Objective To investigate the effects of matrine on proliferation and apoptosis of human rhabdomyosarcoma(RMS) RD cells line in vitro,to study the regulatory mechanism of Wnt/β-catenin pathway-influenced apoptosis of RD cells line by detecting the expressions of β-catenin protein,Bcl-2 protein and caspase-3 protein,and to explore Wnt/β-catenin mechanism during the process of RMS.Methods The human RMS RD cells line was treated with matrine of different concentrations (0.5,1.0,1.5,2.0 g/L)for 48 hours respectively,and the proliferation inhibition rates of different concentrations of matrine on RD cells were detected by methyl thiazolyl tetrazolium assay,while the apoptosis rates by flow cytometry (FCM) and the expressions of β-catenin,Bcl-2 and caspase-3 by Western blot.Results The proliferation inhibition rates between control group and different concentrations of matrine groups were(13.70 ±0.25)％,(33.16 ±0.11)％,(42.96 ±0.90)％,(56.26 ±0.79)％ and (67.89 ±0.63)％,respectively.The apoptosis rates were (5.49 ± 0.96) ％,(17.23 ± 5.03) ％,(25.84 ± 4.17) ％,(36.08 ± 3.68) ％and (47.79 ± 4.82) ％,respectively.The highest expression of β-catenin and Bcl-2 proteins and the minimum amount of caspase-3 protein were found in the control group.After intervention of matrine,the expressions of β-catenin and Bcl-2 reduced while the amount of caspase-3 rose significantly,which was concentration-dependent obviously.Differences were found between every concentration of matrine group with control group according to statistics (all P ＜ 0.05).Conclusions Matrine can inhibit proliferation and induce apoptosis of RD cells.Matrine can down-regulate the expression of β-catenin and Bcl-2 proteins in RD cells,while the amount of caspase-3 protein rises.Wnt/β-catenin signal pathway plays an important role in the apoptosis of RD cell induced by matrine,and its downstream proteins Bcl-2 and caspase-3 are also involved in the regulation of this process.

Liver fibrosis is a common pathological consequence of a variety of chronic stimuli, including viral, autoimmune, drug-induced, cholestatic and metabolic diseases. Fibrosis is driven by a dynamic process involving increased synthesis of matrix components and a failure of physiological mechanisms of matrix turnover. Activation of hepatic stellate cells (HSCs) remains a central event in fibrosis. HSCs are the main source of extracellular matrix (ECM). Transforming growth factor-beta (TGF-β), which is the fibrogenic master cytokine, can induce the activation of HSCs to produce a large amount of ECM, and is capable of inducing apoptosis of liver cells. RNA interference (RNAi) is a novel gene disruption technology. Studies have shown that small interfering RNA (siRNA) targeting TGF-β1 may inhibit the activation and proliferation of HSCs, suppress ECM synthesis and block liver fibrosis. TGF-β1 siRNA-mediated gene silencing therapy provides a new avenue for liver fibrosis. This review summarizes recent progresses in research on HSCs, TGF-β1 and TGF-β1 siRNA in liver fibrosis.

Objective To investigate the effects of dexamethasone(DEX)on the secretion of interleukin (IL)-17 and interferon(IFN)-γ and the proportion of Th17,Tc17,Th1 ,Tc1 cells in peripheral blood mononuclear cells(PBMCs)of patients with systemic lupus erythematosus(SLE). Methods Thirty hospitalized SLE patients were recruited and twenty-two healthy volunteers were recruited as healthy controls. PBMCs were separated from SLE patients and healthy controls and then was cultured in vitro by medium or PMA/Ionomycin or PMA/Ionomycin +dexamethasone for six hours. Four- color immunofluorescent staining and flow cytometric assay were used to analyze the percentage of Th17,Tc17,Th1,Tc1 cells in PBMCs. Concentrations of IL-17 and IFN-γ in plasma and the supernatants of PBMCs which were cultured for 24 hours were measured by enzyme linked immunosorbent assay (ELISA). Results The plasma concentrations of IL-17 and IFN-γwere elevated in SLE patients as compared to the controls(P ＜ 0.05). No significant differences were observed between patients and controls for the spontaneous production of IL-17 and IFN-γ or percentage of T subsets expressed by PBMCs. After the stimulation of PMA,compared with the controls,the level of IL-17 was significantly elevated in the supematants of PBMCs and the percentages of Th17 and Tc1 in SLE patients increased significantly(P ＜ 0. 05). However,there showed no significant differences between SLE patients and the controls for the percentages of Th1 and Tc17 cells. DEX could significantly decrease the production of IL-17(P ＜ 0. 01)and the percentages of Th17,Tc1 cells by the active PBMCs(P ＜ 0. 05). Conclusions There is abnormal expression of T subset cells and their cytokines in vivo of SLE patients. DEX can interfere with immunological pathological process in the cytokine network imbalance of SLE patients and shows powerful inhibition of IL - 17. Our results may provide some laboratory evidence for the clinical application of corticosteroids.

Objective To explore the change and the spectra of opportunistic infections in AIDS patients after antiretroviral treatment in He′nan province.Methods Inpatients of post treatment with disease in accordance with the criteria were selected from the Sixth People′s Hospital of Zhengzhou.A retrospective study was carried out, including the content such as gender,age,infection route,treatment time,the date of complication,and CD4+T lympho-cyte counts when the patient with complications.Disease were divided into four group:opportunistic infections,adverse drug reactions,tumor,cardio-cerebrovascular disease and diabetes mellitus.Statistic case number and incidence of every kinds of the disease in the different stages of HAART.Results (1)1 001 cases of patients were involved in the result analysis.People of 41-60 years old,the peasants and peasant workers,accounted for 57.4%and 78.9%.The infection route was mainly through blood transmission and blood collection, accounting for 83.45%.Most of the patients come from Zhumadian, Xuchang, Zhoukou, Nanyang, which were high -incidence area in He′nan.( 2 ) In 1 001 patients of post treatment, the rate of opportunistic infections was 58.4%, while adverse drug reactions was 30.5%.Bacterial pneumonia,extrapulmonary tuberculosis,oral cavity fungal infection and tuberculosis were common in opportunistic infections of patients of post treatment.The main adverse reactions were allergic dermatitis,liver injury and myelosuppression.The liver cancer case was the highest ( 8/29 ) .( 3 ) 1 001 cases were divided into 6 groups according to different count of treatment time,during 3 months of treatment,the incidence rate of opportunistic infec-tions and adverse drug reactions was the highest,accounted for 75.2% and 41.6%.After 5 years of treatment,the incidence rate of cardio-cerebrovascular disease and diabetes mellitus was 6.39%.With the prolonging of HAART time,incidence of opportunistic infections and adverse drug reactions remarkably reduced,but the occurence rate car-dio-cerebrovascular disease and diabetes mellitus increased.Conclusion In AIDS patients after HAART in He′nan at present,opportunistic infections and adverse drug reactions are main diseases and used to occur during three months after HAART.Attention will be focused on tumor, cardio -cerebrovascular disease and diabetes mellitus in AIDS patients of long-term survival.

AIM To investigate the expression of pep-tide transporter (PEPT)2 mRNA in the lungs of ratswith lipopolysaccharide(LPS)-induced acute lung inju-ry. METHODS Health male Sprague-Dawley rats were divided randomly into 5 groups: normal control,cheally) given groups. Lung histopathological changes were observed under light microscope. Lung wet-to-dry weight (W/D) ratios, total protein concentration in bronchoalveolar lavage fluid (BALF) and lung my-eloperoxidase(MPO) activity were measured. The ex-pression levels of PEPT2 mRNA were evaluated by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS Typical pathological inflammation changes in the lungs, including alveolar congestion, hemorrhage, edema, infiltration of neutro-phils in the air space or vessel wall, thickness of the alveolar wall, and hyaline membrane formation, were observed in LPS groups. The lung W/D ratios in LPS 2, 4 and 8 h groups (4.72±0.18, 5.06±0. 17 and 5.12±0.16) were significantly higher than those innormal control and saline groups (4.12±0. 15 and 4.14±0.11). Compared with normal control and sa-g-1 wet tissue], MPO activity increased obviously in LPS2,4 and 8 h groups[(1.96±0.15), (2.23±protein concentration in BALF in LPS 2, 4 and 8 hgroups [ (36.6±2.9), ( 86.9±3.5 ) and (92.2±There was no statistical difference in PEPT2 mRNA levels in the lungs of rats among all groups. CON-CLUSION PEPT2 mRNA level does not change sig-nificantly in the acute injured lung tissue induced by LPS.

OBJECTIVETo investigate the apoptosis-inducing effect of DMF on the human liver cells (HL-7702) in vitro.

METHODSLiver cells were exposed to different concentrations of DMF (0, 50, 100, 150, 200 mmol/L) for 12 hours. Apoptotic rate, the expression of Bax, Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively.

RESULTSThe increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group (P < 0.01), the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance (P > 0.05), as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control (P < 0.01). The new lands of procaspase-3 in 150, 200 mmol/L were observed, which demonstrated that there was active caspase-3.

CONCLUSIONDMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; Dimethylformamide ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Background The hyperplasia of human Tenon capsule fibroblasts (HTFs) is a common cause of filtering surgery failure in glaucomous eye.Researches demonstrated that hydroxycamptothecin is a cell cycle arresting drug and induce apoptosis of cancer and fibroblasts.However,its mechanism is currently less understood.Objective This study was to investigate whether hydroxycamptothecin induce the apoptosis of HTFs and explore the possible mechanism.Methods Human Tenon capsule tissue was obtained from EyeBank of Jiangsu Province Hospital.HTFs were cultured using explant method in vivo and passaged in DMEM containing 10％ FBS.The cells were identified using vimentin and keratin by immunochemistry,and the cells of generation 3-6 in the logarithmic growth phase were used in the experiment.The cells were incubated with 0.01,0.05 or 0.10 g/L hydroxycamptothecin for 5 minutes respectively,and the cells without any hydroxycamptothecin were served as the control group.Cell viability then was assessed by cell counting kit-8 (CCK8) for the optimal inhibition concentration.The cells were treated by 0.10 g/L hydroxycamptothecin for 24 hours,and the apoptotic rate of the cells were assayed with annexin V/PI double-staining.Mitochondrial membrane potential of HTFs was assessed using JC-1 staining.The expressions of caspase-3,caspase-9 and cytochrome C (cyt C) in mitochondria and cytoplasm of HTFs were detected by Western blot.Results The proliferative value (A450) of the HTFs 0,0.01,0.05,0.10 g/L was 0.9716±0.0608,0.8035 ± 0.0346,0.7048 ±0.0446,0.6265 ±0.0286,with a significant difference (F =26.372,P =0.002).A450 of HTFs in the 0.01,0.05,0.10 g/L groups was significantly lower than the control group (P＜0.05),with the lowest A450 value in the 0.10 g/L group.The apoptotic percentage of HTFs was (18.72±1.41)％,in the 0.10 g/L hydroxycamptothecin group and that of the control group was (3.67 ±0.36)％,showing a significant difference between them (t =-10.374,P=0.001).The expression intensity of caspase-3 and caspase-9 protein in HTFs was higher in the 0.10 g/L hydroxycamptothecin group than that in the control group.JC-1 staining showed that the green fluorescence of the monomer JC-1 in cytoplasm was stronger in the 0.10 g/L hydroxycamptothecin group than that in the control group,but the red fluorescence of the polymer JC-1 in the 0.10 g/L hydroxycamptothecin group was weaker than that in the control group.The grey scale of cyt C protein in HTFs in mitochondrion was 0.0605±0.0022 in the 0.10 g/L hydroxycamptothecin group,showing a significant increase in comparison with 0.0301 ±0.0016 of the control group (t=4.865,P=0.014).However,the grey scale of cyt C protein in cytoplasm was declined in the 0.10 g/L hydroxycamptothecin group than that in the control group (0.0605 ±0.0022 vs.0.0301 ±0.0016) (t =-11.177,P =0.001).Conclusions Hydroxycamptothecin can induce the apoptosis of HTFs through activating the mitochondrial apoptosis pathway.

Objective To prove that valve sparing aortic root reconstruction may have an impact on the outcome and longevity of the repair for patients with acute type A aortic dissection.Methods Clinical data of 98 consecutive AAAD patients undergoing the valve sparing aortic root reconstruction were analyzed.The postoperative follow-up and the clinical effect of the procedure were retrospectively analyzed.Results Pre-operative grade of aortic insufficiency was(2.3 ± 0.5), CPB-time was (217.8 ± 43.1)min, aortic cross clamp time was(142.7 ± 37.4)min and stay on ICU (11.1 ± 5.7) days, while hospitalisation was(14.3 ± 3.2) days.8 patients(8.2％) died peri-operatively.None of the early deaths were valve-related.Rethoracotomy rate was 3.1％.All patients followed up(19.3 ± 7.6) (6-68) months.Survival at 1 year was 90.8％.Freedom from valvular reoperation was 97.9％ at 3 years.At last investigation, mean grade of aortic insufficiency for AADA was 0.5 ± 0.3 (0-3).Conclusion Regardless of the underlying indication, the aortic valve preserving reimplantation technique can be performed with favourable functional results.

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen.Studies have shown that VEGF is closely associated with ischemic stroke,and this makes it possible to intervene in ischemic stroke from the level of VEGF and its receptor.This article reviews the biological effect of VEGF and its receptor,mechanism of action involving in various stages of ischemic stroke,and the therapeutic prospect in ischemic stroke.

BACKGROUND:Periprosthetic femoral fracture is a severe complication after total hip arthroplasty. More than 85% belongs to Vancouver B2 and B3 fractures, and the operation is difficult. OBJECTIVE:To explore the clinical effect of Cable-Ready fast system and biotype long-stem prosthesis and its effect on the recovery of joint function in patients with Vancouver B2 and B3 type periprosthetic femoral fractures after total hip arthroplasty. METHODS: A total of 60 patients receiving total hip arthroplasty suffered from Vancouver B2 and B3 type periprosthetic femoral fractures in the Huanggang Central Hospital from September 2011 to September 2012. They were equaly divided into control and observation groups according to different fixation methods. Patients in the control group were treated by ordinary steel wire cerclage fixation combined with uncemented long-stem prosthesis; those in the observation group were treated by Cable-Ready fast system combined with fast uncemented long-stem prosthesis. RESULTS AND CONCLUSION:The average operation time, the time of hospitalization and fracture healing time were shorter in the observation group than in the control group. Moreover, the intraoperative blood loss was less in the observation group than in the control group. After 1 year of folow-up, Harris hip score was higher in the observation group compared with pre-treatment (t=3.174 9,P=0.002 6), and significantly higher than the control group (t=2.479 8,P=0.015 4). The excelent and good rate of Harris hip score was significantly higher in the observation group than in the control group (χ2=11.294 5,P=0.002 6). The total incidence of complications was significantly lower in the observation group than in the control group (χ2=8.139 7,P=0.04 2). These data indicate that Cable-Ready fast system combined with uncemented long-stem prosthesis in the treatment of Vancouver B2 and B3 periprosthetic femoral fractures after total hip arthroplasty has smal injury, less postoperative complications and better recovery of hip function after operation.