Objective:To determine the effects of silencing glucose regulated protein ( GRP94 ) on the proliferation and apoptosis of breast carcinoma MCF7 cells. Methods:Chemically synthesized siRNA targeting GRP94 gene and transfected into MCF7 cells used by Liopfectamine RNAIMAX. The mRNA and protein expression levels of GRP94,cyclinD1,Bax and Bcl-2 were detected by Real-time PCR and Western blot. CCK8 assay was used to detect the effect of specific GRP94 siRNA on cell proliferation and the effect on cell cycle and apoptosis were analyzed by flow cytometry and Hoechst 33258 staining. Results:Compared with the siRNA-NC cells, the expression of GRP94 was significantly down-regulated in MCF7 cells. Knockdown of GRP94 in MCF7 cells decreased cell proliferation and promoted cell apoptosis. The expression of cyclinD1and Bcl-2levels were significantly reduced, and Bax level was increased in siRNA-GRP94 MCF7 cells. Conclusion: The siRNA-mediated GRP94 silence significantly inhibits MCF7 cell proliferation,promoted cell apoptosis by down-regulating cyclin D1,Bcl-2 expression and up-regulating the Bax expression in MCF7 cells.

Animals ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Liver Cirrhosis ; drug therapy ; Phytotherapy ; Pulmonary Fibrosis ; drug therapy ; Rheum ; Salvia miltiorrhiza ; chemistry

ObjectiveTo investigate the effects of morphine preconditioning on myocardial ischemiareperfusion (I/R) injury and the expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2)in rats with chronic heart failure.MethodsForty-eight healthy male SD rats weighing 220-250 g were randomly divided into 6 groups ( n =8 each):control group (group C),sham operation group (group S),I/R group and preconditioning with low,median and high doses of morphine groups (groups MP1-3 ).Chronic heart failure was induced by iv edriamycin 2.0 mg/kg once a week for 6 weeks in groups S,I/R and MP1-3.Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were measured using ultrasound,and left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were calculated at the end of 14th day after the end of adriamycin administration.Blood samples from the carotid artery were collected after ultrasonography for determination of the plasma brain natriuretic peptide (BNP) concentration.Myocardial I/R was induced by 30 min occlusion of left anterior descending branch of coronary artery followed by 120 min reperfusion at 2 day after ultrasonography in groups I/R and MP1-3.In groups MP1-3,iv morphine 0.015,0.030 and 0.050 mg/kg were repeated 3 times at 5 min interval at 30 min before ischemia respectively,while normal saline 5 ml/kg was given in group I/R.The animals were sacrificed at the end of reperfusion in groups S,I/R and MP1-3,and the hearts were removed to measure the area at risk (AAR),infarct size (IS),and IS/AAR ratio was calculated.The p-ERK1/2 expression in myocardium was assessed by Western blot.ResultsThe LVESD and plasma BNP concentration were significantly higher,while the LVEF and LVFS lower in the other 5 groups than in group C (P ＜0.01).No myocardial infarction was found in group S.The p-ERK1/2 expression was significantly lower in groups I/R and MP1 than in group S (P ＜ 0.05).IS and IS/AAR ratio were significantly lower,and p-ERK1/2expression was significantly higher in groups MP2.3 than in group I/R ( P ＜ 0.05).There were no significant differences in IS,IS/AAR ratio and p-ERK1/2 expression between groups MP1 and I/R (P ＞ 0.05).IS and IS/AAR ratio were decreased gradually,and the p-ERK1/2 expression was up-regulated gradually in groups MP1-3 ( P ＜0.05).ConclusionMorphine preconditioning can confer cardioprotection against myocardial I/R in a dose-dependent manner in rats with chronic heart failure.Up-regulation of p-ERK1/2 expression is involved in the underlying mechamism.

Objective:To investigate the effects of ALEX1 overexpression on cell proliferation and apoptosis of human breast cancer cell line MCF-7.Methods: MCF-7 cells were infected recombinant lentivirus LV5-ALEX1 and the negative control lentivirus LV5-NC,respectively.After 72 h, the expression of ALEX1 was detected by Real-Time PCR and Western blot.CCK8 assay were performed to observe the proliferation ability after 24 h, 48 h, 72 h, 96 h.The effect of overexpression ALEX1 on apoptosis was determined by flow cytometry.The level of Bax,BCL-2 and active caspase3 was detected by Western blot.Results:The mRNA level of ALEX1 markedly increased after 72 h(165.81±12.14 vs 52.29±2.32,P<0.01).In CCK8 assay,the results revealed that the cell pro-liferation was inhibited compared with control group in 48 h,72 h,96 h( P<0.05).The results revealed that overexpression of ALEX1 enhanced MCF-7 apoptosis(20.55%±2.50 % vs 3.60%±1.614%,P<0.05).The results by Western blot showed that the protein levels of Bax and active caspase were increased in LV5-ALEX1 group compared with LV5-NC group.However,the protein levels of BCL-2 was decreased in LV5-ALEX1 group compared with LV5-NC group.Conclusion:Overexpression of ALEX1 significantly reduced MCF-7 cancer cells proliferation and induced MCF-7 cells apoptosis.

Objective To investigate the relationship between the expressions of angiotensin converting enzyme 2(ACE2)/ACE mRNA in myocardium-nephors and the blood pressure in spontaneously hypertensive rats (SHR).Method A total of 18 male of 12 weeks old SHRs and 18 male of 12 weeks old Wistar Kyoto(WKY)rats were got for the experimental study.Nine SHRs and 9 WKY rats were randomly taken out as controls and sacrificed for detecting the mRNA expressions and protein of ACE2 and ACE.Other halves of rats of both groups were fed for 12 weeks and then sacrificed for detecting of the same biomarkers as in controls.The mRNA expressions of ACE2 and ACE were determined by real-time quantitative RT-PCR.The protein level of ACE2 and ACE were determined by immunohistochemistry.Results Systolic blood pressure(SBP)in SHRs of 12 and 24 weeks old increased compared with that in WKY(all P＜0.01).SBP in SHRs of 24 weeks old increased compared with that in SHRs of 12 weeks old(P＜0.01).The cardiac and renal expressions of ACE2 mRNA in SHRs of 12 and 24 weeks old were all significantly lower than those in WKY rats(P＜0.01).The expressions of ACE mRNA in SHRs of 12 and 24 weeks old were all significantly higher than those in WKY rats(P＜0.05,P＜0.01).The cardiac and renal expression of ACE2 mRNA in SHRs of 24 weeks old was significantly lower than that in SHRs of 12 weeks(P＜0.01).The myocardial and renal expression of ACE mRNA in SHRs of 24 weeks old was significantly higher than that in 12 SHRs of weeks old(P＜0.01).The positive reactions of ACE2 protein found in kidney displayed in brown colur.The ACE2 proetin appeared in uniform distribution of brown granules in WKY rats and comparatively scanty distribution of brown granule in SHRs.Conclusions There are correaltions between the expressions of ACE2/ACE mKNA as well as protein and blood pressure.

Objective To evaluate the effect of oxycodone preconditioning on liver injury induced by intestinal ischemia-reperfusion (I/R) in rats and the role of different opioid receptors.Methods Fiftyfour adult male Sprague-Dawley rats, weighing 200-300 g, were randomly divided into 9 groups (n =6 each) using a random number table: sham operation group (group S), group I/R, oxycodone preconditioning group (group OP) , μ receptor antagonist CTOP group (group CTOP) , δ receptor antagonist naltrindole group (group NTD), κ receptor antagonist nor-binaltorphimne group (group BNI), CTOP + oxycodo ne preconditioning group (group CTOP+OP) , naltrindole + oxycodone preconditioning group (group NTD+ OP) , and nor-binaltorphimne + oxycodone preconditioning group (BNI+OP).The model of intestinal I/R was established by occlusion of the superior mesenteric artery for 45 min followed by 2 h reperfusion in anesthetized rats.The superior mesenteric artery was only exposed, but not occluded in group S.In OP,COTP+OP, NTD+OP and BNI+OP groups, oxycodone 0.5 mg/kg was injected intravenously at 10 min prior to ischemia.COTP 1 mg/kg and naltrindole 5 mg/kg were injected intravenously at 20 min prior to ischemia in COTP+OP and NTD+OP groups, respectively.Nor-binaltorphimne 5 mg/kg was injected intravenously at 25 min prior to ischemia in group BNI+OP.In CTOP and NTD groups, the corresponding doses of CTOP and naltrindole were injected intravenously at 10 min prior to ischemia.In group BNI, the corresponding dose of nor-binaltorphimne was injected intravenously at 15 min prior to ischemia.The rats were sacrificed at 2 h of reperfusion, and left hepatic lobes were removed for microscopic examination and for detection of apoptosis in liver cells (using TUNEL).The apoptosis index (AI) was calculated.Results Compared with group S, the AI was significantly increased in the other groups (P＜0.05).Compared with group I/R, the AI was significantly decreased (P＜0.05) , and the pathological changes of livers were reduced in OP, COTP+OP, NTD+OP and BNI+OP groups, and no significant change was found in AI and pathological changes of livers in CTOP, NTD and BNI groups (P＞0.05).Compared with group OP, the AI was significantly increased (P＜0.05), and the pathological changes of livers were aggravated in COTP+ OP, NTD+OP and BNI+OP groups.There was no significant difference in AI and pathological changes of livers among groups COTP+OP, NTD+OP and BNI+OP (P＞0.05).Conclusion Oxycodone preconditioning can mitigate liver injury induced by intestinal I/R in rats, and μ, δ and κ receptors mediate the role with comparable effects.

Objective To investigate the risk factors of hepatitis B virus (HBV) reactivation after microwave ablation (MWA) and its prevention.Methods 72 patients who met the inclusion criteria were enrolled into the study.30 patients were in the control group and 42 patients in the prophylactic antivirus group.Results 8.3％ (6/72) patients developed HBV reactivation.A high HBV DNA load and no prophylactic antivirus therapy were independent risk factors of viral reactivation.Conclusion Prophylactic antivirus therapy can prevent HBV reactivation.

The purity of 4,4′-dimethoxy-5,6,5′,6′-bis (methylenedioxy)-2′-morpholine methylenebiphenyl-2-methyl formate methanesulfonate (IMH), a new drug for fatty liver treatment, was determined through differential scanning calorimetry (DSC). Analysis of two-factor non repeatability method was performed in the investigation the effects of two factors (heating rate and sample weight) on purity determination. The DSC experimental parameters were optimized as follows: heating rate was 10 ℃·min^-1, temperature range was 150-300 ℃, sample weight was 2.0-4.1 mg, and N₂ flow rate was 80 mL·min^-1. The linear correlation coefficient (r) of this DSC method was 0.999 8. The RSD value (n = 6) of precision was 0.03%. The standard value and uncertainty of the purity results of the multiple batches of IMH drugs were (99.74 ± 0.29)%, (99.91 ± 0.28)%, (99.90 ± 0.28)%, and (99.81 ± 0.28)% with inclusion factor (K) of 2 and confidence probability (P) of 0.95. The results were basically consistent with the results of the mass balance method. The DSC mehod is a simple, rapid and accurate method, and provides a new reference method for determining the purity of IMH drugs, improves the accuracy and reliability of purity determination.

Objective To discuss the short-term clinical effect including functional change of lipiodol- arsenic trioxide emulsion on the primary hepatic carcinoma.Methods Fifty-two patients undergone arterial chemoemblization were selected and then randomly divided into two groups:treatment group(n=27)and control group(n=25).Patients in treatment group were treated with lipiodol-arsenic trioxide,while those in control group treated with mitomycin,epirubicin,cisplatin or lipiodol.Clinical symptoms and six liver function parameters were observed and analized.Results The clinical symptoms of patients in treatment group improved much better than those in control group,and the liver function impairment of patients in treatment group also decreased more than those in control group.Conclusions Lipiodol-arsenic trioxide is an effective and safe intervention-therapeutic embolization material for primary hepatic carcinoma.

Objective To evaluate the safety of coronary intervention using iodixanol in patients with diabetic renal insufficiency.Methods Clinical data of 97 patients with diabetic renal insufficiency undergoing coronary intervention during June 2004 to June 2006 were retrospectively analyzed,50 of them with iodixanol,an iso-osmolar contrast medium (iodixanol group),and 47 with iopromide,a hypotanic contrast medium (iopromide group).Judkin's coronary angiography showed 167 diseased vessels in the patients,65 in anterior descending branches,44 in circumflex branches,and 58 in right coronary arteries. Levels of serum creatinine and blood urea nitrogen were determined before percutaneous coronary intervention (PCI),on the day of the procedure,the 3rd and 7th days after PCI,respectively,as well as radiocontrast media-induced nephropathy (CIN) was observed.Results Totally,192 drug-eluting stents were successfully implanted in 167 diseased vessels,and all patients' angina pectoris symptom disappeared soon after the procedure,with a full success.No acute or subacute stent thrombosis and major adverse cardiac events (MACE) occurred.Two patients (4%) in iodixanol group and 10 (21%) in iopromide group got CIN,with a statistical significance (P