AIM: Recombinant adenovirus possesses high transfection efficiency and wide host range. This study was designed to construct the recombinant adenovirus vector containing human vascular endothelial growth factor 165 (VEGF165), so as to lay a foundation for the subsequent gene transfection, microencapsulated genetically engineered cells and animal experiments. METHODS: The experiment was conducted in the Laboratory of Cardiothoracic Surgery (the National Key Laboratory), Changhai Hospital of The Second Military Medical University of Chinese PLA from January to May in 2007. Experiment materials: pAxCAwt.VEGF165 was provided by Institute of Cardiothoracic Surgery of Changhai Hospital. pAxCAwt.VEGF165 and DNA-TPC were cotransfected into human embryonic kidney 293 cells by lipofection method. Being propagated, recombinant replication-deficient adenovirus named Ad.VEGF165 was obtained. The target gene of recombinant adenovirus was identified by polymerase chain reaction (PCR) and restriction enzyme digestion. The titer of virus was detected by 50% tissue culture infective dose method. RESULTS: Construction of recombinant adenovirus Ad.VEGF165: The pAxCAwt.VEGF165 and DNA-TPC were successfully cotransfected into human embryonic kidney 293 cells by lipofection method, and replication-deficient adenovirus vectors coding for VEGF165 gene were generated. Identification of recombinant adenovirus Ad.VEGF165: Two fragments of PCR products (597 bp and 146 bp) were obtained by NcoI restriction enzyme. The result was consistent with that calculated with Gene Tool software. The virus titers was 2.2?1015 pfu/L. CONCLUSION: DNA-TPC and pAxCAwt.VEGF165 can be used to construct replication-deficient recombinant adenovirus Ad.VEGF165 in a high titer, low toxicity, high efficiency and safe transfection in vitro.

[Objective]To explore the reason of ulnar impaction syndrome misdiagnosed by tissue injury in wrist.[Method]From December 1998 to December 2005,216 cases who were diagnosed for soft tissue injury in ulnar wrist were checked and discriminated according to X-ray and MRI of the injured wrist,and 48 cases were re-diagnosed for ulnar impaction syndrome.The misdiagnosis rate was 22.2%.[Result]Most cases of ulnar impaction syndrome had positive lunar variance(68.8%).Carpal avascular necrosis were found in about 27.1% of cases through X-ray of wrist and 100% through MRI.Different degree changes of abnormal signal intensity were found in soft tissue,triangular fibro cartilage(TFC) and carpal bones by MRI which displayed swelling of soft tissue in carpal joint,strip-shape high signal intensity or mixed signal intensity at ligament around wrist and enhanced signal intensity at TFC area extending to ulnar carpal articular facet.The appearance of MRI in carpal bones showed local abnormal signal,articular cartilage thickening and hydropsia in bone below cartilage and marrow.[Conclusion]The misdiagnosis can be reduced through elevating acquaintance level of ulnar impaction syndrome basing on clinical symptoms,making the most use of imaging,especially MRI.

Objective To asses the clinical applied value for the salpingian recanalization by means of interventional procedure.Methods 80 cases ranged in age from 23～35 with sterility diagnosed by uterosalpingography were included in this study.They had bilateral and unilateral salpingian narrow,adhesion or stenosis.According to the interventional radiologic theory,using same-axis catheter system,and recanalization fluid the narrow,stenosis and adhesion of salpingian were treated.Results There were 113 salpingians suffered with narrow,adhesion and stenosis in 80 cases.Of them,87 salpingians were recanalized.The successful rate was 84.5%,among them the recanalization rate was 94.8% in the patients with interstitial lesion and 25 cases had been pregnancy (approximate 31.9%) after operation. Conclusion The recanalization of salpingian by interventional radiology in the treatment of sterility caused by salpingian stenosis,narrow and adhesion is an effective means.The pregnancy rate is relevant to the selection of indication.

Objective To investigate the expression of calpains and calpastatin in diaphragm of a rat model of chronic obstructive pulmonary disease(COPD). Methods Fotry male Wistar rats were randomly divided into the control group and the COPD group. Rats in the COPD group were copied by expo -sing to cigarettes smoking and dripping lipopolysaccharide into trachea. Pathological changes of lung and diaphragm were deteted. Immunohis-tochemistry and RT-PCR were performed to determine protein and mRNA expression of calapins and calpastatin. Results Diaphragmatic atrophy was observed in rats in the COPD group instead of the control group. The protein and mRNA expression of calpains in diaphrag-m of COPD group were Increased than those in the control group(P < 0.05). On the contrary ,the expression of calpastatin was significantly reduced in rats in the COPD group(t =-9.38 and-2.97;P < 0.001 ,P = 0.005). Conclusions An imbalance in the expression of calpains/calpastatin in diaphragm was observed in COPD rats ,and this imbalance may be responsible for diaphragm atrophy.

To study the effect of Buyang Huanwu Decoction (BHD) on neural cell apoptosis after cerebral ischemia in gerbils and its mechanism, gerbil model with cerebral ischemia was established by clamping of bilateral carotid arteries and then reperfusion. Neural cell apoptosis of hippocampal gyrus slides was observed under electron microscope 120 hours after reperfusion. The activities of Na + -K + ATPase , Ca 2+ ATPase , Mg 2+ ATPase and NOS, and the contents of lactic acid, NO and amino acid in cerebral tissues were detected 120 hours after reperfusion. The results showed that BHD can reduce the incidence of neural cell apoptosis of hippocampal gyrus, prevent the decrease of Na + -K + ATPase and Ca 2+ ATPase activities, increase NO content and NOS activity, and reduce cerebral glucose content. It is indicated that BHD can counteract neural cell apoptosis after cerebral ischemia in gerbils and its mechanism may be related to the improvement of cerebral energy metabolism, regulation of NO synthesis and antagonism of toxic effect of excitatory amino acid.

Famous TCM gynecological experts in the contemporary era represent the highest level of TCM gynecology during modern age.This article studies the academic thoughts of eight famous TCM gynecologic experts according to academic schools.The purpose is to seek the commonness of these academic thoughts and research their characteristics.It is extremely significant in inheriting and developing these doctors′ academic thoughts without sectarianism,which will help to guide the clinical works.

Objective To establish mice model that induces mucosal immunity by orally infected tachyzoites of Toxoplasma gondii. Methods Respectively, 5?103, 5?104, 5?105, 5?106 tachyzoites of RH strain were inoculated to BALB/c mice by stomach delivery. The control group was given PBS solution. Symptoms and pathological changes of mice were observed. Secretory im-munoglobulin A (SIgA) was assayed. The lymphocytes from Peyer's patches (PP) and intraepithe-lial lymphocyte (IEL) were observed and the changes of CD4+ and CD8+ T cells assayed by SABC immunohistochemistry. Results Inoculation of 5?104 tachyzoites of Toxoplasma gondii caused symptoms and pathological changes in mice. The titre of SIgA increased in intestine, and CD4+ T subset of the mucosal inductive sites and CD8+ T subset of the mucosal effectors' sites increased. Conclusion Mucosal immunity may be induced by oral infection of 5?104 tachyzoites of T. gondii RH strain in BALB/c mice.

[Summary] Type 2 diabetic rat model accompanied by insulin resistance was established by high fat/high glucose diet and streptozotocin.Following metformin treatment for 4 weeks,realtime PCR and Western blot were used to detect the expressions of SIRT3 mRNA and protein in skeletal muscle tissue,respectively.The results showed that insulin sensitivity,superoxide dismutase (SOD),and glutathione (GSH) levels were significantly reduced in diabetic group compared with the control group (P＜0.05),while methane dicarboxylic aldehyde (MDA) level was increased (P＜0.05),along with the decreased expressions of SIRT3 mRNA and protein in skeletal muscles (P＜0.01).After metformin treatment,the insulin sensitivity,SOD,GSH,and SIRT3 mRNA and protein levels were significantly increased (P＜ 0.05 or P＜ 0.01),while MDA level was decreased (P ＜ 0.05),suggesting that metformin may ameliorate insulin resistance via upregulating SIRT3 expression in skeletal muscles of diabetic rats.

Objective: To investigate the reversal effects of pulsed magnetic fields(PMF) on the drug resistant cell line K562/ADR in vitro and their mechanism.Methods: MTT assay was used to determine the IC50 of ADR on K562/S and K562/ ADR cell lines,the detriment of PMF on the K562/ADR cell line and the reversal effects of the drug resistance after treatment with different frequencies of PMF.Flow cytometry(FCM) was applied to the assessment of concentration of intracellular ADR.Results: The drug-resistance multiple of the K562/ADR cell line was 29.36.PMF did not affect the growth of the K562/ADR cell line.Different frequencies of PMF produced different effects on the IC50 of the K562/ADR cell line.In the same condition,70Hz PMF produced the most significant effect and increasea the concentration of intracellular ADR.Conclusion: PMF can reverse the multidrug resistance of K562/ADR by enhancing the drug concentration in K562/ADR cells,and its frequency is correlated with its reversal effect:low fregucncy produces better effect.

Background and purpose:The morbidity of the elderly patients with hepatic portal cholangiocarcinoma is rising. Due to the characteristics of pathology,physiology and anatomy of the disease, most of the patients with cancer are not resectable, the main treatment consists of the diverting drainage and postoperative chemoradiation.We studied the clinical features and the effective therapeutic method for the elderly patients with hepatic portal cholangiocarcinoma. Methods:A retrospective analysis was performed in 28 cases of the elderly patients with hepatic portal cholangiocarcinoma who were treated between January 1995 and December 2005 in our hospital.Results:3 cases received surgery, 2 of them survived for over three years, another 3 cases were given operative exploration with 14 months of medium survival time. 22 cases were given internal or external drainage and received radiotherapy after operation, their survival time ranged from 5 months to 40 months.Conclusions:Surgery is the primary therapeutic method for elderly patients with hepatic portal cholangiocarcinoma. Internal or external drainage and radiotherapy could prolong survival time of the patients with unresectabke disease.