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Since the enrollment in 2002, the cultivation model of "4+4" program of clinical medicine in Shanghai Jiao Tong University School of Medicine has been continuously explored and practiced. With the goal of cultivating high-level compound outstanding medical innovative talents with multi-disciplinary cross-capacity, through strengthening the heuristic teaching, establishing the medical-engineering cross-course, emphasizing the training of scientific research ability, and taking teaching reform in the basic clinical single-circulation organ system integration course, we have established a talent training system with the characteristics of Shanghai Jiao Tong University School of Medicine, which is characterized by "thick foundation, strong practice, re-transformation, shaping norms, and international integration", and intend to make further exploration in the field of post-graduation education convergence.
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Referring to Building Block Innovation concepts, the article addresses the development and current practices of the doctoral education in the intersection of medicine and engineering from modules such as scientific research, tutor guidance, curriculum study, integration of study and industry, and evaluation mechanism. The article proposes inspiration and recommendation to further improve the quality of the doctoral education from dimensions including the management system, the organization system, and the education system.
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Background:MicroRNA-9 (miR-9)is involved in the modulation of a variety of physiological process such as organ development and self-renewal and multipotential differentiation of cells. Moreover,its aberrant expression is closely associated with several types of malignant tumors. Aims:To investigate the effect of miR-9 on biological behavior of human gastric cancer cells. Methods:Expression of miR-9 in normal human gastric epithelial cell line GES-1 and human gastric cancer cell line BGC-823 and SGC-7901 was detected by real-time PCR. Then the two gastric cancer cell lines were transiently transfected with miR-9 mimic,miR-9 inhibitor and empty vector,respectively;the efficiency of transfection was assessed by real-time PCR. CCK-8 assay,Transwell assay and wound healing assay were used to examine the cell proliferation and migration capacities. Results:The expression level of miR-9 was significantly higher in gastric cancer cells than in normal gastric epithelial cells (P＜0. 05). Compared with cells transfected with empty vector,overexpression of miR-9 occurred in cells transfected with miR-9 mimic and effectively promoted the proliferation and migration of BGC-823 and SGC-7901 cells (P ＜ 0. 05 ). Conversely,transfection with miR-9 inhibitor significantly suppressed the proliferation and migration of gastric cancer cells (P＜0. 05). Conclusions:Up-regulating miR-9 expression promotes the proliferation and migration of gastric cancer cells,which indicates that miR-9 overexpression may be associated with progression of gastric cancer.
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Background:There is no specific therapy for inflammatory bowel disease(IBD),and the pathogenesis of IBD is not fully clear. Aims:To explore the expression and clinical significance of IL-17BR in colon mucosa and peripheral blood mononuclear cell(PBMC)of patients with IBD. Methods:Colon mucosal biopsy specimens of 40 Crohn's disease(CD) patients,32 ulcerative colitis(UC)patients and 25 healthy controls and PBMC of 30 CD patients,27 UC patients and 25 healthy controls were collected. The expressions of IL-17BR in colon mucosa and PBMC were determined by immunohistochemistry and flow cytometry,respectively,and correlations between IL-17BR expression and levels of CRP, ESR,CDAI score and Mayo score were analyzed. Serum levels of TNF-α before and after infliximab(IFX)treatment were determined by ELISA,and correlations with IL-17BR were analyzed. Results:Compared with healthy controls,IL-17BR expressions in colon mucosa of CD and UC patients were significantly increased(P<0.05). IL-17BR expressions were significantly higher in active CD and UC patients than in remission CD and UC patients(P <0.05). No significant differences in IL-17BR expression in PBMC were found among CD,UC patients and healthy controls(P>0.05). The expression of IL-17BR in colon mucosa was positively correlated with CRP,ESR,CDAI score or Mayo score in CD,UC patients(P <0.05). After treatment with IFX,expression of IL-17BR in colon mucosa and serum TNF-α level were significantly decreased in CD patients(P<0.01). Expression of IL-17BR was positively correlated with serum TNF-α level in CD patients(P<0.05). Conclusions:The increasing of IL-17BR expression in IBD patients is closely correlated with activity of inflammation and TNF-α level. IL-17BR may play a vital role in immune response of intestinal mucosa,and can be used as a new marker for reflecting the activity of IBD.
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Exosomes are vesicular bodies secreted by living cells containing proteins and RNA, and play an important role in the process of physiology and pathology.Pancreatic cancer is one of the common gastrointestinal tumor with high morbidity and mortality.As a hotspot in oncology, exosomes have potential values in the research of development, diagnosis and treatment of pancreatic cancer.This article reviewed the role of exosomes in pancreatic cancer.
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Biliary tract infection is the most commonly seen severe complication of malignant obstructive jaundice (MOJ)with a high mortality rate. Bile bacterial infection in MOJ patients is related with a variety of clinical factors and the therapeutic approaches including surgical operation,ERCP and percutaneous transhepatic cholangial drainage( PTCD). Bile bacterial culture combined with drug sensitivity test is of great importance for selection of optimal antibacterial agents. In this article,the related factors,current status of therapeutics,biliary bacterial spectrum and drug sensitivity in MOJ with biliary tract infection were reviewed for defining the clinical indicators and guiding the use of antibacterial agents.
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Objective To evaluate the efficacy and safety of photodynamic therapy(PDT)combined with intravitreaIinjection of bevacizumab for choroidal neovascularization(CNV)caused by agerelated macular degeneration(AMD). Methods A total of 21 eyes of 21 patients with AMD,which was diagnosed by examination of visual acuity,intraocular pressure,ocular fundus,fundus color photography,fundus fluoreseein angiography(FFA),indocyanine green angiography(ICGA)and optic coherence tomography(OCT),were underwent PDT combined with intravitreal injection of Bevacizumab.The patients,15 males(15 eyes)and 6 females(6 eyes),aged from 56 to 78 years,with the average of 68.6years.The best corrected visual acuity:counting fingers/10cm-0.9,logMAR was 1.04±0.41.CNV located in below or side central fovea of macula.There was obvious leakage of fluorescein which examined by FFA and ICGA.The average of retinal thickness of macular foveal was(258.91±78.66)μm.The treatment method of PDT has to according to the way of PDT for TAP and Verteporfin PDT for VIP.Intravitreal infeetion with 1.5mg bevacizumab was performed after three days under surface anesthesia.Follow-up time was 1,3,6,12 months after the treatment. Resuits At last visit,the best corrected visual acuity:counting fingers/10 cm-1.5,logMAR was 1.04±0.41,and the differences are statistically significant compared with before.The BCVA improved four or more lines in 6 eyes(28.57%),improved two to four lines in 9 eyes(42.86%),stabilized(±1 line or no change)in 6 eyes(28.57%)and decreased in none.The average intraocular pressure was(15.20±2.41)mmHg after surgery,and the differences was not statistically significant compared with before(P＞0.05).FFA and lCGA showed CNV complete closure in 13 eyes(61.90%).partial closure in 8 eyes(38.10%).The average of retinal thickness of macular foveal was(127.38±20.14)μm(P＜0.01). Conclusion Combining treatment with PDT and intravitreal injection of Bevacizumab is safe and effective for CNV which caused by AMD.It has significant improvement in BCVA.1eakage of CNV and retinal edema.
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BACKGROUND: At present,enhanced green fluorescent protein(EGFP)is proved to be the best labeled molecule,with unique advantages,such as high fluorescence specificity,easy to be detected,and so on.Recombined retroviral vector EGFP-pLNCX2,which can stably express EGFP,can be construct using gene-engineering technique.Transfecting allogenic chondrocytes is indeed very useful to investigate the target gene expression and process of constructing tissue engineered cartilage in vivo.OBJECTIVE:To construct recombined retroviral vector EGFP-pLNCX2 which can stably express EGFP,and investigate optimal conditions for retrovirus transfection of chondrocytes.DESIGN:Randomized controlled observation.SETTING:Shanghai Jiao Tong University.MATERIALS:Thirty New Zealand rabbits of either gender,1 week of age,were purchased from Experimental Animal Center of Chinese Academy of Sciences.Amphotropic retrovirus package cell line PT67 pLNCX2 and pEGEP-C1 were purchased from Clontech Company;NIH 3T3 cell line was purchased from ATCC Company;DH5a Bacterium coli was preserved by the laboratory of Shanghai Jiao Tong University;Retroviral vector pLNCX2 was purchased from Clontech Company;pEGFP-C1 plasmid with EGFP were donated by professor Cong Xiao-qian from Chinese Academy of Sciences.METHODS:This experiment was carried out in the Shanghai Jiao Tong University in August 2005.Chondrocytes of New Zealand rabbits were isolated and cultured.The recombinant retroviral vector EGFP-pLNCX2,which can stably express EGFP,was constructed to transfect cultured chondrocytes from New Zealand rabbits by using gene engineering technique.Transfection results were observed under fluorescence microscope.Altogether 6×105 chondrocytes incubated in 10 cm-diameter flat plate were used to transfect retrovirus-EGFP immediately and at 12,24 and 48 hours after inoculation.One week later,EGFP expression efficiency was measured with flow cytometer,and best occasion for retrovirus transfecting primary chondrocytes.Enzyme-digested chondrocytes were inoculated for 24 hours to transfect retrovirus.After 250 mg/L G418 was added,chondrocytes were screened on the 2nd,3rd,4th,5th and 6th days separately.After phosphate buffer solution(PBS)washings,the transfection efficiency of chondrocytes was detected by flow cytometer,and the best occasion for G418 screening was observed.MAIN OUTCOME MEASURES:①EGFP-pLNCX2 transfection efficiency.②The best occasions for retrovirus transfecting primary chondrocytes and G418 screening.RESULTS: ①Retroviral vector EGFP-pLNCX2 transfected primary chondrocytes of rabbits,and high expression of transfected chondrocytes could be obtained through preliminary screening of G418.After being screened and expressing EGFP,chondrocytes kept normal morphology,with pseudopod adhering to the wall and matrix secreting vigorously.②The best occasion for retrovirus transfecting primary chondrocytes was at 24 hours after cell inoculation.The transfection efficiency determined with flow cytometer was 19.14% on the 5th day.The best occasion for G418 screening was on the 5th day after culture.Transfection efficiency of G418 screening was 55.75% on the 7th day.CONCLUSION:Recombinant retroviral vector EGFP-pLNCX2 can effectively transfect chondrocytes.The best occasion for retrovirus transfecting primary chondrocytes is at 24 hours after inoculation, and the best occasion for G418 screening is on the 5th day after culture.
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BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.
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<p><b>OBJECTIVE</b>To identify the gene mutation of a kindred with type I antithrombin deficiency.</p><p><b>METHODS</b>All of the seven exons and intron-exon boundaries of antithrombin gene were analysed by PCR and direct sequencing of amplified PCR products from the propositus.</p><p><b>RESULTS</b>A 13389G deletion in exon 6 was characterized in propositus, and this mutation led to frameshift.</p><p><b>CONCLUSION</b>This is a novel mutation, which can cause antithrombin deficiency and thrombosis.</p>
Subject(s)
Adolescent , Humans , Male , Antithrombins , Genetics , Base Sequence , DNA , Chemistry , Genetics , DNA Mutational Analysis , Family Health , Frameshift Mutation , Pedigree , Sequence DeletionABSTRACT
<p><b>OBJECTIVE</b>To accomplish a kind of therapeutic gene for hemophilia A, and observe the expression of human factor VIII (hF VIII) in vivo.</p><p><b>METHODS</b>Human clotting factor VIII cDNA with B-domain deleted (Delta760aa approximately 1639aa) was inserted into vector pRC/RSV to form pRC/RSV-hF VIII BD, which conjugated with in vivo liposome transfection reagent (DOTAP-Cholesterol) to accomplish a kind of therapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol. Mice were injected with pRC/RSV-hF VIII BD-DOTAP-Cholesterol i.m. and sacrificed 48 hours, 10 days, 20 days, 30 days, 40 days and 50 days later, respectively. Tissues such as heart, liver, spleen, lung, kidney and muscle were harvested, the distribution and transcription as well as expression of hF VIII BD cDNA were detected by means of PCR, RT-PCR and immunohistochemistry techniques. In addition, the antigen and antibody of hF VIII in plasma were measured.</p><p><b>RESULTS</b>There was high expression of hF VIII in plasma and tissues at the 48(th) hour after injection. On day 10, antigen level of hF VIII in plasma reached its peak, 17.55 ng/ml, and gradually reduced later. The antibody of hF VIII in plasma emerged on day 10 after injection, and increased and gradually reached 37.06 U/ml on day 50 after injection. PCR, RT-PCR and immunohistochemistry showed that hF VIII BD cDNA and its transcription as well as expression existed in all kinds of tissues, and lasted longer in spleen, lungs and kidneys than in heart, liver and muscle.</p><p><b>CONCLUSION</b>Therapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol, produced by combination of pRC/RSV-hF VIII BD and DOTAP-Cholesterol liposome can express human F VIII successfully in vivo, which lays an experimental foundation for curing hemophilia A by gene-drug in clinic.</p>