The mouse heart was transplanted into the neck of rat using the cuff technique deseribed by Heron with a little modification.The graft accending aorta to the host common carotiel artery and the graft pulmonary artery to the host exteral jugular vein were performed.22 times of formal transplantation were done with the successful rate being 86%.The mean survival time was 2.10±0.80 days.The method is easy,quick and reliable.The superficial cervical localigation of the transplant is easily observed.This is a good model to observe the hyperacute rejection(HAR),to induce the translantation tolerance,and to screen the new immunosuppressive agents.

Wistar-to-SD rat pancreas transplantation was performed in 35 SD rat recipients with diabetes-induced.Donor antigen was administered through IV or PV and the antigen treatment was performed before or during the transplantation.Graft vein was drained into IV or PV.The results showed that donor antigen pretreatment via PV combined with graft drainage PV synergistically prolongs the pancreas graft survival,and the effect of simultananeous antigen treat-ment during transplantation is not as satisfactory as that of antigen pretreatment,but may be more practical.

Objective To investigate value and effect of the bronchus was cut out and bronchoplasty for lung cancer. Methods Twenty-one patients with lung cancer which all invased out one bronchial orifice and precluded standard lobectomy. The present of endobronchial tumor was cut out according to the lesion region during operation, the bronchoplasty performed by health bronchial wall. Results There were squamous carcinoma in 15 patients; adenocarcinoma in 4; small cell lung cancer and carcinoid each one patient. Staging: stage Ⅰ in 3 patients; stage Ⅱ in 12, stage Ⅲa in 6. The 19 patients were uneventful after operation. The 2 cases died from acute pulmonary embolism or gastric irritable ulcer with large bleeding in perioperation. Respiratory failure in all patients had not occurred. Bronchoscopy performed immediately after operation in 16 patients and in the period of following up 6-36 months in 12 patients, the mucosa of bronchoplastic sites was smooth, no bronchial stenosis and no tumoral relapse.The period of following-up was 4-72 months, median survival 42 months after operation. Five patients died from the metastases or relapse. Life time in survival patients was well. Conclusion The bronchial resection by cut out and bronchoplasty for lung cancer had a good results in near future and long period, and proved individualized and adapted selective type of resection for bronchoplasty of lung cancer.

Objective To investigate the results of dynamic hip screw (DHS) intemal fixation for the treatment of femoral intertrochanteric fractures. Methods Thirty patients with femoral intertroehanteric fractures treated with DHS internal fixation were enrolled in the study. The classification of fracture, operative methods, postoperative treatment and therapeutic results were assessed. The mean follow-up period was one year. Results All the patients completed the follow-up. Fractures healed up without coxa vara, external rotation or shortening deformity of the lower extremity in all patients. Good hip joint function was obtained in 86.7% of the patients. Conclusion DHS internal fixation is an ideal method for the treatment of femoral intertrochanteric fractures.

BACKGROUND: Macrophage colony-stimulating factor (M-CSF)/receptor activator of nudeer kappa B ligand (RANKL), two types of cytokines co-induce myeloid stem cells to form osteoclasts, is a kind of new method to harvest ostaoclasts with high purity and quantity, but there is lack of uniform cultivation standard. OBJECTIVE: To construct an effective M-CSF/RANKL induced mice myeloid stem cells inducing osteoclast differentiation cultivation system. METHODS: Myeloid stem cells ware obtained from ICR mice and then cultured for 24 hours in a-minimum essential medium containing M-CSF, at cell density of 10~7/L, 10~8/L, 10~9/L. Then 10 μg/L M-CSF and 20, 50, 100 μg/L RANKL were added into culture medium. Tartaric-resistant acid phosphatase stained was performed to observe the transition process from stem cell to osteoclast, as well as cell morphology and stain situation after culture, and positive stained osteoclasts were counted. We compared the influence of different induction conditions to the quantity of osteoclast. RESULTS AND CONCLUSION: A small quantity of osteoclasts contained many red positive beads in the intracytoplasm were observed at 3 days. There were positive beads with hypochromatic dikeryon in cells. A large amount of positively stained osteoclests were seen after 6-day cultudng, which maintained dikaryon. After 9-day culturing, positively stained colossal multinudear cells occurred, became larger and maintained three nuclei. At certain cell density, 100 μg/L RANKL could induce to form more osteoclasts compared with other 2 concentrations (P < 0.05); at certain RANKL concentration, the osteoclasts formation at cells density of 10~8/L was dramatically greater than other 2 cell densities (P < 0.05); the number of osteoclasts was the most when the concentration of RANKL was 100 μg/L and cell density of 10~8/L (P < 0.05). When osteoclasts are induced by M-CSF/RANKL from mudne myeloid stem cells, the best concentration of RANKL is 100 μg/L and cells density is 10~8/L.

Objective To establish the quality control standard for Yi-Li-Kang tablets. Methods Astragalus membranaceus and Herba Epimedh were identified by TLC. The content of Icariin was determined by HPLC. Results The linear range of Icariin was 0.288～2.016 μg. The average recovery was 100.83%, and RSD was 1.82 (n=5). Conclusion The method is simple, accurate and reliable, which can be used to control the quality of Yi-Li-Kang tablets.

Objective To investigate the mechanism of therapeutic action of dexamethasone on asthmatic mice by detecting the levels of IL-25 and IFN-γ in bronchoalveolar lavage fluid (BALF). Methods Balb/c mice with SPF grade were randomly divided into normal control group, asthma group and dexamethasone group. Asthma group and dexamethasone group were sensitized and challenged with ovalbumin ( OVA) . Dexamethasone group was intraperitoneally injected with dexamethasone one hour before challenging. The mice were executed 24 hours after the last challenge, and the HE stained pathological sections of the right lung were made. Pathological sections of lung were observed. BALF in the left lung was also collected. The total white blood cell count and absolute eosinophile ( EOS) count were observed, and the percentage of EOS was calculated. The levels of IL-25 and IFN-γwere measured with ELISA, and correlation analyses were made. Results The counts of total white blood cell and EOS, and the percentage of EOS were significantly higher in the asthma group than in the normal control group and dexamethasone group (P<0. 05). No differences were found between the normal control group and dexamethasone group. The IL-25 level was higher in the asthma group than in the normal control group and dexamethasone group (P<0. 05), and its level in the dexamethasone group was also higher than that in the normal control group. The IFN-γlevel was lower in the asthma group than in the normal control group and dexamethasone group (P<0. 05), while there was no significant difference between the normal control group and dexamethasone group. IL-25 was negatively correlated with IFN-γin each group. Conclusion Part of the mechanisms of dexamethasone acting on asthma are related to its inhibition on the pulmonary inflammation and promotion on the expression of IFN-γ, and possible inhibition of IL-25 expression.

Objective To identify the mechanisms of oral administered rapamycin on the prevention of restenosis after stent implantation with intravascular ultrasound (IVUS) and pathology.Methods Twenty-four New Zealand white rabbits underwent balloon-induced abdominal aortic wall injury and were given a diet of 1 ％ cholesterol for 8 weeks.Then,the rabbits were divided into three groups:the bare metal stent group(BMS group),the bare metal stent with oral rapamycin group(BMS + RAPA group) and the rapamycin eluting stent group(DES group).Rabbits in the RAPA and BMS + RAPA groups received a daily oral dose of rapamycin(0.5 mg/kg),whereas rabbits in the DES group received no drugs.All the rabbits were euthanized after the 4-week intervention.Serum lipids were measured.IVUS and pathologic studies were performed.The minimal luminal diameter (MLD),external elastic membrane (EEM) area,lumen area (LA),and plaque area(PA),plaque burden(PB) were measured.Mammalian target of rapamycin(mTOR) expression level was examined by immunohistochemistry.Results After the 4-week intervention,there was no significant difference of serum lipid levels among the three groups.IVUS showed that oral administration of rapamycin in the BMS + RAPA groups showed similar effects in reducing PA and PB as the DES group,which all were better than the BMS group.The BMS + RAPA and DES groups showed much more MLD and less lumen reduction,compared with the BMS group( P ＜0.05).Level of mTOR expression of the BMS + RAPA group and DES group was significantly lower than that of BMS group.Conclusions Oral administration of rapamycin demonstrates the same effect in the reduction of plaque burden and stent restenosis as the rapamycin eluting stent.Inhibition of mTOR by rapamycin involves in the stent restenosis.

BACKGROUND:It is reported that tailless-like protein (TLX) plays critical roles in the regulation of early developmental processes in vertebrates, and it plays a key role in stem cells proliferation and differentiation into neurons. OBJECTIVE: To construct recombinant plasmid pEGFPN1-TLX and study the transfection into dermal multipotential stem cells. DESIGN, TIME AND SETTING: Cytogene experiment was performed at the Department of Pathogen Biology, School of Basic Medical Science, the Third Military Medical University of Chinese PLA from March to December 2007. MATERIALS: An adult SD was obtained from the Experimental Animal Center of the Third Military Medical University of Chinese PLA; dermal moltipotential stem cells (DMSCs) were cultured by the Institute of Combined Injury of the Third Military Medical University of Chinese PLA; pEGFPN1 and DH5α was gifted by professor Xu.METHODS: Total RNA was extracted from rat brain tissue to amplify TLX-coded cDNA sequence using RT-PCR. T/A was cloned on pMD18-T vector and determined using BamHI and Hindlll. The products were positive recombinant plasmid pMD18-T-TLX segments, which were sub-cloned in pEGFPN1 to construct recombinant plasmid pEGFPN1-TLX. Finally, pEGFPN1-TLX was transfected into DMSCs.MAIN OUTCOME MEASURES: The fluorescence protein expression was observed under fluorescence microscope at 24 hours after transfection; TLX mRNA expression was detected using RT-PCR; neuronal differentiation was observed using immunohistochemical staining.RESULTS: TLX full length cDNA was successfully cloned into pEGFPN1, and pEGFPN1-TLX was successfully constructed by means of sequence analysis and enzyme cutting identification. As compared with non-transfected DMSCs, pEGFPN1-TLX transfected DMSCs were observed after 10 days, formed resistant clones after 15 days, and shown a green fluorescent protein expression. However, non-transfected DMSCs died at day 10. RT-PCR indicated that pEGFPN1-TLX transfected DMSCs could express TLX mRNA. At day 3 after induction, NF200 positive cells were increased, but glial fibrillary acidic protein positive cells were decreased after induction of pEGFPN1-TLX transfected DMSCs.CONCLUSION: TLX was successfully constructed and transfected into DMSCs. After transfection, neuronal differentiation of DMSCs was enhanced, and the differentiation to gliocytes was inhibited.

Objective To prepare a thermosensitive in situ gel system containing berberine hydrochloride for ocular use. Methods The in situ thermosensitive gel system of berberine hydrochloride was prepared by Poloxamer 407 (P407) and Poloxamer 188 (P188) as thermosensitive materials,the formulation was optimized by determining solution-gelation conversion temperature by using stirring method. The content of berberine hydrochloride was determined by HPLC. Results The gelation temperature of in situ thermosensitive gel containing berberine hydrochloride formulations lowered as the P407 concentration increased,as the P188 concentration increased gradually the gelation temperature initially increased to maximum and then decreased. The gelation temperature all increased after simulated tear fluid (STF) dilution. The fitted equation was established for the gelation temperature with the concentration of Poloxamer solutions after diluted by STF. An optimized formulation by Design-Expert software was freely flowing liquid at 29.7 ℃ and convert to a firm gel at 34.5 ℃ after STF diluted. Conclusion In situ thermosensitve gel system complies with the requirement for ophthalmic application and shows great potential in ocular application.