Objective:To prepare the recombinant neuron protective protein TAT-Bcl-XL in E.coli expression system and to determine its anti-apoptosis activity.Methods:DNA fragment of TAT-Bcl-XL was obtained by RT-PCR using primers that were specific for Bcl-XL gene. Prokaryotic expression vector(pTBTOPO) was constructed by insertion of TAT-Bcl-XL DNA fragment into pCRT7/CT-TOPO vectors, and the recombinant protein was detected by SDS-PAGE and Western blot analysis using anti-V5 tag epitope antibody as the primary antibody. The recombinant fusion protein was purified by affinity chromatography, and the anti-apoptosis function of the purified protein was determined by flow cytometry.Results:A molecular weight of 30 000 protein was detectable in both SDS-PAGE and Western blot analysis. Immunofluorescent staining showed the fusion protein was distributed in cell plasma after incubation with 200 nmol/L fusion protein. Flow cytometry data indicated that the recombinant protein could enhance cell survival by 40% when 293T cells were treated with 250 ?mol/L zinc chloride as a supplementation in cell culture media.Conclusion:High level expression of the reported neuron protective protein was obtained in E.coli expression system, and the purified recombinant proteins remained its anti-apoptosis activity in our preliminary characterization.

Objective To observe the rat tracheal stem cells in situ.Methods Tracheal rings of Wistar rats treated with 5-FU to make an injure model in vitro then observing the process of regeneration in sequence time from 3*!hours to 48*!hours. Use light microscope to detect the development of tracheal epithelium,assay the level of proliferating cell nuclear antigen (PCNA) immunohistochemistry and perform Hoechst33342 staining to search the negative staining cells. Results Twelve hours after 5-FU it could be seen that tracheal mucosa exfoliated with nuke-nucleus like cells nailed just above the basement membrane,6*!hours after removing 5-FU we could see that the tracheal rings were covered with squamous epithelium. It could be seen that PCNA immunohistochemistry negative staining cells located bordering on the positive one. Meanwhile under fluorescent contrast microscope negative cells with no Hoechst dye combinations were among most of the positive cells. 12*!hours after removing 5-FU mucous granular cells and cilia cells could be seen. After 48*!hours the whole tracheal rings were covered with pseudostratified columnar ciliated epithelium.Conclusion 5-FU is a cell-cycle specific antimetabolite cytotoxicant.It can make the cycling cells degeneration and necrosis but has little effect on the G 0 cells. Few G 0 cells which look like nuke nucleus cells randomly on the basement have the ability to efflux the dye Hoechst33342.The tracheal rings are regenerated through these cells differentiation and proliferation.

Iron overload is a metabolic disorder characterized by excessive iron deposition in the liver,pancreas,heart,endocrine organs, etc.,resulting in structural damage and dysfunction.The liver is the primary organ for iron storage,and excessive iron deposition induces liver inflammation and fibrosis,which may progress to cirrhosis and even liver cancer,with a poor prognosis.Accurate evaluation and effec-tive treatment can reduce liver injury caused by iron overload and improve patients′survival.

Objective To evaluate the application of diffusion-weighted MR imaging in hepatic echinococcosis.Methods Diffusion-weighted MR imaging and measurement of apparent diffusion coefficient (ADC)were performed in 23 patients with 30 hepatic echinococcosis(cystic echinococcosis 24, alveolaris echinococcosis 6).Results The mean ADC values of cystic echinococcosis, alveolaris echinococcosis and normal liver were (3.50?0.35)?10 -3 mm2/s,(2.22?0.22)?10 -3 mm2/s and (1.41?0.09)?10 -3 mm2/s respectively.The difference between the mean ADC values was significant(?

Aged ; Amiodarone ; adverse effects ; Chemical and Drug Induced Liver Injury ; etiology ; Drug-Related Side Effects and Adverse Reactions ; Humans ; Male

[Objective] To develop new bate titanium alloys made of titanium,niobium and zirconium(Ti-Nb-Zr)which have higher strength,lower moduli of elasticity and better biocompatibility for orthopedic implants.[Methods]Alloy elements including titanium,niobium and zirconium were melted together and forged into forging Ti-Nb-Zr.Four kinds of alloys were developed because of the different proportion of alloy elements.All alloys were studied using X-ray diffraction analysis,biomechanical measurements,and scanning and transmission electron microscopy.[Results]X-ray diffraction analyses showed that all the candidates were beta alloys.The as-cast alloys had the strength values in 584~718 Mpa,elastic modulus values in 59~68 Gpa.The peak-aged alloys had the strength values in 706~874 Mpa,elastic modulus values in 59~66 Gpa,elongation rates in 5.4%~20%.The forging procedure could increase the strength significantly but almost have no influence in elastic modulus of Ti-Nb-Zr.[Conclusion]The Ti-Nb-Zr beta titanium alloys developed in our lab have lower modulus and high strength both.It is a promising material for orthopedic implants.

[Objective]To evaluate the biocompatibility of a new bate titanium alloys made of titanium, niobium and zirconium (Ti-Nb-Zr) which have higher strength and lower elastic modulus by in vitro study.[Method](1)the cell toxicity of Ti-Nb-Zr was evaluated using L-929 cell line's relative growth rate (RGR) which was transformed into 6 toxicity ranks in according with the national standard;(2)the J774A.1macrophages cultured with Ti-Nb-Zr particles whose diameter was about 1 ?m for 24-48 hours were observed using inverted phase contrast microscopy and transmission electron microscopy. Expression of IL-6 and TNF-? mRNA in these cells were measured using RT-PCR technique. The TNF-? in the supernatant was also measured using ELISA method. All the results were compared with the traditional orthopedic implants metal materials such as Ti-6Al-4V and Co-Gr-Mo.[Result]The cell toxicity of Ti-Nb-Zr was ranked zero degree. The particles could be seen in the macrophages under the light microscopy. Ti-Nb-Zr particles made the J774.A1 macrophages less configurative changes than Ti-6Al-4V and Co-Gr-Mo did.Expressions of IL-6 and TNF-? mRNA in J774.A1 cell were increased after stimulation for 48 hours by all kinds of metallic materials. The TNF-? in the supernatants of Ti-Nb-Zr group were less than that of the Ti-6Al-4V and Co-Gr-Mo groups (P

[Objective]To develop new beta titanium alloys made of titanium, niobium and zirconium (Ti-Nb-Zr) which have higher strength, lower moduli of elasticity. Its biocompatibility in vivo is evaluated in this paper. [Method](1) 24 SD rats were used in the modified air pouch model experiments. Air was injected subcutaneously on the back of rats to form the pouches and the Ti-Nb-Zr particles were introduced into them. The liquid in the pouches was harvested 48 hours later for IL-6 and TNF-?ELISA procedure and the capsules for histology examination. All the results were compared with Ti-6Al-4V. (2) Ti-Nb-Zr bone plates were fixed onto tibias of rabbits. The capsules around the plates and the bone under them were observed at 4, 8, 12, 24 and 36 weeks respectively. Ti-Nb-Zr-bone interfaces were emphasized in this procedure. This time the control material was stainless steel. [Result](1)Ti-Nb-Zr and Ti-6Al-4V particles made the air pouches secret more TNF-? than that of the control (P

To elucidate the processing mechanism of Herba Epimedii from the new view of the intestinal absorption and metabolism.From analyzing the literatures of processing mechanism and togethering with the research of our lab,a new idea of processing mechanism of Herba Epimedii was brought:the pharmacologically active and easier absorbed flavonoids might be present more in the herbs when changing the heating processing parameters and thereby increased or maintained the efficacy.This thesis first pointed the new idea and method that the intestinal absorption and metabolism of herbs should be considered when studying the mechanism of processing.

Objective To investigate the effect of artorvastatin on macrophage accumulation in tubulointerstitium of unilateral ureteral obstructive (UUO) nephropathy and its possible mechanism.Methods Twenty-one rats were randomly divided into three groups: sham-operatipn, UUO, UUO+ artorvastatin. Immunohistochemistry staining of CD68 and M-CSF was used to define the macrophage accumulation and expression of interstitial M-CSF. Lipid profile in these groups was also determined. Results CD68 + cells and M-CSF expression were significantly increased at day 10 after UUO operation, this kind of CD68 + cell accumulation and M-CSF expression up-regulation were ameliorated by artorvastatin treatment. In UUO and atorvastatin treated groups, the number of macrophage was positively correlated with tubulointerstitial M-CSF expression. There was no significant difference about serum lipid among the three groups. Conclusion Atorvastatin can reduce interstitial macrophage accumulation in UUO nephropathy. This therapeutic effect might relate to down-regulation of tubulointerstitial M-CSF expression.