Literature review to build optional indexes pool,Delphi method to screen indexes,fuzzy model used to calculate the weight,for building such a system.An index system centering on public wellbeing is thus constructed,comprising three level 1 indicators,six level 2 indicators,40 level-3 indicators.The system can satisfactorily reflect the vision of county-level public hospitals reform,and showcase outcomes of such a reform.

Objective To explore the clinical application of pereutaneous transluminal angioplasty(PTA) and endovascular bracket to treat lower extremity arteriosclorotic occlusion.Methods The clinical data of 36 patients(41 affected limbs)with lower extremity atherosclerotic occlusion who were treated with PTA and bracket implantation from Jan 2008 to Dee 2008 were summarized.Results The initial successful rate of PTA wag 95.1% (39/41).The clinical symptoms were considerably improved in 37 affected limbs,representing of pain disappearing,skin temperature increasing and the healing of refractory ulcer.The index of ankle to brachial significantly increased from 0.54±0.11 to 0.79±0.15(before v.s.after therapy).However,no improvement was observed in 3 affected limbs,and one affected limb Wag re-operated by the amputation.In the following 3 to 15 months.three superficial femorsI arteries were re-obstructed at the 5th,6th,12th month,respectively.One arteria tibialis pesterior was re-obstructed at the 8th month.The cumulative cure rate was 89.7%(35/39).Conclusions PTA is effective in treating atherosclerotic occlusive diseases.The endovascular bracket can increase the cumulative cure rate.PTA and endovascular bracket are safe and effective in treating lower extremity arteriosclerotie occlusion.

BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.

Background and purpose:The prognosis in patients with early breast cancer(EBC) was poorly defined by clinical and histopathological features.There were more than 200 genes that might be correlated with development,progression,recurrence and prognosis of breast cancer in the published literature.This study evaluated the prognosis of patients with EBC at molecule level according to the protein expressions of 16 selecting genes(HER2,ER,PR,BCL2,Ki-67,BMYB,Cyclin B1,STK15,MMP11,BAG1,NM23,PTEN,P53,P27,VEGF,PCN A).Methods:The immunohistochemical method was used to detect the protein expressions of these genes in 76 patients with EBC and the statistical analysis was performed by COX proportional hazards model.Results:COX proportional hazard model revealed that BCL2,VEGF and STK15 were independent prognostic factors for patients with EBC.A formula of PI was set up according the three genes.The predictive outcomes with PI were compared with the actual follow-up outcomes in 48 patients with EBC.As a result,the predictive accuracy of good and bad prognosis was 86.67% and 91.67%,respectively.Conclusion:A formula of PI derived from protein expressions of the 16 genes correlated with breast cancer was ultimately set up and might be used to predict the outcomes of the patients with EBC.

OBJECTIVE: To observe the effects of Qingdan Capsule (QDC) and Yanggan Lidan Granule (YGLDG), two kinds of compound traditional Chinese herbal medicines, on biochemical parameters in guinea-pigs with pigment gallstones. METHODS: An animal model of pigment gallstones was established in male guinea-pigs by hypodermic injection of lincomycin. The guinea-pigs were randomly divided into blank control group, untreated group, QDC group and YGLDG group. There were 8 guinea-pigs in each group. After ten-day treatment, animals were sacrificed and sampled to calculate the rate of stone formation, total bilirubin (TB), unconjugated bilirubin (UCB) and Ca2+ density in bile of the four groups. RESULTS: In comparison with the untreated group, the rates of stone formation in the QDC and YGLDG groups were significantly decreased (P<0.01). TBIL, UCB and Ca2+ content of bile in both QDC and YGLD groups was also significantly decreased (P<0.05, P<0.01). CONCLUSION: QDC and YGLD have good effects on biochemical changes of animal model of pigment gallstone in reversing the lithogenesity of bile by reducing the content of TB, UCB and Ca2+, hence resulting in clinical treatment and prevention of pigment gallstone disease.

Objective To investigate the effect of proliferation,apoptosis and cell cycle of 3-MA on rat endothelial progenitor cells. Methods Bone marrow-derived mononuclear cells were isolated from rat bone marrow by ficoll. There were five groups. The control group and four 3-MA concentration groups: 1. 25 mmol/L,2. 5 mmol/L,5 mmol/L, 10 mmol/L. MTT was used to measure the proliferation of endothelial progenitor cells. Flow cytometry ( FCM) was used to detect cell apoptosis and cell cycle. Results (1)5 mmol/L 3-MA promotes proliferation of endothelial progenitor cells, while 10 mmol/L 3-MA inhibits the proliferation of endothelial progenitor cells (P ＜ 0. 05). (2) 10 mmol/L 3-MA promotes apoptosis of endothelial progenitor cells, compared with the control, the difference was significant ( P ＜ 0. 05 ). (3) 3-MA at the concentration of 5 mmol/L reduces cells at G0/G1 phase and increases S and G2/M phase cells; 10 mmol/L 3-MA induces endothelial progenitor cells blockade at S phase, G2/M phase cells decreased, compared with the control, the difference was significant (P ＜ 0. 05). Conclusions 5 mmol/L 3-MA promotes the proliferation of endothelial progenitor cells. 10 mmol/L 3-MA inhibits the proliferation and promotes apoptosis of endothelial progenitor cells.

With IL-6R as target, a new compound 2460A was identified from fungus using HTS screening model. The taxonomics of the produced strain was confirmed to be Trichoderma hazianum rifai after sequencing analysis of rDNA-ITS (internal transcribed spacer). Results showed that this compound has a binding activity on IL-6R competed with IL-6, thus it is a new ligand of IL-6R originating from microbe. With MTT assay, the anti-tumor activities of 2460A were demonstrated on CM126 and HT-29 cell lines separately, the IC50 are 2.17 x 10(-5) mol x L(-1) and 1.8 x 10(-5) mol x L(-1) respectively. The compound affected lightly the HT-29 cell cycle at S phase. Studies for the anti-tumor activity of 2460A in vivo are in progress in our lab.

Objective To explore the effect of hypoxia inducible factor-1α silencing by siRNA on proliferation, apoptosis and necrosis of human renal proximal epithelial cell ( HK-2 )under hypoxia. Methods CoCl2 was used to mimic hypoxia, and siRNA technique was used to silence HIF-1α. HK-2 cells were divided into five groups, including normal culture group,hypoxia culture group, transfection reagent group, negative control group and HIF-1α siRNA group.MTT assay was used to detect the growth inhibition ratio of cells. TUNEL and caspase-3 quantity was used to detect apoptosis. LDH activity in supernatant was detected to evaluate cell necrosis.Real-time PCR and Western blotting were used to detect mRNA and protein of HIF-1α, HIF-2α,glucose transporter 1(Glut-1) and vascular endothelial growth factor (VEGF). Results (1) The transfection efficiency of siRNA was 95%-100%. Under normoxia, the efficiency of siRNA silencing HIF-1α gene reached to 70%, while under hypoxia, it was 97%. (2) The growth inhibition ratio of cells in HIF-1α siRNA group was significantly higher than that of other groups including hypoxia culture group, transfection reagent group and negative control group (all P＜0.05). No significant difference was found in apoptotic ratio of the other four groups except normal culture group (P＞0.05). The LDH level in HIF-1α siRNA group was significantly higher than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). (3) The expression of HIF1α, Glut-1, VEGF mRNA and protein in HIF-1α siRNA group was significantly lower than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). No significant difference was observed on the level of HIF-2α mRNA and protein in the other four groups except normal culture group (P＞0.05). Conclusion HIF-1α gene silencing can inhibit the mRNA and protein expression of Glut-1 and VEGF, and can aggravate growth inhibition and necrosis of HK-2 cells under hypoxia.

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

OBJECTIVE: To investigate the effects of Qufeng Tongluo Recipe (QFTLR), a compound of traditional Chinese herbal medicine for dispelling wind and removing obstruction in the meridians, on cell proliferation and expressions of transforming growth factor-beta1 (TGF-beta1) and interleukin-6 (IL-6) mRNAs induced by lippolysaccharide in glomerular mesangial cells from rats. METHODS: The method of serum pharmacology was used. A total of 32 rats were divided into normal control group, untreated group, QFTLR group and positive control group which also was named Monopril (fosinopril sodium) group. Mesangial proliferative glomerulonephritis was induced by injection of antithymocyte serum in the rats except for the normal control group. Sera of the rats were obtained after corresponding interventions. Lipopolysaccharide-induced proliferation of rat mesangial cells (MCs) cultured in the respective serum-containing media was detected by the method of methyl thiazolyl tetrazolium (MTT) assay, and the expressions of TGF-beta1 and IL-6 mRNAs were analyzed by the method of reverse transcription polymerase chain reaction. RESULTS: Compared with the untreated group, QFTLR showed remarkable inhibitory function on the proliferation of the mesangial cells (P<0.05). The expressions of TGF-beta1 mRNA in mesangial cells were increased in the untreated group, QFTLR group and Monopril group when compared with the normal control group (P<0.01), but the TGF-beta1 mRNA expressions in QFTLR group and in Monopril group were lower than that in the untreated group. The IL-6 mRNA expression could be increased by the LPS stimulation, and it was significantly higher in the other three groups than that in the normal control group, including the untreated group, the Monopril group and the QFTLR group (P<0.01). Compared with the untreated group, the expression of IL-6 mRNA was decreased by QFTLR and Monopril (P<0.01). QFTLR was better than Monopril in inhibiting the proliferation of the mesangial cells and decreasing the expressions of TGF-beta1 and IL-6 mRNAs (P<0.05). CONCLUSION: QFTLR has great inhibitory effect on mesangial cell proliferation and expressions of TGF-beta1 and IL-6 mRNAs, which may be one of its mechanisms in postponing glomerular sclerosis.