Objective To estimate the influence of 1,25(OH)2D3 on the proliferative ability of and methylation levels of genomic DNA and proliferation-associated gene promoter in human HaCaT keratinocytes.Methods Some cultured HaCaT cells were treated with 1,25 (OH)2D3 of 10-6,10-7 and 10-8 mol/L for 24 hours,then,methyl thiazolyl tetrazolium (MTT) assay was carried out to evaluate the proliferative activity of cells,and a global DNA methylation quantification kit was used to determine the global DNA methylation level.Real-time PCR was conducted to quantify the mRNA expression of DNA methyl transferases (DNMTs) and methyl-DNA binding domain (MBD) proteins,and methylation-specific PCR (MS-PCR) to evaluate the methylation status of promoter region in the programmed cell death 5 (PDCD5) and tissue inhibitor of metalloproteinase 2 (TIMP2) genes,in HaCaT cells after 24-hour treatment with 1,25 (OH)2D3 of 10-6 mol/L.The HaCaT cells receiving no treatment served as the control.Results Compared with the untreated HaCaT cells,those treated with 1,25(OH)2D3 of 10-6 mol/L showed significantly down-regulated proliferative activity (0.152 ± 0.027 vs.0.290 ± 0.017,P ＜ 0.01),global DNA methylation level (0.187 ± 0.071 vs.0.316 ± 0.049,P ＜ 0.05),DNMT3a and DNMT3b mRNA expression levels (P ＜ 0.01 or 0.05),but markedly upregulated mRNA expression levels of MECP2,MBD2,PDCD5 and TIMP2 (P ＜ 0.01 or 0.05).Moreover,the DNA methylation levels within the promoter region of PDCD5 and TIMP2 genes were significantly lower in HaCaT cells treated with 1,25 (OH)2D3 of 10-6 mol/L than in the control cells (0.38 ± 0.135 vs.0.72 ± 0.121,0.46 ± 0.172 vs.0.68 ± 0.133,both P＜ 0.05).Conclusions 1,25(OH)2D3 may down-regulate the global genomic DNA methylation level of,and modulate the expression of DNA methylationmodifying genes in,HaCaT cells.Furthermore,1,25 (OH)2D3 can decrease the promoter methylation levels but induce the overexpression of PDCD5 and TIMP2 genes,and decelerate the proliferation of HaCaT cells.

Objective To investigate the effects of endotracheal suctioning,turning over,oral caring and swallowingon cuff pressure,so as to provide evidence for the management of the endotracheal cuff.Methods During continuous monitoring of cuff pressure with pressure sensor,the changes of cuff pressure were recorded in the process of endotracheal suctioning,turning over,and oral caring.The data of cuff pressure were recorded including before activity,during activity,after activity for 5 min,15 min and 30 min.In addition,the data of cuff pressure were recorded including before swallowing,during swallowing,after swallowing for 1 min,5 min and 10 min.Results The cuff pressure during endotracheal suctioning and after endotracheal suctioning for 5 min was higher than that before endotracheal suctioning,the difference was statistically significant (P＜0.05);the cuff pressure during turning over and after turning over for 5 min was higher than that before turning over,the difference was statistically significant (P＜0.05);the cuff pressure during the oral caring was higher than that before oral caring,the difference was statistically significant(P＜0.05);the cuff pressure during swallowing was higher than that before swallowing,the difference was statistically significant(P＜0.05).Conclusion These clinical factors would lead to transient increase of cuff pressure including suctioning,turning over,oral caring,and swallowing.The instantaneous cuff pressure will mislead the staff to judge the safey of endotracheal cuff.The cuff pressure should not be blindly adjusted,so as to avoid the risks of leakage and aspiration.

Objective:Antimicrobial peptides ( AMPs ) are peptides generated in the biological defense system against exogenous pathogens ,which is all important constituent of the non-specific immune system.The immune system can react and sometimes play an important role in tumor control both in animal models and in humans.In this study,CecropinXJ were used in the BALB/c Eca109 cells-bearing mice model.To investigate whether CecropinXJ exert anti-tumor efficiency through regulating immune response when treated with tumor-bearing mice.Methods:The expression of cytokines and DC surface molecules were detected by immunohisto-chemistry and Flow cytometry.Results:The experiment of mice showed that CecropinXJ could significantly inhibit the proliferation of transplanted tumors in dose-dependent manner.There were significant difference between control group and 10mg/kg CecropinXJ group on the volume and weight of tumor ( P<0.05 ).CecropinXJ inhibited Eca 109 cells infiltrating growth in tumor tissue without affecting viscera,and the expressions of VEGF and bFGF proteins were higher than those in the control group .The expression of cytokines and DC surface molecules,the results that,compared with the control group,CecropinXJ could not significantly promote the expression of CD4,IL-4 and IFN-γ,while not involving the expression of surface molecules on DCs in spleen (P>0.05).In CecropinXJ treatment group,TNF-αlevel in serum was significantly higher than the control group ( P<0.01 ).Conclusion: CecropinXJ could significantly inhibit tumor growth in Eca109 cells-bearing mice,and might not be affected anti-tumor efficiency through regulating immune response.

Objective To investigate the protective effect of sodium tanshinone ⅡA sulfonate(STS) on adriamycin-induced H9c2 cell apoptosis. Methods H9c2 cells were divided into four groups: the control group, the ADR group, the STS group, and the AS(ADR+STS) group. MTT assay were used to detect cell proliferation and cell death. The cell cycle of H9c2 cells were determined by flow cytometry assay. Protein expression of Bcl-2, Bax and Caspase-3 was detected by Western blot assay. Results Compared with the control group, cell proliferation was significantly reduced , and the number of the cells was significantly increased in the G0/G1 phase in the ADR group(P < 0.01, P < 0.05). At the same time, the expression of Caspase-3 was dramatically enhanced and the ratio of Bcl-2/Bax was significantly reduced (P < 0.05, P < 0.01) in the ADR group. However, pretreatment of STS increased H9c2 cell proliferation, decreased the number of the arresting cells, inhibited the expression of Caspase-3, and improved the ratio of Bcl-2/Bax (P < 0.05, respectively). Conclusions STS can attenuate ADR-induced myocardial cell apoptosis.

Objective To review curative effect of coronary artery bypass grafting(CABG).Methods 10 patients are off-pump CABG at normal body temperature.The rest 222 were done with CPB under general anesthesia,hypothermia.Before operation cardiopulmonary function Was adjusted.During operation the function of heart,brain and renal Was carefully protected.After operation blood slucose was controlled,inflammation was prevented and pulmonary physical therapy was given to all patients.Results 226 patients discharged recovery and free of angina pectoris completely.3 patients with CABG died of low cardiac output after operation.1 patient died of effects of protamine allergy.2 patients died of multiple organ failure.Follow-up has been done in 211 patients for 2 months to 3 years,all leading normal life and working.Conclusion Careful and standardized preoperative and pefioperative treatment is one of the factors of SUCCeSSful coronary artery bypass grafting.

Objective To compare the diagnostic values of 3D CT and 2D CT imaging for tibial plateau fractures. Methods A retrospective study was performed to evaluate 21 tibial plateau fractures. Four experienced orthopaedists were asked To make assessmem of fracture classifications and characteristics using 3D CT and 2D CT images.Agreement test Was performed to evaluate interobserver and intraobserver reliability for fracture classification and criteria validity for fracture characteristics(intraoperative findings were taken as the golden standard).Furthermore,MeNemar test was used to compare the agreement of 3D CT results and 2D CT results respectively with the golden standard for fracture characteristics.Results When 3D CT images were used.interobserver reliability and intraobserver reliability for fracture elassiftcation increased to"almost perfect agreement".The agreement of golden standard with 3D CT results for fracture characteristics Was significantly higher than that with 2D CT results. Condmiou Since 3D CT imaging can improve the reliability and accuracy of preoperative assessment oftibial plateau fractures,it is helpful and valuable for tibial plateau fractures.

Objective To investigate the relation between positive fluid balance and the prognosis in severe sepsis patients with acute kidney injury (AKI).Methods A retrospective analysis of clinical data of 90 patients in our department was carried out.According to the final outcome,patients were divided into survival group (n =26) and non-survival group (n =64),in which the relation between patients' positive fluid balance and the prognosis was evaluated.Results (1) There was no statistically significant difference in the age and severity between survival group and non-survival group of patients with severe sepsis and consequent AKI,but the patients in non-survival group had greater volume overload.Compared with the survival group,higher mean fluid balance [(1 112.12±546.85) mLvs.(644.69±474.93) mL,P=0.00],and less urine output [(1 224.07 ± 708.79) mL vs.(2 032.36 ± 723.53) mL,P =0.00) in non-survival group.(2) There was no significant difference in mortality between early and late continuous renal replacement therapy (CRRT) during ICU care.However,the average daily fluid load in late CRRT patients was significantly greater than that in early CRRT patients [(1178.81 ±397.03) mLvs.(287.22 ± ± 433.53) mL,P =0.00] and the lung oxygenation index in late CRRT patients was significantly worse thanthat in early CRRT patients [(211.22±42.56) vs.(169.46±57.40),P=0.04] (3) The relevant variables to 28-day mortality in AKI patients with severe sepsis included CRRT treatment,oxygenation index and the average daily fluid balance ＞ 500 mL.Among them,fluid balance ＞ 500mL was an independent risk factor for AKI patients with severe sepsis.Their prognosis was worse if they had greater positive fluid balance.CRRT was the protective factor which could affect the prognosis of patients with severe sepsis complicated by AKI.Conclusions Patients with severe sepsis complicated by AKI has a high mortality.Persistent fluid overload can lead to increased mortality in patients with severe sepsis complicated by AKI.Early CRRT can reduce fluid retention in patients with renal failure and improve oxygenation index.

OBJECTIVE:To observe the effects of Breviscapine injection on serum copeptin,NT-proBNP and ischemia modi-fied albumin (IMA) level and hemorheology indexes in patients with acute cerebral infarction. METHODS:Data of 132 patients with acute cerebral infarction were selected and randomly divided into observation group(66 cases)and control group(66 cases). Control group received 1 Aspirin enteric-coated tablet,qd + 20 ml Muscular amino acids and nucleosides injection adding into 500 ml 0.9% Sodium chloride solution,intravenous infusion,qd + 4 ml Ozagrel injection adding into 250 ml Sodium chloride solution, intravenous infusion,qd. Observation group was additionally given 5 ml Breviscapine injection adding into 250 ml 0.9% Sodium chloride solution,intravenous infusion,qd. 7 d ag was regarded as 1 treatment course,it lasted for 2 courses. Copeptin,NT-proB-NP,IMA levels (showed by serum ACB value) and changes of related hemorheology indexes before and after treatment in 2 groups were observed. RESULTS:Before treatment,there was no significant difference in Copeptin,NT-proBNP,serum ACB val-ue,whole blood viscosity,plasma viscosity,hematocrit,ESR and fibrinogen levels in 2 groups(P>0.05);after treatment,Co-peptin,NT-proBNP,whole blood viscosity,plasma viscosity,hematocrit,ESR and fibrinogen levels in 2 groups were significant-ly lower than before,and observation group was lower than control group,while serum ACB value was significantly higher than be-fore,and observation group was significantly higher than control group,the differences were statistically significant(P<0.05). CONCLUSIONS:Based on conventional treatment,Breviscapine injection can significantly improve the copeptin,NT-proBNp and IMA levels,and improve hemorrheology.

BACKGROUNDTo evaluate the inhibitive effect of combination of 3TC with Ara-AMP against HBV in vitro.

METHODSUsed 2.2.15 cell as target cell. With radioimmunological technique and blot slot, the inhibitive effect of 3TC, Ara-AMP and the combination of both against the HBsAg, HBeAg and intracellular HBV DNA were investigated.

RESULTSThe inhibitive ratio of Ara-AMP against HBsAg, HBeAg was 45.48% and 41.46% respectively when its concentration was 400.0 microgram/ml. Although 3TC also has inhibitive effect in its experimental concentration, its effect is weaker. When Ara-AMP 50.0 microgram/ml was combined with 3TC 1.25 and 5.00 microgram/ml respectively, the inhibitive ratio against HBsAg were 19.92% and 17.32% respectively. Compared with using same concentration 3TC alone, the difference of results was significant (P<0.05). But when compared with using the same concentration Ara-AMP alone, the difference of results had no statistical significance (P <0.05). Remarkable inhibitive effects of combination of 3TC with Ara-AMP against HBeAg were n ot found. When 3TC 5.00 microgram/ml was combined with Ara-AMP 12.5 and 50.0 microgram/ml respectively, the inhibitive ratio against HBV DNA was 45.90% and 50.36% respectively. Comparing the content of HBV DNA in these groups with that of control group and the groups using the same concentration 3TC and Ara-AMP alone, the differences were significant (P <0.05).

CONCLUSIONSCombination of 3TC with Ara-AMP could enhance the inhibitive effects against HBV DNA.

Antiviral Agents ; pharmacology ; DNA, Viral ; drug effects ; metabolism ; Dose-Response Relationship, Drug ; Drug Synergism ; Hepatitis B virus ; drug effects ; Humans ; In Vitro Techniques ; Lamivudine ; pharmacology ; Vidarabine Phosphate ; pharmacology

Objective To investigate the prediction function of preoperative B-type natriuretic peptide in patients to receive on-pump coronary artery bypass grafting with postoperative complications. Methods One hundred and thirty-two patients , including 78 males and 54 females , received on-pump coronary artery bypass grafting from January 2013 to November 2014 and were enrolled in the study. The patients were (63 ± 11.35) years old ( range from 35 to 82 years). The level of BNP was determined before operation, after operation, and on day 1, 2, 3 and 7 post-operation. Relationships were analyzed between BNP and LVEF,ventilation time, length of stay in ICU, the need for inotropic agents or intra-aortic balloon pump (IABP), incidence of postoperative atrial fibrillation, and acute renal failure. Receiver operating characteristic (ROC) curve analysis was also performed to predict the role of BNP in postoperative complications. Result A negative correlation between preoperative BNP level and preoperative LVEF(r = -0.512,P < 0.05) was found. The preoperative BNP level was positively correlated with a series of adverse events. The preoperative BNP was used to predict incidence of postoperative atrial fibrillation , the possibility of using IABP , renal failure , length of stay in ICU exceeding 48h or mortality at 28 days, and the area under the ROC curve (AUC) was 0.780, 0.893, 0.818 and 0.820, respectively. Conclusion The preoperative BNP level is well correlated with the cardiac function before CABG , which may be a good predictor of postoperative complications after CABG.