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.
目的研究依達拉奉影響肝臟缺血再灌注過程中TNF-α的表達情況，探討依達拉奉對肝臟缺血再灌注損傷的逆轉作用。 方法將80只Wistar大鼠編號，根據計算機產生隨機數字，前40為一組，后40為一組，分為實驗組和對照組2組，建立常溫下部分肝缺血再灌注損傷動物模型。 在肝臟缺血再灌注損傷開始前1 h和開始時對實驗組大鼠給予依達拉奉注射液10 ml，對照組則給予同等容量的生理鹽水。分別于再灌注后0、1、2及4 h測定肝臟脂質過氧化物酶（LPO）和肝臟谷草轉氨酶(AST) 濃度； 應用RT-PCR法檢測肝組織TNF-α mRNA含量，并測定肝組織和血清中TNF-α水平； 應用TUNEL染色法檢測缺血肝組織的細胞凋亡情況。結果再灌注后1、2及4 h，實驗組大鼠肝臟LPO及AST濃度均明顯低于對照組（Plt;0.001）； 實驗組再灌注后1 h時肝組織TNF-α mRNA表達量、肝組織和血清TNF-α含量均明顯升高且達峰值，但均明顯低于對照組（Plt;0.05）； 再灌注后各時相實驗組肝細胞凋亡率明顯升高，但均明顯低于對照組（Plt;0.05）。 結論依達拉奉能抑制氧化應激反應，從而降低肝缺血再灌注損傷； 并顯著減少炎性細胞因子TNF-α的產生，抑制炎性反應的發生，減少肝細胞的凋亡。
ObjectiveTo observe apoptosis and proliferation of choledochus wall epithelial cell and fibrocyte, to understand the effects of apoptosis and proliferation on choledochal cyst development.MethodsThirty two cases of cystic dilatation,35 cases of cylindrical dilatation，and 25 cases of cholangiectasis caused by choledocholith were collected. All specimens were offered by department of hepatobiliarypediatric surgery. The apoptosis related index (bcl2 and bax) and cell proliferation index (PCNA) were detected by the immunohistochemical technique; Apoptosis was detected by TUNEL method. ResultsThere was serious mucosal epithelial cell damage in cystic dilatation group. In cylindrical dilatation group there was a damage similar to that of the cystis dilatation group, but the damage was not serious. In control group there was little damage in the duct wall, but there was a low positive rate of apoptosis of 〔epithelium cell (2.74±1.00)% and fibroblast (2.95±0.87）%〕, and a low bcl2 and bax’s expression rate, and a high PCNA’s expression rate 〔epithelium cell (3.74±1.00)%, fibroblast (3.71±1.77)%〕. There was no obvious difference between cylindrical dilatation group and cystic dilatation group (Pgt;0.05): the PCNA’s expression rate was low 〔(0.99±0.51)% and (0.90±0.38)% respectively〕, the bax expression rate was high in remaining epithelial cell, and the positive rate of bax was apparently higher than that of bcl2 (P＜0.05), the positive rate of the apoptosis cell was high 〔(13.94±4.77)%, (7.51±3.46)%〕; the expression rate PCNA were high 〔(9.91±2.91)%,(9.70±3.18)%〕, and expression rate of bax’s was low in the fibre tissue, the positive rate of bcl2 was markedly higher than that of bax, and the positive rate of the apoptosis cell was low 〔(3.74±2.12)%,(4.46±2.41)%〕. There were no marked difference between the two groups (Pgt;0.05). The expression of bcl2 and bax had marked difference both in cylindrical dilatation group and cystic dilatation group and as compared to control group (P＜0.05). ConclusionApoptosis has certain promoting effect in the course of choledochal cyst formation.
Abstract： Stem cell paracrine has been considered as the main mechanism to promote infarcted myocardium regeneration and repair of damaged cardiomyocytes. With further research, secreted frizzled-related protein 2 （Sfrp 2） and stem cell paracrine are closely linked to each other. Sfrp 2 can competitively bind to the specific receptor Fz in Wnt signaling pathway, inhibit Wnt signaling pathway to regulates apoptosis, differentiation, and other life processes of stem cells, and therefore becomes a research hotspot in recent years. This review focuses on the mechanism of Sfrp 2/Wnt signal way in stem cell therapy for myocardial infarction.
目的：了解大鼠腦出血后血腫周圍組織細胞凋亡與神經元特異性烯醇化酶（NSE）的表達及大鼠神經功能缺損程度的關系。方法：用膠原酶注入到大鼠尾狀核的方法制作腦出血模型。將大鼠分為腦出血、假手術組、正常組3組。采用蘇木素伊紅(HE) 染色、NSE免疫組織化學染色及TUNEL分別觀察各組在腦出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d時血腫周圍NSE及TUNEL的表達。用Longa評分法評價大鼠神經功能缺損程度。結果：大鼠在膠原酶注入6 h后形成穩定的血腫，在造模24～48 h神經功能缺損程度最重；6 h即見到TUNEL陽性細胞的表達，在48 h最明顯；NSE從神經元中漏出彌散到細胞間隙也在48 h達高峰。結論：腦出血血腫周圍凋亡與神經功能缺損及NSE的變化有關，凋亡可能在腦出血的神經損傷中起重要的作用。
摘要:目的:動態觀察大鼠腦出血后血腫周圍組織補體激活與細胞凋亡的規律。方法:用膠原酶注入到大鼠尾狀核的方法制作腦出血模型。將大鼠分為腦出血、假手術組、正常組3組。采用蘇木素伊紅(HE) 染色、免疫組織化學染色及原位末端脫氧核苷酸轉移酶介導的dUTP 缺口末端標記法(TUNEL)分別觀察各組在腦出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d時血腫周圍補體C3、促凋亡基因(Bax)、抑凋亡基因(Bclxl)及TUNEL的表達。結果補體C3的表達峰值在24~48 h;TUNEL、Bax蛋白表達術后12h增加,48~72 h達高峰,而Bclxl蛋白表達高峰在48h。結論:大鼠腦出血后血腫周圍組織補體C3的表達增加與細胞凋亡的演變趨勢一致,C3與凋亡有相關。Abstract: Objective: To study the complement activation and apoptosis regular genes changes in the tissues of the perihematoma of intracerebral hemorrhage (ICH) in rats. Methods: Intracerebral hemorrhage was induced in rats by injection of bacterial collagenase into the caudate nucleus. Histopathological changes were studied in 6 h,12 h, 24 h, 2 d, 3 d, 5 d, 7 d after the injection. The immunohistochemistry and TUNEL analysis were performed. The expression of complement factor C3, the TUNELpositive cells, the proapoptotic gene expression (Bax) and the antiapoptotic gene (Bclxl) were examined. Results: The expression of C3 increased to its maximum between 2448 h. The TUNELpositive cells and Bax protein expression increased gradually and reached the peak level between 4872 h. The Bclxl protein reached the peak level at 48 h. The correlation analysis showed that the quantity of C3 was positively related to that of the TUNELpositive cells, but the bax protein was not related to Bclxl protein. Conclusion: The expression of complement factor C3 may contributes to the nerve injury after cerebral hemorrhage and relate to the apotosis in the tissues surrounding the hametoma in rats.
ObjectiveTo investigate the effect of electroacupuncture on the apoptosis of hippocampal neurons in C57BL/6J mice with status epilepticus by observing the changes of hippocampal subtle neuron pathology and apoptosis.MethodsMale C57BL/6J mice were used to prepare epileptic status models of lithium-pilocarpine mice, and then 7-day electroacupuncture stimulation (Baihui, Fengfu) were given to the mice model. Open field experiment and new object recognition experiment were performed to observe the changes of cognitive abilities. The pathological changes of hippocampal neurons were detected by HE staining. Hippocampal apoptosis protein (Caspase-3) and microtubule-associated protein (MAP-2) were detected by immunohistochemistry. Effect of electroacupuncture on apoptosis of hippocampal neurons in C57BL/6J mice with status epilepticus were recorded.Results① Compared with the control group, the vertical movement, modification times, and number of crossings of the model group all decreased significantly (P<0.000 1,P<0.000 1,P<0.000 1), and their cognitive ability decreased significantly (P<0.01). Compared with the model group, vertical movements, modification times, and number of crossings were increased in the electroacupuncture (EA) group (P<0.01,P<0.05,P<0.05), and the cognitive ability of new objects was increased (P<0.01). ② HE staining showed that the model group had significant damage to the hippocampal neurons of mice, and the cells swelled, nuclear collapsed and vacuoles appeared. In the EA group, the injury of hippocampal neurons was alleviated, and cell edema and vacuolization were alleviated. ③ Immunohistochemistry showed that compared with the control group, the IOD of the Caspase-3 positive cells in the hippocampus of the model group increased significantly (P<0.000 1), and the IOD of the MAP-2 positive cells decreased significantly (P<0.01); Compared with the electroacupuncture, the IOD of the Caspase-3 positive cells in the hippocampus of the mice decreased (P<0.05), and the IOD of the MAP-2 positive cells increased (P<0.05).ConclusionsElectroacupuncture can improve the pathological changes of hippocampal neurons in C57BL/6J mice with status epilepticus, promote cytoskeletal repair, reduce neuronal apoptosis in hippocampus, and antagonize the damage of hippocampal neurons induced by status epilepticus.
Expression of bcl-2 in 45 cases of primary gallbladder carcinomas and 39 cases of gallbladder adenomas were detected by streptavidin-biotin-peroxidase complex (SABC) methods. The results revealed that the positive rate of bcl-2 in primary gallbladder carcinomas was 71.1% and that in gallbladder adenomas was 87.2%, there was no obvious difference between them (Ｐ＞0.05). Expression of bcl-2 in well, moderately and poorly differentiated primary gallbladder carcinomas were 82.8%, 82.4% and 50.0% respectively with significant statistical difference (Ｐ＜0.01). The positive rate of bcl-2 in stages Ⅰ,Ⅱ and Ⅲ of primary gallbladder carcinomas were 81.3% and 55.2% respectively with no significant differnce. The result suggests that expression of bcl-2 in primary gallbladder carcinomas has no correlation with clinic stages.
Objective To investigate the regulatory effect of somatostatin analogue (SMS201995,SMS) on proliferation and apoptosis in human cholangiocarcinoma cell line in vitro. MethodsProliferation curve, flow cytometry, agarose gel electrophoresis, Annexin VFITC and flow cytometric immunofluorescent technique were performed to identify the inhibitory effect on cell proliferation and the induction of apoptosis of human cholangiocarcinoma cells (SKChA1). ResultsSMS significantly reduced the SKChA1 cell growth by serum in long experiments and transiently accumulated it in G0/G1 phase. Dotplot analysis of cells duallabeled with Annexin VFITC and PI confirmed the induction of apoptosis by SMS in SKChA1 cells.AnnexinVFITC labeling was markedly enhanced following treatment with SMS for 24 h. DNA of treated SKChA1 cells appeared a ladder pattern characteristic of apoptosis. Besides, timedependent increase in bax and decrease in bcl2 occured during SMS treatment. Conclusion SMS could inhibit the proliferation activity and induce apoptosis of cholangiocarcinoma cell line SKChA1. The mechanisms of apoptosis might be correlated with the expression of apoptosisregulatory gene bax and bcl2.