To investigate the detection status of human papillomavirus (HPV) among adult women in high-altitude regions of Xizang.

Objective To investigate the protective effect and mechanism of oleanolic acid(OA)on kidneys in rats with type 2 diabetes mellitus(T2DM).Methods A total of 35 Sprague-Dawley(SD)rats were enrolled in this study.25 SD rats were randomly selected to establish T2DM model,after modeling,20 rats remained and divided into T2DM group(n=6),low-dose oleanolic acid group(LOA,n=6)and high-dose oleanolic acid group(HOA,n=8).And ten rats were selected as normal control group(NC,n=10).The biochemical indicators,24 h urine volume and 24 h urinary microalbumin(UAlb)were compared among the four groups.Renal lipid deposition was evaluated by Oil red O staining.The protein expressions of Adenosine 5'-monophosphate-activated protein kinase(AMPK),p-AMPK and peroxisome proliferator-activated receptor γ coactivator-1 α(PGC-1 α)were detected by Western blot.Results Compared with the NC group,the levels of 24 h urine volume,fasting blood glucose(FPG),serum total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),serum creatinine(Scr),serum uric acid(SUA)and 24 hUAlb were increased(P<0.05),while the body weight,high density lipoprotein cholesterol(HDL-C),p-AMPK and PGC-1α were decrease in the T2DM group(P<0.05).Compared with the T2DM group,the expressions of HDL-C,p-AMPK and PGC-1α were increased(P<0.05),while the levels of 24 h urine volume,FPG,TG,TC and LDL-C,Scr,SUA and 24 hUAlb were decreased in the LOA and HOA groups(P<0.05).Compared with LOA group,the expressions of HDL-C,p-AMPK and PGC-1α were increased(P<0.05),while the levels of 24 h urine volume,FPG,TG,TC,LDL-C,Scr,SUA and 24 h UAlb were decreased in the HOA group(P<0.05).Compared with the NC group,a large number of red-stained lipid droplets were deposited in the renal tubular epithelial cells in the T2DM group.Compared with the T2DM group,the lipid droplet deposition was reduced in the LOA and HOA groups,and the improvement was more significant in the HOA group.Conclusion OA can alleviate renal injury in T2DM rats,which may be linked to activation of AMPK/PGC-1α pathway.

BACKGROUND:Endothelial injury is one of the causes of cardiovascular diseases.Human induced pluripotent stem cells are easy to obtain,have strong differentiation ability,and have less exclusiveness,and their endothelial differentiated cells can be used as ideal cells for cardiovascular disease research. OBJECTIVE:To investigate the effect and mechanism of calycosin on endothelial differentiation of human induced pluripotent stem cells and to provide technical support for microvascular regeneration. METHODS:Human induced pluripotent stem cells were divided into control group and calycosin group(1.25,2.5 μg/mL),and growth factors were added to induce single-layer endothelial differentiation.After the induction of differentiation for 8 days,the positive rate of endothelial cell marker CD144 was detected by flow cytometry.Fluorescent expressions of CD144 and CD31 were detected by the immunofluorescence method.Lentivirus RNAi GFP puromycin was used to silence human-induced pluripotent stem cell Piezo1 mRNA followed by endothelial directed differentiation.After 8 days of differentiation,the positive rate of CD144 in differentiated cells was detected by flow cytometry.The mRNA expression levels of CD144,Piezo1 and MEK were detected by qPCR. RESULTS AND CONCLUSION:(1)Compared with the control group,the positive rate of CD144 was significantly increased in the 1.25 and 2.5 μg/mL calycosin groups(P<0.05).The expressions of CD144,Piezo1,and MEK mRNA were increased in the 2.5 μg/mL calycosin group(P<0.05).The fluorescence expressions of CD144(P<0.01)and CD31(P<0.001)were significantly increased in the 2.5 μg/mL calycosin group.(2)Compared with the shNT group,CD144 positive rate and CD144,Piezo1,MEK mRNA expressions were significantly increased in the shNT + calycosin 1.25,2.5 μg/mL groups(P<0.05).Compared with the shPiezo1 group,the positive rate of CD144 and mRNA expressions of CD144,Piezo1 and MEK had no significant changes in the shPiezo1+calycosin 1.25,2.5 μg/mL groups(P>0.05).(3)It is concluded that 2.5 μg/mL calycosin promotes the differentiation of human-induced pluripotent stem cells into endothelial lineages.Calycosin promotes the downstream MEK expression,thereby promoting the endothelial differentiation of human induced pluripotent stem cells by targeting the expression level of Piezo1.

Objective To detect the expressions of intercellular adhesion molecule-1(ICAM-1)and nu-clear factor(NF)-κB in hepatic tissues of the patients with chronic hepatitis B,and to analyze their correlation with the hepatic inflammatory activity and fibrosis degree.Methods The liver biopsy specimens from 66 pa-tients with hepatitis B and 10 non-hepatopathic controls were selected,and immunohistochemistry and in situ hybridization were used to detect ICAM-1 and NF-κB expression levels in different liver tissues.Results The positive rate of ICAM-1 and NF-κB expression in liver tissues of the patients with chronic hepatitis B was higher than that in normal liver tissues,and the difference was statistically significant(P＜0.05).The expres-sion of ICAM-1 and NF-κB in the patients with hepatitis B was positively correlated with the inflammatory ac-tivity and fibrosis degree(r=0.493,0.496,P＜0.01;r=0.580,0.519,P＜0.01).Conclusion ICAM-1 and NF-κB in the patients with chronic hepatitis B are highly expressed,which is useful in judging the hepatic in-flammatory activity and fibrosis degree.

Objective To investigate the effects of probiotics combined with dietary intervention on pe-ripheral blood glucose and lipid metabolism indicators,placental tissue insulin signaling pathway proteins ex-pression and pregnant outcome in the patients with gestational diabetes mellitus(GDM).Methods A total of 83 patients with GDM in this hospital from December 2021 to December 2022 were selected as the study sub-jects and divided into the probiotics group(probiotics combined with diet intervention,43 cases)and control group(simple diet intervention,40 cases)by the random number table method.The levels of peripheral blood glucose,lipid and insulin resistance related indicators before the intervention and in 8 weeks after the interven-tion were compared between the two groups.The histological changes of placenta were observed by HE stai-ning.The pathological indicators were compared between the two groups.The expression levels of insulin re-ceptor substrate-1(IRS-1),glucose transporter 4(GLUT4)and synaptosome-associated protein of 23 kDa(SNAP23)in placental tissue were detected by immunohistochemistry.The pregnant adverse outcomes were compared between the two groups,and the clinical efficacy of probiotics was evaluated.Results Compared with the control group,the levels of fasting blood glucose(FBG),fasting insulin(FINS),serum triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in 8 weeks after intervention in the probiotics group were significantly decreased(P＜0.05),and the level of serum high density lipoprotein cholesterol(HDL-C)was significantly increased(P＜0.05).There were no significant differences in the incidence rates of poor villi maturation,thickening of dry villi arterioles and capillary filling in villi interstitial between the two groups(P＞0.05).Compared with the control group,the expression levels of IRS-1,GLUT4 and SNAP23 in placen-tal tissue of the probiotics group were significantly increased(P＜0.05).The incidence rates of neonatal hy-poglycemia and neonatal hyperbilirubinemia in the probiotics group were significantly lower than those in the control group(P＜0.05).Conclusion Compared with simple dietary intervention,probiotics combined with dietary intervention has more advantages in improving glucose and lipid metabolism of GDM patients,moreo-ver reduces the adverse events occurrence in newborns.

OBJECTIVE To analyze the use status and development trend of proton pump inhibitors(PPIs) in 33 hospitals in Wuhan, Hubei Province after the implementation of the national centralized drug procurement(NCDP) policy, and to provide reference for promoting the subsequent rational use of NCDP drugs and improving related policies. METHODS To make statistics and analysis of purchasing amount of PPIs, defined daily dose system(DDDs), defined daily dose consumption(DDDc) and utilization rate of 33 hospitals in Wuhan in 2019 and 2022. RESULTS After the implementation of the NCDP policy, the total purchasing amount of PPIs decreased by 53.6%, DDDs decreased by 15.4%, DDDc decreased by 45.2%, and the utilization rate of PPIs injectable dosage forms decreased by 12.6%. After NCDP, the highest growth rate of oral dosage forms was omeprazole(5.7%), followed by rabeprazole(5.0%), while injectable dosage forms showed a significant difference in utilization rate, with a significant decline in NCDP varieties and a significant increase in non-NCDP varieties. The overall NCDP utilization rate of PPIs in Wuhan was 64.9%, with little difference among hospitals of different grades. CONCLUSION The NCDP policy achieves the purpose of reducing the drug cost of patients and improving the accessibility of drugs, and is more optimized in the selection of dosage forms, which is in line with the policy expectations overall; but the quantity and price of PPIs in Wuhan decreased after NCDP, and highlighted a certain tendency in the selection of varieties. In the future, we still need to optimize measures to guide clinical priority in the selection of NCDP drugs, to ensure and improve the implementation of NCDP policy.

Objective To observe the interventional effects of complex probiotic powder on blood bio-chemical indices and vaginal flora in pregnant women with high risk of gestational diabetes mellitus(GDM). Methods One hundred and ten pregnant women with high risk of GDM recruited from the outpatient clinics of the obstetric department and nutrition department in this hospital were randomly divided into the probiotic intervention group (n=55) and the placebo control group (n=55).The intervention group was given 1 bag of complex probiotic powder per day,and the control group was given 1 bag of placebo powder.The colors,shapes,sizes and tastes of powders in the two groups remained the consistency.The intervention duration in this trial was 23 weeks.The questionnaire survey were conducted at baseline and at the intervention endpoint,and the blood biochemical indexes detections were performed at baseline,after 24-28 weeks of gestation,and after 34-38 weeks of gestation,respectively.In addition,the differences in the composition of the vaginal flora structure between the two groups of high risk pregnant women with GDM were detected at the intervention endpoint.Results A total of 97 cases were eventually included in the statistical analysis.Among them,there were 50 cases in the probiotic intervention group and 47 cases in the placebo control group.At the end of the 23 week intervention,there were no statistically significant differences in fasting blood glucose,red blood cells,hemoglobin,leukocytes,ferritin,total protein,albumin,glutamic transaminase,glutamic oxalate transam-inase and total bilirubin between the probiotic intervention group and placebo control group (P>0.05).In terms of vaginal flora,there was a significant difference in some indicators of α diversity between the probiotic group and placebo control group (P<0.05),and there was no significant difference in β diversity between the two groups (P>0.05).At the level of vaginal flora phyla,there were significant differences in the Firmicutes,Actinobacteria,Desulfobacterota,Deferribacterota,and δ-Myxococcota between the two groups (P<0.05).At the level of vaginal flora genera,the relative abundance of Lactobacillus spp.in the probiotic intervention group was significantly higher than that in the control group (P<0.05).Conclusion Complex probiotic pow-der supplementation may have a limited role in improving blood glucose and other related biochemical indica-tors in high-risk pregnant women with GDM.But it has a certain role in regulating the vaginal micro ecological balance in pregnant women with high risk of GDM.

Objective To investigate changes and clinical significance in plasma free amino acid levels during the progression from chronic hepatitis B(CHB)to liver cirrhosis(LC)and ultimately to hepatocellular carcinoma(HCC).Methods The plasma samples of 49 CHB patients,43 CHB-related LC patients and 50 CHB-related HCC patients were analyzed using the Hitachi L-8900 amino acid analyzer.The differences in amino acid levels were compared between groups.Simultaneously,the MetaboAnalyst website was used to analyze the relevant amino acid metabolism pathways.Results A total of 16 amino acids with different expression levels were identified.During the development from CHB to HCC,plasma levels of cystine,phenylalanine and glycine gradually increased,while the ratio of branched chain amino acids/aromatic amino acids gradually decreased.Compared with the CHB and the LC groups,levels of taurine,methionine,tyrosine,ornithine,glutamate,isoleucine and tryptophan were significantly increased in the HCC group,while the level of arginine was significantly lower.Compared with the CHB group,levels of serine,alanine and proline were higher in the LC and the HCC groups,while the level of valine was also higher in the HCC group(all P＜0.05).The metabolic pathway analysis of three groups showed significant changes in various metabolic pathways,including phenylalanine,tyrosine,tryptophan biosynthesis,phenylalanine metabolism,glycine,serine,threonine metabolism,valine,leucine,isoleucine biosynthesis,and taurine metabolism.In addition,by constructing a network diagram of amino acid enzyme gene interactions,a total of 79 metabolic enzyme genes related to amino acid expression were discovered.Conclusion Plasma levels of amino acids are of certain guiding significance for early warning and prognosis of HCC and can provide a theoretical basis and methodological basis for the diagnosis and treatment of HCC in the future.

Objective To investigate the molecular mechanism of ferroptosis in glioblastoma(GBM)and to provide insights for identifying new therapeutic targets.Methods GSE108474 was selected from gene expression omnibus(GEO)database and differentially expressed genes(DEGs)in GBM were obtained by using GEO2R,compared with the gene set in the Ferroptosis database(FerrDb)to identify ferroptosis-related gene.GO and KEGG enrich-ment analyses were conducted using DAVID database.A protein-protein interaction network was created using String website.Hub genes with high connectivity were confirmed using Cytoscape software.Prognostic and immune infiltration analyses were performed using TIMER website.RNA expression levels and gene correlation analyses were carried out using GEPIA website.Differential expression of hub gene proteins was analyzed by using the HPA database.Tumor immune characteristic correlations were examined using TISIDB database.Differences in mRNA expression of hub genes between tumor cells A172 and U251MG and normal astrocytes HA1800 were compared u-sing the quantitative real-time PCR.Results Out of 5 331 differentially expressed genes,114 were related to fer-roptosis.GO and KEGG enrichment analysis suggested that these 114 genes might play roles in positive regulation of gene expression,and affect tumor progression through ferroptosis and autophagy pathways.10 hub genes were i-dentified in the protein-protein interaction network,among which cluster of differentiation 44(CD44),murine double minute 2(MDM2)and signal transducer and activator of transcription 3(STAT3)were found to be highly expressed in tumors with lower survival rates.CD44,MDM2 and STAT3 mRNA expression were higher in GBM cells compared to normal cells.Protein expression of CD44,MDM2 and STAT3 was higher in high-grade glioma tis-sues than that in normal tissues.The expression of three genes in the tumor was negatively correlated with ferropto-sis.Immune infiltration analysis revealed that CD44,MDM2 and STAT3 in the tumor were related to the infiltration of neutrophils,CD4+T cells,and dendritic cells,and the expression of three genes was related to various chemo-kines and their receptors.Conclusion CD44,MDM2 and STAT3 may play a role in tumor ferroptosis and immune regulation,which have the potential to become a therapeutic target for GBM.

Objective To study the changes in erythrocyte osmotic fragility(EOF)in patients with hypercythemia,pol-ycythemia and anemia under a high altitude environment.Methods A total of 174 outpatients,inpatients and people un-dergoing physical examination in Tibet Autonomous Region People′s Hospital in October 2020 were enrolled and divided into four groups according to hemoglobin levels:the polycythemia group(n=42),the physiologic increase group(n=48),the a-nemia group(n=29)and the healthy control group(n=55).They were compared with 67 anemia patients and 67 healthy people in the laboratory of Peking Union Medical College Hospital in the same period as the plain control group.The classic EOF test method was used to compare the results of each group′s EOF.The concentrations of NaCl solution for starting he-molysis(STH)and NaCl solution for complete hemolysis(CTH)were included,and the data were statistically analyzed by SPSS software.Results There was a significant difference in CTH between Zang and Han healthy people(P＜0.05).STH in high altitude polycythemia group(0.52)was higher than that in high altitude healthy control group(0.48)(P＜0.001).Compared with plain,the median CTH of plateau anemia group(0.36)and plateau health control group(0.36)was signifi-cantly higher than that of plain anemia group(0.28)and plain control group(0.28)(P＜0.001).Multivariate logistic regres-sion analysis showed that plateau region,female polycythemia and anemia were risk factors for STH elevation.Conclusion This study finds that the EOF increases in the plateau population and patients with polycythemia,which provides reference for studying red blood cell disorders and blood transfusion strategy in the plateau region.