Objective In this study, we try to understand the effects of microenvironment on the differentiation of mesenchymal stem cells (MSCs) by coculturing MSCs with mature cardiomyocytes or culturing MSCs in cardiomyocyte-lysate, in this study. Methods MSCs isolated from mature rats were either cocultured with cardiomyocytes isolated from new born rats with the ratio of 1 to 4, or cultured in the medium containing 4-fold cardiomyocyte-lysate obtained by repeated freezing and defrosting of rat myocardial cells. The morphology of MSCs under light microscopy were observed daily for 7 days and immunostaining against cTnT and CD31 was performed on the 7~ th day. MSCs cultured in ordinary medium were observed as the control. Results Both MSCs cocultured with cardiomyocytes and cultured in cardiomyocyte-lysate were differentiated into myogenic cells and expressed cTnT and CD31 at the 7th day of cultivation. The MSCs in the control group did not change in morphology and express cTnT or CD31. Conclusion Both myocardial cell coculturing system and cardiomyocyte-lysate system can be used to induce bone marrow MSCs to differentiate into cardiomyocyte-like cells and endotheliocyte-like cells.

Objective To investigate the meridians and acupoints selection in the treatment of post-stroke spastic paralysis with Tuina (Chinese massage). Methods The literatures about Tuina therapy for post-stroke spastic paralysis were retrieved. The frequency of meridians and acupoints used was counted and analyzed. Results 99 papers were collected, which involved in 226 acupoints and 1602 times of application. The selected acupoints were distributed in all the fourteen meridians and 64.5% (1033/1602, 606 on upper limb, 427 on lower limb) of them were on the limbs. The acupoints of the Yang meridians was 75.0% (1200/1602) and the specific acupoints was 71.7% (1148/ 1602). Conclusion The acupoint selected was basically focused on the local areas in Tuina for post-stroke spastic paralysis, assisted with the involved meridians and distal acupoints. The acupoints of the Yang meridians were the first option, mainly on the extremities. The specific acupoints were the major components of the prescription, especially the Five-shu acupoints and the He acupoints.

Objective To identify potential linkage of cholesterol efflux with the expressions of ATP-binding cas-sette receptor A1 (ABCA1), ABCG1 and scavenger receptor B1 (SR-B1) in monocytes derived macrophages of patients with type 2 diabetes mellitus. Methods and Results Blood was collected from subjects with or without type 2 diabetes mellitus. Peripheral blood monocytes were differentiated for 72 hours into macrophages, and cholesterol efflux assays, Real-time quantitative PCR and western blot were performed. Macrophages from patients with type 2 diabetes mellitus showed a reduction in cholesterol efllux. The mRNA and protein expressions of ABCG1 in macrophages from patients with type 2 diabetes mellitus were significantly reduced. In contrast, the expression of ABCA1 and SR-B1 was not significantly different in both control subjects and diabetic patients. In addition, cellular cholesterol efflux from macrophages to autologous serum and pool serum was significantly correlated with the expression of ABCG1. Conclusion ABCG1 expression and cholesterol efflux are reduced in patients with type 2 diabetes mellitus. This impaired cholesterol efflux significantly correlates with decreased expression of ABCG1.

Objective To examine the effects of high-density lipoprotein(HDL) and lipoprotein-deficient serum(LPDS) isolated from patients with type 2 diabetes mellitus on cholesterol efflux through human skin fibroblast(HSF) and human hepatoma cell line(HepG2).Methods and Results Blood was collected from 13 patients with type 2 diabetes mellitus and 17 healthy volunteers,HDL and LPDS were isolated.Cholesterol efflux assays,RT-PCR and Western blot were performed with HSF and HepG2 cells.The HepG2 cells showed a high expression of scavenger receptor B1(SR-B1) and lack of functional ATP-binding cassette receptor A1(ABCA1) and ATP-binding cassette receptor G1(ABCG1) while HSF cells express SR-B1 at very low level and have a high expression of ABCA1 pretreated with 22-OH cholesterol.The cholesterol efflux from HepG2 cells to HDL isolated from patients with diabetes decreased significantly as compared to controls.However,cholesterol efflux from HSF cells to LPDS was not different between groups.Conclusion The function of HDL involving cholesterol efflux in type 2 diabetes mellitus was impaired while cholesterol efflux induced by LPDS from HSF cells was maintained,suggesting that HDL plays a critical role in mediation of intracellular cholesterol accumulation and progression of atherosclerosis inpatients with type 2 diabetes.

Objective To explore the mechanisms responsible for different coronary artery lesions with involvement of nitric oxide(NO),endothelin-1(ET-1),matrix metalloproteinases-9(MMP-9) and the matrix metalloproteinase inhibitor-1(TIMP-1).Methods The cases undergone coronary anography were collected and divided into three groups:group A,30 patients with the coronary artery ectasia(7 cases of simple coronary artery ectasia;18 cases of the coronary artery ectasia coexisting a small amount of plaque);group B,38 patients with coronary atherosclerosis;group C,32 patients with with normal angiograph(14 cases of coronary artery completely normal;18 cases with a small amount of coronary plaque only).Plasma NO,ET-1,MMP-9 and TIMP-1 level were measured by ELISA method.Results There are significant differences among three groups on NO level,MMP-9 levels,NO/ET-1 and MMP-9/TIMP-1(P

Objective To identify potential linkage of cholesterol efflux with the expressions of ATP-binding cassette receptor A1(ABCA1),ABCG1 and scavenger receptor B1(SR-B1) in monocytes derived macrophages of patients with type 2 diabetes mellitus.Methods and Results Blood was collected from subjects with or without type 2 diabetes mellitus.Peripheral blood monocytes were differentiated for 72 hours into macrophages,and cholesterol efflux assays,Real-time quantitative PCR and western blot were performed.Macrophages from patients with type 2 diabetes mellitus showed a reduction in cholesterol efflux.The mRNA and protein expressions of ABCG1 in macrophages from patients with type 2 diabetes mellitus were significantly reduced.In contrast,the expression of ABCA1 and SR-B1 was not significantly different in both control subjects and diabetic patients.In addition,cellular cholesterol efflux from macrophages to autologous serum and pool serum was significantly correlated with the expression of ABCG1.Conclusion ABCG1 expression and cholesterol efflux are reduced in patients with type 2 diabetes mellitus.This impaired cholesterol efflux significantly correlates with decreased expression of ABCG1.

Objective To explain the effects of C-reactive protein(CRP) on lectin-like oxidized low density lipoprotein receptor-1 expression on THP-1 derived macrophages and the related signal transduction pathways.MethodsTHP-1 cells were differentiated into macrophages with the stimulation of PMA.THP-1 derived macrophages were incubated with CRP and co-incubated with inhibitors of NF-?B、AP-1 and MARK signal transduction pathways.The expression of LOX-1 antigen and mRNA was analyzed by ELISA and RT-PCR.Results CRP stimulated the expression of LOX-1 antigen and mRNA on macrophages in a dose-dependent manner.NF-?B inhibitor BAY11-7085 suppressed the inducible effects of CRP on LOX-1 expression.Conclusion CRP increased LOX-1 expression on THP-1 derived macrophages at transcription and post-transcription levels.The NF-?B signal transduction pathway may be involved in such process.

Objective To investigate the potential change of cellular cholesterol efflux and the functional changes of ABCA1 on macrophages from diabetes animals. Methods High energy diet was given to golden hamster to make animal model of hyperlipidemia. Diabetes was induced by a single intraperitoneal injection of STZ (30 mg/kg). Macrophages were isolated and incubated with apoA-I or 8-br-cAMP in vitro. Cellular cholesterol content was measureal before and after treatment by HPLC. Results Intracellular cholesterol content of diabetic animals was higher than that in high energy diet and normal controls. Expression of ABCA1 mRNA was upregulated in diabetic group after incubation with apoA-I and cAMP. When macrophages were incubated with apoA-I, cholesterol efflux increased with the prolongation of incubation time.The amount of intracellular cholesterol efflux in 3 groups of animals showed the following relationship: diabetic animals exceed high energy diet fed animals, and the latter exceeds normal animals. Conclusion Intracellular lipid efflux depends mainly on the presence of extracellular apoA1 as well as membrane ABCA1 protein. The deposition of cholesterol in the macrophage of diabetic and insulin resistant animals may be related to the characteristic changes of quantity and function of apoA1 in these animals.

Objective: To explore the impact of knocking out wildtype p53 phosphatase 1 gene on heart function with the changes of cardiac tissue mRNA and protein expressions in experimental mice. Methods: Our research included in 2 groups: Wildtype (WT) mice group and Wip1 knockout (Wip1-KO) mice group. n=10 in each group. The heart function, ratio of heart weight to body weight (HW/BW) were examined and compared between 2 groups; cardiac tissue morphology was observed by HE staining; mRNA expressions of ANP, BNP, MCP-1 andα-SMA were determined by RT-PCR and protein expressions of Bcl-2, Bax and c-caspase3 were measured by Western blot analysis. Results: Compared with WT mice group, Wip1-KO mice group showed decreased Wip1 mRNA expression,P<0.05, decreased LVEF, LV fraction shortening and increased left ventricular end systolic diameter (LVESD), allP<0.05; Wip1-KO mice group had reduced BW and elevated ratio of HW/BW, bothP<0.05 even the heart weight was similar between 2 groups. There was no difference in cardiac tissue morphology between 2 groups; mRNA expressions of ANP, BNP, MCP-1 and α-SMA were similar between 2 groups,P>0.05; apoptosis related protein expressions of Bax/Bcl-2 and c-caspase3 were similar between 2 groups,P>0.05. Conclusion: Wip1 gene knockout may impair the heart function in experimental mice, while the relevant mechanism should be further investigated.