Genotyping of Mycoplasma pneumoniae isolates using real-time PCR and high-resolution melt analysis

Mycoplasma pneumoniae is an important respiratory pathogen, accounting for up to 25% of community-acquired pneumonia, and is a common cause of hospitalized pneumonia in otherwise healthy adults and children. Mycoplasma pneumoniae isolates can be classified into two main genomic groups (type 1 and type 2) based on sequence variation within the gene encoding the major adhesion molecule P1. Although numerous publications have described real-time PCR assays for the detection of M. pneumoniae , none has been able to discriminate the two genomic types. Here, a real-time PCR assay that can distinguish each type of M. pneumoniae utilizing high-resolution melt-curve analysis is reported. Using this method, 102 isolates obtained from patients from 1965 to the present, including those from recent outbreaks, were typed along with reference strains M129 (type 1) and FH (type 2). The results show that 55 isolates (54%) can be classified as type 1 and 47 isolates (46%) as type 2, and 100% correlation was demonstrated when compared with a standard PCR-restriction fragment length polymorphism typing procedure. Typing of isolates obtained from recent outbreaks in the USA has revealed the presence of both types. This assay provides a rapid, reliable and convenient method for typing M. pneumoniae isolates and may be useful for surveillance purposes and epidemiological investigations, and may provide insight into the biology of M. pneumoniae distribution within populations.     

Molecular biology of mantle cell lymphoma: From profiling studies to new therapeutic strategies

Mantle cell lymphoma (MCL) is a well-defined lymphoid malignancy characterized by a rapid clinical evolution and poor response to current therapeutic protocols. The hallmark genetic alteration of MCL is the t(11;14)(q13;32) chromosomal translocation that leads to the overexpression of cyclin D1. Recently, new molecular alterations of major importance in the pathogenic mechanisms of this disease have been discovered, and have revealed the biological heterogeneity of MCL. The first section of our review discusses our current understanding of the molecular biology of this entity according to recent information from comparative genomic hybridization (CGH) and expression profiling studies, which are leading to the identification of several druggable targets. In the second section we revise new therapeutic strategies based on new drug families that target key molecular pathways of major relevance in this malignancy. We analyze emerging agents that are already producing significant results in different models of human cancers, including MCL. Based on the current knowledge and recent studies, we suggest that the encouraging results described here should provide a rationale platform for the design of new treatments that may overcome the resistance of this aggressive lymphoma to conventional therapy and improve patient prognosis.     

Up-regulation of microtubule-associated protein 2 accompanying the filial imprinting of domestic chicks (Gallus gallus domesticus)

Using cDNA microarrays, we have identified elsewhere the genes of microtubule-associated proteins as a group up-regulated in newly hatched chick brains after filial imprinting training. Here we show by in situ hybridization that the mRNA for the microtubule-associated protein 2 (MAP2) gene was enriched in the mesopallium and the hippocampus in the trained chick brain. The regionally specific enrichments of MAP2 mRNA were not observed in the brain of dark-reared or light-exposed chick as controls, implying an association between the degree of expression and the strength of the learned preference. In agreement with the gene expression, MAP2 protein was accumulated in the mesopallium of the trained chick brain, but not in the brains of the controls. The accumulation of MAP2 was found in the cytosol of neurons and co-localized with β-tubulin, suggesting a change in microtubule assembly. Our results suggest a postnatal reorganization of cytoskeleton following filial imprinting.     

Effect of perinatal stress on the biogenic amine neurotransmitter level of the adult rat's brain

The amount of biogenic amines (dopamine and serotonin) and their metabolites (DOPAC, HVA, 5-HIAA, and 5-HTOL) in five regions of the brain (frontal cortex, hypothalamus, hippocampus, striatum, and brainstem) was studied in the male and female offspring of control and perinatally (48 h before birth or 48 h after birth) food and water deprived dams, when they were three months old, by using HPLC-EC determination. The increase of amine or metabolite level was dominant (19 values increased and 10 decreased related to control). Before-birth stress caused increase in 9 case and only 2 decreased, while in the case of after-birth stress 10 increased and 8 decreased. However, though there is no possibility to decide an exact tendency of direction, the after-birth stress (transmitted by milk) has more expressed effect. Striatum and brainstem were the most touched regions. There was a gender dependence with the dominance of males, except striatum. Blood plasma nociceptin level was also studied and there was a significant elevation in males after pre- and postnatal deprivation, while in females only after postnatal deprivation. The importance of the results in correlation with other stress effects is discussed.     

应用聚合酶链反应检测日本血吸虫基因组DNA

中国是日本血吸虫病主要流行区[1],疫情形势严峻.准确评估钉螺感染和水体污染情况,对疫情防制意义重大.传统的压碎法、逸蚴法和哨鼠法灵敏性都较低[2],影响了血防工作的效率.有人尝试将PCR方法用于血吸虫病的诊断和流行监测[3-5],其中根据埃及和曼氏血吸虫中发现的串联重复序列建立的PCR方法都表现出极高的灵敏性[6-8],实用前景乐观[9].本研究的主要目的是揭示日本血吸虫中可能存在的该序列,优化PCR条件,评价其灵敏性.     

Gadd45与基因组稳定性

Gadd45基因是生长抑制及DNA损伤诱导基因家族中的一员,是电离辐射效应基因之一,在细胞周期调控、DNA损伤修复及细胞凋亡过程中发挥重要作用,这使它成为维持基因组稳定性的重要基因,在肿瘤细胞存活、凋亡信号通路中发挥错综复杂的作用,从而参与肿瘤的发生和发展.     

分子印迹技术原理及其在药物分析检测中的应用

分子印迹技束是20世纪90年代快速发展起来的。制备空间结构和结合位点与模板分子完全匹配的聚合物的技术,它具有构效预定性、特异识别性和广泛实用性的特点。本文主要介绍分子印迹技术的产生、原理及其在药物分析检测中的应用．并对此项新技术的未来作了展望。     

中国乙型肝炎病毒基因组序列突变及进化分析

目的 了解中国乙型肝炎病毒（hepatitis B virus,HBV）基因组序列的免疫逃逸突变、耐药突变和基因组进化信息。方法 选取1998-2021年在GenBank中的中国HBV全基因组序列信息作为分析对象,采用MAFFT软件进行聚类分析,使用在线工具Gen2pheno进行免疫逃逸突变和耐药突变分析,使用BEAST 1.10.4做序列的时间进化分析。结果 5 426条序列纳入分析集,分布于我国19个省份,C型占比最高（59.1%,3 211/5 426）。其次为B型（33.7%,1 833/5 426）。有764条（14.1%,764/5 426）序列发生了免疫逃逸突变,98.1%的序列至少发生了1处反转录酶编码区域突变。中国大部分系列的进化根在公元1801年左右。结论 中国HBV耐药突变率较高,HBV基因组进化缓慢。     

New Insights into the Genetics of Neonatal Diabetes

Reviews in Endocrine and Metabolic Disorders -