BACKGROUND: Recent researches demonstrate that transplantation of bone marrow stem cells in the area of myocardial infarction can directionally differentiate into myocardial cells having normal physiological function and can promote newborn vascularization so as to repair infarction myocardium and improve injured cardiac function.OBJECTIVE: To observe short-term clinical effect of autologous bone marrow stem cell transplantation (ABMSCT) in percutaneous coronary artery on the treatment of acute myocardial infarction.DESIGN: Self-control study.SETTING: Department of Cardiology, Xiangtan Central Hospital.PARTICIPANTS: A total of 27 patients with acute myocardial infarction, including 16 males and 11 females, were selected from Department of Cardiology, Xiangtan Central Hospital from June 2004 to December 2006. Their ages coronary artery was finished after onset emergently; in addition, blood flow of infraction related vessels recovered to grade TIMI3. All patients provided the confirmed consent.METHODS: Operative procedure: All patients were performed with emergently interventional therapy of coronary artery after onset of acute myocardial infarction. One week later, percutaneous cavity tube technique was used to establish infarction related arterial pathway, and guiding filament was used to send micro-perfusion tube into stents. And then,separated bone marrow stem cell suspension was poured through central cavity of micro-tube into the distal end of infarction vessels. Operative evaluation: Dynamic electrocardiogram was evaluated for 24 hours before and after transplantation; in addition, left ventricular ejection fraction and myocardial perfusion defect scores were detected before and at 6 and 12 months after transplantation; otherwise, recovery state and complication were observed in follow up at 6 and 12 months after operation.defect scores: At 6 and 12 months after operation, left ventricular ejection fraction was higher than that before transplantation, and there was significant difference before and after transplantation (P＜0.05). While, myocardial perfusion defect scores were lower than those before transplantation, and there was significant difference before and arrhythmia were not found out, cardiac arrhythmia was not increased, and cardiac arrhythmia combining with malignancy not have any complications and in-stent constriction after operation.

Objective To investigate the correlation of the vascular endothelial growth factor (VEGF) gene variations in the promoter region with the sporadic Alzheimer' s disease (SAD) in Chinese Han population for better understanding the mechanism of SAD. MethodsThe polymorphisms of 279 SAD Chinese Han patients from Northern China were analyzed by comparing with those from 317 healthy individuals using the method of polymeraee chain reaction-restriction fragment length polymorphism ( PCR-RFLP) or direct sequencing.The commercial statistics package SPSS 11.5 was used to compare the distribution of the allele and the genotype, and to analyze their correlations with SAD. ResultsThree polymorphism sites were found for the VEGF promoters in the Chinese Han sample group including -2578C/A,- 2549I/D and- 1154G/A.- 2549I/D and- 2578C/A exhibiting strong linkage disequilibrium. Individuals with the A allele at position -2578 had an insertion of 18 nucleotides at -2459I/D, whereas CC homozygotes did not contain th es were found between the SAD patients and the controls in the 3 VEGF polymorphisms. After adjusting the data for gender, age and the ApoE ε4 allele using Logistic regression, the - 1154G/G genotype of the VEGF promoter might increase the risk of SAD in Chinese Han population.Among the subgroup without the ApoE ε4 allele, -2549D/-1154G haplotype might increase the risk for SAD (OR = 1.325, 95％ CI 1.023--1.716, P=0.033). ConclusionsThree polymorphism sites ( -2578C/A, -254911D, and -1154G/A) are found in the VEGF promoter regions in Chinese Han population. The-1154G/G genotype of the VEGF promote appears to increase the risk of SAD in Chinese Han population.In the absence of ApoE ε4, the -2549D/-1154G haplotype of the VEGF promoter appears to affect the risk for SAD.

Objective To explore the relationship between MR enhancement signs with MVD and VEGF expression in early breast carcer or benign lesion,improve the correct diagnosis rate of breast mass.Methods The MR enhancement signs(early enhmcement rate,eurve type of enhancement)of 31 patients with breast cancer and 19 benign lesions were analyzed retrospectively.Results Mean early enhancement rate of 31 neoplasm(20 invasive duet csncer,6 invasive lobule cancer,2 mucous cancer,3 situ lobule caner)were(68.4±14.7)%,enhancement curve type of which manifested as 18 type Ⅰ curve,11 typeⅡcurve and 2 type Ⅲ curve.Mean early enhancement rate of 19 benign mass(11 fibre adenoma,6 cystic hyperplasia,1 angioma,lobular tumor,1 infecmd cyst)were(47.6 ±15.2)%,of which 2 were typeⅠcurve,10 type Ⅱ cnrve and 7 type Ⅲ curve.In early cancer focus,mean MVD and VEGF expression index were 48.2±5.6 and 1.8 ±0.3,while in benign mass they were 30.8 ±6.1,0.9 ±0.3 respectivdy.Condusion Early enhancement rate,MVD and VEGF expression index in malignant focus were higher than those in benign mass,type Ⅰ curves were predominate in malignant lesions,by which early breast cancer could be diagnosed in time.

Objective To investigate the protective effects of atorvastatin on hyperphosphate-induced rat vascular smooth muscle ceils (RVSMCs) calcification and to discuss the mechanism. Methods RVSMCs were placed in various culture media, including normal phosphate medium, high phosphate medium, ZVAD-FMK medium and atorvastatin medium.Calcium content and cell protein content were quantified by the o-cresolphthalein complexone method and BCA protein assay respectively. Calcification was visualized by yon Kossa staining. And cell apoptosis was quantified by ELISA. Results (1)At day 3, 6, 9, RVSMCs calcification occurred more frequently in high phosphate medium than that in normal phosphate medium (P<0.05). (2)At day 6, RVSMCs calcification was significantly inhibited in 1.0 μmol/L and 2.0 μmol/LZVAD-FMK medium (P<0.05). And in 10 nmol/L and I00 nmol/L statin medium, RVSMCscalcium deposition significantly decreased (P<0.05). (3)RVSMCs apoptosis and calcification occurredfrequently in high phosphate medium. And atorvastatin significantly inhibited RVSMCs apoptosisboth in long-term and short-term (P<0.05). Conclusions Hyperphosphate can induce the calcium deposition of RVSMCs in vitro. Atorvastatin protects RVSMCs from phosphate-induced calcification by inhibiting apoptosis.

Insulin secretion and insulin resistance were evaluated in 152 subjects with normal glucose tolerance, impaired fasting glucose (IFG) and (or) impaired glucose tolerance (IGT). The data showed that subjects with IGT exhibited an evident deficit in the early and late phases of insulin secretion after glucose load and mild insulin resistance, and subjects with IFG demostrated evident insulin resistant and a great decline in early phase insulin response and HOMA-β.

BACKGROUND: Bone morphogenetic protein(BMP) is one of the most important cytokines that induce and promote seed cells to be transformed into osteocytes. Insoluble natural BMP can hardly affect the life of cultured seed cells. The expensive soluble recombinant BMP is also hard to work on the seed cells at the appropriate time and dose. Therefore, gene therapy technique provides us with a brand new idea of using gene-modified seed cells.OBJECTIVE: To transfect exogenous BMP-3 gene into the fibroblasts and screen the positive fibroblast clones that can express BMP-3 stably.DESIGN: Simple sample study.SETTING: Orthopaedic Research Institute, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: The fibroblasts(NIH3T3) were kindly presented by Professor Situ Zhen-qiang of the Stomatological College of Fourth Military Medical University of Chinese PLA.METHODS: This experiment was conducted in the Key Laboratory of Chinese PLA, which belongs to the Orthopaedic Research Institute of Fourth Military Medical University. BMP-3 gene was transfected into the fibroblasts through lipofectamin. The transfected cells were screened by G418. The separated cloned cells were identified through immunohistochemistry. The positively stained cells were the clones of BMP-3 expressing fibroblasts.MAIT OUTCOME MEASURES: The screening concentration of NIH3T3 cells, screening of positive transfected cells, and expression of BMP-3 in screened cells.RESULTS: BMP-3 gene was successfully transfected into the fibroblasts. BMP-3 expressing fibroblast clones were creened and identified through immunohistochemistry. Fibroblast strains with stable BMP-3 expression were obtained.CONCLUSION: The transfection of BMP-3 gene eukaryonic expression vector into the fibroblasts and obtaining of fibroblast strains with BMP-3 expression have laid foundation for the usage of gene-modified seed cells in future research of bone tissue engineering.

Oral glucose tolerance test(OGTT)was performed in 419 first-degree relatives(FDRs)of type 1 diabetes mellitus. GADA, IA-2A, and IAA were determined by radioligand assay, and the positive rates were 7.16%, 1.43%, and 1.26%, respectively. Intravenous glucose tolerance test(IVGTT)and nateglinide-OGTT were performed in 39 controls, 11 first-degree relatives with positive autoantibody(Ab+group), 14 ones with negative autoantibody(Ab-group)during 5-7 days.The first-phase insulin release(FPIR), area under insulin release during 0-10 min [AUC0-10] of IVGTT and the value of(ΔI30/ΔG30)of nateglinide-OGTT in Ab+group were lower than those of control and(2.75±0.37 vs 3.61±1.05)mU/mmol, all P＜0.05]. The 1st min insulin release in Ab+group was lower than that of Ab-group [(3.80±0.30 vs 4.52±0.70)mU/L, P＜0.05]. The HOMA-IR was higher in Ab-group than that in control group(2.92±1.04 vs 1.96±1.22, P＜0.05). The results suggest that the positivity rates of autoantibodies in FDRs of type 1 diabetes mellitus are very close to those of Caucasian. There exist insulin secretion defects in FDRs with positive autoantibody while insulin resistance in FDRs with negative autoantibody.

Objective To analyze the character of Pol-specific T lymphocyte responses and identify immunodominant region recognized in Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases. Methods Eleven Chinese HIV-1 recombinant subtype B/C infectors infected in 18 months, 25 which infected time more than 3 years and 10 HIV-1-seronegative healthy individuals were enrolled. HIV-1-specific T lymphocyte responses were analyzed by an IFN-γ ELISPOT assay against 249 overlapping peptides spanning HIV-1 Pol protein in the present study. Results Pol-specific T lymphocyte responses of IFN-γsecretion were identified in 8 (72.73%) out of 11 infectors infected in 18 months, the specific T lymphocytes are mainly targe-ted at six peptides which amino acid position from Pol 481 to 631 in reverse transcriptase region: Pol5581, Pol5582, Pol5587, Pol5609, Pol5610 and Pol5615. There was a negative correlation between the breadth of re-sponse and peripheral CD4+ T cell count (P=0.0212, r=-0.762) ; Responses were identified in 15 (60%) out of 25 chronic infectors, the specific T lymphocytes are mainly targeted at four peptides which amino acid po-sition from Pol 241 to 295: Pol5521, Pol5525, Pol5526, Pol5531 and another peptide: Pol5638 which amino acid position from Pol 708 to 722 in reverse transcriptase region. There was a positive correlation between the magnitude of Pol-specific IFN-γ secretion T lymphocyte responses and plasma viremia (P = 0.006 95 , r = 0.660) . None of the seronegative healthy individuals gave the positive responses. Conclusion Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases mainly recognized different re-gions of Pol.

Objective To analyze characteristics of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype B/C infectors. Methods 19 HIV-1 recombinant subtype B/C infectors infected within 1 year, 40 chronic infectors infected for more than 3 years were enrolled in this cohort study. Elispot assay was used to observe HIV-1 specific T lymphocyte responses in HIV-1 recombinant subtype B/C infectors. Results Nef-specific T lymphocyte responses of interferon-gamma secretion were identified in 15 Chinese HIV-1 recombinant subtype B/C infectors infected within 1 year. The specific T lymphocytes were mainly targeted at four peptides which span from Nef 83 to 135: EVA7081.1, EVAT081.5, EVA7081.6 and EVAT081.48. Responses were identified in 29(75. 2%) infectors with more than 3 years of infection and the specific T lymphocytes were mainly targeted at three peptides which span from Nef 63 to 101 : EVA7081.43, EVA7081.44, EVAT081.45, EVA7081.47, EVA7081.48 and EVA7081.49. The average magnitude of response in infectors with less than 1 year of infection was 284. 13 SFC/106 PBMC. The average magnitude of response in infectors with more than 3 years of infection was 152. 44 SFC/106 PBMC. There was a significant difference between the two groups (U = 91. 000, P = 0. 002). Conclusions HIV-1recombinant subtype B/C infectors at different stages of diseases (less than 1 year and more thank 3 years) can recognize central region of Nef. The magnitude of Nef-specific IFN-γ secretion T lymphocyte responses in this cohort gradually decrease with disease progression.

Objective To analyze character of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype C/B' and subtype B' infectors and to identify the common immunodominant re-gions recognized by these infectors. Methods Fifty-nine HIV-1 recombinant subtype C/B' infectors and 27 subtype B' infectors were tested by IFN-γ enzyme linked immunospot (ELISPOT) assay using HIV-1 C/B' Nef overlapping peptides. Results Nef-specific T-cell responses of IFN-γ secretion were identified in 44 (74.58%) out of 59 HIV-1 recombinant subtype C/B' infectors. Ten peptides, EVA7081.1,5, 6, 7,43, 44, 45, 47, 48, 49 were mainly recognized. Amino acid position was from Nef63 to 115 and 117 to 139. Twenty of 27 (74.07%) HIV-1 subtype B' infectors recognized peptides. EVA7081.1,2, 43 and 49 were mainly recognized. Amino acid position was from Nef 63 to 77 and 87 to 119. There was no correlation be-tween the Nef-specific IFN-production of HIV-1-specific T cells responses and viral load or CD4 T cell count in both subtype infectors. Conclusion The immunodominant regions, from Nef63 to 77 and 87 to 115 were recognized by both Chinese HIV-1 recombinant subtype C/B' infectors and subtype B' infectors. These re-gions could be used in design of vaccine.