1.Mannnitol colonic lavage in one-stage resection and primary anastomosis of left hemicolon obstruction
Xingtang GONG ; Haifeng LIU ; Dejun ZHOU
International Journal of Surgery 2009;36(9):643-644
Objective Summarized the therapeutic effect of colonic irrigation by mannitol in one-stage resection and primary anastomosis for left hemicolon obstruction. Method For 31 cases of left hemicolon obstruction in our hospital, After process was cut, firstly we had done coloclysis with normal saline, metron-idazole and 20% mannitol,then one-stage anastomosis. Results With the exception of three cases of in-cision infection, the rest who start eating 4~5 days after operation were cured and no anastomotic fistula. Conclusion During the colon resection in left hemicolon obstruction, coloclysis with normal saline, met-ronidazole and 20% mannitol, then one-stage anastomosis, the intestinal function were quick recovery and can prevent colonic anastomotic fistula.
2.Transfecting hyperpolarization-activated cyclic nucleotide-gated channel 2 gene into porcine bone marrow mesenchymal stem cells
Yinglong YAO ; Hao ZHANG ; Dejun GONG ; Zhigang SONG ; Zhiyun XU
Chinese Journal of Tissue Engineering Research 2009;13(49):9673-9676
BACKGROUND: Hyperpolarization-activated cyclic nucleotide-gated (HCN) current plays an important role in regulating heart spontaneous pulsation.OBJECTIVE: To observe target gene expression and electrophysiological characteristics of pig bone marrow mesenchymal stem cells (BMSCs) transfected with hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) gene recombinant adenovirus.DESIGN, TIME AND SETTING: Cell-gene in vitro study was performed at the Laboratory of Thoracic and Cardiovascular Surgery,Second Military Medical University of Chinese PLA from July 2007 to March 2008.MATERIALS: Yorkshire pig was supplied by Animal Institute, Second Military Medical University of Chinese PLA. HCN2 plasmid was presented by Professor Dario DiFrancesco from Italy. Recombinant adenovirus Ad.HCN2 was constructed and stored using Ad5 in this laboratory.METHODS: Pig BMSCs were isolated with combination of gradient centrifugation of Percoll and adherent treatment in vitro.Ad.HCN2 was transfected at multiplicity of infection=50. We also set non-transfection and transfected Ad.Null groups.MAIN OUTCOME MEASURES: Expression of HCN2 mRNA was detected with RT-PCR, and expression of HCN2 channel protein was examined with immunofluorescent staining. Electrophysiology of HCN2 channel protein was measured with whole-cell patch clamp.RESULTS: No amplified fragments were found in the non-transfection and transfected Ad.Null groups, but amplified fragments were determined at 250-500 bp following Ad.HCN2 amplification, which was the same as plasmid carrying HCN2 gene. Staining strength of cell nuclei following transfection was significantly weaken compared with cell membrane and plasma, which showed identical distribution as HCN2 protein. No HCN2 protein was detected in the non-transfection and transfected Ad.Null groups.Pacemaker current could be recorded with a whole-cell patch clamp. It was fully activated around -140 mV with an activation threshold of -60 mV, presenting voltage dependence. CsCI (4 mmol/L) reversibly blocked the inward currents. No pacemaker current was detected in the non-transfection and transfected Ad.Null groups.CONCLUSION: The HCN2 recombinant adenovirus carrier was transferred into serial subcultivation porcine bone marrow mesenchymal stem cells. HCN2 channel protein has been expressing. Pacemaker current could be recorded with a whole-cell patch clamp.
3.Transplantation of autologous bone marrow mesenchymal stem cells transfected by angiogenin gene in a porcine chromic ischemic heart model
Sengdong HUANG ; Xiaohong LIU ; Yang YUAN ; Dejun GONG ; Zhiyun XU
Chinese Journal of Thoracic and Cardiovascular Surgery 2008;24(6):396-398
Objective To investigate the effect of the transplantation of autologous marrow mesenchymal stem cells transfected by angiogenin gene in a porcine chronic ischemic beart model.Methods Methods Mesenchymal stem cells were trnsfected byAd/Ang.The pigs underwent placement of amerold occluder around LCx and 4 weeks later sujected to transplantation of the trandected mesenchymal stem cells.The animals were evaluated by coronary angiography,echocardiography,mannetic resonance imaging end pathologic observation.Results All animal showed 95%occlusion fo LCx.4 weeks after treatment ,the perfusion of LCx,left ventricular ejection fracton were greatly evhanced.A large number of labeled mesenchymal stem cells were successfully uncorporated into blood vessels in the ischemic myocardial regions with increased vessels countin and survival of implanted cell.Conclusion Transplantaton of autologous mesenchymal stem cells trendected by angiogenin gene offere obvious advsntases of great improvement of blood supply and the heart funcition.
4.The CT Manifestations of Giant Cell Tumor of Bone
Xiangyang GONG ; Guangzhao YANG ; Dejun ZHANG ; Senhua LI
Journal of Practical Radiology 2000;0(12):-
Objective To analyse CT signs of giant cell tumor of bone.Methods CT appearances of 42 cases of giant cell tumor of bone were reviewed.All patients were pathological proven.Results (1)Two kinds of bone destruction were seen in all ases:expansive cystic destruction and osteolytic destruction.The anterior was predominantly.(2)Most of our patients haven't bone septations in their tumors (95.2%).(3)Bone crests were usually seen in our study(63.2%).(4)Complete or incomplete sclerotic borders existed in 44.7% of all cases.(5)The inner bone soft tissues were heterogeneously in most of the patients.Fluid-fluid level can be seen in 5 cases (11.9%).(6)The outer-bone soft tissue masses were demonstrated in the majority cases with discontinuous bone shell.Conclusion CT may deepen our understanding and add imaging signs about giant cell tumor of bone;Which can enhance our diagnostic and differential diagnostic capability.
5.MiR-203 inhibits proliferation and invasion of esophageal squamous cell carcinoma
Yang YUAN ; Dejun GONG ; Xiaohong LIU ; Fanglin LU
Academic Journal of Second Military Medical University 2000;0(08):-
Objective To investigate the influence of miR-203 on the proliferation and invasion of human esophageal squamous cell carcinoma cell line Eca109.Methods Double-stranded mimics of miR-203 were designed and transfected into Eca109 cells with Lipofectamine 2000;Eca109 cells transfected with nonsense microRNA mimics were taken as control.The proliferation ability of Eca109 cells was determined by calculating the cell population doubling time and the percentage of apoptotic cells;the invasion ability of Eca109 cells was determined by Transwell assay.Results In vitro experiment showed that,compared with the control group,Eca109 cells transfected with miR-203 mimics had a significantly longer cell population doubling time([26.1?0.5]h vs [24.2?0.6]h,P
6.Modification of bioreactor for tissue-engineered heart valve and its application
Guangjie HOU ; Zhiyun XU ; Shengdong HUANG ; Dejun GONG
Academic Journal of Second Military Medical University 1982;0(01):-
Objective:To modify the pulsatile bioreactor we constructed previously for simulating the high-flow, high-pressure hemodynamics of heart valve in vivo, and to experimentally cultivate the tissue-engineered heart valves (TEHV) in the modified bioreactor. Methods: T-PLS system (NewheartBio Co., Ltd Korea) was used to generate pulsatile flow in the modified bioreactor and we designed a new air-exchange pathway to avoid contamination. The TEHV were made by seeding human bone mesenchymal stem cells (BMSCs) on de-cellularized porcine heart valve. After cultured under static condition for 4 d, the TEHVs were moved to the modified bioreactor and exposed to low-flow (0-600 ml/min) or high-flow(0-4 800 ml/min) pulsatile hydrodynamics for 7d, then the cells on TEHVs were evaluated. Results: After modification, the flow range expanded from (0-1 200) ml/min to (0-6 000) ml/min and the pressure range expanded from (0-40) mmHg to (0-180) mmHg. In culture experiments, 26.3% of the seeded cells remained under low-flow environment and cells were completely lost under the high-flow dynamics. Conclusion: The modified bioreactor can basically simulate the dynamics of heart valve in vivo and can be used in TEHV cultivation research. However, the current TEHV can not tolerate the high-flow pulsatile hydrodynamics.
7.The role of platelet in mouse pulmonary microvascular endothelial cell injury induced by lipopolysaccharide-activated neutrophil in vitro
Zhenzhen SUN ; Jiafeng WANG ; Dejun GONG ; Jinjun BIAN ; Keming ZHU ; Xiaoming DENG
Chinese Journal of Anesthesiology 2011;31(11):1359-1362
ObjectiveTo investigate the role of platelet in mouse pulmonary microvascular endothelial cell (PMVEC) injury caused by lipopolysaccharide( LPS)-activated neutrophil.MethodsPMVECs were obtained from pathogen-free C3H/HeN mice of both sexes aged 6-8 weeks weighing 18-25 g according to the method described by Lim YC et al.Platelets and neutrophils were isolated from mouse blood by twice centrifugation and denaity gradient centrifugation respectively.PMVECs were seeded into twelve- or six-well plates ( 1 or 2 ml/well) after 2-5 passages and were randomly divided into 4 groups (n =31 each): group LPS; group platelets (group P);group neutrophils (group N) and group platelets + neutrophils (group PN).Each well contained about 5 × 107/ml platelets and/or 5 × 105/ml neutrophils respectively.PMVECs were incubated with LPS1 μg/ml at 37 ℃ in a 5% CO2 humidified atmosphere for 1,6,12,18 and 24 h respectively in all 4 groups.The cells were examined with phase contrast microscope for morphological changes and survival condition.Viability rate,apoptotic rate and activation rate of PMVECs were detected by flow cytometry at each time points.ResultsThere was no significant difference in morphology and number of endothelial cells (ECs) among the 4 groups,while the number of activated ECs was significantly increased but the number of living cells decreased in group PN compared with group LPS.The activation rate of ECs was significantly higher after being incubated with LPS for 6-12 h in groups P and N than in group LPS.The viability rate was significantly lower,while the apoptotic rate and activation rate were significantly higher after ECs were incubated with LPS in group PN than in groups LPS,P and N.ConclusionPlatelets play a decisive role in mouse PMVEC injury induced by LPS activated neutrophils.
8.Immunogenicity of decellularized porcine heart valve
Qiuze LI ; Zhiyun XU ; Shengdong HUANG ; Baoren ZHANG ; Lixin YANG ; Xiaohong LIU ; Yang YUAN ; Dejun GONG
Chinese Journal of Tissue Engineering Research 2009;13(38):7498-7502
BACKGROUND: It has been found that xenogenic extracellular matrix (ECM) may cause a strong inflammatory response in humans during clinical application of decellularized porcine heart valve (synergraft valves). An early inflammatory reaction severely weakens matrix structure of valve wall, leading to structural rupture and decay of grafts. From Synergraft's event, the decellularized porcine heart valves still had immunogenicity, especially for pediatric patients. The mechanisms by which the ECM triggers this immune process need to be further evaluated. OBJECTIVE: To find the difference of gene sequence between human and porcine ECM and to identify the ECM immunogenicity based on bioinformatics. DESIGN, TIME AND SETTING: A contrast study between human and porcine ECM based on type IV collagen was performed at the Laboratory of Cardiothoracic Surgery, Changhai Hospital, the Second Military Medical University of Chinese PLA from June 2008 to February May.MATERIALS: The fresh porcine heart valves were obtained from Shanghai Wufengshangshi Slaughter House. Decellularized porcine aortic valves, hybridoma cells, and monoclonal antibodies were provided by our laboratory. METHODS: Similar region and conservative site of gene sequence among human, porcine, and rat were compared so as to look for common similar region, site, and sequence difference and investigate the segment which caused common and different gene sequence. Type IV collagen monoclonal antibody was used to evaluate the persistence of ECM of decellularized porcine heart valve following immunohistochemical staining. MAIN OUTCOME MEASURES: Type IV collagen gene sequence; efficacy of self-made antibody using immunohistochemistry; effect of self-made antibody on type IV collage of decellularized porcine heart valve. RESULTS: The differential gene serial in type IV collagen protein was found out by bioinformatics method. Monoclonal antibodies were successfully produced by human-mouse hybridoma technique. Residual porcine ECM was observed on decellularized porcine heart valve. CONCLUSION: Residual porcine ECM was observed on decellularized porcine heart valve and had immunogenicity.
9.Exercise Preconditioning Improving the Pathological Cardiac Hypertrophy in Pressure Over-loaded Rats
Tongyi XU ; Qingqi HAN ; Ben ZHANG ; Dejun GONG ; Yang YUAN ; Chengliang CAI ; Yun DING ; Liangjian ZOU
Chinese Circulation Journal 2014;(9):728-732
Objective: To explore the effect of exercise preconditioning (EP) on pathological cardiac hypertrophy and heart failure (HF) in pressure over-loaded experimental rats.
Methods:A total of 60 SD rats at the age of 6 weeks were randomly divided into 3 groups, n=20 in each group. Sham-operation group, Transverse aortic constriction (TAC) group and EP + TAC group. The cardiac function and structure were evaluated by echocardiography, patholgical changes and HF biomarkers were examined for EP effect at 4 and 8 weeks after TAC.
Results:Compared with Sham-operation group, the cardiac function and structure had obvious changes in the other 2 groups. Compared with TAC group, the ejection fraction in EP+ TAC group increased 15%, the heart weight index and left ventricular weight index decrease 15.7%and 20%respectively at 8 weeks after TAC, all P<0.05. Compared with Sham-operation group, the mRNA and protein expressions of ANP and BNP increased in TAC group at 4 and 8 weeks after TAC, increased in EP+TAC group at 8 week after TAC. Compared with TAC group, the mRNA expressions of ANP and BNP in EP+TAC group decreased 47%and 62%at 4 weeks after TAC, decreased 44%and 28.1%at 8 weeks after TAC, all P<0.05;the protein expression of ANP and BNP in EP+TAC group decreased 22.3%and 48%at 4 weeks after TAC, decreased 21.5%and 38.3%at 8 weeks after TAC, all P<0.01.
Conclusion: EP may improve cardiac pathological hypertrophy in pressure over-loaded rats at the early stage, and delay the heart failure process.
10.Phosphorylcholine coating enhances biocompatibility of expanded polytetrafluoroethylene used in polymeric prosthetic heart valves
Ben ZHANG ; Dejun GONG ; Xiwu ZHANG ; Tongyi XU ; Lin HAN ; Hao TANG ; Fanglin LU ; Zhiyun XU
Chinese Journal of Tissue Engineering Research 2014;(34):5509-5514
BACKGROUND:Our preliminary study found that the monocusp valves made of ultramicropore expanded
polytetrafluoroethylene (ePTFE) revealed no significant thrombus, calcification, or degradation 20 weeks after implanted into the descending aorta and the left pulmonary artery in sheep, which verified the good property of ePTFE. However, the surface of ePTFE in the left pulmonary artery was covered with obvious neointima.
OBJECTIVE: To assess the biocompatibility of phosphorylcholine-coated ePTFE.
METHODS:ePTFE surface was modified by phosphorylcholine derivative. Then the changes of surface shape, tensile stress at yield and elasticity modulus, water contact angle, and protein absorption capacity of ePTFE after surface modification were observed. (1) Hemolytic test: the leaching solution of phosphorylcholine-coated ePTFE, leaching solution of uncoated ePTFE, normal saline, and distiled water were added to the diluted human blood, respectively. (2) Platelet count test: the phosphorylcholine-coated ePTFE, uncoated ePTFE, high density
polyethylene, and Zymosan A were added to the whole blood samples from healthy volunteers, respectively.
(3) Platelet activation test: the phosphorylcholine-coated ePTFE, uncoated ePTFE, γ-Globulins, and Zymosan A
were added to the whole blood samples from healthy volunteers, respectively.
RESULTS AND CONCLUSION: The mean micropore diameter of ePTFE was significantly decreased after
phosphorylcholine coating (P < 0.001). The hydrophilicity and the ability of suppressing protein adsorption were
significantly strengthened after phosphorylcholine coating (P < 0.001). Phosphorylcholine coating did not influence
ePTFE in biomechanical properties and hemolytic test. The platelet count test and platelet activation test demonstrated that phosphorylcholine coating significantly improved anti-thrombus function of ePTFE. So, phosphorylcholine coating can enhance anti-thrombus function, suppress protein adsorption, and improve biocompatibility of ePTFE.