病毒蛋白质类

摘要： 蛋白质泛素化广泛存在于细胞内,是蛋白翻译后的一种修饰.HBV及其相关蛋白的泛素化研究越来越受到重视.对HBV及其相关蛋白的泛素化修饰进行了综述,为进一步了解HBV的复制调控及其蛋白的泛素化研究提供了借鉴,为治愈慢性HBV感染提供新的思路和方法.

摘要： Objective To investigate the effect of interleukin-10(IL-10) on the proliferation of HL-7702 cells stably expressing hepatitis B virus X protein(HBx) gene.Methods Cell-counting Kit-8(CCK-8) assay were explored to detect the effect of HBx gene on the proliferation of HL-7702 cells and the effect of IL-10 on the proliferation of HL-7702 cells stably expressing HBx gene.Flow cytometry was used to analyze the effect of IL-10 on the apoptosis of HL-7702/HBx cells.RT-PCR was used to detect the effect of HBx gene on the expression of CDKN1B mRNA in HL-7702 cells and the effect of IL-10 on the expression of CDKN1B mRNA in HL-7702/HBx cells.Result The data of CCK-8 assay showed that the proliferation rate of HL-7702/HBx cells was significantly faster than HL-7702 and HL-7702/MOCK cells(P<0.05).The proliferation of the HL-7702/HBx cells was significantly inhibited after being treated with 80 ng/mL IL-10 for 24 hours(P<0.05).In addition,treatment of IL-10 of 80 ng/mL for 24 hours had no effects on the apoptosis of HL-7702,HL-7702/MOCK and HL-7702/HBx cells.Compared with HL-7702 and HL-7702/MOCK cells,the expression of CDKN1B mRNA level in HL-7702/HBx cells was decreased(P<0.05).After the treatment of 80 ng/mL IL-10 for 24 hours,the expression of CDKN1B mRNA level in HL-7702/HBx cells had increased(P<0.05).Conclusions HBx gene can promote the proliferation of HL-7702 cells.IL-10 can inhibit the proliferation of HL-7702/HBx cells but has no effect on the apoptosis.HBx gene probably enhances the proliferation of HL-7702/HBx cells by down-regulating the expression of CDKN1B mRNA.IL-10 can inhibit the proliferation of HL-7702/HBx cells which was probably related to the up-regulation of the expression of CDKN1B mRNA.%目的 探讨白细胞介素-10(IL-10)在稳定转染乙型肝炎病毒X(HBx)基因的HL-7702肝细胞增殖的作用及机制.方法 CCK-8法测定HBx基因对HL-7702细胞增殖的作用及IL-10对HL-7702/HBx细胞增殖的作用;流式细胞术测定IL-10对HL-7702/HBx细胞凋亡的影响;RT-PCR法测定HBx基因对HL-7702细胞表达CDKN1B蛋白质mRNA的作用及IL-10对HL-7702/HBx表达CDKN1B mRNA的作用.结果 CCK-8法显示,HL-7702/HBx细胞比HL-7702细胞和HL-7702/MOCK细胞增殖速度明显加快(P<0.05);80 ng/mL的IL-10作用24 h可抑制HL-7702/HBx细胞增殖(P<0.05);流式细胞术显示,80 ng/mL的IL-10作用24 h对HL-7702细胞、HL-7702/HBx细胞及HL7702/MOCK细胞凋亡均无影响;RT-PCR法显示,HL-7702/HBx细胞CDKN1B mRNA表达量较HL-7702细胞和HL-7702/MOCK细胞降低(P<0.05),80 ng/mL的IL-10作用24 h后,HL-7702/HBx细胞较HL-7702/HBx空白组CDKN1B mRNA表达量上调(P<0.05).结论 HBx基因可促进HL-7702细胞增殖,IL-10可抑制HL-7702/HBx细胞增殖而对其凋亡无影响.HBx基因可能通过下调HL-7702/HBx细胞CDKN1B mRNA的表达量而促进细胞增殖,IL-10可能通过上调HL-7702/HBx细胞CDKN1B mRNA的表达量而抑制细胞增殖.

摘要： 目的 探讨Wnt/β-catenin通路在稳定表达乙型肝炎病毒x蛋白(HBx)的HepG2肝癌细胞增殖中的作用机制. 方法 验证前期构建的HepG2/HBx细胞株能稳定表达HBx蛋白.CCK 8法检测HepG2/HBx,HepG2/mock及HepG2 3组细胞增殖情况,RT-PCR及Western-blot法分别检测上述3种细胞中β-cateninmRNA和蛋白表达水平;最后检测β-catenin选择性抑制剂XAV939(20 μmol/L)对各组细胞增殖水平的影响. 结果 前期构建的HepG2/HBx细胞株能稳定表达HBx蛋白.细胞增殖实验表明,HepG2/HBx组细胞增殖能力强于对照组(P＜0.05).RT-PCR及Western-blot检测结果显示,HepG2/HBx组细胞中β-catenin的mRNA及蛋白表达水平均高于对照组(P＜0.05),而HepG2/mock及HepG2组细胞之问则无明显差别.XAV939能够抑制各组细胞的增殖能力,HepG2/HBx组细胞在加入20 μmol/L XAV939作用后的增殖速度较加入相同浓度DMSO的HepG2/HBx组细胞下降(P＜0.05).XAV939对HepG2/HBx组细胞增殖的抑制强于对照组. 结论 HBx蛋白可以上调肝细胞β-catenin表达,激活Wnt邝-eatenin通路,促进肝癌细胞增殖.选择性β-catenin抑制剂可部分逆转该作用.%Objective To observe the effect of Wnt/β-catenin pathway on the proliferation of HepG2 cells expressing hepatitis B virus x protein (HBx) stably.Methods We verified that HepG2/HBx cell lines constructed previously in our lab could stably express HBx protein.Then,cell-counting Kit-8 (CCK-8) assay was used to detect the proliferation of HepG2/HBx,HepG2/mock and HepG2 cells.RT-PCR and Western-bolt were used to examine the level of β-catenin mRNA and protein expression in cells of the three cell groups.Finally,the level of cell proliferation was measured respectively after treated with 20 μmol/L β-catenin selective inhibitor XAV939 for 24 h.Results HepG2/HBx cell lines constructed previously in our lab could stably expressing HBx protein.CCK-8 assay displayed that the proliferation rate of HepG2/HBx was higher than that of HepG2/mock and HepG2 cells.The level of β-catenin mRNA and protein expression was greater in HepG2/HBx cells compared with that of HepG2/mock and HepG2 cells(P＜0.05).β-catenin selective inhibitor XAV939 suppressed cell growth of the three groups.HepG2/HBx cells with XAV939 showed lower proliferation rate than that with DMSO.The proliferation inhibition rate of HepG2/HBx cells was significantly higher than that of other two groups at three time points(P＜0.05).Conclusion The results indicate that HBx up-regulates β-catenin expression to activate wnt/β-catenin pathway and then promotes HepG2 cells proliferation.

摘要： Objective To discuss the relationship between Borna disease virus (BDV) infection and the patients of encephalitis with mental and behavioral disorders,as well as to analyze the clinical features of patients infected by BDV.Methods The p24 and p40 gene fragment of BDV in 36 patients with encephalitis,34 healthy donors and 30 patients with spinal anaesthesia were examined in peripheral blood mononuclear cells (PBMCs) and cerebrospinal fluid mononuclear cells by nested fluorescence quantitativenest reverse transcription-PCR.In the meantime,β-actin in specimens was detected by PCR as an internal reference.The clinical characteristics of patients with encephalitis with mental and behavioral disorders infected by BDV were summarized.The patients with positive mental symptoms were given a mark with the Positive And Negative Syndrome Scale.Results The positive rate (4/36,11.1％) of BDVp24 and BDVp40 in PBMCs and cerebrospinal fluid mononuclear cells in mental encephalitis was significantly higher than that in healthy donors and patients with spinal anaesthesia (0/34,0/30;P =0.015),and the number of copies of BDVp24 and BDVp40 in PBMCs was more than 102 kb/μl.One patient was positive in cerebrospinal fluid of BDVp24 gene fragment but was negative in PBMCs of BDVp40.The homologies of the gene sequence of positive product were genetically related to the virus strain BDV from horse.The clinical features of encephalitis patients with positive products were mainly hallucination and delusion.Conclusions BDV infection is correlated with encephalitis patients with mental and behavioral disorders.The clinical features of patients with BDV infection were mainly positive mental symptoms.%目的 探讨以精神行为异常为主要起病方式的脑炎患者与博尔纳病病毒(BDV)感染的相关性,分析感染BDV的脑炎患者临床特征.方法 采用荧光定量巢式逆转录聚合酶链反应方法检测36例有精神行为异常的脑炎患者(患者组)外周血和脑脊液有核细胞中BDVp24和BDVp40基因片段.以B-肌动蛋白作为内参照,同时检测34名健康体检者的外周血和30例实行腰麻患者的脑脊液(对照组)有核细胞中BDVp24和BDVp40基因片段.对BDVp24和BDVp40基因片段均为阳性的标本进行基因测序分析,外周血进行BDV抗体滴度测定,对患者的精神症状采用阳性与阴性症状量表评分,并总结阳性患者的临床特征.结果 4例以精神行为异常为主要起病方式的脑炎患者外周血和脑脊液标本BDVp24、BDVp40基因片段检测均为阳性,阳性率为11.1％ (4/36),有1例脑脊液BDVp24阳性而外周血单个核细胞中BDVp40基因片段检测阴性,拷贝数均＞102 kb/μl.对照组检测BDVp24、BDVp40基因片段均为阴性(0/34、0/30),两组阳性率比较差异有统计学意义(P=0.015).对BDVp24和BDVp40基因片段均为阳性的标本进行测序后,发现与马源的BDV病毒株亲缘关系最近.阳性标本的精神症状主要以幻觉、妄想等阳性症状为主,阳性与阴性症状量表评分均在40分以上.结论 以精神行为异常起病的脑炎患者与BDV感染有一定的相关性.BDV感染患者主要以阳性精神症状为主.

摘要： Objective To investigate the effect of cyclooxygenase‐2(COX‐2) on the cell prolifera‐tion in HL7702 cells w hich express hepatitis B virus X protein (HBx ) stably . Methods Cell‐counting Kit‐8 (CCK‐8) assay and clone formation assay were explored to detect the proliferation of HL 7702/HBx , HL7702/Mock and HL7702 cells ,RT‐PCR and Western‐bolt were used to examine the level of COX‐2 mRNA and protein expression in cells of three groups . The level of cell proliferation and COX‐2 protein expression were measured respectively after treated with selective COX‐2 inhibitor NS‐398 . Results CCK‐8 assay and plate colony formation assay displayed the proliferation rate of HL 7702/HBx was higher than that of HL7702/Mock and HL7702 cells . The level of COX‐2 mRNA and protein expression was greater in HL7702/HBx cells compared with that of HL7702/Mock and HL7702 cells(P<0 .05) . COX‐2 selective inhibitor NS‐398 suppressed growth of three groups of cells in a time‐dependent manner . The proliferation inhibition rate of HL7702/HBx cells was significantly higher than that of other two groups in three time points respectively (P<0 .05) . After treated with 50 μmol/L NS‐398 for 72 h ,COX‐2 expres‐sion was suppressed in all group , however , this phenomenon is more obvious in HL7702/HBx cells (P<0 .05) . Conclusion Regulate the expression of COX‐2 is one of the pathway for the effect of HBx on the HL7702 proliferation ,COX‐2 inhibitor can suppress the cell proliferation for the greater part in‐duced by this pathway .%目的：探讨环氧合酶‐2（COX‐2）在稳定表达乙型肝炎病毒X蛋白（HBx）的HL7702肝细胞增殖中的作用及机制。方法 CCK‐8法及克隆形成实验检测HL7702／HBx细胞、HL7702／Mock细胞、HL7702细胞增殖情况，RT‐PCR法检测上述3种细胞中 COX‐2 mRNA 表达水平，Western‐blot 法检测 COX‐2蛋白表达水平， COX‐2选择性抑制剂NS‐398作用于各组细胞后再检测以上各组细胞增殖情况及COX‐2蛋白表达水平的改变。结果细胞增殖实验及克隆形成实验提示，HL7702／HBx 组细胞增殖能力强于 HL7702／Mock和 HL7702组（P＜0．05），克隆形成率也更高（P＜0．05）。RT‐PCR检测结果提示，相对于 HL7702／Mock细胞和 HL7702组细胞，HL7702／HBx组细胞内COX‐2的mRNA相对表达量明显增高（P＜0．05）。Western‐blot检测提示，HL7702／HBx组细胞中的COX‐2蛋白相对表达水平较高（P＜0．05），而 HL7702／Mock及 HL7702组细胞之间则无明显差别。NS‐398以时间依赖的方式部分抑制各组细胞的增殖能力，对 HL7702／HBx组细胞增殖的抑制强于其他2组细胞（P＜0．05）。经50μmol／L NS‐398处理后，3组细胞的COX‐2蛋白水平均显著降低，此现象在 HL7702／HBx组细胞中更加明显（P＜0．05）。结论 HBx蛋白可以上调肝细胞COX‐2表达，促进肝细胞增殖，选择性COX‐2抑制剂可部分逆转该作用。

摘要： 宫颈癌是妇科肿瘤中的第二大常见肿瘤,现已明确人乳头瘤病毒(HPV)感染是宫颈癌的主要致病因素,但并非所有HPV感染都会导致官颈癌.HPV L1蛋白是HPV的主要衣壳蛋白,其表达与HPV的复制及其早期感染密切相关.近年研究表明,在目前细胞学基础上检测病变组织HPV L1蛋白可提高官颈病变筛查及预测病变进展的准确度,预防官颈病变的过度诊治.综述HPV L1在预测官颈病变的发展及预后的科学性及实用性,以寻找更加合理的筛查流程,对不同患者选择不同的治疗方案.%Cervical cancer is the second most common tumor in gynecologic oncology,HPV L1 protein is the major capsid protein of HPV,and it's expression is closely related to viral replication and early infection. Recent study shows that present screening method simultaneously unites HPV L1 capsid protein detection may enhances the accuracy for the screening cervical disease and prediction of progression of lesions. This review is concentrated on update scientific nature and the usability of the research for detection of HPV L1 capsid protein in predicting the prognosis and development of cervical disease a review, looking for the more reasonable screening flow, and choosing the different therapeutic schedule for different patients.

摘要： Objective To investigate the effects of hepatitis B virus X protein (HBx) on the pro-liferation of human hepatic stellate cells (HSCs) and to discuss possible mechanism (s) in the pathway . Methods The eukaryotic expression vector of HBV X gene (p HBV-X-IRES2-EGFP) was transfected in-to human liver cell HL-7702 and selected by G418 to establish a cell line L02/x ,which was confirmed to stably express H BV X gene by RT-PCR and Western blot analysis . HSCs were co-cultured with L02/x cells ,empty vector-transfected cells and non-transfected cells for 36 h ,and were named HSC/x ,HSC/ctr and HSC/NC respectively . Then the proliferation of HSCs in these three groups were detected by CCK8 method and mRNA expressions of TGFβ1 and CTGF in HSCs were analyzed by Real-time PCR . Results RT-PCR and Western blot analysis showed L02/x cells expressed HBV X gene steadily . The prolifera-tions of HSC/x (0 .737 7 ± 0 .014 2)were significantly higher than those of HSC/ctr(0 .497 0 ± 0 .008 5) and HSC/NC(0 .491 3 ± 0 .018 6) ,P<0 .01 . Real-time PCR showed mRNA levels of TGFβ1 and CTGF in HSC/x (3 .146 1 ± 0 .070 5 ,2 .982 9 ± 0 .031 5) were significantly higher than those in HSC/ctr(1 .004 6 ± 0 .004 0 ,1 .022 2 ± 0 .062 7)and HSC/NC (1 .000 0 ± 0 .000 0 ,1 .000 0 ± 0 .000 0) ,both P< 0 .01 . Conclusion The expression of HBV X gene in HL-7702 cells can promote the proliferation of co-cultured HSCs through up-regulating mRNA expressions of TG Fβ1 and CTG F .%目的研究乙肝病毒X蛋白（HBx）对肝星状细胞（HSC）增殖的影响及其可能的分子机制。方法运用分子生物学方法构建稳定表达 H BV X 基因的 HL-7702肝细胞株（L02/x ）。RT-PCR、Western blot 鉴定L02/x细胞 H BV X基因的稳定表达。将HSC细胞分别与L02/x、转染空质粒和未转染的肝细胞共培养36 h ，分别命名为HSC/x、HSC/ctr和HSC/NC。CCK-8法检测各组HSC增殖，Real-time PCR法检测在HSC增殖中起重要作用的 TGFβ1和 CTGF基因在各组HSC中的mRNA表达。结果 RT-PCR和Western blot结果显示，L02/x细胞中有 HBV X基因mRNA和蛋白的表达。CCK-8法检测显示，HSC/x的增殖（0．7377±0．0142）显著高于HSC/ctr（0．4970±0．0085）和HSC/NC（0．4913±0．0186），差别具有统计学意义（P＜0．01）。Real-time PCR显示，HSC/x细胞中的 TGFβ1和 CTGF mRNA相对表达量（3．1461±0．0705，2．9829±0．0315）均显著高于HSC/ctr（1．0046±0．0040，1．0222±0．0627）和HSC/NC（1．0000±0．0000，1．0000±0．0000），差别具有统计学意义（P均＜0．01）。结论肝细胞中表达的 HBV X基因可以上调 HSC细胞 TGFβ1和 CTGF基因的表达，促进共培养的 HSC增殖。

摘要： Objective To construct and express the recombinant encoding C protein derived from Japanese encephalitis virus (JEV). Methods Gene encoding C was amplified by RT-PCR technique from JEV SA14-14-2 strain total RNA. JEV C protein gene was obtained with restriction cndonuclcasc BamH I/EcoR I from the pro-karyotic expression vector named after Pmd19-T simple, and subcioncd into pcDNA3. 1( + ) cukaryotic vector, named after Pjc. The recombinant was confirmed by restriction enzymes analysis and DNA sequencing, then was transfected into China hamster ovary (CHO) cells by Lipofcctaminc 2000. Distribution and expression of the C protein encoded by Pjc in transfced CHO cells were detected by immunofluorcsccncc. Results DNA fragments (414bps)of the inserts released from Pjc with BamHI/EcoRI restriction cndonuclcasc were the expected theoretic results. The expression of above C protein was mainly distributCd in cndochylcma of transfected CHO cells, and not much in membrane of transfected CHO cells. Conclusion The recombinant Pjc was constructed and transfected into CHO cells successfully, and transfected CHO cells can express JEV C protein.%目的 构建流行性乙型脑炎病毒(JEV)C蛋白编码基因重组子并鉴定.方法 以JEV SA14-14-2株总RNA为模板,应用反转录-聚合酶链反应(RT-PCR)方法扩增JEV C蛋白编码基因,克隆至pMD19-T Simple载体测序.为便于分析JEV C蛋白编码基因重组子在哺乳动物细胞中的表达,在JEV C蛋白编码基因5′端附加FLAG 序列,并亚克隆至pcDNA 3.1(+)载体中,构建重组子pJc并经酶切及DNA测序分析.脂质体法将pJc转染中华仓鼠卵巢(CHO)细胞.免疫荧光检测转染的CHO细胞中JEV C蛋白分布与表达.结果 pJC经BamH I/EcoR I酶切释出的插入子片段(414 bp)分别与预期结果相符合.所编码的融合蛋白主要分布于胞质,少量分布于胞膜.结论 pJC成功构建,转染的CHO细胞可表达JEV C蛋白.

摘要： Objective To determine the pathogen of a local dengue fever outbreak in Shenzhen city in 2010,and to analyze the molecular characteristics of the epidemic dengue virus strain as well as explore the possible origin.Methods The serum samples collected from the suspect dengue fever cases were detected for IgM, IgG by enzyme-linked immunosorbent assay ( ELISA ),immunochromatography and dengue virus nucleic acid by real-time polymerase chain reaction (PCR).Serum samples from patients with early stage dengue fever were used to isolate virus with C6/36 and BHK-21 cell lines.The type of isolated virus strain was determined by RT-semi-nested-PCR and realtime PCR.E gene of isolated virus strain was amplified by RT-PCR and sequenced.Homology and phylogenetic tree of E gene of Shenzhen dengue virus with the strains isolated from other areas were constructed.Results IgM,IgG and RNA of type 1 dengue virus were detected in serum samples from dengue fever suspected patients.Type 1 dengue virus named DEV1-SZ1029 was successfully isolated from the serum sample.The homology of nucleotide sequence of E gene of SZ1029 strain with standard type 1 dengue virus HAWAII 45,Fj231/04,GD14/97 and GD05/99 were 94.8％,99.6％,97.7％ and 98.5 ％,respectively.The phylogenetic tree indicated that SZ1029 had the greatest similarity with the D1/Malaysia/36000/05 strain,SG(EHI)DED142808 strain and Fj231/04 strain and they lied in the same branch of the phylogenetic tree.The isolated dengue virus type 1 belonged to genetype Ⅰ with GZ/80,Taiwan87,All patients lived in a certain construction site in Shenzhen and had no recent travel history outside the area in one month before infection.Conclusions The virological,serological and molecular features all identify that the local dengue fever outbreak in Shenzhen in 2010 is caused by type 1 dengue virus and SZ1029 strain may be transferred from Southeast Asian region,and there may be a plague focus in Shenzhen.%目的 分析深圳市2010年登革热暴发疫情的病因,从分子水平探讨流行毒株的生物学特征,追踪其地域来源.方法 采用ELISA、胶体金免疫层析法和荧光PCR检测疑似登革热患者血清中的特异性IgM、IgG抗体和病毒核酸,并用C6/36和BHK-21细胞对早期病例血清进行病毒分离,采用反转录-半套式PCR和荧光PCR方法对其进行型别鉴定.同时扩增病毒E基因后进行序列测定,并与不同国家和地区的登革病毒株进行同源性比较和进化树分析.结果 从疑似登革热患者血清中检测到登革病毒IgM、IgG抗体及登革1型病毒核酸.深圳市登革1型病毒分离株SZ1029与登革1型国际标准株HAWAII 45株、我国福建省Fj231/04株及广东省1997、1999年登革1型病毒流行株GD14/97、GD05/99在E基因上的核苷酸同源性分别为94.8％、99.6％、97.7％和98.5％.基因进化树显示,深圳市登革病毒分离株与马来西亚分离株D1/Malaysia/36000/05、新加坡分离株SG(EHI)DEDI42808和Fj231/04株亲缘关系最近,在进化树的同一分支上,与GZ/80、Taiwan87同属基因Ⅰ亚型.所有患者发病前1个月在深圳市某工地居住,无输血史、无外出史.结论 该次疫情的病因为登革1型病毒感染,该毒株有可能来源于东南亚一带,推测深圳可能存在登革1型病毒的疫源地.

摘要： 本发明提供了包含一或多种同种型红细胞生成素(EPO)的组合物，所述红细胞生成素包含与其连接的聚糖，特征在于所述聚糖包含LewisX结构及每个EPO分子平均至少6个唾液酸部分。本发明进一步提供了获得包含一或多种同种型红细胞生成素(EPO)的组合物的方法，所述红细胞生成素包含与其连接的聚糖，其中所述聚糖包含平均每个EPO分子至少6个唾液酸及0-2个Lewis x结构，所述方法包括：a)提供含有可表达形式的编码腺病毒E1A蛋白的核酸序列及进一步含有可表达形式的编码EPO的核酸的真核细胞，其中所述细胞进一步含有在异源启动子控制下的编码唾液酸转移酶、优选α-2，6-唾液酸转移酶或α-2，3-唾液酸转移酶的核酸序列；b)在无血清培养基中培养所述细胞，使得EPO在所述细胞中表达；c)从所述细胞和/或培养基中收获表达的EPO；及d)纯化并分级分离EPO以获得每个EPO分子N联聚糖的平均唾液酸含量增加的级分，获得包含一或多种同种型EPO的组合物，所述EPO包含与其连接的聚糖，其中所述聚糖包含每个EPO分子平均至少6个唾液酸及0-2个Lewis x结构。

摘要： 本发明提供了包含一或多种同种型红细胞生成素(EPO)的组合物，所述红细胞生成素包含与其连接的聚糖，特征在于所述聚糖包含LewisX结构及每个EPO分子平均至少6个唾液酸部分。本发明进一步提供了获得包含一或多种同种型红细胞生成素(EPO)的组合物的方法，所述红细胞生成素包含与其连接的聚糖，其中所述聚糖包含平均每个EPO分子至少6个唾液酸及0—2个Lewis x结构，所述方法包括：a)提供含有可表达形式的编码腺病毒ElA蛋白的核酸序列及进一步含有可表达形式的编码EPO的核酸的真核细胞，其中所述细胞进一步含有在异源启动子控制下的编码唾液酸转移酶、优选α-2，6-唾液酸转移酶或α-2，3-唾液酸转移酶的核酸序列；b)在无血清培养基中培养所述细胞，使得EPO在所述细胞中表达；c)从所述细胞和/或培养基中收获表达的EPO；及d)纯化并分级分离EPO以获得每个EPO分子N联聚糖的平均唾液酸含量增加的级分，获得包含一或多种同种型EPO的组合物，所述EPO包含与其连接的聚糖，其中所述聚糖包含每个EPO分子平均至少6个唾液酸及0—2个Lewis x结构。

摘要： 由下述氟树脂形成了与蛋白质或包含蛋白质的组合物接触的表面的容器及用于制造蛋白质或包含蛋白质的组合物的器材具有显著的低蛋白质吸附性，其中，所述氟树脂为选自四氟乙烯‑六氟丙烯系共聚物及四氟乙烯‑全氟烷基乙烯基醚系共聚物中的至少1种氟树脂，并且，所述氟树脂的熔点为320°C以下，所述氟树脂中的相对于每1×10

摘要： 本发明提供能减小使用了特异性地结合的物质的测定中的由肝型脂肪酸结合蛋白质引起的测定值的变动幅度的肝型脂肪酸结合蛋白质制剂、评价该制剂的方法、制作肝型脂肪酸结合蛋白质的校准曲线的方法、以及定量该蛋白质的方法。一种用使用了用10mM的氧化剂在25°C下进行1小时的氧化处理的肝型脂肪酸结合蛋白质制剂的测定值与使用了未进行上述氧化处理的肝型脂肪酸结合蛋白质制剂的测定值的比表示的氧化变动系数设定为1.4以下的肝型脂肪酸结合蛋白质制剂、用于该制剂的肝型脂肪酸结合蛋白质、编码该蛋白质的DNA、由该DNA转化得到的细胞、该蛋白质的制造方法、评价该制剂的方法、抑制使用该制剂的测定中的测定值的变动幅度的方法、制作该蛋白质的校准曲线的方法、以及定量该蛋白质的方法。

摘要： 本发明提供一类能够携带生物活性蛋白分子穿透细胞膜的非肽类多胍有机小分子或其药学上可接受的盐，所述非肽类多胍有机小分子具有式I所示的结构：

摘要： 本发明提供一种糖化蛋白质测定试剂，其至少包含Trinder’s试剂、4‑氨基安替比林、蛋白酶、蛋白酶的稳定剂以及亚铁氰化物，其中，至少Trinder’s试剂包含在含有Trinder’s试剂的部分组合物中，至少4‑氨基安替比林包含在含有4‑氨基安替比林的部分组合物中，蛋白酶的稳定剂是使亚铁氰化物与该蛋白酶的稳定剂混合后的氧化还原电位大于0.058V的稳定剂，氧化还原电位是包含蛋白酶的稳定剂和亚铁氰化物、不包含糖化蛋白质的反应体系的氧化还原电位。

摘要： 本发明提供了包含一或多种同种型红细胞生成素(EPO)的组合物，所述红细胞生成素包含与其连接的聚糖，特征在于所述聚糖包含LewisX结构及每个EPO分子平均至少6个唾液酸部分。本发明进一步提供了获得包含一或多种同种型红细胞生成素(EPO)的组合物的方法，所述红细胞生成素包含与其连接的聚糖，其中所述聚糖包含平均每个EPO分子至少6个唾液酸及0-2个Lewis x结构，所述方法包括：a)提供含有可表达形式的编码腺病毒E1A蛋白的核酸序列及进一步含有可表达形式的编码EPO的核酸的真核细胞，其中所述细胞进一步含有在异源启动子控制下的编码唾液酸转移酶、优选α-2，6-唾液酸转移酶或α-2，3-唾液酸转移酶的核酸序列；b)在无血清培养基中培养所述细胞，使得EPO在所述细胞中表达；c)从所述细胞和/或培养基中收获表达的EPO；及d)纯化并分级分离EPO以获得每个EPO分子N联聚糖的平均唾液酸含量增加的级分，获得包含一或多种同种型EPO的组合物，所述EPO包含与其连接的聚糖，其中所述聚糖包含每个EPO分子平均至少6个唾液酸及0-2个Lewis x结构。

摘要： 本发明提供了包含一或多种同种型红细胞生成素(EPO)的组合物，所述红细胞生成素包含与其连接的聚糖，特征在于所述聚糖包含LewisX结构及每个EPO分子平均至少6个唾液酸部分。本发明进一步提供了获得包含一或多种同种型红细胞生成素(EPO)的组合物的方法，所述红细胞生成素包含与其连接的聚糖，其中所述聚糖包含平均每个EPO分子至少6个唾液酸及0－2个Lewisx结构，所述方法包括：a)提供含有可表达形式的编码腺病毒E1A蛋白的核酸序列及进一步含有可表达形式的编码EPO的核酸的真核细胞，其中所述细胞进一步含有在异源启动子控制下的编码唾液酸转移酶、优选α－2，6－唾液酸转移酶或α－2，3－唾液酸转移酶的核酸序列；b)在无血清培养基中培养所述细胞，使得EPO在所述细胞中表达；c)从所述细胞和/或培养基中收获表达的EPO；及d)纯化并分级分离EPO以获得每个EPO分子N联聚糖的平均唾液酸含量增加的级分，获得包含一或多种同种型EPO的组合物，所述EPO包含与其连接的聚糖，其中所述聚糖包含每个EPO分子平均至少6个唾液酸及0－2个Lewisx结构。

摘要： 本发明提供能够以优异的安全性将蛋白质导入到细胞中的蛋白质导入方法。通过使目标蛋白质承载在由粘土矿物构成的蛋白质导入用载体上并将其添加到细胞中，可以将所述目标蛋白质导入到所述细胞内中。所述粘土矿物优选为层状粘土矿物，可以使用蒙脱石、蛭石、伊利石等。

摘要： 本发明涉及与粉碎或切断洋葱等植物时发生催泪成分的生成有关的表现出将1－丙烯次磺酸转变为催泪成分的作用的蛋白质或多肽、编码该蛋白质或多肽的DNA(催泪成分合成酶基因)、使用用于属于葱属植物的催泪成分合成酶基因分离的引物以及使用该引物分离该基因的方法、含有上述DNA的重组载体、包含含有上述DNA的重组载体的转化体、对用含有上述DNA的重组载体转化的宿主细胞进行培养制造具有催泪成分合成酶活性的蛋白质或多肽的方法、具有催泪成分合成酶活性的蛋白质或多肽、以及具有抑制具有催泪成分合成酶活性的蛋白质或多肽的mRNA翻译的功能的核酸分子。