Objective To investigate the protective effects of antitumor necrosis factor-α antibody （TNF-αAb） on lung injury after cardiopulmonary bypass （CPB） and their mechanisms. Methods Forty healthy New Zealand white rabbits，weighting 2.0-2.5 kg，male or female，were randomly divided into 4 groups with 10 rabbits in each group. In groupⅠ，the rabbits received CPB and pulmonary arterial perfusion. In group Ⅱ，the rabbits received CPB and pulmonary arterial perfusion with TNF-αAb. In group Ⅲ，the rabbits received CPB only. In group Ⅳ，the rabbits only received sham surgery. Neutrophils count，TNF-α and malondialdehyde （MDA） concentrations of the blood samples from the left and right atrium as well as oxygenation index were examined before and after CPB in the 4 groups. Pathological and ultrastructural changes of the lung tissues were observed under light and electron microscopes. Lung water content，TNF-α mRNA and apoptoticindex of the lung tissues were measured at different time points. Results Compared with group Ⅳ，after CPB，the rabbitsin group Ⅰ to group Ⅲ showed significantly higher blood levels of neutrophils count，TNF-α and MDA（P＜0.05），higherTNF-α mRNA expression，apoptosis index and water content of the lung tissues （P＜0.05），and significantly lower oxyg-enation index （P＜0.05） as well as considerable pathomorphological changes in the lung tissues. Compared with group Ⅱ，after CPB，the rabbits in groups Ⅰ and Ⅲ had significantly higher blood concentrations of TNF-α （5 minutes after aortic declamping，220.43±16.44 pg/ml vs.185.27±11.78 pg/ml，P＜0.05；249.99±14.09 pg/ml vs.185.27±11.78 pg/ml，P＜0.05），significantly higher apoptosis index （at the time of CPB termination，60.7‰±13.09‰ vs. 37.9‰±7.78‰，P＜0.05；59.6‰±7.74‰ vs. 37.9‰±7.78‰，P＜0.05），significantly higher blood levels of neutrophils count and MDA （P＜0.05），significantly higher TNF-α mRNA expression and water content of the lung tissues （P＜0.05），and significantly loweroxygenation index （P＜0.05） as well as considerable pathomorphological changes in the lung tissues. Compared with groupⅠ，rabbits in group Ⅲ had significantly higher above parameters （P＜0.05） but lower oxygenation index （P＜0.05） only at 30 minutes after the start of CPB. Conclusion Pulmonary artery perfusion with TNF-αAb can significantly attenuate inflammatory lung injury and apoptosis of the lung tissues during CPB.
Objective To investigate the pathogenesis of acute lung injury in rats induced by intra-peritoneally injection of perforative peritonitis ascitic fluids（PPAF） and the role of L-arginine （L-Arg） in acute lung injury in this model. Methods Perforative peritonitis （PP） models were established in 60 rats and PPAF were collected. Forty-eight rats were randomly divided equally into NS group，PPAF group, and L-Arg group. Rats were randomly subjected to death at 7 h and 12 h. Peripheral blood WBC were counted，levels of NO and malondialdehyde （MDA） in serum were examined. Lung injury score and wet/dry ratio were evaluated， and level of myeloperoxidase （MPO） in lung tissues and lung cell apoptosis were tested. Results WBC count of peripheral blood, levels of NO and MDA in serum， level of MPO in lung tissue, lung injury score， wet/dry ratio， and lung cell apoptosis rate in PPAF group were significantly higher than that in NS group at each time point（P＜0.01）. Level of NO in serum in L-Arg group was higher than that in PPAF group （P＜0.01）， but lower level of MDA in serum， lower level of MPO in lung tissue and lung injury score，lower wet/dry ratio， and lung cell apoptosis rate were observed in L-Arg group（P＜0.05）. In PPAF group and L-Arg group， level of NO in serum， wet/dry ratio, and lung cell apoptosis rate were higher at 12 h than that at 7 h（P=0.000）. Serum NO level was in negative correlation with serum MDA level （r=-0.257，P=0.021）， MPO level in lung tissue（r=-0.444， P=0.011），and lung cell apoptosis（r=-0.351， P =0.010） in PPAF group and L-Arg group， but serum MDA level was in positive correlation with cell apoptosis（r=0.969， P＜0.001） in each group. Conclusions Acute lung injury rats model can be established by intra-peritoneally injection of PPAF. Enhanced oxidizing reaction and cell apoptosis take part in the occurrence of acute lung injury. L-Arg plays a protective role in acute lung injury.
ObjectiveTo investigate the possible mechanism of cucurmosin on apoptosis in human pancreatic cancer cell line SW1990 in vitro. MethodsThe inhibition of cucurmosin on SW1990 cell was detected by MTT assay, the apoptosis was observed by transmission electron microscope, the apoptosis rate was analyzed by flow cytometry, and the protein level of caspase3 was determined by Western blot. ResultsAfter exposure to cucurmosin at 1.25, 2.50, 5.00, 10.00, 20.00, 40.00, and 80.00 μg/ml for 24, 48, and 72 h, the proliferation of SW1990 cell was inhibited in a time-and dose-dependent manner (Plt;0.05). At 72 h after 40.00 μg/ml cucurmosin treatment, the typical apoptosis changes and apoptotic bodies were observed by transmission electron microscope. After exposure to cucurmosin at 0, 2.50, 10.00, and 40.00 μg/ml for 72 h, the apoptosis rate increased gradually as (0.30±0.11)%, (18.93±1.06)%, (28.00±2.07)%, and (49.93±3.25)%, respectively (Plt;0.05). The expression of caspase-3 protein was elevated gradually (Plt;0.05). ConclusionCucurmosin may induce the apoptosis of pancreatic cancer cell through up-regulating the expression of caspase-3.
OBJECTIVE: To explore the effect of Fas/Apo-1 and Bcl-2 gene expression on mechanism of scar formation. METHODS: Immunohistochemical method was applied to defect the expression of Fas and Bcl-2 protein in fibroblasts from 10 cases with normal skin, 10 cases with hypertrophic scar and 10 cases with keloid. RESULTS: The positive expression rate of Bcl-2 protein in keloid was 83.2%, significantly higher than that in hypertrophic scar (38.6%), (P lt; 0.01), and the positive expression rate in hypertrophic scar and keloid was higher than that in normal skin (6.78%), (P lt; 0.01). But the positive expression rate of Fas/Apo-1 protein was 78.4% in normal skin 80.4% in hypertrophic scar, 84.4% in keloid respectively, which showed no significant difference among them (P gt; 0.05). CONCLUSION: Bcl-2 gene but Fas gene may take part in the formation of pathologic scar.
Objective To investigate whether the agonist of delta opoid receptor D-Ala(2),D-Leu(5) enkephalin (DADLE) has the effect of decreasing myocardial injury during ischemia-reperfusion of adult rabbits’ myocardium，so that a new mehanism and way to myocardial protection could be found. Methods Langendorff model was used during the experiment. Thirty rabbits were divided into three groups randomly (each group 10 rabbits). Control group: St.Thomas Ⅱ cardioplegic solution was used; group 1: St.Thomas Ⅱ cardioplegic solution and DADLE (1mg/kg) were used; group 2: St.Thomas Ⅱ cardioplegic solution and naloxone(3mg/kg) were used to induce the hearts to arrest respectively. After arrest the hearts were reperfused respectively. Data of left ventricle development pressure(LVDP) was recorded before and after ischemia. Biochemical indicators of myocardium, lactate dehydrogenase(LDH) were detected before and after ischemia. Some myocardial tissues were used to explore the changes of the tissue of ultrastructure with electron microscope，when the experiment was over. Still some myocardial tissues were to be detected by flow cytometer to evaluate the apoptosis of the myocardium. Results The LDH and LVDP showed significant difference among three groups after ischemia(Plt;0.05); LVDP in group 1 was higher than those in group 2 and control group(69.8±5.8 mmHg vs. 23.4±3.9 mmHg; 69.8±5.8 mmHg vs. 37.9±4.7 mmHg; Plt;0.05), the LDH in group 1 was lower than those in group 2 and control group(1 272.6±59.1 U/L vs. 2 764.4±27.7 U/L, 1 272.6±59.1 U/L vs. 1 884.4±37.5 U/L; Plt;0.05). The apoptosis rate in group 1 was lower than those in group 2 and control group. As could be shown from the ultrastructure: mitochondria structure was nearly normal in group 1; mitochondria structure was injuried severely in group 2; there was a minor injury in control group. Conclusion Agonist of δ opoid receptor DADLE in cardioplegic solution could induce hibernation, which has myocardial protection effect during ischemia-reperfusion injury.
Objective To evaluate the phenomena of apoptosis and its relevant mechanism during ischemia-reperfusion period. Methods The published papers to explore the apoptotic phenomena and its mechanism in organs or tissues which experienced ischemia-reperfusion injury were reviewed. Results Apoptosis was common in ischemia-reperfusioned organ or tissue. The severity of apoptosis was influenced by many factors such as ischemia, hypoxia, oxygen free radials, intracellular free calcium ion overloading, various cytokines, et al; and also was regulated by bcl-2 family, caspase family and NF-κB，et al. Conclusion Apoptosis is a common phenomenum in ischemiareperfusioned organ or tissue which is affected and regulated by various factors.
Abstract: Objective To investigate the influence of cardiopul monary bypass(CPB) to the cellular immune function of T lymphocyte. Me th ods Among 500 patients operated from March 2006 to September 2006，30 patients with rheumatic heart disease were selected randomly as the CPB group, which would replace mitral valve; 30 patients with congenital patent ductus arte reriosus as the nonCPB group, which would ligate ductus arteriosus without CPB . The blood was sampled before operation, at the end of CPB or operation, and 24 hours after operation. After T lymphocyte was seperated, the quantum o f T lymphocyte, apoptosis of T lymphocyte, ability of T lymphocyte to kill tumou r cell were measured. Results The quantum of T lymphocyte i n CPB group at the end of CPB was decreased than that before operation （50.9% ±6.8% vs. 58.5%± 9.1%,Plt;0.05）; apoptosis of T lymphocyte at the end of CPB and 24 hou rs after operation were increased than that before operation （6.5%±2.2% vs. 0. 9%±1.1%, 5.6%±1.8% vs. 0.9%±1.1%;Plt;0.01）; ability to kill tumour cell b reakdown in CPB group at the end of CPB and 24 hours after operation was decrea sed than that before operation （30.4%±6.0% vs. 37.3%±8.6%, 29.0%±4.9% vs . 37 .3%±8.6%;Plt;0.05）. Ability to kill tumour cell breakdown in CPB group was lower than that in nonCPB group at the end of CPB （30.4%±6.0% vs. 33.6%±5. 3%, Plt;0.05）. Conclusion CPB can depress the cellular im mune function,which causes temporary immune depression to the body.
目的:探討丙型肝炎病毒非結構蛋白NS4B在原發性肝癌發生中的作用,及其發生機制。方法:設置對照組、空白載體PCXN2組、轉染NS4B組。使用脂質體介導法,轉染丙型肝炎病毒非結構蛋白重組質粒NS4B進入Chang肝細胞內,并用G418篩選。繪制生長曲線,分別用流式細胞儀檢測瞬時表達及穩定表達時肝細胞凋亡率,用均數±標準差表示,統計學分析采用Dunnett t檢驗及q檢驗。結果:瞬時表達各組(PCXN2,NS4B)凋亡率分別為:(918±060)%,(445±053)%。穩定表達各組(PCXN2,NS4B)凋亡率分別為:(1575±209)%,(366±034)%。與PCXN2組比較Plt;001。結論:NS4B抑制肝細胞凋亡率,可能導致肝細胞異常增殖,誘導肝癌發生。
Objective To investigate the role of chemokine receptor CXCR7 in the development and progression of pancreatic carcinoma. Methods The short hairpin RNA (shRNA) targeting CXCR7 was designed and delivered into AsPC-1 pancreatic carcinoma cells to knock down CXCR7 expression. The cell proliferation, cell cycle, and apoptosis after CXCR7 knockdown was determined by MTT and flow cytometry, respectively. The invasive ability of pancreatic carcinoma cells was evaluated by using the Transwell system. Results Compared with the blank control group (BC group), transfection of AsPC-1 cells with CXCR7-shRNA resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P<0.05), and the ability of proliferation and invasion significantly decreased (P<0.05). Knockdown of CXCR7 also significantly increase apoptosis (P<0.05), induce cell cycle arrest at G0/G1 phase (P<0.05). Conclusions Taken together, the present study showes that the knockdown of CXCR7 expression may play an important role in pancreatic carcinoma development, invasion, and metastasis, CXCR7 may be a potential therapeutic target for the treatment of pancreatic carcinoma.
Objective To observe the influences of uncoupling protein 2 (UCP-2) rs660339 variants transfection on cell proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods Two UCP-2 green fluorescent protein (GFP) lentivirus constructs were created with the rs660339 locus carried C or T (UCP-2C or UCP-2T), respectively. HUVEC were cultured after lentiviral infection of UCP-2C or UCP-2T. The expression of UCP-2C or UCP-2T was detected with real time polymerase chain reaction. Cell proliferation and cell apoptosis were compared among negative control (NC) group, UCP-2T group and UCP-2C group using CCK-8 cell viability and flow cytometry. Western blot and immunostaining were employed to examine the expression of Bcl-2 gene. Results The lentivirus constructs were successfully created. ＞80% of the transfected cells were found to express GFP under fluorescent microscope. The mRNA levels of UCP-2 gene were significantly increased (F=29.183,P=0.001) in the UCP-2T group and UCP-2C group. The CCK-8 assay revealed that on day two (F=15.970,P=0.004), day three (F=16.738,P=0.004), day four (F=5.414,P=0.045) post-infection, UCP-2T and UCP-2C group showed significantly greater proliferation than the NC cells. The apoptotic rate in the UCP-2T and UCP-2C group was significantly lower than NC group (F=277.138,P=0.000), and the apoptotic rate of UCP-2T was significantly lower than that of UCP-2C (P=0.003). The protein levels of Bcl-2 in the UCP-2T and UCP-2C group were significantly greater than that in the NC group (F=425.679,P=0.000), and the Bcl-2 expression of UCP-2T was greater than that of UCP-2C (P=0.002). The Bcl-2 density in the UCP-2T and UCP-2C group were greater than that in the NC group (F=11.827,P=0.008), while there was no difference between UCP-2T and UCP-2C group (P=0.404). Conclusion The variants of UCP-2 rs660339 may influence HUVEC proliferation and apoptosis, and UCP-2T showed a stronger effect of inhibiting apoptosis than UCP-2C.