人乳头瘤病毒11型L1主要衣壳蛋白的B细胞表位多肽作为疫苗的研究

分析了人乳头瘤病毒11型(HPV11)L1主要衣壳蛋白的B细胞优势表位,并以此为基础研制表位多肽疫苗。研究中采用Goldkey和．PC／Gene软件系统,分析HPV11的L1主要衣壳蛋白的二级结构、抗原性、B细胞表位,并引人氨基酸抗原性指数,综合评估其B细胞优势表位。Fmoc固相合成表位多肽,高效液相层析方法纯化,毛细管电泳分析其纯度。与0．2ml佐剂完全乳化后,按50μg／只的剂量免疫小鼠,进行动物水平的免疫效果评价。取免疫小鼠血清,与HPV11 DNA阳性的尖锐湿疣患者的疣体上清液结合,鉴定免疫后小鼠所产生抗体的特异性。发现HPV11的L1主要衣壳蛋白的第426～439位和第487～501位具有较高的免疫原性,可明显诱导小鼠血清抗体滴度升高,且该抗体与人尖锐湿疣的疣体组织上清液呈阳性反应。说明所选这两个肽段为HPV11的L1主要衣壳蛋白的B细胞优势表位,但是否具有功能特异性,尚需进一步研究。     

Biochemical and Molecular Characterization of Tetracycline-Resistant Aeromonas veronii Isolates from Catfish

Eighty-one tetracycline-resistant Aeromonas sp. strains were isolated from farm-raised catfish. Morphological and biochemical characteristics indicated that 23 of the 81 aeromonads were Aeromonas hydrophila, 7 isolates were Aeromonas trota, 6 isolates were Aeromonas caviae, 42 isolates were Aeromonas veronii, and 3 isolates were Aeromonas jandaei. However, the AluI and MboI restriction fragment length polymorphism (RFLP) patterns of the PCR-amplified 1.4-kb 16S rRNA gene from all 81 tetracycline-resistant aeromonads from catfish were identical to the RFLP banding patterns of A. veronii ATCC 35626, indicating that all 81 isolates were strains of A. veronii. A multiplex PCR assay successfully amplified the 5 tetracycline-resistant genes (tetA to E) from the genomic DNA of all 81 isolates. The assay determined that tetE was the dominant gene occurring in 73/81 (90.0%) of the aeromonads. Plasmids (2.0 to 20 kb) were isolated from 33 of the 81 isolates. Dendrogram analysis of the SpeI pulsed-field gel electrophoresis identified 15 distinct macrorestriction patterns among the isolates. Our results indicate the need for use of 16S rRNA in the identification of Aeromonas spp. and the prevalence of catfish as a reservoir of tet genes.     

雪莲PBP基因表达载体的构建

目的:利用新疆雪莲特殊功能基因磷脂酰乙醇胺结合蛋白基因(XLPBP)与基础质粒构建植物表达载体pXLPBP,为介导该基因在植物中表达,以期提高植物抗寒力的转基因研究打下基础。方法:利用设计好的两端加有EcoRⅠ酶切位点的引物,对XLPBP全长基因片段进行PCR扩增,获得700bp左右大小的片段,将其纯化回收并与同样经过EcoRⅠ酶切的质粒pCAMBIA3301连接;然后采用冻融法和电击法,将含XLPBP基因的载体pCAMBIA3301转入根癌农杆菌中。结果:通过一系列分子克隆方法获得含雪莲XLPBP基因的植物表达载体,并经PCR实验证实。结论:利用以自身携带的非编码区为调控序列的XLPBP全长基因和双向表达载体pCAMBIA3301为基础构建植物表达载体,可望提高外源基因表达量。     

贵州苏铁根的解剖学研究

对贵州苏铁（Cycas guizhouensis K.M.Lan et R.F.Zou）的根进行了解剖学研究,结果表明：（1）据外部形态和内部结构不同,贵州苏铁的根可分为正常根、珊瑚状根和肉质根三种类型。（2）正常根与肉质根的木栓形成层起源于表皮内方第1～2层皮层薄壁细胞,珊瑚状根的木栓形成层则起源于外方皮层的第2～3层细胞,而非中柱鞘。（3）首次发现肉质根一种新的增粗方式,是由周皮内方的皮层薄壁细胞恢复分裂能力,持续或周期性产生新的皮层薄壁细胞,引起根的增粗。     

Co-limitation of potato growth by Potato Cyst Nematode (Globodera rostochiensis) and Rhizoctonia solani

Globodera rostochiensis and Rhizoctonia solani are the most important growth limiting factors influencing potato production in Iran. The effects of inoculation with Potato Cyst Nematodes (PCN) (0, 50, 75 and 100 cysts/3.5?kg soil) and R. solani (with or without inoculation) on potato growth and development were investigated in cultivars Santé and Marfona. Inoculation with R. solani induced severe damage, especially when inoculation was accompanied with high density of PCN. The damage caused by R. solani tended to increase with an increase in PCN density, especially in Marfona. In Santé, number of stems or branches per plant significantly increased by inoculation with R. solani, while in Marfona it was significantly affected either by R. solani inoculation or PCN density. In Santé, number of stolons per plant was significantly increased by PCN, but not by R. solani. In Marfona, however, the number of stolons per plant was significantly affected either by R. solani inoculation or by presence of PCN, but not affected by PCN density. The general effect of R. solani or PCN inoculation treatments on shoot, below-ground and total dry weight of potato was significant, but strongly affected by cultivar. In general, our study supports the synergistic interaction between R. solani and PCN and its moderation by the use of a resistant cultivar such as Santé.     

Cytokine single nucleotide polymorphisms in patients’ with gallstone: dose TGF-β gene variants affect gallstone formation?

Gallstone is a common biliary disorder with several risk factors. Immune responses and inflammatory cytokines are important in this disease; as a result, some cytokines can be detected in bile fluid. In this research, cytokine gene polymorphisms were studied, and their effects on gallstone formation were evaluated. On 158 gallstone patients and 254 normal subjects, by PCR- RFLP method, IL-4-C590T polymorphism and by ARMS-PCR method, IFN-γ T+874A, TNF-α-A308G, IL-6 G-174C and TGF-β T+869C variants were studied. Pathologic evaluations were done on surgical specimens. There were no significant differences in distribution of evaluated polymorphisms between patient group and normal control group (P > 0.05), except TGF-β +869T allele (P = 0.04, OR = 1.23, 95 % CI = 1–1.79) which was higher in patients with gallstone. Although the pro-inflammatory cytokines such as TNF-α and IL-6 may promote gallstone formation, in this study no significant correlation between TNF-α and IL-6 polymorphisms and gallstone formation was seen. It is taught that TGF-β may affect gallbladder cells to promote gallstone formation and higher producer TGF-β +869T allele can be a risk factor of gallstone disease, so further studies would be more elucidative.     

Cytokine gene polymorphism and graft-versus-host disease: a survey in Iranian bone marrow transplanted patients

Graft versus host disease (GVHD) is a major complication of bone marrow transplantation (BMT). Numerous studies have shown the potential role of cytokine genotypes in the occurrence of GVHD. In this retrospective, case–control study we aimed to investigate the association between 13 cytokine genes and acute GVHD (aGVHD) after HLA-identical sibling BMT in 91 Iranian subjects. Negative association was found between aGVHD and donor IL-10/GCC haplotype or donor IL-4Ra-A allele in the population study. When compared within the leukemia subgroup, we observed positive association between recipient IL-1α ?889/C allele and aGVHD. Also there were negative association between recipient IL-10/CAA haplotype and donor IL-4Ra/A allele and development of aGVHD. Among the different genotypes only donor IL-4Ra and donor IL-12 showed significant association. We conclude that several cytokine polymorphisms are positively and negatively associated with aGVHD in Iranian HLA matched siblings, of which IL-4Ra and IL-12 may play important roles.     

Reaction of sunflower varieties to Sclerotinia sclerotiorum under various inoculation methods,time of field evaluation and phylogenic stages of host

Sunflower cultivation is affected seriously by Sclerotinia sclerotiorum (Lib.) de Bary in Iran, particularly north-western areas. Because of economic and environmental harms by chemical control, it is necessary to develop cultivars with adequate genetic resistance for reduction of yield losses. The purpose of this study was to find an effective method of inoculation with S. sclerotiorum under field evaluations. Three stem-inoculation techniques including: 1 – mycelium plug, 2 – oxalic acid solution (OAS) and 3 – infested wheat seeds with Sclerotinia mycelium were employed under field conditions. Four genotypes including Ghalami (local variety in market), Confeta, Allstar and Master were used in this study. The lesion length, lesion width and lesion as up and down leading on the stem from inoculation site were measured after 3, 7, 10 and 14 days of inoculation. The analysis of variance showed significant difference between all employed techniques and incubation days after inoculation. Mycelial plug (MP) inoculation technique produced significantly more developed lesions on the treated stems. In spite of this effect, Master variety demonstrated reasonable resistance reaction against the disease. The progress of disease in wounded treatments was also faster than the non-wounded ones. And, the shortest time to obtain significant differences between varieties was 10?days after inoculation. By comparison of results of lesion length at flowering and seed-filling stages, the more obvious effectiveness of the disease was observed at the second stage. Finally, there were negative correlations between mean temperature and mean lesion length in all three inoculation methods.     

Studies on the transmission of some Fusarium spp. to potato progeny tubers from mother plants and its biological control

Fusarium spp. attack potato roots causing root-rot, damping-off and wilt disease in Assuit Governorate. Forty-five Fusarium isolates were isolated from F. nygamai, F. acutatum, F. solani, F. proliferatum, F. subglutinans, and F. oxysporum. Isolates were tested for their pathogenic capability on Burn potato variety during growing season 2007/2008. Isolates infect potato plants causing either damping-off or wilt symptoms. Isolates varied in their virulence. Role of potato tuber seed in the transmission of the causal pathogen to daughter using Electrophoresis. Protein profiles of the tested isolates divided into four sub-clusters at similarity levels 93.79, 91.55 and 92.62% while isolate of Fusarium profile No. 11 formed separate sub-clusters at similarity level 69.79%. F. nygamai and F. solani were notable exception because profile No. 4 of F. nygamai from roots and profile No. 4 from sprouts were almost identical (similarity level 96.81%); similarity level between profile No. 8 from roots and profile no/8 from sprouts was 95.44%. Results prove that F. nygamai and F. solani are potato tuber seed-borne fungus. T. harzianum, T. viride, T. longibrachiatum, G. virens and E. nigrum or its filtrate inhibited the growth of F. nygamai, F. acutatum, F. solani, F. proliferatum, F. subglutinans and F. oxysporum. The formulation of T. harzianum, T. longibrachiatum and G. virens against tested pathogenic fungi reduce disease incidence under greenhouse conditions.     

Proteome Variations in Pancreatic Stellate Cells upon Stimulation with Proinflammatory Factors

Pancreatic stellate cells are key mediators in chronic pancreatitis and play a central role in the development of pancreatic fibrosis, stromal formation, and progression of pancreatic cancer. This study was aimed at investigating molecular changes at the level of the proteome that are associated with the activation of pancreatic stellate cells by proinflammatory factors, namely TNF-α, FGF2, IL6, and chemokine (C-C motif) ligand 4 (CCL4). They were added individually to cells growing in serum-free medium next to controls in medium supplemented with serum, thus containing a mixture of them all, or in serum-free medium alone. Variations were detected by means of a microarray of 810 antibodies targeting relevant proteins. All tested factors triggered increased proliferation and migration. Further analysis showed that TNF-α is the prime factor responsible for the activation of pancreatic stellate cells. CCL4 is associated with cellular neovascularization, whereas FGF2 and IL6 induction led to better cellular survival and decreased apoptotic activity of the stellate cells. The identified direct effects of individual cytokines on human pancreatic stellate cells provide new insights about their contribution to pancreatic cancer promotion.