation status.

BACKGROUND:Allogeneic tendons are often used in clinical treatment of tendon and ligament injuries,which need to be treated before clinical application.At present,allogeneic tendons will face multiple freeze-thaw cycles after cryopreservation,but there is no relevant research on the changes in the properties of the treated allogeneic tendon after several freeze-thaw cycles.OBJECTIVE:To explore the changes in mechanical properties and composition of allogeneic tendon after multiple freeze-thaw cycles.METHODS:The treated allogeneic tendon was frozen and thawed 3,6,and 10 times at-40 ℃(or-80 ℃),respectively.The mechanical test,DNA content,glycosaminoglycan sulfate,hydroxyproline content and total protein content of the allogeneic tendon after freezing and thawing were detected.RESULTS AND CONCLUSION:(1)There was no significant difference in the maximum load,maximum load elongation,tensile strength and elastic modulus between groups after freezing-thawing cycles for 3,6,and 10 times at-40 ℃(P＞0.05).There was no significant difference in the maximum load and maximum load elongation between the groups after freezing-thawing cycles for 3,6,and 10 times at-80 ℃(P＞0.05).The tensile strength of the group with 3 cycles of freezing and thawing was higher than that of the groups with 6 and 10 cycles of freezing and thawing(P＜0.05).The elastic modulus of the group with 3 cycles of freezing and thawing was higher than that of the group with 10 cycles of freezing and thawing(P＜0.05).(2)The freezing and thawing times did not affect the contents of DNA,hydroxyproline,total protein,and sulfated glycosaminoglycan in allogeneic tendon when stored at-40 ℃(or-80 ℃).(3)These findings suggest that in-40℃ storage,the performance of the allogeneic tendon should not be affected by taking out and putting back the tendon less than 10 times,while in-80 ℃ storage,the allogeneic tendon should be taken out and put back less than 6 times.

Objective To explore the effect of transcranial direct current stimulation(tDCS)in rehabilitation of swallowing function in stroke patients.Methods A total of 86 stroke patients diagnosed and treated in Affiliated Rehabilitation Hospital of Zhejiang Chinese Medical University from December 2022 to December 2023 were selected as study objects,and they were divided into observation group and control group according to random number table method,with 43 cases in each group.The control group received routine rehabilitation programs,while the observation group received tDCS therapy on the basis of control group.The swallowing function,actual swallowing process and quality of life of two groups were compared before and after intervention.Results After the intervention,standardized swallowing assessment scores of observation group were significantly lower than those of control group(P＜0.05).The scores of oral and pharyngeal phases of observation group were higher than those of control group(P＜0.05),and there was no significant difference in esophageal phase score between two groups(P＞0.05).The quality of life score of observation group was significantly higher than that of control group(P＜0.05).Conclusion tDCS can improve the swallowing function of stroke patients and improve the quality of life,which is worthy of clinical application.

Objective The three UGT genes involved in the synthesis of flavonoid glycosides in the wild chrysanthemum were cloned,fused and expressed in prokaryotic system,which provided a basis for further study of the functions of these genes in the synthesis of flavonoid compounds.Methods three candidate UDP-glycosyltransferase(UGT)family genes were successfully cloned from flower tissue cDNA of diploid Chrysanthemum indicum.L in Hubei,namely:CiUGT1(1425 bp),CiUGT2(1311 bp)and CiUGT3(1314 bp)encoded opal fraction of 51.72,48.08 and 48.28 kDa,respectively.After the physical and chemical properties of the protein were analyzed,the recombinant plasmid was constructed by seamless Klon technology,and the recombinant proteins of CiUGT2 and CiUGT3 were successfully expressed in Escherichia coli.Results SDS-PAGE analysis showed that the purified CiUGT2 and CiUGT3 protein samples were about 0.2 mg/mL and 1.4 mg/mL,respectively.Conclusion Two candidate UGTs were obtained to catalyze the synthesis of flavonoid 7-O-glucosides in Chrysanthemum indicum.L,which provide the theoretical basis for the molecular mechanism of flavonoid glycosides biosynthesis in the capitulum of Chrysanthemum indicum.L.

Objective To provide scientific basis for layout planning and standardized cultivation the species of Chrysanthemum indicum complex in future periods,the potential ecological suitable areas was analyzed,.Methods By collecting distribution data of 1379 valid C.indicum complex species nationwide and 104 ecological factors,combined with MaxEnt model and ArcGIS software,the potential ecological distribution areas of C.indicum complex under 17 different climate scenarios was analyzed.Results Precipitation and solar radiation are key environmental variables affecting the suitable habitats of C.indicum complex.The most suitable regions for the growth of C.indicum complex was concentrated in Hubei(C.indicum L.),Shanxi(C.lavandulifolium),Jiangsu(C.nankingense),and Hubei(C.indicum var.aromaticum)regions.Under different scenarios,the suitable habitat area for C.indicum L.and C.lavandulifolium showed an overall decreasing trend,while those for C.indicum var.aromaticum and C.nankingense showed an expanding trend.The centroid shift indicates sensitivity of C.indicum complex to climate change.Conclusion The potential distribution areas of C.indicum complex is classified into ecological suitability levels,providing important reference for the sustainable use,introduction cultivation,and scientific zoning of related resources.

BACKGROUND:The imbalance between proliferation and apoptosis of chondrocytes plays an important role in the occurrence and development of osteoarthritis. Previous studies have found that hsa-miR-3202 is involved in regulating the proliferation and apoptosis of various cells. However,no studies have explored the correlation between hsa-miR-3202 and osteoarthritis.OBJECTIVE:To investigate the expression of hsa-miR-3202 in osteoarthritic chondrocytes and its effect on the proliferation and apoptosis of chondrocytes. METHODS:(1) MicroRNAs differentially expressed in osteoarthritic chondrocytes were screened by biogenic analysis. Based on the current research situation at home and abroad,hsa-miR-3202 was selected for follow-up studies,and its target genes were predicted by gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. (2) Human normal chondrocyte cell lines C28/I2 in logarithmic growth phase were selected and randomly divided into four groups for culture:in normal group,cells were cultured in normal medium for 24 hours,the medium was then changed to normal medium for another 6 hours of culture,and changed to normal medium for subsequent culture;in lipopolysaccharide group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,the medium was then changed to normal medium for another 6 hours,and changed to normal medium for subsequent culture;in lipopolysaccharide+NC group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,and then transfected with has-miR-3202 mimics control for 6 hours,and the medium was change to normal medium for subsequent culture;in lipopolysaccharide+hsa-miR-3202 mimics group,cells were cultured in lipopolysaccharide-containing medium for 24 hours and then transfected with has-miR-3202 mimics for 6 hours,and the medium was changed to normal medium for subsequent culture. After further 48 hours of culture,the expression level of hsa-miR-3202 was detected by fluorescence quantitative PCR and cell apoptosis was detected by flow cytometry. After further culture of 0-72 hours,cell proliferation activity was detected by cell counting kit-8. RESULTS AND CONCLUSION:Bioinformatics analysis results indicated that hsa-miR-3202 was significantly down-regulated in osteoarthritic chondrocytes. GO functional enrichment and KEGG pathway enrichment showed that the function of hsa-miR-3202 target gene was closely related to cell growth and apoptosis. The results of in vitro cell experiments showed that compared with the normal group,the expression level of hsa-miR-3202 and proliferation ability of chondrocytes were significantly decreased in the lipopolysaccharide group (P<0.05),while the apoptotic rate was significantly increased (P<0.05). Compared with the lipopolysaccharide group,the expression level of hsa-miR-3202 and proliferation ability of chondrocytes were significantly increased in the lipopolysaccharide+hsa-miR-3202 mimics group (P<0.05),while the apoptotic rate was significantly decreased (P<0.05). To conclude,the expression of hsa-miR-3202 is down-regulated in osteoarthritic chondrocytes to inhibit cell proliferation and promote cell apoptosis,thus affecting the occurrence and development of osteoarthritis.