AIM:There is little evidence proving the molecular mechanism of WenxinKeli ( WXKL) .This study tried to explore the gene ex-pression profile and pathology alteration of WXKL-treated rabbits with myocardial infarction .METHOD: Twenty male adult rabbits were randomly divided into 4 groups:sham, model, WXKL and captopril groups .Model, WXKL and captopril groups underwent the ligation of the left anterior descending coronary artery , while sham group went through an identical procedure without ligation .WXKL (817 mg? kg-1? d-1), captopril (8 mg? kg -1? d-1) and distilled water (model and sham) were administered orally to the rabbits. 4 weeks later, hearts were taken out for expression chip and pathological staining (HE, Masson and TUNEL) after echocardiography. RESULT:WXKL could down-regulate genes associated with inflammation (CX3CR1, MRC1, and FPR1), apoptosis (cathepsin C and TTC5) and neuro-hormonal system (ACE and EDN1), and up-regulate angiogenesis promoting gene like RSPO 3, which explained why WXKL group represented with better cardiac function , less histopathological injury and slighter apoptosis .CONCLUSION:WXKL plays an important role in suppressing inflammation , inhibiting renin-angiotensin system and alleviating apoptosis , and might be a promising Chinese medicine in treating patients with myocardial infarction .

BACKGROUND: It has been reported that type 1 diabetes mellitus can result in impairments of bone regeneration and repair, and local injection of fibroblastic growth factor-2 (FGF-2) can obviously promote fracture healing, but its effect on type 2 diabetes mellitus is still unclear.OBJECTIVE: To observe the effects of recombinant human fibroblastic growth factor-2 (rhFGF-2) combined with soluble tumor necrosis factor receptor-1 (sTNF-R1) on impaired bone regeneration and repair in type 2 diabetes mellitus.DESTGN: A randomized controlled trial.SETTING: Department of Orthopaedics, the Third Xiangya Hospital of Central South University.MATERIALS: Twenty male Zucker diabetic fatty rats (ZDF/Gemi-fa/fa), 10 weeks of age, were purchased from Charles River Laboratory. rhFGF-2 was obtained from Orquest Incorporation; sTNF-R1 protein (PEG-r-metHu-sTNF-R1) was provided by Amgen Incorporation.METHODS: This experiment was finished in the central lab of the Third Xiangya Hospital of Central South University from September to November in 2006. ①Grouping: The 20 rats were randomly assigned into treated group (n =10)and control group (n =10). ② Experimental methods: All rats were examined for body mass, blood glucose, glycosuria and glycosemia. One week later, all the rats underwent the standard DO protocol, including placement of the external fixators and osteotomies to the left tibia. Distraction was initiated in the following morning (one day latency) at 0.2 mm b.I.d. (0.4 mm per day) and continued for 14 days. During surgery, all the rats received an injection of either rhFGF-2(25 mg/kg) for the treated group, or physiological saline (25 mg/kg) for the control group, into the hematoma of the osteotomic gap. The sTNF-R1 (8 mg/kg) or the same. Amount of saline was subcutaneously injected into the treated and control rats respectively every other day for 14 days. Evaluation: The serum biochemical indexes, amount of bone formation and number of proliferative cells in the distraction gaps were determined.MAIN OUTCOME MEASURES: Biochemical indexes, amount of bone formation and number of proliferative cells in the distraction gaps.RESULTS: All the 20 rats were involved in the final analysis of results. ①The blood glucose, glucosuria, ketonuria,serum levels of insulin and osteocalcin were not obviously different between the treated group and control group (P ＞0.05). ② The area and density of mineralization of the distraction gaps, and the endosteal and peristeal new bone formation in the treated group were all obviously higher than those in the control group (P ＜ 0.05-0.01). The number and percentage of the positive cells of proliferating cell nuclear antigen (PCNA) in the distraction gaps were obviously higher in the treated group than in the control group (P ＜ 0.05-0.01).CONCLUSION: The local application of rhFGF-2 combined with sTNF-R1 can enhance bone formation by increasing the proliferation during distraction osteogenesis in ZDF rats. The combination of rhFGF-2 and sTNF-R1 may be an effective treatment for type 2 diabetic patients with fracture healing problem.

Objective:To observe the dynamic change of cystathionine β-synthase during cerebral ischemia-reperfusion and its effect in rats.Methods:The ischemic model was established with line embolism to block the middle cerebral artery .The reverse tran-scription-polymerase chain reaction ( RT-PCR) and Western blot assay were used to assess the expression of cystathionine β-synthase (CBS) in SHAM group,I group,and IR group.ELISA assay was performed to detected the homocysteine (HCY) level in plasma.After treating with the inhibitor of cystathionine β-synthase called hydroxyla mine(HA),the expression of hemeoxygenase 1(HO-1) and the pathologic change of the brain was evaluated .Results:As compared to sham group ,the expression of CBS was significantly up-regulated in ischemia-reperfusion group at 12 h post-reperfusion.Meanwhile,it existed the lowest level of HCY at 12 h post-reperfusion,comparing to sham grouzp ( 5.73 ±1.17 vs 2.88 ±0.93 , F=25.56 , P=0.001 ) .When inhibited the activity of CBS via using HA , the down-regulation of HO-1 protein and further damage in neuron were observed .Conclusion:Cystathionine β-synthase serves as an protective factor during cerebral ischemia-reperfusion.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are few in bone marrow, and they are easily mingled with other cells, especially red blood cells. Therefore, intervention needs to be depleted in the process of isolation of red blood cells so as to obtain highlypurified BMSCs as many as possible.OBJECTIVE: To isolate and culture BMSCs of rats with red blood cell lysis method, and perform biological identification.DESIGN: Observation and controlled trial.SETTING: Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts.MATERIALS: Fifty 30-day-old male SD rats, weighing about 100 g, of SPF degree, were purchased from Beijing Experimental Animal Center (License No. SCXK (Jing) 2002-0003) and involved in this trial. DAB condensed chromogen (hydrogen dioxide included, Zhongshan Company), rabbit anti-ret polyclonal antibody (Boster Co.,Ltd., Wuhan), fetal calf serum (PAA, Austria) and LG-DMEM medium (Sigma Company, USA) were used in this trial.METHODS: This trial was carried out in the Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts during September 2004 to September 2005. The rats were sacrificed by dislocation to expose bone marrow cavity. Cell suspension was collected. BMSCs were isolated and cultured primarily by whole bonemarrow culture method. ① Red blood cells lysis test: A, B, C and D 4 tubes were chosen and filled with 0.5 mL bonemarrow rinse solution which was filtered and fully beat upon. Then, red blood cell lysis buffer of 2 mL, ammonium chloride of 2 mL, phosphate buffer normal saline of 2 mL and 0.04 volume fraction acetic acid of 0.5 mL were correspondingly added into the 4 tubes. In each tube, absorbance and hemoglobin concentration were measured and cell growth was observed. ② Observation of growth curve, doubling time and surface marker molecule expression of BMSCs: Based on the formula, population doubling time (TD) =t[lg2/(lgNt-lgN0)] (NO and Nt represented the cell number after inoculation and t hours after culture ,respectively), cell population doubling time was calculated and traced and growth curve of the 2nd, 4th and 6th generations of BMSCs were analyzed; The proliferation of BMSCs was measured by methylene blue staining method; Surface marker molecule expression of BMSCs was detected with immunocytochemical staining.MAIN OUTCOME MEASURES: ①Observation of isolation and culture of bone marrowmesenchymal stems of rats. ②Effect of different methods on the lysis of red blood cells and the growth of BMSCs. ③ The growth curve and cell doubling time of the 2nd, 4th and 6th generations of cells. ④Surface marker molecule expression of BMSCs of rats.RESULTS: ① Results of isolation and culture of BMSCs of rats: After 48-hour primary culture, most of the cells had adhered to the wall, and 72 hours later, division growth of the adherent cells presented. Seven to eight days later, cell colonies formed obviously, and then increased quickly, expanded incessantly and fused with each other. On 14 to 16 days, cell clones grew densely. Immediately generative cells presented ball-shape, subsided and adhered the wall verysoon. Some few round cells suspended. Adherent cells distributed evenly and proliferated quickly within 3 to 5 days. Although cell morphology of generative cells did not change after passage, cell proliferation was speeded up obviously and cells covered the bottom of the whole bottom on about 6 days. ② Effect of different methods on red blood cell lysis and the growth of BMSCs: hemoglobin concentration in the red blood cell lysate-treated group, ammonium chloride-treated group and 4％ acetic acid-treated group was significantly higher than that in the phosphate buffer normal saline-treated group, with significant difference (P ＜ 0.01). ③ Observation of growth curve of different generations of BMSCs: The growth curves of the 2nd, 4th and 6th generations of the cells were basically the same: latent period about 1 to 2 days, then logarithmic growth phase, peak on the 5th day and finally plateau phase (about on the 5th to 7th days). The latent period of the 6th generation of BMSCs was not obvious and the population doubling time of BMSCs was about 34 hours. ④ldentification of immunophenotype of different generations of BMSCs: Both CD44 and CD106 staining of each generation of cells were positive, presenting brown granule sediments, and CD34 staining was negative.CONCLUSION: Inoculation of red blood cells lysate-treated bone marrow rinse solution can boost the adherent rate of BMSCs, and does not influence its post-adherent growth, so it is a feasible separation method. Cell surface marker staining confirms that thecells isolated in this study are BMSCs.

Objective To observe the expression of free Ca2+ and Angiotensin Ⅱ 1 type receptor(AT1 R)in vascular smooth muscle cells(VSMCs)of patients with hypertensive or normotensive coronary artery diseases(CAD).Methods During the coronary artery bypass graft operation,the surplus saphenous vein of patients who admitted to our Cardiac Surgery Department was collected and cultured.All patients were divided into hypertensive or normotensive group.Free Ca2+ in the cultured human VSMCs was determined by confocal laser scanning microscope(CLSM)after different kinds of AngiotensinⅡ being added,respectively.Total RNA was extracted from cultured VSMCs.Then RT-PCR was conducted for the observation of the expression of AT1R in both groups.Results Ca2+in human VSMCs rapidly increased when stimulated by Angtensin Ⅱ in two groups.After stimulated by Angiotensin Ⅱ,both free Ca2+ level and the expression of AT1R in VSMCs of hypertensive patients were higher than those of the normotensive patients(P＜0.05).Conclusion There are certain changes of free calcium in the cultured human vascular smooth muscle cells,when stimulated by Angiotensin Ⅱ.There are also difierences in AT1R expression between hypertensive CAD patients and normotensive CAD patients.

Two hundred and fourteen patients with non-ST-segment elevation acute coronary syndrome (NSTEACS) underwent percutaneous coronary intervention (PCI).Serum ischemic modified albumin (IMA) levels were measured in patients at admission.The major adverse cardiac events (MACE),including cardiac death,nonfatal myocardial infarction (MI) and recurrent ischemia leading to urgent revascularization were observed during 1-y period of follow-up.Receiver operating characteristic (ROC) curves,Kaplan-Meier analysis and Cox regression were used to assess the prognostic value of IMA for 1-y MACE.Twenty one patients experienced major adverse cardiac events during 1-y follow up period,including 6 cases of cardiac death,8 cases of new or recurrent MI,7 cases of target vessel/lesion revascularization or coronary artery bypass grafting (CABG).ROC showed that the area under the ROC curve (AUC) was 0.667,and when IMA was used to predict 1-y major adverse cardiac events,the cut-off value of 65.3 kU/L was most effective.Kaplan-Meier analysis showed that IMA was significantly correlated with the occurrence of 1-y MACE(P ＜ 0.01).But Cox regression model showed that IMA levels were not independent risk factor for 1-y MACE in NSTEACS patients,when adjusted with other risk factors.

Exercise ; Heart Arrest ; Humans

Objective: To study the predictive value of 12-lead electrocardiogram (ECG) abnormality on prognosis of chronic heart failure in patients with dilated cardiomyopathy (DCM-CHF).
Methods: A prospective, multicenter follow-up study in 787 DCM-CHF patients was conducted, and the endpoints were obtained by clinical visit, mail contact and telephone conversation. The independent predictors for all cause death were determined by Cox regression analysis, QRS duration > 120 ms was studied and the survival rates were investigated by Kaplan-Meier analysis.
Results: There were 203 patients died during the follow-up period. Cox regression analysis found that the following indexes were related to all cause death: atrial fibrillation (AF) (HR=2.064, 95% CI 1.102-3.864,P<0.05), non-sustained ventricular tachycardia (NSVT) (HR=3.887, 95% CI 1.554-9.724,P<0.05) and QRS duration (HR=1.010, 95% CI 1.002-1.018, P<0.05). Kaplan-Meier analysis revealed that the survival rates were different by each stratiifcation of QRS duration,P<0.05.
Conclusion: ECG indexes of AF, NSVT and QRS duration had the important impact on the survival rate in DCM-CHF patients; there were signiifcant differences between QRS durations and survival rates.

BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic.
OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro.
METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope.
RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.

Objective To explore renin expression in cytomegalovirus(CMV) infected juxtaglomerular cells(JG) and its biological significance. Methods JG model cell line As4.1 cells derived from kidney tissue were respectively incubated with murine CMV at multiplicities of infection(MOI) of 10, 0.1 and 0 for 5 d, and the control was mock infection with the same amount of ultraviolet inactive CMV as MOI 10, then the cells were harvested. CMV immediate early gene(IE1) mRNA in the cells was tested by RT-PCR. The renin positive cells and the renin fluorescence granules in the cells were examined by immunofluorescence stain. Whether or not re-nin antigen and CMV antigen were showed in the same cells by FITC and TRITC immunofluorescence. The renin gene expression in the cells was individually detected by real-time RT-PCR and Western blot. Results The cells infected by CMV showed typical cytopathic effect(CPE) and viral plaques in the cell monolayer. CMV IE1 mRNA was found in the viral infected cells by RT-PCR. The mass or ring granules of renin positive fluorescence appeared in the cytoplasm of the CPE cells. The renin positive cells congregated around the viral plaques. Renin positive granules and CMV positive granules showed in the same cells. Renin expression in the CMV infected cells exhibited in a dependent manner of ratio of infectious virus particles to cells. Conclusion CMV infection of the cells derived from kidney tissue induces renin expression related to a new pathogenesis of cardiovascular diseases.