Objective To investigate the changes in molecule levels of Ras-ERK signal pathway of K562 cells treated with simvastatin in vitro,and to illustrate that simvastatin inducing the changes in molecule levels of Ras- ERK signal pathway is involved in regulation of proliferation and apoptosis of K562 cells. Methods K562 cells,the chronic myelocytic leukemia(CML) cell lines,were cultured and treated with simvastatin in vitro and proliferation activity of K562 cells was detected by MTT. The changes of apoptosis rate and cell cycle of K562 cells were measured by flow cytometry(FCM). The molecular changes of Ras-ERK signal pathway were analyzed by RT- PCR in transcriptional level. Results The proliferation of K562 cells was inhibited by simvastatin,and G_0-G_1 arrested in K562 cells and significant apoptosis rate was observed with FCM. Most molecules of Ras- ERK signal pathway expressed differentially at transctiptional level. Conclusion Simvastatin probablely inhibit proliferation and induces apoptosis of K562 cells,depending on Ras-ERK signal pathway which is involved in cell proliferation and apoptosis.

Objective To analyse the effect of progesterone on peripheral blood corticotropin releasing hormone ( CRH ) and delivery time in women with premature rupture of membranes ( PROM ) .Methods 80 patients who were diagnosed with PROM in Department of Obstetrics and Gynecology, China Medical University were collected.Randomly divided into dexamethasone (DEX) group, dexamethasone plus progesterone (DEX+P) group, progesterone ( P) group and control group, three groups were detected on admission, admission 24 h, 48 h on peripheral white blood cell count, C-reaction protein, CRH level and time of delivery, neonatal weight, and analysis of CRH the level of the correlation and delivery time.Results Compared with the other three groups, the level of CRH in peripheral blood of DEX group were higher (P<0.05);CRH (P<0.05) increased faster;shorter delivery time (P<0.05); the level of CRH was negatively correlated with delivery time (r=-0.832, P<0.05).The results were statistically significant.Conclusion Dexamethasone treatment can make the premature rupture of fetal membranes of peripheral blood CRH levels rise, shorten the delivery time, progesterone can inhibit this process.

Objective To study the findings of measles associated with pneumonia and to improve the diagnostic level of this disease.Methods The chest X-ray of 280 cases with pneumonia among 310 children with measles from January 2005 to August 2006 were collected and analyzed retrospectively.Results The most common findings of chest X-ray examinations included blurred and increased lung markings(in all cases),pulmonary air-sacs luminance increase and emphysema-like change in 206 cases(73.6%),spotted shadows and patches in the bilateral middle and lower field in 91 cases(32.5%),enlarged and blurred pulmonary hila in 115 cases(41.1%),interstitial appearance in 98 cases(35%),serious pneumonia and ARDS in 4 cases(1.4%).Conclusion It is valuable in the diagnosis and differential diagnosis of measles accompanying pneumonia in children to analyze the chest X-ray combining with clinical situation.

Objective To study the expression and clinical significance of Notch intracellular domain (NICD) in cervical cancer and the effects of N-[N-(3,5-difluorophenyl)acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis of cervical cancer cell lines. Methods Western blot was used to detect the expression of NICD in the tissues of 40 cervical cancers and 21 normal cervix and its relationship with clinical features of cervical cancer was also analyzed. Proliferation of SiHa and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in SiHa and HeLa cells incubated with DAPT was detected by western blot. Results The expression level of NICD in cervical cancers was significantly higher than that of normal cervical tissues (1.237±0.353 vs 0.938±0.105, P<0.05). The NICD expression was higher in cervical cancers with high grade,lymph node involvement and parametrial invasion than that with low-middle grade (1.496±0.540 vs 1.150±0.216), without lymph node involvement (1.419±0.532 vs 1.159±0.210) and no parametrial invasion (1.718±0.710 vs 1.183±0.258), respectively (all P<0.05). The expression of NICD in cervical adenocarcinoma was higher than that of squamous cell cancer (1.463±0.395 vs 1.162±0.187, P<0.05). After SiHa and HeLa cells were incubated with DAPT, NICD expression was significantly lower than that in control (P<0.05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells was depended on its concentrations and times. Conclusions NICD may play a key role in the occurrence and progress of cervical cancer. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells may be due to decreased the formation of NICD.

Angiotensin II (ANGII) plays an important role in the pathogenesis of atherosclerosis by inducing proliferation of vascular smooth muscle cells (VSMCs). In our study, we observed the effects of valsartan on proliferation of cultured VSMCs treated with or without ANGII by cell counting and methyl thiazolyl tetrazolium (MTT) assay, and detected the expression of mitofusin 2 (Mfn2), a newly discovered cell proliferation inhibitor and a related cell proliferation signaling pathway protein by Western blotting. ANGII at a concentration of 10(-6) mol/L significantly stimulated VSMCs proliferation, down-regulated the expression of Mfn2 and up-regulated the expression of Raf and ERK1/2. Valsartan inhibited such effects of ANGII at concentrations of 10(-5) and 10(-6) mol/L, but not at 10(-7) mol/L. Valsartan had no significant effect on the proliferation of untreated VSMCs. These results suggest that valsartan inhibits ANGII-induced proliferation of VSMCs in vitro via Mfn2-Ras-Raf-ERK/MAPK signaling pathway.

Objective In this study ,to explore the relationship between the renal blood flow PI ,and the AKI was caused by CPB .Methods 14 cases with heart disease were accepted .The renal aorta and renal segmental artery PI of all cases were monitored by the CDFI at the preoperative and postoperative 1 h ,2 h ,4 h ,8 h ,16 h ,24 h .The renal blood urea nitrogen (Urea) ,uric acid (UA) ,creatinine (Crea) ,were detected at the same time .All datas for statistical analysis .Results The renal aorta PI was higher at the postoperative 1 h ,2 h ,4 h ,8 h ,16 h than that at the preoperative .The renal segmental artery PI was higher at the postoperative 1 h ,2 h ,4 h ,16 h than that at the preoperative .The renal blood flow PI was positively correlated with Urea ,UA and Crea .Conclu-sion The renal aorta ,renal segmental artery PI was positively correlated with Urea ,UA and Crea after CPB .The PI may be seen as an evaluation index to assess AKI after CPB .

AIM: To probe into tau hyperphosphorylation at PHF- 1 sites induced by glycogen synthase kinase - 3β(GSK- 3β) in vivo. METHODS: Twenty - one rats were randomly allocated to three groups as follows: GSK - 3β transfection group, vector group and control group; 0.1 μg/3μL GSK- 3β- HA plasmid or vector was injected bilaterally into cerebrum of the rats respectively, rats without injection were controls. Western blotting and immunohistochemical staining of cortex were carried out to detect the expression of GSK- 3β- HA plasmid and tau phosphorylation using phosphorylation- dependent tau antibody PHF- 1. RESULTS: After transfection with GSK- 3β- HA for 48 h, GSK - 3β - HA was expressed in GSK- 3β transfection group; and hyperphosphorylated tau at PHF- 1 sites accumulated in neurons in the transfected areas. The hyperphosphorylated tau colocalized largely with GSK- 3β expressed by the transfected GSK- 3β plasmid. CONCLUSIONS: Transfection with GSK- 3βin vivo can induce tau hyperphosphorylation involving the pathogenesis of neurodegenerative disorders. These data further prove that GSK- 3β is a key kinase to induce tau hyperphosphorylation and may be a therapeutic target for tauopathy- related neurodegenerative diseases.

Objective To determine the effect of community family visit on patients with metabolic syndrome.Methods According to the diagnostic criteria set up by the Diabetes Society of the Chinese Medical Association 220 patients with metabolic syndrome were equally divided into two groups:family visit group and control group.The family visit group was followed up by full-time medical staff regularly,while not interfering with the control group.After one year before and after the intervention,the relevant indicators were compared between the two groups.Results The levels of SBP,DBP,TG,TC,LDL-ch,24-hour urine protein were lowered markedly by intervention (P＜0.05).HDL-ch increased compared to the previous (t= 7.921,P＜0.05),but body mass index were not significantly changed.Before and after the intervention the levels of SBP,DBP,TG were ideal This was followed by significant improvement of fasting plasma glucose (FPG),two hours after meal blood glucose (2 hPG),TC,HDL set standards,24-hour urine protein body mass index.Compared with the control group,the family visit group showed siguificant improvement of related indicators except body mass index and TC.Condusion Intervention by family visit is effective in improving the vales of metabolic syndrome.

Objective To investigate the effect of tMfn2 gene on inhibiting the proliferation of vascular smooth muscle cells (VSMCs) and related mechanism. Method VSMCs were transfected with adenovirus vector encoding tMfn2 or Mfn2 (Adv-tMfn2, Adv-Mfn2). The abundance of tMfn2 protein and Mfn2 protein were deter-mined by Western blot analysis using Mfn2 N-term antibody. The effect of tMfn2 on the proliferation of VSMCs was explored by cell counting and MTT assay. The cell cycle was analyzed using flow cytometry. Western blot were used to detect the expression of p-ERK1/2 and p-Raf-1. Results The results of cell counting and MTT both indi-cated that tMfn2 gene displayed more remarkable effect on inhibiting the proliferation of VSMCs than Mfn2 (P <0.01). Flow cytometry showed that most of the cells infected with Adv-tMfn2 or Adv-Mfn2 were blocked in the stage of G0/G1 and few entered into the S phase. Western blot indicated that overexpression of tMfn2 gene resulted in downregulation of phosphorylated ERK1/2 and Raf-1 protein (P < 0.01). These results demonstrated tMfn2 had stronger effect than Mfn2 (P < 0.01). Conclusions tMfn2 gene is superior to Mfn2 gene in attenuating the proliferation of VSMCs via the Ras-Raf-ERK1/2 signaling pathway.

Objective: To prepare adipose-derived stem cells (ADSCs) and chitosan chloride (CSCl) gel complex to study the biocompatibility and the feasibility of repairing the wounds of deep partial thickness scald in rats. Methods: ADSCs were prepared by enzymogen digestion and differential adherence method from the subcutaneous adipose tissue of SPF grade 6-week-old male Sprague Dawley (SD) rats. Temperature sensitive CSCl gel was prepared by mixing CSCl, β glycerol phosphate, and hydroxyethyl cellulose in 8∶2∶2.5 ratio. The proliferation of ADSCs was measured by cell counting kit 8 (CCK-8) assay and the survival of ADSCs was detected by the Live/Dead flurescent staining in vitro. A deep partial thickness burn animal model was made on the back of 72 SPF grade 6-week-old male SD rats by boiled water contact method and randomly divided into 3 groups ( n=24). Group A was blank control group, group B was CSCl hydrogel group, group C was ADSCs/CSCl gel group. The wound closure rate at 3, 7, 14, 21 days was observed after operation. The number of inflammatory cells at 7 days and epidermal thickness at 21 days were observed by HE staining after operation. The angiogenesis at 7 days was evaluated by immunohistochemistry staining with CD31 expression. Results: CSCl had a temperature sensitivity, at 4℃, the temperature-responsive hydrogel was liquid and became solid at 37℃. The CCK-8 assay and Live/Dead flurescent staining confirmed that ADSCs could grow and proliferate in the ADSCs/CSCl hydrogel complex. General observation showed the wound closure ratio in group C was superior to groups A and B after operation ( P<0.05). HE staining showed that at 7 days after operation, the wound healing of the three groups entered fibrous proliferation stage. Collagen deposition and inflammatory cell infiltration were observed in the dermis of each group. The proportion of inflammatory cells in group C was significantly lower than that in groups A and B, and in group B than in group A ( P<0.01). At 21 days after operation, the fibrous connective tissues of neoepithelium and dermis in groups B and C were arranged neatly, and fibroblasts and neocapillaries could be seen. In group A, neoepidermis could also be seen, but the fibrous connective tissues in dermis were arranged disorderly and sporadic capillaries could be seen. The thickness of neonatal epidermis in group C was significantly larger than that in groups A and B, and in group B than in group A ( P<0.01). CD31 immunohistochemistry staining showed that the neovascularization could be seen in all groups. The number of neovascularization in group C was significantly higher than that in groups A and B, and in group B than in group A ( P<0.05). Conclusion: The ADSCs/CSCl hydrogel complex has a good biocompatibility and possessed positive effects on promoting the deep partial thickness scald wound repairing in rats.