Objective To investigate the mRNA and protein expressions of nine integrin subunits in human keloid-derived mesenchymal stem cells (KD-MSCs).Methods Cultured KD-MSCs and normal skin-derived MSCs (NS-MSCs) served as the experiment group and control group respectively.Real-time quantitative PCR and Western blot were performed to measure the mRNA and protein expressions of nine integrin subunits in the two groups respectively.Statistical analysis was carried out by t test.Results Real-time quantitative PCR showed no significant difference in the mRNA expression level of any of the integrin units α2,α3,α5,αV,α10,α11 or β1 between KD-MSCs and NS-MSCs (all P ＞ 0.05).The mRNA expression level of integrin α8 was decreased,while that of integrin β3 was significantly increased in KD-MSCs compared with NS-MSCs (both P ＜ 0.01).Western blot revealed that the changes in protein expression levels of integrin units α8 and β3 were consistent with those in their mRNA expression levels in both KDMSCs and NS-MSCs (both P ＜ 0.01).Conclusions Integrin units α8 and β3 may be involved in the occurrence and development of keloid,and the receptors made up of them may play important roles in the pathogenesis of keloid.

Objective To investigate the mechanisms of metformin in inducing autophagy of gastric cancer MNK-45 cells.Methods Human gastric cancer MNK-45 cells in logarithmic growth phase were incubated in the culture plates,and were divided into the intervention group [gastric cancer MNK-45 cells were intervened by metformin at different concentrations (2,4,8,16,32,64 mmol/L) for 24,48,72 hours] and the control group (gastric cancer MNK-45 cells were cultured in the DMEM medium).The inhibition rate of gastric cancer MNK-45 cells was detected by MTT method.The IC50 value of metformin on gastric cancer MNK-45 cells was 17 mmol/L.Gastric cancer MNK-45 cells were intervened by metforrnin at 17 mmol/L for 48 hours in the experimental group.Gastric cancer MNK-45 cells in the control group were cultured in DMEM medium at 17 mmol/L for 48 hours.The apoptosis of the gastric cancer MNK-45 cells of the 2 groups were detected by flow cytometry.The mRNA expressions of Bax and Bcl-2 of the 2 groups were detected by RT-PCR.The protein expressions of type Ⅰ LC3b,type Ⅱ LC3b,beclinl,AKT,p-AKT,mTOR,p-mTOR,P70s6k,p-P70s6k of the 2 groups were detected by Western blot.The measurement data were presented as (x) s,and were analyzed using the one-way ANOVA or repeated measures ANOVA.Data of the 2 groups were compared using the t test.Results The inhibition rates of gastric cancer MNK-45 cells were 3.0％ ± 1.1％,8.6％ ± 1.7％,15.9％ ± 1.6％,26.1％ ± 3.4％,37.5％ ± 2.3％,49.7％± 3.6％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 24 hours,5.2％± 1.9％,10.4％±2.1％,26.9％± 1.6％,49.5％± 1.6％,59.1％±2.0％,82.1％±2.2％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 48 hours,and 9.5％ ± 2.2％,17.6％ ± 1.4％,30.6％ ± 2.6％,63.2％ ± 2.6％,78.9％ ± 1.4％,93.3％ ± 2.7％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 72 hours.There were significant differences in the inhibition rates among the 6 groups at the same time points (F =155.174,728.229,743.826,P ＜ 0.05),and significant differences were also observed within the same group at different time points (F =39.420,58.692,166.125,30.383,117.517,311.642,P ＜ 0.05).The apoptosis rates of gastric cancer MNK-45 cells in the experimental group and the control group were 25.4％ ± 1.7％ and 6.9％ ± 0.5％,with significant difference between the 2 groups (t =18.378,P ＜0.05).The relative mRNA expressions of Bax/Bcl-2 mRNA in the experimental group and the control group were 1.88 ± 0.16 and 1.00 ± 0.00,with significant difference between the 2 groups (t =9.743,P ＜ 0.05).The relative protein expressions of LC3 Ⅱ/LC3 Ⅰ and beclin 1 in the gastric cancer MNK-45 cells were 1.65 ± 0.08 and 1.47 ± 0.06 in the experimental group and 0.79 ± 0.03 and 0.56 ± 0.06 in the control group,with significant difference between the 2 groups (t =18.023,18.283,P ＜ 0.05).The relative protein expressions of AKT and P70s6k in the gastric cancer MNK-45 cells were 0.80 ±0.14 and 0.97 t0.21 in the experimental group and 0.96 ±0.17 and 1.37 ±0.23 in the control group,with no significant difference between the 2 groups (t =2.103,1.699,P ＞0.05).The relative protein expressions of mTOR and p-mTOR were 0.58 ± 0.l 1 and 0.57 ±0.15 in the experimental group and 1.88 ±0.23 and 2.36 ±0.25 in the control group,with significant difference between the 2 groups (t =11.293,10.979,P ＜ 0.05).No p-AKT and p-P70s6k expression was detected in the experimental group,and the expressions of p-AKT and p-P70s6k in the control group were 1.00 ± 0.00 and 1.00 ± 0.00,respectively.Conclusions Metformin could induce autophagy,inhibit proliferation and promote apoptosis of gastric cancer MNK-45 cells.The mechanism may be associated with the inhibition of mTOR expression and the expression of mTOR downstream proteins p-P70s6k by mefformin,and then the autophagy of gastric cancer MNK-45 cells happens.

In recent years, tumor is a refractory disease occurring frequently which is the main cause of death. Surgery, radiotherapy and chemotherapy are the usual therapeutic tools. However,radiotherapy and chemotherapy have serious side-effects and surgery can not be used effectively when metastasis happened. Therefore, tumor-targeted therapy has developed as a better way to cure tumor. Development of research on the use of PEG-PLGA nanoparticles as drug carriers are reviewed in this article, furthermore, problems about that are analysed.

Objective Knowing the BRCA gene mutational condition of high risk triple negative breast cancer (TNBC) in Xinjiang Uygur Autonomous and acquiring the differences of clinical and pathologic characteristics between person with BRCA gene mutation and person without it by means of BRCA gene mutation testing for 30 cases of TNBC in Xinjiang Uygur Autonomous.Methods The objects of this study were 30 cases of high risk TNBC from Xinjiang.All the coded sequences of BRCA1/2 gene were amplified by means of extracting genomic DNA from peripheral venous blood.BRCA1/2 gene mutation analysis were prescreened through DHPLC.Then,the result was verified by DNA sequencing.The clinical and pathologic characteristics between person with BRCA gene mutation and person without it of 30 high risk TNBC cases were contrastively analysed.Results In all the 30 cases of BRCA gene mutation testing for TNBC in Xinjiang Uygur Autonomous,there were 5 cases of pathogenic mutations of BRCA gene (5/30,16.7 ％); 4 cases of BRCA 1 mutation (4/30,13.3 ％); 1 case of BRCA 2 mutation (1/30,3.3 ％); and there was no mutation to be found in 25 cases of BRCA gene of TNBC (25/30,83.3 ％).As compared with person without gene mutation,who with it had the characteristics of earlier of TNM,the difference was statistically significant (P =0.040).Conclusion Since the rate of BRCA1 gene mutation of high risk TNBC is higher.It is suggested that the BRCA gene of every patients with high risk TNBC should be tested.Comparing with person with BRCA gene mutation and person without it,there might have differences on clinical pathological characteristics features.Therefor,individualized treatment should be taken into consideration.

Objective To explore the expression of interferon-inducible genes (IFIG) in idiopathic inflammatory myopathies (IIM) patients and its clinical significance.Methods Peripheral blood mononuclear cells from 52 IIM patients and 20 healthy controls were extracted.The transcriptional levels of six IFIG genes:oligoadenylate synthetase 1 (OAS-1),IFN-induced protein with tetratricopeptide repeats 1(IFIT1),IFIT4,interferon-inducible-protein 44 (IFI44),radical S-adenosyl methionine domain containing protein 2 (RSAD2) and myxo-virus resistance gene 1 (MX-1) were detected by real-time polymerase chain reaction (PCR).Interferon scores were calculated and compared.The correlations between interferon scores and clinical features of IIM patients were investigated.Non-parametric Mann-Whitney and Spearman analysis were used for statistical analysis.Results In IIM patients,the transcriptional levels of OAS-1 [90.4(40.2,350.8)],IFIT1 [112.7(49.8,151.1)],IFIT4 [127.8(110.4,200.2)],IFI44 [100.3(39.5,160.7)],RSAD2 [100.9(50.2,559.2)] and MX-1 [110.7(60.3,298.9)] were greatly higher than those in healthy controls [4.5(2.1,27.8),21.5(0.4,31.4)i 21.5(0.8,51.2),5.2(0.4,27.8),27.8(17.5,33.5),25.1(15.7,31.8),respectively] (Z=-5.121,-4.359,-5.301,-4.906,-4.348,-4.151; P＜0.01).Meanwhile,interferon scores were significantly elevated in IIM patients [65.7(3.6,136.9),Z=-5.778,P＜0.01],compared with healthy controls [1.2(0.1,3.1)],especially in those with interstitial lung disease [89.3(41.9,176.7),Z=0.544,P＜0.05].Interferon scores were correlated positively with ESR (r=0.234,P＜0.05).Conclusion The mRNA levels of OAS-1,IFIT1,IFIT4,IFI44,RSAD2 and MX-1 are significantly increased in patients with IIM.Interferon score might be used as a new biomarker for IIM patients.

AIM: To design,prepare and screen out functional small interfering RNA(siRNA) for specifically silencing proliferating cell nuclear antigen(PCNA) gene expression and effectively inhibiting cell proliferation on human hepatoma cell line SMMC-7721,human gastric carcinoma cell line SGC-7901 and human colorectal carcinoma cell line Caco2.METHODS: PCNA siRNA was designed based on previous studies about design guidelines and synthesized in vitro by T7 Mega short script reaction kit according to the manufacturer's instructions.After purification,the integrities of siRNA were identified through 19% denaturing polyacrylamide gel electrophoresis,and the concentrations of the generated siRNA were measured by testing the absorbance at 260 nm using a spectrophotometer.Four synthesized double-strand siRNA were transfected into three types of carcinoma cell lines by LipofectamineTM2000 reagent,respectively.WST-8 assay was employed to examine the proliferative inhibitions of these three cell lines.The subsequent alterations on PCNA mRNA and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochemistry,respectively.RESULTS: 3 sequences,No.2,No.3 and No.4 PCNA siRNA showed effective inhibitions on tumor cells among the 4 candidate siRNA,and a single dose of 50 nmol/L of No.2,No.4 PCNA siRNA showed the most effective inhibitory rates as more than 62% at 48 h after transfection.50 nmol/L of No.2 and No.4 PCNA siRNA transfection caused 72% decrease of PCNA mRNA level and almost completely loss of the protein in human Caco2 cells.CONCLUSION: No.2 and No.4 PCNA siRNA have been screened out in this study,which show the capability to effectively down-regulated PCNA expression and significantly inhibit the proliferation of carcinoma cells.The optimal concentration is 50 nmol/L and satisfactory effects are achieved 48 h after transfection in vitro.

Objective To investigate the expression of P21ras and PIK3CA proteins in hepatitis B virus-related hepatocellular carcinoma(HBV-HCC), post-hepatitis B hepatic cirrhosis (HBV-hepatic cirrhosis)and normal hepatic tissues specimen, and their correlation between HBV-HCC and HBV-hepatic cirrhosis tissues.Methods Using immunohistochemistry, the expression of P21ras and PIK3CA proteins in 34 cases of HBV-HCC, 37 cases of HBV-hepatic cirrhosis and 30 cases of normal liver tissues specimen were detected and compared. Results The mean gray scales of P21ras protein in HBV-HCC, HBV-hepatic cirrhosis and normal hepatic tissue specimen were 138.86 ± 2.9, 145. 34 ± 2.06 and 152.07 ± 1.17 (P ＜ 0. 0l), respectively, and were related to the progression of hepatopathy (P ＜0.01). The mean gray scales of PIK3CA protein in HBV-HCC, HBV-hepatic cirrhosis and normal hepatic tissue specimen were 138.20 ± 3. 14, 149.49 ±0. 78 and 154.71 ± 1.29 (P ＜ 0.01), respectively, and were related to the progression of hepatopathy (P ＜ 0. 01).There were apparent correlation between P21ras and PIK3CA in HBV-HCC and HBV-hepatic cirrhosis respectively (r =0. 64, P ＜0. 05; r =0. 42, P ＜0. 05). Conclusion The overexpression of P21ras and PIK3CA in HCC and hepatic cirrhosis tissue suggests that they participate in the tumorigenesis and development of hepatocellular carcinoma and hepatic cirrhosis, and there may be a signal transduction pathway of P21ras-PI3K in HBV-HCC and HBV-hepatic cirrhosis.

Objective To establish the rat model of type 2 diabetes mellitus (T2DM)-induced Alzheimer’s disease (AD).Methods The rat model of T2DM was established by continuous high fat and high glucose diet in aged rats. Then the rat model was screened and identified by using Morris water maze, cerebrospinal fluid microdialysis technique,ELISA,electrophysiological technique and pathologic method,respectively.Results Compared with the normal group and the T2DM group,the rats in T2DM+DM group had obviously learning and memory impairment;the level ofβ-AP in the hippocampus was significantly higher and the frequency of the spikes induced by Achin the hippocampus was notably lower.Conclusion The rat model of T2DM-induced AD can be successfully established by continuous high fat and high glucose diet in rats.

Diabetic ulcer (DU), one of the common and serious complications in patients with diabetes mellitus, often leads to infection, necrosis and amputation, and has a long and costly treatment period. Because of DU's unclear healing mechanism and the difficulty of delayed healing, its treatment and management have been a major challenge in clinical medicine. In recent years, the potential role of mitochondrial autophagy in DU has become a research hotspot with the in-depth study of mitochondrial autophagy mechanism. Previous studies have shown that mitochondrial autophagy is an important intracellular self-repair mechanism that plays a crucial role in maintaining cellular health and functional stability. During the development of DU, mitochondrial autophagy plays multiple roles in attenuating oxidative stress and inflammatory responses, maintaining mitochondrial functional homeostasis, influencing cell proliferation and repair capacity during DU healing, promoting DU healing, and enhancing antimicrobial capacity. In this paper, we illustrate the multiple roles played by mitochondrial autophagy in DU prevention and treatment, as well as the potential applications of mitochondrial autophagy in DU therapy. It is expected to provide a basis for the clinical application of mitochondrial autophagy in DU treatment, and provide more effective strategies and solutions for the treatment of DU.

Aim To study the effect of ginsenoside F1 on the enzyme activity and expression of gene of CYP3 A4 through activation of pregnane X receptor ( PXR ) . Methods With different concentrations of ginsenoside F1 treated on LS174T cells, the expression of CYP3A4 mRNA was determined by Q-PCR, and the enzyme activity was measured by P450-GloTM CYP3A4 assay according to the manufacturer′s instructions, fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test ginsenoside F1 activates PXR by re-porter gene screening assay. Results The results re-vealed that the levels of CYP3 A4 gene and protein ex-pression were significantly increased by ginsenoside F1 in a concentration-dependent manner. At the same time, reporter gene screening showed that ginsenoside F1 could also enhance the transcriptional activity of PXR. Conclusion Ginsenoside F1 can significantly up-regulate the gene expression and enzyme activity of CYP3A4 via the PXR-CYP3A4 pathway.