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Objective To detect phosphorylated-extracellular signal-regulated kinase (p-ERK1/2) protein expression in epithelial ovarian cancer and cell lines,and to examine the effects of mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor AZD6244 on cell proliferation,apoptosis as well as cell cycle of ovarian cancer cells.To explore the function and significance of MAPK/extracellular signal-regulated kinase (ERK) signaling pathway in the development of ovarian cancer.Methods (1) A total of 104 cases of patients with ovarian cancer who accepted the treatment of gynecological surgery and being confirmed by pathological examination in First Affiliated Hospital,Dalian Medical University from January 2004 to December 2013 were selected.The expressions of p-ERK1/2 protein were detected by immunohistochemistry in ovarian cancer specimens,and the relationship between the expressions of p-ERK1/2 and the clinical features of patients was analyzed.(2) p-ERK1/2 and other related proteins were determined by western blot in various ovarian cancer cells,including SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3 treated with or without MEK inhibitor.The cellular proliferation,apoptosis and cell cycle of ovarian cancer cells after treatment with MEK inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry,respectively.Results (1) The immunohistochemical method showed that p-ERK1/2 between low grade serous carcinoma and clear cell carcinoma were not significantly higher expressed (P＞0.05).However,a lower level of the p-ERK1/2 expression were observed among high grade serous carcinoma,mucinous carcinoma and endometrioid carcinoma (all P＜0.05).There was no significant correlation between the protein expression of p-ERK1/2 and patients' age,pathological stage of surgery,and preoperative serum CA125 level (P＞0.05).(2) Western blot showed that the protein p-ERK1/2 was widely expressed in various ovarian cancer cell lines such as SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3.After treatment with AZD6244 (5,10 μmol/L),the level of p-ERK 1/2 in OVCAR5 and OVCAR8 decreased significantly in dose-dependent manner.Additionally,we found a reduction of the expression level of cyclin D 1,caspase-3 and appeared cleaved poly adenosine diphosphate ribose polymerase (PARP) in OVCAR5 and OVCAR8,compared with control groups.MTT assays showed that OVCAR5,OVCAR8 and A2780S were differently inhibited in the dose-dependent manner after being treated with different concentrations of AZD6244 (0,2.5,5,10,25,50 and 100 μmol/L,all P＜0.05).Further tested by flow cytometry,the results showed that AZD6244 (5,10 μmol/L) was able to induce the apoptosis of OVCAR5,OVCAR8 and A2780S,as well as G0/G1 phase arrest,both in a dose-dependent manner (P＜0.05).Conclusions As the main active and functional unit of MAPK/ERK signaling pathway,p-ERK1/2 protein is expressed in both the tissues and various ovarian cancer cell lines.AZD6244 could down-regulated the expression of p-ERK1/2 in ovarian cancer cells,accompanied by the decreased proliferation and increased cell apoptosis of ovarian cancer cells.In conclusion,MAPK/ERK signaling pathway might play a role in the development and progression of ovarian cancer,and may be provide a novel option for molecular targeted therapies of the disease.
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Objective To observe the protective effects of Cordyceps Extract (Cordyceps sinensis, CS) on ischemia reperfusion injury of transplanted kidney in renal allograft rejection, and to investigate the inhibition of immune regulation mechanism.Methods 100 rats were randomly divided into a blank control group, an ischemia reperfusion group, a cyclosporine A group, a Cordyceps high dose group (CSA), and a Cordyceps small dose group (CSB). Except the blank control group, all the other groups were underwent same race variant body left renal transplantation. 2 hours after the transplantation, cyclosporin A 2.0 mg/(kg?d) was injected to the cyclosporine group, CSA group, and CSB group. 1% CS of 5 and 10 ml/kg by gavage was respectively given to CSA and CSB groups. 10ml/kg sodium chloride was injected to the bland control group and the ischemia reperfusion group. After 14 days (d) of administration, the content of blood urea nitrogen (BUN), Cystatin C (Cys C), creatinine (Cr), total superoxide dismutase (T-SOD), serum malondialdehyde(MDA), lactate dehydrogenase (LDH), serum total protein (TP), albumin (ALB) were detected. Results Compared with the cyclosporin A group, the content of BUN(8.71 ± 0.87mmol/L, 9.78 ± 1.01mmol/Lvs.12.59 ± 1.54mmol/L), Cys C(1.36 ± 0.09 mg/L, 1.40 ± 0.11mg/L vs.1.94 ± 0.12 mg/L), Cr(117.79 ± 15.35μmol/L, 121.28 ± 16.01μmol/L vs.133.76 ± 17.07 μmol/L), LDH(157.32 ± 20.93U/L, 154.35 ± 19.39U/L vs.187.78 ± 25.18 U/L), MDA(122.23 ± 19.41nmol/ml, 125.29 ± 25.97nmol/ml vs.184.23 ± 26.74 nmol/ml) were decreased in the CSA group and CSB group, T-SOD(80.35 ± 7.71U/ml, 78.30 ± 7.79U/ml vs.56.43 ± 7.25 U/ml) was increased(P<0.05).Conclusion CS can significantly inhibit lipid peroxidation, renal ischemia reperfusion caused a decrease in MDA and LDH production, increase the activity of T-SOD and TP, ALB, reduce the damage of kidney function, have a dual immunomodulatory effects obvious, with cyclosporin A anti rejection has synergistic effects, while increasing the body immunity, to allograft renal transplantation has protective effect of renal ischemia / reperfusion injury in rats.
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Objective To estimate heart function among patients with diabetic foot ulcer,and to investigate the characteristics of heart failure(HF) before and after treatment of ulcer. Methods Items associated with diabetes and some physiological and biochemical indicators were observed in patients with diabetic foot ulcer(162 cases) and with high risk factors of ulcer(75 cases). Heart function was evaluated at patients′ admission, during ulcer related treatment, and prior to discharge. Left ventricular ejection fraction and other cardiac assessment were measured with ultrasonic scan.Results During hospitalization, 23.6%(56/237) patients underwent HF with normal left ventricular ejection fraction, and it was 27.2%(44/162) in patients with foot ulcers. The prevalence of HF was 8.9%(21/237) in all patients studied on admission, and that was 10.5%(17/162) in patients with foot ulcer, more than that without foot ulcer(P＜0.01). More patients with HF were found, being 14.8%(35/237) during 2-7 days after ulcer related treatment initiated, and peak was on the 4th day. There was statistical difference among different Wagner grades(P＜0.05) about the morbidity rate of HF. All patients with HF were improved and tolerant to ulcer related treatment. Conclusion The prevalence of HF with normal left ventricular ejection fraction was relatively high in patients with diabetic foot ulcer, especially after ulcer treatment.
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Objective:To study the effect of midnight-noon ebb-flow acupoint selection on rheumatoid arthritis. Methods:120 cases of patients with rheumatoid arthritis were randomly divided into three groups:midnight-noon ebb-flow acupoint selection group,acupuncture by differentiation of symptoms and signs group,integrative group of midnight-noon ebb-flow acupoint selection and acupuncture by differentiation of symptoms and signs group. Results:In the midday-midnight flowing of qi-blood acupuncture group of 40 cases,11 cases were improved obviously,11cases were improved,9cases were improved slightly,9 cases were ineffective,the total effective rate was 77.5%. In the acupuncture by differentiation of symptoms and signs group of 40 patients,10 cases were improved significantly,11 cases were improved,11 cases were improved moderate,8 cases were ineffective,the total effective rate was 80.0%. In integrated group of midnight-noon ebb-flow acupoint selection and acupuncture by differentiation of symptoms and signs group of 40 patients,15 cases were improved significantly,13 cases were improved,10 cases were improved moderate,2 cases were ineffective,the total effective rate was 95.0%. Curative effect of the integrative group was better than that of other two groups(P