紫外线致大鼠角膜上皮细胞凋亡的研究

目的：探讨紫外线对角膜的损伤机制中是否有细胞亡的存在。方法：用400μw/cm^2的紫外线照射大鼠角膜,取下后迅速固定,作石蜡切片,然后用原位末端标记技术（ISEL）检测凋亡的细胞。结果：照射5min组与对照组无明显差异（P＞0．05）,而照射15min组与对照组及照射5min组均有明显差异（P＜0．001）。结论：紫外线照射大鼠角膜,经过一定的潜伏期后,可以诱导其上皮细胞凋亡。     

Programmed cell death in brains of HIV-1-positive AIDS and pre-AIDS patients

Neuropathological studies have revealed that the brains of HIV-1-infected AIDS patients show the typical encephalitis and, in addition, neuronal loss. More recently, this neuronal cell loss has been thought to take place via programmed cell death (apoptosis) which has been demonstrated by an in situ end labelling (ISEL) technique. In this study 54 brains of HIV-1-positive patients were investigated by the ISEL technique to investigate whether apoptosis is also present in the brains of patients at the asymptomatic stage. Of these, 10 patients suffered from HIV encephalitis (HIVE), 8 had AIDS without neuropathological disorders and 36 were HIV-1-positive pre-AIDS patients. Apoptotic cells were detected in 6 of the 10 HIVE, 1 of the 8 AIDS without central nervous system (CNS) disease and 4 of the 36 asymptomatic individuals. A difference seen between the AIDS and pre-AIDS cases was that, in the latter, apoptotic cells were found in the white matter in all 4 cases, while only 2 of these 4 showed apoptotic neurons. The presence of apoptotic cells in a number, albeit small, of brains of HIV-1-positive pre-AIDS individuals, combined with abnormalities described previously in the same group of patients gives further support to the opinion that brain damage already occurs during the early stages of HIV infection. Received: 19 June 1995 / Revised: 31 July 1995 / Accepted: 6 September 1995     

Fas mediates apoptosis in steroid-induced myopathy of rats.

Recently, apoptotic cell death has been reported in differentiated skeletal muscle, where apoptosis was generally assumed not to occur. To investigate whether apoptosis may contribute to the steroid-induced myopathy, rats treated with triamcinolone acetonide (TA) for 9 days were sacrificed for detecting apoptosis by in situ end-labelling (ISEL) and DNA electrophoresis in soleus muscles. Immunohistochemical stainings of Fas antigen and p53 protein were performed to examine whether apoptosis-related proteins were present in the myopathy. Muscle fibre necrosis and apoptotic myonuclei appeared in soleus muscles following administration of TA, while control muscles showed no evidences for apoptosis. Fas antigen was not detected in control muscles, but expressed in soleus muscles of steroid-induced myopathy. Some of the Fas antigen-expressing muscle fibres were positive for ISEL. p53 Protein was not detected in any muscle fibres. These findings indicate that TA can induce apoptosis in differentiated skeletal muscles, and Fas antigen might be partly related to apoptotic muscle death in steroid-induced myopathy.     

骨髓增生异常综合征细胞凋亡及增殖研究

为探讨骨髓增生异常综合征（ＭＤＳ）病人骨髓内造血细胞无效性血生成的原因,对５例不同类型的ＭＤＳ患者的塑料包埋骨髓活检标本,用抗ＩＵｄＲ／ＢｒｄＵ单抗检测了ＭＤＳ患者骨髓细胞的标记指数。用原位末端标记（ＩＳＥＬ）技术和琼脂糖凝胶电泳检测ＤＮＡ梯状片段的方法研究了本组病例的骨髓细胞凋亡情况,并建立了ＩＳＥＬ和ＩＵｄＲ／ＢｒｄＵ双标记法,同时测定骨髓同一细胞的增殖和凋亡现象。结果表明５例ＭＤＳ患者的骨髓     

细胞凋亡在胆囊肿瘤发生发展中的作用

为探讨细胞凋亡在胆囊肿瘤形成中的作用，我们用原位末端标记法对２４例胆囊癌和１６例胆囊腺瘤中的凋亡细胞进行测定。结果显示：胆囊腺瘤中的细胞凋亡数低于原发性胆囊癌（Ｐ〈０．０１），高分化及Ｎｅｖｉｎ分期Ｉ、Ⅱ、Ⅲ期腺癌中的凋亡细胞数低于低分化及Ｎｅｖｉｎ分期Ⅳ、Ⅴ期者（Ｐ〈０．０５）。因此作者认为细胞凋亡在胆囊癌发病中起重要作用，可以作为预测胆囊癌病程和预后的指标。     

Effect of photodynamic therapy (PDT) on the expression of pro-apoptotic protein Bak in nasopharyngeal carcinoma (NPC)

BACKGROUND AND OBJECTIVE: The stimulatory effects of low intensity laser therapy (LILT) have been widely published for the treatment of chronic ulceration. In contrast to this previous work, the current study investigated its potential efficacy (by using a dosage of 9 J/cm2) in the management of acute wounds. For this purpose, uncomplicated postoperative wounds after minor podiatric surgery were examined. STUDY DESIGN/MATERIALS AND METHODS: The study was designed as a controlled group study. Ethical approval was granted by the University of Ulster's Research Ethics Committee. Patients (n = 9) presenting with a total of 12 wounds after minor surgical procedures (partial/total nail avulsions/electrosurgery) were recruited from the Podiatry Teaching Clinic, Northern Ireland. Patients attended the clinic once per week for assessment and treatment. Weekly irradiation was performed by using a CBM Master 3 (CB Medico, Copenhagen, Denmark) diode laser (GaAlAs). The physical parameters of the output of this unit were as follows: wavelength, 830 nm; average power output, 30 mW; spot size, 0.1 cm2; irradiance, 300 mW/cm2; continuous wave output. Wound assessment and recording of pain levels were conducted weekly. Wound measurement was completed by using planimetry and digitising methods. RESULTS: Current findings indicated no statistically significant differences between Laser and Control groups for wound closure (P = 0.28 digitising; P = 0.49 planimetry) nor for pain levels reported (P = 0.88). CONCLUSION: It would seem that LILT provides no advantages in the management of minor postoperative wounds over current practice. Despite no apparent benefit of infrared laser at this dosage in the management of acute stage wounds, further research is required to determine its potential efficacy in the management of other wound types.     

Apoptotic response of spermatogenic cells to the germ cell mutagens etoposide,adriamycin, and diepoxybutane

In testis, apoptosis is a way to eliminate damaged germ cells during their development. In this study, we evaluated the ability of three germ cell mutagens to induce apoptosis (or programmed cell death) at specific stages of rat seminiferous epithelial cycle. These chemicals include the cancer chemotherapy drugs etoposide and adriamycin and the butadiene metabolite diepoxybutane. According to our results, etoposide is a very potent inducer of apoptosis in male rat germ cells and the cell types most sensitive to it include all types of spermatogonia, zygotene, and early pachytene spermatocytes and meiotically dividing spermatocytes. Also, adriamycin causes an increase in apoptosis at specific stages of seminiferous epithelial cycle and the most sensitive cell types are type A_3–4 spermatogonia, preleptotene, zygotene, and early pachytene spermatocytes. Die poxybutane does not cause any significant increase in the frequency of apoptosis in rat testis. In addition, we studied whether p53 is taking part in the apoptotic response of spermatogenic cells by studying the levels of p53 protein in testis before and after chemical treatment. No accumulation of p53 in testis was seen after treatment with these three chemicals. The expression of two p53-regulated genes, p21^WAF1 and mdm2, was also studied but no increase in the levels of mRNA of these genes was observed after treatment. The results indicate that apoptosis should be taken into consideration when the genotoxic effects of chemicals are evaluated in germ cells. Environ. Mol. Mutagen. 31:133–148, 1998 © 1998 Wiley-Liss, Inc.     

PCD检测方法的建立及其在研究外来激素对大鼠睾丸细胞PCD影响的应用

本研究1.建立了检测PCD的石蜡切片原位末端标记( ISEL ) 法, 在石蜡切片原位用免疫组化法标记核DNA,使 PCD 细胞呈黄色或棕黄色的阳性着染, 在普通光学显微镜下即可鉴别。本实验用加热和蛋白酶K同时处理背景, 使非特异性染色几乎不存在; 使用过氧化物酶—显色系统缩短时间2小时,提高了筛检DAB 速度。2.应用ISEL法研究外来激素对大鼠睾丸细胞的影响,结果发现GnRH-A是一个促凋原,具有明显的生殖毒性。     

Increased cell apoptosis in bone marrow trephine biopsies and immunomagnetically isolated myeloid progenitor cells in patients with chronic idiopathic neutropenia

The frequency of apoptotic cells in bone marrow trephine biopsies and cytospins of immunomagnetically isolated myeloid progenitor cells was determined in 39 patients with chronic idiopathic neutropenia (CIN) and 12 hematologically normal individuals using the in situ end-labeling (ISEL) apoptosis detection method. We found that 66.7% of the patients but none of the normal controls displayed apoptotic cells equal to or higher than 5% of the total mononuclear cells in bone marrow biopsies (p<0.01). In the double stain, we also found that the proportion of apoptotic CD15⁺ myeloid precursor cells did not differ significantly between patients and control subjects, while the proportion of apoptotic CD34⁺ hemopoietic cells could not be estimated with accuracy because of the presence of CD34⁺ endothelial cells. Significantly increased apoptosis was noted in cytospins of immunomagnetically isolated patient CD34⁺ and CD34⁺/CD33⁺ cells but not CD34^-/CD33⁺ cells, compared to the controls (p<0.001, p<0.02 and p>0.05, respectively). These findings confirm and extend our previous observations in flow-cytometric studies of apoptosis in CIN, indicating that increased apoptosis in CIN bone marrow concerns mainly the CD34⁺ and CD34⁺/CD33⁺ progenitor cell compartments. We conclude that the accelerated apoptosis in these compartments may account for the impaired neutrophil production in CIN patients.