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.
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.
Objective To investigate the cell growth inhibition and apoptosis induced by rapamycin on human hepatocellular carcinoma Bel-7402 cells and to study the role of mitochondrium membrane potential in the process of apoptosis. Methods Bel-7402 cells in vitro were given 5, 10, 20, 30, 40 and 50 nmol/L different concentrations of rapamycin, and the cell growth inhibiting ratio of Bel-7402 was assessed by MTT assay. The changes of morphology of Bel-7402 were observed by Hoechst 33258 staining and flow cytometry (FCM), respectively; The cell mitochondrial membrane potential was detected by using JC-1 staining method. Results Rapamycin could inhibit the growth of Bel-7402 cells significantly by inducing apoptosis, and the growth suppression and the cell apoptosis both presented time-effect relationship and were also dose-dependent. The rates of inhibiting and cell apoptosis after 72 h exposure to 50 nmol/L rapamycin were significantly higher that those of other groups (P＜0.01). Typical morphological changes of cell apoptosis were observed very clearly after the Bel-7402 cells had been exposed to rapamycin for 48 hours using Hoechst 33258 staining method, and it was also observed that the mitochondrial membrane potential decreased when apoptosis occured (P＜0.01). Conclusion Rapamycin could inhibit the growth of Bel-7402 cells by inducing cell apoptosis, and the descent of mitochondrial membrane potential may play an important role in the process of cell apoptosis.
Objective To study the relationship between the expression of apoptosisrelated gene bclx,bax and estrogen receptor (ER) in primary gallbladder carcinoma (PGC) and its clinical significance. MethodsImmunohistochemistry of labeled dextran polymer (LDP) with EnvisionTM system was used to detect ER and gene bclx and bax. ResultsThe positive rate of bclx,bax and ER were 72.3%,66.0% and 59.6% in 47 cases with primary gallbladder carcinoma and 40.0%,93.3% and 93.3% in 6 cases with gallbladder adenomahyperplastic. The expression of bax and ER in PGC was significantly lower than that in gallbladder adenomahyperplastic (P＜0.05),the expression of bclx was significantly higher in PGC than that in the latter (P＜0.05).The expression of bclx and ER in well differentiated PGC was significantly higher than that in moderately, poorly differentiated PGC (P＜0.05); bax expression in well differentiated PGC was lower. ER and bax expression in male PGC was significantly lower than that in female PGC （P＜0.01）, the expression of bclx in male PGC was higher (P＜0.05).ER was more highly expressed in smaller PGC than in larger one (P＜0.05). ER and bax, bclx were not different between various clinical stages and ages (P＞0.05,respectively). Conclusion The expression ER, apoptosisrelated gene bclx and bax have correlation with differentiation and sex in PGC, their levels shows significance in the prognosis of PGC.
ObjectiveThis study is aimed to determine the expression of ubiquitin-specific peptidase 39 (USP39) protein in the colorectal cancer (CRC) tissues, and the effect of silencing USP39 gene on the cell growth and cell cycle distribution of CRC cells.Methods① The expressions of USP39 protein in CRC tissues and its paracancerous tissues were determined by immunohistochemical staining method. ② By lentiviral infection, Lv-shUSP39 (KD-1 and KD-2 group) and Lv-shCon (shCon group) were transferred into SW1116 and HCT116 cells, and cells of blank control group did not received any treatment (Con group). To determine the role of USP39 gene in cell growth, MTT assay was performed to draw growth curve, and cell cycle distribution of CRC cells in the 4 groups were determined by flow cytometer.Results① The expression of USP39 protein was higher in CRC tissues compared to adjacent tissues (P=0.007). ② For SW1116 and HCT116 cells, the cell proliferation ability of KD-1 and KD-2 groups were remarkably decreased than those in corresponding shCon and Con groups on 3, 4, and 5-day (P<0.05). ③ Flow cytometry assay showed that, the percentage of G0/G1 phase cells were decreased obviously (P<0.05), while increased significantly in percentage of G2/M phase and number of sub-G1 phase cells in KD-1 group compared with that in the Con group and shCon group of SW1116 and HCT116 cells (P<0.05).ConclusionsThe expression of USP39 protein is highly expressed in CRC tissues. Knockdowning of USP39 gene can inhibit cell proliferation and promote cell apoptosis.
Objective
To explore the effect of pyropheophorbide-a methyl ester-mediated photodynamic therapy (MPPa-PDT) on the apoptosis in human osteosarcoma cell line MG63 and the underlying mechanism.
Methods
Human osteosarcoma MG63 cells in logarithmic growth phase were divided into 4 groups: blank control group (control group), the MPPa treatment group (MPPa group), the light irradiation group (LED group), and MPPa-PDT treatment group (MPPa-PDT group). MPPa-PDT group and MPPa group were incubated with MPPa (0.75?μmol/ L) for 20 hours in dark condition; control group and LED group were incubated with equal volume of fresh medium for 20 hours in the same condition. After washing with PBS and replacement with fresh culture medium, LED group and MPPa-PDT group cells were exposed to light (4.8 J/cm2) for 120 seconds. After light exposure, all groups were cultured in dark condition again. Then cellular morphology changes were observed by an inverted phase contrast microscopy, endoplasmic reticulum morphology changes were observed by transmission electron microscopy, cellular apoptosis was detected by Hoechst33258 nuclear staining, cell apoptotic rate and the levels of Ca in cells were analyzed by flow cytometry, the expression of p-PERK, C/EBP homologous protein (CHOP), cleaved-Caspase-12 were assayed by Western blot.
Results
In MPPa-PDT group, the retracted and round cells were observed; Hoechst33258 nuclear staining showed nuclear condensation, fragmentation, and other typical apoptotic morphological changes; the cell apoptotic rate (48.76%±3.54%) was significantly higher than that of control group (5.04%±0.41%), MPPa group (5.33%±0.38%), and LED group (6.48%±0.46%) (P < 0.05); the levels of Ca2+ in cells (485.29±58.77) was also significantly higher than that of control group (97.24±4.77), MPPa group (97.95±6.30), and LED group (101.17±5.26) (P < 0.05); swelling endoplasmic reticulum was observed under transmission electron microscope; the expressions of p-PERK, CHOP, and cleaved-Caspase-12 gradually increased at 1, 3, and 6 hours after treatment respectively, which were significantly higher than those of the other groups (P < 0.05). There was no typical apoptotic morphological changes and endoplasmic reticulum morphological changes in control group, MPPa group, and LED group, and there was no significant difference in the above indexes among 3 groups (P > 0.05).
Conclusion
MPPa-PDT can significantly induce apoptosis in MG63 cells. The endoplasmic reticulum stress pathway is involved in the MPPa-PDT induced apoptosis.
