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。     

P27 Cyclin D_1在胆囊癌中的表达及意义

目的研究p2 7、CyelinDl 在胆囊癌中的表达情况 ,并探讨它们与肿瘤分化程度及生物学行为的关系。方法应用免疫组织化学S -P法检测 10例胆囊炎及 32例胆囊癌标本p2 7及CyelinDl的表达状况。结果p2 7在胆囊炎上皮中为低表达 ,在胆囊癌中少部分表现出高表达。而CyelinDl 在胆囊炎上皮中低表达 ,而在胆囊癌中大部分呈现高表达 ,两者间差异显著 (P <0 .0 5 )。结论CyclinDl在胆囊癌的发生发展中有重要作用 ,且与患者预后有关。     

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.     

P38MAPK抑制剂对缺血/再灌注大鼠肾脏功能损伤的保护作用

目的：观察P38MAPK抑制剂对缺血／再灌注大鼠肾脏功能损伤的保护作用。方法：夹闭肾动脉制遣大鼠肾脏缺血／再灌注损伤动物模型，静脉注射P38MAPK抑制剂阻断P38MAPK信号转导通路，测量其对肾功能及细胞因子含量的影响。结果：肾脏和血浆中TNF—α和IL-β含量随缺血／再灌注时间的延长而升高，肾功能损伤也随缺血／再灌注时间的延长而加重。应用P38MAPK抑制剂可显著降低TNF-α和IL-β含量，对缺血／再灌注所致的肾功能损伤具有明显改善作用。结论：P38MAPK抑制剂可通过抑制致炎因子的产生而减轻缺血/再灌注所致的大鼠肾脏功能损伤。     

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。     

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.     

Suppression of Gl Arrest and Enhancement of G2 Arrest by Inhibitors of Poly(ADP-ribose) Polymerase: Possible Involvement of Poly(ADP-ribosyl)ation in Cell Cycle Arrest Following γ-Irradiation

Low-dose γ-irradiation of mouse embryonic fibroblast C3D2F1 3T3-a cells caused Gl arrest along with G2 arrest and inhibition of replicative DNA synthesis. When the cells were cultured in the presence of inhibitors of poly(ADP-ribose) polymerase [EC 2.4.2.30], such as 3-aminobenzamide, benzamide and luminol, Gl arrest of C3D2F1 3T3-a cells was suppressed and enhancement of G2 arrest was observed. In contrast, 3-aminobenzoic acid, a non-inhibitory analog of 3-aminobenzamide, did not suppress Gl arrest following γ-irradiation. These results suggest that the poly(ADP-ribosyDation reaction is critical for the pathway of Gl arrest and is also involved in the pathway of G2 arrest.     

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.     

Transplacental passage of mifepristone and its influence on maternal and fetal steroid concentrations in the second trimester of pregnancy

The maternal and fetal endocrine effects of the maternal administration of the anti-progestin mifepristone in mid-pregnancy have been investigated. Mifepristone and the metabolite RU 42,633 were detected in the fetal circulation and in the amniotic fluid 4, 24 and 48 h after oral ingestion. Maximum fetal plasma concentrations of mifepristone occurred 4 h after treatment indicating rapid placental transfer of the drug. No significant changes in progesterone, cortisol, oestradiol or aldosterone concentrations were detected in the maternal circulation after mifepristone treatment. No significant changes occurred in the fetal progesterone, oestradiol or cortisol concentrations, but a significant increase in fetal aldosterone occurred 4 and 24 h after treatment. The significance of these results is discussed in relation to the possible therapeutic uses of mifepristone for inducing labour.