Immunohistochemical analysis of 14-3-3 sigma and related proteins in hyperplastic and neoplastic breast lesions, with particular reference to early carcinogenesis

In order to confirm the role of 14-3-3 sigma (sigma) as a tumor suppressor in breast carcinogenesis, we have studied the expression of 14-3-3sigma immunohistochemically in usual ductal hyperplasia (UDH), ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) breast lesions. Immunostaining for estrogen receptor alpha (ERalpha), p53 and estrogen-responsive RING finger protein (Efp) was also carried out. Immunohistochemically, expression of 14-3-3sigma was seen in 92% UDH lesions and gradually decreased from 65% in DCIS to 23% in IDC. The expression of ERalpha decreased gradually from UDH to DCIS to IDC, while p53 showed an inverse staining pattern to that of ERalpha. The expression of Efp showed no significant difference among the three breast lesions. Hence, the present immunohistochemical study confirmed 14-3-3sigma as a tumor suppressor in breast carcinogenesis. A similar immunohistochemical analysis was then carried out on columnar cell hyperplasia with atypia (CCHA), in which the expression pattern of tumor suppressor 14-3-3sigma, ERalpha and p53 suggested that it might be possible that CCHA is a precancerous lesion.     

术前碳水化合物对瘢痕切除术后胰岛素抵抗的影响

目的观察术前口服碳水化合物及静脉输注葡萄糖对术后胰岛素抵抗程度的影响.方法择期瘢痕切除术病人60例,随机分为对照组、口服组和静注组,每组20例.术前对照组常规禁食禁饮;口服组口服12.5%碳水化合物的饮料（CHO）;静注组持续静脉输注10%葡萄糖液.分别于口服或静注前、术后6 h采血,测定血糖、血清胰岛素以及红细胞胰岛素受体.结果处理前各组血糖、血清胰岛素、胰岛素敏感指数及红细胞胰岛素高亲和力受体、低亲和力受体无明显差异;术后6 h口服组、静注组的血糖明显低于对照组,与处理前无显著差别;术后6 h各组血清胰岛素无显著差别,均显著高于处理前;胰岛素敏感指数、红细胞低亲和力受体三组均较处理前显著降低,但口服组、静注组明显高于对照组;术后6 h口服组、静注组红细胞高亲和力受体无明显降低;口服组与静注组间的血糖、胰岛素、胰岛素敏感指数及红细胞胰岛素受体无明显差别.结论术前静脉输注葡萄糖可缓解术后胰岛素抵抗的程度,术前饮用碳水化合物具有同样的效果,是一种简单有效的治疗方法.     

鼠抗人OX40功能性单克隆抗体的研制及其生物学特性的鉴定

目的：研制功能性鼠抗人OX40单克隆抗体。方法：以转人OX40的转基因细胞L929-OX40为免疫原，常规免疫6—8周龄的雌性BALB／c小鼠；采用B淋巴细胞融合技术，将免疫小鼠脾脏细胞与SP2／0融合，以L929-OX40转基因细胞及PHA活化的T细胞为抗体筛选阳性细胞，经免疫荧光标记分析对杂交瘤进行反复筛选和多次的克隆化培养；采用快速定性试纸法及竞争抑制结合试验分析了该单抗的亚类及抗原识别位点；采用MTT法分析单抗在体外对T细胞的促增殖效应以及ELISA分析活化T细胞分泌的细胞因子。结果：获得1株持续、稳定分泌鼠抗人OX40单克隆抗体的杂交瘤细胞株(命名为7E11)，该单抗能特异性地识别人OX40分子和介导有效的共刺激信号，体外促进活化的T细胞增殖和细胞因子的分泌。结论：成功研制成一株能分泌功能性鼠抗人OX40单克隆抗体的杂交瘤，该抗体特异性地识别人OX40分子并具有在体外协同刺激T细胞的作用。     

OGD增加小鼠离体海马组织内cPKCβII和nPKCε的膜转位

目的:通过观察无糖低氧(OGD)刺激对小鼠海马脑片内蛋白激酶C(PKC)特定亚型膜转位水平(激活程度)的影响,进一步证实我们在整体低氧预适应小鼠模型上所获实验结果,并为离体海马脑片缺血/低氧预适应(I/HPC)模型建立及后续药物干预实验打下基础。方法:急性分离小鼠海马组织、制备400!μm厚度的脑片,并用无糖低氧(OGD)人工脑脊液(ACSF)模拟缺血/低氧刺激;应用SDS-PAGE和Westernbolt等生化技术,并结合GelDoc凝胶成像系统,半定量检测OGD刺激2、5、10、15和30min后海马脑片内cPKCβII和nPKCε的膜转位水平。结果:OGD刺激可增高海马脑片内cPKCβII和nPKCε的膜转位水平,且这种增高从刺激5min开始持续至30min均有显著差异(P<0.001,n=6)。结论:实验结果进一步证实cPKCβII和nPKCε可能参与脑缺血/低氧性预适应的形成过程,并提示OGD10min作为制备离体海马脑片I/HPC模型的预处理刺激较为合理。     

广州市1996—2005年管网末梢水卫生状况及对策分析

目的了解广州市管网末梢水的水质情况，为随时监测全市生活饮用水水质的动态变化提供信息并提出改进措施。方法对广州市1996—2005年管网末梢水水质监测资料进行统计，按照GB5749—85《生活饮用水卫生标准》和卫生部《生活饮用水卫生规范》（2001年）进行分析评价。结果共监测606份水样，合格155份，合格率为25．58％。不合格的项目主要有锰、游离余氯、细菌总数、色度、铁和耗氧量。在广州市疾病预防控制中心检测的项目中，锰的合格率仅为59．57％，游离余氯的合格率为76．73％，细菌总数的合格率为85．81％，色度的合格率为88．12％，铁的合格率为88．94％，耗氧量的合格率为96．37％。结论广州市管网末梢水的锰、游离余氯、细菌总数和铁等项目合格率较低，今后应加强饮水的氯化消毒、对管网的定期检修和冲洗、减少二次供水水箱的使用和设立管网水水质自动监测系统。     

IFN-γ对体外培养系统中IL-18诱导的肿瘤特异性CTL杀伤效应的影响

目的应用rhIL-18在体外培养系统(Coculture system in vitro,CCs)中诱导肿瘤特异性细胞毒性T淋巴细胞(Cytotoxic T Lymphocyte,CTL),探讨IFN-γ在rhIL-18诱导的肿瘤特异性CTL产生过程及杀伤效应中的作用.方法采用Stem SepTM免疫磁性细胞分离法分离人外周血NK细胞、T细胞及DCs细胞,流式细胞仪分析细胞表型,125Ⅰ-UdR标记的细胞毒实验检测杀伤活性,ELISA方法检测IFN-γ蛋白产生量,RT-PCR检测IFN-γ mRNA表达含量.结果在肿瘤抗原存在的条件下,IL-18在CCs中能够诱导并促进CTL介导的肿瘤特异性杀伤效应;IL-18能够在肿瘤抗原刺激的CCs中诱导IFN-γ mRNA的表达及IFN-γ蛋白的产生,并与IL-18的含量呈剂量依赖关系,在rhIL-18含量为100 ng时,培养上清中IFN-γ为4 410±210 pg/ml.但加入抗IFN-γ抗体对rhIL-18诱导的肿瘤特异性CTL产生过程无明显影响,不能抑制IL-18诱导的这种肿瘤特异性CTL的杀伤效应.结论IL-18能够在肿瘤抗原刺激的CCs中有效地诱导CTL介导的肿瘤特异性杀伤效应,并诱导IFN-γ的产生,但其诱导肿瘤特异性CTL的产生过程及杀伤效应与IFN-γ无关.     

A comparison of the potency of the oxime HLö-7 and currently used oximes (HI-6, pralidoxime, obidoxime) to reactivate nerve agent-inhibited rat brain acetylcholinesterase by in vitro methods

(1) The efficacy of the oxime HL?7 and currently used oximes (pralidoxime, obidoxime, HI-6) to reactivate acetylcholinesterase inhibited by various nerve agents (sarin, tabun, cyclosarin, VX) was tested by in vitro methods. (2) Both H oximes (HL?-7, HI-6) were found to be more efficacious reactivators of sarin and VX-inhibited acetylcholinesterase than pralidoxime and obidoxime. On the other hand, their potency to reactivate tabun-inhibited acetylcholinesterase is very low and does not reach the reactivating efficacy of obidoxime. In the case of cyclosarin, the oxime HI-6 was only found to be able to sufficiently reactivate cyclosarin-inhibited acetylcholinesterase in vitro. (3) Thus, the oxime HL?-7 does not seem to be more efficacious reactivator of nerve agent-inhibited acetylcholinesterase than HI-6 according to in vitro evaluation of their reactivation potency and, therefore, it is not more suitable to be introduced for antidotal treatment of nerve agent-exposed people than HI-6.     

Attenuation of intracavitary applicators in 192Ir-HDR brachytherapy

Unlike the penetrating monoenergetic 662 keV gamma rays emitted by 137Cs LDR sources, the spectrum of 192Ir used in HDR brachytherapy contains low-energy components. Since these are selectively absorbed by the high-atomic number materials of which intracavitary applicators are made, the traditional neglect of applicator attenuation can lead to appreciable dose errors. We investigated the attenuation effects of a uterine applicator, and of a set of commonly used vaginal cylinders. The uterine applicator consists of a stainless steel source guide tube with a wall thickness of 0.5 mm and a density of 8.02 g/cm3, whereas the vaginal cylinders consist of the same stainless steel tube plus concentric polysulfone cylinders with a radius of 1 or 2 cm and a density of 1.40 g/cm3. Monte Carlo simulations were performed to compute dose distributions for a bare 192Ir-HDR source, and for the same source located within the applicators. Relative measurements of applicator attenuation using ion-chambers (0.125 cm3) confirmed the Monte Carlo results within 0.5%. We found that the neglect of the applicator attenuation overestimates the dose along the transverse plane by up to 3.5%. At oblique angles, the longer photon path within applicators worsens the error. We defined attenuation-corrected radial dose and anisotropy functions, and applied them to a treatment having multiple dwell positions inside a vaginal cylinder.     

SK&F 95654-induced acute cardiovascular toxicity in Sprague-Dawley rats--histopathologic, electron microscopic, and immunohistochemical studies

The characteristics and pathogenesis of the cardiovascular toxicity induced by the type III selective phosphodiesterase inhibitor SK&F 95654 were examined in 2 studies. Sprague-Dawley rats received either a single sc injection of 50, 100, or 200 mg/kg SK&F 95654 and were euthanized at 24 hours after administration of the drug (Study 1), or were given a single subcutaneous (sc) injection of 100 mg/kg SK&F 95654 and euthanized at 1, 2, 4, 6, 8,12, 24 hours, or 2 weeks after treatment (Study 2). Control rats received either DMSO or saline. Myocardial lesions and vascular lesions of the mesentery, spleen, and pancreas were seen 24 hours after dosing with either 50,100, or 200 mg/kg SK&F 95654. The frequency and severity of these lesions (evaluated after the 100 mg/kg dose) increased with time over a period of 1 to 24 hours. By 2 weeks, the lesions subsided. Cardiac lesions consisted of myocyte necrosis with hypercontraction bands, inflammatory cell infiltration, interstitial hemorrhage, and interstitial edema. Vascular lesions of the mesentery were most prominent and consisted of vasodilatation and inflammation in the small-sized vessels, arterial medial necrosis and hemorrhage, and venous thrombosis. The vascular lesions included: leukocyte adhesion to endothelial cells, transendothelial migration of leukocytes, and inflammatory cell infiltration into vessel walls. Affected vessels included arteries, terminal arterioles, capillaries, postcapillary venules, and veins. Apoptosis of endothelial and smooth muscle cells was detected in the mesenteric vasculature by both TUNEL assay and electron microscopy. Evidence of endothelial cell activation in the mesenteric arteries and veins was also observed by electron microscopy. Immunohistochemical staining detected enhanced endothelial cell expression of intercellular adhesion molecule- 1 (ICAM- 1) and von Willebrand factor (vWF) in the mesenteric arteries and veins. Mast cells were noted to be more prevalent in affected mesenteric tissue from drug-treated animals. The present findings suggest that apoptosis of endothelial and smooth muscle cells, activation of endothelial cells, recruitment of mast cells, and increased expression of adhesion molecules are important factors to the overall pathogenesis of SK&F 95654-induced vasculitis.     

云南省1999-2003年围产儿出生缺陷监测结果分析

目的为了解云南省围产儿出生缺陷的发生种类及分布情况,寻找影响出生缺陷的相关因素.方法 1999年1月-2003年12月监测云南省多所医院住院分娩孕28w-产后7d的围产儿.按全国出生缺陷统一标准要求,由医院逐季上报<围产儿数季报表>,<出生缺陷登记卡>至省妇幼保健院.结果围产儿出生缺陷率为10.26‰(1013/98690),指(趾)畸形;唇腭裂;神经管畸形是云南省围产儿出生缺陷前3位高发种类,男性出生缺陷发生率为10.84‰(561/51732)高于女性9.41‰(442/46958),城市高于乡村,产妇年龄≥35岁是出生缺陷的高发风险因素.结论进一步开展婚前生殖健康教育和医学检查,指导新婚妇女服用小剂量叶酸预防神经管畸形,加强全民健康教育,提高环境意识,做好婚前保健,优生和孕产期保健,开展产前筛查或产前诊断是减少出生缺陷发生的有力措施.