Molecular epidemiological study of HEV-B enteroviruses involved in the increase in meningitis cases occurred in Spain during 2006

Human enteroviruses are one of the main etiological agents of aseptic meningitis and other central nervous system infections, particularly the serotypes included in the enterovirus B species. Molecular methods have proved useful to identify serotypes in clinical samples, facilitating the epidemiological study of these viruses. In the spring of 2006, there was a significant increase in meningitis cases caused by enteroviruses in Spain. In the present study, 138 enteroviruses directly detected in clinical samples of patients with aseptic meningitis (n = 116) and other neurological pathologies (n = 22) received by the National Center for Microbiology during the year, were genotyped by amplification and sequencing part of the VP1 region and phylogenetic analysis. Echovirus 30 was the most frequent serotype, followed in decreasing order by echovirus 6, 9, 13, 18, enterovirus 75, coxsackievirus A9, echovirus 11, 14, 29, 4, and coxsackievirus B4 and B5. Phylogenetic analysis with all Spanish echovirus 30 strains detected in 2006 and other reported echovirus 30 sequences, demonstrated that Spanish strains formed a new lineage, different from others previously described. In conclusion, echovirus 30 is the most commonly reported enterovirus serotype associated with aseptic meningitis in Spain. Direct molecular typing of clinical samples also allows rapid identification of the serotypes involved in an epidemic alert and phylogenetic analysis in the 3'-VP1 region is useful to study viral epidemiology.     

Influence of immunosuppressive drugs on the CD30 molecule in kidney transplanted patients

Background

Soluble CD30 (sCD30) is a suggested marker for kidney transplantation outcomes. We investigated whether sCD30 serum levels are influenced by immunosuppression and whether they correlate with findings in protocol biopsies and with CD30 gene expression in peripheral blood mononuclear cells (PBMC).

Mir-30a抑制自噬及上皮细胞间质化增强肺腺癌细胞对克唑替尼的敏感性

目的探索上调肺腺癌细胞中Mir-30a的表达水平能否增强细胞对克唑替尼的敏感性,观察细胞增殖率的变化及克唑替尼IC50等的变化,并探索相关的机制。方法使用Lipofectamine2000携带Mir-30a对H3122细胞进行瞬时转染,并与克唑替尼共同作用。将肺腺癌H3122细胞分为对照组、克唑替尼与联合组。MTT法检测各组细胞增殖率变化,Transwell检测细胞侵袭性。Western blot检测ALK、c-MET、Beclin-1、E-cadherin及Vimentin蛋白的表达。结果在H3122细胞中Mir-30a转染与克唑替尼共同作用较克唑替尼单独作用有更显著的细胞杀伤作用。转染后的细胞较单独应用克唑替尼组侵袭力减弱。转染与克唑替尼联合组Beclin1、ALK、c-MET及Vimentin的蛋白的表达有明显下降,E-cadherin稍有增强。结论 Mir-30a增强肺腺癌细胞对克唑替尼的敏感性,可能与过量表达的Mir-30a抑制癌细胞的自噬及上皮细胞间质化相关。     

miR-30a-3p靶向NOD1对心肌细胞缺氧复氧损伤的影响

目的　探究miR-30a-3p与NOD1的靶向关系,及其对心肌细胞缺氧复氧损伤的影响和机制。 方法　将细胞分为Ctrl组、H/R组、miR-30a-3p mimic组和miR-30a-3p inhibitor组,经过缺氧复氧处理后,转染相应的miRNA处理细胞,RT-PCR检测miR-30a-3p和NOD1基因表达水平,荧光素酶报告检测miR-30a-3p和NOD1靶向关系,Western blot检测NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平,CCK8法检测细胞活性,Hoechst检测细胞凋亡,试剂盒检测LDH、CK、MDA、T-SOD水平。 结果　miR-30a-3p表达水平随缺氧复氧处理时间增长而下降,NOD1基因表达水平随缺氧复氧处理时间增长而升高。在荧光素酶报告实验中,NOD1 WT+miR-30a-3p mimic荧光素酶活性显著低于NOD1 WT+NC组。与Ctrl组比较,H/R组miR-30a-3p基因表达水平、细胞活性、T-SOD水平降低,NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率升高;与H/R组比较,miR-30a-3p mimic组miR-30a-3p基因表达水平、细胞活性、T-SOD水平升高,NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率降低,miR-30a-3p inhibitor组miR-30a-3p基因表达水平、细胞活性、T-SOD水平降低,NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率升高。 结论　miR-30a-3p可靶向作用于NOD1,缓解心肌细胞缺氧复氧造成的损伤,其作用机制可能与调控NF-κB信号通路有关。     

Transmembrane G protein-coupled receptor 30 gene polymorphisms and uterine adenomyosis in Korean women

Abstract

To compare the genetic distributions of 14G protein-coupled receptor 30 (GPR30) single-nucleotide polymorphisms (SNPs) between women with and without uterine adenomyosis. The study population comprised 69 Korean women. Uterine tissues from the adenomyosis and non-adenomyosis groups were used for DNA extraction. Pre-designed PCR/Sanger or Sequencing Primer and TaqMan^® SNP Genotyping Assays were used for the SNP genotyping of the GPR30 gene. Immunohistochemical staining was performed to confirm the GPR30 expression. Differences in genotype and allele frequencies between the two groups were calculated using Fisher’s exact test. The rs3802141 CT genotype was more common in the control group (p?=?.02), and the rs4266553?CC genotype was more common in the adenomyosis group (p?=?.02). The C allele of the SNP rs4266553 was more common in the adenomyosis group (p?=?.02). GPR30 expression was confirmed in 69 individuals in both groups. GPR30 gene polymorphism is presumed to affect the risk of adenomyosis with limited sample size. Further large-scale study is needed to explain the genetic influence of GPR30 gene polymorphism.     

miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤细胞增殖

目的 探讨miR-30-5p对视网膜母细胞瘤细胞增殖的影响及作用机制。 方法 采用实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-30-5p在视网膜母细胞瘤细胞系和人视网膜内皮细胞(HRECs)中的表达。利用TargetScan数据库进行生物信息学分析并预测miR-30-5p靶基因,双荧光素酶报告基因实验验证miR-30-5p与FOXG1的3'UTR结合能力及靶向关系,qRT-PCR和Western blotting分别检测miR-30-5p对FOXG1的mRNA与蛋白表达影响。瞬时转染Y79细胞后,通过CCK8法检测各组Y79细胞的增殖。 结果 与HRECs比较,miR-30-5p在视网膜母细胞瘤细胞中表达降低,FOXG1在视网膜母细胞瘤细胞中表达升高。生物信息学预测结果显示miR-30-5p与FOXG1存在结合位点,qRT-PCR和Western blotting显示miR-30-5p可负调控FOXG1的mRNA和蛋白的表达,双荧光素酶实验结果证实miR-30-5p与FOXG1 3'UTR存在靶向关系。瞬时转染miR-30-5p可升高Y79细胞中miR-30-5p的表达水平,抑制Y79细胞的增殖活力。过表达FOXG1可逆转miR-30-5p上调对Y79细胞增殖的影响。 结论 miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤Y79细胞的增殖。     

CD30阳性的窦性大B细胞淋巴瘤临床病理学分析

目的 探讨CD30阳性的窦性大B细胞淋巴瘤（SLBCL）的临床病理学特征及其与EpsteiBarr(EB)病毒的关系。方法 结合临床特点、形态学及免疫表型分析,并通过聚合酶链反应（PCR）法检测EB病毒BamHIW DNA片段及免疫组织化学法检测潜肽膜蛋白1（LMP1）,对2例CD30阳性的SLBCL进行分析。结果 2例分别为64岁男性及85岁女性,均表现为浅表淋巴结肿大,其中1例10个月后死于肿瘤。镜下,肿瘤都以窦性分布且彼此黏附的生长方式及相对单形的细胞形态为特征。免疫表型分析提示,两者均呈CD45、CD30、CD20或CD79α阳性而不表达上皮膜抗原（EMA）、ALK1及组织细胞或T细胞的标志。未发现2例有EB病毒感染的证据。结论 CD30阳性的SLBCL是一种形态、表型独特的弥漫大B细胞淋巴瘤变异类型,该瘤需与T/裸细胞性间变性大细胞淋巴瘤及其他一些累及淋巴窦为主的淋巴结病变相鉴别,但与EB病毒可能无关。     

Graves病患者治疗前后血清CD30水平的观察

<正> Graves病(GD)是一种器官特异性自身免疫性疾病,其特点是甲状腺内有淋巴细胞的浸润,疾病活动期血循环中存在激活的T细胞和自身抗体,然而,具体的发病机制不详。本文通过对GD患者治疗前后血清CD30水平的测定,探讨CD30分子在GD发病机制中的作用。     

Predominance of enterovirus B and echovirus 30 as cause of viral meningitis in a UK population

Background/ObjectivesEnteroviruses are the most common cause of aseptic or lymphocytic meningitis, particularly in children. With reports of unusually severe neurological disease in some patients infected with enterovirus D68 in North America, and a recent increase in the number of paediatric enterovirus meningitis cases presenting in this UK Midlands population, a retrospective regional surveillance study was performed.Study designCerebrospinal fluid (CSF) samples received were tested using the polymerase chain reaction (PCR) for HSV-1/2, VZV, enteroviruses and parechoviruses. Enterovirus PCR positive CSF samples were sent for further serotyping. A phylogenetic tree was constructed of the echovirus 30 VP1 sequences, where sufficient sample remained for sequencing.ResultsThe number of enterovirus positive CSFs from each year were: 21 (2008), 7 (2011), 53 (2012), 58 (2013) and 31 (2014). Overall, 163 of the 170 serotyped enteroviruses belonged to the species B (echovirus 5, 6, 7, 9, 11, 13, 16, 17, 18, 21, 25, 30; coxsackie B1, B2, B3, B4, B5, A9), with only 7 belonging to species A (coxsackie A2, A6, A16 and enterovirus 71). Echovirus 30 was the predominant serotype overall, identified in 43 (25.3%) of samples, with a significantly higher proportion in the adult age group (37.3%) compared to the infant age group (12.3%). Phylogenetic analysis showed that these UK Midlands echovirus 30 VP1 sequences clustered most closely with those from Europe and China.ConclusionThis study showed a continued predominance of echovirus 30 as a cause of viral meningitis, particularly in adults, though more surveillance is needed.