Yeast Extract from Express Five Serum-free Medium contains Factors at about 35 kDa,Essential for Growth of <Emphasis Type="Italic">Trichoplusia ni</Emphasis> Insect Cells

The yeast extract (of unknown origin) present in the commercially available serum-free medium ‘Express Five’ contains factors (‘yeast extract factors’) up to 35 kDa which are essential for growth of Trichoplusia ni insect cells. A yeast extract brand lacking these components could not support growth of T. ni cells. However, cell proliferation was restored by adding chromatographic fractions containing the yeast extract factors. The yeast extract factors were not solely responsible for the growth enhancing effect of yeast extract but some other components, which seem to be generally present in yeast extracts, are also required for T. ni proliferation.     

植物异黄酮合酶研究进展

植物异黄酮是一类具有增强植物抗病、诱导根瘤形成以及预防激素相关肿瘤发生、缓解女性更年期综合症的活性次生代谢产物,其合成关键酶是异黄酮合酶(IFS)。介绍了IFS的催化机理、基因克隆与表达调控的研究进展,讨论了开展IFS代谢工程研究对提高植物抗病虫害能力和改善农作物营养保健功效的意义。     

FOXP3 在食管癌中表达规律研究

目的:研究FOXP3在食管癌中表达规律。方法:选择2010年1月~2010年10月在我院接受手术治疗的食管鳞癌患者48例,选取所有患者肿瘤组织及正常食管黏膜组织进行免疫组化分析,观察FOXP3在食管癌细胞系Eca-109、正常食管黏膜细胞系Eca-109、人食管癌组织、正常食管黏膜组织的表达,FOXP3在不同年龄、性别、肿瘤分化程度、肿瘤直径、淋巴结转移上的表达。结果:FOXP3在食管癌组织中阳性表达率(85.42%)显著高于正常食管黏膜组织(14.58%),P0.05。FOXP3在正常细胞系表达为阴性,在食管癌细胞系Eca-109中的表达阳性,FOXP3在正常食管黏膜组织中表达阴性,在食管癌组织中表达阳性。在食管癌组织中,FOXP3阳性表达率和食管癌患者的性别、年龄等不同临床病理特征均无关联(P0.05),和T分期、TNM分期、肿瘤直径、淋巴结转移、远处转移等存在关联(P0.05)。结论:食管癌细胞能够表达FOXP3,且其表达水平和食管癌的进展关系密切,可以作为临床诊断及治疗食管癌的重要参考指标。     

TIMP-2在毕赤酵母中的克隆与表达

为了克隆人基质金属蛋白酶组织抑制剂-2（TIMP-2）基因,并在Pichia pastoris中表达,根据GenBank上的TIMP-2的氨基酸序列和毕赤酵母偏爱密码子,通过化学合成和PCR相结合的方法获得了人的TIMP-2基因全长序列,构建了pPIC9-T2表达载体,电击转化到毕赤酵母,通过表型筛选和诱导表达得到蛋白表达工程菌,并对表达产物进行了分离纯化和生物学活性分析。     

人胰岛素原在甲醇酵母(Pichia pastoris)中的高效表达

甲醇营养型酵母Ｐｉｃｈｉａ ｐａｓｔｏｒｉｓ在近十几年已被人们广泛用作外源基因表达的系统。表达的是可溶性蛋白,且胞外分泌。本研究系将外源基因（胰岛素原基因）连接到穿梭质粒ＰＨＩＬ－Ｓ１上,再通过同源重组到酵母染色体,筛选表达株。用１ 升发酵罐在甲醇诱导下可获得０．３ｇ／Ｌ胰岛素原的产率。     

神经细胞黏附因子在子宫腺肌病病灶中的表达及意义

目的:探究神经细胞黏附因子(NCAM)在子宫腺肌病病灶中的表达及意义。方法:将2016年6月~2017年6月在我院接受治疗的80例子宫腺肌病患者作为研究对象,包括分泌期与增生期各40例,采用免疫组化法检测,其在位内膜、异位内膜组织中NCAM表达情况,并以同期我院40例正常者子宫内膜标本作为对照。对子宫腺肌病患者痛经程度给予NRS疼痛评估,比较其NCAM的表达。结果:80例子宫腺肌病在位内膜、异位内膜均存在NCAM表达,40例正常内膜腺上皮有38例存在NCAM表达,2例正常内膜无表达。NCAM在异位内膜组织中的表达明显高于在位内膜及正常子宫内膜(P0.05),差异均有统计学意义。NCAM在在位内膜组织分泌期表达与增生期差异有统计学意义(P0.05);子宫腺肌病异位病灶NCAM表达与患者NRS评分呈现明显正相关(r=0.824,P0.05)。结论:NCAM在异位子宫内膜高表达,可能参与了子宫内膜异位症的发生和发展,并与患者痛经程度呈现出正相关。     

HGV RNA基因组感染HepG2细胞的研究

为了观察HGV RNA基因组在HepG2细胞中的复制和表达并建立HGV感染的细胞模型,体外转录制备HGV RNA基因组,Lipfectamin介导转染HepG2细胞。取HGV RNA阳性培养上清液传代感染HepG2细胞,采用RT-PCR、免疫组化和Western blot等技术检测HGV在HepG2细胞中的复制和表达。HepG2细胞地转染后24h便可在培养上清液中检测到HGV负链RNA,传代感染的细胞及培养上清液中可检测到HGV正、负链RNA。在90d内传代20余次,均能检测到HGV的复制。免疫组化和Western blot可检测到 HGV E2蛋白在感染细胞中的表达。HGV感染细胞经冻存后复苏,仍能检测到HGV RNA。故HGV RNA基因组能够在HepG2细胞中复制和表达,此细胞模型有可能用于HGV的复制与感染防治的研究。     

Cellular signaling in sinecatechins-treated external genital and perianal warts: unraveling the mechanism of action of a botanical therapy

<正>Dear Editor,Derived from green tea leaves,sinecatechins(Veregen)ointment,15%is a topical therapy that is FDA-approved to treat human papillomavirus(HPV)-induced external genital and perianal warts(EGW)in immunocompetent patients aged 18 years and older.In two phase 3 trials enrolling over 1,000 participants with EGW,a 16-week treatment regimen with sinecatechins ointment resulted in higher rates of complete clearance of all warts—both     

Evaluation of a recombinant excretory secretory Haemonchus contortus protein for use in a diagnostic enzyme-linked immunosorbent assay

The nematode Haemonchus contortus (H. contortus) is one of the most pathogenic and economically important parasites of sheep. A 24 kDa protein is one of the important components in H. contortus excretory/secretory (ES), which was shown to have important biological function. In our research, the cDNA of its open reading frame (ES24) was obtained and analyzed. Then the ES24 was sub-cloned into pET-30a expression vector. The recombinant vector that codes hexahistidyl peptide fusion protein (His-ES24) was transformed into Escherichia coli BL21 (DE3) strain. After induction, a high expression level of His-ES24 was found at 6h taking about 26% of the total bacterial protein analyzed by gel thin-layer scanning. The expressed His-ES24 was purified and then used in an enzyme-linked immunosorbent assay (ELISA) to detect specific antibodies in serum samples. The ELISA was able to differentiate between H. contortus-infected sheep serum and Fasciola hepatica-infected sheep serum or non-infected sheep serum. No cross-reaction was observed in sheep sera that have been experimentally infected with F. hepatica. A total of 153 field sheep serum samples conserved in our laboratory were examined using the His-ES24 ELISA, and 82 (53.6%) of them were found seropositive to H. contortus. Our results demonstrate that the prokaryotic-expressed His-ES24 might be a useful diagnostic reagent for epidemiological studies of H. contortus in sheep.     

禽流感病毒HA部分基因的克隆及其表达

禽流感病毒(Avian influenza virus,AIV)是正粘病毒科流感病毒属的成员。AIV核酸为单链线状分段RNA,病毒表面囊膜上镶嵌着两种重要纤突,即血凝素(HA)和神经氨酸酶(NA)。HA和NA与病毒的毒力、传染性密切相关。其中ha基因是最重要的免疫原性基因,它刺激机体所产生的抗体可中和病毒、抵抗感染,