SB203580对肾缺血/再灌注损伤时细胞凋亡及p38MAPK影响的实验研究

目的　探讨不同时间及不同浓度p38MAPK特异性抑制剂SB2 0 35 80对肾缺血 /再灌注损伤过程中肾功能、细胞凋亡及 p38MAPK活性、表达量、p38MAPK底物的影响。 方法　 4 9只大鼠按缺血 /再灌注及给药时间的不同 ,随机分为 7组 ,每组7只大鼠按正交拉丁方表的顺序 ,经尾静脉注射相同体积、不同剂量的SB ,使其在大鼠体内达到不同的浓度。测定BUN和Scr;用TUNEL试剂盒检测细胞凋亡情况 ;Westernblot技术用于蛋白定性及半定量分析。结果　SB可显著减轻大鼠肾缺血 /再灌注损伤造成的Scr和BUN的升高、肾小管上皮细胞的凋亡及 p38MAPK的激活 ,但存在模型、剂量及给药时机的差异 (P<0 .0 5 ) ,缺血前 3h之前给药 ,使其体内血药浓度达到 5 μmol/L左右可取得较好的效果。 结论　SB可显著减轻大鼠肾缺血 /再灌注损伤 ,在缺血前 3h之前给药 ,同时使其血药浓度达到 5 μmol/L左右可取得最佳效果。     

胶质细胞瘤复发前后Cyclin D1和P16的表达

目的　探讨CyclinD1,P16在胶质瘤复发前后表达改变及其意义。方法　采用免疫组织化学LsABC法对 4 5例复发胶质瘤瘤组织、瘤旁脑组织和 10例正常脑组织CyclinD1,P16蛋白表达进行检测 ,统计分析CyclinD1,P16表达水平与胶质瘤分级、肿瘤复发的关系。结果　正常脑组织 ,瘤旁脑组织和胶质瘤组织CyclinD1表达依次升高 ,而P16的表达依次下降 ;肿瘤复发CyclinD1表达增强 ,P16的表达减弱。结论　CyclinD1与P16的表达与胶质瘤恶性进程和复发密切相关。     

Effect of N-tosyl-L-phenylalanylchloromethyl Ketone on Tumor Necrosis Factor-alpha -induced NF-κB Activation and Apoptosis in U937 Cell Line

Summary: To investigate the effect of N-tosyl-L-phenylalanylchloromethyl ketone (TPCK) on tumor necrosis factor-alpha-induced NF-κB activation and apoptosis in U937 cell line, changes and subcellular localization of NF-κB/p65 and IκB-α were observed by fluorescencemicroscopy and expression and degradation of IκB-α by flow cytometry. The apoptosis of U937 cells was measured by flow cytometry and electrophoresis of DNA. Immunolfluorescence assay showed that NF-κB/p65,IκB-α only localized in cytoplasm. After TNF-α stimulation, p65 was localized only in nuclei, and IκB-α was only localized in cytoplasm and decreased. The changes of TNF-α stimulation were specifically inhibited by TPCK. Flow cytometry also revealed the downregulation of IκB-α protein during TNF-α-induced apoptosis and the down-regulation was specifically inhibited by TPCK. Flow cytometry also showed the apoptosis of U937 cells after TNF-α induction. DNA ladder can be detected in cells treated by TNF-α. It is concluded that degradation of IκB-α protein and NF-κB/p65 translocation occur during TNF-α-induced apoptosis of U937 cells, suggesting the activation of NF-κB.TPCK-sensitive protease plays an important role in the degradation of IκB-α protein induced by TNF-α in U937 cells. TPCK sensitive protease also plays an important role in the apoptosis of U937 cells induced by TNF-α.     

皮肤鳞状细胞癌中p53、bcl-2和c-myc的表达及意义

目的 研究p53、bcl—2和c—myc在皮肤鳞状细胞癌中的异常表达及其与癌分化程度的关系。方法 应用免疫组织化学S-P法检测58例皮肤鳞状细胞癌中p53、bcl—2和c—myc的表达水平。结果 58例皮肤鳞状细胞癌中p53、bcl-2和c—myc的阳性表达率分别为48．3％、67．2％和32．8％。结论 p53、bcl—2和c—myc异常表达在皮肤鳞状细胞癌的发生、发展中起重要作用，并与皮肤癌的恶性程度有关。     

Association of a 70-kDa tyrosine phosphoprotein with the CD16:ζ:γ complex expressed in human natural killer cells

The CD16: ζ: γ receptor complex allows natural killer (NK) cells to recognize and eliminate antibody-coated target cells. Whereas the ectodomain of CD16 is the receptor for Fcγ domains of immunoglobulins, disulfide-linked homo- and heterodimers composed of ζ and γ are required for the cell surface expression, and signal transduction properties of the complex. Engagement of CD16 activates the tyrosine kinase pathway, which induces the tyrosine phosphorylation of several substrates, including the ζ subunit and the phospholipase C γ-1 and γ-2 isoforms. Here we show that CD 16 stimulation of either peripheral blood NK cells, leukemic NK cells, or Jurkat transformants expressing a CD16:ζ:γ receptor complex, results in the tyrosine phosphorylation of a 70 kDa ζ-associated protein (pp70). Similarly, a 70-kDa ζ-associated phosphoprotein in T cells has been shown to be a tyrosine kinase (ZAP-70). Peptide mapping analysis indicates that the 70-kDa ζ-associated phosphoproteins from T cells and NK cells are structurally indistinguishable. We conclude that the CD16:ζ:γ complex may use a ZAP-70-related non-receptor tyrosine kinase, in the CD16 signaling cascade leading to NK cell activation.     

Effects of decentralization on the levels of GAP-43 and p38 (synaptophysin) in sympathetic adrenergic neurons: a semi-quantitative study using immunofluorescence and confocal laser scanning microscopy

The distribution of GAP-43 in superior cervical ganglion (SCG) and iris were studied in normal animals and following decentralization using immunofluorescence and confocal laser scanning microscopy (CLSM). GAP-43-like immunoreactivity (LI) was compared with p38 (synaptophysin)-LI, and tyrosin hydroxylase (TH)-LI. In the control SCG, GAP-43-LI and p38-LI were mainly localized in nerve terminals around the principal neurons. The neuronal perikarya were negative for GAP-43, but positive for p38 in a perinuclear zone, as well as positive for TH. SIF cells (Small Intensely Fluorescent cells, ganglionic interneurons) were positive for GAP-43, TH and p38. One day after decentralization, GAP-43-LI and p38-LI in nerve terminals around principal neurons had disappeared. Some of the principal neurons showed a weak GAP-43-immunoreactivity. Three days post-decentralization, GAP-43- and p38-positive nerve terminals around the neurons had reappeared in considerable numbers and the intra-ganglionic nerve bundles were positive for both antibodies. In the control irides, GAP-43-LI and p38-LI were distributed in a varicose pattern in the nerve bundles, around blood vessels and in the network of terminals. Double labelling studies showed that GAP-43-LI was colocalized with TH-LI and p38-LI. The network of terminals in the dilator plate of the irides was quantified by measuring the fluorescence intensity of randomly selected areas, using CLSM. Three days after decentralization the intensity of GAP-43-LI and p38-LI had significantly increased. TH-LI had decreased 8 days after decentralization. The results indicate that GAP-43-LI and p38-LI are normally present in the nerve fibers and terminals of both pre- and post-ganglionic neurons in adult rats. The expression of GAP-43-LI and p38-LI in post-ganglionic neurons is preganglionically regulated, as indicated by the increased expression after decentralization. The expression of p38 in these neurons is probably regulated via mechanisms that are separate from those which regulate GAP-43, since it showed a different time course than that of GAP-43-LI.     

雷公藤生物碱的分离鉴定

我们从雷公藤（ＴｒｉｐｔｅｒｙｇｉｕｍＷｉｌｆｏｒｄｉｉＨｏｏｋ．ｆ．）中按常法分离得到总碱后经ＨＰＬＣ分析出现８个峰，经硅胶柱层析及硅胶ＲＰ－１８反相中压柱层析分离得到５个生物碱，由化学方法和光谱分析证明为雷公藤吉碱（Ｗｉｌｆｏｒｇｉｎｅ２０）．雷公藤次碱（Ｗｉｌｆｏｒｉｎｅ２１），雷公藤春碱（Ｗｉｌｆｏｒｔｒｉｎｅ２２），雷公藤碱戊（Ｗｉｌｆｏｒｉｄｉｎｅ２３），呋喃南蛇碱（Ｃｅｌａｆｕｒｉｎｅ１６）。ＨＰＬＣ分析指出含量最高为雷公藤吉碱，其次为呋喃南蛇碱。     

Correlation of Nuclear Morphometry and Immunostaining for p53 and Proliferating Cell Nuclear Antigen in Transitional Cell Carcinoma of the Bladder

Background :
In an attempt to determine the biological significance of nuclear morphometric findings, measurements of mean nuclear volume (MNV) and nuclear roundness factor (NRF) were compared to the immunoreactivityof p53 expression and proliferating cell nuclear antigen (PCNA) in human bladder cancer.
Methods :
MNV and NRF were measured using stereological methods. Expression of p53 and PCNA were determined by immunohistochemical staining. Specimens from 111 patients with previously untreated bladder cancer were analyzed.
Results :
The mean MNV was 235.8 ± 1 33.6 μm³ for the 81 patients with p53-labeling index (LI) less than 10% and 337.2 ± 141.0 μn³ for the 30 patients with p53 LI greater than 10% (P = 0.008). There was Resign if icant correlation between NRF and expression of p53. The mean MNV was 220.1 ± 1 20.5 μm³ for the 67 patients with PCNA LI less than 28% (the mean value of PCNA LI) and 328.9 ± 149.2 μm³ in 44 patients with PCNA LI greater than 28% (P= 0.0001). The mean NRF was 80.7 ± 4.2 for the 67 patients with PCNA LI less than 28%, and 82.3 ± 3.4 for the 44 patients with PCNA LI more than 28% (P= 0.04). Conclusion: Nuclear morphometric findings may reflect the proliferative potential of cancer eel Is of the bladder, as indicated by findings of immunostaining for p53 and PCNA.