Association between IFN-γexpression and pathological damage of lung tissue during pulmonary tuberculosis / 实用医学杂志

Objective To investigate the relationship between the expression of IFN-γ, IL-2 and TNF-αand pathological damage of lung tissue, and to provide reference values for protection and treatment of pulmonary tuberculosis. Methods BALB/c mice models of pulmonary tuberculosis were established by infection with Mycobacterium tuberculosis H37Rv through the caudal vein. At 0 (not infected),2,4,6W and 8W after infection, the bacterial loads in the lung were performed and the pathological damage of the lung was evaluated, respectively. At the same time, the expression of IFN-γ,IL-2 and TNF-α-positive cells in the lung tissues was detected by immunohistochemistry. Results Two weeks after infection, the expression of IFN-γ, IL-2 and TNF-αpositive cells was obviously increased compared with those at 0 week. The proportion of IFN-γ-positive cells were obviously decreased at 6 weeks (9.25%) to 8 weeks (5.67%) after infection with Mycobacterium tuberculosis H37Rv. At the same stage, there is a limited decrease tendency of TNF-α and IL-2 expression compared with those at 4 weeks after infection, and there was no significant difference compared with those at 2W or 4W after infection. At 8 weeks after infection, the bacterial loads (8.43) in the lung and the scores(17) of pathological damage were significantly increased compared with those at 6 weeks after infection. Conclusion The high expression of IFN-γ, IL-2 and TNF-αprotect lung tissues against MTB infection at the early stage. The decrease of IFN-γexpression is responsible for serious damage of the lung tissues in the persistent infection.

Study on immunogenicity elicited by a recombinant vaccine of rBCG-Rv3133c to fight against dormancy Mycobacterium tuberculosis / 生物医学工程学杂志

To obtain a vaccine to defend from dormancy Mycobacterium tuberculosis, we constructed the recombinant Bacilli Calmette-Guérin (BCG) vaccine with Rv3133c encoding dormancy-correlated transcriptional regulatory protein DosR in Mycobacterium tuberculosis as a target gene, and evaluated its immunogenicity in BALB/c mice. In this study, we constructed the recombinant plasmids of rpMV361-Rv3133c using gene colon technology. We then transformed BCG strains with above-mentioned plasmids to obtain recombinant vaccine of rBCG-Rv3133c. We used the rBCG strains successfully constructed to vaccinate in BALB/c mice. 30d and 180d after immunization, the specific antibody titers were determined to investigate humoral responses induced by recombinant vaccine. We detected changes of splenocyte subsets of CD4+T, CD8+ T cells and cytokine of IFN-gamma secreted by splenocytes for evaluation of cellular immune responses. The results showed that the rBCG-Rv3133c was able to induce higher levels of antibody titer, stronger proliferative responses and higher IFN-gamma production comparing with BCG vaccine. The results also suggested that this recombinant vaccine was a more efficacious tuberculosis vaccine for further study.

Differentiation of rat bone marrow stromal stem cells into neuron-like cells induced by breviscapine / 解剖学报

Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSCs) in vitro by breviscapine,and to offer reference for the applying of BMSCs. Methods The bone marrow stromal cells of SD rat were separated and purified by passsage culture.After phenotype characterization,the 4th passage of BMSCs was exposed to breviscapine. Differentiation was observed under phase contrast microscope every 6 hours and the cells were stained immunocytochemically with neuronspecific enolase(NSE)and glial fibrillary acidic protein(GFAP).The induced cells activity was detected by MTT. NSE and GFAP were determined by flow cytometry and RT-PCR. Results The BMSCs were CD44~+,CD54~+,CD34~-. After 18 hours of induction, cytoplasm of BMSCs partly contracted with protruding;The immunocytochemical staining was performed on cells after induction for 24 hours,the rates of NSE and GFAP staining positive were(48.7±3.4)% and(56.8±4.2)%.By using flow cytometer, expressions of NSE and GFAP showed much higher in induced group than that in control group. By using RT-PCR, expressions of NSE and GFAP were positive in induced group. Conclusion Breviscapine could induce adult rat BMSCs to differentiate into neuron and glial cells.

Repair of sciatic nerve defects with tissue engineered nerves constructed with marrow stromal cells / 中国组织工程研究

BACKGROUND: Schwann calls are seed cells for constructing tissue engineered peripheral nerves. But their in vitro isolation, culture and purification are difficult. Acellular nerve allografts (ANA) have a great effect on repairing peripheral nerve defects, and it can induce marrow stromal cells (MSCs) into Schwann-like calls. Accordingly, MSCs can be used as seed calls theoretically in constructing tissue engineered peripheral nerves, instead of Schwann cells OBJECTIVE: To observe the repairing effect of tissue engineered peripheral nerves constructed with MSCs on sciatic nerve defects and to assess the feasibility of repairing peripheral nerve defects with MSCs as seed calls. DESIGN, TIME AND SE'I-rlNG: A randomized control animal experiment was conducted in the laboratory of Basic Medicine Collage of Dali Collage from July to December in 2008.MATERIALS: A total of 30 SD rats were divided randomly into 3 groups, with 10 in each group. In MSCs+ ANA group, end-to-end anastomosis were performed with 10/0 non traumatic suture to broken ends of rat sciatic nerves and tissue engineered nerves cultured using MSCs combined with ANA; In ANA group, ANA were bridged to broken ends of sciatic nerves; In METHODS: The 10ram sciatic nerve defects in rats were repaired using MSCs-constructed tissue engineered peripheral nerves, whose repairing effects were evaluated at week 12 post transplant through sciatic function index (SFI), gastrocnemius wet weight recovery rate, S-100 immunohistochemical staining and electron microscope observation, etc. MAIN OUTCOME MEASURES: Morphological changes of calls were observed during the culture of compounds; SFI and gastrocnemius wet weight recovery rates were detected after transplantation; New myelinization, axon growth and nerve fiber distdbuUon were observed with toluidine blue staining; Schwann calls growth and nerve fiber regeneration were observed with transmission electron microscope combined with S-100 protein immunohistochemical staining method.RESULTS: The detection results showed that SFI and gastrocnemius wet weight recovery rate were higher in the MSCs+ ANA group than in the ANA group (P < 0.05). Compared the ANA group, the MSCs+ ANA group had more S-100 proteins expressed in its compounds obviously, and the number, the diameter of its myelinated nerve fibers as well as its myelin sheath thickness were all greater than those of the ANA group (P < 0.05). The repairing effect of the MSCs+ ANA group was close to that of the autotransplantation group.CONCLUSION: MSCs-constructed tissue engineered nerves have a better effect on repairing peripheral nerve defects than ANA, and MSCs as seed cells have a high value in peripheral nerve tissue engineering.

Puerarin reduces ethanol-induced-apoptosis of spermatogenic cells in rat testis / 基础医学与临床

Objective To investigate potential protection by protect against ethanol-induced apoptosis of spermato-genie cells in rat testis. Methods Thirty SD adult male rats were randomly divided into three groups: normal group, alcohol group and puerarin group. At 40th day, BCL-2 and BAX of spermatogenic cells of testis tissue were checked by RT-PCR and immunohistochemistry; Apoptosis of spermatogenic cells was determined by TUNEL. Re-suits The results of RT-PCR and immunohistochemistry indicated that BCL-2 and BAX of spermatogenie cells were not significanty different between puerarin group and normal group, but there was the significant difference between alcohol group and puerarin group (P <0.01). Apoptosis of spermatogenic cells in alcohol group was significantly higher than normal group. Conclusion Spermatogenic cells could generate apoptosis by changing the expression of BCL-2 and BAX. Puerarin could inhibit this damage of didymus by alcohol.

Puerarin reduces ethanol-induced-apoptosis of spermatogenic cells in rat testis / 基础医学与临床

Objective To investigate potential protection by protect against ethanol-induced apoptosis of spermatogenic cells in rat testis.Methods Thirty SD adult male rats were randomly divided into three groups: normal group,alcohol group and puerarin group.At 40th day,BCL-2 and BAX of spermatogenic cells of testis tissue were checked by RT-PCR and immunohistochemistry;Apoptosis of spermatogenic cells was determined by TUNEL.Results The results of RT-PCR and immunohistochemistry indicated that BCL-2 and BAX of spermatogenic cells were not significanty different between puerarin group and normal group,but there was the significant difference between alcohol group and puerarin group(P