Objective: To construct a lentiviral vector of RNA interference(RNAi) of TGF-?1 gene. Methods: Three target sequences were selected according to TGF?1 mRNA sequence firstly,then three pairs of oligonucleotide sequences according to these target sequences and one pair of negative control oligonucleotide sequence were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pRNA-U6/Lenti plasmid. After identified by restriction enzyme digestion and sequencing ,these plasmids were cotransfected along with ViraPowerTM Lentiviral Packaging Mix into 293FT cells for the package of lentiviral particles. Then the lentiviral vector particles were transfected into Hela cells and TGF-?1 expression in the transfected cells was assayed by Real-Time PCR and ELISA. Results: It was confirmed by digestion and sequencing that lentiviral vectors had the correct structure and could express high titer of virus. After transfected into Hela cells,TGF-?1 expression was decreased significantly by two of these lentiviral vectors at both the protein and mRNA level and the first vector had the best interefence efficiency. Conclusion: The lentivirus RNAi vector of TGF-?1 was constructed successfully.

Objective To explore the application value of blood lipid,blood glucose,liver function and serum uric acid in the diagnosis of fatty liver in elderly patients.Methods A total of 62 elderly patients with fatty liver in our hospital from April 2016 to October 2016 were selected as study group,and 62 healthy persons were selected as control group.All the subjects were drawn fasting venous blood 4 mL.Rate method was used to detected liver function indexes of total bile acid(TBA),gamma glutamyltranspeptidase(GGT) and aspartate aminotransferase(AST),alanine aminotransferase(ALT),total bilirubin(TBIL) levels.The hexokinase method was used to detected blood lipid indexes,including very low density lipoprotein cholesterol(VLDL-C),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C),cholesterol(TC),triglyceride(TG) levels.The peroxidase colorimetric method was used to detected blood glucose index including glycated hemoglobin(HbA1c) and fasting blood glucose(FBG) levels.MINDRAY BS-400 automatic biochemical analyzer produced in Shenzhen MINDRAY company was used to detect the serum uric acid level.Value of blood lipid,blood glucose,liver function,serum uric acid in diagnosis for fatty liver were made statistical comparison.Results The levels of TBA,GGT,AST,ALT and TBIL in the study group were higher than those in the control group,the level of VLDL-C,LDL-C,TG,TC in the study group were higher than those in the control group,the HDL-C level was lower than that in the control group,the levels of FBG and HbA1c in the study group were higher than those in the control group,the level of serum uric acid in the study group[(447.12±91.03)μmol/L] was higher than that in the control group [(353.41±70.32)μmol/L],the differences were all had statistically significant(P<0.05).The sensitivity(80.65%,79.03%,82.26%,77.42%) and accuracy(90.32%,88.71%,90.32%,88.71%) of blood lipids,blood glucose,liver function,blood uric acid alone in diagnosis of fatty liver were lower than the combined diagnosis(95.16%,96.77%),the differences were statistically significant(P<0.05).Conclusion Fatty liver patients have abnormal blood lipids and blood glucose,liver function and blood uric acid level,combined with the above indexes for diagnosis can improve the sensitivity and accuracy of fatty liver,and provide guidance for the intervention,is worthy of promotion.

Intervention measures of a nursing student with hepatitis B carrier and situational mental disorder are reported,including inducing emotional release,helping nursing students appropriately recognize and deal with diseases,and searching effective social and family support.Related issues are also reflected on these interventions,such as to implement in-time mental nursing,stress hepatitis B related health education,deal with the conflict and balance between right of privacy and informed consent,and widespread related laws.

Objective To clarify whether the intravenous administration of embryonic stem cells(ESCs)could home in on injured myocardium caused by acute myocardial infarction(MI)in rats and to improve the cardiac function afterwards,and to explore the possible effect of inflammatory factor TNF-?on it.Methods ESCs were cultured in vitro and transfected with green fluorescence protein(GFP).The model of heart failure post MI in rats was established by permanent ligation of left coronary anterior descend artery.After operation the rats were administered with 10~7 ESC via tail vein every other day for 6 days.The sham and MI without ESCs infusion groups served as control.Six weeks later,the hemodynamic measurement was employed to evaluate the cardiac function.Immunohistochemistry assay was used to detect the GFP expression in heart and other organs,and the cardiac specific protein Tropinin I(Tn I)expression in GFP positive spot of heart.Meanwhile,the co-culture in vitro of ESC and neonatal rat cardiomyocytes was used to observe the effect of overexpression of TNF-?in cardiomyocytes on ESC migration.Results Six weeks after MI,cardiac function was significantly improved in rats administered with ESC in comparison with those of MI group.Histological changes demonstrated that infused ESCs formed GFP-positive grafts in infracted myocardium.Alternatively,the positive immunostaining for Tn I was found in the area corresponding to GRP positive staining.In regard to other organs,only in spleen a few GFP positive cells were found.This would indicate that circulating ESC could translocate to infracted myocardium and to form cardiac tissue.Migration assay of ESC in vitro indicated that cultured cardiomyocytes with overexpression of TNF-aobviously enhanced the migration of ESCs in comparison with cardiomyocytes without transfection. Conclusion Intravenously infused ESC could home in on infarcted myocurdium and futher differentiate into cardiomyocytes which led to the improvement of cardiac function.In the setting of acute MI the homing mechanism Could be associated with locally released inflammatory factors such as TNF-?which may play as a chemotactic agent on circulating ESC.

Humans ; Leukemia ; Neoplastic Stem Cells

Acute Disease ; Adult ; China ; Humans ; Leukemia, Myeloid ; therapy

Even though chemistry is now in place that potentially allows high coupling efficiencies to be attained, successful coupling is usually a challenge when so-called "difficult sequences" is encountered in peptide synthesis. Some factors that affect the coupling efficiency have been discussed and related methods to overcome those obstacles have been introduced in present review. All suggestions proposed here are valuable and also feasible to improve the coupling completeness in both liquid-phase or solid-phase peptide synthesis.

Objective To summarize the strategies of nutritional support for patients with hyperlipidemic acute pancreatitis (HLAP). Methods The clinical data of six HLAP patients who were treated in our department from January 1998 to December 2009 were retrospectively reviewed. Results The disease conditions of all patients were improved after fasting and early nutritional support and lipid-lowering treatment. Three cases with severe HLAP received enteral nutrition via a nasojejunal tube. Another three patients with milder HLAP obtained low-fat diet and orally administered lipid-lowering drugs. The plasma triglyceride levels decreased from (25.30 ± 20. 48 ) mmol/L to (5.41 ±4.55) mmol/L. No relapse was noted during follow-up (range: 3 -24 months). Conclusions The approaches of nutritional support for HLAP patients should be customized based on the specific disease conditions,and the enteral nutrition and lipid-lowering drugs should be reasonably and carefully designed and applied. For severe HLAP, nasojejunal tube should be maintained in the early stage to ensure the application of enteral nutrition and lipid-lowering drugs, protect the integrity of gut mucosal barrier, and decrease the fluctuation of blood glucose.

Aged ; Aged, 80 and over ; Humans ; Leukemia, Myeloid, Acute

Objective To explore the effect and mechanism of osteoprotegerin on apoptosis in human umbilical vein endothelial cells(HUVECs) induced by triiodothyronine(T3) with different concentrations.Methods HUVECs were cultured in control group (0 nmol/L T3),low-concentration group (0.1 nmol/L T3),physiologicalconcentration group(1 nmol/L T3),high-concentration group(10 nmol/L T3) with OPG for 48 h.The cell apoptosis was measured by cytometry and Hoechst 33258.The expressions of p-IκKβ,lκKβ,p-TSC2,TSC2,p-S6K,S6K,p-AKT,AKT,Bcl-2,and Bax were analyzed by Western blot.Results (1) After OPG pretreatment and being cultured in control group、low-concentration group and high-concentration group,the HUVEC apoptosis was decreased significantly(P＜0.05) as compared with groups without OPG pretreatment.There was no difference in physiological concentration group before and after intervention of OPG.(2) The expression of p-IκKβ,p-S6K,and Bax in other groups were higher than those in physiological concentration group ; however,the expressions of p-TSC2,p-AKT,and Bcl-2 were lower than those in physiological concentration group(P＜0.05).After OPG pretreatment in control group,low-concentration group,and high-concentration group,the expressions of p-IκKβ,p-S6K,and Bax decreased significantly (P＜0.05)and the expression of p-TSC2,p-AKT,and Bcl-2 increased significantly (P＜ 0.05) ; there were no differences in the expressions of p-IκKβ,p-S6K,p-TSC2,p-AKT,Bcl-2,and Bax in physiological concentration group before and after intervention of OPG.Conclusions Low and high concentrations of T3 increase apoptosis of HUVECs by way of activation of IκKβ/mTOR pathway.Osteoprotegerin is able to protect HUVECs from apoptosis induced by low and high concentration of T3 via the way of inhibiting the IκKβ/mTOR pathway.