OBJECTIVE Toobservetheeffectoftemozolomide(TMZ)incombinationwithtetran-drine(TET)on cell viability,colony formation,migration and cell apoptosis of human glioblastoma U87 cells.METHODS TheviabilityofU87cellstreatedwithTET(8-64μmol·L-1),TMZ(50-400 μmol·L-1 )and TMZ combined with TET (3.2,6.4 μmol·L-1 )was detected by cytotoxicity assays with Cell Counting Kit-8 (CCK-8),the colony formation was detected by Giemsa staining,cell migration ability was detected by Transwell migration assay,cell apoptosis was assayed by flow cytometry using Annexin Ⅴ /PI double staining,and the expression of apoptosis-related proteins expression was detec-tedbyWesternblotting.RESULTS ThedataofCCK-8showedthatTET(r=0.903,P<0.05)orTMZ (r=0.995,P<0.05)could inhibit U87 cell viability alone in a concentration-dependent manner.The cell viability inhibition rate of U87 cells by TMZ co mbined with TET was higher than by TMZ or TET alone. Data showed that the effect of TMZ combined with TET was additive.TMZ 100 μmol·L-1 inhibited U87 cell colony formation and migration ablility compared with normal control.The inhibition rate of U87 cells by TMZ 100 μmol·L-1 combined with TET (3.2 and 6.4 μmol·L-1 )was more significant than by TMZ alone (P<0.05).Compared with TMZ alone,TMZ combined with TET (3.2 and 6.4 μmol·L-1 )signifi-cantly down-regulated the expression of anti-apoptotic protein Bcl-XL,but significantly up-regulated the expression of cleaved caspase 3 protein and cleaved poly(ADP-ribose)polymerase.CONCLUSION TET combined with TMZ can inhibit U87 cell viability,colony formation and migration by activating caspase-dependent apoptotic pathway,resulting in apoptosis.

Objective To explore the association of urinary level of kidney injury molecule-1 (KIM-1) with renal tubulointerstitial lesions in patients with lupus nephritis (LN). And to find a potential clinical biomarker, which could reflect the specific tubulointerstitial lesions of LN. Methods Sixty cases of biopsy proven new-onset LN patients were enrolled into the study. And 10 normal kidney tissues were collected, which came from 10 patients with kidney trauma or renal tumor nephrectomy. Sixty patients with LN were divided into mild disease group, moderate disease group, severe disease group and the extremely severe disease group according to the criteria of 2000 Hill's morphologic index for the evaluation of renal pathology. Urine and renal tissues specimen were collected. The clinical and pathological data were analyzed retrospectively. Urine level of KIM-1 was detected by enzyme linked immunosorbent assay (-). Immunohistochemical staining was used to observe the expression of KIM-1 in the renal tissue. T-test, One-Way analysis of variance(ANOVA) test or rank sum test and Pearson or Spearman correlation analysis were used for statistical analysis. Results ①With the aggravation of tubulointerstitial lesions, the urinary level of KIM-1 was increased gradually, which was shown in the control group [(0.32 ±0.03) ng/ml], mild group [(2.31 ±0.30) ng/ml], moderate group [(6.12 ±0.51) ng/ml], severe group [(12.51 ±1.83) ng/ml] and the extremely severe group [(15.30 ±2.11) ng/ml] (all P<0.05); ② With the aggravation of tubulointerstitial lesions, the expression of KIM-1 in the renal tissue was increased gradually which was shown in the control group [(2.13±0.12)%], mild group [(35.01±0.33)%], moderate group [(51.12± 2.11)%], severe group [(63.50 ±1.80)%] and the extremely severe group [(75.31 ±3.22)%] (all P<0.05);③ Urinary KIM-1 level in LN patients was positively correlated with the expression of KIM-1 in the renal tissue (r=0.870, P<0.01);④Urinary KIM-1 level in LN patients was positively correlated with 24 hUP, systemic lupus erythematosus disease activity index (SLEDAI), renol (R)-SLEDAI, glomerular activity index (GAI), tubulointerstitial ativity imdex (TLAI), chronicity (AI) (r=0.826, 0.741, 0.824, 0.743, 0.871, 0.741, P<0.05), and was negatively correlated with estimated glomerular filtration rate (eGFR) (r=-0.726, P<0.05), while was not correlated with CI (r=0.532, P>0.05). The expression of KIM-1 in the renal tissue was positively correlated with serum creatinine (SCr), quantity of 24 h UP, SLEDAI, R-SLEDAI, GAI, TLAI and AI (r=0.780, 0.780, 0.812, 0.727, 0.735, 0.910, 0.701, P<0.05), and was negatively correlated with eGFR (r=-0.727, P<0.05), while it was no correlated with CI (r=0.543, P>0.05). Conclusion Urinary KIM-1 enables to assess the tubulointerstitial lesion in LN patients, and it can be a sensitive biomarker to predict the tubulointerstitial damage and to evaluate the degree of tubulointerstitial lesions.

OBJECTIVETo investigate the correlation between miR-221 and epithelial-mesenchymal transition (EMT) in drug-resistant glioma cells.

METHODSThe expression levels of miR-221, PTEN, p-Akt, E-cadherin, vimentin, and MRP1 were quantitatively analyzed in Z1 cells (primary drug-resistant cells), Z2 cells (drug-sensitive cells) and Z2-BCNU cells (drug-resistant cells) using fluorescent real-time PCR and Western blotting.

RESULTSThe expression levels of PTEN were significantly increased in Z2 cells compared with Z1 and Z2-BCNU cells which overexpressed miR-221 and vimentin. The expression levels of vimentin, p-Akt and MRP1 were significantly decreased in Z2 cells overexpressing E-cadherin.

CONCLUSIONMiR-221 regulates the expression of EMT-related genes through down-regulation of PTEN and activation of PI3-K/Akt signaling.

Cadherins ; metabolism ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Epithelial-Mesenchymal Transition ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; genetics ; metabolism ; Multidrug Resistance-Associated Proteins ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Vimentin ; metabolism

Objective Calculus Bovis(CB)is a kind of valuable traditional Chinese medicine,which has been used in clinic for a long time.It has been shown to have significant anti-stroke,anti-inflammatory and anti-tumor effects.But its mechanism for treating Prostate cancer(PCa)remains unclear.The purpose of this study was to explore the target and mechanism of its action in the treatment of prostate cancer throμgh network pharmacology and in vitro and in vivo experiments.Methods The effective compounds of Calculus Bovis were collected by TCM pharmacology database and analysis platform(TCMSP).Search for potential compound targets in TCMSP.Search the Drμgbank,GeneCards,OMIM,PharmGkb,and TTD databases for disease targets associated with prost cancer.Disease and compound targets were integrated in the STRING database to construct their interaction network(PPI)to reveal the key targets of compound treatment for prostate cancer.In order to elucidate the mechanism of Calculus Bovis in the treatment of prostate cancer,GO functional enrichment and KEGG pathway enrichment analysis were conducted using Cytoscape software.The mechanism of treating prostate cancer with Calculus Bovis was studied in vitro and in vivo.Results A total of 11 compounds with anti-prostate cancer activity were identified.Oleanolic acid,ursolic acid,ergosterol,deoxycorticosterone,methylcholine and cholverdin were potential effective components.A total of 367 targets of Calculus Bovis compounds and 2152 targets of prostate cancer were found.The core targets of Calculus Bovis in the treatment of prostate cancer included TP53,STAT3,AKT1,HSP90AA1,ESR1,SRC,JUN,RELA,CCND1,CDKN1A,EGFR,AR,etc.The biological functions of Calculus Bovis mainly involve oxidative stress response,response to steroid hormones,cell response to chemical stress,peptide-serine modification and phosphorylation,and protein serine/threonine kinase activity.Calculus Bovis treatment of prostate cancer mainly involves PI3K-AKT signaling pathway,TNF signaling pathway,MAPK signaling pathway,etc.In vitro and in vivo experiments showed that Calculus Bovis promoted apoptosis of PC3 cells of prostate cancer by inhibiting PI3K-AKT signaling pathway.Conclusion Calculus Bovis has a therapeutic effect on prostate cancer,and its function is related to inhibiting PI3K-AKT signaling pathway and promoting apoptosis of cancer cells.

OBJECTIVES: Acute ischemic stroke (AIS) is one of the main causes of disability in middle-aged and elderly people, and early activity plays an important role in functional recovery. This study aims to understand the factors that affect the implementation of early activity in patients with AIS and to provide reference for promoting early activity implementation and developing intervention strategies for AIS patients. METHODS: Using purposive sampling, 19 AIS patients and their caregivers who visited at Stroke Center in the Third Xiangya Hospital of Central South University and the Third Hospital of Changsha from June to December 2021, as well as 19 medical staff, hospital administrators, or community workers providing medical health services to stroke patients, were selected as interviewes. A semi-structured interview was conducted based on the social ecological theory model, and the Colaizzi seven-step method was used to analyze the interview data. RESULTS: According to qualitative interview results, the factors affecting early activity in AIS patients were summarized into 4 themes and 12 sub-themes: medical staff factors (insufficient knowledge and skills, insufficient knowledge of early activity, unclear division of responsibilities), patient factors (severity of the disease, lack of knowledge, psychological pressure, fear of falling), social environmental factors (lack of social support, shortage of human resources and rehabilitation equipment, insufficient medical insurance support), and evidence and norms (the evidence for early activity needs improvement, lack of standardized early activity procedures). CONCLUSIONS Early activity in AIS patients is impacted by factors at multiple levels, including medical staff, patients, social environment, and evidence and norms. Developing comprehensive intervention strategies to address these factors can promote early activity implementation in AIS patients.
Aged ; Middle Aged ; Humans ; Ischemic Stroke ; Accidental Falls ; Fear ; Social Environment ; Stroke