地高辛标记核酸探针技术在家兔视网膜C－sis原癌基因检测中的应用

用ＡＧＰＣ法抽提幼年家兔视网膜总ＲＮＡ，并应用了地高辛标记核酸探针的方法。经斑点杂交显示幼年家兔视网膜出现Ｃ—ｓｉｓ原癌基因表达。对实验方法以及Ｃ－ｓｉｓ原癌基因表达的意义进行了探讨。认为此方法快速、灵敏，检测Ｃ－ｓｉｓ原癌基因表达对深入研究视网膜生长发育及肿瘤等病变有一定意义。     

胎盘型谷胱甘肽S转移酶mRNA在大鼠肝癌癌前病变组织中的原位表达

应用原位杂交技术，观察了二乙基亚硝胺（ＤＥＮ）诱发大鼠肝癌前病变组织中胎盘型谷胱甘肽Ｓ转移酶（ＧＳＴ－Ｐ）ｍＲＮＡ的表达。结果显示，ＧＳＴ－ＰｍＲＮＡ主要在癌前病变肝组织中的变异灶及灶外卵圆形细胞内表达，且在变异灶间或和同一灶内阳性细胞间表达程度不尽一致，而正常肝、再生肝组织中未见其表达。提示在分子水平上变异灶细胞及卵圆型细胞可能成为实验性肝癌的癌前期细胞     

Galanin gene expression declines with adulthood in the cholinergic fields of the horizontal diagonal band of male rats

The neuropeptide galanin (GAL) influences leaming and memory processes, perhaps by inhibiting cholinergic function. We recently reported that, in the rat, the nucleus of the horizontal limb of the diagonal band (HDB) exhibits the highest level of GAL mRNA coexpression by basal forebrain (BF) cholinergic neurons and, in the HDB, virtually all GAL mRNA-expressing neurons correspond to the cholinergic cell type. Since GAL gene expression is induced across puberty in many brain regions, we used in situ hybridization histochemistry and quantitative autoradiography to assess GAL gene expression across the rostro-caudal extent of the HDB in prepubertal and adult male rats and to determine whether GAL gene expression is also regulated during maturation in this BF region. Our results show that the number of GAL mRNA-expressing cells per section is significantly reduced in the HDB with adulthood. Post-hoc analysis indicated that these age-associated differences in the number of GAL mRNA-expressing cells per section could be ascribed to the rostral and central subregions of the HDB. Age-related differences in the labeling intensity of GAL mRNA-expressing neurons were also detected in the rostral and central subregions of the HDB. No age-associated differences in GAL gene expression were found in the caudal HDB subregion. These results suggest that: (1) in contrast to other brain regions, GAL gene expression in the cholinergic BF may be negatively regulated by factors concomitant with puberty; and (2) the inhibition of cholinergic function by cosecreted GAL may be enhanced prior to puberty within cholinergic neurons of the rostral and central aspects of the HDB.     

肝癌组织中HOXA9 mRNA的表达及其临床意义

目的研究肝癌病例癌组织、癌旁组织及非癌肝组织中HOXA9 mRNA的表达及其临床意义.方法标本经10%中性福尔马林固定后常规制作石蜡包埋切片,HOXA9 mRNA行原位杂交检测.结果临床分析发现术后血清AFP值、AFU值、Glo值、PAB值及SHCSP阳性率均有明显减少.癌组织HOXA9mRNA阳性表达评分(1.94±1.69)明显低于非癌肝组织(3.17±1.70),两者有显著差异;癌旁组织HOXA9mRNA阳性表达评分(2.54±1.20)与非癌肝组织及癌组织无显著差异.癌组织、癌旁组织及非癌肝组织三者HOXA9mRNA阳性表达率(54.84%、76.92%、83.33%)无显著差异.除HCC中HOXA9RNA阳性表达评分(1.81±1.78)显著低于MHC(2.75±0.50)外,HOXA9mRNA表达阳性率及阳性评分与癌分化程度、AFP值、SHCSP是否阳性、AFU值、肝硬化有无、转移情况、有无癌栓、大体形态及肿块最大径等临床病理特征均无明显关系.结论本研究结果显示HOXA9mRNA可能与肝癌的发生有密切关系.临床上检测HOXA9mRNA的表达情况对肝癌的早期发现和早期诊断可能有较重要的临床意义,值得深入研究.     

An Integrated Technique for Identification of Differential Genes Expressed in Patients with Cancer

Summary: To develop a method for identification of differential gene expression between different cell populations, several convenient techniques of molecular biology, including subtractive hybridization, suppression PCR, T/A cloning and sequencing, were used to identify genes expressed differentially in CD45^- and CD45^- cells isolated from U266 cell line of multiple myeloma. Our results showed that the levels of abundant genes scale down 20 times through subtractive hybridization.Plasmid DNA from CD45^- cell clones was hybridized with forward or backward cDNA probes synthesized from CD45^- and CD45 cells, respectively. A few of differentially expressed genes reconfirmed by RT-PCR were identified from 500 expressed clones of CD45^- cells. It is concluded that a strategy for gene expression identification developed from conventional molecular biological methods can be used in different laboratories.     

大鼠脊髓损伤修复差异表达cDNA文库的构建

目的　 运用改良消减杂交技术构建大鼠脊髓损伤修复过程中出现的差异表达基因文库 ,为寻找在脊髓损伤后的修复过程中起关键作用的分子提供帮助。方法　在经典消减杂交的基础上引进SMART反转录、长距离PCR、磁性分离系统、离心柱色谱等方法建立改良消减杂交技术 ,通过两轮消减杂交构建差异表达基因的cDNA消减文库 ,并对文库中的差异基因进行批量克隆与生物信息学分析。结果　 将两轮消减杂交后的差异表达基因转化大肠杆菌JM10 9以建立差异表达基因文库 ,库容量大小为 2× 10 3。从文库中随机挑取 90个克隆进行酶切鉴定与测序分析 ,得到 4 0个差异基因序列 ,其中 32个已知序列 ,8个新序列。 结论　 通过改良消减杂交成功建立了脊髓损伤修复过程中出现的差异表达基因文库 ,为筛选在大鼠脊髓损伤修复过程中起关键作用的基因 ,进一步探讨脊髓损伤修复的分子机制奠定了基础     

An alternative mechanism of actions for neuroleptic and antidepressant drugs

It has earlier been proposed by the author that the aetiology of schizophrenic symptomatology may be due to the presence of abnormally connected interhemispheric fibres which link specialised functions in the brains of schizophrenics that are not connected in normal subjects, and that the neuroleptic drugs may produce their action through a local anaesthetic-like effect in suppression of conduction in these fibres. This line of thought has been extended here to consider the possible mechanism of action of the neuroleptic drugs in more detail, as well as that of the tricyclic antidepressant drugs which are derivatives of the phenothiazine group. Pharmacological similarities with the local anaesthetics both structurally and functionally have been considered, as well as the effects that these drug groups may have in common with the lithium salts. It has been suggested that these drugs all produce their primary effect on cell membranes, though not necessarily at the synapse, that the time course of their clinical effect may correlate with their incorporation into various cell membranes within the CNS, and that they may thus bring about a fundamental alteration in cell membrane microstructure. The possible role of electroconvulsive therapy has also been considered. The corollary of this argument is that the affective disorders may be genetically determined diseases of cell membrane microstructure.     

Identification of differentially expressed genes in omental adipose tissues of obese patients by suppression subtractive hybridization

To identify differentially expressed genes between obese individuals and normal control, we have undertaken suppression subtractive hybridization （SSH）. Omental adipose tissues were obtained via abdominal surgery for appendicitis in both 13 obese subjects[BMI （body mass index） 〉 30 kg/m（2）] and 13 normal subjects （BMI 〉 18 and 〈 25 kg/m（2））.