ObjectiveTo analyze the pharmacodynamic substance basis of Shangkeling Spray and its potential mechanisms in intervening knee osteoarthritis （KOA） using ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS）， network pharmacology， and molecular docking technology. MethodsUPLC-MS was used to identify the chemical components of Shangkeling Spray. Pharmacokinetic properties were employed to screen potential active ingredients. Network pharmacology methods were utilized to collect potential targets of these ingredients and the pathological gene set of KOA. An "active ingredient-disease" target network was constructed using databases such as STRING. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） functional enrichment analyses were performed using clusterProfiler. Libraries including NumPy were employed to calculate shortest path lengths to identify dominant pharmacodynamic links. Core gene clusters were identified using MCODE， validated through the Gene Expression Omnibus （GEO） database， and molecular docking was performed between key active ingredients and core targets. ResultsA total of 322 and 314 chemical components were identified under positive and negative ion modes， respectively， with 410 components in total after de-duplication， mainly including flavonoids， coumarins， terpenoids， organic acids， and alkaloids. Analysis of the "active ingredient-disease" network identified "development and regeneration"， "cell growth and death"， "immune system"， and "nervous system" as the dominant pharmacodynamic links of Shangkeling Spray in the treatment of KOA. Molecular docking showed that key active ingredients， such as bletillin A， formononetin， morin， oxymatrine， aconitine， gallic acid， curdione， apigenin， naringenin， and oleanolic acid， tightly bound to functional domains of 10 key targets including Jun proteins（JUN）， interleukin-6 （IL-6）， protein kinase B1 （Akt1）， Caspase-3， nuclear transcription factor-κB subunit p65（RELA）， nuclear factor-kappaB1（NF-κB1）， Cyclin D₁， mammalian target of rapamycin（mTOR）， tumor necrosis factor （TNF）， and Fos proto-oncogene protein （FOS）. These interactions synergistically regulated the phosphatidylinositol 3-kinase （PI3K）/Akt/mTOR-related signaling axis and nervous system-related pathways， mediating cartilage repair， reducing inflammation and pain， and improving KOA. ConclusionThis study preliminarily clarifies the pharmacodynamic substance basis of Shangkeling Spray and suggests that its main active ingredients may improve KOA by synergistically regulating the PI3K/Akt/mTOR-related pathways， providing a reference for subsequent exploration of its substance benchmark and mechanism of action.

Objective To investigate the detection rate of Klebsiella pneumoniae(KP)in the intestinal tract of intensive care unit(ICU)patients and community healthy adults,and analyze its virulence genes and drug resistance.Methods Fecal swabs or fecal specimens from ICU patients admitted to the Third Affiliated Hospital of Wenzhou Medical University and community healthy adults from December 2020 to December 2021 were collected for screening KP.The hypermucoviscosity(HM)phenotype was identified by the wire drawing test.The Vitek 2 automatic microbial analysis system and K-B disk diffusion method were used for detecting drug sus-ceptibility.PCR was used to screen extended spectrum β-lactamases(ESBLs),carbapenem resistance genes,9 virulence related genes,including allS,entB,fimH,iutA,kfu,magA,mrkD,rmpA,and ybtS,and 6 highly virulent capsule serotypes such as K1,K2,K5,K20,K54,and K57.Meanwhile,their multilocus sequence typing(MLST)was analyzed.Statistical analysis was conducted using SPSS 23.0 software.Results A total of 448 fecal samples were collected,including 140 from ICU patients and 308 from commu-nity healthy adults.A total of 89 strains of KP were isolated,including 36(25.7%)from ICU patients and 53(17.2%)from commu-nity healthy adults.The detection rate of KP in ICU patients was significantly higher than that in community healthy adults(χ2=4.375,P＜0.05).There were 9 strains(25%)of HMKP in ICU patients and 19 strains(35.8%)in community healthy adults,and there was no significant difference in the detection rate between them(χ2=1.170,P＞0.05).HMKP and KP detected in healthy adults were natu-rally resistant to ampicillin and still highly sensitive to commonly used antibiotics.The KP from ICU patients showed varying degrees of resistance to commonly used drugs.The resistance of the KP from ICU patients to cephalosporins,penicillinase inhibitors,carbapenem,quinolones,amtronam,gentamicin,amicacin and cotrimoxazole except for tobramycin was significantly higher than that from healthy a-dults(all P＜0.05).In addition,18 strains(50%)of KP producing ESBLs were detected,all of which were non-HM phenotypes.A-mong them,6 strains were carbapenem resistant KP(CRKP).Five strains of CRKP carrying KPC enzyme all belonged to ST11 type and 1 carrying NDM enzyme to ST1855 type.A total of 27(30.3%)strains of KP with highly virulent capsule serotypes were detected,mainly K1,K2,and K57,but no K5.There were no significant differences in the detection rates of 9 virulence genes and 5 highly vir-ulent capsule serotypes between ICU patients and healthy adults(all P＞0.05).High-risk ST types were detected in both ICU patients and healthy adults.Conclusion High virulence and high-risk KP strains are detected from the intestinal tract of healthy adults in this area,which increases the risk of infection transferring from community to hospital.In addition,carbapenemase-resistant ST11 is detec-ted from the intestinal tract of ICU patients,so the spread of drug-resistant plasmids should be highly vigilant.

Objective:To explore the correlation between long non-coding RNA LOC101927476 (LncRNA LOC101927476), stage-specific embryonic antigen-4 (SSEA-4), and intronic microRNA-28 (hsa-miR-28) and postoperative recurrence/metastasis of ovarian cancer.Methods:A total of 195 patients with ovarian cancer who underwent surgical treatment in The First People’s Hospital of Shangqiu and The First Affiliated Hospital of Zhengzhou University from Jan. 2021 to Oct. 2022 were selected. Patients were divided into occurrence group and non-occurrence group according to whether they had recurrence/metastasis within 2 years after surgery. R package "Match It" and the 1∶1 principle for propensity score matching (PSM) were used to compared the expression of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 in different tissues and two groups. Multivariate logistic regression analysis was used to analyze the correlation between various detection indicators in cancer tissues and postoperative recurrence/metastasis of ovarian cancer. The value of LncRNA LOC101927476, SSEA-4 mRNA, hsa-miR-28, and their combined use in predicting recurrence/metastasis in cancer tissues was analyzed using the receiver operating characteristic (ROC) curve. The external calibration curve was used to analyze the combined predicts of the consistency between the incidence of recurrence/metastasis and the actual incidence.Results:In cancer tissues, the expression of LncRNA LOC101927476 and hsa-miR-28 was lower than that in adjacent tissues, while the expression of SSEA-4 mRNA was higher ( P<0.05). The expression of LncRNA LOC101927476 and hsa-miR-28 in the occurrence group was lower than that in the non-occurrence group, while the expression of SSEA-4 mRNA was higher than that in the non-occurrence group ( P<0.05). Multivariate logistic regression analysis showed that the increase of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 were independent factors associated with the recurrence/metastasis of ovarian cancer after surgery ( P<0.05). ROC analysis showed that the AUCs of LncRNA LOC101927476, SSEA-4 mRNA, hsa-miR-28, and their combined prediction of ovarian cancer recurrence/metastasis after surgery were 0.730, 0.767, 0.832, and 0.915, respectively ( P<0.001). Comparing the AUC of the combination with that of the individual, it was found that the AUC of the combination was significantly higher than that of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 ( Z=3.924, 2.995, 2.078, P=0.000, 0.003, 0.038). The calibration curve of the external dataset showed that the combined prediction of the incidence of recurrence/metastasis was basically consistent with the actual incidence, and the two curves had a high degree of fit. Conclusions:The expression of LncRNA LOC101927476, SSEA-4, and hsa-miR-28 in cancer tissues is associated with postoperative recurrence/metastasis of ovarian cancer, which can provide a reference for early clinical prediction of recurrence/metastasis. The combined application of the three can further improve the predictive value, help to early warn the risk of recurrence/metastasis, and provide important reference information for clinical personalized prevention intervention.

Objective:An in vivo mammalian hepatocyte Unscheduled DNA Synthesis(UDS)test was used to evaluate the genotoxicity of Cross-linked Sodium Hyaluronate Gel and Bone Repair Materials,providing experimental evidence for establishing a UDS testing method for medical devices and materials.Methods:0.9％sodium chloride injection and cottonseed oil were used as the solvent for test materials and negative control,respectively.N-dimethylnitrosamine(NDMA)was used as the positive control for the early sampling times,and 2-acetylaminofluorene(2-AAF)was used as the positive control for the late sampling times.SD rats were administered a single dose for toxic exposure,and liver tissues were collected at 4 h and 16 h,respectively.Hepatocytes were isolated using collagenase perfusion.After labeling with 5-ethynyl-2'-deoxyuridine(EdU),and the net average fluorescence intensity(NAFI)of cell nuclei and nucleoplasm was measured by fluorescence microscope.Data from 50 cells were used to analyze the DNA repair level.Results:Compared with the negative control groups,the positive control groups(NDMA and 2-AAF)showed highly statistically significant differences in NAFI(P＜0.01),indicating successful induction of DNA damage.There was no statistically significant differences between the cross-linked sodium hyaluronate gel groups,bone repair material groups and the negative control group(P＞0.05),suggesting that these materials did not significantly induce DNA damage under the experimental conditions.Conclusion:This study first applied EdU labeling technology to the in vivo hepatic UDS assay,achieving non-radioactive labeling through click chemistry reactions.Under the conditions of this study,cross-linked sodium hyaluronate gel and bone repair materials did not exhibit genotoxicity.In the follow-up,the sample range can be expanded and the observation period can be prolonged to further improve the genotoxicity evaluation system of medical devices.

Objective To evaluate the subchronic systemic toxicity of disposable plasma virus-inactivated blood transfusion sets using hydroxyethyl starch(HES)130/0.4 sodium chloride injection as an extraction medium.Methods Firstly,40 Sprague Dawley(SD)rats including 20 male and 20 female ones were seleted and randomly enrolled into a sample group and a control group by sex,with 20 ones in each group.Secondly,instead of plasma HES 130/0.4 sodium chloride injection was used to leach disposable plasma virus-inactivated blood transfusion sets to prepare the test solution by simulating clinical application such as lighting,adsorption and filtration and storage.Finally,the test solution and HES 130/0.4 sodium chloride injection were injected into the tail vein of the SD rats at a dose of 20 mL/kg for 28 d in the sample group and in the control group respectively,and the subchronic systemic toxicity of disposable plasma virus-inactivated blood transfusion sets and the feasibility of using HES 130/0.4 sodium chloride injection as the extraction medium to assess their subchronic systemic toxicity were evaluated with clinical observation,body mass monitoring,clinical pathology examination,gross necropsy and histopathology examination.Results The sample group and control group had no significant differences in mortality rates,clinical observation results,body mass,gross necropsy results,hematological and coagulation examination results and organ weight(all P>0.05);blood biochemical examinations showed the male rats in the sample group had the cholesterol(CHO)values higher while the creatinine(CR)values lower than those in the control group,with the differences being statistically significant(both P<0.05)and the two indexes within the range of the laboratory's historical reference data,and other blood biochemical indexes were not significantly different(all P>0.05);the sample group had the spleen weight-to-body mass ratios of the female rates lower significantly than those in the control group(P<0.05),and the ratios of other organ weight to body mass had significant differences(all P>0.05);histopathology examination showed slight pathological changes in liver,spleen and kidney of female rats and in spleen and kidney of male rats in the sample group,and the female and male rats in the control group had similar pathological changes found in the sample group,which might be caused by HES metabolites.Conclusion Disposable plasma virus-inactivated blood transfusion sets prove to have no significant subchronic systemic toxicity,and its feasible to use HES 130/0.4 sodium chloride injection as the extraction medium to evaluate the subchronic systemic toxicity of disposable plasma virus-inactivated blood transfusion sets.[Chinese Medical Equipment Journal,2025,46(10):29-35]

Objective To investigate the relationship between the expression levels of serum integrin αMβ2(ITG αMβ2)and gasdermin D(GSDMD)in patients with severe acute pancreatitis(SAP)complicated with acute respiratory distress syndrome(ARDS)and the severity of the disease and its value in predicting prognosis.Methods A total of 147 patients with SAP complicated with ARDS(ARDS group)admitted to the Department of Intensive Care Medicine of the Southwest Jiaotong University Affiliated Hospital(Chengdu Third People's Hospital)from August 2021 to October 2023 were selected.According to the oxygenation index(OI),they were divided into mild group(n=35),moderate group(n=46)and severe group(n=66).According to the 28-day prognosis,they were divided into death group(n=77)and survival group(n=70).Another 147 SAP patients without ARDS at the same time period were selected(non-ARDS group).The expression levels of serum ITG αMβ2 and GSDMD were detected by enzyme-linked immunosorbent assay.Spearman method was used to analyze the correlation between serum ITG αMβ2,GSDMD expression levels and OI in patients with SAP complicated with ARDS.Multivariate Logistic regression was used to analyze the factors of death in patients with SAP complicated with ARDS.Receiver operating characteristic(ROC)curve was used to analyze the value of serum ITG αMβ2,GSDMD expression levels in evaluating the death of patients with SAP complicated with ARDS.Results Compared with the non-ARDS group,the expression levels of serum ITG αMβ2(31.95±8.17 ng/L vs 53.33±12.22 ng/L)and GSDMD(2.25±0.55 ng/ml vs 4.39±1.18 ng/ml)in the ARDS group were increased,and the differences were statistically significant(t=17.637,19.899,all P<0.05).The expression levels of serum ITG αMβ2 and GSDMD in mild group,moderate group and severe group increased in turn,and the differences were statistically significant(F=163.069,194.028,all P<0.05).The expression levels of serum ITG αMβ2 and GSDMD were negatively correlated with OI in patients with SAP complicated with ARDS(r=-0.787,-0.778,all P<0.05).The 28-day mortality rate of 147 SAP patients with ARDS was 52.38%(77/147).Compared with the survival group,the expression levels of serum ITG αMβ2(46.96±10.28 ng/L vs 59.11±10.94 ng/L)and GSDMD(3.74±0.98 ng/ml vs 4.98±1.04 ng/ml)in the death group were increased,and the differences were statistically significant(t=6.920,7.415,all P<0.05).The number of extrapulmonary organ failure≥2,prolonged mechanical ventilation time,increased acute physiological and chronic health assessment II score,and increased ITG αMβ2.and GSDMD were independent risk factors for death in SAP patients complicated with ARDS(Wald χ2=4.297～13.536,all P<0.05),and increased OI was an independent protective factor(Wald χ2=8.346,P<0.05).The combined evaluation AUC of serum ITG αMβ2 and GSDMD expression levels results in a larger area under the curve(AUC)for mortality in SAP patients with ARDS compared to the individual evaluation of SAP complicated with ARDS,which was greater than serum ITG αMβ2 and GSDMD expression levels alone,and the differenes were statistically significant(Z=3.517,3.430,all P<0.05).Conclusion The increase of serum ITG αMβ2 and GSDMD expression levels is related to the progression and poor prognosis of patients with SAP complicated with ARDS.The combined monitoring of serum ITG αMβ2 and GSDMD expression levels has a high evaluation value for the risk of death in patients with SAP complicated with ARDS.

Objective:This study employs a combination of single-cell sequencing and Mendelian randomization to explore the genetic associations and molecular mechanisms of Eomes in RCC.Methods:In this study,single-cell transcriptomic data from RCC tissues and adjacent normal tissues were extracted from the GEO database.The data were analyzed using R language and various packages such as Seurat,limma,and CellChat for cell cluster annotation,intercellular communication analysis,and differential expression analysis.Additionally,eQTL data related to differentially expressed genes were retrieved from the GWAS database as exposure variables,with RCC used as the outcome variable in Mendelian randomization analysis to identify the role of Eomes in RCC.Finally,GO functional enrichment and KEGG pathway analyses were conducted to explore the potential molecular mechanisms of Eomes.Results:Single-cell RNA sequencing revealed that B cells play a significant role in the heterogeneity of RCC.Mendelian randomization analysis indicated that Eomes is an important risk factor for RCC(P＜0.05).Furthermore,seven highly correlated specific SNPs were identified,including rs 17021298,rs2247056,rs2617170,rs3806624,rs55908509,rs6590334,and rs9420589.GO and KEGG enrichment analyses suggest that Eomes may be involved in early cell fate determination in renal cell carcinoma and participate in the regulation of Th1 and Th2 cell differentiation,HPV infection,and the Notch signaling pathway.Conclusions:This study is the first to combine single-cell sequencing and Mendelian randomization analysis in RCC,confirming a strong positive causal relationship between Eomes and RCC(OR＞1).Our findings offer new insights into the pathogenesis of RCC,suggesting that Eomes could serve as a novel target for early diagnosis and personalized treatment of RCC.

Aim To investigate the effect of Zhenwu decoction on inflammation,oxidative stress and apopto-sis of human glomerular epithelial cells(HGEC)in-duced by lipopolysaccharide(LPS)based on Nrf2/HO-1 signaling pathway,and to explore the underlying mechanism.Methods HGEC were treated with LPS(1.0 mg·L-1)for 24 h to construct an oxidative damage model.On this basis,2.5％,5％and 10％Zhenwu decoction-containing serum were added to the low,medium and high dose groups of Zhenwu decoc-tion,and a normal group was set up.The changes of cell activity were assessed by MTT method and LDH method.The contents of TNF-α,IL-6,IL-10,SOD,CAT,GSH-Px,ROS and MDA in each group were de-tected by ELISA.The apoptosis of each group was de-tected by flow cytometry.The mRNA and protein ex-pressions of Bax,Bcl-2,caspase-3,caspase-9 and Nrf2/HO-1 pathway were detected by RT-qPCR and Western blot,respectively.Results Compared to the normal group,the model group of HGEC exhibited increased levels of inflammatory cytokines,enhanced oxidative stress response and aggravated apoptosis;after inter-vention with various doses of Zhenwu decoction,the in-flammatory levels in HGEC were reduced,oxidative damage and apoptosis were effectively ameliorated,and the mRNA and protein expression levels of the Nrf2/HO-1 signaling pathway were upregulated.Conclu-sions Zhenwu decoction can protect HGEC from LPS-induced inflammation and oxidative damage and im-prove apoptosis.The mechanism may be related to the activation of Nrf2/HO-1 signaling pathway.

Objective To analyze the specific reasons for the suspension and termination of domestic medical device clinical trial projects.Based on the analysis results,provide reference suggestions and improvement measures for all parties in-volved in the trials.Methods Data collection method was used to collect samples of domestic medical device clinical trials,and visits were made to multiple medical institutions.For projects that were suspended or terminated,specific reasons were obtained through on-site interviews with research teams and clinical trial institution staff,reviewing"Project Suspension/Termination Let-ters"stored in the investigator's folder,and telephone consultations with clinical research associates(CRAs)of the projects.A contract research organization(CRO)commercial company was also visited,and specific reasons for project suspension or termi-nation were obtained through on-site interviews or telephone consultations with project managers,CRA-related personnel,etc.Descriptive analysis was used to summarize the reasons for suspension and termination and their rates.Results Statistical analy-sis showed that the suspension and termination rate of medical device clinical trials in China was 17.30％,with a rate of 16.04％in samples collected from medical institutions and 21.30％in samples collected from CRO companies.The reasons leading to the overall suspension and termination of domestic medical device clinical trials included sponsor strategy adjustments,medical insti-tution or research team issues,product or design defects,switching to registration using data from similar products,trial result-re-lated factors,third-party service provider issues,other reasons,safety-related factors,pandemic reasons,regulatory updates,and difficulties in obtaining informed consent.Conclusion The reasons for the suspension and termination of domestic medical de-vice clinical trial projects are complex and diverse,with a statistical analysis showing a rate of 17.30％in China.

Objective:To explore the correlation between long non-coding RNA LOC101927476 (LncRNA LOC101927476), stage-specific embryonic antigen-4 (SSEA-4), and intronic microRNA-28 (hsa-miR-28) and postoperative recurrence/metastasis of ovarian cancer.Methods:A total of 195 patients with ovarian cancer who underwent surgical treatment in The First People’s Hospital of Shangqiu and The First Affiliated Hospital of Zhengzhou University from Jan. 2021 to Oct. 2022 were selected. Patients were divided into occurrence group and non-occurrence group according to whether they had recurrence/metastasis within 2 years after surgery. R package "Match It" and the 1∶1 principle for propensity score matching (PSM) were used to compared the expression of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 in different tissues and two groups. Multivariate logistic regression analysis was used to analyze the correlation between various detection indicators in cancer tissues and postoperative recurrence/metastasis of ovarian cancer. The value of LncRNA LOC101927476, SSEA-4 mRNA, hsa-miR-28, and their combined use in predicting recurrence/metastasis in cancer tissues was analyzed using the receiver operating characteristic (ROC) curve. The external calibration curve was used to analyze the combined predicts of the consistency between the incidence of recurrence/metastasis and the actual incidence.Results:In cancer tissues, the expression of LncRNA LOC101927476 and hsa-miR-28 was lower than that in adjacent tissues, while the expression of SSEA-4 mRNA was higher ( P<0.05). The expression of LncRNA LOC101927476 and hsa-miR-28 in the occurrence group was lower than that in the non-occurrence group, while the expression of SSEA-4 mRNA was higher than that in the non-occurrence group ( P<0.05). Multivariate logistic regression analysis showed that the increase of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 were independent factors associated with the recurrence/metastasis of ovarian cancer after surgery ( P<0.05). ROC analysis showed that the AUCs of LncRNA LOC101927476, SSEA-4 mRNA, hsa-miR-28, and their combined prediction of ovarian cancer recurrence/metastasis after surgery were 0.730, 0.767, 0.832, and 0.915, respectively ( P<0.001). Comparing the AUC of the combination with that of the individual, it was found that the AUC of the combination was significantly higher than that of LncRNA LOC101927476, SSEA-4 mRNA, and hsa-miR-28 ( Z=3.924, 2.995, 2.078, P=0.000, 0.003, 0.038). The calibration curve of the external dataset showed that the combined prediction of the incidence of recurrence/metastasis was basically consistent with the actual incidence, and the two curves had a high degree of fit. Conclusions:The expression of LncRNA LOC101927476, SSEA-4, and hsa-miR-28 in cancer tissues is associated with postoperative recurrence/metastasis of ovarian cancer, which can provide a reference for early clinical prediction of recurrence/metastasis. The combined application of the three can further improve the predictive value, help to early warn the risk of recurrence/metastasis, and provide important reference information for clinical personalized prevention intervention.