端粒酶及nm23的表达与胃腺癌预后的关系探讨

目的 :探讨端粒酶和nm2 3癌基因蛋白表达与胃癌根治术后复发及预后的关系。方法 :采用端粒酶原位分子杂交技术及免疫组化S P法分别对 4 5例原发性胃腺癌组织中端粒酶的活性及nm2 3进行检测 ,并结合内镜及随访资料进行分析。结果 :胃腺癌端粒酶阳性表达率及nm2 3低表达率在有淋巴结转移组显著高于无淋巴结转移组 ;在术后 5年内复发者中显著高于无复发者。虽然随着胃腺癌分化程度的降低及浸润深度的增加 ,端粒酶活性及nm2 3低表达呈增强趋势 ,但其差异无统计学意义。结论 :胃腺癌组织中端粒酶的活性表达及nm2 3的低表达与淋巴结转移及术后复发显著相关 ,两者的表达变化对判断胃癌术后复发及预后判断有重要临床意义     

多发性骨髓瘤负性相关基因DAZAP2蛋白高效表达和纯化

目的在前期的研究中，DAZAP2基因在多发性骨髓瘤（Multiple myeloma，MM）患者中表达明显下调，可能为MM的负性基因．须进一步获得DAZAP2蛋白，制备抗体，从而在蛋白水平上研究DAZAP2的表达、结构和功能．方法将原核重组质粒pQE30－DAZAP2转化大肠杆菌，阳性菌株经1 mmol／LIPTG诱导表达目的蛋白，再经过纯化和透析结果IPTG诱导4hr时，得到大量重组目的蛋白，占可溶蛋白的20％左右，经过Ni—NTA层析柱纯化。获得分子量为17kD的DAZAP2蛋白浓度为0．97mg／m1．结论DAZAP2蛋白获得高效表达，并且建立快速简便的纯化方法，目的蛋白可以用于下一步的实验．     

甲状腺肿的专家诊断与咨询系统

本文通过对１７４８３例甲状腺肿大中五种甲状腺病病例的收集、分析、整理、统计与提炼，确定了诊断原则，建立了知识库、病例库和正反双向混合推理机制，完成了用Ｔｕｒｂｏ－Ｐｒｏｌｏｇ语言编制。采用适用于表示医学产生式知识表示方法所开发的专家诊断与咨询系统，为教学、临床工作及家庭保健提供了有价值的软件。经临床病例验证，该软件准确、可靠、有很强的实用价值，对人工智能的研究、临床及教学的现代化，提高人们的保健水     

Embryonic development of four different subsets of cholinergic neurons in rat cervical spinal cord

The developmental stage at which a neuron becomes committed to a neurotransmitter phenotype is an important time in its ontogenetic history. The present study examines when choline acetyltransferase (ChAT) is first detected within each of four different subsets of cholinergic neurons previously identified in the cervical enlargement of the spinal cord: namely, motor neurons, partition cells, central canal cluster cells, and dorsal horn neurons. By examining the temporal sequence of embryonic development of these cholinergic neurons, we can infer the relationships between ChAT expression and other important developmental events. ChAT was first detected reliably on embryonic day 13 (E13) by both biochemical and immunocytochemical methods, and it was localized predominantly within motor neurons. A second group of primitive-appearing ChAT-positive cells was detected adjacent to the ventricular zone on E14. These neurons seemed to disperse laterally into the intermediate zone by E15, and, on the basis of their location, were tentatively identified as partition cells. A third group of primitive ChAT-immunoreactive cells was detected on E16, both within and around the ventral half of the ventricular zone. By E17, some members of this "U"-shaped group appeared to have dispersed dorsally and laterally, probably giving rise to dorsal horn neurons as well as dorsal central canal cluster cells. Other members of this group remained near the ventral ventricular zone, most likely differentiating into ventral central canal cluster cells. Combined findings from the present study and a previous investigation of neurogenesis (Phelps et al.: J. Comp. Neurol. 273:459-472, '88), suggest that premitotic precursor cells have not yet acquired the cholinergic phenotype because ChAT is not detectable until after the onset of neuronal generation for each of the respective subsets of cholinergic neurons. However, ChAT is expressed in primitive bipolar neurons located within or adjacent to the germinal epithelium. Transitional stages of embryonic development suggest that these primitive ChAT-positive cells migrate to different locations within the intermediate zone to differentiate into the various subsets of mature cholinergic neurons. Therefore, it seems likely that spinal cholinergic neurons are committed to the cholinergic phenotype at pre- or early migratory stages of their development. Our results also hint that the subsets of cholinergic cells may follow different migration routes. For example, presumptive partition cells may use radial glial processes for guidance, whereas dorsal horn neurons may migrate along nerve fibers of the commissural pathway. Cell-cell interactions along such diverse migratory pathways could play a role in determining the different morphological, and presumably functional, phenotypes expressed by spinal cholinergic neurons.     

Cholecystokinin and neurotensin mRNAs are differentially expressed in subnuclei of the ventral tegmental area

Immunohistochemical studies of ventral tegmental area (VTA) neurons indicate that individual cells can contain dopamine as well as the neuropeptide neurotransmitters cholecystokinin (CCK) and neurotensin (NT). We have defined the distribution of the cells expressing the mRNAs encoding these two dopamine cotransmitter peptides in each of the subnuclei of the ventral tegmental area, and quantitated the extent of expression of each gene by using in situ hybridization methods. These studies reveal significant differences in the patterns of expression of each of these two genes within various subdivisions of the VTA. The rostral linear nucleus contained numerous CCK positive cells, some of which appeared to express preproCCK-mRNA at a very high level, but this nucleus contained relatively few NT-expressing cells. The parabrachialis pigmentosus contained numerous NT and CCK positive cells. The paranigralis and interfascicularis nuclei displayed positive CCK cells but with expression at only modest levels. NT cells were very few in these nuclei. The caudal linear nuclei contained the highest number of NT-expressing neurons and these cells expressed very high levels of NT mRNA. The selective distribution of these peptide genes within the VTA subnuclei may have specific consequences. Studies of the connectivity of neurons in the VTA show that the different subnuclei of this region project to several functionally and architectonically different regions of the cerebral cortex and subcortically to nuclei related to the limbic system. Results from our study show very prominent expression of CCK mRNAs in those subnuclei that project heavily to the prefrontal, other cortical areas, and the amygdaloid complex. The NT gene is expressed prominently in those subnuclei of VTA that project heavily to the entorhinal cortex and amygdaloid complex. These results provide support for a differential role for the NT-expressing neurons than that of CCK-expressing neurons of VTA in "reward" mechanisms and in drug-seeking and motivational behavior. These observations could be applied to create working hypotheses and experimental paradigms to test the differential functional activity of the subdivisions of VTA and their potential roles in the pathogenesis and treatment of drug-seeking behavior and other neuropsychiatric disorders.     

重组人钙调素对白芷细胞增殖及结核杆菌生长的影响

目的 :探讨重组人钙调素 ( rh Ca M)对白芷悬浮细胞及结核杆菌增殖作用的影响。　方法 :用基因重组技术获得人钙调素基因 ( h Ca M c DNA)重组表达质粒 h Ca M / p BV2 2 0 ,将其转化大肠杆菌 DH5α。用 Phenyl-Sepharose CL -4 B疏水亲和层析法纯化重组菌超声上清表达产物。将纯化 rh Ca M加入悬浮培养的白芷细胞及结核杆菌培养基中 ,观察 rh Ca M对其增殖作用的影响。　结果 :含 h Ca M / p BV2 2 0的重组菌经温度诱导可高效表达Ca M蛋白 ,经 15 % SDS-PAGE分析 ,可观察到一相对分子质量为 170 0 0的表达条带 ,Western blot结果证实 ,此表达条带可与标准鼠抗 Ca M Mc Ab起特异反应。每 1L菌液经 Phenyl-Sepharose CL -4 B疏水亲和层析法纯化可获Ca M纯品 3～ 4mg。rh Ca M对白芷细胞增殖作用影响的研究结果表明 ,rh Ca M在低细胞密度培养条件下对白芷细胞有促进增殖作用 ,效果高于标准植物 Ca M。本研究同时还发现一定浓度的纯化 rh Ca M( 1、10μg/ ml)可促进牛型结核杆菌的生长。　结论 :rh Ca M对植物细胞和细菌均有细胞外促增殖作用     

视网膜母细胞瘤中Fas和bcl-2的表达及意义

目的　探讨Fas及bcl 2在视网膜母细胞瘤的表达及意义。方法　用免疫组织化学SP方法检测了 45例视网膜母细胞瘤及 12例正常视网膜组织中Fas及bcl 2蛋白的表达 ,并进行比较。结果　Fas在视网膜母细胞瘤组织及正常视网膜组织的表达分别为 0 .2 5 5± 0 .441,0 .0 83± 0 .2 89,两者比较差异无显著性 (P =0 .114 8) ,均呈低表达 ;bcl 2在视网膜母细胞瘤中的表达为2 .70 2± 1.0 82 ,与正常视网膜组织中 0 .16 7± 0 .389相比差异具非常显著性 (P <0 .0 0 0 1)。结论　Fas在视网膜母细胞瘤及正常视网膜中均呈低表达 ;而bcl 2在视网膜母细胞瘤中呈明显高表达 ,其可明显抑制凋亡的发生 ,与肿瘤的发展关系密切。     

间歇性气囊挤压大鼠腿部对挤压部位和远端骨骼肌一氧化氮合酶mRNA表达的影响

目的:为了进一步了解间歇性气囊挤压法(Intermittent pneumatic compression, IPC)挤压大鼠腿部与一氧化氮(NO)的关系.方法:检测了大鼠骨骼肌中3种一氧化氮合酶(NOS)同工酶:神经型NOS(nNOS);诱导型NOS(iNOS)和内皮细胞型NOS(eNOS)mRNA在IPC作用后的表达变化.25只SD大鼠被随机分为3个模拟实验组和4个IPC实验组.每只鼠取右侧胫前肌(AT)和提睾肌(CM)作为正常对照.IPC组挤压0.5,1,和5h,及挤压5h加等待4h,模拟实验组除不挤压外,其他操作均与实验组相同,然后分离左侧AT和CM作为处理后样品.所有样品应用RT-PCR进行NOS mRNA测定.以样品中看家基因2,3-二羟基丙醛-3-磷酸脱氢酶(GAPHD)cDNA为内参,与NOS cDNA 共同扩增.PCR产物电泳条带密度用NIH图像分析软件定量,并以与正常对照的对比值作为变化比率.结果:在IPC作用0.5、1和5h后,eNOS mRNA显著上升,在AT中分别达到正常对照的1.2,1.8和2.6倍;在CM中分别达到1.2,1.8和2.7倍,而其他NOS,除5hIPC组的nNOS外,总体表现下调.在IPC作用1h加等待4h组中,eNOS mRNA回复至正常对照水平.结论:该结果证实了IPC产生的机械压力至少部分增加了血管壁的剪切压,使内皮细胞增加了NO产物的释放量,导致了挤压部位及远端肌肉的血管扩张和改善了微循环.     

神经生长因子对儿童癫痫的治疗作用

目的 研究神经生长因子（ＮＧＦ）对儿童癫痫的治疗作用。方法 ＮＧＦ２ｍｌ（１０００ｕ）每日或三日一次肌注,１４天～６个月为一疗程。结果 ８０例中总显效率为５８．８％,其中４５例常规抗痫药无效者加用ＮＣＦ后３９例得到控制,显效率８６．６％,３５例单用ＮＧＦ治疗者有效率为９７．１％。结论 ＮＧＦ对儿童癫痫确有较好的治疗作用。