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：了解大鼠脑出血后血肿周围组织细胞凋亡与神经元特异性烯醇化酶（NSE）的表达及大鼠神经功能缺损程度的关系。方法：用胶原酶注入到大鼠尾状核的方法制作脑出血模型。将大鼠分为脑出血、假手术组、正常组3组。采用苏木素伊红(HE) 染色、NSE免疫组织化学染色及TUNEL分别观察各组在脑出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d时血肿周围NSE及TUNEL的表达。用Longa评分法评价大鼠神经功能缺损程度。结果：大鼠在胶原酶注入6 h后形成稳定的血肿，在造模24～48 h神经功能缺损程度最重；6 h即见到TUNEL阳性细胞的表达，在48 h最明显；NSE从神经元中漏出弥散到细胞间隙也在48 h达高峰。结论：脑出血血肿周围凋亡与神经功能缺损及NSE的变化有关，凋亡可能在脑出血的神经损伤中起重要的作用。
摘要:目的:动态观察大鼠脑出血后血肿周围组织补体激活与细胞凋亡的规律。方法:用胶原酶注入到大鼠尾状核的方法制作脑出血模型。将大鼠分为脑出血、假手术组、正常组3组。采用苏木素伊红(HE) 染色、免疫组织化学染色及原位末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记法(TUNEL)分别观察各组在脑出血后第6 h、12 h、24 h、48 h、72 h、5 d、7 d时血肿周围补体C3、促凋亡基因(Bax)、抑凋亡基因(Bclxl)及TUNEL的表达。结果补体C3的表达峰值在24~48 h;TUNEL、Bax蛋白表达术后12h增加,48~72 h达高峰,而Bclxl蛋白表达高峰在48h。结论:大鼠脑出血后血肿周围组织补体C3的表达增加与细胞凋亡的演变趋势一致,C3与凋亡有相关。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.
摘要:目的:探讨卡配因抑制剂3（MDL28170）对新生大鼠缺氧缺血性脑损伤(HIBD)神经细胞凋亡的影响。方法:建立新生SD大鼠HIBD模型,治疗组于缺养缺血后即刻、2 h、4 h腹腔内注射MDL28170,对照组及手术组同时予生理盐水。缺氧缺血后24 h用免疫组化方法观察大脑皮质及海马CA1区Caspase3 蛋白表达、TUNEL法检测细胞凋亡,观察组织病理改变并计算海马神经元死亡数,透射电镜观察细胞超微结构。结果:缺氧缺血后24 h缺血侧大脑皮质及海马CA1区Caspase3和TUNEL阳性细胞数较对照组明显增加,透射电镜证实有凋亡细胞;MDL28170可减少阳性细胞数量,抑制神经元死亡,差异有显著性(Plt;0.05)。结论:MDL28170可通过抑制神经凋亡而对新生大鼠HIBD具有一定保护作用。Abstract: Objective: To investigate the effect of (Calpain inhibitor3) MDL28170 on neural apoptosis in a neonatal model of hypoxicischemic brain damage (HIBD). Methods: A neonatal model of HIBD was established, 7dayold SD rats were divided into three groups. The treatment group received MDL28170(ip) at 0 h,2 h,4 h after HI, whereas the other two groups were administered normal saline simultaneously. The expression of caspase3 (by immunohistochemistry), neural apoptosis (by TUNEL) in cortex and hippocampus ipsilateral to the insult were observed 24 h after HI; hippocampal CA1 neural loss and electromicroscopic changes were assessed at the same time. Results: Apoptotic body was observed by electromicroscopy. Caspase3 positive cells and apoptotic cells increased significantly in the ipsilateral cortex and hippocampal CA1 region compared to the control, and MDL28170 reduced the number of positive cells, attenuated CA1 neural loss with significance (Plt;0.05). Conclusion: It is suggested that MDL28170 may protect the brain of neonatal rats after HIBD by suppressing neural apoptosis.
目的:通过检测髓母细胞瘤（Medulloblastoma）中凋亡抑制蛋白（survivin）的表达情况，探讨它的临床意义，为髓母细胞瘤的治疗寻求新的思路。方法:对50例髓母细胞瘤及10例正常脑组织石蜡切片行免疫组化检测，通过对survivin蛋白在髓母细胞瘤患者与正常脑组织中的表达差异，并运用相关统计分析方法分析它在不同性别、年龄段、肿瘤大小、病理类型、临床分期的表达差异及其与预后的相关性。结果:（1）髓母细胞瘤中survivin蛋白阳性表达率为72%，明显高于正常脑组织的阳性表达率0%，二者表达差异明显（Plt;0.01）；亚组分析发现在survivin蛋白阳性组（n=36）和阴性组（n=14）病例中，性别分布、肿瘤部位、肿瘤大小、病理分型等因素在两者间无明显差别（Pgt;0.05），而年龄及TM临床分期在两组间差异显著（P分别为0.044和0.027）。（2）survivin阳性患者预后与和阴性组相比差异显著（P=0.001）。结论:survivin可能参与了对髓母细胞瘤细胞凋亡的调控，并对髓母细胞瘤的发生发展可能起着重要作用。由于survivin蛋白表达与髓母细胞瘤患者年龄、临床分期及预后等也密切相关，可作为判断病情和评价预后的指标。
研究白介素6(IL-6)在过氧化氢诱导的肺泡上皮细胞凋亡中的作用及其机制。方法：以人Ⅱ型肺泡上皮细胞A549细胞株为实验对象，MTT法检测IL-6对A549细胞增殖活性的影响，TUNEL检测法检测IL-6对A549细胞凋亡率的影响，Western blot观察caspase-3和caspase-9蛋白量的变化。结果：IL-6能够增加A549细胞的增殖活性，减少其凋亡。与模型组相比，IL-6干预组的caspase-3和caspase-9的蛋白表达量明显降低,有统计学意义。结论：IL-6能够抑制过氧化氢诱导的肺泡上皮细胞凋亡，并且与caspase-3和caspase-9蛋白的表达量下降有关。提示IL-6对肺泡上皮细胞的保护作用可能是通过抑制细胞凋亡相关蛋白来实现的。
目的：观察青蒿琥酯对脂多糖诱导的大鼠肾小球系膜细胞(glomerular mesangial cells,GMC)凋亡的影响。方法：用不同浓度的青蒿琥酯刺激脂多糖诱导增殖的大鼠肾小球系膜细胞，HE染色观察凋亡细胞形态，DAPI荧光染色观察细胞核凋亡情况，用流式细胞仪检测细胞凋亡率。结果：青蒿琥酯对系膜细胞凋亡有明显诱导作用，呈一定剂量依赖性。结论：青蒿琥酯能够剂量依赖性地诱导大鼠系膜细胞凋亡。
Objective To explore repair role of allogeneic bone marrow mesenchymal stem cells （BM-MSCs） transplantation on treating hepatic ischemia reperfusion injury （HIRI） in rats. Methods Ten rats were executed to get BM-MSCs， then BM-MSCs were cultured in vitro and dyed by 4，6-diamidino-2-phenylindole （DAPI）. Models of 70% hepatic ischemia reperfusion injury were eatablished. Thirty two rats were randomly divided into sham operation group （Sham group）， ischemia reperfusion group （I/R group）， Vitamin C group （VC group）， and BM-MSCs group. Serum samples were analyzed for ALT and AST， and hepatic tissue were for superoxide dismutase （SOD） and malondialdehyde （MDA）. Liver sections were stain with hematoxylin and eosin （HE） for histological analysis， TUNEL staining was applied to detect hepatic apoptosis. Serum and tissues were both collected at 24 h after reperfusion. Results The isolated BM-MSCs maintained vigorous growth in vitro. Specific markers for MSCs antigens CD29 and CD44 were detected by flow cytometry， but antigens CD34 and CD45 were not be detected. Models of HIRI were stable， and BM-MSCs were detected around the periportal area by DAPI staining. Compared with I/R group， levels of ALT， AST， MDA， and AI in the VC group and BM-MSCs group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Compared with VC group， levels of ALT， AST， MDA， and AI in the BM-MSC group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Conclusion BM-MSCs could protect HIRI by alleviating oxidative stress and inhibiting cellular apoptosis.
Objective To review the relationship between the expression levels of bcl-2, bax and bad gene and other biological factors of breast cancer in the growth and development of breast cancer. Methods Related literatures were summarized and reviewed. Results The expression level change of antiapoptosis gene bcl-2 was still under research and the expression levels of apoptosis gene bax and bad were down-regulated progressively in the evolution from benign breast tissue to breast cancer tissue. The expression level of bcl-2 had positive correlation with some positive factors in breast cancer such as estrogen receptor （ER） and progesterone receptor (PR), while it had negative correlation with some negative factors such as p53, EGFR, c-erbB-2 and lymph node metastasis. The levels of ER, PR and the expression level of p53 of breast cancer had no relationship with the expression level of bax. Up to now there was no report about the relationship between the expression level of bad and other biological factors of breast cancer. Conclusion The role of altered expression level of bcl-2, in the treatment and prognosis of breast cancer is still controversial, and the relationship between the expression of bad and the prognosis of breast cancer is still unknown, but expression level of bax is correlated positively with the prognosis of breast cancer. Research on these genes can provide us some new index to evaluate the prognosis of breast cancer and new ideas on treatment of breast cancer including gene therapy.
Objective To study the effects of glutaminase (GA) gene blocked by antisense nucleotide on apoptosis of transplanted gastric carcinoma cells in nude mice. Methods The plasmid containing antisense sequence of GA gene was trans-fected into gastric carcinoma cells , then the cells were injected to endermic tissue of nude mice to create animal models of gastric carcinoma. Apoptosis of tumor cells was detected by terminal deoxynucleotidyl transferase2mediated nick end labeling (TUNEL) method. The expression of GA mRNA in tumor tissue was measured by reverse transcription polymerase chain reaction (RT2PCR) technique. Results After the successful transfection of plasmid containing antisense sequence of GA gene into gastric carcinoma cells , the tumor’s growth speed decreased , apoptosis of tumor cells increased , and the expression of GA mRNA also decreased. Conclusion The antisense gene of GA could inhibit the expression of GA gene and significantly increase the apoptosis of gastric carcinoma cells.