Objective To investigate the effect of nerve growth factor (NGF) on the bone induction of recombinant human bone morphogenetic protein-2 (rhBMP-2) through local application of NGF in the osteoinductive process of BMP. Methods Thirty-six ICR mice were divided into the experimental group and control group at random, and rhBMP-2/collagen composite was implanted into the right thigh muscle pouch of each group. NGF or vehicle was daily injected into the implanted sites of BMP, respectively, for 7 days starting from the third day after surgery. At d10, d20 and d30 after implantation, new bone formation was measured radiographically, biochemically and histologically to compare the osteogenetic capacity of the two groups. Results In both groups, new bone formation was found at d10. However, there was significantly more new bone in the experimental group according to histological and radiographic examinations. At d10 and d20, alkaline phosphatase activity of the local tissue in the experimental group was significantly greater than that in the control group, and calcium and phosphonium contents of samples were also greater in the experimental group. Arrangement of collagen fibers became more regular in the experimental group than that in the control group. Conclusion NGF possesses synergistic effect on ectopic bone formation induced by rhBMP-2.

OBJECTIVE: To explore the correlation between voriconazole's trough concentration and mental abnormity. METHODS: Retrospectively analyze voriconazole's trough concentration and the occurrence of mental abnormity in 151 patients from May 2016 to May 2017 in Xiangya Hospital. RESULTS: In 151 patients, Insomnia rate is 21.85%, binary logistic regression analysis showd that the insomnia rate is positively related to voriconazole's trough concentration(P = 0.02). Illusion rate is 15.23%, binary logistic regression analysis showd that the illusion rate is positively related to voriconazole' s trough concentration (P = 0.002). CONCLUSION: There is a significant correlation between voriconazole's mental abnormity and trough concentration.

Objective To investigate the effect of tamoxifen on pituitary tumor transforming gene ex-pression in C6 cell and tumor growth of glioma in vivo. Methods Animal models were established on 32 SD rats with C6 cells of glioma. The rats bearing with C6 glioma were divided into 4 groups randomly, which were treated without tamoxifen or with different doses of tamoxifen(0. 02 mg · kg-1 · d-1, 0. 2 mg · kg-1 · d-1, 2mg · kg-1 · d-1) once a day for 20 days. The dimension of tumors were measured, the tumor inhibition rates were caculated, and living state of the rats were observed. The expression of PTTG mRNA was detected by RTPCR. Results All kinds of doses of tamoxifen could inhibit the tumors growth in rats with C6 glioma, and the tumor volume were reduced by 47.6%, 35.5% and 21.2% in the high-, middle-and low-dose groups respec-tively, there were significantly differences among the 4 groups ( P < 0. 05 ). Low-, middle- and high- dose of tamoxifen all could inhibit the expression of PTTG mRNA, and there were significantly differences among the 4 groups in PTTG mRNA expression ( P < 0. 05 ). Conclusion Tamoxifen can inhibit PTTG expression and the growth of glioma cell line C6 in vivo significantly in dose-dependent manner, which provides a theory basis for clicinal use of tamoxifen in patients with gliomas.

Objective To investigate the effects of different doses of dexmedetomidine used in SEP and MEP monitoring in patients undergoing neurosurgery. Methods Eighty patients undergoing neurosurgery receiving SEP and MEP monitoring were randomly divided into 4 groups(n = 20 each):group C,group D1,group D2 and group D3. In groups D1 ,D2 and D3 ,dexmedetomidine 0.5 μg/kg was infused over 10 minutes before anesthesia induction,and then was infused at a rate of 0.1,0.3 and 0.5μg/(kg·h)respectively toward the end of operation. Group C received the equal volume of normal saline. HR ,MAP and BIS were recorded at admission to the operating room(T1),skin incision(T2),when the muscle relaxants were stopped(T3)and 50 minutes later(T4). The current intensity and the time when first MEP was induced after muscle relaxant was stopped ,the amplitudes and latencies of SEP(N20-P25,N20)and MEP on thenar muscle at T4,the total consumption of propofol,and development of adverse affects were also recorded. Results Compared with groups C and D1,HR and MAP were decreased at T2-T4 in groups D2 and D3(P<0.05). The amount of propofol consumed were lower in groups D2 and D3 than in groups C and D1(P < 0.05). The current intensity inducing MEP was lower and the amplitude of MEP at T4 was higher in group D2 than in groups C,D1 and D3,and the situation was same in group D3 than in group C (P<0.05). No significant change was found in the other parameters in groups C ,D1 ,D2 and D3(P>0.05). Conclusion Dexmedetomidine infused at 0.3 μg/(kg · h) after infusion of a loading dose of 0.5 μg/kg could improve monitoring quality of MEP through reducing the amount of propofol consumed ,have less inhibition on MEP than other groups,have no obvious effects on SEP,andmaintain hemodynamic stability.

Objective To observe the effect of lnng-term in vitro culture on the biological properties of adipose-derived stem cells(ADSCs)as seeding cells of tissue engineering.Methods The surface makers and apoptosis of primary and passaged human ADSCs were identified by flow cytometric analysis.Osteogenic differentiation of ADSCs at different passages were identified by alkaline phosphatase(ALP),Von Kossa staining and RT-PCR respectively.Results The surface marker expression of mesenchymal stem cells on ADSCs was high and did not change with passages of the cells.The early apoptosis rate of the cells was 1% to 2%,and increased insignificantly from passage one to passage nine.The osteogenic potential of ADSCs confirmed by ALP,Von Kossa staining and RT-PCR was maintained to as late as passage eight.Conclusion Since the biological properties of ADSCs are stable,they can be served as optimal seeding cells for tissue engineering and regenerative research.

The aim of this study was to detect the localization and level of tyrosine phosphorylated proteins during in vitro capacitation of guinea pig sperm. Sperm from mature guinea pigs were incubated in modified TALP under 5% CO_2 in air at 37 ℃. The capacitation effect was assessed by chlortetracycline (CTC) staining. Western blotting and indirect immunofluorescence were used to analyze the level and localization of tyrosine phosphorylation. The results showed that guinea pig sperm underwent a time-dependent increase in protein tyrosine phosphorylation during the in vitro capacitation and the percentage of protein tyrosine phosphorylated sperm increased from 36% to 92% from the beginning of incubation to 7h incubation. Also, there was a shift in the site of phosphotyrosine-specific fluorescence from the head of sperm to both the head and the flagellum of sperm. Moreover, there were three proteins phosphorylated in this experiment. After 0 to 0.5h incubation, the protein of 40kDa was detected by anti-phosphotyrosine monoclonal antibody, and the intensity of this protein increased in the following incubation. Then, after 1h incubation, another protein of 80kDa was found and the level of this protein reached the highest point at 3h. Also, in 3h incubation, a protein of 45kDa was detected and the intensity of this protein increased in the following incubation.

The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.

OBJECTIVE:To improve the drug delivery procedures and reduce the defects occurred in drug delivery.METHODS:The five procedures of six sigma including defining,measuring,analyzing,improving and controlling were applied to improve the drug delivery in our hospital's single dose dispensing(UDD)center.RESULTS:Due to the application of the six sigma method,the defect rate of drug delivery dropped from 25.1%(before the initiation of the six sigma method)to 5%(after the initiation of the six sigma method),which was significant lower as compared before the initiation of the six sigma method.CONCLUSION:Six sigma method contributed to the reduction of defect rate of drug delivery,so it deserves to be widely used in each management link in UDD center so as to improve the quality of dispensing.

Objective To investigate the relationship of acute graft versus host disease (aGVHD) and the regulatory T (Treg) cells,NK cells as well as their function-related cytokines such as IL-10,TGF-β,and perforin in mice with allogeneic bone marrow transplantation.Methods H-2 completely mismatched C57BL/6→ BALB/c aGVHD mice model was constructed.Flow cytometry analysis was used to detect the proportion of CD4+CD25 + Treg cells and NK-1.1+ NK cells in the spleen of aGVHD mice after transplantation.ELISA method was used to detect the serum levels of IL-10,TGF-β and perforin in the aGVHD mice intervened with CSA prophylactic or not.The normal C57BL/6 mice were used as controls.Results Compared with those of normal mice,the proportion of Treg cells in aGVHD mice after transplantation was decreased (mean 3.6％ vs.1.55％) and the proportion of NK cells increased (mean 3.3％ vs.11.5％).In the aGVHD mice treated with cyclosporine A,serum IL-10 expression level was significantly increased (treated group (125.79 ± 0.27)pg/mL,untreated group [(103.09 ± 3.27)pg/mL,P＜0.01)],TGF-β expression level was increased [(252.05 ±7.84)pg/mL vs.(241.61±15.41)pg/mL,P＞0.05],perforin expression level was significantly increased [(186.97 ± 4.68)pg/mL vs.(144.35 ± 14.42)pg/mL,P＜0.01].Conclusion ① The occurrence of aGVHD is correlated with the decreased number of Tree cells after transplantation in mice.Treg cell function-related cytokines IL-10 and TGF-β are involved in the treatment of aGVHD by cyclosporine A-mediated immunosuppression.②NK cells are involved in the occurrence of aGVHD after allogeneic bone marrow transplantation,and the increased level of perforin is related to the inhibition of aGVHD.

AIM:To construct the recombinant eukaryotic expression plasmid pEGFP-C1-SR-A I for the high expression in 293T cells in order to identify functions of savenger receptor-A I(SR-A).METHODS:The primer was designed according to MSR1 cDNA and pEGFP-C1-SR-A I was constructed by standard molecular cloning technique and enzyme digestion.After sequencing,the plasmid was transfected into 293T cells by lipidosome method.The expression of scavenger receptor-A I was identified by RT-PCR and Western blotting.The foam cells were evaluated by the formation of lipid granules in the cells with oil red staining.Cell adhesion was analyzed by cell adhesion assay.RESULTS:24 h after transfection,SR-A I mRNA was highly expressed and the high level of the protein was detected.The ratio of foam cell formation was doubled,the efficacy of cell adhesion was enhanced two times compared to the control group and the empty vector group.CONCLUSION:The recombinant eukaryotic expression plasmid has been constructed successfully with enhancing the function of uptake ox-LDL and adhesion in 293T cells by overexpression of SR-A I.