华南主要树木丛枝菌根真菌物种多样性调查研究

为调查华南地区丛枝菌根（arbuscular mycorrhiza,AM）真菌的物种多样性与资源分布状况,本研究对该区域红花羊蹄甲（Bauhinia blakeana）、黄梁木（Neolamarckia cadamba）、构树（Broussonetia papyrifera）、尾叶桉（Eucalyptus urophylla）和杧果（Mangifera indica）5种主要树木AM真菌侵染状况进行研究,通过形态学特征及核糖体18S rRNA基因序列分析鉴定AM真菌种类。结果表明:1）5种树木均能形成丛枝菌根,红花羊蹄甲和尾叶桉为疆南星型（Arum-type）,杧果、构树和黄梁木为重楼型（Paris-type）。红花羊蹄甲和杧果的菌根侵染率高、孢子密度大,构树、黄梁木和尾叶桉的AM真菌侵染率和孢子密度相对较低。2）鉴定出AM真菌5属8种,分别为无梗囊霉属（Acaulospora）的浅窝无梗囊霉（Acaulospora lacunosa）和刺无梗囊霉（Acaulospora spinosa）,斗管囊霉属（Funneliformis）的摩西斗管囊霉（Funneliformis mosseae）,根孢囊霉属（Rhizophagus）的根内根孢囊霉（Rhizophagus intraradices）,以及球囊霉属（Glomus）的3种Glomus spp.和硬囊霉属（Sclerocystis）1种Sclerocystis sp.。3）球囊霉属的AM真菌广泛分布在红花羊蹄甲、杧果和构树根际,刺无梗囊霉分布在杧果和尾叶桉根际,摩西斗管囊霉为优势种,分布在构树和黄梁木根际;而浅窝无梗囊霉和硬囊霉只分布在1种树根际,表明其宿主专一性相对较强。结果显示华南树木根际土壤中AM真菌物种多样性较高,同时本研究为深入研究该地区林木根际AM真菌功能多样性提供了初步科学依据。     

Differential Expression Analysis of MUC4 and MUC13 Genes Between Resistant and Sensitive Weaned Meishan Piglets to ETEC F18

MUC4 and MUC13 genes as important candidate genes for enterotoxigenic Escherichia coil (ETEC) F4 resistance,may play an important role in the process of against ETEC F18 infection in weaned piglets. In this study,ETEC F18-resistant and -sensitive weaned Meishan piglets were used,and the expression levels of MUC4 and MUC13 genes in 11 tissues (heart,liver,spleen,lung,kidney,stomach,muscle,thymus,lymph nodes,duodenum and jejunum) were determined by quantitative Real-time PCR. The results showed that MUC4 and MUC13 genes were broadly expressed with different expression levels in all the 11 tissues. In the thymus and lymph tissues,the expression of MUC4 gene in resistant piglets was significantly higher than that in sensitive piglets (P<0.05);In the lung tissue,theMUC13 gene expression level in resistant individuals was significantly higher than that in sensitive individuals (P<0.05),and in the intestinal tissues of duodenum and jejunum, the expression level of MUC13 gene was relatively higher in resistant individuals. Thus we speculated that the high expression of MUC4 gene in immune tissues and MUC13 gene in intestinal tissues might improve the immune ability of piglets,protect and lubricate the intestinal tract, and resist ETEC F18 infection.     

Prevalence of Cryptosporidium species isolated from HIV/AIDS patients in southwest of Iran

This study aimed to determine the prevalence and species of Cryptosporidium among HIV/AIDS patients in southwest of Iran. Two hundred fifty faecal samples from HIV patients were examined for the presence of Cryptosporidium oocysts using a conventional coproscopic approach. Such oocysts were detected in 18 (7.2%) out of 250 faecal samples. Genomic DNAs from 250 samples were then subjected to a nested-PCR-RFLP technique targeting different loci of 18S rRNA gene for species identification. Out of 250 samples, 27 (10.8%) were positive for different Cryptosporidium spp; Restriction patterns resulting from the digestion of the nested amplicon with restriction endonucleases VspI and SspI showed that C. parvum (70.38%) was the most prevalent species, followed by C. hominis (25.92%) and C. meleagridis (3.7%), respectively. The mean CD4+ T-cell count was 215 cells/μL. There was a strong association between cryptosporidiosis and CD4+ T-cell count (P = 0.000) with the highest prevalence recorded among patients with CD4+ T-cell count < 200 cells/μL. This confirms that there is a low opportunity for this parasite to get established as the patients CD4+ T-cell count increases. Also HIV infection increased the risk of having Cryptosporidium. Our epidemiological findings are useful for any preventive intervention to control disease diffusion.     

Characteristics of 18F-FDG and 18F-FDOPA PET in an 8-year-old neutered male Yorkshire Terrier dog with glioma: long-term chemotherapy using hydroxyurea plus imatinib with prednisolone and immunoreactivity for PDGFR-β and LAT1

An 8-year-old neutered male Yorkshire Terrier dog presented with head pressing, vestibular ataxia, neck tenderness, and no oculocephalic reflex. A demarcated lesion in the pons was identified on MRI. The patient was tentatively diagnosed with a glioma and was treated with hydroxyurea plus imatinib and prednisolone. After 30 days of therapeutic treatment, the patient showed a clear improvement in neurological signs, which lasted for 1117 days. On day 569 after the initiation of treatment, ¹⁸F-fluorodeoxyglucose (FDG)-positron emission tomography (PET) was performed with no significant findings on visual analysis. The average and maximal standardized uptake values (SUVs) were 1.92 and 2.29, respectively. The tumor-to-normal-tissue (T/N) ratio was 0.97. The first evidence of clinical deterioration was noticed on day 1147. On day 1155, 3,4-dihydroxy-6-[¹⁸F]-fluoro-l-phenylalanine (¹⁸F-FDOPA)-PET was performed. High uptake of ¹⁸F-FDOPA was observed in the intracranial lesion. The mean and maximal SUVs of the tumor were 1.59 and 2.29, respectively. The T/N ratio was 2.22. The patient was euthanized on day 1155 and histopathologic evaluations confirmed glioma (astrocytoma). This case shows that chemotherapy with hydroxyurea plus imatinib may be considered in the treatment of canine glioma. Furthermore, this is the first case describing the application of ¹⁸F-FDG and ¹⁸F-FDOPA in a dog with glioma.     

山羊子宫内膜基质细胞对上皮细胞IL-18分泌活性的调节作用

为了探究性腺激素作用下山羊子宫内膜上皮细胞（EEC）中白细胞介素-18（IL-18）的分泌活性,以及子宫内膜基质细胞（ESC）对其分泌活性的调节作用。基于套皿共培养永生化山羊EEC和ESC为体外研究模型,通过ELISA和Western blot方法检测E2和/或P4对EEC中IL-18分泌水平的作用以及ESC对该作用的影响。ELISA结果显示,E2较无激素对照组可显著增强单独培养EEC中IL-18的分泌水平（P〈0.05）,P4单独或与E2联合作用则进一步增强其分泌活性（P〈0.01）;EEC与ESC共培养模式下,ESC细胞对性激素作用下EEC中IL-18分泌活性具有显著的抑制作用。Western blot结果显示,在单独培养或与ESC共培养模式下,EEC培养上清液中IL-18均以相对分子质量大小18 300的具有生物活性的蛋白形式存在。本研究结果表明,山羊ESC对性腺激素作用下EEC中IL-18的分泌活性具有重要调节作用。     

The influence of soil properties on the structure of bacterial and fungal communities across land-use types

Land-use change can have significant impacts on soil conditions and microbial communities are likely to respond to these changes. However, such responses are poorly characterized as few studies have examined how specific changes in edaphic characteristics do, or do not, influence the composition of soil bacterial and fungal communities across land-use types. Soil samples were collected from four replicated (n = 3) land-use types (hardwood and pine forests, cultivated and livestock pasture lands) in the southeastern US to assess the effects of land-use change on microbial community structure and distribution. We used quantitative PCR to estimate bacterial–fungal ratios and clone libraries targeting small-subunit rRNA genes to independently characterize the bacterial and fungal communities. Although some soil properties (soil texture and nutrient status) did significantly differ across land-use types, other edaphic factors (e.g., pH) did not vary consistently with land-use. Bacterial–fungal ratios were not significantly different across the land-uses and distinct land-use types did not necessarily harbor distinct soil fungal or bacterial communities. Rather, the composition of bacterial and fungal communities was most strongly correlated with specific soil properties. Soil pH was the best predictor of bacterial community composition across this landscape while fungal community composition was most closely associated with changes in soil nutrient status. Together these results suggest that specific changes in edaphic properties, not necessarily land-use type itself, may best predict shifts in microbial community composition across a given landscape. In addition, our results demonstrate the utility of using sequence-based approaches to concurrently analyze bacterial and fungal communities as such analyses provide detailed phylogenetic information on individual communities and permit the robust assessment of the biogeographical patterns exhibited by soil microbial communities.     

NDV F与ChIL-18融合基因"自杀性"质粒的构建

根据GeneBank公布的鸡新城疫病毒(NDV)Lasota株F基因和鸡源性白细胞介素18(ChIL-18)基因序列分别设计特异性引物,且在鸡NDV F基因引物上游引入BamH Ⅰ位点、下游引入Sal Ⅰ位点,在Ch IL-18基因引物上游引入Sal Ⅰ位点、下游引入Hind Ⅲ、BamH Ⅰ位点.以鸡新城疫病毒Lasota株RNA为模板RT-PCR扩增其F基因,将F基因插入pMD18-T的BamH Ⅰ与Sal Ⅰ位点中.同时扩增上游带有Sal Ⅰ位点、下游带有Hind Ⅲ、BamH Ⅰ位点的ChIL-18基因,并将其插入到含有F基因的pMD18-T质粒的Sal Ⅰ、Hind Ⅲ位点之间,从而使NDV F基因与ChIL-18基因形成具有一个完整阅读框的融合基因,然后将融合基因片段定向克隆到pSFV单一启动子(CMV)下游,构建成F-ChIL-18融合基因重组"自杀性"质粒pSFV-F-ChIL-18.     

水稻新品种"吉农大18号"选育及栽培技术

水稻新品种“吉农大18号”是以组培7为母本，以93-14为父本进行有性杂交，经系谱法选育而成。3年吉林省区域试验产量结果比对照(通35)增产5．23％，1年生产试验产量比对照增产6．0％。“吉农大18号”具有高产、优质、抗性强、适应性广等特点，于2003年通过吉林省农作物品种审定委员会审定。     

猪带绦虫不同阶段45W-4BX和18kD基因联合表达及保护性分析

【目的】获得不同发育阶段基因联合表达的具有良好免疫保护性的重组抗原，为研制高效的猪囊虫基因工程疫苗奠定基础。【方法】PCR扩增截去45W-4B基因的信号肽和C端17个疏水氨基酸序列，经BamH I和EcoR I酶切后与表达载体pGEX－4T-1连接转化BL21感受态细胞，酶切及PCR扩增鉴定阳性克隆。测序正确的质粒经EcoR I和Not I酶切处理后与截去信号肽的18 kD基因连接，构建双基因融合表达载体pGEX-4BX/18。用IPTG诱导的表达产物，进行SDS－PAGE电泳、Western blot分析活性。分别用300μg重组GST-4BX、GST-4BX/18蛋白免疫猪，间接ELISA测定抗体水平。感染后90 d剖检计算各组的减虫率，比较评价重组抗原的免疫保护性。【结果】4BX/18 kD在大肠杆菌中获得高效表达，表达产物为50 kD的融合蛋白，并能被人囊虫和感染初期的猪囊虫阳性血清所识别。重组抗原免疫猪后45 d抗体达到峰值，联合表达重组抗原的减虫率为97%，GST-4BX免疫组的减虫率为95%。【结论】重组抗原4BX 和4BX/18kD均具有较好的免疫保护效果，有望利用它们研制出抗猪囊尾蚴病的高效疫苗。     

多头带绦虫Tm16与Tm18抗原基因的克隆及原核表达

为原核表达Tml6和Tm18重组蛋白,本研究以自然感染羊源脑多头坳原头节基因组DNA为模板分别扩增Tml6和Tm 18杭原基因全基因片段,测序鉴定后合成其开放阅读框架DNA片段,将基因组序列中的内含子去除并对其稀有密码子进行改造优化,构建重组表达质粒pGEX-Tm 16和pGEX-Tml8.转化大肠杆菌BL21后以IPTG诱导表达,SDS-PAGE分析表达产物并进行纯化.结果显示:在大肠杆菌中表达出带有GST标签的大小约为39.6 ku和39.4 ku的重组蛋白,经谷胱甘肤琼脂糖树脂纯化得到高纯度的可溶性的GST-Tm 16及GST-Tml8重组蛋白,western blot分析表明重组蛋白均能够被兔抗GST单克隆杭体特异性识别.