No evidence for an altered mRNA expression or protein level of haematopoietic cell phosphatase in CD34+ bone marrow progenitor cells or mature peripheral blood cells in polycythaemia vera

Abstract: Polycythaemia vera (PV) is a myeloproliferative disorder characterized by haematopoietic progenitor cells being hypersensitive to cytokines such as erythropoietin, interleukin-3, stem cell factor and insulin-like growth factor 1, which results in an increased production of mature blood cells. The pathogenetic cellular mechanism(s) behind this hypersensitivity to cytokines is unknown, but the number of cytokine receptors and the interaction between ligand and receptor are normal in PV. Interest has therefore focused on post-receptor mechanism(s). Haematopoietic cell phosphatase (HCP) is an intracellular tyrosine phosphatase that has been demonstrated to regulate proliferative signals negatively induced by the cytokines mentioned above. Moreover, motheaten mice that genetically lack HCP have an increased amount of erythroid progenitors that are hypersensitive to Epo, and patients with familial polycythaemia have been shown to exhibit a mutation of the Epo receptor gene that includes the docking site for HCP. We therefore studied mRNA expression of HCP in pure populations of CD34⁺ cells, granulocytes, platelets and lymphocytes from patients with PV, chronic myeloid leukaemia (CML) or essential thrombocythemia (ET), as well as healthy controls. Using a polymerase chain reaction analysis employing specific primers for HCP, we failed to detect any abnormalities of HCP expression in PV in any of the cell populations that were examined. Moreover, HCP mRNA expression was similar in ET and CML compared to controls. Finally, Western blot analysis revealed a normal HCP protein content in PV granulocytes and platelets. We therefore conclude that neither an impaired expression of the HCP gene nor a defect in HCP protein synthesis is present in PV, and does not seem to play a role in the aetiology of this disorder.     

Immunohistochemistry of glial reaction after injury in the rat: Double stainings and markers of cell proliferation

The astrocytic reaction in the rat after brain injury has been studied immunohistochemically for intermediate filaments (GFAP and vimentin), also with double staining procedures, and for markers of proliferation (BrdU and PCNA). GFAP-positive reactive astrocytes appeared around the lesion, where they were vimentin-positive and at a distance. BrdU and PCNA showed a high labelling index around the wound at day 2 and scattered positive nuclei were also found at a distance in the ipsilateral side. BrdU-positive astrocytes represented a minor fraction of GFAP- and vimentin-positive astrocytes. The expression of vimentin persisted at least 15 days after the lesion. Our results could suggest that distant reactive astrocytes originate through hypertrophy while those close to lesion arise by hyperplasia from mature or immature glial cells. The hypothesis is formulated that cells of the periventricular matrix contribute to the post-traumatic proliferative activity.     

聚合酶链反应产物特异性的阳性证实

以聚合酶链反应(PCR)法在mRNA水平检测T淋巴细胞受体α链可变区基因表达为例,介绍用~(32)P标记的人工合成寡核苷酸探针对PCR产物特异性作阳性证实的方法。该法以干琼脂糖凝胶作为支持物、相对较为简便和省财。用Ca探针以干凝胶作支持物的杂交结果,证实29个Vα基因的PCR扩增中物均为特异性的,放射自显影的带型与位置和溴乙锭染色所示完全吻合。     

磷脂酶C与生殖

磷脂酶C(PLC)是磷脂酰肌醇信号转导途径中一个关键酶。目前确认的哺乳类PLC共有PLC β、PLC γ、PLC δ、PLC ε和最近新发现的PLC ζ 5种亚型 ,12种同工酶。不同亚型的结构、调控和组织分布各有差异。PLC在调控信号转导通路中的特性使其在顶体反应、卵子激活中发挥了巨大的作用。本文综述了PLC家族 5种亚型的结构、调控机制及PLC在男性生殖领域的相关内容 ,包括PLC引起卵子发生Ca2 + 振荡、激活卵子、促进胚胎发育等 ,并进一步探讨了PLC在临床上可能的应用前景。     

子宫内膜癌组织中p16 mRNA及P16蛋白的表达

①目的　探讨抑癌基因 p16与子宫内膜癌发生、发展的关系。 ②方法　应用逆转录聚合酶链反应(RT PCR)和免疫组织化学技术 ,对 4 2例子宫内膜癌及 15例正常子宫内膜组织中 p16mRNA及P16蛋白进行检测 ,并结合临床病理特征进行分析。③结果　子宫内膜癌组织中p16mRNA及P16蛋白的阳性表达率分别为6 9.0 5 %和 5 9.5 2 % ,显著低于正常组织 (χ2 =6 .0 15、8.6 2 5 ,P <0 .0 5 )。p16mRNA及P16蛋白的表达缺失与肿瘤的组织学类型无关 (χ2 =0 .10 0、0 .2 94 ,P >0 .0 5 ) ,而与组织学分级和临床分期有关 (χ2 =3.95 4、6 .873,P <0 .0 5 )。④结论　p16mRNA和P16蛋白的表达缺失在子宫内膜癌的发生和发展中可能起重要作用     

等位基因特异PCR技术检测结核分支杆菌rpoB基因突变研究

目的 建立并评价等位特异多聚酶链反应检测结核分支杆菌耐利福平基因rpoB突变的方法。方法 设计与rpoB基因突变后的碱基配对的引物用于扩增突变基因，建立AS－PCR检测rpoB基因突变的方法，并对58株结核菌进行了检测。结果 AS－PCR检测结核菌耐利福平相关基因rpoB的敏感性为77.3%，特异性达91.7%。AS－PCR检测rpoB基因，有1628A→T、1657C→T，G和1673C→T突变分别占耐利福平结核菌株的18.2%，22.7%和36.4%。检测rpoB突变与MIC测定RFP耐药率和PCR－SSCP检测的总符合率分别为86.2%和74.1%。检测试验可在3－4h内完成。结论 AS－PCR方法是检测结核菌耐利福平基因突变的敏感和快速方法，可在临床实验室使用并为临床医师提供治疗依据。     

Value of digital volume tomography in patients with conductive hearing loss

Digital volume tomography (DVT) is an extension of panoramic tomography. With this diagnostic technique, characterized by high resolution, a narrow section width (0.125 mm) and three-dimensional display, small pathological processes can be well visualized. Twenty-five patients with the history of a progressive hearing loss were examined with DVT (Accu-I-tomo, Morita, Japan). The results were compared with pre- and intraoperative findings to evaluate the diagnostic value of DVT in cases of erosion of the ossicular chain. With high resolution and artifact-free demonstration of the middle ear and the ossicular chain, it was possible to define its continuity preoperatively by DVT in all 25 cases. An intact ossicular chain was found by DVT in 13 cases and was later confirmed by surgery. The predicted erosion of the ossicles was verified in 12 patients, and a tympanoplasty type III was performed. Digital volume tomography is an excellent technique to examine the middle ear cleft and inner ear, and expands the application of diagnostic possibilities in the lateral skull base. Therefore, improvement in preoperative diagnosis is achieved along with more accurate planning of the surgical procedure. Digital volume tomography delivers a small radiation dose with a high resolution and a low purchase price for the equipment.     

临床输液反应的分析与预防

目的对临床发生的输液反应进行分析,找出发生的原因,探讨预防措施。方法:对30例输液反应资料进行分析。结果:以热原反应最为多见,占76.67%,而过敏反应,占16.67%,局部反应,占6.67%。结论:严格执行消毒制度,遵守无菌操作规程,把好药品和输液器具质量关,合理用药,注意配伍等是减少临床输液反应的关键。     

脊麻药复合微量芬太尼对防治牵拉反应的探讨

目的 探讨脊麻药中复合微量芬太尼对牵拉反应的防治效果。方法 选择美国麻醉医师协会（american society of anesthesiologists，AsA）Ⅰ-Ⅱ级、足月、初产及单胎而需行剖宫产术的病人60例，随机分成2组，每组30例。2组病人的年龄、性剐、体重、局麻药用量、麻醉方法均无显著性差异，均选用腰硬联合麻醉（combined spinal—epidural anesthesia，CSEA）。A组：等比重0．5％罗哌卡因9-11mg＋芬太尼10μg。B组：等比重0．5％罗哌卡因9-11mg。观察术中牵拉反应和生命体征并记录，进行统计学处理。结果 2组间麻醉前、后相对应的MAP（平均动脉压）、HR（心率）、RR（呼吸频率）和SpO2（血氧饱和度）变化均无显著性差异（JP〉0．05），2组间新生儿Apgar评分均无显著性差异（P〉0．05），比较2组术中的牵拉反应，A组明显优于B组，有显著性差异（P〈0．05）结论 脊麻药中加入微量芬太尼对牵拉反应有良好的防治效果。