人巨细胞病毒UL140基因在临床低传代分离株中的多态性研究

目的 研究人巨细胞病毒HCMV-UL140基因在临床低传代分离株中的多态性及其与：病性的关系。方法 对40株HCMV临床低传代分离株进行UL140基因全序列的PCR扩增，对121进行了测序及结果分析。结果 40株HCMV-UL140基因PCR扩增均阳性。测序的12株HCMV-UL140开放阅读框架(ORF)均在Toledo株ULl40-ORF的第174位核苷酸处，插入一个胞嘧啶核苷酸．造成移码突变。与Toledo株相比，临床分离株新增了SCAMP磷酸化SPS和酪蛋白激酶Ⅱ磷酸化KP两种重要功能位点，且其所位于的氨基酸位点可能是UL140蛋白的蛋白质作用位点。结论 临床分离株UL140基因的ORF较Toledo株多出231个核苷酸，故两者的核苷酸及其编码产物氨基酸序列明显不同。临床分离株存在SPS及CKP两种新的重要功能位点，可能在HCMV-UL140编码蛋白的生物学功能方面起重要作用。     

人巨细胞病毒UL142基因在临床低传代分离株中的多态性

目的研究人巨细胞病毒(HCMV)UL142基因在低传代分离株中的多态性,探讨其多态性与HCMV先天性感染不同致病性之间的关系。方法27株经荧光定量PCR方法检测HCMVDNA阳性的低传代分离株进行UL142全序列PCR扩增,阳性13株PCR产物进行UL142基因测序及结果分析。结果27株分离株UL142PCR扩增,13株阳性,阳性率48.2%,以Toledo株为参考株序列比较表明,13株UL142可读框长度均与Toledo株相同,为921bp,编码307个氨基酸的蛋白。DNA序列变异均为核苷酸替换,不同临床分离株UL142基因与Toledo株进行同源性比较,结果在核苷酸水平为94.4%~95.9%,氨基酸水平为89.9%~99.6%。二级结构预测分为三种构象。大多数HCMVUL142蛋白重要功能基团位点在所有分离株中均高度保守,仅四个位点在一些分离株中存在缺失或新增。系统进化树分析除Toledo株外,13株核苷酸及氨基酸序列可分为3个基因组。结论13株临床低传代分离株HCMVUL142基因DNA及其编码产物的氨基酸序列比较保守,但仍存在一定多态性。未发现不同临床分离株UL142基因多态性与HCMV先天性感染不同疾病表现的关系。     

人巨细胞病毒UL134基因在临床低传代分离株的多态性研究

目的：人巨细胞病毒（HCMV）呈现一定的基因多态性。病毒的致病性可能与不同分离株的基因变化有关。HCMV Toledo株和其他低传代临床分离株基因组UL/b′区中发现的19个基因（UL133-UL151）可能在病毒潜伏、复制、逃避机体免疫和不同组织细胞嗜性等方面起重要作用。该文研究HCMV不同临床分离株UL134基因及其编码蛋白氨基酸序列的多态性，分析这种多态性与人巨细胞病毒感染引起的各种疾病之间的关系,提供了HCMV基因多态性的一项基本数据。方法：对荧光定量PCR方法检测HCMV-DNA阳性的32株临床低传代分离株进行UL134基因全序列PCR扩增、测序及分析。结果：32株临床低传代分离株UL134基因PCR扩增均阳性。与Toledo株比较，临床分离株UL134基因核苷酸和预测编码蛋白质的氨基酸序列较保守，同源性分别为96.4%~98.3%和92.7%~94.9%，但预测编码蛋白质新增一个SUL位点。结论： 与Toledo株比较，临床分离株UL134基因比较保守，但仍具有一定的多态性, 未发现UL134基因与HCMV致病性之间的本质联系。［中国当代儿科杂志，2007，9（6）：583-586］     

引起不同临床表现的巨细胞病毒UL144基因多态性的研究

目的研究不同临床背景的人巨细胞病毒（HCMV）临床株中UL144基因的多态性，探讨其多态性与HCMV感染临床表现之间的关系。方法采用巢式PCR法，对具有不同临床表现的122份HCMV感染患儿的尿标本和53份先天性巨结肠患儿痉挛段肠组织标本的临床株进行UL144开放阅读框的扩增，扩增阳性的临床株进行UL144开放阅读框核苷酸测序。结果50份尿标本和23份肠组织标本临床株完成DNA测序。种系进化树分析结果显示HCMVUL144基因分为3组4个基因型，UL144G1a（52％）为主要基因型。有症状与无症状HCMV感染临床株的基因型分布比较，差异无显著性。引起神经系统及肝胆系统受累的临床株与无症状感染临床株基因型分布比较，差异无显著性。与美国及日本的临床株比较，UL144基因型分布差异具有显著性。结论HCMVUL144基因具有高度多态性；UL144G1a为先天性或围生期HCMV感染的主要基因型；UL144基因分布与地理位置有关；UL144基因分型与HCMV感染的临床表现及组织嗜性无关。     

筛选与分析与人巨细胞病毒UL133编码蛋白相互作用的蛋白

目的 应用酵母双杂交技术筛选人胎脑cDNA文库中与人巨细胞病毒(HCMV)UL/b′ UL133编码蛋白相互作用的蛋白,为研究UL/b′编码蛋白的生物学功能,揭示HCMV先天感染的致病机制奠定基础.方法 设计特异性引物,在引物上下游分别引入NdeⅠ和BamHⅠ限制性内切酶识别位点,采用PCR技术扩增 HCMV临床分离株H株的UL133基因片段,将其扩增产物与载体pGBKT7 同时使用NdeⅠ和BamHⅠ进行双酶切,纯化后,将UL133片段插入到pGBKT7载体上,构建诱饵质粒 pGBKT7-UL133,并测序分析.利用醋酸锂小量酵母转化方法,将测序正确的诱饵质粒pGBKT7-UL133转化入AH109酵母感受态细胞中,然后将转化的菌液涂布在色氨酸缺陷型培养基(-Trp)上,筛选阳性菌落.再将人胎脑cDNA文库质粒转化到含pGBKT7-UL133质粒的AH109菌株中,在四缺培养基(-Ade/-His/-Leu/-Trp)中筛选,在铺有X-Gal的滤纸上进行显色反应,提取显蓝色的阳性酵母筛选质粒,运用电穿孔方法将其转化到TG1的大肠杆菌中,并行PCR鉴定,提取大肠杆菌中的文库质粒,并将其回转到含诱饵质粒pGBKT7-UL133 的酵母菌株中,进行回转验证.对筛选到的阳性克隆进行测序和生物信息学分析.结果 用于酵母双杂交筛选的诱饵质粒pGBKT7-UL133成功构建,含有诱饵质粒的酵母菌株在-Trp上生长.转化有人胎脑cDNA文库和诱饵质粒pGBKT7-UL133的酵母菌AH109,在四缺培养基中观察到有28个酵母菌落生长,通过显色反应、酵母质粒转化及回转酵母细胞筛选得到4个阳性克隆.通过序列比对,发现在这些基因中,其中一个基因编码人类还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)依赖性氧化还原酶1.结论 成功应用酵母双杂交系统筛选出与HCMV UL/b′区UL133编码蛋白相互作用的蛋白-NADPH,提示UL133蛋白可能在增加病毒毒力和传染性,干扰细胞间的信号传导和细胞的生长、分裂、凋亡等方面发挥重要作用.     

人巨细胞病毒UL146基因α趋化因子在巨细胞病毒感染中的作用

人巨细胞病毒(HCMV)在人群中感染非常普遍,对于免疫功能正常的人来说通常是不显性感染或潜伏感染,但胎儿及免疫缺陷患者感染可导致极高的病死率.目前,关于HCMV先天感染的致病机制还不十分清楚.HCMV UL146基因编码α趋化因子同源物,其核苷酸及氨基酸序列的变异和蛋白趋化因子功能区的高度保守提示这个功能区对于HCMV具有重要的生物学意义.     

人巨细胞病毒UL 146基因α趋化因子在巨细胞病毒感染中的作用

人巨细胞病毒（HCMV）在人群中感染非常普遍,对于免疫功能正常的人来说通常是不显性感染或潜伏感染,但胎儿及免疫缺陷患者感染可导致极高的病死率。目前,关于HCMV先天感染的致病机制还不十分清楚。HCMV UL146基因编码α趋化因子同源物,其核苷酸及氨基酸序列的变异和蛋白趋化因子功能区的高度保守提示这个功能区对于HCMV具有重要的生物学意义。     

人巨细胞病毒UL/b’区基因编码蛋白生物学功能的研究进展

人巨细胞病毒(human cytomegalovirus,HCMV)在人群中感染广泛,不仅是免疫抑制患者的严重致病原,也是引起新生儿先天及围生期感染最常见、危害最大的一种病原体.近年来,针对区别于实验室株的一个特殊区域UL/b’区基因编码蛋白生物学功能的研究是国内外HCMV研究领域的一个热点.目前已经发现该区域基因产物在HCMV的毒力、传播、组织细胞嗜性及免疫逃避等方面发挥重要作用,其中的UL133-UL138位点可能促进HCMV潜伏感染的建立和维持,而pUL138是目前病毒基因组中鉴定出的惟一明确与HCMV潜伏感染相关的决定因子.因此,HCMV UL/b’区基因产物在HCMV感染过程中发挥重要作用.该文就HCMV UL/b’区部分基因,尤其是与HCMV潜伏感染相关的UL138基因结构及其编码蛋白生物学功能进行综述,为阐明HCMV感染致病机制奠定基础.     

我国再障患儿血清中人细小病毒B19 NS基因序列测定及变异分析(英文)

目的：探讨B19病毒中国株NS基因的变异。方法：采用聚合酶链反应(PCR)技术,从中国再障患儿血清中扩增人细小病毒B19-XA17,XA18,XA20株的NS基因片段,进行核苷酸序列分析。结果：人细小病毒B19-XA17、XA18、XA20株的NS基因核苷酸序列被测定,测序结果与国外Au株基因核昔的序列比较,有4个核苷酸突变,并引起编码的4个氨基酸发生改变。结论：我国B19病毒NS基因序列与Au株相比有变异。     

幽门螺杆菌儿童分离株尿素酶基因B的克隆及序列分析

目的克隆幽门螺杆菌(Hp)临床儿童分离株尿素酶B(UreB)基因入pGEX-4T-1表达载体,并进行测序及基因比对分析,为以UreB作为Hp疫苗分子的口服疫苗研制奠定基础。方法根据GenBank中HpUreB序列,设计一对特异性引物PCR扩增Hp临床儿童分离株UreB全长基因,EcoRI及NotI酶切后与做相应酶切的pGEX-4T-1连接,转化大肠杆菌BL21,提取质粒进行双酶切鉴定及基因测序,并对测序结果进行比对分析。结果以Hp儿童分离株GZCH1为模板,成功扩增了UreB基因,基因大小为1710bp,重组pGEX-4T-1-Ure双酶切鉴定可见目的片段,测序结果显示UreB在正确阅读框中,序列比对分析显示其与相关报道序列核苷酸和氨基酸一致性达98%。Hp儿童分离株GZCH1UreB序列已登录GenBank(登录号:FJ455126)。结论从Hp儿童分离株GZCH1中成功克隆了UreB基因,为UreBHp口服疫苗研制奠定了基础。     

Allergic factors associated with the development of asthma and the influence of cetirizine in a double-blind, randomised, placebo-controlled trial: First results of ETA®

There is a common progression known as the allergic march from atopic dermatitis to allergic asthma. Cetirizine has several antiallergic properties that suggest a potential effect on the development of airway inflammation and asthma in infants with atopic dermatitis. Methods. Over a two year period, 817 infants aged one to two years who suffered from atopic dermatitis and with a history of atopic disease in a parent or sibling were included in the ETAC^® (Early Treatment of the Atopic Child) trial, a multi-country, double-blind, randomised, placebo-controlled trial. The infants were treated for 18 months with either cetirizine (0.25mg/ kg b.i.d.) or placebo. The number of infants who developed asthma was compared between the two groups. Clinical and biological assessments including analysis of total and specific IgE antibodies were performed. Results. In the placebo group, the relative risk (RR) for developing asthma was elevated in patients with a raised level of total IgE (≥ 30 kU/I) or specific IgE (≥ 0.35 kUA/I) for grass pollen, house dust mite or cat dander (RR between 1.4 and 1.7). Compared to placebo, cetirizine significantly reduced the incidence of asthma for patients sensitised to grass pollen (RR = 0.5) or to house dust mite (RR = 0.6). However, in the population that included all infants with normal and elevated total or specific IgE (intention-to-treat - ITT), there was no difference between the numbers of infants developing asthma while receiving cetirizine or placebo. The adverse events profile was similar in the two treatment groups. Discussion. Raised total IgE level and raised specific IgE levels to grass pollen, house dust mite or cat dander were predictive of subsequent asthma. Cetirizine halved the number of patients developing asthma in the subgroups sensitised to grass pollen or house dust mite (i.e. 20% of the study population). In view of the proven safety of the drug, we propose this treatment as a primary pharmacological intervention strategy to prevent the development of asthma in specifically sensitised infants with atopic dermatitis.     

Resurgence of measles in Singapore: profile of hospital cases

OBJECTIVE: To ascertain the profile of cases of measles seen at a general hospital during a recent outbreak that occurred despite a measles vaccination program. METHODOLOGY: A retrospective study from January 1991 to March 1998. All patients with measles (ICD code 055. 9) seen at the emergency unit or as inpatients were included. RESULTS: There were 87 cases identified. The diagnosis was clinical in all and proven serologically in 71%. Eighty-five per cent of the cases occurred between January 1997 and March 1998. There was a bi-modal age distribution with peaks in the very young (相似文献   

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孤独症谱系障碍(autistic-spectrum disorders,ASDs)近年来患病率逐年攀升至1%左右,其症状往往伴随终生,成为严重威胁儿童健康和发展的神经发育性疾患;注意缺陷多动障碍(attention deficit hyperactivity disorder,ADHD)是儿童期最常见的精神障碍,国内报道患病率为4.13%～5.83%,其症状可延续至青少年期,甚至到成年期[1]。这两类精神障碍在成年期的临床表现、共患病、治疗策略和预后与儿童期有哪些不同呢?本文通过回顾相     

EXCITING NEW TREATMENT APPROACHES FOR PATHYPHYSIOLOGIC MECHANISMS OF SICKLE CELL DISEASE

During the past several decades, our understanding of the complex pathophysiology of vasoocclusion associated with sickle cell disease has improved greatly. Interaction of genes, hemoglobin molecules, red cell membrane and metabolic changes, cell-cell interactions and cell-plasma interactions, red cell adhesion to vascular endothelium, activation of coagulation, and vascular reactivity play a role in vaso occlusion. Penicillin prophylaxis of pneumococcal infections and appropriate use of blood transfusions and other supportive measures improved survival of sickle cell patients. Hydroxyurea made a major impact on sickle cell therapy when it was shown to decrease acute painful episodes, acute chest syndrome, and the need for blood transfusion in adults. Significant experience in the use of hydroxyurea has been accumulated in older children. The benefits and risks of hydroxyurea for younger children and long-term risks in all patients will be evaluated in future investigations. Other promising therapies include butyrate compounds, clotrimazole, magnesium supplementation, poloxamer 188, antiadhesion agents, anticoagulant approaches, and nitric oxide. Hemopoietic transplantation remains the only curative therapy. However, several transgenic mouse models are available for studies of gene therapy or other treatment approaches on biochemical, cellular, and pathologic effects of mutant genes.     

Infiltrations of Epstein-Barr virus-carrying cells in granular lymphocyte-proliferative disorders corresponding to chronic active Epstein-Barr virus infection

A 21-year-old man with granular lymphocyte-proliferative disorders (GLPD) associated with chronic active Epstein-Barr virus (EBV) infection is described. Chromosomal analyses revealed several clonal abnormalities and two of them were mainly repetitious. High copy numbers of monoclonal EBV genome were also detected in the proliferative large granular lymphocytes (LGLs), indicating the monoclonal expansion of EBV-infected LGLs. The patient had an indolent course for several years, and there was no evidence of infiltrations of his bone marrow until the end stage. At autopsy, microscopic studies revealed marked infiltrations of LGL in the liver and spleen, and the infiltrating cells were NK-cell immunophenotype. The infiltrated LGLs showed latency I.     

LUTEINIZING HORMONE RECEPTOR MUTATIONS IN DISORDERS OF SEXUAL DEVELOPMENT AND CANCER

Human male sexual development is regulated by chorionic gonadotropin (CG) and luteinizing hormone (LH). Aberrant sexual development caused by both activating and inactivating mutations of the human luteinizing hormone receptor (LHR) have been described. All known activating mutations of the LHR are missense mutations caused by single base substitution. The most common activating mutation is the replacement of Asp-578 by Gly due to the substitution of A by G at nucleotide position 1733. All activating mutations are present in exon 11 which encodes the transmembrane domain of the receptor. Constitutive activity of the LHR causes LH releasing hormone-independent precocious puberty in boys and the autosomal dominant disorder familial male-limited precocious puberty (FMPP). Both germline and somatic activating mutations of the LHR have been found in patients with testicular tumors. Activating mutations have no effect on females. The molecular genetics of the inactivating mutations of the LHR are more variable and include single base substitution, partial gene deletion, and insertion. These mutations are not localized and are present in both the extracellular and transmembrane domain of the receptor. Inactivation of the LHR gives rise to the autosomal recessive disorder Leydig cell hypoplasia (LCH) and male hypogonadism or male pseudohermaphroditism. Severity of the clinical phenotype in LCH patients correlates with the amount of residual activity of the mutated receptor. Females are less affected by inactivating mutation of the LHR. Symptoms caused by homozygous inactivating mutation of the LHR include polycystic ovaries and primary amenorrhea.     

A profile of respiratory symptoms in urban and rural South Australian school children

This report describes the cross-sectional analyses of data from the first year of a longitudinal study using questionnaire and respiratory function data over a 5 year period from a sample of rural South Australian school children. The cumulative or lifetime prevalences of respiratory symptoms were estimated in 825 rural and 1261 urban school children aged between 5 and 15 years in order to determine if the prevalence rates differed between rural and urban school children. The study found the overall cumulative prevalence of asthma and/or wheezy breathing (AWB) to be 24.1% in the rural school children compared to 27.6% in the urban school children. Most children developed AWB symptoms before the age of 7 years, with 20% reporting moderately severe symptoms and 10% having more than one attack per fortnight. The cumulative prevalence of bronchitis, loose/rattly cough (BLRC) differed significantly between the rural school children (34.1%) and urban school children (47.9%). The BLRC symptoms preceded the development of AWB in many cases. Urban school children also reported a higher prevalence of atopic conditions.     

Personality profiles and heart rate variability (vagal tone) in children with recurrent abdominal pain

The aim of the study was to explore psychological factors and autonomic activity in children with recurrent abdominal pain and to compare them with those in a control group of healthy children. The Personality Inventory for Children was used for assessment of developmental, emotional and psychosocial factors in 25 children with recurrent abdominal pain (age, 7-15 y). Parasympathetic and sympathetic functions in these children and in 23 healthy control subjects (age, 7-13 y) were also investigated, non-invasively using a computerized polygraph. Vagal tone (parasympathetic function) was indexed by calculation of respiratory sinus arrhythmia in beats/min. Skin conductance (sympathetic function) was recorded by the constant current method. On the Personality Inventory for Children, 16 patients had high scores on somatic concern. Several patients had scores in the clinical range for depression, withdrawal and anxiety, but the mean scores for these personality profile scales were well within the normal range of healthy children. Interestingly, there was a spike on the L (Lie)-scale for most of the patients and 15 patients had scores above or close to the clinical cut-off value. As compared with the scores in healthy children, vagal tone and sympathetic tone were normal. Conclusion: Many children with recurrent abdominal pain have scores in the clinical range for depression, withdrawal, anxiety and L-scale indicating coping problems, denial and a trend towards somatic concern that may contribute to the evolution of abdominal pain. Autonomic nerve activity was not disturbed in these children.     

Hauttemperaturen verschiedener Körperoberflächenareale des Rumpfes bei Säuglingen

Summary In two groups of infants (3–53 weeks old) skin temperatures were controlled in different areas of the trunk—i.e.: regions of sternum, lungs, heart, liver, spleen, kidneys—at different room-temperatures (group I: 21–25°C; group II: 29–32°C). Rectal temperatures of some probands in both groups also had been controlled simultaneously. A definite change in the reaction to heat was proofed in different periods of the first year of life. In higher environmental temperatures the skin temperature was almost constant at every controll-point of the skin, even in older infants. In lower environmental temperatures the skin temperatures lowered continuously with age till 7. to 9. moth. From 10. to 12. month the lowering of skin temperature discontinued. The rectal temperatures were relatively constant in all infants. Only in infants from 7. to 12. month, whose skin temperatures were controlled in lower as well as in higher environmental temperatures, a tendency to higher rectal temperatures was proofed in warmer environmental temperatures.The significance of these results is discussed.

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Untersuchungen mit Unterstützung durch die Deutsche Forschungsgemeinschaft.