Highly Efficient and Economical Baculovirus Expression System for Preparing Human Papillomavirus Type16 Virus-like Particle

Human papillomavirus (HPV) is the most importantagent of human primary cervical cancer, and the list ofmalignancies caused by or associated with HPV is stillgrowing. HPV has been detected in carcinomas of larynx,esophagus, oral cavity, nasal sinus, skin, …     

Analysis of part of the Trichophyton rubrum ESTs

Superficial fungi are the etiologic pathogens of various dermatophytoses, such as tinea capitis, tinea corporis, tinea inguinalis, tinea manus, tinea unguium and tinea pedis. These widespread infections affect up to 25% of the worlds population, a percentage that has continued to increase in recent years. T. rubrum is the most common superficial fungus, accounting for at least 60% of all infections of this type. T. rubrum can induce dermatophytoses in different parts of the hu-man skin, and ca…     

Analysis of ESTs and gene expression patterns of the poste-rior silkgland in the fifth instar larvae of silkworm, Bom-byx mori L.

The fibroin gene expression pattern and regulation of the posterior silkgland were studied by means of expressed sequence tags (ESTs) using the first and fifth day larvae of the fifth instar of silkworm, Bombyx mori L (strain: C 108). The results showed that there were 911 repetitive ESTs and 1950 single sequences (Singlets) among total 2861 consentient sequences, which were spliced. 1335 sequences were identified and the other 1526 were unknown. 5560 sequences (55.89%) in the posterior silkgland cell of the silkworm were new ESTs without ho-mology with EST data published by Mita et al. The number of repetitive ESTs and single sequences from the first day larvae of the fifth instar was double more than that of the fifth day of the same instar in the silkworms. The unigenes which were more than 50 in repetitive EST size (contig size) came to only about 0.5% in total consentient sequences. There were significant differences between gene expression frequencies, and expressed genes were related to fibroin synthesis and its secretion and fibroin composition. Comparing the fifth day with the first day of the fifth instar, the genes-expressed quantity of fibroin heavy-chain gene was 18 fold higher, fibroin light-chain gene 9 fold and fibroin P52 gene 8 fold. 508 genes functioned for cellular component and 315 for enzyme after function tracing. These results implied that the gene expression of the first day was mainly for preparation for fibroin synthesis except for the growth of silkgland cells, and the gene expression of the fifth day of the fifth instar was mainly for synthesizing and excreting fibroin. Because the ratio of heavy chain, light chain and p25 of fibroin was not 6:6:1 as theoretically expected, or its special H-chain structure, the H-chain gene was not easy to detect through EST technique. Most of genes among total 2861 consentient sequences functioned for fibroin synthesis and secretion. This suggested the fibroin synthesis and secretion procedure of the posterior silkgland was more complex than the knowledge we have.     

贵州苗族新生儿HbF中Gγ/Aγ比值测定与AγT基因频率分析

用含Triton X-100的酸性尿素聚丙烯酰胺凝胶电泳(PAGE)法测定了贵州省108例苗族新生儿脐带血胎儿血红蛋白(HbF)中Gγ/Aγ值.显示107例(占99.07%)新生儿的Gγ值在55-78%之间,均值与标准差为70.42±3.45;高Gγ值者1例(占0.925%);低Gγ值者未发现。用Large-pore Vydac C4柱反相高效液相层析法对108例样本进行了AγT基因频率（fAγT）分析,fAγT为0.0648。 The ratio of Gγ to Aγ of fetal hemoglobin inn 108 cord blood samples of Miao nationality newborns in Guizhou province was determined by the polyacrylamide gel electrophoresis(PAGE).The result showed that one case had high Gγ value(0.925%) and 107 babies the normal Gγ values of 55 to 78%(70.42+3.45)。The AγT gene frequency(fAγT) analysis in 108 samples was done by high pressure liquid chromatography using a large pore Vydac C4 column,the fAγT was 0.0648.     

The diversity effect of inductive reasoning under segment manipulation of complex cognition

Forming a general reasonable conclusion from a limited number of examples is one of the most mar-velous psychological activities of human beings. This kind of activities is inductive reasoning which is a key issue rising from the cross fields of psychology, phi-losophy, cognitive science, and so on. It is a thinking process of generalizing from concrete cases. The di-versity effect of inductive reasoning refers to the phe-nomenon that the inductive argument of more diverse premise can obtain s…     

The Comparative and Functional Study between Two Construction Methods of shRNA Expression Vector Targeted LMP1 Gene Encoded by EBV

To look for a more stable and convenient way of constructing short hairpin RNA expression vectors targeting the latent membrane protein-1(LMP-1)encoded by Epstein-Barr virus(pshLMP1),and to study the inhibition function of pshLMP1 expression vectors in HNE1 cells,we designed the pshLMP1 expression cassette and pshLMP1 expression vectors by both the annealing method and PCR method and then co-transfected with pEGFP-N1-1158 into HNE1 cells to observe the mRNA and protein levels of LMP-1 genes by green fluorescence analysis,RT-PCR and western blot.pshLMP1 expression vectors were successfully obtained by both methods but better cloning efficiency was achieved and fewer deletions and mutations of nucleotides were achieved with the PCR method.Furthermore,the mRNA and protein levels of LMP-1 genes were down-regulated by pshLMP1 expression vectors.According to our research,we found that the PCR method provides a more efficient way to construct pshLMP1 expression vectors which have the ability to inhibit the function of LMP-1 genes expressed in HNE1 cells,and also provides a novel application of RNA interference technology against-EBV.     

肉桂酸衍生物对铜绿假单胞菌PAO1 III型分泌系统的影响

【目的】铜绿假单胞菌是引起医院获得性感染最常见的条件致病菌,而III型分泌系统(Type III secretion system,TTSS)是其致病的主要因子之一。本文从合成的21个肉桂酸衍生物中筛选影响TTSS效应子(Effector)产生的化合物,并初步研究其作用机制。【方法】将TTSS效应子合成基因exoS的转录报告质粒pAT-exoS转入菌株PAO1中,获得PAO1(pAT-exoS)。待筛选的化合物与PAO1(pAT-exoS)菌株共培养6 h后,检测exoS基因的表达,从中筛选影响exoS基因表达的化合物。【结果】筛选结果表明：21个化合物中,3个化合物抑制exoS基因表达,2个化合物则促进exoS基因表达。此外,化合物TS128、TS143和TS160对菌株生长有明显的抑制作用。Western blot实验进一步证实筛选得到的化合物TS108、TS128和TS165可抑制ExoS的产生;化合物TS139和TS143则促进ExoS的产生。为进一步研究抑制剂的作用机理,过量表达TTSS主要的调控因子exsA基因可部分消除抑制剂TS108和TS165的抑制效果;而rsmZ rsmY双基因突变体PAO6421中添加抑制剂TS108和TS165并不能显著抑制exoS基因的表达,同样,抑制剂TS108和TS165也不影响受Gac/Rsm信号传导系统调控的群体感应信号分子的产生。【结论】抑制剂TS108和TS165的作用机制可能主要是影响esxA基因,从而影响exoS基因表达及蛋白产量。