反义核酸对肿瘤细胞多药抗性的抑制：作用靶点及作用方？…

为了用反义寡聚ＤＮＡ调控要霉素肿瘤细胞ＫＢ－Ａ－１的多药抗性,对反义核酸的预作用位点、作用方式和硫代化类似物进行了研究。筛选得到两段对应于翻译起始区（３）和Ｐ－糖蛋白ＡＴＰ结合位点（９）的２０聚反义片段,发现反义核酸的使用次数对结果的检测有很大影响,采用５μｍｏｌ／Ｌ５μｍｏｌ／Ｌ的反义片段９连续处理４ｄ,实现了将近５０％抑制效果,而其正义和错义链在１０μｍｏｌ／Ｌ沈度的同样处理条件下则没有检测到     

伤寒杆菌耐药质粒pRST98与细菌在巨噬细胞内存活关系的研究

目的　研究伤寒杆菌的不相容性C群接合性耐药质粒 pRST98对其宿主菌在巨噬细胞内存活的影响。方法　用标准系列稀释法检测 3株同源染色体伤寒杆菌———含 pRST98的野生型伤寒菌株STpRST98、经SDS人工消除pRST98的突变体菌株ST(R-)及 pRST98重新导入突变体的接合子 pRST98/ST(R-) ,研究它们在BALB/c小鼠腹腔巨噬细胞M J774A .1内的存活情况。结果　巨噬细胞吞噬后 6 0min内大多数待检菌被杀死 ,但 pRST98宿主菌在巨噬细胞内的活菌数明显高于无此质粒的菌株 (P <0 .0 5 )。结论　pRST98能提高其宿主菌在巨噬细胞内的存活能力。     

胎儿脐动脉血流速度指数的临床应用研究

目的：探讨胎儿脐血流S／D值检测的临床意义。方法：采用SRF608型胎儿脐血流检测仪对600例孕妇进行脐血流检查,判定标准为S／D值大于或等于该孕周正常范围的90th%为异常,即S／D≥3．0。结果：妊高征,宫内发育迟缓（IUGR）、脐绕颈、宫内窘迫等病理妊娠中S／D值≥90th%者与正常组比较发生率明显上升,差异非常显著（P＜0．01）。且上述病理妊娠组中S／D比值≥90th%者分娩新生儿发生窒息率明显高于＜90th%者,差异非常显著（P＜0．01）。结论：脐动脉血流波型检测在指导临床处理及评估胎儿预后方面有着重要的作用。     

Copy number of the 16S rRNA gene in Coxiella burnetii

Coxiella burnetii is an obligate intracellular bacterium with a doubling time of 5–7 hours. Chromosomal DNA from C. burnetii was digested with various restriction enzymes previously determined to not cut within the genomic 16S rRNA gene, or with a combination of these noncutting enzymes in conjunction with AflIII, a restriction enzyme that cuts twice within the 16S rRNA gene. Restriction fragments were resolved electrophoretically and probed with a radiolabeled DNA fragment containing the 3 AflIII portion of the C. burnetii 16S rRNA gene. Only a single DNA fragment in these digests hybridized to the probe, indicating that there is a single genomic copy of the 16S rRNA gene in C. burnetii and thus only a single copy of the rRNA operon.     

I269S Mutation in horse liver alcohol dehydrogenase S isoenzyme and its reactivity for steroids and retinoids

Ile-269 in horse liver alcohol dehydrogenase isoenzyme S(HLADH-S) was mutated to serine by phosphorothioate-based site-directed mutagenesis in order to study the role of the residue in coenzyme binding. The specific activity of the mutant(I269S) enzyme to ethanol was increased 49-fold. All turnover numbers of I269S enzyme toward 9 primary alcohols were increased. The mutant enzyme showed 3.6, 4.6, 11.6-fold higher catalytic efficiency for 5β-androstane-3,17-dione, 5β-cholanic acid-3-one and retinal than wild-type, respectively. The reaction mechanism of I269S enzyme was ordered bi bi as wild-type's. These results indicate that the hydrophobic interaction of Ile-269 residue with coenzyme plays an important role in dissociation of coenzyme from enzyme-coenzyme complex, which has been known as the rate limiting step of ADH reaction.     

Different types of structural variation in STRs: HumFES/FPS,HumVWA and HumD21S11

Summary Alleles of the STR systems HumFES/FPS, HumVWA and HumD21S11 were sequenced and analyzed. Sequence data revealed 3 different systems concerning the complexity of their sequence structure. HumFES/FPS belongs to the STR polymorphism with a simple repeat structure. Only 2 subtypes were found with a base substitution in the 5-flanking region and no variation in the repeat region. In the STR system HumVWA the sequence structure of the repeat region is more complex, because 2 tetranucleotide units TCTA and TCTG were present. Additionally allele 14 revealed a completely different sequence structure leading to a different electrophoretic mobility. The repeat region of HumD21S11 is compound in structure. The possibility of variation at 3 positions leads to the occurrence of microheterogeneities in fragments of apparent length. In the upper allele range alleles arise with an additional incomplete TA-repeat.     

12-Lipoxygenases and 12(S)-HETE: role in cancer metastasis

Arachidonic acid metabolites have been implicated in multiple steps of carcinogenesis. Their role in tumor cell metastasis, the ultimate challenge for the treatment of cancer patients, are however not well-documented. Arachidonic acid is primarily metabolized through three pathways, i.e., cyclooxygenase, lipoxygenase, and P450-dependent monooxygenase. In this review we focus our attention on one specific lipoxygenase, i.e., 12-lipoxygenase, and its potential role in modulating the metastatic process. In mammalian cells there exist three types of 12-lipoxygenases which differ in tissue distribution, preferential substrates, and profile of their metabolites. Most of these 12-lipoxygenases have been cloned and sequenced, and the molecular and biochemical determinants responsible for catalysis of specific substrates characterized. Solid tumor cells express 12-lipoxygenase mRNA, possess 12-lipoxygenase protein, and biosynthesize 12(S)-HETE [12(S)-hydroxyeicosatetraenoic acid], as revealed by numerous experimental approaches. The ability of tumor cells to generate 12(S)-HETE is positively correlated to their metastatic potential. A large collection of experimental data suggest that 12(S)-HETE is a crucial intracellular signaling molecule that activates protein kinase C and mediates the biological functions of many growth factors and cytokines such as bFGF, PDGF, EGF, and AMF. 12(S)-HETE plays a pivotal role in multiple steps of the metastatic cascade encompassing tumor cell-vasculature interactions, tumor cell motility, proteolysis, invasion, and angiogenesis. The fact that 12-lipoxygenase is expressed in a wide diversity of tumor cell lines and 12(S)-HETE is a key modulatory molecule in metastasis provides the rationale for targeting these molecules in anti-cancer and anti-metastasis therapeutic protocols.     

Subtype specificity of γ-aminobutyric acid type A receptor antagonism by clozapine

Clozapine, an atypical neuroleptic, functionally antagonizes the -aminobutyric acid-induced chloride uptake via the main central inhibitory receptor, -aminobutyric acid type A (GABA_A) receptor, in brain vesicles. GABA_A antagonism by micromolar concentrations of clozapine is more efficient in rat cerebrocortical and hippocampal membranes than in cerebellar membranes, as evidenced by clozapine reversal GABA-inhibition of [³⁵S]t-butylbicyclophosphorothionate ([³⁵S]TBPS) binding. A typical neuroleptic, haloperidol, failed to antagonize GABA in any of these brain regions, while the specific GABA_A antagonist 2-(3-carboxy-2,3-propyl)-3-amino-6-p-methoxyphenylpyrazinium bromide (SR 95531) was efficient in all three brain regions. Clozapine action on [³⁵S]TBPS binding was unaffected by the benzodiazepine receptor antagonist flumazenil. Clozapine inhibited the binding of [³H]muscimol and [³H]SR 95531 to the GABA recognition site, but this effect only partially correlated with the regional differences in and the potency of clozapine antagonism of GABA-inhibition of [³⁵S]TBPS binding, suggesting that also other than GABA sites may mediate clozapine actions. Autoradiography of [35S]TBPS binding revealed GABA antagonism by clozapine in most brain regions. Main exceptions were cerebellar granule cell and molecular layers, olfactory bulb external plexiform and glomerular layers and primary olfactory cortex, where clozapine antagonized GABA inhibition less than average, and lateral hypothalamic and preoptic areas where its antagonism was greater than average. Recombinant 622 receptors, the predominant 6 subunit-containing receptor subtype in cerebellar granule cells, failed to show GABA antagonism by clozapine up to 100 M. In contrast, recombinant 122 receptors, forming the predominant receptor subtype in the brain, were clozapine sensitive. Recombinant 622 and 632 receptors resulted in clozapine-insensitive receptors, whereas 612 receptors were clozapine sensitive. The efficacy of clozapine to antagonize GABA in 1x2 receptors decreased in the order of 112>122>132. The results indicate that clozapine antagonizes the function of most GABA_A receptor subtypes, and that the interaction is determined by the interaction of the and subunit variants. GABA antagonism is a unique property of clozapine, not shared by haloperidol, which might be involved in the pharmacological mechanism for the increased seizure susceptibility associated with clozapine treatment.     

Pediatric intestinal transplantation

Advancements in donor management, organ preservation and operative techniques, as well as immunosuppressive therapies, have provided children with intestinal failure and its complications a chance not only for enteral autonomy but also long-term survival through intestinal transplantation (ITx). First described in the 1960’s, experience has grown in managing these complex patients both pre- and post-transplant. The goals of this review are to provide a brief history of intestinal transplantation and intestinal rehabilitation in pediatric patients, followed by focused discussions of the indications for ITx, induction and maintenance immunosuppression therapies, common post-operative complications, and outcomes/quality of life post-transplant.     

46例髓母细胞瘤的免疫组化研究

目的：了解髓母细胞瘤发生、分化及其病理生物学行为,方法：４６ 例手术切除髓母细胞瘤组织用免疫组织化学方法对胶质酸性蛋白（ Ｇ Ｆ Ａ Ｐ）、中间丝结蛋白（ｖｉｍ ｅｎｔｉｎ）、神经纤维（ Ｎ Ｆ）、增殖细胞核抗原（ Ｐ Ｃ Ｎ Ａ）、抑癌基因ｐ５３ 及 Ｓ１００ 蛋白进行检测。结果： Ｇ Ｆ Ａ Ｐ、ｖｉｍ ｅｎｔｉｎ、 Ｐ Ｃ Ｎ Ａ、 Ｓ１００ 蛋白阳性率分别为 ７１．１１％ 、４３．４４％ 、９２．００％ 、５２．１７％ ,阳性指数分别为４６．２５‰、１７．００‰、８７５．４０‰、８．４２‰。 Ｎ Ｆ表达极弱,而ｐ５３ 几乎不表达。结论：髓母细胞瘤的发生可能与 ｐ５３ 抑癌基因突变无明显相关;髓母细胞瘤的多数肿瘤细胞向胶质细胞方向分化;髓母细胞生长迅速以及其高度恶性与 Ｐ Ｃ Ｎ Ａ 检测超常表达相一致。