objective To assess the effect of aldosterone on the production of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9)and collagen Ⅳ in culture supematants of podocytes and the possible molecular mechanisms involved in the influence of aldosterone on the synthesis and degradation of extracellular matrix produced by podocytes. Methods Podecytes were treated with aldosterone at the concentration of 10-11, 10-9, 10-7 mol/L respectively. Cultured podocytes were examined at 24, 48 and 72 hours respectively. Spironolactone, a receptor antagonist of aldosterone, was added to observe the blocking effect on aldosterone. An inhibitor of TGF-β1 receptor was used to determine whether the effect of aldosterone on podocytes were mediated through TGF-β1 system. The enzymatic activities of MMP-2 and MMP-9 were assayed by gehtin zymography. Collagen Ⅳ 0.5 chain and TGF-β1 proteins released into culture supematants were assessed by Western blot and ELISA analysis. The adhesion rate of podocytes was monitored by flow cytometry. Results Aldosterone increased the activities of MMP-2 and MMP-9 in a dose- and time-dependent manner (P<0.05). Aldosterone decreased the level of collagen Ⅳ or5 chain protein in culture supernatants (P<0.05). Meanwhile, the expression of TGF-β1 was also increased (P<0.05). Spironolactone completely abolished the above-mentioned changes(P< 0.05). Blockage of TGF-β1 signaling with SB431542 prevented the aldosterone-induced upregulation of MMP-2 and MMP-9 as well as the downregulation of the collagen Ⅳ α5 chain protein and the adhesion rate of podocytes (P<0.05). Conclusions Aldosterone increases the activities of MMP-2 and MMP-9 but decreases the expression of collagen Ⅳ α5 chain and the adhension rate of podocytes possibly via TGF-β1 signaling pathway. Such alterations may contribute to glomerular podocyte injury associated with the GBM abnormality caused by the imbalance between matrix synthesis and degradation.

Forty-two patients with type 2 diabetes (DM group),23 type 2 diabetic patients with unstable angina pectoris (DM + UAP group),and 36 healthy subjects (control group) were enrolled in this study.Serum samples were collected for determining serum concentrations of vascular endothelial growth factor (VEGF),Angiopoietin (Ang)-1,Ang-2,angiogenin,angiostatin,basic fibroblast growth factor,and platelet-derived growth factor-BB using protein array technology.The results showed that there were no significant differences in serum concentrations of all 7 angiogenic factors between control group and DM group.Whereas,serum concentrations of V EGF and Ang-2 were significantly increased in DM + UAP group compared with control group [(3 532.10 ± 1 813.72vs 2 444.50 ± 1 152.21) pg/ml,(286.90-± 217.01 vs 171.92 ± 106.63) pg/ml,both P＜0.01].Pearson's correlation analysis revealed that serum concentrations of all 7 angiogenic factors were not related to HbA1C,fasting and 2 h postprandial plasma glucose,triglycerides,high density lipoprotein cholesterol,as well as low density lipoprotein cholesterol.

Objective To detect the changes in serum concentrations of angiogenic factors in patients with unstable angina pectoris.Methods A total of 57 patients with unstable angina pectoris were eligible for study and divided into diabetes group (n =23) and non-diabetes group (n =34).Exclusion criteria included complicated diseases,symptomatic peripheral vascular diseases,renal or liver diseases,cerebral embolism and evidence of ongoing infection.Another 36 age-matched healthy subjects from medical examination center were enrolled as control group.Serum samples were collected,and serum concentrations of vascular endothelial growth factor (VEGF),angiopoietin-1,angiopoietin-2,angiogenin,angiostatin,basic fibroblast growth factor (bFGF) and platelet-derived growth factor-BB (PDGF-BB) were measured by using cytokine array technology.The data were analyzed by independent-samples t test with the SPSS 13.0statistic software package.Results Compared with the control group,the serum concentrations of VEGF and angiopoietin-2 were significantly increased in patients with unstable angina pectoris (both P ＜ 0.01).Whereas,no significant differences in serum concentrations of angiogenin,angiopoietin-1,angiostatin,bFGF,and PDGF-BB were detected between control group and patient groups.There were no significant differences in serum concentrations of above all 7 biomarkers between diabetes group and non-diabetes group.Conclusions Serum concentrations of VEGF and angiopoietin-2 were increased in patients with unstable angina pectoris,and diabetes didn' t affect the increases in serum concentrations of VEGF and angiopoietin2 caused by unstable angina pectoris.

Objective To explore the plasma ghrelin levels in children and adolescents with short stature and the role of ghrelin in growth hormone-releasing hormone-growth hormone(GHRH-GH) axis.Methods One hundred and fifty-seven children(115 male,42 female) with short stature were selected.Fasting plasma sample was extracted from 10 mL vemous blood of the children with short stature.Insulin tolerance test and arginine stimulation test was performed initially to differential diagnosis.And blood samples was divided into 3 ca-tegories:37 cases of complete growth hormone deficiency (CGHD),52 cases of partial growth hormone deficiency(PGHD) and 68 cases of idiopathic short stature(ISS) during these two growth hormone(GH)provocative tests.Controls consisted of age and gender-match 20 health children.Plasma ghrelin levels were measured by radioimmunoassay.Serum GH was detected by chemiluminescence method,and serum insulin-like growth factor-1 (IGF-1) was measured by using enzyme linked immunosorbent assay.Fasting glucose,insulin,testosterone,estra-diol,luteinizing hormone,follicle-stimulating hormone were measured.Statistical analysis were conducted by using SPSS 11.5 software.Results The fasting ghrelin levels of CGHD group were significantly lower than that of ISS group and control group(Pa0.05).The ghrelin levels were positive correlated with the stimulated GH peak(r=0.176 P0.05).Conclusion Ghrelin has an important role on GH secretion and abnormal secretion of ghrelin might be a reason of growth hormone deficiency which due to hypothalamic abnormality.

OBJECTIVE: To investigate the immunoregulation and short-term therapeutic effects of super-selective intra-arterial chemotherapy combined with Fuzheng Kang'ai Granules, a compound Chinese herbal medicine, on patients with late gastric cancer. METHODS: Forty patients with late gastric antrum cancer were randomly divided into study group and control group. Patients in the study group were orally administered Fuzheng Kang'ai Granules 48 hours after the first super-selective left gastric artery chemotherapy with high-dose drugs (EAP regime: VP(16) 100 mg/m(2) + epirubicin 60 mg/m(2) + carboplatin 200 mg/m(2)), while patients in the control group were only administered the same local artery chemotherapy as in the study group. RESULTS: The short-term therapeutic efficacy in the study group and control group were 82.5% and 57.5% respectively (P<0.01). The occurrence rates of side effects in the study group were significantly lower than those in the control group (P<0.01). The Karnofsky score of life quality of the study group was obviously higher than that of the control group after treatment (P<0.05). The half survival time and one year survival rate in the study group and control group were (24.9 +/- 1.36) month, 70% (28/40) and (13.7 +/- 0.72) month, 35% (14/40) respectively. They were significantly improved in the study group compared with the control group (P<0.01). The levels of cytokines interleukin 2 (IL-2), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) after treatment in the study group were significantly higher than those before treatment (P<0.05 or P<0.01), as well as than those after treatment in the control group (P<0.01). On the other hand, the level of immune inhibitory factor soluble interleukin-2 receptor (sIL-2R) after treatment in the study group was lower than that before treatment, as well as than that after treatment in the control group (P<0.01). CONCLUSIONS: The super-selective intra-arterial chemotherapy combined with Fuzheng Kang'ai Granules has good short-term therapeutic efficiency and few side effects for patients with late gastric antrum cancer. It can significantly improve the life quality, extend the survival period, and improve the survival rate in patients with late gastric antrum cancer, which may be due to the up-regulation of the immune regulating factors such as IL-2, TNF-alpha, IFN-gamma and down-regulation of the immune inhibitory factor sIL-2R.

Objective The cytotoxic microbial strains isolated from the deep ocean water and sediments were screened,and the secondary metabolites of bioactive fungus c2b were investi- gated.Methods Active bioactive microbial strains were screened using brine shrimp and chro- nic medulla leucocythemia leukocythemia(K562)cell line.The cytotoxic components of fun- gus c2b were isolated by bioassay-guided fractionation and solvent extraction,silica gel col- umn chromatography and preparative HPLC.Their structures were established by pbysico- chemical properties and spectral analyses.The cytotoxicities of compounds were evaluated by SRB method.Results and Conclusion Twenty-nine strains of fungi were isolated.Among them,seven strains showed cytotoxic activities.Six compounds(1～6)were isolated and i- dentified as N-acetyl histamine(1),chrysogine(2),ergosterol peroxide(3),5,8-epidioxy- 24-methylcholesta-6,22-dien-3?-ol(4)cerevisterol(5)and(4E,8E)-N-[(2'R,3'E)-2'-hy- droxy-3'-hexadecenoyl]-1-O-?-D-glycopyranosyl-9-methyl-4,8-sphingadiene(6),respective- ly.Compound 3 and 4 showed median cytotoxicity.

Objective:To retrospectively analyze the routine clinical laboratory parameters for hepatocellular carcinoma,in an attempt to search for parameters for diagnosis of hepatocellular carcinoma(HCC).Methods:The pre-operation clinical labo- ratory data,such as tumor makers,and serological biochemical indices,hepatitis B virus(HBV)infection markers,and HBV DNA titers,were collected from 828 patients who were pathologically diagnosed as having HCC;then the correlation between these data with tumor size and the pathological grades of HCC was analyzed.Results:It was found that 97.9% of the 828 pa- tients were infected with HBV and 70.9% of them were accompanied by liver fibrosis.We also found that the tumor size was correlated with albumin(ALB),globulin(GLB),A/G,aspartate aminotransferase(AST),ratio of aspartate to alanine amin- otransferase(AST/ALT),gamma-glutamyl transferase(GGT),alkaline phosphatase(ALP),alpha-L-fucosidase(AFU),al- pha-fetoprotein(AFP)and tumor grades;meanwhile,the pathological grades of tumor was correlated to prealbumin(PALB), GGT and tumor size(all P

Objective To study the effect of overexpression of TRPC6 on Ang Ⅱ-induced apoptosis of mouse podocytes in vitro and to explore the possible mechanisms. Methods Mouse TRPC6 cDNA eukaryotie expression vector pEGFP-NI-mTRPC6 was transfected to conditionally immortalized routine podocyte cell line by liposome. The fluorescent microscopy was used to examine the expression of EGFP after 24 hours. The change of TRPC6 protein expression was observed by Western-blot. Podocytes were treated by different concentrations of Ang Ⅱ. The podocyte intracellular calcium concentration was measured with laser-scanning con_focal microscope. The expression of Bax and Bcl-2 mRNA was assessed by RT-PCR and the expression of Bax and Bcl-2 protein was measured by Western-blot. The apoptotic ratio of podocytes was monitored by flow cytometry and Hoechst staining. Results About 35% of the cells expressed EGFP. An up-regulation of protein expression of TRPC6 was detected in podocytes when transfected with pEGFP-N1-mTRPC6 (P<0.01). The overexpression of TRPC6 promoted the Ang Ⅱ-induced influx of extracellular calcium and elevated the expression of Bax but decreased the expression of Bcl-2 (P<0.01, P<0.05). The apoptotic ratio of podocyte was (2.50±0.72)% when treated by low-dose Ang Ⅱ (10-10 mol/L), and it was increased to (4.33±0.45)% when transfected with pEGFP-N1-mTRPC6 (P <0.05 ). Transfection with pEGFP-NI-mTRPC6 increased apoptosis rate from (15.46± 1.40)% to (18.33±0.87)%(P<0.01) by high-dose Ang Ⅱ (10-6 mol/L). Conclusion TRPC6 plays an important role in the Ang Ⅱ-induced apoptosis of podocytes by promoting the influx of extraeellular calcium, which leads to the apoptosis cascade initiation.

Objective To access the effects of aldosterone (ALD) and its receptor antagonist- spironolactone (SPI) on the production of reactive oxygen species (ROS) and apoptosis in podocytes and to explore the possible mechanism involved. Methods Conditionally immortalized mouse podocytes were divided into control group, ALD group, SPI group and SPI +ALD group. The level of ROS production and the expression of nephrin protein were assayed by fluorescent spectrophotometry and indirect immunofluorescence technology. The apoptosis rate of podocytes was monitored by flow cytometry. The expression of bax and bcl-2 mRNA and protein was detected by RT-PCR and Western blotting methods. The anti-oxidant N-acetylcysteine (NAC)was applied to determine whether the effects of ALD on podocytes were mediated by ROS pathway.Results Compared with the control group, ALD significantly increased ROS production in podocytes (P＜0.05). SPI completely abolished the above-mentioned effect of ALD (P＜0.05). ALD induced the down-regulation of the expression of nephrin and the up-regulation of podocytes apoptosis (P＜0.05), which was accompanied with the elevated expression of bax mRNA and protein and the reduced expression of bcl-2 mRNA and protein (P＜0.05). SPI or NAC prevented the above-mentioned changes induced by ALD (P＜0.05). Conclusion ALD increases theexpression of pro-apoptotic factor (bax) but decreases the expression of anti-apoptotic factor (bcl-2)to induce podocytes apoptosis through the mineralocorticoid receptor (MR) possibly via the mechanisms involving ROS pathway.

Objective Complicated aneurysms located in the cisternal segment of the internal carotid artery(ICA-CSA)present unique therapeutic difficulties.This study is to discuss the feasibility of the Willis stent-graft in treating complicated ICA-CSA by comparing its effect with that of coiling therapy.Methods Willis covered stents were employed in 19 complicated ICA-CSAs(group A),while coils were used in 17 complicated ICA-CSAs(group B).Follow-up angiography was performed to investigate aneurysm recurrence,endoleak and parent artery(PA)stenosis.Kaplan-Meier curves were constructed to compare the recurrencefree and PA stenosis-free rate in both groups.Results Total exclusion was immediately achieved in 13 ICACSAs and minor endoleaks presented in 5 cases in group A.Total or near-total occlusion was achieved in 7 ICA-CSAs.subtotal occlusion in 8 and partial occlusion in 2 cases in group B after coiling.Acute thrombosis occurred in 1 patient in either group and re-hemorrhage happened in 1 patient after coiling.Follow-up angiography in group A revealed that 16 ICA-CSAs were completely isolated,with two parent arteries showing mild in-stent stenosis.Eighteen months after the procedure,Kaplan-Meier analysis showed that the recurrence-free rate was 93.3%and 50%,while the stenosis-free rate of parent artery was 87.5%and 100% in group A and in Group B,respectively.In group A and group B the clinical neurological symptoms were fully recovered in 9 and 9,obviously improved in 3 and 5,unchanged in 2 and 2,and aggravated in one and 0 patients,respectively.Conclusion The implantation of Willis stent-graft is a feasible endovascular therapy for complicated ICA-CSAs.When the parent artery is very tortuous or when the risk that a main collateral branch may be wrongly covered and occluded is present,the implantation of Willis covered stent can not be taken as the treatment of first choice.