以痘苗病毒天坛株为载体的表达单纯疱疹病毒Ⅰ型糖蛋白D的重组活疫苗的建立

从我国分离到的一株单纯疱疹病毒Ⅰ型（ＨＳＶ－１－１６８株）病毒基因组中,分离出含有糖蛋白Ｄ（ｇＤ）基因的１．２ｋｂ片段,插入带有痘苗病毒天坛株ＴＫ区的质粒ｐＪＳＢ１１７５Ｐ７．５ｋ启动子下游,转染无白血病鸡胚原代细胞,获得带有ＨＳＶ－１－１６８ｇＤ基因的重组痘苗病毒。此株重组病毒在感染细胞膜上表达ＨＳＶ－１－１６８ｇＤ糖蛋白抗原,能与特异性单克隆抗体反应。在感染细胞中表达的膜抗原经ＳＤＳ－ＰＡＧＥ分析,表达分子量为５４ｋＤ糖蛋白。用Ｓｏｕｔｈｅｒｎ杂交分析了重组病毒ＤＮＡ中特异的ｇＤ基因,对作为活疫苗的重组痘苗病毒株进行了一些微生物学活性、免疫原性和毒力等方面的研究。     

中国脊髓灰质炎病毒疫苗株基因变异的分析

从急性弛缓性麻痹（ＡＦＰ）病例分离的３２株脊髓灰质炎（脊灰）病毒,在ＶＰ１编码区,经ＰＣＲ－ＲＦＬＰ方法鉴定,并经核酸测序进一步证实力疫苗株,其中５株为Ｉ型疫苗,１６株Ⅱ型疫苗株,１１株Ⅲ型疫苗株,以同样的分析方法检测这些毒株基因组的３Ｄ聚合酶编码区,发现２株Ｉ型疫苗株在３Ｄ区的４３１个核苷酸序列为野毒序列,即这２株Ｓａｂｉｎ１基因型（ＶＰ１）与野毒基因型（３Ｄ）重组株;其余３株Ｉ型疫苗株未发现基     

以痘苗病毒天坛株为载体的表达单纯疱疹病毒I型糖蛋白D的重组活疫苗…

从我国分离到的一株单纯疱疹病毒Ｉ型病毒基因组中,分离出含有糖蛋白Ｄ基因的１．２ｋｂ片估,插入带有痘苗病毒天坛株ＴＫ区的持粒ｐＪＳＢ１１７５Ｐ７．５ｋ启动子下游,转染无白血病鸡胚原供细胞,获得带有ＨＳＶ－１－１６８ｇＤ基因的重组痘苗病毒。此株重组病毒在感染细胞膜上表达ＨＳＶ－１－１６８ｇＤ糖蛋白抗原,能与特异性单克隆抗体反应。     

中国脊髓灰质炎Ⅱ型疫苗相关分离株病毒性状的观察

对１９９４年中国分离的１３株脊髓灰质炎Ⅱ型疫苗相关株进行了ＰＣＲ－ＲＦＬＰ分析,发现７株为重组病毒,毒力较疫苗株有回复,在Ⅱ型脊髓灰质炎病毒基因序列上,对于神经毒力有重要影响的第４８１位核苷酸发生突变,另一个被视为重要位点的２９０８位核苷酸无一发生变化,反而在２９０９位核苷发生了高频率的点突变,意味着２９０９位点在中国Ⅱ型疫苗相关株的自然变异中可能起着重要作用。     

口服狂犬病疫苗

口服狂犬病疫苗根据毒种类型分为二类：一类为减毒活疫苗，首先美国用ＥＲＡ株在实验室研究成功，随后各国相继研究ＳＡＤ株的突变株ＳＡＤＢｅｒｎ（或ＳＡＤｓｗｉｓｓ），ＳＡＤＢ＿１９、ＳＡＧ＿１以及ＣＴＮ—１株口服狂犬病疫苗，均获得较好的口服免疫原性，对动物ＳＡＤＢｅｒｎ、ＳＡＤＢ＿１９、ＳＡＤｓｗｉｓｓ、ＣＴＮ—１的毒性相近，而ＳＡＧ＿１的毒性最弱，对多种成年野生动物不致病。以上各株病毒分别被制成口服疫苗在世界许多地区得到试验和应用。另一类为重组基因工程口服疫苗，已将狂犬病毒的糖蛋白基因分别重组到人腺病毒５型，痘苗病毒、浣熊痘病毒、金丝雀痘病毒、多角体病毒中，并证实均有免疫原性，重组痘病毒ＶＶＴＧｇＲＡＢ制备的口服疫苗已在比利时进行大规模的区域性试验，具有有效性、无毒性和热稳定性。口服疫苗的诱饵多采用鸡头、蜡制或膨化食物外壳内包装有疫苗胶囊的复合体。     

我国单纯疱疹病毒Ⅰ型168株糖蛋白D基因的克隆及其在痘苗病毒天坛株中的表达

将我国单纯疮疹病毒Ⅰ型１６８株基因组ＤＮＡ中扩增出的糖蛋白Ｄ基因,插入痘苗病毒ｐ７．５启动子下游,使其在痘苗病毒天坛株表达。免疫荧光分析表明,产生的重组病毒糖蛋白Ｄ能被运到被重组病毒感染的１４３细胞表面表达,表达的产物经Ｗｅｓｔｅｍｂｌｏｔ鉴定为分子量约５０ｋＤ的多肽。Ｓｏｕｔｈｅｍｂｌｏｔ证明重组病毒基因组中整合有ＨＳＶ－１１６８株糖蛋白基因片段,重组病毒免疫家兔后,产生了高滴度的ＨＳＶ特异性中和抗体。     

我国单纯疱疹病毒I型168株糖蛋白D基因的克隆及其在痘苗病毒天坛…

将我国单纯疱疹病毒Ｉ型１６８株基因组ＤＮＡ中扩增出的糖蛋白Ｄ基因,插入痘苗病毒ｐ７．５启动子下游,使其在痘苗病毒天坛株表达,免疫荧光分析表明,产生的重组病毒糖蛋白Ｄ能被运到被重且病毒感染的１４３细胞表面表达,表达的产物经Ｗｅｓｔｅｎｂｌｏｔ鉴定为分子量约５０ｋＤ的多肽,Ｓｏｕｔｈｅｒｎｂｌｏｔ证明重组病毒基因组中整合有ＨＳＶ－１１６８株糖蛋白基因片段,重组病毒免疫家兔后,产生了高滴度的ＨＳＶ特异性     

HIV－1外膜（env）基因在重组痘苗病毒中的高效表达

应用重组痘苗病毒作载体,在ＡＴＩ、Ｐ７．５突变型早期启动子控制下,高效表达了（经定点突变去除一处早期终止信号的）ＨＩＶ－１ｅｎｖ基因。经Ｗｅｓｔｅｒｎｂｌｏｔ证明,晚期启动子活性强的ｐＳＦＪ２－３８能更高效地表达ｅｎｖ基因。经Ｌｅｎｔｉｌ－Ｌｅｃｔｉｎ提取以及ＳＤＳ－ＰＡＧＥ后的激光扫描测定结果,在１０７个感染细胞中可产生５０－７０μｇ的Ｅｎｖ蛋白。     

球形幽门螺杆菌分子生物学研究

为研究幽门螺杆菌（ＨＰ）球形变异本质,作者通过延期培养和采用亚抑菌浓度抗生素,使３株ＨＰ发生球形变异,对弯曲形和球形ＨＰ作了ＳＤＳ－ＰＡＧＥ、免疫印迹及４个毒力基因片段ＰＣＲ和ＰＣＲ－ＳＳＣＰ分析。ＳＤＳ－ＰＡＧＥ图谱显示球形ＨＰ分子量在７４×１０４以上的蛋白含量减少,免疫印迹显示球形ＨＰ１２５×１０４蛋白条带反应减弱,而抗生素诱变的球形ＨＰ分子量为１１×１０４和６３×１０４的蛋白条带反应增强。ＰＣＲ及ＰＣＲ－ＳＳＣＰ结果表明球形ＨＰ的ｈｐａＡ,ＶａｃＡ,ＣａｇＡ和ＵｒｅＡ４个毒力基因片段未发生缺失,但在ｈｐａＡ或ＶａｃＡ基因中存在点突变     

表达HSV－2 gD基因的重组痘苗病毒保护小鼠对抗致死量HSV病霉攻击

利用ＰＣＲ方法对单纯疱疹病毒Ⅱ型糖蛋白Ｄ（ＨＳＶ－２ｇＤ）基因进行了修饰,在其５＇端删去约５００ｂｐ的非编码区,仅保留ＡＴＧ上游７个ｂｐ。将修饰后的ＨＳＶ－２ｇＤ基因插入到带有痘苗病毒天坛株ＴＫ基因区段的痘苗表达质粒ｐＪＳＡ１１７５,置于痘苗病毒Ｐ７．５ｋ早／晚期启动子控制下。将此重组质粒用脂质体Ｌｉｐｏｆｅｃｔｉｎ方法转染已受野型ＴＫ￣＋痘苗病毒天坛株感染的ＴＫ￣－１４３细胞,通过同源重组机制和标志基因ＬａｃＺ产物的蓝斑显色作用,以及ＢｕｄＲ试剂对ＴＫ表型的选择压力,筛选出整合有ＨＳＶ－２ｇＤ基因的重组痘苗病毒。Ｓｏｕｔｈｅｍ杂交表明,ＨＳＶ－２ｇＤ基因已正确地插入痘苗病毒ＴＫ基因区内;间接免疫荧光检测显示,ＨＳＶ－２ｇＤ蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗ＨＳＶ－２ｇＤ中和抗体。用重组病毒免疫小鼠,３周后可使９４％（１７／１８）的小鼠对抗ＨＳＶ－２的致死量攻击,表明重组病毒具有明显的免疫保护作用。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.