1.Impact of NK cells on multiple myeloma cell line KM-3
Qian SHEN ; Bojian GE ; Deyan LU ; Wei LU ; Shenghua JIANG
Journal of Leukemia & Lymphoma 2010;19(7):404-406,409
Objective To study the apoptosis of multiple myeloma cell line KM-3 induced by NK cells. Methods WST-1 assay was used to detect the killing effect of KM-3 cells treated with NK cells at different effector(E):target(T) ratio. Flow cytometry was applied to analyze Annexin-V+/PI- apoptotic cells and the mitochondrial transmembrane potential. Results NK cells could significantly kill KM-3 cells in a dosand time-dependent manner (P <0.05). After KM-3 cells- were treated with NK cells for 48 hours, the Annexin-V+/PI- cells were increased obviously in dose-dependence (P <0.05). The Annexin-V+PI- cells were increased in time-dependence when treated with NK cells(E:T ratio at 10:1) (P<0.05). The mitochondrial transmembrane potential of KM-3 cells treated with NK cells were significantly decreased in dose-and time-dependence (P < 0.05). Conclusion NK cell can kill KM-3 cells and induce apoptosis in a dose-and time-dependence manner.
2.Comparison of efficacies of two fixation methods in treatment of complex intra-articular fracture of the distal radius
Naiqiang ZHUO ; Jianhua GE ; Wei SHUI ; Zhengjiang LI ; Xiaobo LU
Chinese Journal of Trauma 2009;25(8):717-720
Objective To compare the efficacies and indications of locking compression plate (LCP) and external fixator plus Kirschner wires in treatment of complex intra-articular fracture of the dis-tal radius. Methods Ninety-eight patients with complex intra-articular fractures of the distal radius were treated with volar LCP or external fixator plus Kirschner wires, the efficacies of which were evaluated by comparing the grasping force and wrist function of the patients. Results All the patients were fol-lowed up for an average of 12.4 months, which showed fracture healing in all the patients. According to the wrist function assessment system of New York Orthopedic Hospital (1990), there was no statistical difference in the efficacy of LCP and external f'lxator plus Kirschner wires in treatment of types C1 or C2 fractures (P > 0.05), while the efficacy of external fixator plus Kirschner wires was significantly superior to that of LCP in treating type C3 fracture (P < 0.05). Conclusions For types C1 or C2 intra-articu-lar fractures of the distal radius, the efficacies of LCP and external fixator plus Kirschner wires are simi-lar, while the efficacy of external fixator plus Kirschner wires is superior to that of LCP in treating type C3 intra-articular fracture of the distal radius.
3.Glycoprotein secreted by Absidia corymbifera is associated with apoptosis of human umbilical vein endothelial cells
Fan GUI ; Youwei WANG ; Ge YANG ; Wei LU ; Jie LU ; Jianwen YANG ; Zhaochun LIN
Chinese Journal of Microbiology and Immunology 2011;31(7):582-587
Objective To partially purify the toxic factor secreted by A. corymbifera and to analyze the mechanism of A. corymbifera-induced human umbilical vein endothelial cell (HUVEC) apoptosis. Methods Glycoprotein secreted by A. corymbifera was purified by Con A Lectin chromatography. The influence of different protein fractions on HUVEC apoptosis was determined by flow eytometer. Both denaturing and nondenaturing deglycosylation of purified glycoprotein was performed and the ability of the protein moiety and carbohydrate moiety to induce HUVEC apoptosis was evaluated respectively. Activation of related caspases during A. corymbifera-induced apoptosis was analyzed by Western blot. The role of caspase-8 and -9 in HUVEC apoptosis was investigated using caspase inhibitors. Caspase inhibitors were used to stop the suppression of HUVEC viability by XTT assay. Results Flow cytometric analysis shows the total protein as well as the glycoprotein fraction of A. corymbifera may induce HUVEC apoptosis in a dose dependent manner. In contrast, similar activity was not observed in the non-glycoprotein fraction. Neither deglycosylated protein nor carbohydrate moiety is able to induce HUVEC apoptosis alone. In the apoptotic signaling pathway, caspase9, caspase-3 and cytochrome C were activated significantly, except caspase-8. Moreover, caspase-9 inhibitor, instead of caspase-8 inhibitor, completely abrogates A. corymbifera-induced HUVEC apoptosis. Caspase9 and caspase-3 inhibitors completely waived the suppression of HUVEC viability by A. corymbifera. Conclusion Glycoprotein secreted by A. corymbifera is associated with HUVEC apoptosis. Intact glycoprotein is essential for the apoptotic progress. Intrinsic apoptotic signaling pathway mediates A. corymbifera-induced HUVEC apoptosis.
4.Absidia corymbifera induces apoptosis of human umbilical vein endothelial cells in vitro
Fan CUI ; Youwei WANG ; Ge YANG ; Wei LU ; Jie LU ; Jianwen YANG ; Zhaochun LIN
Chinese Journal of Microbiology and Immunology 2010;30(11):1025-1030
Objective To analyze the influence of Absidia corymbifera on cell activity of human umbilical vein endothelial cells (HUVEC) as well as the related mechanism. Methods Time course analy sis of the influence of A. corymbifera on cell viability of HUVEC was determined by cell counting after Trypan blue staining. Apoptosis of HUVEC induced by A. corymbifera was observed under fluorescence microscope after treatment with apoptosis detection kit. Time course analysis of HUVEC apoptosis induced by A. corymbifera was detected by flow cytometry quantitatively. Effect of caspase-3 inhibitor on A. corymbifera associated apoptosis was also evaluated at the same time. Activation of caspase-3 inside HUVEC was detected by Western blot. Results A. corymbifera inhibited cell viability of HUVEC in a time-dependent manner by Trypan blue staining. After 12 hours' co-culture, A. corymbifera began to show suppression on cell viability (P =0. 001 ). Fluorescence microscope observation revealed A. corymbifera induced apoptosis of HUVEC instead of necrosis. Flow cytometry analysis showed A. corymbifera induced apoptosis of HUVEC in a time-dependent manner. A. corymbifera began to show obvious effect on apoptosis after 12 h co-culture (P =0.0036). Moreover, A. corymbifera-associated apoptosis was almost abrogated completely by caspase-3 inhibitor. Western blot analysis demonstrated that A. corymbifera triggered the activation of caspase-3 inside HUVEC in a timedependent fashion. Conclusion A. corymbifera induces apoptosis of HUVEC in vitro. Such apoptotic signal is transmitted through caspase cascade reaction.
5.Establishment of two cardiac-specific human cardiac troponin C mutation transgenic mice and comparative analysis
Shan GAO ; Wei CHEN ; Ning LIU ; Wenping GE ; Xiang GAO ; Dan LU ; Lianfeng ZHANG ; Wei DONG
Chinese Journal of Comparative Medicine 2014;(3):67-71
Objective To established cardiac-specific transgenic mice of the cTnC D145E and cTnCG159D and compare the HCM and the DCM.Methods The cTnCD145E and cTnCG159D were generated by site-directed mutagenesis and the transgenic plasmids were constructed by insertion of the mutant genes under the control of α-MHC, which is a myocardium specific promoter.The transgenic mice were generated by microinjection and were all maintained on a C57BL/6J genetic backgroud .The cardiac structure and function of the transgenic mice were compared and analysized by echocardiographic and pathological observation at different ages .Results The cTnCD145E and cTnCG159D transgenic mice were established and developed to HCM and DCM, respectively, with aging.The left ventricular end-systolic volume (ESV) and left ventricular end-diastolic volume ( EDV) decreased and ejection fraction ( EF) and left ventricular end-systolic posterior wall thickness (ESPWT) increased in the cTnCD145E transgenic mice, while EDV and ESV increased and EF and ESPWT decreased in the cTnCG159D transgenic mice at 12 months of age.Conclusions Cardiac-specific human cTnCD145E transgenic mice showed HCM phenotypes , and cardiac-specific human cTnC G159D transgenic mice showed DCM phenotypes , which can be used as different models for comparative study of the pathogenesis of cardiomyopathy .
6.Aortic lumen diameter changes during systolic and diastolic periods: evaluation with ECG-gated computed tomography
Weihang LU ; Xin JIA ; Wei GUO ; Jie LIU ; Yangyang GE ; Wei ZHANG ; Bai HE ; Jianfei DONG
Chinese Journal of General Surgery 2017;32(6):497-500
Objective To characterize the changes in the dimensions during systolic and diastolic periods in the aorta with ECG-gated multi-detector CTA scans.Methods The CT angiograms of 115 patients (78 males,mean age 55.2 ± 9.4 years;37 females,mean age 60.1 ± 8.5 years) both in systolic and diastolic periods were obtained on a 64-slice ECG-gated multi-detector CT scanner.The diameters were measured at four anatomic levels of the aorta.(Level A:1 mm proximal to the innominate artery;Level B:1 mm distal to the left common carotid artery;Level C:1 mm distal to the left subclavian artery;Level D:10cm distal to the left subclavian artery).On each level,the maximal and the minimal diameters were measured both in systolic and diastolic periods.Results The paired sample t test results showed a significant difference between the systolic and diastolic diameters in all individual subjects on every level (P <0.001).The mean maximum diameter changes were 1.95% (range-2.0% to 7.0%),2.12% (range-3.0% to 6.0%),1.88%(range-1.0% to 8.0%)and2.47%(range-3.0% to 10.0%)at level A,B,C and D,respectively.The mean minimum diameter changes were 1.43% (range-3.0% to 5.0%),2.67% (range-2.0% to 11.0%),1.75% (range-14.0% to 9.0%)and 2.99% (range -2.0% to 11.0%) at level A,B,C and D,respectively.Conclusions The differences of the aortic diameters between systolic and diastolic periods are significant.The pulsatility of aorta in Chinese population may be different from published Western literature.
7.Effect on the Synthesis of 1,3 propanediol of Organic acids in the Two-stage Fermentation
Ji-Hong LU ; Ge-Bin ZHU ; Wei SHEN ; Zhi-Ming RAO ; Hui-Ying FANG ; Ge-Jian ZHU ;
China Biotechnology 2006;0(05):-
Effect of organic acids on the synthesis of 1,3 propanediol was studied.The adsorption of organic acids from glycerol fermentation liquor by ion-exchange resins was investigated.The results showed that organic acid and 1,3 propanediol production was in negative relationship.The static adsorption showed that ion-exchange resin 005 had the best adsorption abilities of the organic acids in the glycerol fermentation liquor.It was showed that the yield of 1,3propanediol increased by 166% after the extraction of organic acids from glycerol fermentation liquor and the convertion rate increased by 34%.
8.The effects of arsenic trioxide eluting stents on expression of inflammatory factors in injured coronary arteries in pigs
Wenjie TIAN ; Wei YANG ; Junbo GE ; Hao LU ; Li SHEN ; Feirong GONG ; Keqiang WANG ; Yunzeng ZOU
Chinese Journal of Geriatrics 2009;28(2):152-155
Objective To observe the effects of eluting stents coated with arsenic trioxide(As2O3)and suspended in poly-L-lactic acid(PLLA)on expression of monocyte chemoattractant protein-1 (MCP-1)and interleukin-6(IL-6)and to assess the effects of As2O3 eluting stents on local inflammatory reaction in injured coronary arteries in pigs. Methods Bare metal stents,rapamycin eluting stents and As2O3-eluting stents were randomly and double-blindly implanted into the anterior descending branches,circumflex branches and right coronary arteries in eight pigs.Animals were sacrificed and coronary arteries were isolated 7 days after stents implantation.The expression levels of protein and mRNA of MCP-1 and IL-6 were determined by Western blot analysis and reverse transcription polymerase chain reaction(RT-PCR),and the inflammatory cell infiltration was observed by HE staining and immunohistochemistry. Results Compared to bare metal stents,As2O3-eluting stents and rapamycin-eluting stents identically and markedly inhibited the protein expression level of MCP-1(0.421±0.055 and 0.406±0.042 vs.0.857±0.053,P<0.01)and IL-6(0.151±0.032 and 0.146±0.051 vs.0.551±0.032,P<0.01)and correspondingly lowered the mRNA expression level of MCP-1(0.338±0.047 and 0.327±0.051 vs.0.724±0.027,P<0.01)and IL-6(0.531±0.052 and 0.523±0.061 vs.1.015±0.041,P<0.01),and significantly reduced the inflammatory cell infiltration of injured coronary arteries in pigs. Conclusions As2O3-eluting stents can effectively inhibit the expressions of MCp-1 and IL-6 and reduce the inflammatory cell infiltration of injured coronary arteries in pigs.
9.Biochemical markers of myocardial injury in patients of prolonged seizures after modified electroconvulsive therapy
Huabin FU ; Xiaodong ZHOU ; Zhiyuan LIU ; Wei LU ; Ge QI ; Feng CHEN ; Keyong LUO ; Weidong NING
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(2):134-136
Objectiye To observe the prevalence of prolonged seizures and the changes of biochemical markers of myocardial injury in patients with prolonged seizures after modified electroconvulsive therapy(MECT).Methods Patients treated with MECT or simulated ECT were divided into three groups.Group Ⅰ , 26 patients,experienced at least one prolonged seizure after MECT;group Ⅱ,41 selected patients, had not prolonged seizures at all during a course of MECT treatments and group Ⅲ, 31 patients, received simulated ECT.Biochemical markers of myocardial injury, including phosphocreatine kinase (CK), MR isoenzyme of phosphocreatine kinase (CK-MB), lactate dehydrogenase ( LDH ), α-hydroxybutyrate dehydrogenase ( α-HBDH ) and cardiac troponin (cTnT) ,were measured immediately, 3 hours later and on the following day after the first prolonged seizure for group Ⅰ ,the same time points as group Ⅰ after the first treatment of MECT for group Ⅱ , immediately after simulated ECT for group Ⅲ.These indexes were compared between the patients of three groups.Results The positive rate ofcTnT was 30.8%(8/26) and 17.1% (7/41)in group Ⅰ and Ⅱ respectively, but no difference was found(P>0.05 ).CK measured immediately after MECT in patients of group Ⅰ was significantly higher than that of group Ⅲ(P < 0.05 ).CK-MB (immediately), LDH ( immediately and 3 hours later) and α-HBDH ( immediately, 3 hours later and on the following day) in patients of group Ⅰ were significantly higher than those of group Ⅱ and Ⅲ measured after MECT or simulated ECT(P<0.05 ).Conclusion More attention should be paid that absolute or relative hypoxemia may lead to minor myocardial injury.
10.Site-directed mutagenesis of human IL-29 and antineoplastic activity of the recombinant human IL-29 variant.
Wei CHEN ; Rong ZHU ; Chunlei GE ; Yuan LU ; Liyun LI ; Fei LI ; Minchen WU
Chinese Journal of Biotechnology 2015;31(5):702-710
To explore the anti-tumor proliferation activity of human interleukin-29 (hIL-29) variant and based on bioinformatics analyzed data of hIL-29, a mutant gene hIL-29(mut33,35) was amplified by site-directed mutagenesis and megaprimer PCR. The hIL-29(mut33,35) was inserted into an eukaryotic expression plasmid pPIC9K and successfully expressed in Pichia pastoris GS115. A recombinant variant protein (rhIL-29(mut33,35)) was purified from the ferment supernatant of the engineering GS115. To observe the antineoplastic activity of the variant rhIL-29(mut33,35), a CCK-8 reagent was used to detect the anti-proliferation effect. Results show that it has strong anti-proliferation effect when acted on liver cancer cell BEL7402, colon cancer cell HCT8 and gastric cancer cell SGC7901. The inhibition ratios of the three tumor cells were (30.99 ± 1.58)%, (22.47 ± 1.37)% and (32.05 ± 2.02)%, respectively. In high dose group, the anti-proliferation effect of the rhIL-29(mut33,35) was stronger than that of wild type rhIL-29 (P < 0.01). This indicates the variant rhIL-29(mut33,35) has potential development value for medicine.
Antineoplastic Agents
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pharmacology
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Carcinoma, Hepatocellular
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pathology
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Cell Line, Tumor
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drug effects
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Humans
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Interleukins
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biosynthesis
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pharmacology
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Liver Neoplasms
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pathology
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Mutagenesis, Site-Directed
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Pichia
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Plasmids
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Polymerase Chain Reaction
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Recombinant Proteins
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biosynthesis
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pharmacology