Objective: To explore the role of anti-?? T cell receptor(TCR) and anti-CD80 monoclonal antibodies(mAbs) combined with donor bone marrow cells(BMCs) infusion in the induction of murine skin allografts tolerance.Methods: On day 0,2?10~8 BMCs of BALB/c mice were injected into recipient C57BL/6 mice via the tail vein,meanwhile,an intraperitoneal injection of TCR?? mAb(500 ?g) was given.On day 2,CD80 mAb was administered intraperitoneally.Skin grafting was performed on day 6.Delayed type hypersensitivity(DTH),mixed lymphocyte reaction(MLR),IL-2 reverse assay of MLR,adoptive transfer assay and chimerism detection were performed at different time points and tolerance mechanisms were investigated.Results: The mean survival time(MST) of BALB/c skin allografts in C57BL/6 recipients that were treated by anti-TCR?? and anti-CD80 mAbs combined with donor BMCs infusion was 70 days.DTH and MLR assay indicated that the tolerant mice displayed significant hyporesponsiveness.The result of IL-2 reverse test showed that clone anergy was probably involved in the formation of tolerance in the tolerant C57BL/6 mice.In vivo and in vitro adoptive transfer assay,suppressive activity in the spleens of tolerant C57BL/6 mice was observed.Chimerism existed in both the thymus and spleen of the tolerant C57BL/6 mice.The chimerism level gradually declined with time.Conclusion: Treatment of anti-TCR?? and anti-CD80 mAbs combined with donor BMCs infusion can successfully induce a long-term tolerance in BALB/c mice to C57BL/6 skin graft.Multiple mechanisms,including clone anergy,suppressor cells and chimerism are involved in the tolerance.

Objective To study the effect of Danggui Buxue Tang (DBT) on immunological reconstitution of bone marrow transplantation (BMT) in mice. Methods BALB/c mice were irradiated by 137Cs γ once 8.5 Gy, then the mice were engrafted with bone marrow cells (107 cells/mouse) within 4 hours lethal irradiation. And the mice were fed by DBT every day for 15 days. Flow cytometry technique combined with immunological methods were performed to evaluate immunological reconstitution of BMT mice in 30 and 60 days pest-transplantation. Peripheral blood RBC and WBC were counted, and nucleated cells were assayed in recipient bone marrow. Lymphocyte numbers in thymus and periphery were counted and subpopulations of the two origins were observed respectively. Lymphocyte proliferation induced by Con A and LPS, plaque-forming cell (PFC), delayed type hypersensitivity (DTH) were also examined in 30,60 days in post-transplantation respectively. Results Compared with BMT mice, BMT mice treated with a certain dose of DBT could increase the number of peripheral blood RBC and WBC in the recipients, and also could increase that of nucleated cells significantly. BMT mice treated with DBT could increase lymphocyte numbers in thymus and periphery, and improve thymocyte subpopulations, resulting in enhancement in immune function. Conclusion DBT can enhance the immunological reconstitution of BMT mice.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; diagnosis ; surgery ; Lumbar Vertebrae ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Thoracic Vertebrae ; surgery ; Treatment Outcome

RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.

Objective To establish and evaluate a gene microarray for determination katG mutations of Mycobacterium tuberculosis isolates associated with resistance to isoniazid(INH).Methods A panel of probes were designed and gene chips were prepared by dotting.Mycobacterium tuberculosis isolates resistance to 5 drugs was determined by proportional dilution methods.Amplicons of Mycobacterium tuberculosis isolates were detected by our chip and sequenced.Results The drug resistance rate of the isolates to at least one of the anti-tuberculosis drugs was 70.8%(97/137).45 strains out 137 Mycobacterium tuberculosis isolates was resistant to INH(32.8%).katG was successfully amplified from 100% of the susceptible strains and 88.9%(40/45)resistant strains.4 of 45 INH resistant isolates' katG were deleted.27 of 40(67.5%) katG has been detected to have katG 315 codon mutations.The mutations were 315 AAC(Asn,13/40), ACC(Thr,6/40),ACA(Thr,4/40),ATC(Ile,2/40),AGC(Arg,2/40).The mutation rate of katG analyzed by gene chips we prepared were identical to katG sequencing.Conclusion The gene microarray techniques we developed for determination of Mycobacterium tuberculosis resistance to INH are specific, sensitive and may be used as an alternative in clinical laboratory.

OBJECTIVE: In recent years, available evidence from basic and clinical research on Alzheimer disease(AD) suggests that oxidation stress is involved in the occurrence and development of AD, and that antioxidant treatment can improve the intelligence of patients with AD and delay age-dependant cognitive dysfunction. Although results of basic and clinical research on the therapeutic effects of antioxidants on AD are inconsistent, a large number of available data suggest that these studies are of significance. Basic pharmacological studies on natural antioxidant TA99 series indicate that they are promising novel drugs for AD. Thereby, this study made a review of their experimental basis in the treatment of AD and existing problems.DATA SOURCES: Related articles published between January 1991 and December 2004 were searched by the computer in Medline database with such key words as Alzheimer disease, antioxidant, Ginkgo biloba extract, TA9901,acetylcholine, and senescence-accelerated mouse in different combinations and with the language limited to English. Meanwhile, related articles were alsosearched in CDMA \Wanfang database with the same key words in Chinese.STUDY SELECTION: Literature involving intervention group and control group were screened in the first trial, and then non-randomized trials were excluded and the rest were searched for the full text.DATA EXTRACTION: Of the 24 basic and clinical randomized and non-randomized trials on antioxidants in the treatment of AD collected, 17 accorded with the inclusion criteria and the other 7 were excluded.DATA SYNTHESIS: Intervention in the 17 trials emphasized the pathogenesis of AD from amyloid β proterin(Aβ) synthesis, gathering to senile plaque formation, and the enhancement of Aβ gathering and neuronal apoptosis by peroxidative injuries of free radicals. Both in vitro and in vivo studies were conducted: the effect of Aβ on neurons of different regions was observed with cell culture; transmission electromicroscope and sulfrin T (Th-T) fluorescence assay, Fuliye-transform infrared(FT-IR) spectrum apparatus, electron magnetic resonance(EPR), and round spectrum were used to detect the inhibitory effect of TA99 series on Aβ gathering and fibroplasia in vitro, as well as the influence on Aβ gathering in vivo. Senescence accelerated mouse (SAM) -P/8 was adopted to establish AD model and behavioral studies such as Morris water maze were used to investigate their effect on learning and memory. Meanwhile, the clearance of intracerebral amyloid granular deposition due to TA99 was also observed with hexamic argent staining. The effects of TA series on Aβ target and possible mechanism were fully revealed, and basic pre-clinical data collection was almost completed.CONCLUSION: TA9901 plant extractions have been proved to inhibit Aβ gathering and fibrosis, and improve learning and memory of SAM-P/8 rats. Moreover, TA9902 prepared by TA9901 combined with EGb761, another synergic herb, has an obvious anti-neurotoxic effect by inhibiting Aβ gathering, fibrosis and secondary structural changes. Further pharmacological research is needed and will have a promising prospect.

Although previous reports showed drug-eluting stent (DES) could effectively inhibit neointima formation, in-stent restenosis (ISR) remains an important obstacle. The purpose of this study was to investigate different effects of paclitaxel on proliferation and cell cycle regulators between vascular smooth muscle cells (VSMCs) and vascular endothelial cells (VECs) of rats in vitro. The cultured VSMCs and VECs of rats from the same tissues were examined by using immunohistochemistry, flow cytometry and Western blotting in control and paclitaxel-treated groups. The results showed paclitaxel could effectively inhibit proliferation of VSMCs and VECs. However, as compared with VECs, proliferation of VSMCs in paclitaxel-treated group decreased less rapidly. The percentage of cells in G0-G1 and G2-M phases was reduced, and that in S phase increased after treatment for 72 h. The expression of cyclin D1 and B1, p27 and PCNA in VSMCs of paclitaxel-treated group was up-regulated, but that of p21 down-regulated as compared with VECs. It is concluded that there are significant differences in the expression of cell cycle regulators and proliferation rate between paclitaxel-treated VSMCs and paclitaxel-treated VECs, suggesting that the G1-S checkpoint regulated by paclitaxel may play a critical role in the development of complications of DES, which provides new strategies for treatments of ISR.

Objective:To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein.Methods:A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured;the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry.Results:Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P<0.05 or P<0.01);the indexes metioned above of the mice in MUC1-MBP+ R848 were higher than those in MUC1-MBP+BCG group;the IL-4 levels had no significant difference(P>0.05).Compared with control group,the proprotions of CD3+T,CD4+T,and CD8+ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01).Conclusion:Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.

CBS(Case Based Study)teaching method is given in a case requiring students to answer a series of questions surrounding the case.In the teaching process,students of small groups are required to find their own solutions.Compared with tradition method,this mothed can improve students'academic performance(P

Aim To evaluate the inhibitive and induc-tive effects of Polygonum capitatum water extract on main cytochrome P450 isoforms in human and liver mi-crosomes of mouse in vitro for predicting the herb-drug interactions in clinical application. Methods The in vitro inhibitory effect was evaluated by incubating Po-lygonum capitatum water extract with the probe sub-strates of main phase I metabolic enzymes in human liver microsomes, including CYP1A2, CYP2E1, CYP2C9,CYP2C19 and CYP3A4. Mice were adminis-tered with Polygonum capitatum water extract at dosage of 0 . 58 g · kg-1 and 1 . 16 g · kg-1 by gastric lavage for successive 7 days and 14 days, then the cocktail-LC-MS/MS method was applied to assess the inductive effect of main CYP450 isoforms in mouse liver micro-somes. Results The IC50 values of Polygonum capita-tum water extract on main CYP450 isoforms in human liver microsomes were from 849 . 6 mg · L-1 to 2 287 mg·L-1 . Compared with the blank control group, the activites of CYP2C9 and CYP3A4 in 1. 16 g·kg-1 7 d group were about 49 . 9 % and 21. 1 % higher ( P <0. 01, P < 0. 05 ) respectively, the activities of CYP2C9 and CYP3A4 in 0. 58 g·kg-1 7 d group were 27. 6 % and 15. 5 % higher ( P <0. 01 , P <0. 05 ) respectively, the activities of CYP2C9 and CYP3A4 in 1. 16 g·kg-1 14 d group were 67. 5 % and 32. 1 %higher (P<0. 01) respectively, while the activities of CYP1 A2 , CYP2 E1 and CYP2 C19 were not increased significantly in Polygonum capitatum treatment group. Conclusions Polygonum capitatum water extract do not show the inhibitory effect on main CYP450 in hu-man liver microsomes. There is induction on CYP2C9 and CYP3 A4 in mouse liver microsomes by Polygonum capitatum water extract.