Allele imputation for the killer cell immunoglobulin-like receptor KIR3DL1/S1

Highly polymorphic interaction of KIR3DL1 and KIR3DS1 with HLA class I ligands modulates the effector functions of natural killer (NK) cells and some T cells. This genetically determined diversity affects severity of infections, immune-mediated diseases, and some cancers, and impacts the course of immunotherapies, including transplantation. KIR3DL1 is an inhibitory receptor, and KIR3DS1 is an activating receptor encoded by the KIR3DL1/S1 gene that has more than 200 diverse and divergent alleles. Determination of KIR3DL1/S1 genotypes for medical application is hampered by complex sequence and structural variation, requiring targeted approaches to generate and analyze high-resolution allele data. To overcome these obstacles, we developed and optimized a model for imputing KIR3DL1/S1 alleles at high-resolution from whole-genome SNP data. We designed the model to represent a substantial component of human genetic diversity. Our Global imputation model is effective at genotyping KIR3DL1/S1 alleles with an accuracy ranging from 88% in Africans to 97% in East Asians, with mean specificity of 99% and sensitivity of 95% for alleles >1% frequency. We used the established algorithm of the HIBAG program, in a modification named Pulling Out Natural killer cell Genomics (PONG). Because HIBAG was designed to impute HLA alleles also from whole-genome SNP data, PONG allows combinatorial diversity of KIR3DL1/S1 with HLA-A and -B to be analyzed using complementary techniques on a single data source. The use of PONG thus negates the need for targeted sequencing data in very large-scale association studies where such methods might not be tractable.     

三峡库区橙汁加工品种的筛选与配套研究

为满足三峡库区橙汁加工对甜橙品种熟期配套和品质调控的需要,通过对引进的特早熟品种早金(Early Gold)、早熟夏橙品种蜜奈(Midknight)和晚熟夏橙品种奥林达(Olinda),以及从本地主栽甜橙品种锦橙中选出的中晚熟品种晚熟锦橙的品质和熟期分析,初步证明这几个品种与本地主栽品种锦橙配套,可以实现从11月至次年6月的8个月成熟期配套,基本满足橙汁加工厂周年生产的需要。筛选出的长叶橙、宜圆3号等国内品种和从国外引进的埃及糖橙、早金等品种,其果汁具有较高的含糖量和固酸比,可用于调配降低三峡库区橙汁的酸度,改善果汁风味。     

白囊耙齿菌子实体提取物对小鼠慢性肾小球肾炎的预防与治疗

本文对白囊耙齿菌子实体不同提取物防治慢性肾小球肾炎进行研究,采用腺嘌呤灌胃法建立小鼠慢性肾小球肾炎模型,探究白囊耙齿菌子实体水提取物和醇提取物的高、中、低剂量组对慢性肾小球肾炎的预防作用及不同提取物的高剂量组对慢性肾小球肾炎的治疗作用,并对小鼠体重进行测量,检测尿蛋白(UP)、尿素氮(BUN)、肌酐(Cr)、总胆固醇(TC)、超氧歧化酶(SOD)、一氧化氮(NO)、丙二醛(MDA)、白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)指标,分析肾、肝、脾、胸腺的病理变化。结果表明,水提取物中剂量(240mg/kg)能显著降低小鼠尿蛋白含量和肾组织中炎症因子含量,使血清生化指标维持正常,并对免疫器官有保护作用;醇提取物高剂量(200mg/kg)对慢性肾小球肾炎有治疗作用,但对免疫器官保护作用较弱。研究表明,白囊耙齿菌子实体水提取物中剂量具有预防慢性肾小球肾炎的作用且效果优于阳性药肾炎康胶囊,具有开发利用潜力,机制有待于进一步研究。     

大肠杆菌发酵生产重组人源抗-HBs Fab

在KLF2000发酵罐中利用补料分批培养技术培养表达含重组质粒pBAD/HBs Fab的TOP10大肠杆菌,生产人源抗-HBs Fab,为批量生产作准备,在发酵过程中,控制溶氧30%以上,温度37℃,在基础培养基内生长4h后,补加以甘油为碳源的补料,继续生长到9h,加入阿拉伯糖,至终浓度为0.02%,30℃诱导表达5h,收集菌体,纯化制备目的蛋白。利用Western blot方法检测Fab抗原性,Dot blot方法检测生物学活性。14h发酵结束后,菌体密度最终达96g/L,纯化所得蛋白大约占菌体总蛋白的6%,含量为80mg/L,以重组质粒pBAD/HBs Fab,大肠杆菌TOP10表达表达比率与摇瓶相比没有降低,表达量达80mg/L左右,为大批量生产作了准备。     

Long‐term effects of vegetational restoration on soil microbial communities on the Loess Plateau of China

Soil microbial communities are important indicators for evaluating the effectiveness of vegetational restoration. The ability of soil microbial communities to recover under six types of restoration was examined using Biolog, phospholipid fatty acid (PLFA), and denaturing gradient gel electrophoresis (DGGE). Soil samples were collected from eroded loessial soils that had been restored for 30 years on the Loess Plateau with natural grass, black locust, korshinsk peashrub, Chinese pine, and mixed forest of Chinese pine‐indigo and black locust‐indigo. Part of sloped farmland that had not been restored represented pre‐restoration conditions. An 80‐year‐old forest of Chinese arborvitae represented the end point of restoration. Soils that were originally farmed but have been under restoration treatments for the past 30 years had significantly higher average well‐color development, total PLFAs, bacterial PLFAs, fungal PLFAs, and Shannon indices of catabolic diversity, structural diversity, and bacterial phylogenetic diversity. These indicators, though, were lower than those for the native forest. Principal component analysis significantly separated the sloped farmland, Chinese arborvitae, and the six types of restoration. Redundancy analysis indicated that pH, total organic carbon, total nitrogen, available nitrogen, and available phosphorus were the most important environment factors in affecting soil functional diversity, and C/N ratio and available nutrients were the main properties in determining microbial community structure. Vegetational restoration substantially increased microbial biomass and microbial physiological activity, and shifted microbial community structure, but the community structures and compositions did not recover to the status of the natural forest during the early stage (<30 years) of vegetational restoration.     

不同供水下白羊草(Bothriochloa ischaemum)离体根呼吸特征——基于稳定碳同位素示踪技术

植物根呼吸是土壤呼吸的主要组成,研究根呼吸对生态系统碳收支及碳平衡有重要意义。采用~(13)C脉冲标记技术,在3种供水条件下,对比不同根离体时间(标记后0,6,24,48,216,360 h)的白羊草离体根呼吸速率和根呼吸释放的δ~(13)C同位素比值变化,分析根参数与离体根呼吸相关性。结果表明:1)不同离体时间的离体根呼吸速率变化趋势一致,3种供水条件下无显著差异,均在0—20 min急剧下降,下降范围为32%—39%。2)测定离体根呼吸释放的δ~(13)C在不同离体时间的变化,为实时监测转移到白羊草根系的~(13)CO_2在根部释放的过程提供了新思路;不同离体时间,3种供水条件下根呼吸释放的δ~(13)C在2 h内均值大小呈:供水充分轻度胁迫重度胁迫。随离体时间(0—360 h)推移根呼吸释放的δ~(13)C均值先增大后减小,在216 h达到峰值31.46‰;3)离体根呼吸速率和根呼吸释放的δ~(13)C受根系根面积、比根面积、N含量、C/N及根组织δ~(13)C的影响显著。4)轻度水分胁迫可促使根系生长(C固定)和根呼吸(C代谢)同时增加。     

放牧后羊草和芦苇叶组织转化的比较

运用组织转化理论与分析方法,对羊草与芦苇在放牧后的叶组织转化进行了比较研究。结果表明,整个生长季内,除5月份外,芦苇的绿叶数大于羊草,羊草的绿叶长大于芦苇;2种禾草的叶片发生率和伸长率均在生长初期最大,以后逐渐降低,萎蔫率正好相反;芦苇叶片寿命短,叶组织转化快、叶组织物质积累呈抛物线型增长,如果不及时利用,会造成资源的浪费,羊草叶片寿命较长,叶组织转化慢。可以通过不同的利用方式和时间来改善羊草草地的质量。     

冠状病毒HcoV-229E S1蛋白的原核表达及鉴定

目的克隆表达冠状病毒HcoV-229E S1基因片段,表达S1蛋白。方法合成冠状病毒HcoV-229ES1蛋白特异性基因片段并克隆入pET21a原核表达载体,转化BL21（DE3）菌,经IPTG高效诱导表达得到重组蛋白,用金属螯合亲和层析纯化,并通过Western blot对表达的重组蛋白进行鉴定。结果获得了主要以包涵体形式存在的目的蛋白,Western blot鉴定其为S1基因片段蛋白。结论成功构建了HcoV-229E S1蛋白的表达载体,并在BL21（DE3）中得到了高效表达,为下一步表达蛋白免疫原性及疫苗抗病毒保护性测定打下了基础。     

SGA微囊制备及其与大鼠胰岛生物相容性的体外观测

目的:制备硫酸酯化葡甘聚糖-海藻酸钡(SGA)微囊,包裹大鼠胰岛,对比观测SGA微囊的体外生物相容性.方法:溶液共混法制备硫酸酯化葡甘聚糖-海藻酸盐混合凝胶液,改良Omer法制作SGA微囊.X射线衍射分析SGA二元共混膜结构;SGA微囊化SD大鼠胰岛,体外培养14d后,胰岛素刺激释放实验对比SGA微囊和APA、ABa(海藻酸钡)微囊对大鼠胰岛的影响.结果:衍射图谱显示硫酸酯化葡甘聚糖-海藻酸钡共混膜在原属于海藻酸钡的2θ 11°、20.3°结晶峰变小,在20 29.8.出现新结晶峰,说明SGA二元共混膜相互作用强烈,融合较好;胰岛素刺激释放实验显示,体外培养后SGA微囊对大鼠胰岛的影响与APA、ABa微囊无差别(P＞0.05).结论:SGA微囊膜结构稳定,融合度较好,对大鼠胰岛体外生物相容性较高,可以为下一步体内移植所应用.