AIM:To investigate the feasibility and specificity of gastric carcinoma gene therapy by utilizing RNA interference (RNAi) to inhibit survivin expression in vitro and in vivo. METHODS:Small interference RNA (siRNA) homologous to survivin was designed. pTZU6+1-siRNA-survivin vector was constructed and transfected into BGC-823 cells. The transplanted BGC-823 tumor in nude mice was established to induce RNAi. The changes of survivin gene expression,tumor cell cycle and cell apoptosis were detected by flow cytometry,RT-PCR,Western blotting,immunochemistry and TUNEL. RESULTS:The expression of survivin was obviously inhibited by RNAi in vitro. The phase of cell cycle indicated the reduction of S phase,while G1/G0 phase increased. Cell apoptosis was obvious. Both the mRNA level and the protein expression of survivin decreased obviously. The tumor size reduced after treated with pTZU6+1-siRNA-survivin vector in vivo. The expression of survivin decreased in siRNA treatment group. In contrast,little change in control group in vitro and in vivo was observed. CONCLUSION:RNA interference down-regulates survivin gene expression,inhibits BGC-823 cell proliferation and induces cell apoptosis with good specificity,which may be a possible new approach for neoplasm gene therapy.

We took such measures in internal medicine practice teaching in five-year program as actualizing special teachers’supervision system,adopting multiple teaching methods (enlightening,case-based,question-based,multimedia),taking interview of teachers and students,and enhancing doctor-patient communication.

In China medical university 8 weeks early exposing to scientific research have been arranged for the students on seven-year program of clinical medicine. The purpose is to let the students understand scientific research procedures based on the tutor's research project. The students gain great achievements. The result of questionnaire shows that the attitudes of the students are positive and 81% of them are satisfied in general. The paper also summarizes the experiences of the practice.

Objective To identify the effects of the subthalamic nuclei(STN)high frequency stimulation(HFS)on striatal and nigral dopaminergic metabolism in rats.Methods The effects of subthalamie nuclei high frequency stimulation(STN-HFS)on striatal dopaminergie metabolism was investigated in free moving rats.Reverse transcriptase polymerase chain reaction(RT-PCR)and Western blot of striatal and nigral tyrosine hydroxyiase(TH)were performed.Results Our data suggest that STN- HFS elevated TH protein(0.99?0.14 vs 0.33?0.08,P

OBJECTIVE: This study was designed to find out the impact of micro-ecological environment on the incidence of allergic rhinitis after developing a model of allergic rhinitis on mice. METHOD: Sixty mice were randomly divided into GF group (n=30) and SPF group (n=30). Mice of GF group were fed in the germ-free environment and mice of SPF group were fed in the specific pathogen-free environment. Then each group were randomly divided into model group (20 mice) and control group (10 mice). Establish allergic rhinitis model in the mice of model group using ovalbumin (OVA) at the age of 6 weeks, observe and score the corresponding symptoms and signs that could been induced. Stain with hematoxylin eosin (HE) staining method for nasal mucosa to observe the morphological changes. Using enzyme linked immunosorbent assay to detect the concentration of IgE, IFN-γ and IL-4 in the peripheral blood serum. RESULT: The chi square test showed that the incidence of allergic rhinithis in the mice of GF group was significantly higher than that in the SPF group (P< 0. 05). HE staining showed that the nasal mucosas of allergic rhinitis positive reaction mice were highly congestive and edematous and had a large number of inflammatory cell infiltration, while there was no abnormal morphology of nasal mucosas in mice with no allergic rhinitis reaction. EOS counting displayed that the number of eosinophilic cells in nasal mucosa of positive allergic rhinitis reaction mice was increased significantly. The concentration of IgE and IL-4 in the serum of positive allergic rhinitis reaction mice was highly increased (P <0. 05), and IFN-γ was significantly decreased (P< 0.05). CONCLUSION The difference of micro-ecological environment may play a key role in the occurrence of allergic rhinitis in mice.
Animals ; Disease Models, Animal ; Environment ; Incidence ; Interleukin-4 ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; Ovalbumin ; Rhinitis ; Rhinitis, Allergic ; etiology

Huang-Qin is a traditional Chinese medicine with antiviral, antioxidant, and anti-inflammatory activities. Its major bioactive compounds are diverse flavone O-glucuronides and glucosides. Although three flavonoid O-glycosyltransferases have been identified from S. baicalensis, this information is not sufficient to elucidate the structural diversity of flavonoid glycosides. In this study, nine glycosyltransferase candidate genes were discovered from S. baicalensis by BLAST analysis and their functions were characterized after heterologous expression. Three new flavone O-glycosyltransferases were able to catalyze the formation of major compounds in S. baicalensis, including baicalin and wogonoside. These enzymes could also utilize exogenous flavones as sugar acceptors. This work further elucidates biosynthetic pathways for Scutellaria flavonoid O-glycosides.

Objective:To determine the immunological effects of a new type adjuvant CTA 1-DD/IgG.Methods:Intranasal im-munization of BALB/c mice with OVA in combination with adjuvant CTA 1-DD or CTA1-DD/IgG.Blood samples were subjected to OVA-specific IgG Abs detection and IgG subclass profiles assessment using ELISA.Splenic plasma cells secreting anti-OVA Abs were detected by ELISPOT.Results:CTA1-DD/IgG showed stronger activity in inducing OVA-specific IgG Abs and splenic Abs-secreting cells compared with CTA1-DD.The enhancement of IgG1,IgG2a and IgG2b Abs were observed,and the ratio IgG2a/IgG1 were >1 in both groups.Conclusion: CTA1-DD/IgG can exert enhanced adjuvant effects compared with CTA 1-DD.Mixed IgG subclasses are induced,indicating a more balanced Th1/Th2 response.

BACKGROUND: At present, the commercial artificial small vascular grafts (diameter < 6 mm) are still unsatisfactory, due to poor blocompatibility and low long-term patency rate. Therefore, finding a vascular substitute with normal biological function and studying construction and function of tissue-engineered blood vessel have become hot topics recently.OBJECTIVE: To construct a novel tissue-engineered blood vessel by rabbit bone marrow mesenchymal stem cells (MSCs) and vascular acellular matrix, and to investigate the biocompatibility and patency rate of tissue-engineered blood vessels.DESIGN, TIME AND SETTING: An in vitro randomized controlled study at level of cytology and histopathology was performed at the Laboratory of Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2006 to June 2008.MATERIALS: The decellularized vascular acellular matrix was obtained by a detergent-enzymatic procedure. MSCs from rabbits were isolated using density gradient centrifugation method and cultured in culture flasks coated with fibronectin. Subsequently, the expanded MSCs were seeded on the decellularized scaffolds, and then co-cultured in the self-made bioreactor to construct the tissue-engineered blood vessels.METHODS: Sixty rabbits were randomly divided into three groups. A1.0-cm abdominal aorta was sheared, and a tissue-engineered blood vessel was transplanted on the abdominal aorta using 8/0 polypropylene thread. Tissue-engineered blood vessel group: Tissue-engineered blood vessel was considered as the transplanted vessel; vascular acellular matrix group:Xenoma artery treated by vascular acellular matrix was considered as the transplanted vessel; xenoma artery group: Fresh xenoma artery was considered as the transplanted vessel.MAIN OUTCOME MEASURES: Immunocytochemical staining was used to identify the cultured MSCs. After 3 months of transplantation, the grafts were retrieved for digital subtraction angiography, pathological test and scanning electron microscope examination.RESULTS: Rabbits MSCs presented a whirlpool-like appearance at 8 days after culture. The immunocytochemistry results were consistent with the phenotype of MSCs. After high proliferation, MSCs were seeded onto the vascular acellular matrix for 12 days,and seed cells attached to well in the lumen of blood vessels. Three months after implantation, the patency rate was 90% of tissue-engineered blood vessel group and 80% of vascular acellular matrix group, which was superior to xenoma artery group (25%). At three months after transplantation, HE staining and scanning electron microscope demonstrated that internal, middle,and external membrane were clearly observed in the tissue-engineered blood vessel group, and the membrane morphology was similar to normal artery. The endothelial cells were covered completely. However, the endothelial cells were not covered completely in the vascular acellular matrix group, while mural thrombosis, mild proliferation of intima, and inflammatory cell infiltration were observed. The intima was thick and necrotic in the xenoma artery group, while lumens were stenotic and accompanied with a certain degree of thrombus organization.CONCLUSION: This study provides a new strategy to develop a tissue-engineered blood vessel with excellent biocompatibility and high patency rate constructed by rabbit MSCs and vascular acellular matrix.

Objective To investigate the expressions of intermedin /adrenomeduliin 2 (IMD/AM2) and its receptor calcitonin receptor-like receptor (CRLR) in the kidney of rats after renal ischemia reperfusion injury(IRI). Methods Male Wistar rats were divided randomly into two groups: sham group and operation group. Renal IRI model was induced by clamping both renal arteries. Blood and kidney were harversted at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h after reperfusion, respectively. Renal histological changes were semi-quantitated. Expressions of IMD and CRLR in the kidney were detected by Western blot, and the content of IMD in serum was measured by radioimmunity at 12 h, 48 h, 72 h after repeffusion. Results Kidneys of renal IRI model rats displayed significant pathologic changes, and the changes were much severer at 48 h after reperfusion. The expressions of IMD and CRLR in kidney were significantly up-regnlated at 12 h, 48 h, 72 h after renal IRI (P<0.01). The level of IMD in serum increased at 12 h, 48 h, 72 h after renal IRI (P<0.05). Conclusion The expressions of IMD and its receptor are up-regulated in the kidney after renal IRI, which may participate in the pathophysiological changes induced by renal IRI.

Objective To discuss the experience in the diagnosis, treatment, complications and follow up of carotid body tumor. Methods All the 25 cases were diagnosised by DSA and CTA. The tumor was resected under carotid adventitial plane in 18 cases, with external carotid artery resection in 4cases, and in 3 cases, internal carotid artery (ICA) and external carotid artery (ECA) were resected simultaneously in which internal carotid artery was reconstructed in two cases including using self vein bypass in one and anastomosis between ICA and ECA in the other. ICA was ligated in the third case. Results No cases died perioperatively. ALL CBTs were treated successfully. Horner syndrome and trachyphonia were relieved after operation. Postoperative trachyphonia, bucking and lingual paralysis developed in 3 cases, and in one case with vagus resection caused dyspnea tracheotomy was performed. The rate of nerves injuries was 12% but no semiplegia and aphasia occurred. Follow up period was from 4 to 90 months (average 44 ±6 months) for 21 cases. The trachyphonia and bucking were improved during follow up but the lingual paralysis persists, and tumor recurred in two cases with one dying. Conclusions CBT treatment should include active surgery, sufficient preoperative preparation and avoiding the postoperative nervous complications.