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.     

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。     

螺旋CT血管造影对肺动脉栓塞的诊断价值

目的 评价螺旋CT血管造影（SCTA）对肺动脉栓塞（PE）的诊断价值。方法 对临床疑诊PE的20例患者进行螺旋CT血管造影成像（SCTA）．结果 20例患者中确诊PE7例。7例PESCTA直接征象：肺动脉主干或分支内混合性、附壁性、中心性充盈缺损。间接征象：局限性肺纹理稀疏，肺动脉高压，右心室增大，胸腔积液等。结论 螺旋CT血管造影具有准确、快速、无创性的优点，对肺动脉栓塞的诊断具有广泛应用价值。     

表达载体介导的反义RNA抑制人巨噬细胞移动抑制因子的表达

目的：研究表达载体介导的反义RNA对人巨噬细胞移动抑制因子(MIF)表达的抑制作用。 方法:用亚克隆技术构建可转录MIF反义RNA的真核表达载体pcDNA3-antiMIF。用lipofectamine2000分别将pcDNA3、pcDNA3-antiMIF转染可表达MIF的HEK293(293-MIF)细胞，用Real-time定量PCR鉴定MIF mRNA表达水平。将pcDNA3-antiMIF转化人脐静脉血管内皮细胞(HUVECs)，建立可表达MIF反义RNA的HUVECs(HUVECs-antiMIF)细胞。将MIF的真核表达载体pSecTag-MIF转染HUVECs-antiMIF，用Real-time定量PCR鉴定MIF mRNA的表达水平。 结果:正确构建了MIF反义RNA的表达载体pcDNA3-antiMIF。MIF 反义RNA对293-MIF细胞中MIF表达的抑制水平达32%（P<0.05）。建立稳定表达MIF反义RNA的HUVECs-antiMIF细胞株。HUVECs-antiMIF中MIF的表达受到抑制，表达水平降低40%（P<0.05）。 结论:表达载体介导的反义RNA能有效地抑制MIF的表达，建立了稳定表达MIF反义RNA的HUVECs。     

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.     

Adult-onset type II citrullinemia and idiopathic neonatal hepatitis caused by citrin deficiency: involvement of the aspartate glutamate carrier for urea synthesis and maintenance of the urea cycle

Citrin is a mitochondrial aspartate glutamate carrier primarily expressed in the liver, heart, and kidney. We found that adult-onset type II citrullinemia is caused by mutations in the SLC25A13 gene that encodes for citrin. In this report, we describe the frequency of SLC25A13 mutations, the roles of citrin as a member of the urea cycle and as a member of the malate-aspartate shuttle, the relationship between its functions and symptoms of citrin deficiency, and therapeutic issues.     

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.     

转染survivin反义mRNA对Jurkat淋巴瘤细胞生长和化疗敏感性的影响

目的 研究转染survivin反义mRNA对Jurkat淋巴瘤细胞生长的影响以及转染后淋巴瘤细胞对化疗药物的敏感性。方法 构建survivin反义mRNA真核表达质粒pcDNA3．1-反义（As）survivin;利用脂质体转染法将其转入高表达survivin mRNA T淋巴母细胞淋巴瘤Jurkat细胞系,用逆转录聚合酶链反应（RT—PCR）、免疫组织化学SP法、Western印迹法检测细胞中survivin表达;用细胞计数、流式细胞术（FCM）检测其细胞生长曲线、细胞凋亡指数,并进行光镜、电镜形态学观察;并对转染pcDNA3．1-Assurvivin前后Jurkat细胞分别加入4-羟基-环磷酰胺（CTX）、甲氨蝶呤（MTX）72h后,常规MTT检测细胞存活率。结果 RT—PCR检测转染pcDNA3．1-Assurvivin后48h、5和6周Jurkat细胞survivin mRNA表达,发现survivin mRNA表达皆低于对照组;转染后survivin蛋白表达也明显降低。转染pcDNA3．1-Assurvivin后Jurkat细胞生长倍增时间（52h）明显延长;用FCM检测细胞凋亡发现,转染pcDNA3．1-Assurvivin后Jurkat细胞凋亡指数[20．2％（48h）]明显高于对照组（转染空质粒和未转染组,2．1％和1．3％）;5和6周为6．2％和6．8％,明显高于未转染细胞（1．3％和1．0％）。光镜、电镜观察见转染细胞出现较多凋亡细胞及一些变性肿胀细胞;MTT检测结果显示Jurkat细胞转染前后,经化疗药物4-羟基-环磷酰胺和甲氨蝶呤作用,转染细胞的抑制率明显大于未转染组,差异有统计学意义（P〈0．05）。结论 survivin基因对Jurkat细胞系的生长起着重要的作用,抑制survivin基因表达在T淋巴母细胞淋巴瘤治疗中可能有重要的意义,该基因似可能作为治疗的靶点。