Regional difference in the appearance of apoptotic cell death in the ligamentum flavum of the human cervical spine

Ossification or calcification of the ligamentum flavum (LF) is relatively common in the middle and lower cervical, thoracic, and lumbar spine but extremely rare in the upper cervical region. This clinical fact suggests that there exist local factors promoting or preventing ossification or calcification of LF. However, little is known about the differences in the ultrastructure and cellular alterations of the LF between the different spinal levels, even in the cervical spine. With electron microscopy, we examined samples of LF collected surgically from the upper and lower cervical spine regions; we then studied the apoptotic appearance of ligament cells using a preferential labeling method. We found direct evidence of apoptosis of ligament cells in the LF. Apoptosis was more apparent in the upper region samples than in the lower region samples. The spaces around the normal fibroblasts were filled with thick collagen fibrils, but the collagen fibrils disappeared around the apoptotic bodies and thin fibrils were formed. The difference of the level of apoptosis may correlate to the ultrastructual difference of LF, and our data will benefit further investigations seeking to clarify the mechanism of various pathological conditions in the human LF.     

Human multiple myeloma cells express peroxisome proliferator-activated receptor gamma and undergo apoptosis upon exposure to PPARgamma ligands

Multiple myeloma is essentially an incurable malignancy and it is therefore of great interest to develop new therapeutic approaches. We previously reported that human B cell-lymphomas express the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) and are killed by PPARgamma ligands. Herein, we investigate the therapeutic potential of PPARgamma ligands for multiple myeloma. The human multiple myeloma cell lines ANBL6 and 8226 express PPARgamma mRNA and protein. The PPARgamma ligands, 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) and ciglitazone, induced multiple myeloma cell apoptosis as determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, loss of mitochondrial membrane potential, and caspase activation. Importantly, the ability of PPARgamma ligands to kill both multiple myeloma cell lines was not abrogated by Interleukin-6 (IL-6), a multiple myeloma growth survival factor. Finally, the RXR ligand 9-cis retinoic acid (9-cis RA) in combination with PPARgamma ligands greatly enhanced multiple myeloma cell killing. These new findings support that PPARgamma ligands may represent a novel therapy for multiple myeloma.     

Immunohistochemical characteristics of duodenal adenomas in familial adenomatous polyposis with special reference to cell kinetics

The duodenum is the second most frequent site of cancer in patients with familial adenomatous polyposis (FAP). The main objective of this study was to evaluate the cell kinetics in duodenal and ampullary adenomas in FAP. The endoscopic and biopsy findings of duodenal adenomas in 22 FAP subjects and 18 non-FAP subjects were compared. Adenomas in FAP included 15 ampullary adenomas and 17 nonampullary adenomas. The cell kinetics was evaluated by immunohistochemistry for Ki-67, p53, bcl-2, and cyclooxygenase 2 (COX2), and the apoptotic index (AI) as determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method. Any correlations between the indices for cell kinetics and the endoscopic findings were identified. All 50 adenomas were histologically verified to be tubular adenoma with low-grade dysplasia. Neither the expression of Ki-67, p53, bcl-2, and COX2 nor the AI differed substantially between FAP and non-FAP subjects. In patients with FAP, duodenal adenoma tended to have a higher Ki-67-labeling index than the ampullary adenoma (54.3 +/- 11.3 versus 46.8 +/- 12.7; .05 < P < .1). In addition, the Ki-67-labeling index in endoscopically normal or slightly enlarged ampullary adenoma was significantly higher than that in markedly enlarged ampullary adenoma (51.8 +/- 11.4 versus 39.4 +/- 11.3; P < .05). Duodenal adenoma in FAP subjects was not found to have a higher proliferative activity or a smaller degree of apoptosis compared with those in non-FAP subjects. The smaller proliferative activity in larger ampullary adenoma may thus be related to the static nature of ampullary adenoma in FAP.     

豚鼠耳蜗外毛细胞凋亡时听力变化的实验观察

目的观察耳蜗外毛细胞发生凋亡时听力改变情况.方法对20只豚鼠药物造模,诱发耳蜗外毛细胞发凋亡.应用TUNEL技术观察凋亡表达,测试ABR阈值观察听力变化.结果应用丁胺卡那霉素1天即可诱发豚鼠耳蜗外毛细胞发生凋亡,连续应用3d,耳蜗外毛细胞凋亡呈强阳性表达,但ABR阈值无明显改变;随着用药时间延长,凋亡细胞数目增加,甚至出现部分毛细胞缺失现黎,此时ABR阈值明显升高.结论耳蜗外毛细胞发生凋亡早期对豚鼠听力无明显影响,随着耳毒性药物应用时间延长,豚鼠ABR阈值升高可能存在两种原因毛细胞凋亡或毛细胞坏死.     

Studies on hepatocyte apoptosis,proliferation and oncogene c-fos expression in carbon tetrachloride-induced cirrhotic rat liver

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保尔佳对原发性肝癌患者sIL-2R、IL-6、TNFα的影响及促进肝癌细胞凋亡的初步研究

对２０例原发性肝癌患者（大剂量组１０例、小剂量组１０例）临床应用保尔佳１周，用ＥＬＩＳＡ法检测治疗组用药前后外周血及用药后门静脉血的ｓＩＬ－２Ｒ，ＩＬ－６，ＴＮＦα的水平变化及用末端转移酶标记法（ＴＵＮＥＬ法）检测治疗组肝癌细胞的凋亡情况，与未治疗组、正常对照组相比。结果表明，治疗组外周血ｓＩＬ－２Ｒ水平明显下降（大剂量组Ｐ＜０．０１、小剂量组Ｐ＜０．０５），ＩＬ－６水平明显上升（大剂量组Ｐ＜０．０１、小剂量组Ｐ＜０．０５），ＴＮＦα水平有所下降（大剂量组Ｐ＜０．０５、小剂量组Ｐ＞０．０５），治疗组门静脉血与外周血ｓＩＬ－２Ｒ、ＩＬ－６、ＴＮＦα两者间无明显差异（Ｐ＞０．０５）。ＴＵＮＥＬ法检测显示保尔佳能促进肝癌细胞的凋亡（Ｐ＜０．０１，大、小剂量组间Ｐ＞０．０５）。且保尔佳对肝癌患者的血常规、肝功能等无不良影响。保尔佳是一种具有增强机体免疫功能、促进肿瘤细胞凋亡且无明显毒副作用的新型抗癌药物。     

Blocking tumor necrosis factor-alpha inhibits folic acid-induced acute renal failure

Systemic administration of mice with folic acid (FA) has been used for studying the pathogenesis of acute renal failure. However, the molecular mechanisms by which FA induces acute renal failure remain poorly understood. We found that CD-1 mice treated with FA developed acute renal failure characterized by increased blood urea nitrogen, necrosis, and apoptosis of tubular epithelial cells. Compared to control mice, tumor necrosis factor-alpha (TNF-alpha) was markedly elevated in blood and kidneys of these FA-treated mice, accompanied by markedly reduced expression of anti-apoptotic protein BclxL in their kidneys. In vivo administration of FA-treated CD-1 mice with neutralizing anti-TNF-alpha antibody restored the expression of BclxL in kidneys and inhibited the necrosis and apoptosis of renal tubular epithelial cells, leading to the amelioration of acute renal failure. In ex vivo cultures, we found that FA enhanced production of TNF-alpha, decreased expression of BclxL protein, and induced apoptosis of mouse cortical tubule (MCT) cells. Addition of neutralizing anti-TNF-alpha antibody, but not control IgG, in the cultures markedly blocked the apoptotic death of FA-treated MCT cells and restored expression of BclxL to the same levels as those MCT cells cultured in the absence of FA. All these results suggest that TNF-alpha is a critical inflammatory cytokine responsible for FA-mediated acute renal failure. Furthermore, in vivo administration of anti-TNF-alpha antibody may be proved as an effective approach for acute renal failure prevention and treatment.     

Epithelial Apoptosis and Loss in Airways of Children with Asthma

Objective. To examine loss and apoptosis of bronchial epithelial cells in children with asthma. Methods. We examined endobronchial biopsies from 13 asthmatic children and 11 non-asthmatic control subjects with other respiratory diseases. Postmortem samples were obtained from six children who died from non-respiratory diseases. We examined bronchial epithelial shedding by morphology; expression of caspase-3 and terminal deoxynucleotidyl-mediated dUTP nick end labeling (TUNEL) were used to study bronchial epithelial apoptosis. Results. We found epithelial loss to be increased in asthmatic children compared with non-asthmatic control subjects (p = .001) and postmortem children (p = .001). Caspase-3⁺ epithelial cells were significantly greater in children with asthma compared with both non-asthmatic control subjects (p = .001) and the postmortem group (p = .002); TUNEL⁺ epithelial cells were also increased in columnar cells in the asthmatic children compared with the non-asthmatic control subjects (p = .002) and the postmortem group (p = .001). Eosinophilia was absent in 11 of 13 asthmatic children, although they tended to have submucosal lymphocyte infiltration. Smooth muscle and mucus gland hyperplasia were seen in some asthmatic children whose biopsy specimens included these structures. Basement membranes of childhood asthmatics were thicker than in non-asthmatic controls (p = .002) and postmortem subjects (p = .001). Conclusion. Generally, apoptosis and loss of bronchial epithelial cells were increased in childhood asthma; increased apoptosis might be related to epithelial loss.     

电针对大鼠脊髓损伤后细胞凋亡相关基因影响的研究

目的：观察电针对大鼠急性脊髓损伤后神经细胞凋亡表达及变化规律。试图从线粒体启动凋亡途径相关主要因素的研究角度进一步揭示电针治疗脊髓损伤作用机制。方法：采用A1len’s法制备急性脊髓损伤模型。动物分为电针组、药物组（Caspase-3抑制剂组）、模型组、假手术组。用Tunel法对细胞凋亡进行标记。结果：脊髓损伤后1d即发现神经细胞凋亡,1d凋亡的细胞主要是神经元细胞。凋亡的细胞主要位于脊髓灰质前角。结论：大鼠急性脊髓损伤后存在细胞凋亡现象,是脊髓继发性损伤主要病理机制。