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We integrate web-based learning with team based learning,which is called WTBL method (Web-based and team-based Learning). WTBL method is constructed and applied to the teaching practice of core curriculum group courses of clinical medicine. We build some small private online courses. The students can preview online and do the case discussions by teamwork in class. The application of WTBL teaching method has realized the flipping of classroom, and helps to enhance students' self-learning ability and teamwork ability.
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Objective:To express and purify the fusion protein of GST-FILIP-1L and prepare its polyclonal antibody. Methods:The constructed recombinant expression vectors pGEX-4T3-FILIP-1L were transformed into Escherichia Coli BL21. FILIP-1L fusion protein was induced by IPTG and purified by Glutathion Sepharse 4B . The rabbit was immunized by the purified fusion protein,and pro-duced serum with anti-FILIP-1L antibody. The titer of polyclonal antibody was detected by ELISA, the anti-FILIP-1L polyclonal antibody was purified by Active and its combining specificity with FILIP-1L protein was further identified by Western blot. Results:The GST-FILIP-1L fusion protein was highly expressed in E. coli, and its specific polyclonal antibody was obtained after the immunization. The polyclonal antibody purified by Active Ester Agrose was able to combine specially with FILIP-1L protein and transformed FILIP-1L protein in 293 cells and FILIP-1L protein of liver cancer cells, respectively. Conclusion: The GST-FILIP-1L fusion protein was expressed successfully,the anti-GST-FILIP-1L polyclonal antibodies with high titer and specificity are successfully prepared,these antibodies provide an useful experimental tool to research the biological function of the FILIP-1L protein.
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Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.
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Objective To Study the relation between analgesic action of alcohol extraction of Natrix tigrina Lateralis and central nervous system. Methods Manifold ache models of experimental nerve centre were established to observe the analgesic action and degree of alcohol extraction of Natrix tigrina Lateralis to the ache models. Results Alcohol extraction of Natrix tigrina Lateralis had analgesic action to all ache models of nerve centre, and the high-dose group was obviously. Conclusions Analgesic action of alcohol extraction of Natrix tigrina Lateralis is related to effecting on central nerves system, which effects antinociceptive in central nervous.
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Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.
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AIM: To investigate the effect of endothelin (ET), angiotensin II (AngII) and homocysteine (Hcy) on C-type natriuretic peptide (CNP) synthesis and release. METHODS: Human endothelial cell was cultured; CNP was measured by radioimmunoassay method. RESULTS: ET and AngII could augment CNP synthesis in human endothelial cells. Compared with control group, 10-9,10-8,10-7 mol/L ET and Ang II increased CNP content of endothelial cells by 1%(P＞0.05), 49%(P＜0.05),117%(P＜0.01) and 137% (P＜0.01),165%(P＜0.01),201%(P＜0.01),respectively. A great dose of ET and Ang II also stimulated CNP release from cultured human endothelial cells. Hcy had no effect on CNP synthesis, but 10-9，10-8,10-7 mol/L Hcy enhanced CNP release from cultured human endothelial cells by 17%(P＞0.05),84%(P＜0.01) and 555%(P＜0.01), respectively. CONCLUSION: ET, AngII and Hcy might be involved in the synthesis and release of human endothelial cell CNP.
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AIM: To observe whether metallothionein plays a role in cardiac protective effect of basic fibroblast growth factor on anoxic/reperfusion (A/R) injury in cultured cardiomyocytes and study the possible mechanism of cardiac protection by bFGF.METHODS: The present study made the model of myocyte A/R injury after having a 24 h incubation by bFGF(10 -10、10 -9、10 -8 mol/L) and bFGF(10 -9 mol/L)+PD 098059 respectively. We measured the levels of MT and MDA in myocytes, and the changes of LDH and protein in cultured medium. We also counted the number of viable cell in groups.RESULTS: The contents of myocardial MT were significantly increased after treatment by bFGF. The levels of MT in 10 -10 mol/L、10 -9 mol/L and 10 -8 mol/L bFGF treated groups increased 54%\, 62%\, 76% respectively, compared with the A/R group, and the number of viable cell were also greatly increased, LDH and protein leakage in cultured medium and MDA contents in myocyte were dramatically decreased in bFGF treated groups. All the protection were completely disappeared with the inhibition of MT production with PD 098059, the inhibitor of mitogen-activated protein kinase(MAPK).CONCLUSION: MT involves in the protection of bFGF in cultured cardiomyocytes. It might be related with activation of MAPKase.
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AIM: To observe the changes of metallothionein (MT) in various tissues of mice during hyperhomocysteinemia. METHODS: Intraperitoneal injection of homocysteine into mice induced hyperhomocysteinemia. The contents of tissue MT and malondialdehyde (MDA) in liver, heart and kidney were determined. RESULTS: Compared with control group, tissue MT levels in Hcy-group animals were increased by 210% ( P 0.05),respectively, compared with Hcy alone group. Tissue MDA contents were decreased by 24% ( P
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AIM: To study the role and regulation of calcineurin(CaN) in angiotensin II(AngⅡ)-stimulated cardiacmyocyte hypertrophy of rats. METHODS: Using AngⅡ to induce the cultured cardiac myocyte hypertrophy of rats, and investigating the effect of CaN inhibitor on [ 3H]-leucine incorporation of AngⅡ-stimulated cardiomyocytes and the regulation of various factors on CaN activity in cardiomyocytes.RESULTS: AngⅡ can stimulate the CaN activity in cultured neonatal rat cardiomyocytes in a dose- and time-dependent manner. In cardiac myocytes incubated with 10, 100, 1000 nmol?L -1 of AngⅡ for 12h, the CaN activities increased respectively by 13%,57%( P
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AIM: To observe the level of metallothionein (MT) in liver, aorta and plasma of rabbit with atherosclerosis (AS) in order to recognize the alteration of oxidative defense system in body when AS occurred.METHODS:Preparation of AS model of rabbit induced by having high-fat diet for eight weeks; the levels of MT and malondialdehyde (MDA) were measured in the tissues of liver and aorta and plasma of rabbit.RESULTS:The MT levels in liver tissues and plasma in atherosclerotic group increased 318%( P
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AIM: To observe whether metallothionein plays a role in cardiac protective effect of basic fibroblast growth factor on anoxic/reperfusion (A/R) injury in cultured cardiomyocytes and study the possible mechanism of cardiac protection by bFGF.METHODS: The present study made the model of myocyte A/R injury after having a 24 h incubation by bFGF( 10-10、10-9、10-s mol/L) and bFGF( 10-9 mol/L) + PD098059 respectively. We measured the levels of MT and MDA in myocytes, and the changes of LDH and protein in cultured medium. We also counted the number of viable cell in groups. RESULTS: The contents of myocardial MT were significantly increased after treatment by bFGF. The levels of MT in I0-l0 mol/L、10-9 mol/L and 10-8 mol/L bFGF treated groups increased 54 %、 62%、 76% respectively, compared with the A/R group, and the number of viable cell were also greatly increased, LDH and protein leakage in cultured medium and MDA contents in myocyte were dramatically decreased in bFGF treated groups. All the protection were completely disappeared with the inhibition of MT production with PD098059, theinhibitor of mitogen- activated protein kinase(MAPK). CONCLUSION: MT involves in the protection of bFGF in cultured cardiomyocytes. It might be related with activation of MAPKase.
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Objective To investigate the effects of a new radiosensitizer, nitroimidazole-derivative--Doranidazole on pancreatic cancer cells under hypoxic condition. Methods The human pancreatic cancer cell lines were exposed in vitro to a single fraction of high dose?-ray radiation either with doranidazole or under hypoxic condition. The percentage of dead cells was analyzed with a multiwell plated reader and the fluorescence intensities of propidium iodide before and after a digitonin tratment were measured. The sensitizing effect of doranidazole on cell killing by high-dose irradiation was evaluated by time-course, dose-dependency, and microscopic observations. The selective radiosensitive effect of doranidazole on hypoxic cells was evaluated by flow cytometry. Results Of seven pancreatic cancer cell lines, the mortalities of 5 cell lines were significantly increased in presence of doranidazole under hypoxic condition 5 days after 30 ?Gy irradiation, but not under aerobic condition. Meanwhile, the radiosensitizing effect of Doranidazole on pancreatic cancer cells was demonstrated as time- and dose-dependent manner. Conclusion Doranidazole can enhance the radiosensitivity on pancreatic cancer cells under hypoxic condition with high-dose irradiation.
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AIM: To observe the role of exogenous and endogenous basic fibroblst growth factor (bFGF) on myocardial ischemia/reperfusion(I/R) injury of rats.METHODS:bFGF and bFGF antiserum were applied to rat isolated I/R heart. Myocardial function, coronary effluent volume,protein and myoglobin content as well as LDH activity in coronary effluent fluid, myocardial calcium, MDA and ATP concentration as well as PKC, MAPK activity were measured. RESULTS:Compared with control, myocardial function in I/R group significantly decreased. Protein, myoglobin content and LDH activity in coronary effluent liquid as well as myocardial MDA and calcium content increased, while myocardial ATP concentration decreased(all P
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AIM: To study the effect of homocysteine (HCY) on proliferation of airway smooth muscle cells and fibroblasts and the effect of HCY on collagen prodution of airway fibroblasts. METHODS: [3H]-TdR incorpora- tion was measured in cultured airway smooth muscle cells. The [3H]-TdR and [3H]-proline incorporation were mea- sured in cultured airway fibroblasts. RESULTS: HCY induced proliferation of airway smooth muscle cells and fibroblasts in a concentration - dependent manner. HCY also induced collagen production of airway fibroblasts in a concentration - dependent manner. The inhibitors of protein kinase C, H7 and polymyxin B, inhibited HCY - induced proliferation of airway smooth muscle cells. CONCLUSIONS: HCY induced proliferation of airway smooth muscle cells and fibroblasts, HCY also induced collagen production of airway fibroblasts. The HCY - induced proliferation of airway smooth muscle cells may be related to the pathway of PKC signal transduction.
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Objective:To determine the sensitivity of SMMC-7721 cells to mDRA-6,the synergistic damage effect of mDRA-6 and adriamycin on human hepatocellular carcimoma cell lines and its possible mechanism.Methods:SMMC-7721 cells were cultured with RPMI1640 medium in regular condition.The morphology was observed by microscope.Cytotoxicity was examined by MTT assay. Apoptosis were detected by flow cytometry.Results:(1)The apoptosis of SMMC-7721 cells could be induced by mDRA-6,1.89 ?g/ml of mDRA-6 cound kill 36% of the cells;(2)Concentration-dependent cytotoxicity of adriamycin was exhibited in SMMC-7721 cells;(3)The combination of mDRA-6 and adriamycin exhibited synergistic effect on SMMC-7721 cells.2 ?g/ml of mDRA6 and 40 ng/ml of adriamycin killed 60%SMMC-7721.Conclusion:MDRA-6 can induce SMMC-7721 cell apoptosis.The combination of mDRA-6 and adriamycin exhibit synergistic effect on SMMC-7721 cells.
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In previous studies, we demonstrated 10 types of lymphocytes in lymph nodes, each exhibited a different fluorescent color by our thioflavine staining method. Among them, 4 types were able to differentiate into plasma cells of the same fluorescent colors. In the present study, different types of lymphocytes were demonstrated in human peripheral blood by their different fluorescent colors after thioflavine staining. The lymphocytes from the venous blood of 50 healthy persons were isolated with Ficoll-Conray solution and E-rosette and EAC-rosette tests and fluorescent staining with thioflavine were performed. Most of the lymphocytes in peripheral blood are small ones with nuclei and cytoplasm showing blue fluorescence and the blue fluorescence of the cytoplasm is paler than that of the nuclei. The nuclei in a part of these lymphocytes have distinct boundaries. The nuclei in another part of these lymphocytes are smaller and with indistinct boundaries and indentation on one side and show dim fluorescence. Other lymphocytes show different fluorescence. Some show blue round nuclei with distinct nuclear membrane, and no color of fluorescence in cytoplasm, but with blue white patches on one side of the nuclei. Some show dark blue nuclei and bright blue cytoplasm and others show orange yellow or orange red nuclei and yellow cytoplasm. In addition, lymphocytes of grayish blue or grayish yellow nuclei and bluish green cytoplasm or lymphocytes of yellowish fluorescence may be seen at times. Very few lymphoeytes of orange red nuclei with nearly no cytoplasm may be seen occasionally.The lymphocytes with blue fluorescence and indentation on one side of nucleus, those with blue nuclei and blue white patches in the cytoplasm as well as those with orange yellow nuclei and yellow cytoplasm can form E-rosettes with sheep erythrocytes. They are T cells. The lymphocytes with distinct boundaries of nucleus, small size and blue fluorescence those with dark blue nuclei and bright blue cytoplasm as well as those with orange red nuclei and yellow cytoplasm can form EAC-rosettes with sheep erythrocytes sensitized by specific antibody and complement. They are B cells. The lymphocytes with blue nuclei and blue white patches may transform into lymphocytes with orange yellow nuclei and yellow cytoplasm under ultra-violet light irradiation. The latter are few in number in the blood but may be progressively increased in number on prolonged observation. They belong to Group Ⅲ of lymphocytes and are mainly located in the paracortical thymus-dependent zone of lymph nodes. The sma ller lymphocytes with blue fluorescence and distinct nuclear boundary may transform into lymphocytes with orange red nuclei and yellow cytoplasm, which are also very few in number in the blood and are also progressively increased in number on prolonged observation. They belong to Group Ⅱ of lymphocytes and constitute the main component of lymph nodules in lymph nodes.