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OBJECTIVE: To observe the effect of acupuncture of intraorbital and extraorbital acupoints on apoptosis and expression of Bax, Bcl-2 and Caspase-3 proteins of retinal ganglion tissue in rabbits with nonarteritis anterior ischemic optic neuropathy (NAION), so as to reveal its mechanism underlying improvement of NAION. METHODS: Female New Zealand rabbits were randomly divided into four groups: model, intraocular needling (ION), extraocular needling (EON), ION＋EON groups (n＝5 per group), and the 5 healthy eyes of those rabbits in the model group were selected to be used as the control group. The NAION model of the right eye was established by photodynamic stroke method. For ION group, acupoints "Jingming" (BL1), "Chengqi" (ST1) and "Qiuhou" (EX-HN7) were needled, and for EON group, "Cuanzhu" (BL2), "Yuyao" (EX-HN4) and "Qiaoming" (EX) were needled with filiform needles, followed by retaining the needles for 30 min. For ION＋EON group, the 6 acupoints were needled simutaneously. The treatment was conducted once daily for 3 days. The apoptosis of retinal ganglion tissue was detected by using TUNEL fluorescence labeling, and the expression of Bax, Bcl-2 and active Caspase-3 in the retinal ganglion were detected by immunohistochemistry. RESULTS: Following modeling and compared with the control group, the number of the apoptotic cells, and the expression levels of Bax and Caspase-3 proteins, as well as the ratio of Bax/Bcl-2 were significantly increased in the model group (P0.05). CONCLUSION: Acupuncture of intraorbital and extraorbital acupoints can reduce apoptosis of retinal ganglion in NAION rabbits via inhibiting the activation of Caspase-3 protein and ratio of Bax/Bcl-2, and up-regulating the expression of Bcl-2 protein.
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Background & objectives: Significance of apoptosis as a prognostic marker is less well studied in paediatric acute lymphoblastic leukaemia (ALL) cases. Hence, a prospective study, involving 30 paediatric ALL cases, was done to assess the clinical relevance of in vivo apoptosis. Methods: Peripheral blood mononuclear cells from all patients were subjected to annexin V/propidium iodide staining to detect the degree of apoptosis [apoptotic index (AI)] at day 0 and day 35 post-induction chemotherapy. In addition, Bax and Bcl2 apoptotic protein expressions were studied at day 0 and their relative fluorescence mean intensity (RFMI) ratios were calculated. Results: Mean age of patients was 5.1 years. Of the 30 cases, 21 (70%) were at standard-risk, five (17%) at intermediate and four (13%) at high risk. Majority (83%) were B-ALL. Day 8 absolute blast count was >1000/?l in seven (23%) and <1000/?l in 23 of 30 (77%) cases. Day 35 marrow was M1 in 23 (92%) and M2 in two of 25 (8%) cases. AI at day 0 and day 35 ranged from 0.9 to16.6 per cent and 1.4 to 62.8 per cent with a mean of 5.90 and 19.64 per cent, respectively. The Bax/Bcl2 ratio ranged from 0.2 to 3.5 with a mean of 0.83. The ratio was predominantly anti-apoptotic, i.e. <1 (77%). A significant association was noted between low AI at day 0 and high total leucocyte count (P=0.02), T-cell phenotype (P=0.043) and high-risk as per NCI category (P=0.025). Significant increase (>30%) in day 35 AI was seen in only six cases. Interpretation & conclusions: Our study showed that low AI at day 0 was associated with a high-risk clinical phenotype in paediatric ALL. However, studies on larger group, especially with longer follow up or study of relapse cases, will help draw conclusions regarding apoptosis assessment in paediatric ALL.
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The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.