17-β-雌二醇对大鼠垂本前叶细胞转化生长因子基因表达的影响

实验采用无血清原代培养大鼠垂体前叶细胞及原位杂交方法，观察了不同剂量（１０１０、１０８和１０６ｍｏｌ／Ｌ）的１７β雌二醇（ｅｓｔｒａｄｉｏｌ，Ｅ２）对大鼠垂体前叶细胞转化生长因子α（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒα，ＴＧＦα）和转化生长因子β１（ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒβ１，ＴＧＦβ１）基因表达的影响。结果表明：１０１０ｍｏｌ／ＬＥ２作用２４ｈ后，对两种生长因子的表达均无显著影响；而１０８ｍｏｌ／Ｌ和１０６ｍｏｌ／ＬＥ２则显著升高ＴＧＦαｍＲＮＡ，分别为对照组的１５倍和１６倍（ｐ＜０００１）；同时明显降低ＴＧＦβ１ｍＲＮＡ，使ＴＧＦβ１ｍＲＮＡ水平分别降低为对照组的７０％和５５％（ｐ＜０００１）。说明一定浓度的Ｅ２对正常大鼠垂体前叶细胞ＴＧＦα和ＴＧＦβ１基因表达有明显影响，提示这两种生长因子可能以自分泌／旁分泌机制，参与Ｅ２对垂体前叶细胞功能的部分作用     

大鼠肝纤维化Ito细胞纤维连接蛋白受体α5β1的表达

目的研究纤维连接蛋白受体α５β１在大鼠肝纤维化中的作用。方法应用Ｎｏｒｔｈｅｒｎ印迹杂交及免疫组化方法观察大鼠实验性纤维化肝组织和离体Ｉｔｏ细胞纤维连接蛋白（ＦＮ）及其受体α５β１表达的变化。结果（１）α５β１主要分布于血窦壁的内皮细胞和部分结蛋白（ＤＭ）阳性细胞（Ｉｔｏ细胞）。实验组ＤＭ阳性细胞α５β１表达增强，１０周时达高峰。ＦＮ的变化与之同步。（２）实验组肝组织ＦＮ、α５和β１ｍＲＮＡ含量增高，峰值在第６周；离体Ｉｔｏ细胞三种ｍＲＮＡ含量实验组高于对照组。结论结果提示：Ｉｔｏ细胞表达α５β１，肝纤维化时Ｉｔｏ细胞的激活，可导致ＦＮ、α５和β１ｍＲＮＡ表达增强，应用Ｎｏｒｔｈ－ｅｒｎ印迹杂交技术检测ＦＮ及其受体ｍＲＮＡ的表达，从基因水平了解Ｉｔｏ细胞激活和增生状态，能更敏感地反映肝纤维化的发展倾向。     

大鼠β-防御素rBD-1基因在皮肤和肾脏固有表达

设计大鼠β－防御素ｒＢＤ－１ｃＤＮＡ序列一对特异引物：Ｒ１５′→３′ＡＣＴＣＴＧＧＡＣＣＣＴＧＡＣＴＴＴＣＡＣＣＧ;Ｒ２５′→３′ＣＣＣＴＴＧＣＴＴＧＴＣＣＴＴＴＡＴＧＴＣＣ。根据大鼠β－ａｃｔｉｎｃＤＮＡ序列设计一对阳性对照特异引物：Ｂ１５′→３′ＡＧＣＴＧＡＧＡＧＧＧＡＡＡＴＣＧＴＧＣＧ;Ｂ２５′→３′ＧＴＧＣＣＡＣＣＡＧＡＣＡＧＣＡＣＴＧＴＧ。引物由Ｇｉｂｃｏ公司合成。从Ｗｉｓｔａｒ大鼠皮肤、肾脏、气管、子宫颈、膀胱、小肠、脾脏、骨骼肌、骨髓及腮腺组织中提取总ＲＮＡ,行ＲＴ－ＰＣＲ扩增…     

淫羊藿甙逆转转化生长因子β2对LAK,CD3AK细胞的免疫抑 …

目的 探讨淫羊藿甙逆转转化生长因子β２（Ｔｒａｎｓｆｏｒｍｉｎｇ Ｇｒｏｗｔｈ Ｆａｃｔｏｒ β２ＴＧＦβ２）对ＬＡＫ、ＣＤ３ＡＫ细胞的免疫抑制作用及其机制。方法 采用ＭＴＴ法,ＲＴ－ＰＣＲ和流式细胞术。结果 淫羊藿降转受ＴＧＦβ２抑制的ＬＡＫ和ＣＤ３ＡＫ细胞的杀伤活性,并能部分恢复受ＴＧＦβ２抑制的ＬＡＫ细胞表面ＩＬ－２Ｒα表达和ＣＤ３ＡＫ细胞内穿孔素ｍＲＮＡ水平,以及ＣＤ３ＡＫ细胞的增殖活性。     

转化生长因子-α、表皮生长因子受体与胃癌发生的关系及其与增殖细胞核抗原的相关性分析

目的研究在胃癌发生的不同阶段转化生长因子－α（ＴＧＦ－α）、表皮生长因子受体（ＥＧＦＲ）的表达情况及与增殖细胞核抗原（ＰＣＮＡ）表达的关系。方法应用免疫组化ＬＳＡＢ法。结果（１）ＴＧＦ－α在癌周正常粘膜、肠化生组织中的表达明显高于非癌正常粘膜及肠化生（Ｐ＜０．０１）。（２）ＥＧＦＲ在肠化生、不典型增生粘膜表达较正常、癌组织明显升高（Ｐ＜０．０１）。（３）ＴＧＦ－α、ＥＧＦＲ共同表达常伴不典型增生。（４）ＴＧＦ－α、ＥＧＦＲ表达与ＰＣＮＡ表达有明显的相关性。（５）ＴＧＦ－α、ＥＧＦＲ、ＰＣＮＡ表达与肿瘤外侵、淋巴结转移无关。结论ＥＧＦＲ／ＴＧＦ－α是胃癌前病变的一项有意义的标志，结合ＰＣ－ＮＡ监测高危人群可能有助于发现早期胃癌。     

肺巨细胞癌不同转移亚克隆生长因子表达及反应性差异的探讨

目的研究肺巨细胞癌高转移亚系ＰＧｂＥ１和中度转移亚系ＰＧＬＨ７细胞之间部分生长因子的表达及反应性差异。方法利用ＲＴ－ＰＣＲ技术检测了生长因子ＴＧＦα、ＴＧＦβ１、ＩＬ－６、ＩＬ－８、ｂＦＧＦ和ＡＮＧ及受体ＥＧＦＲ、ＩＬ－６Ｒ和ＩＬ－８Ｒ的表达状况;其次采用３Ｈ－ＴｄＲ掺入法观察了重组ＴＧＦα、ＴＧＦβ１和ＩＬ－６对此两个细胞生长的影响。结果ＰＧｂＥ１细胞中ＴＧＦα、ＥＧＦＲ、ＩＬ－６和ＩＬ－６Ｒ的表达水平明显高于ＰＧＨ７细胞,而ＴＧＦβ１、ｂＦＧＦ、ＩＬ－８、ＩＬ－８Ｒ和ＡＮＧ的表达在两个细胞间无明显差别;重组ＴＧＦα和ＩＬ－６对两个细胞均具有生长刺激作用;ＴＧＦβ１具有双重效应。结论ＴＧＦα、ＴＧＦβ１、ｂＦＧＦ、ＩＬ－６、ＩＬ－８、和ＡＮＧ等生长因子可能以自分泌方式参与肺巨细胞癌的生长增殖调控,而ＴＧＦα和ＩＬ－６在肺巨细胞癌的转移中发挥更为重要的作用。     

血管紧张素原基因M235T分子变异与2型糖尿病肾病的关系

目的　探讨血管紧张素原（ａｎｇｉｏｔｅｎｓｉｎｏｇｅｎ , Ａ Ｇ Ｔ） 基因 Ｍ２３５ Ｔ 分子变异与中国人无肾病并发症的２ 型糖尿病（ｄｉａｂｅｔｅｓ ｍ ｅｌｌｉｔｕｓ , Ｄ Ｍ） 、２ 型糖尿病肾病（ｄｉａｂｅｔｉｃ ｎｅｐｈｒｏｐａｔｈｙ , Ｄ Ｎ） 的关系。方法　用 Ｐ Ｃ Ｒ及 Ｒ Ｆ Ｌ Ｐ 方法对８４ 例 Ｄ Ｍ、９６ 例 Ｄ Ｎ 及９８ 名正常对照进行了 Ａ Ｇ Ｔ 基因 Ｍ２３５ Ｔ 多态性的检测。结果　 Ｄ Ｎ 组 Ｔ 等位基因频率０８２ , Ｔ Ｔ 基因型频率０７０ ,与对照组（０６３ ,０４３） 比较有显著差异（ Ｐ＝ ０００３ , Ｐ＝００００４） ;校正了 Ｄ Ｎ 的几种危险因素后, Ｔ Ｔ 基因型对 Ｄ Ｎ 的 Ｏ Ｒ 为３４７（９５ ％ Ｃ Ｉ 为１５１ ～７９４ , Ｐ＝０００３３） 。 Ｄ Ｍ 组基因型频率分布与对照组比较无显著差异（ Ｐ＞ ００５） 。结论　 Ａ Ｇ Ｔ 基因 Ｔ Ｔ 型可能是中国人群２ 型糖尿病肾病的独立危险因素之一。     

GST-人可溶性成纤维细胞生长因子受体1融合蛋白在酵母细胞中的表达及活性测定

目的：表达重组人可溶性成纤维细胞生长因子受体１（ｓｏｌｕｂｌｅ　ｆｉｂｒｏｂｌａｓｔ　ｇｒｏｗｔｈ　ｆａｃｔｏｒ　ｒｅｃｅｐｔｏｒ　１，ｓＦＧＦＲ１），研究其对成纤维细胞生长因子（ＦＧＦ）生物学活性的拮抗作用。方法：采用逆转录－ＰＣＲ（ＰＴ－ＰＣＲ）技术自人肺成纤维细胞获得ｓＦＧＦＲ１　ｃＤＮＡ，测序确证后，将其克隆人酵母细胞表达载体ｐＹＥＸ４Ｔ－１；重组质粒转化入酵母细胞（ＤＹ１５０）中进行诱导表达，表达产物经ＳＤＳ－ＰＡＧＥ及 Ｗｅｓｔｅｒｎ　ｂｌｏｔ鉴定。利用 ＮＩＨ３Ｔ３细胞增殖抑制实验检测重组人 ｓＦＧＦＲ１的生物学活性。结果：经 ＣｕＳＯ４诱导，酵母细胞表达出重组谷胱苷肽转移酶（ＧＳＴ）－ｓＦＧＦＲ１融合蛋白，此蛋白在凝胶上表现为 １条约 ６０ｋＤ的阳性区带，在 Ｗｅｓｔｅｒｎ ｂｌｏｔ实验中可被ＧＳＴ特异性抗体识别。重组ＧＳＴ－ｓＦＧＦＲ１融合蛋白的粗提物在体外能够桔抗ＦＧＦ介导的促ＮＩＨ３Ｔ３细胞增殖的活性。结论：重组ＧＳＴ－ｓＦＧＦＲ１融合蛋白在酵母表达系统中得到有效表达，并具有很好的生物活性。     

一种改良的半定量RT PCR方法

选用细胞因子ＩＬ ４ 为靶基因， 以β 肌动蛋白（β ａｃｔｉｎ） 基因作为内参照， 将两者在同一ＰＣＲ体系中扩增， 分析ＰＣＲ扩增的动力学， 内参照β ａｃｔｉｎ 的引物于ＰＣＲ扩增中的滞后加入， 弥补了ＩＬ ４ 与ＲＴ ＰＣＲ模板量的差异， 使二者的扩增同时处于指数扩增期， 其扩增产物均可在溴乙锭染色的聚丙烯酰胺胶上检测出， 从而可求出ＩＬ ４ 的ｃＤＮＡ 拷贝数占β ａｃｔｉｎ拷贝数的百分比。重复实验的ＣＶ 值在１０％ 以内， 表明此半定量ＲＴ ＰＣＲ 方法的可靠性。     

以Bacmid-杆状病毒-昆虫细胞系统表达人FGF-9

目的：在Ｂａｃｍｉｄ－杆状病毒－昆虫细胞系统中表达人ＦＧＦ－９。方法：采用ＲＴ－ＰＣＲ技术,自新鲜人脑胶质瘤组织获取人ＦＧＦ－９全编码区ｃＤＮＡ,将其克隆入ｐＣＲ＾ＴＭⅡ质粒及ｐＹＥＸ４Ｔ－１真核表达质粒,经ＤＮＡ自动测序仪进行ＤＮＡ序列测定。将人ＦＧＦ－９ｃＤＮＡ定向克隆入ｐＦａｓｔＢａｃ质粒,进一步将其转座入Ｂａｃｍｉｄ中,在昆虫细胞Ｓｆ９中进行表达,采用ＳＤＳ－ＰＡＧＥ对表达产物进行分析。     

Cytological, histological, and immunohistochemical findings of pulmonary carcinomas with basaloid features

Pulmonary basaloid carcinoma (BC), a variant of large cell, nonsmall cell carcinoma (NSCC), and basaloid squamous cell carcinoma (BSQCC) can show features similar to small cell carcinoma (SCC) and large cell neuroendocrine carcinoma (LCNEC). Distinction from SCC, especially on FNA, is therapeutically relevant. We describe cytological, histological, and immunohistochemical features of BC and BSQCC. Numerous cytologic features were documented in cytologic preparations. Similar features and architecture were evaluated in the resections. Immunohistochemical results were recorded. Histologically confirmed BC (n = 3) and BSQCC (n = 3) were included. Five FNAs of SCC, (four with histologic follow-up) were studied for comparison of cytological, histological, and immunohistochemical findings. In cytologic preparations of BC/BSQCC, cells were arranged mostly as tightly cohesive clusters (n = 4) or singly and in clusters (n = 2) with a predominance of clusters. Cytologic features of BC and BSQCC were similar: palisading (n = 6), crush artifact (n = 6), hyperchromasia (n = 5), focal nuclear molding (n = 6; very rare in 2/6), nucleoli, usually pinpoint (n = 3), scant cytoplasm (n = 6), necrosis (n = 5), apoptosis (n = 4), squamous differentiation (n = 1). BSQCC tended to have occasional larger cells, including keratinizing cells in one case. Histologic sections (n = 6) showed neuroendocrine features, including organoid arrangements, nests, and palisading. BC and BSQCC show overlapping features with SCC and LCNEC in cytological and histological specimens. Unlike SCCs, BC/BSQCC lack prominent nuclear molding, show tightly cohesive cell clusters, and demonstrate palisading. However, immunostains were the very helpful and probably necessary to accurately diagnosing BC/BSQCC, which show the immunostaining pattern of p63 (+), HMWCK (+), and TTF-1 (-).     

大鼠旁巨细胞外侧核呼吸神经元自发放电的类型

在32只麻醉、自主呼吸的SD大鼠,用细胞外记录方法,在旁巨细胞外侧核尾侧半(cPGCL)记录到181个自发单位放电。结果发现cPGCL区存在着非呼吸相关神经元(NRRNs,134/181个,占被测试神经元的74％)和呼吸相关神经元(RRNs,47/181个,占26％)。RRNs可分为递增型吸气神经元(I—Aug,35/47个,占74.7％)、递减型吸气神经元(I—Dec,2/47个,4.2％)、稳定型或钟型吸气神经元(I—Con or I—bell,2/47个,4.2％)、递减型呼气神经元(E—Dee,4/47个,8.5％)、稳定型或钟型呼气神经元(E—Con or E—bell,2/47个,4.2％)、稳定型呼—吸跨时相神经元(EINs,2/47个,4.2％)六种亚型。RRNs以递增型吸气神经元为主,主要分布在cPGCL背外侧。(PGCL区RRNs在存在提示该区参与呼吸的调控。)     

Antineutrophil cytoplasmic autoantibody in the absence of Wegener's granulomatosis or microscopic polyangiitis: implications for the surgical pathologist.

Antineutrophil cytoplasmic antibodies (ANCA) are useful serologic markers for the diagnosis and management of patients with Wegener's granulomatosis (WG) and microscopic polyangiitis (MPA). However, problems in diagnosis and classification may occur when patients with other disorders develop ANCA. A 7-year review (1993-1999) disclosed 247 patients whose sera tested positively for ANCA by an indirect immunofluorescence method: 166 patients for cytoplasmic-ANCA (C-ANCA) and 81 patients for perinuclear-ANCA (P-ANCA) Twenty-seven patients had active pulmonary disease and underwent open-lung biopsy or transbronchial biopsy. Eight patients (30%) had a disease other than WG or MPA, and their clinical, pathological, and serological findings were reviewed. The patients, all women, ranged in age from 28 to 77 years (median, 37 y). Dyspnea (n = 6), cough (n = 6), chest pain (n = 2), and/or hemoptysis (n = 2) were present. The duration of symptoms lasted from 3 weeks to 6 years (median, 6 mo). ANCA titers were C-ANCA (n = 4; range, 1:40-1280) or P-ANCA (n = 4; range, 1:40-640). The lung biopsies disclosed nonspecific interstitial pneumonia (n = 4), bronchiolitis obliterans organizing pneumonia (n = 1), diffuse alveolar damage (n = 1), organizing diffuse alveolar hemorrhage without capillaritis (n = 1), and necrotic granuloma (n = 1). No cases showed characteristic histology for WG or MPA. The final diagnoses were various connective tissue disorders (n = 5), chronic hypersensitivity pneumonia (n = 1), postinfectious bronchitis/bronchiectasis (n = 1), and ulcerative colitis-related lung disease (n = 1). Surgical pathologists should be aware that significantly elevated ANCA titers may be associated with diverse forms of pulmonary disease. ANCA positivity alone, in the absence of appropriate clinical or pathologic findings, should not be used to substantiate a diagnosis of WG or MPA.     

35例精神分裂症患者神经心理学评估的对照研究

目的：用神经心理测验评估35例精神分裂症患者认知功能损害的特点。方法：采用多种神经心理测验评估35例精神分裂症患者（12例未用药，8例用药1周以内，15例用药1月以上）和20例健康对照组的学习、记忆、工作记忆、注意、信息处理速度、词语流畅、执行功能等认知功能。对未用药和用药1周以内的精神分裂症患者进行随访。结果：在成套神经心理测验的各个分测验中，精神分裂症患者（未用药、用药1周以内和用药1月以上）成绩均较正常人差[如韦氏数字符号得分未用药者（63．0±13．8），正常对照组（87．10±13．16），P〈0．001]。用药1月以上组的空间广度测验成绩优于未用药组[逆行得分成绩用药1月以上组（7．47±1．81），未用药组（5．50±1．73），P〈0．01；总分成绩用药1月以上组（15．33±3．31），未用药组（12．42±3．65）]。随访研究中，霍普金斯词汇学习测验一修订版、简易视觉空间记忆测验-修订版和视觉空间广度测验的随访成绩优于初测成绩[如空间广度测验的逆行得分初测成绩（5．7±1．5），随访成绩（6．8±1．0，P〈0．05）]。结论：精神分裂症患者存在大脑广泛的认知功能损害。非经典抗精神病药物治疗可能对精神分裂症患者的言语记忆、视觉空间记忆和视觉工作记忆有所改善。     

Heart tumors in children and adults: clinicopathological study of 59 patients from a surgical center.

BACKGROUND: Heart tumors are rare lesions with variegated histological types. Their clinicopathological features could be more comprehensively categorized. METHODS: This is a 19-year retrospective study of 17 infants/toddlers (<2 years of age) and 42 patients aged between 14 and 79 years (mean = 51.5) in a surgical center. RESULTS: Congenital tumors (n = 17; 29%), including rhabdomyomas (n = 9), ventricular fibromas (n = 6), and hemangiomas (n = 1), required surgery mainly because of mass effect. Familial myofibromatosis was the only embolic congenital lesion. Acquired benign tumors (n = 28; 47%) included myxomas (n = 21), fibroelastomas (n = 3), myofibroblastic inflammatory tumors (n = 2), and lipomas (n = 2). Eight (29%) were revealed by systemic embolization. These benign noncongenital tumors were all treated by complete resection, except for an incompletely resected lipoma of the mitral valve. Postoperative arrhythmia (n = 1) and pericardial effusion (n = 3) were the only complications. Primary sarcomas (n = 8; 14%) were mostly vascular tumors (five of eight), and patients with high-grade tumors had a mean survival of 15 months (n = 5). Cardiac metastases (n = 6; 10%) were from carcinomas (n = 3) or sarcomas (n = 3); apart from a necrotic metastasis, all patients died (mean survival of 6 months). CONCLUSIONS: This study shows that, regardless of patients' age, heart tumors can be classified as: (a) congenital lesions, which are spontaneously nonprogressive or regressive lesions possibly requiring surgery mainly because of mass effect; (b) acquired benign tumors, which are lesions requiring surgery often because of embolization risk; and (c) primary and secondary malignant tumors, which are lesions with globally poor prognosis but with some indications for resection.     

The ex-vivo-shunt-model: novel approach for assessing the thrombogenicity of vascular implants

OBJECTIVE: Disadvantages associated with commercially available vascular implants necessitate alternative strategies to develop new vascular prostheses. Although many tissue characterizing strategies have been defined, no valid test for thrombogenicity exists. Here we introduce a novel concept for thrombogenicity testing of vascular implants METHODS: Silastic tubes were implanted into the carotid arteries of 12 sheep. After placing these shunts, tc99m-labeled platelets were administered and test-vessels were put in between the shunts. Native autologous (n=6), as well as native/acellularized allogeneic (n=6/n=6), and xenogeneic (n=6/n=6) carotid arteries and allogeneic (n=6/n=6) and xenogeneic (n=6/n=6) carotid arteries reseeded with allogeneic endothelial-cells, fibroblasts and myocytes were evaluated. Number and time course of intra-operatively deposited platelets were evaluated with a Geiger-counter; certain areas of platelet deposition located, envisioned and characterized by a gamma-camera and scanning electron-microscopy afterwards. RESULTS: Counter results revealed no significant different platelet depositions when comparing silastic tubes with either autologous or allogeneic native carotid arteries. However, starting 5 minutes after placement, acellularized/reseeded allogeneic (p=0.001/p=0.00004), and xenogeneic (p=0.0001/p=0.01) carotid arteries showed significantly more platelet depositions than native autologous carotides. Moreover, it was possible to show that almost no platelets adhere to native vessels or silastic tubes, thus proving the test method itself. CONCLUSION: The Ex-Vivo-Shunt-Model is a valid method to measure and envision the intrinsic thrombogenicity of vascular implants.     

Early human herpesvirus type 6 reactivation after allogeneic stem cell transplantation: a large-scale clinical study

This study investigated the impact of human herpesvirus type 6 (HHV6) reactivation within 100 days of allogeneic stem cell transplantation (allo-SCT) on patient outcomes. HHV6 plasma loads were monitored weekly by quantitative PCR. Of 235 consecutive patients, 112 (48%) had an early positive HHV6 PCR test (group A) and 123 (52%) did not (group B). HHV6 reactivation was less frequent in patients who received reduced-intensity conditioning (P = .028). In group A, only 6 patients (5%) were asymptomatic; the most common clinical manifestations were fever (n = 60), skin rash (n = 57), diarrhea (n = 51), pulmonary complications (n = 19), and neurologic disorders (n = 12). Compared with the patients in group B, those in group A experienced delayed platelet engraftment (P = .003) and more frequent grade II-IV acute graft-versus-host disease (GVHD) (47% versus 30% in group B; P = .009). In multivariate analysis, the most important factors influencing the development of grade II-IV acute GVHD development were early HHV6 reactivation (P = .03) and unrelated donor status (P < .001). HHV6 reactivation adversely influenced 6-month survival (P = .04). Of?the 38 evaluable patients receiving antiviral treatment, 34 had a significantly decreased HHV6 load. Our findings indicate that HHV6 reactivation after allo-SCT is associated with delayed platelet engraftment, early posttransplantation mortality, and the development of acute GVHD. Careful monitoring of HHV6 by PCR is warranted during the early posttransplantation period.     

ATP and ADP in human pre-embryos

Spare human oocytes and pre-embryos from an in-vitro fertilization (IVF) programme were individually analysed for ATP and ADP content using a bioluminescence method employing the firefly luciferin-luciferase reaction. The ATP content of oocytes that failed to fertilize in vitro was 1.71 +/- 0.28 pmol (n = 10), pronuclear stage ova 1.93 +/- 0.08 (n = 6), 2-cell stage 1.78 +/- 0.20 (n = 7), 4-cell stage 1.73 +/- 0.15 (n = 6), 6-8 cell stage 2.76 +/- 0.53 (n = 8), morula stage 2.36 +/- 0.68 (n = 8), early blastocyst stage 2.08 +/- 0.25 (n = 7) and expanded blastocyst stage 2.26 +/- 0.15 (n = 3) pmol. The ADP content of these pre-embryos was low in all stages with a small increment in the 2-4 cell stages and the blastocyst stage. This gave an elevated ATP/ADP ratio (range 19-92) indicating a good energy status. The sensitive luciferin-luciferase assay may be a tool for studying the energy status of spare oocytes and pre-embryos under different incubation conditions in human IVF programmes.     

A retrospective study of 586 tumours in Brazilian cattle

Records from 6,706 necropsy examinations of cattle performed over a 45-year period were surveyed and 586 cases of neoplasia were identified. The organ system most frequently affected by neoplastic disease (n=139 cases) was the alimentary tract. This finding was attributed to a high incidence of squamous cell carcinoma of the upper alimentary tract associated with the chronic ingestion of bracken fern (Pteridium aquilinum). This carcinogenic plant was also associated with a relatively high incidence (n=35 cases) of urinary bladder tumours (enzootic haematuria). Tumours of the alimentary tract were followed, in decreasing order of frequency, by tumours of the skin and subcutis (n=129 cases), haemopoietic tissue (n=101 cases), the eye and periorbital tissues (n=88 cases), the urinary system (n=44 cases), the female reproductive system (n=21 cases), the endocrine system (n=16 cases), the liver and pancreas (n=12 cases), the nervous system (n=6 cases), the respiratory system (n=6 cases) and the mammary gland (n=1 case). The primary anatomical location of 16 tumours was undetermined.     

The characterization of VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses from field samples using RT-PCR and RFLP analysis

Summary Characterization of the VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses (BRV) from field samples was performed using RT-PCR and RFLP analysis. After RT-PCR amplification of the full length VP7 genes and partial length VP4 genes (nucleotides 1 to 1096), four enzymes,EcoRV,NlaIV,BamHI andHpaII were used for digestion analysis. For VP7, four RFLP profiles were observed after analysis of the digests: they were designated as G6, G6s (subtype, showed about 86% nucleotide and 90% amino acid identity to reference G6 strains), G8 and G10. For VP4, three RFLP profiles were observed: designated as P[1], P[5] and P[11].The G typing analysis of 86 BRV fecal samples from 5 states, representing at least 11 different herds revealed that 60.5% (52/86) were G6, which included G6s (9/52); 19.8% (17/86) were G10; 7% (6/86) were G8; 10.4% (9/86) were G6 and G10 mixtures including two G6s samples; and 2.3% (2/86) were G6 and G6s mixtures. The P typing analysis of the same 86 fecal samples revealed that 64% (55/86) were P[5]; 28% (24/86) were P[11]; 1.2% (1/86) were P[1] and 6 samples (7%) were mixtures of either P[11] or P[5]. When the same samples were analyzed according to G and P type specificity, all possible combinations of G and P types existed in the field. The G6P[5] type was most prevalent and accounted for 46.7% (41/86) of the samples; 12.8% (11/86) were G10P[11]; 7% (6/86) were G10P[5] and an equal number were G6sP[11]. The G6P[11] (n=2), G8P[1] (n=1), G8P[5] (n=1) and G8P[11] (n=3) combinations were also observed. The following mixed BRV infections were observed in the field samples; G6sP[5+11] (n=1), G8P[5+11] (n=1), G6+G10P[5] (n=1) G6+G10P[5+11] (n=2), G6+G6sP[11] (n=1), G6+G6sP[1+11] (n=1), G6s+G10P[11] (n=1) and G6s+G10P[5+11] (n=1). Information on the G and P types and G/P combinations in the field samples should be useful for understanding the epidemiology of BRV and designing vaccination strategies to control BRV in the field.