The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank. The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digestion by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a(+)to obtain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell with IPTG induction. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, the indirect ELISA assay for the detection of rabies virus antibodies in canine serum was applied after management of the optional working condition.

Adult ; Female ; Heart Neoplasms ; Humans ; Pericardium ; Pheochromocytoma

Benzofurans ; analysis ; chemistry ; Dibenzofurans, Polychlorinated ; Dioxins ; analysis ; chemistry ; Humans ; Occupational Exposure ; prevention & control

Objective To explore the causes and the consequences of iatrogenic bile duct injury as well as experiences in its diagnosis and treatment. Method The clinical data of 28 in-patients who suffered from bile duct injury followed by cholecystectomy during January 1995 to September 2011 were collected and retrospectively analyzed. Result Bile duct injuries were found and diagnosed in al 28 in-patients during the processes of reoperation. Different treatments were carried out. 8 cases with mild injury were treated by placing the abdominal cavity drainage. 6 cases with bile duct split were repaired and set up with T tube drainage. 4 cases were initially treated by bile-intestinal Roux-y anastomosis. 1 case received repair operation of bile-intestinal Roux-y anastomosis after the external drainage of bile. 9 cases were initially treated with end-end bile duct anastomosis and 7 of 9 cases with concurrent anastomotic stenosis were performed repairing re-operations twice or more than twice. Among 7 cases, 1 case was performed with the operation of umbilical vein flap repair while 6 cases were treated with bile-intestinal Roux-y anastomosis repair. All cases in this observation were followed up to two years after repairing surgery:2 patients died and the rest 26 patients recovered after treatment. Conclusions The initial repairing operation is critical for iatrogenic bile duct injury and bile-intestinal Roux-y anastomosis should be the first choice of therapy,and it should be operated by experienced surgeons.

Objective To prepare nerve growth factor gradient release system,and explore the promotion of the clinical effects of peripheral nerve regeneration.Methods All 57 cases with peripheral nerve injuries were treated with emergency nerve repair from July 2010 to August 2012,a nerve repair,with meanwhile randomly were divided into group A (19 cases),group B (19 cases) and group C (19 cases).The NGF was used to partial release by using the homemade gradient release system in group A,the same amount of NGF was partial injeced to Gelatin sponge surrounding peripheral nerve in group B.The NGF wasn't injected in group C.All patients respectively underwent neurophysiological examination after 4 weeks and 8 weeks.Comparising with the somatosensory evoked potentials (SEP) and muscle action potential (MAP) between different groups,meanwhile analyzing nerve conduction velocity (NCV).The sensory and motor function evaluation score of BMRC were performed after 24 weeks.Results The peak latency of the SEP after 4 weeks in A,B and C groups were (34.80 ± 3.45) ms,(42.85 ± 2.58) ms,and (51.05 ±3.652) ms,respectively; the volatility were (10.673 ± 2.35) μV,(6.30 ± 1.22) μV,and (4.10 ±0.83) μV,the peak latency of the MAP after 4 weeks in A,B,C groups were(5.29 ±0.82) ms,(7.63 ± 1.35)ms,and (9.98 ± 1.15) ms,the volatility were (485.75 ±65.83) μV,(262.510 ±59.90) μV,and (185.64 ±38.35) μV,the NCV were (25.50 ±3.65) m/s,(19.80±2.35) m/s,and (15.50 ±2.61) m/s.The peak latency of the SEP after 8 weeks in A,B,C groups were (24.68 ±4.50) ms,(31.08 ±4.24) ms,and (39.25 ±4.83) ms,the volatility were (21.22 ± 4.63) μV,(13.68 ± 4.14) μV,and (8.90 ± 1.25) μV.The peak latency of the MAP after 8 weeks in A,B,C groups were (4.71 ±0.35) ms,(6.10±1.54) ms,and (8.86±0.75) ms,the volatility were (839.81 ± 111.07) μv,(512.25 ±46.82) μv,and (312.65 ±47.52) μv,the NCV were (26.24±3.22) m/s,(21.23 ±2.40) m/s,and (16.34 ±2.55) m/s.Compared with the group C,the peak latency of the SEP,MAP was shortened significantly and the Volatility was increased significantly in group A and B,the NCV was faster significantly in group A and B (P ＜ 0.05).Compared with the group B,the peak latency of the SEP,MAP was shortened significantly and the Volatility was increased significantly in group A,the NCV was faster significantly in group A (P ＜ 0.05).The good rate of the A,B,C groups in BMRC sensory function score were 78.95％,63.15％ and 52.63％ respectively.The good rate in BMRC motor function score were 84.21％,68.42％and 47.37％ respectively.The group A and B were higher than group C,and the good rate of group A was higher than group B(P ＜ 0.05).Conclusion The nerve growth factor gradient release system is safe and effective for early treatment of peripheral nerve injuries.

Traditional Chinese medicine (TCM) is an important part of modern medicine. Application of modern ana-lytical techniques to identify the authenticity of TCM and evaluate its quality is an important and critical content in the process of the modernization and internationalization of TCM. This paper described the development and charac-teristics of a 1H-NMR-based metabolomics technology, and reviewed its application in the species identification, quality evaluation of Daodi Y aocai (i.e., genuine medicinal materials), processing theory and the best harvest time of TCM. Besides, based on previous work, further discussion was given on technical methods and key question of the 1H-NMR metabolomics method. This paper provided a methodological reference for the species identification and quality evaluation of TCM and other herbal medicines.

BACKGROUND: Research has shown that the vascular endothelial growth factor (VEGF) of the ends of the fractured bone is heavily expressed 72 hours to 3 weeks after the fracture and it is supposed that it has a promoting effect on fracture healing. Inducing angiogenesis through VEGF gene transfection has gradually attracted the attention of the researches.OBJECTIVE: To find an efficient way of exogenous VEGF gene in vivo transfection through injecting the mixture of positive ion liposome transfection agent and plasmid and to study the promoting effect of extra VEGF gene expression on bone fracture healing.DESIGN: A randomly grouping, blank control trial.SETTING: Animal Laboratory of Huaxi Medical Center of Sichuan University MATERIALS: Totally 40 adult male SD rats, weighing 230 to 250 g,were involved. All the animals were randomly divided into the experiment group and the control group with 20 rats in each group.METHODS: The experiment was conducted at the Animal Laboratory of Huaxi Medical Center of Sichuan University from April to December 2003.Altogether 40 rats were involved to establish fractured models of right shaft of femur. Cut the bone in the middle of bone stem, retroplanted a Kirsh' nail with 1 mm diameter through intercondylar part and the fractured bone was fixed. In the experimental group, a mixture of 100 μL of liposome transfection agent and 100 μg of pBLAST49-mVEGF plasmid was injected in multiple points into the subperiosteum of the both sides of the ends of the fractured bone. The same volume of normal saline was injected into the rats in the control group. Then, 2 rats in each group were put to death 3,7,14,28,42,56,70 days after the operation and femoral bone specimen was collected.MAIN OUTCOME MEASURES: Observation of right femoral fractured staining results of VEGF, with the apperance of brown granules as positive.RESULTS: Two rats were selected at 7 time points separately, and altogether 28 rats entered the stage of result analysis. The other 12 rats were fracture at different time points: For the experimental group, 28 days after the operation, cartilage callus appeared and replaced fibrocallus gradually,and the fracture line disappeared. Fifty-six days after the operation, the bone healed completely. For the control group, 28 days after the operation , fibrocallus was observed, and the fracture line was still clear. 56days after the operation, much callus appeared, and the fracture line beof fractured bone was stained with hemotoxylin eosin (HE). In the experiment group, 56 days after the operation, the bone healed completely and trabecular like bones were rebuilt. The bone marrow cavity of the fractured region was open again. In the control group, Fifty-six days after the operation, no mature bone was formed, and the bone marrow cavity was not different time points: The expression in the two group reached to the peak on day 14 and began to decrease on day 28. The expression of VEGF in the experimental group was obviously higher than that in the control group.CONCLUSION: Injection of the mixture of positive ion liposome transfection agent into the subperiosteum of rats is an effective approach for in vivo transfection and pBLAST49-mVEGF gene transfection can effectively facilitate the bone fracture healing of rats.

BACKGROUND: Ectogenesis vascular endothelial growth factor (VEGF) could enhance the activity of alkaline phosphatase (AKP) and concentration of cycli adenosine monophosphate (cAMP) fivefolds in cultured osteoblast cell. What' s the effect of ectogenesis VEGF gene transfection on osteoblasts is still by no means clear.OBJECTIVE: To investigate the effect of gene transfection and expression of ectogenesis VEGF on the osteogenesis activities of osteoblast cell.DEDIGN: A completely randomized controlled study.SETTING: Laboratory of Transplantation and Immunity, West China Hospital, Sichuan University MATERIALS: Cranial osteoblasts of newborn two or three-day male SD rat.METHODS: The experiment was conducted in the Laboratory of Transplantation and Immunology of Huaxi Hospital, Sichuan University from April to December 2003 The cranial osteolasts of newborn rat were separated and cultured with enzyme digestion method then were identified by teoblasts cultured in vitro with cation liposomes transfection as gene transations, immunohistochemical staining was performed on VEGF and collagen type I and osteocalcium were detected.collagen I and secretion of osteocalcium of osteoblasts.RESULTS: The concentration of osteocalcium and expression of type I collagen of the 1- 5 generation osteoblast cell in pBLAST49-mVEGF gene transfer group were significantly higher than that in the control group (P ＜0.05).CONCLUSION : It is found in this experiment that the synthesis of collagen I was enhanced obviously after sussceful transfection of pBLAST49-mVEGF plasmid. Compared with the control group, the diffence of intergrated optical density gained by Mias image analysis system was significant( P ＜ 0.05),indicating that pBLAST49-mVEGF plasmid transfection can improve the synthesis of type I collagen and secretion of osteocalcium of osteoblasts.

Objective To investigate the correlations between vascular dementia and neurotrophin-3 (NT-3), fibroblast growth factor (FGF). Methods Female and male Wistar senile rats were divided into the control group and the operation groups at random. The operation groups consisted of the cerebral ischemia 15 min group and ischemia 15 min reperfusion groups in which the rats were reperfused at 0, 1st, 6th hour and on 2nd, 4th, 9th day with five rats per group. The expressions of NT-3 and FGF in hippocampus and thalamus were examined in different periods of ischemia-reperfusion with ABC method of immunohistochemistry. Results There was a little expressions of NT-3 and FGF in hippocampus and thalamus of normal senile rats; the expression of NT-3 in hippocampus increased quickly and steadily from 1 h to 4 d after ischemia-reperfusion! there were significant differences compared with control group (P0. 05). The expression of FGF in hippocampus increased from 6 h to 2 d after ischemia-reperfusion (P 0. 05). FGF in thalamus began to increase on 2nd day after ischemia-reperfusion and there were significant differences compared with control group (P0. 05). Conclusion Hippocampus has the protective mechanism of NT-3 and FGF against ischemia injury which is produced quickly and has endurable effects, but thalamus lacks the good protective mechanism of NT-3 and FGF.

OBJECTIVE: To investigate the feasibility and efficacy of endoscopic sinus surgery for sinonasal malignancy. METHOD: Clinical data of 40 cases of sinonasal malignancy treated in our hospital from January 2007 to December 2011 were retrospectively analyzed. The pathological types encountered were: 5 cases of malignant transformation of sinonasal inverted papilloma, 2 cases of maxillary sinus carcinoma, 3 cases of ethmoid sinus carcinoma, 11 cases of mucosal melanoma, 7 cases of olfactory neuroblastoma, 2 cases of neuroendocrine carcinoma, 4 cases of myoepithelial carcinoma, 3 cases of adenoid cystic carcinoma, 1 case of malignant hemangiopericytoma. These patients were operated on either by an exclusive endoscopic surgery or endoscopic surgery with a complementary external approach. Thirty-two patients received adjuvant radiotherapy and/or chemotherapy. RESULT: All the patients were followed up for a period of 1 to 5 years after operation, of which 18 cases survived for more than three years. Four cases recurred after six months to two years, but the result was satisfactory after revision endoscopic sinus surgery. Three cases occurred cervical lymph node metastasis, of which 2 cases received neck dissection and 1 case received chemotherapy and radiotherapy due to no surgical indications. Four cases occurred distant metastasis. Eight cases died of tumor until the last follow-up. The other 21 cases occurred no recurrence and metastasis during follow-up period. CONCLUSION Endoscopic sinus surgery is an effective method for sinonasal malignancy. Appropriate indication must be carefully selected for malignant tumor. Combined treatment based on surgery should be adopted to it.
Adult ; Aged ; Endoscopy ; methods ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; surgery ; Nose Neoplasms ; surgery ; Paranasal Sinus Neoplasms ; surgery ; Retrospective Studies ; Young Adult