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 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.

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.

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

Objective To assess the diagnostic value of various preoperative imaging examinations for the point of the lateral femoral circumflex artery perforators through the deep fascia,in order to provide reasonable evidence for the design of location and protocol of anterolateral thigh perforator flap.Mehtods From September,2013 to December,2013,23 patients (including 1 patient with bilateral anterolateral thigh perforator flap) preparing anterolateral flap surgery were randomly divided into 4 groups(every group was 6 side).The patients in different groups were using different imaging techniques respectively,such as handheld doppler (HHD),Color doppler sonography (CDS),MDCT angiography (MDCTA),CDS and MDCTA united image location technology.The flap sizes varied from 8 cm × 5 cm to 28 cm × 12 cm.Moreover the coincidence rate between preoperative and intraoperative location of the perforator vessel was evaluated.Results Ali flaps were survived after the operation.The donor site were primarily closed while partial skin grafting was performed in 8 cases.The appearance and functional resuhs were satisfactory with following up for 3 to 9 months.The rates of coincidence between the preoperative and intraoperative location of perforating branches were HHD 61.53％,CDS 87.50％,MDCTA 85.71％,CDS and MDCTA united image location technology 93.33％ respectively.The HHD group showed significant decrease compared with the other groups (x2 =7.92,P ＜ 0.01; x2 =6.15,P ＜ 0.05; x2 =12.54,P ＜ 0.01,respectively).The CDS group had no statistically difference with the MDCTA group (P ＞ 0.05).The united CDS and MDCTA group showed significant increase compared with the other groups (x2 =4.22,P ＜ 0.05; x2 =3.94,P ＜ 0.05).Conclusion CDS and MDCTA united image location technology are more accurate for perforator preoperative location in auterolateral thigh flap,and should be widely used.

Objective To investigate the prevention of stroke-associated pneumonia (SAP) with Ramipril.Methods 404 acute stroke patients with primary hypertension were randomized into two groups.Group A received ramipril 2.5 mg once daily orally,and then were titrated up to 5 mg/d after a week.Group B received Candesartan 4 mg once daily orally,and then were titrated up to 8 mg/d after a week.The treatment course was 28 days.All patients received examination of Standardized Swallowing Assessment (SSA).The amelioration of swallow function and incidence of SAP after treatment were compared between the two groups.Results There were 10 patients suffering from SAP in group A and 21 patients in group B.The incidence of SAP was lower in group A(5.0％)than in group B(10.4％)(P＜0.05).The positive rate of SSA was 87.1 ％ in patients with SAP,higher than the total rate (47.5％).Dysphagia was improved in both groups after treatment.The rate of dysphagia improvement in group A and group B was 81.4％ and 66.3％ respectively.There was significant statistics difference in the positive case of SSA between two group (P＜ 0.05).Conclusions Orally treatment with ramipril is effective to prevent stroke-associated pneumonia and toimprove swallow 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.