BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are widely utilized as seed cells or carriers in bone tissue engineering and gene therapy.Thus,how to obtain BMSCs with high purity arose more attentions of researchers.OBJECTIVE:To observe the effects of different separation methods and culture conditions on biological characteristics of rabbit BMSCs.METHODS:BMSCs were obtained by whole bone marrow culture,density grand centrifugal and red blood cell lysis.At 48 hours after culture,the cell numbers were counted,the time of passage was recorded,in addition,the cell morphology was observed by phase contrast microscope,and the CD44 antigen expression was identified using flow cytometry.The 3rd and the 7th generation aging cells were cultured with DMEM-LG,MEM-HG,and DMEM/F12 culture medium.MTT and count cell plat were used to evaluate the growth of BMSCs.Phase contrast microscope was used to observe the morphological changes of aging cells.RESULTS AND CONCLUSION:BMSCs could be separated by each method.The adherent cells showed shuttle or multiple angle shapes,with rich cytoplasm,and positive for CD44 antigen.The more cell number and shortest primary culture time was presented in red blood cell lysis group(P ＜ 0.05).DMEM/F12 could promote the proliferation of quiescent cells.And the cells prevented the better viability.The method of read blood cell lysis improving the efficiency of BMSCs adherent is an effective method of extraction of BMSCs.DMEM/F12 could promote the proliferation Maybe,DMEM/F12 is more suitable for BMSCs.

Objective:Study on the possibility that human follicular dendritic cells(FDC) can increase HIV infection and to investigate the related mechanisms.Methods:Human FDC was isolated from tonsils and cultured with infected heterogenous pheripheral lymphocytes or cultured with infected hemogenous tonsilar T lymphocytes.HIV P24 antigen,HIV 1 env DNA and HIV 1 nef RNA were determined respectively with ELISA method,PCR or RT PCR method.Results:FDCs were cultured with infected heterogenous lymphocytes,the P24 antigen is 6 7 times higher than the control group(P

Objective: To investigate the intervention of Shiquan Dabu Decoction (SQDBD) in the protein expression of VEGF. Methods: 50 C57BL/6 mice inoculated with lewis lung cancer were randomly divided into five groups and treated by different drugs. The ratio of antitumorand and antimetastatic of each group were calculated after 15 days’ treatment. The expression of VEGF was detected by SABC immunohistochemical method. Results: Low, medium and high dosages of SQDBD had the function of antitumor and antimetastusi at different extent. SQDBD could inhibit the positive expression of VEGF protein. Conclusion: Inhibition of the expression of VEGF protein is possibly one of the mechanisms of SQDBD in antitumor.

Objective To investigate the change of CD8+T lymphocyte non-cytotoxic antivirus response(CNAR)in slow progressors infected by HIV.Methods Applying with density gradient and immunomagnetic beads methods to purify the CD4+T lymphocyte from the healthy person and CD8+T lymphocyte from HIV-infected individuals.The CD4+T cell was infected by HIV(SF-33)virus and cocuhured with CD8+T cell.The culture supernatant was collected and the p24 value was detected by ELISA method.Results Our study showed that the CNAR function decreased by turns of slow progressors(SP),typical progressors(TP),health control group and AIDS group.There was significant difference between groups(P<0.01).We found a significant positive correlation between the CIM+T cell ture count and the CNAR function.The virus load didn't statistically correlate with the CNAR function.Conclusion The CNAR function possibly protected the HIV-infected individuals from progression.

Objective To establish a new methodology of evaluating rehabilitation status of local muscle injured by measuring the impedance and phase information.Methods An evaluation method was set up,which take symmetrical parts of injured muscle as control group during the measurement.Impedance and phase values of injured muscle were extracted with local impedance measurement method before and after rehabilitation to evaluate muscle recovery.Results Injured muscle tissue impedance values decreased significantly.In the process of rehabilitation of injured muscle,the impedance and phase values of the injured muscles were gradually approaching that of healthy muscle,and then achieved basically the same as reference values after rehabilitation.Conclusions Local impedance measurement method may provide a new,non-invasive and convenient evaluation method for the recovery of injured muscle tissue,and it has good prospects for clinical application.

90%inasinglestep,andtheproteinyieldwas2.5mg/L.ConclusionPurifiedChsp60kDantigenisobtained,andtheantigenmightbeappliedtodetectantibodyinpatientsinfectedwithChlamydialtrachomatis,andwillcontributetostudytheroleofChsp60inimmunopathogenesis.

OBJECTIVE: Transplantation of stem cell, especially neural stem cell, has been proven effective in treating nervous system diseases in animal models,which provides new hopes for recovery of nerve functions in patients suffering from various nervous system diseases. Multiple studies have been conducted on the role of neural stem cells in repairing nerve functions, and several hypotheses have been proposed to explain how the stem cells or neural stem cells acts to recover nerve function.DATA SOURCES: We searched Medline for English articles from January 1997 to August 2003 with stem cell, neural stem cell, bone marrow stromal cell, stroke, ischemic injury, nervous system disease, and neurotrophic factor as the keywords. We also searched Wangfang database for Chinese articles with the same keywords from January 2003 to December 2004.STUDY SELECTION: The articles found in these two databases were primarily screened with the inclusion criteria as follow: the subjects should be animals or human; and the study should be the basic and/or clinical researches on(neural) stem cells.DATA EXTRACTION: Totally 72 English and Chinese articles were found in the two databases, among which 14 articles were closely associated with the present study and 10 were indirectly related. Eight articles were excluded for repetition in the contents. Finally 16 articles were included for analysis.DATA SYNTHESIS: The full text of the articles were reviewed and summarized. The results of these articles suggest that neural stem cells are valuable in treating cerebral vascular diseases, brain and spine injury, nerve degeneration diseases, and peripheral nerves diseases.CONCLUSION: Stem cells, especially neural stem cells, play active roles in treating nervous systeme diseases such as differentiation promotion, nerves nourishing, and nerve substitute.

Objective To evaluate the effects of isokinetic quadriceps training with different ranges of motion for improving mal-patellar tracking after total knee replacement (TKR). Methods Isokinetic quadriceps training was administered to 52 TKR cases, involving 66 knee joints with poor patellar tracking. The subjects were randomly allocated into 3 groups for isokinetic training with ranges of 0-30°, 30-60° or 60-90°, with 22 knee joints in each group. The angular velocity was set at 30°/second. X-rays with loading and with 45° knee flexion were taken before and after the training. The lateral patello-femoral angle,the patella index and the patello-femoral index were recorded along with the maximum torque ( MT), the total work ( TW ), and the average work(AW) in the three different ranges of motion. Hospital for Special Surgery (HSS) scores were also recorded. Results After 3 months of training, the average lateral patello-femoral angle, patella index, and patello-femoral index in the 0-30° training group were all significantly better than before the training. However, there was no significant difference in these measures for the 30-60° and 60-90° groups. After training average MT, TW and AW increased significantly in all groups, but the improvements were significantly correlated with the differences in lateral patello-femoral angles, patella indices and patello-femoral indices only for the 0-30°group. When compared with the baseline values, a statistically significant difference in HSS score after training was observed only in the 0-30° training group. Conclusions Isokinetic quadriceps training with a 0-30° range of motion can best realign patellar tracking after TKR, and best improve knee function.

Objective To investigate the level of B7-H1 and its counter-receptor PD-1 expression in mDC and different subsets of T lymphocytes in HIV infected individuals in China and to analyze the correlation between the level of B7-H1/PD-1 and disease progression, and to demonstrate that PD-1/PD-L1-dependent inhibition is operating in HIV infected patients.Methods Percentage of B7-H1 and PD-1 expression in mDC, CD+4 T cells and CD+8 T cells from thirty-six untreated HIV infected patients and 20 health controls were selected and detected by flow-cytometry, its correlations with CD+4 T cell absolute counts and plasma viral loads were analyzed.Results The percentage of B7-H1 expression in mDC, CD+4 T cells and CD+8 T cells (mean 15.21, mean 20.63, mean 13.5)were higher than that of health controls (all P<0.05).The percentage of PD-1 expression in CD+4 T cells and CD+8 T cells (mean 17.91, mean 19.21)were higher than that of health controls (P<0.05, P<0.05). The level of B7-H1 and PD-1 expression were inversely correlated with CD+4 T-cell counts(mDC+B7-H1+:r=-0.647, P<0.01;CD+4B7-H1+:r=-0.489, P=0.002;CD+8B7-H1+:r=-0.372, P=0.026;CD+4PD-1+:r=-0.374, P=0.025;CD+8PD-1+:r=-0.455, P=0.005) and positively correlated with HIV viral load(mDC+B7-H1+:r=0.662, P<0.01;CD+4B7-H1+: r=0.426, P=0.01;CD+8B7-H1+:r=0.531, P=0.001;CD+4PD-1+:r=0.362, P=0.03;CD+8PD-1+:r=0.380, P=0.022).Conclusion The level of B7-H1 and PD-1 expression was associated with HIV disease progression, which provides a useful marker to define disease progression of HIV infection.

Objective To study the alternations of regulatory T cells in early HIV infected patients and its association with disease progression.Methods Fifty-one untreated HIV infected patients were enrolled and divided into 3 groups according to their infection time and CD+4 T cell levels(30 early HIV infected patients,15 typical progressors,6 AIDS patients).Twenty normal controls were enrolled.There were no significant differences between the age and sex among four groups.Blood was drawn by venipuncture from each subject in EDTA tubes and the levels of CD+4 CD+25 Foxp3 + regulatory T cells were detected by FACSAria flow-cytometry.Spearman correlation was used to detect association between CD+4 CD+25 Foxp3 + regulatory T cells and the absolute CD+4 T cells,viral load and activation of T cells.Results The levels of CD+4 CD+25Foxp3+ regulatory T cells showed the tendency of increasing tendency from normal control to early HIV infected patients,asymptomatic HIV infected patients and AIDS patients.Early HIV infected patients was significantly lower than that in AIDS group [3.79(2.11 - 5.43) % vs 8.09(4.90 - 8.90) %,Z = - 2.29,P = 0.022].The levels of CD+4 CD+25 Foxp3 + Treg cells were associated with viral set point(r = 0.479,P =0.038) and inversely associated with CD+4 T cells(r = -0.455,P =0.011) and closely associated with HLA-DR expression on CD+3 T cells(r = 0.533,P = 0.002).Conclusions The ratio of CD+4 CD+25 Foxp3 +regulatory T cells of early HIV infected patients was significantly increased and associated with viral set point and CD+4 T cell counts,which indicate that alternation of regulatory T cell may be an important factor contributing to the disease progression in early HIV infection.