胰岛素样生长因子1对早孕滋养细胞增殖的作用及机制研究

目的探讨胰岛素样生长因子1(IGF-1)对体外培养的早孕滋养细胞增殖的影响,以及滋养细胞增殖的细胞内信号转导机制。方法取原代培养传代后生长良好的滋养层细胞,加入不同浓度的IGF-1继续培养,用四唑盐(MTT)比色法测定细胞的增殖活性,并且以ERK(细胞外信号调节蛋白激酶)通路的特异性抑制剂U0126处理细胞,间接反映ERK通路的作用。结果①与对照组相比,IGF-1浓度≥1nM时,其促进滋养细胞增殖效果有显著性意义(P<0.05),且在0.1-100nM范围内,此种作用与浓度呈正相关。②ERK通路阻滞剂U0126可抑制滋养细胞的增殖(P<0.05),并且可显著抑制IGF-1对滋养细胞的促增殖作用(P<0.05)。结论①IGF-1对滋养层细胞的增殖活性具有促进作用。②ERK通路可能是滋养细胞增殖过程中的主要信号转导通路之一。     

丹参酮ⅡA通过PI3K/AKT/ERK信号通路对脑胶质瘤细胞增殖、凋亡、侵袭及迁移的影响北大核心CSCD

目的:探讨丹参酮ⅡA通过磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)/细胞外调节激酶(ERK)信号通路对脑胶质瘤细胞增殖、凋亡、侵袭及迁移的影响。方法:取对数生长期的U251细胞,并分为空白组(未处理的U251细胞)、对照组(0.1%DMSO)、丹参酮ⅡA组(20μmol/L丹参酮ⅡA)、IGF-1组(100 ng/ml PI3K/AKT/ERK通路激活剂IGF-1)、丹参酮ⅡA+IGF-1组(20μmol/L丹参酮ⅡA与100 ng/ml IGF-1同时处理),倒置显微镜观察各组细胞形态;CCK-8法检测各组细胞增殖;流式细胞术检测各组细胞凋亡;Transwell实验检测各组细胞侵袭与迁移;Western blot检测各组细胞中细胞周期素D1(CyclinD1)、B淋巴细胞瘤-2相关蛋白(Bax)、基质金属蛋白酶2(MMP-2)、MMP-9及PI3K/AKT/ERK通路相关蛋白的表达。结果:与空白组、对照组比较,丹参酮ⅡA组U251细胞数量减少,细胞皱缩,出现大量细胞碎片,而IGF-1组U251细胞数量增多,细胞形态未发生改变;与丹参酮ⅡA组比较,丹参酮ⅡA+IGF-1组细胞数量增多,细胞贴壁性变强。与空白组、对照组比较,丹参酮ⅡA组U251细胞OD450值(24 h、48 h)、侵袭细胞数目、迁移细胞数目、CyclinD1、MMP-2、MMP-9蛋白表达水平显著降低,细胞凋亡率、Bax蛋白表达水平显著升高(P<0.05),而IGF-1组U251细胞OD450值(24 h、48 h)、侵袭细胞数目、迁移细胞数目、CyclinD1、MMP-2、MMP-9蛋白表达水平显著升高,细胞凋亡率、Bax蛋白表达水平显著降低(P<0.05);与丹参酮ⅡA组比较,丹参酮ⅡA+IGF-1组U251细胞OD450值(24 h、48 h)、侵袭细胞数目、迁移细胞数目、CyclinD1、MMP-2、MMP-9蛋白表达水平显著升高,细胞凋亡率、Bax蛋白表达水平显著降低(P<0.05)。结论:丹参酮ⅡA通过抑制PI3K/AKT/ERK通路抑制U251细胞增殖、侵袭、迁移,并诱导其凋亡。     

ERK1/2MAPK通路在Eca109细胞体外增殖和迁移中的作用及其机制

目的观察ERK1/2 MAPK通路在食管鳞状细胞癌(ESCC)Eca109细胞系增殖和迁移中的作用并探讨其作用机制。方法使用MAPK(ERK1/2)抑制剂U0126处理Eca109细胞;细胞计数检测细胞增殖;倒置显微镜下观察细胞形态;实时荧光定量PCR(qRT-PCR)检测ERK1/2 mRNA;Western blot检测总ERK1/2(t-ERK1/2)和磷酸化ERK1/2(p-ERK1/2);MTT和细胞划痕实验检测细胞增殖和迁移能力;qRT-PCR检测MicroRNA-21(miR-21)。结果20μmol/L浓度的U0126可抑制Eca109细胞生长(P<0.05),破坏细胞正常形态,ERK1/2 MAPK通路的活化在蛋白质水平被抑制(P<0.05),Eca109细胞增殖和迁移明显减弱(P<0.05),显著下调miR-21的表达(P<0.05)。结论ERK1/2 MAPK信号通路抑制可减弱Eca109细胞增殖和迁移,其可能的作用机制之一与其抑制miR-21表达有关。     

人参皂苷Rb1对体外培养胎鼠神经干细胞增殖及分化的影响

目的:探讨人参皂苷Rb1对体外培养胎鼠神经干细胞(NSCs)增殖和分化的影响及其可能的作用机制。方法:分离培养胎鼠皮层NSCs,CCK-8法及BrdU掺入法测定不同浓度人参皂苷Rb1对NSCs增殖的影响,确定促进NSCs增殖的最佳人参皂苷Rb1浓度;细胞免疫荧光法检测各组β-Ⅲtubulin、GFAP及DAPI的表达,计算GFAP/DAPI、β-Ⅲtubulin/DAPI百分比;Western Blot、CCK-8法测定PI3K/Akt抑制剂LY294002干预下Akt、磷酸化Akt(p-Akt)蛋白表达水平及其OD值。结果:人参皂苷Rb1能够促进体外培养的NSCs增殖,最佳浓度为1μmol/L;不同浓度人参皂苷Rb1组GFAP/DAPI及β-Ⅲtubulin/DAPI百分比与对照组相比无显著性差异(P0.05);但人参皂苷Rb1组(1μmol/L)的OD值及p-Akt蛋白水平均较对照组高(P0.01),且这种作用可以被LY294002逆转。结论:人参皂苷Rb1在一定浓度范围内可以促进体外培养神经干细胞的增殖,PI3K/Akt信号通路在人参皂苷Rb1促进神经干细胞增殖作用中发挥重要作用。     

细菌脂多糖通过MAPK / ERK1 / 2 信号通路影响成骨细胞活性和凋亡

目的:探讨细菌脂多糖(LPS)通过丝裂原活化蛋白激酶/细胞外调节蛋白激酶(MAPK/ERK1/2)信号通路对成骨细胞活性和凋亡的影响。方法:将小鼠前体成骨细胞MC3T3-E1随机分为对照组、LPS组、LPS+U0126组,对照组不给予LPS处理,LPS组给予10 mmol/L LPS处理,LPS+U0126组给予10 mmol/L LPS和25μmol/L ERK1/2激酶抑制剂U0126处理。采用噻唑蓝(MTT)测定细胞活性,采用流式细胞术测定细胞凋亡,采用硝基苯磷酸二钠基质动力学方法测定碱性磷酸酶(ALP),采用放射免疫法测定骨钙素(BGP)活性,采用Hochest 33258染色观察细胞核形态,采用Western blot测定活化型细胞半胱氨酸天冬氨酸特异性蛋白酶3型(C-Caspase 3)、Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2基因(Bcl-2)、ERK1/2、磷酸化ERK1/2(p-ERK1/2)蛋白水平。结果:与对照组比较,LPS组和LPS+U0126组MC3T3-E1细胞活性、ALP和BGP活性降低(P0.05),细胞凋亡率升高(P0.05),C-Caspase 3、Bax、p-ERK1/2蛋白水平升高(P0.05),Bcl-2蛋白水平降低(P0.05);与LPS组比较,LPS+U0126组MC3T3-E1细胞活性、ALP和BGP活性升高(P0.05),细胞凋亡率降低(P0.05),C-Caspase 3、Bax、p-ERK1/2蛋白水平降低(P0.05),Bcl-2蛋白水平升高(P0.05)。对照组细胞核正常;LPS组细胞凋亡形态数量明显增多;LPS+U0126组凋亡形态细胞数量较LPS组减少。结论:LPS可通过抑制MAPK/ERK1/2信号通路抑制成骨细胞活性,诱导成骨细胞凋亡。     

Erk信号途径在B104 CM诱导神经干细胞向少突胶质细胞前体分化中的作用

目的探讨细胞外信号调节激酶(Erk)信号途径及下游转录因子cf-os、cj-un等在神经母细胞瘤B104细胞系来源的条件培养基(B104 CM)诱导神经干细胞(NSCs)向少突胶质细胞前体(OPCs)分化中的作用。方法从形态学上观察Erk1/2特异性抑制剂U0126阻断对B104 CM诱导NSCs向OPCs分化的影响;分别以Western blotting和RT-PCR法检测对照组、B104 CM诱导组和U0126预孵组NSCs中Erk的磷酸化和转录因子cf-os、cj-un、c-myc的表达情况。结果U0126预孵可阻断B104 CM诱导的NSCs向OPCs分化;B104 CM可引起NSCs中Erk1/2迅速磷酸化和cf-os、cj-unmRNA表达上调,该作用可被U0126阻断。结论B104 CM通过活化Erk信号途径及其随后上调转录因子cf-os、cj-un的表达诱导NSCs向OPCs分化。     

ERK通路激动剂EGF及抑制剂U0126对HepG2肝癌细胞迁移和侵袭能力的影响

目的研究丝裂原活化蛋白激酶(MAPK)亚族细胞外调节蛋白激酶(ERK)通路在HepG2肝癌细胞迁移和侵袭中的作用。方法体外培养人肝癌HepG2细胞,采用ERK通路激动剂和抑制剂:表皮生长因子(EGF)及U0126刺激48h,分别应用MTS法、划痕实验法及Transwell小室法检测EGF和U0126对HepG2细胞增殖能力、迁移能力和侵袭能力的影响,qPCR和Western blot法分别检测ERK mRNA和磷酸化ERK蛋白的表达变化。结果细胞增殖、迁移和侵袭实验结果均显示,激动剂EGF可显著提高HepG2肝癌细胞的增殖、迁移及侵袭能力(P0.01),而U0126均对以上指标有抑制效应(P0.05)。qPCR和Western blot结果显示,EGF可上调ERK mRNA和磷酸化蛋白的表达(P0.01),而U0126对ERK mRNA无明显影响(P0.05),但可显著下调ERK磷酸化蛋白的表达(P0.01)。结论 ERK通路参与HepG2细胞的增殖、迁移和侵袭过程,EGF和U0126对以上指标的影响可能与ERK mRNA的表达及蛋白磷酸化过程相关。     

IL-33通过ERK1/2信号通路促进哮喘模型小鼠气道重塑

目的探讨白细胞介素(IL-33)诱导支气管哮喘气道重塑的机制。方法雄性BALB/c小鼠随机分为对照组、卵清蛋白(OVA)组和IL-33中和抗体联合OVA组。HE染色观察小鼠气道重塑,免疫组织化学染色及Western blot法观察IL-33、α平滑肌肌动蛋白(α-SMA)、1型胶原蛋白(Col1)的表达,Western blot法检测细胞外信号调节激酶1/2(ERK1/2)及丝裂原和应激激活的蛋白激酶1(MSK1)的磷酸化水平;培养HLF-1人成纤维细胞,分别给予人重组IL-33(r IL-33)、ERK1/2的抑制剂U0126联合r IL-33、MSK1的抑制剂H89联合r IL-33处理;实时荧光定量PCR及Western blot法检测α-SMA与Col1的mRNA和蛋白表达水平,免疫荧光细胞化学技术观察ERK1/2及MSK1的磷酸化。结果 OVA组小鼠发生气道重塑,IL-33、α-SMA、Col1表达增加,ERK1/2及MSK1磷酸化增强;IL-33中和抗体预处理可显著降低OVA诱导的小鼠气道重塑及IL-33、α-SMA、Col1表达以及ERK1/2及MSK1的磷酸化。U0126或H89可抑制r IL-33引起的HLF-1细胞中ERK1/2及MSK1磷酸化增强及α-SMA与Col1表达增加。结论 IL-33通过ERK1/2-MSK1信号通路促进哮喘模型小鼠气道重塑。     

18α-GA激活Nrf2对成体神经干细胞的影响

目的:明确18α-甘草次酸(18α-GA)对核因子E2相关因子2(Nrf2)的激活作用,观察上调Nrf2对成体神经干细胞(a NSCs)增殖和分化能力的影响,探讨维持a NSCs活力的有效途径。方法:体外培养新生、成年和老年小鼠的侧脑室室膜下区神经干细胞(NSCs),观察随年龄增长NSCs中Nrf2的表达变化。以18α-GA作用a NSCs,用real-time PCR和Western blot实验比较18α-GA组与DMSO对照组a NSCs的Nrf2表达变化。构建携带绿色荧光蛋白(GFP)基因的Nrf2-shRNA慢病毒载体(LV)感染a NSCs,用real-time PCR和Western blot实验检测shRNA对Nrf2的沉默效率。将a NSCs按照干预条件不同分为DMSO组、18α-GA组、LV-GFP组和LV-Nrf2-shRNA组,运用Brd U掺入实验、Tuj1免疫荧光染色、CCK-8实验、Hoechst 33342/PI双染色和活性氧簇(ROS)水平测定等方法评价18α-GA是否通过激活Nrf2对a NSCs的增殖、分化、细胞活力、细胞凋亡以及氧化应激水平产生影响。结果:随年龄增长,成年和老年小鼠a NSCs的Nrf2 mRNA表达水平较新生小鼠NSCs显著降低(P0.01),但a NSCs的ROS水平显著高于新生小鼠NSCs(P0.05)。应用18α-GA后,a NSCs的Nrf2 mRNA与蛋白表达水平较DMSO组明显升高(P0.01),并且a NSCs的Brd U阳性率也显著增加(P0.01)。2 mg/L 18α-GA作用后,a NSCs的Tuj1阳性率和细胞活力较DMSO组显著升高(P0.05),而凋亡率和ROS水平显著下降(P0.05和P0.01)。但是,敲减a NSCs的Nrf2并应用18α-GA干预后,LV-Nrf2-shRNA组Brd U阳性率、Tuj1阳性率以及细胞活力均较LV-GFP组显著下降(P0.05),ROS水平却显著上升(P0.05)。结论:18α-GA可能通过激活Nrf2提高a NSCs的抗氧化能力,促进a NSCs增殖和分化,维持a NSCs潜能。     

GDF15表达下调促进人胶质母细胞瘤细胞系U87MG增殖

目的探讨生长分化因子15(GDF15)表达下调对人胶质母细胞瘤U87MG细胞系增殖的影响。方法选取稳定下调GDF15的人胶质母细胞瘤U87MG细胞作为shGDF15组,以scramble细胞作为对照组。Western blot检测GDF15蛋白表达;增殖曲线和BrdU掺入实验检测细胞增殖;Western blot检测ERK1/2和p-ERK1/2蛋白表达;CCK-8实验检测细胞增殖。结果与scramble组相比较,shGDF15组细胞增殖明显加快(P0.05);细胞周期S期DNA合成增加(P0.01);ERK通路激活水平明显增加(P0.05);对化疗药物VM-26的耐受性明显增强(P0.05),并且ERK通路抑制剂可降低GDF15表达下调促进细胞增殖作用。结论GDF15可通过下调ERK通路抑制人胶质母细胞瘤U87MG细胞S期DNA合成及细胞增殖,可作为提高胶质瘤临床化疗敏感性的潜在靶点。     

FoxO1 基因对Jurkat 细胞FoxO1-KLF2-S1P1 通路的调节作用

目的:通过构建FoxO1 表达和干扰慢病毒载体,建立细胞内FoxO1-KLF2-S1P1 信号通路调控研究模型,观察FoxO1 过表达、干扰表达在Jurkat 细胞内对其下游分子表达及功能的影响。方法:构建FoxO1 表达和干扰表达慢病毒载体,分别感染Jurkat 细胞,采用荧光定量PCR、Western blot 和流式细胞术检测S1P1、CD62L、CCR7、CD69 mRNA 水平和蛋白分子的表达。结果:FoxO1 过表达组于感染后120 h FoxO1、KLF2、S1P1 和CD62L mRNA 水平显著增高(P<0.05),FoxO1、FoxO1-p 和KLF2 胞浆蛋白水平增高,S1P1+细胞和CD62L+ 细胞比率增高(P<0.05),CCR7+ 细胞和CD69+ 细胞未见显著改变(P>0.05)。FoxO1 干扰组于转染后120 h FoxO1、KLF2、S1P1 和CD62L mRNA 水平降低(P<0.05),FoxO1、FoxO1-p 和KLF2 胞浆蛋白水平低于对照组,S1P1+细胞百分比增多(P<0.05) ,但S1P1+细胞和CD62L+细胞在72 h 时减少(P<0.05)。结论:FoxO1 表达和干扰慢病毒载体转染Jurkat 细胞并调节KLF2、S1P1 和CD62L 等分子的表达,为开展细胞内FoxO1-KLF2-S1P1 信号通路调控和细胞相关功能的研究打下了基础。     

先兆子痫患者HLA-DQA1、-DQB、-DPA1基因多态性

目的：探讨人类白细胞抗原HLA-DQA1、-DQB1、-DPA1基因多态性与先兆子痫发病的关系。方法：采用序列特异性引物技术（PCR-SSP） 对46例先兆子痫患者和105例正常孕妇及其新生儿进行HLA-DQ-DPA1等位基因分型。结果：所有标本共检出11种HLA-DQA1基因表型、16种HLA-DQB1基因表型、6种HLA-DPA1基因表型。先兆子痫患者HLA-DQB1＊0301基因频率高于正常孕妇，差异有显著性（Pc=0．032，RR=2．43，AR=0．30），其余各基因表型频率两组比较差异均无显著性。结论：HLA-DQB1＊0301基因可能是一种先兆子痫发病的易感基因。     

Molecular analysis of HLA-DRB1, -DQA1 and -DQB1 polymorphism in Turkey

We report the evaluation of MHC class II polymorphism in the population of Turkey. HLA-DRB1, -DQA1 and -DQB1 have been investigated by polymerase chain reaction and sequence-specific oligonucleotide probe hybridisations (PCR/SSO) and sequence-specific priming (SSP) in 250 randomly selected healthy individuals. We also report the allelic distribution of these genes. The most frequent alleles detected were DRB1*1101 (0.104), *0301 (0.092), *0701 (0.090), DQA1*0501 (0.334), *0102 (0.164) and *03 (0.148) and DQB1*0301 (0.256), *02 (0.164), *0302 (0.128). The frequent 'putative' three-locus haplotypes carry the most frequent alleles at these loci. The most frequently detected class II "haplotypes" are DRB1*1101 DQA1*0501 DQB1*0301 (0.100), DRB1*0301 DQA1*0501 DQB1*02 (0.092) and DRB1*0701 DQA1*0201 DQB1*02 (0.072). The distribution of alleles and 'putative' haplotypes has shown common features with other Mediterranean populations. The results extend the HLA map to another Mediterranean country and provide a database for further HLA-disease association studies and transplantation applications.     

Differential regulation of CYP1A1 and CYP1B1 expression in resveratrol-treated human medulloblastoma cells

Resveratrol induces differentiation and Fas-independent apoptosis of medulloblastoma cells by a largely unknown mechanism. CYP1A1 and 1B1 are involved in resveratrol-mediated tumor suppression but their expression in medulloblastoma cells and their relevance to anti-medulloblastoma activity of resveratrol have not been described. The statuses of CYP1A1 and 1B1 in UW228-3 medulloblastoma cells without and with resveratrol treatments were elucidated in this study with ethoxyresorufin O-deethylation assay, followed by RT-PCR, immunocytochemical staining and Western blot hybridization. CYP1A1/1B1 enzymatic activity was low in UW228-3 cells but became several folds higher upon resveratrol treatments. CYP1A1 was undetectable and CYP1B1 was expressed in normally cultured cells. Accompanied by the increased fraction of apoptosis, enhanced CYP1A1 and downregulated CYP1B1 were observed in resveratrol-treated cells in time- and dose-related fashions. Our results demonstrate for the first time that in the medulloblastoma cell system, CYP1A1 upregulation is paralleled with resveratrol-induced differentiation and apoptosis, while CYP1B1 may not be an essential element in metabolic activation of resveratrol in those cells. CYP1A1 and 1B1 are resveratrol response genes and potential chemosensitive markers of medulloblastoma cells.     

Role of Niemann-Pick C1–Like 1 (NPC1L1) in Intestinal Sterol Absorption

Absorption of cholesterol by the proximal small intestine represents a major pathway for entry of cholesterol into the body pools. This cholesterol is derived primarily from the bile and diet. In adult humans, typically several hundred milligrams of cholesterol reach the liver from the intestine daily, with the potential to impact the plasma low-density lipoprotein cholesterol (LDL-C) concentration. There are three main phases involved in cholesterol absorption. The first occurs intraluminally and culminates in micellar solubilization of unesterified cholesterol, which facilitates its movement up to the brush border membrane (BBM) of the enterocyte. The second phase involves transport of cholesterol across the BBM by Niemann-Pick C1 Like-1 (NPC1L1), whereas the third phase entails a series of steps within the enterocyte involving esterification of cholesterol and its incorporation, along with other lipids and apolipoprotein B-48, into nascent chylomicrons. Discovery of the role of NPC1L1 in intestinal sterol transport occurred directly as a consequence of efforts to identify the molecular target of ezetimibe, a novel, potent, and specific inhibitor of sterol absorption that is now widely used in combination therapy with statins for management of hypercholesterolemia in the general population. Some aspects of the role of NPC1L1 in cholesterol absorption nevertheless remain controversial and are the subject of ongoing research. For example, one report suggests that NPC1L1 is located not in the plasma membrane, but intracellularly, where it is believed to be involved in cytosolic trafficking of cholesterol, whereas another concludes that a protein other than NPC1L1 is responsible for the high-affinity binding of cholesterol on intestinal BBM. However, other new studies that show that in vivo responsiveness of different species to ezetimibe correlates with NPC1L1 binding affinity further support the widely held belief that NPC1L1 does facilitate sterol uptake by the enterocyte and is the target of ezetimibe. Added to this is the unequivocal finding that deletion of the gene for NPC1L1 in mice results in near complete prevention of cholesterol absorption and an accelerated rate of fecal neutral sterol excretion. In summary, the development of ezetimibe and the identification of NPC1L1 as a key player in sterol absorption have taken research on the molecular control of this pathway to an exciting new level. From this, it is hoped that we will now be able to determine more precisely what effect, if any, other classes of lipid-lowering agents, particularly the statins, might exert on the amount of intestinal cholesterol reaching the liver.     

中国海南黎族群体HLA-DRB1、DQA1、DQB1基因多态性研究

[摘要] 目的 了解海南黎族群体HLA-DRB1、DQA1、DQB1基因的遗传多态性。方法 应用PCR-SSP方法对随机抽取的94名海南黎族无血缘关系的健康个体进行HLA-DRB1、DQA1、DQB1基因分型。结果 鉴定了海南黎族群体HLA-DRB1位点的16种等位基因,DQA1位点的10种等位基因,DQB1位点的19种等位基因,包括了DR、DQ位点目前已知的全部血清学特异性,三个位点的基因型分布均符合Hardy-Weinberg平衡（P>0.5）。结论 提供了一套比较完整准确的海南黎族群体DRB1、DQA1、DQB1等位基因的基因频率和连锁不平衡参数。对群体遗传和疾病关联研究具有参考意义。     

先兆子痫患者HLA-DQA1、-DQB1、-DPA1基因多态性

目的:探讨人类白细胞抗原HLA-DQ-A1、DQB1、DPA1基因多态性与先兆子痫发病的关系。方法:采用序列特异性引物技术(PCRSSP)对46例先兆子痫患者和105例正常孕妇及其新生儿进行HLA-DQ-DPA1等位基因分型。结果:所有标本共检出11种HLADQA1基因表型、16种HLADQB1基因表型、6种HLADPA1基因表型。先兆子痫患者HLA-DQ-B10301基因频率高于正常孕妇,差异有显著性(Pc=0.032,RR=2.43,AR=0.30),其余各基因表型频率两组比较差异均无显著性。结论:HLADQB10301基因可能是一种先兆子痫发病的易感基因。     

2009H1N1 Infection in a 1-day-old neonate

A full-term female neonate was delivered with meconium stained amniotic fluid by cesarean section by a 2009H1N1 positive 22-year-old second gravida mother, who developed symptoms 8 days prior to delivery. The neonate was completely and immediately isolated from the mother after delivery. Oseltamivir was started at birth to the neonate who had a potential possibility of 2009H1N1 infection. At 5 hours of life, the neonate developed respiratory distress. The neonate's throat swab sent for 2009H1N1 by real-time polymerase chain reaction (RT-PCR) assay was positive. The neonate required oxygen by hood for 3 days and made an uneventful recovery. The mother developed acute respiratory distress syndrome after delivery, requiring ventilatory care for 14 days and was discharged after 25 days stay in hospital. 2009H1N1 infection, although rare, needs a high index of suspicion and prompt therapy in neonates. Clinicians should be alert about the possibility of perinatal transmission of 2009H1N1.     

Latency and reactivation of bovine herpesvirus 1 (BHV-1) in goats and of caprine herpesvirus 1 (CapHV-1) in calves

Summary.  Bovine Herpesvirus 1 (BHV-1) and Caprine Herpesvirus 1 (CapHV-1) are related members of the herpesvirus family. Since their natural hosts are often kept in close contact with each other, concern was raised that a reservoir might be established in the heterologous host in addition to the homologous host. To investigate this possibility, cross-infection experiments with BHV-1 in goats and CapHV-1 in calves were performed. BHV-1 infected goats developed mild disease signs during acute infection, whereas CapHV-1 infection in calves took a subclinical course. However, virus excretion and antibody production were indicative of successful cross-infection of both BHV-1 and CapHV-1. Reactivation of BHV-1 was achieved in 5 out of 8 goats as demonstrated by recurrent virus excretion and rising antibody titers. In constrast CapHV-1 in calves could not be reactivated experimentally. Nevertheless, PCR revealed that both viruses established latency in the trigeminal ganglia of the heterologous host. Based on these results we conclude that goats should indeed be regarded as a potential BHV-1 reservoir, which must be considered during IBR eradication programs. Received July 22, 2000 Accepted January 3, 2001