铃兰的胚胎学研究

铃兰（Ｃｏｎｖａｌｌａｒｉａ ｍａｊａｌｉｓＬ．）的花药四室,药壁发育为单子叶型。腺质绒毡层发育后期出现双核至多核。小孢子四分体主要呈左右对称型,偶见四面体型。胞质分裂为连续型。二细胞型花粉。子房三室,中轴胎座。胚珠倒生,双珠被,厚珠心。珠孔由内珠被形成。胚囊发育为葱型和英地百合型。受精以后,在子房壁、珠柄和外珠被细胞中含有草酸钙针晶。胚发育类似于石竹型,但基细胞发生纵裂形成两个子细胞。核型胚乳;合点端具巨形胚乳核     

人RHD基因的克隆和鉴定

目的克隆人RHD基因,并对其进行鉴定。方法以RhD阳性志愿者骨髓为材料,用TRIzol试剂提取总RNA;设计、合成人RHD基因扩增引物,RT-PCR方法扩增RHD基因片段;T／A克隆后将其亚克隆人pET28a（＋）载体中,经酶切、PCR和测序对重组质粒进行鉴定。结果骨髓总RNA被成功提取;RT-PCR成功扩增出RHD基因片段,其大小与预期约509bp基本一致;T／A克隆后再将其亚克隆,通过酶切和PCR证明RHD基因成功亚克隆入pET28a（＋）载体中;基因测序结果比对显示,与已公布的RHD基因（GenBank登录号为NM016124）序列基本一致,同源性为98％。结论成功克隆了RHD基因,这将为进一步研究奠定基础。     

逆相蒸发法制备茶多酚脂质体及质量评价

采用逆相蒸发法制备茶多酚脂质体并进行质量评价。通过二次回归旋转组合设计优化茶多酚脂质体制备工艺及配方,对其形态、结构、粒径分布等性质进行考察。研究结果表明,最佳配方为m（大豆卵磷脂）：m（胆固醇）=3：1、茶多酚质量浓度为7mg／mL、V（有机相）：V（水相）=4：1、磷酸盐缓冲液浓度15mmoL／L,此条件下包封率为50．37％;所制备的茶多酚脂质体形态呈圆球形或椭球形,为大单室脂质体,有效粒径为165．3nm,Zeta电位为-69．3mV。逆相蒸发法制备茶多酚脂质体方法简单可行,所制备的脂质体具有一定缓释性。     

Cadaveric study of the arterial anatomy of the upper lip

Arterial distribution of the upper lip was investigated in this study. The location, course, length, and diameter of the superior labial artery and its alar and septal branches were determined on 14 preserved cadaver heads. Another cadaver head was used to show the arterial tree by the colored silicone injection technique. The superior labial artery was the main artery of the upper lip and always originated from the facial artery. The superior labial artery was 45.4 mm in length, with a range from 29 to 85 mm. The mean distance of the origin of the superior labial artery from the labial commissura was 12.1 mm. The superior labial artery was 1.3 mm in external diameter at its origin. The mean distance of origin of the superior labial artery from the lower border of the mandible was 46.4 mm. The alar division of the superior labial artery was mostly found as a single branch (82 percent). Its mean length was 14.8 mm and the mean diameter at the origin was 0.5 mm. The distance between the origins of the superior labial artery and the septal branch was 33.3 mm. The septal branch was single in most of the cases (90 percent). The mean length of the septal branch was 18.0 mm and the diameter at its origin was 0.9 mm. After all dissections, it was concluded that the arterial distribution of the upper lip was not constant. The superior labial artery can occur in different locations unilaterally and bilaterally, with the branches showing variability.     

内蒙古地区刺叶柄棘豆(Oxytropis aciphylla Ledeb.)种群遗传多样性分析

用ISSR分子标记对内蒙古地区刺叶柄棘豆(Oxytorpis aciphylla Ledeb.) 5个地理种群进行了种群遗传多样性分析。结果表明：刺叶柄棘豆种群具有较高的遗传多样性,11个ISSR引物扩增出215条带,总的多态位点百分率为98.14%,Shannon多样性指数I=0.2108,Nei基因多样性指数 H=0.341 6,种内总基因多样性(H_t) 为0.2108,种群内基因多样性(H_s)为0.160 4, 大部分遗传变异(76.11%)的遗传变异存在于种群内, 23.89%的遗传变异存在于种群间。遗传分化系数(Gst)为0.238 9,基因流(Nm)为1.592 9。5个居群间已有遗传分化趋势,遗传漂变不会引起遗传分化。UPGMA遗传距离聚类结果表明, 5个地理种群中,植被类型为荒漠草原的4个种群之间遗传距离较近,与1个荒漠种群距离较远。     

穴居狼蛛毒中一个抗菌活性多肽的鉴定和纯化

从我国新疆地区产穴居狼蛛的毒液中分离纯化了一种多肽——狼蛛抗菌肽(Lycosin)。穴居狼蛛的粗毒在酸性聚丙烯酰胺凝胶电泳中可分出11条蛋白带,用0.6％的大肠杆菌琼脂胶覆盖在凝胶上进行鉴定表明,电泳迁移率最快的区带有抗菌活性。经鉴定该多肽分子系由43至45个氨基酸残基组成,N-末端为丙氨酸,分子中的碱性和疏水性氨基酸分别占总氨基酸残基数的1/4和1/3。     

Flower-inducing and -inhibiting activities of Phloem Exudate from Cotyledons of Pharbitis Seedlings

The flower-inducing and -inhibiting activities of phloem exudate (PE) prepared from cotyledons of Pharbitis seedlings were examined, using apex cultures in vitro from Pharbitis as a bioassay system.The PE was prepared from photoperiodically-induced cotyledons (SD-PE). The SD-PE was subjected to the following fractionations: When the SD-PE was extracted with CHCl₃ and then ethyl acetate, the inducing activity was located in the final aqueous fraction. The activity was localized in the diffusate when the aqueous fraction was dialyzed (molecular weight cut off was 10,000). The diffusate was fractionated by ion exchange chromatography, and flower-inducing activity was found in the fraction adsorbed onto anion exchange resin. When the fraction was applied to a Sep-Pak C18 cartridge, the activity eluted with 25% MeOH. As a result of the above fractionation, activity was increased about 30-fold.The nature of the flower-inhibiting activity of the PE taken from cotyledons exposed to continuous-light conditions was examined (CL-PE). The inhibiting activity was decreased as the cotyledons were exposed to longer dark periods; it appeared to be heat-stable. The CL-PE also inhibited flowering in Lemna. The CL-PE was subjected to the following fractionations: When the CL-PE was extracted with CHCl₃ and ethyl acetate, activity was located in the final aqueous fraction. Activity was localized in the diffusate when the aqueous fraction was dialyzed (molecular weight cut off was 10,000). When the diffusate was fractionated by ion exchange chromatography, the activity was found in the flow-through fraction. When the fraction was applied to a hydroxyapatite cartridge, the activity eluted with 25 mM sodium phosphate buffer. When the fraction was re-dialyzed (molecular weight cut off was 1,000), the diffusate contained the activity. As a result of the above fractionation, activity was increased about 10-fold.     

小玉竹的胚胎学研究

小玉竹Polygonatum humile Fisch．ex Maxim．的花药四室．绒毡层腺质型,发育后期出现双核至多核。小孢子四分体左右对称型,偶见四面体型,胞质分裂连续型。成熟花粉具2细胞。子房3室,中轴胎座。胚珠倒生,双珠被,厚珠心,珠孔由内珠被形成。胚囊发育为葱型。受精后,子房壁和外珠被细胞中含有草酸钙针晶。胚发育类似于紫菀型,基细胞有时纵裂形成两个子细胞。胚乳核型。根据实验结果,并结合前人的资料,本文提出了黄精属的胚胎学特征,并在此基础上对黄精族的概念以及属间系统关系进行了探讨。     

乌鸡酶解技术研究

采用猪胰脏作为胰酶的主要来源,对乌鸡的酶解工艺进行了研究。实验结果表明：酶反应的最适温度为４５℃,胰酶混合物的合适用量为乌鸡鲜肉重量的５％,反应１８小时达反应平衡,反应浓度以１ｋｇ鲜乌鸡２０００ｍｌ水为宜,反应过程中维持ｐＨ７．３即可,酶解产物经测定：氨基酸及可溶性短肽总含量高达１６．６１％,其中必须氨基酸含量占氨基酸总含量的５３％。     

枯草芽胞杆菌微生态制剂的研制

采用液体发酵工艺,确定枯草芽胞杆菌的最适发酵条件为:发酵温度30℃,初始pH值7.2,并以1%海藻酸钠和3%明胶组成的混合胶体溶液为囊壁材料,以4%氯化钙作固化剂将枯草芽胞杆菌制成微胶囊剂,稳定性试验结果显示经微胶囊包埋的枯草芽胞杆菌制剂,室温下保存1个月,活菌存活率为98.8%,保存3个月,活菌存活率为50.6%,保存6个月,活菌存活率为15.7%,均高于未经微胶囊化的样品;在4℃冷藏下保存3个月,未经微胶囊化的样品活菌存活率仅为经微胶囊包埋制剂的66.2%。该微胶囊制剂提高了活菌存活率,延长了活菌常温保存期。     

白腐菌紫外诱变选育高产漆酶突变株及其产酶条件研究

以白腐菌为出发菌株,利用紫外线(UV)进行诱变,筛选高产漆酶突变菌株。通过测定致死率绘制出发菌株的致死曲线,采用PDA-RBBR平板变色法进行初筛,ABTS检测酶活对突变株进行摇瓶复筛。结果表明:利用15 w紫外灯在照射距离为30 cm,照射时间为120 s,致死率为72.1%的条件下进行诱变处理,获得一株高产菌株,其酶活提高79.54%,经过5代传代培养,未见酶活下降,具有较好的遗传稳定性,进一步研究了初始pH值,接种量和培养基装液量等对诱变菌株产酶的影响,结果表明在最佳的培养条件pH值6.0,15%的装液量于28℃下,酶活达214.9 U/L。     

四川2种丛生竹木质素和纤维含量的研究

以四川4个地区2种丛生竹(慈竹和硬头黄)为研究对象,对其木质素和综纤维含量进行研究.结果表明,不同地区竹种间综纤维素含量差异显著;除青神地区外,其它几个地区木质素含量差异不大.纬度对木质素积累的作用因竹种而异,对慈竹木质素含量的作用呈极明显的正效应,对硬头黄的作用则呈不明显的正效应,对综纤维含量的影响不大.竹龄影响木质素和综纤维素含量的积累,对慈竹木质素和综纤维的积累呈显著的正效应,但对硬头黄的作用却不明显.研究认为,青神、长宁和绵阳地区的硬头黄以及长宁地区的慈竹是造纸的较好原料.     

Purification and properties of thiosulfate dehydrogenase from Acidithiobacillus thiooxidans JCM7814

A key enzyme of the thiosulfate oxidation pathway in Acidithiobacillus thiooxidans JCM7814 was investigated. As a result of assaying the enzymatic activities of thiosulfate dehydrogenase, rhodanese, and thiosulfate reductase at 5.5 of intracellular pH, the activity of thiosulfate dehydrogenase was measured as the key enzyme. The thiosulfate dehydrogenase of A. thiooxidans JCM7814 was purified using three chromatographies. The purified sample was electrophoretically homogeneous. The molecular mass of the enzyme was 27.9 kDa and it was a monomer. This enzyme had cytochrome c. The optimum pH and temperature of this enzyme were 3.5 and 35 degrees C. The enzyme was stable in the pH range from 5 to 7, and it was stable up to 45 degrees C. The isoelectric point of the enzyme was 8.9. This enzyme reacted with thiosulfate as a substrate. The Km was 0.81 mM.     

内切葡聚糖酶基因在巨大芽孢杆菌中的表达及其酶学性质研究

通过PCR方法将已克隆的内切葡聚糖酶基因(GenBank No. DQ782954)信号肽编码序列去除, 然后与表达载体pHIS1525连接后转化大肠杆菌DH5a, 筛选出阳性转化子DH5 a -pHIS1525-G7并提取质粒进一步转化巨大芽孢杆菌WH320原生质体, 获得基因工程菌WH320-pHIS1525-G7。刚果红染色和SDS-PAGE分析表明该基因在巨大芽孢杆菌中得到了有效表达。基因工程菌经优化培养后, 胞外上清液中的酶活力可达889 U, 是出发菌株(即枯草芽孢杆菌C-36)的11.22倍。酶学性质研究表明: 该酶的最适反应温度与pH值分别为65℃与pH 6.0, 在pH 4.5~10.0范围内50℃保温30 min可保持在最高酶活的80%以上。     

杜仲愈伤组织中次生代谢产物积累动态研究

以杜仲愈伤组织为材料,研究了不同接种量及培养时间、不同来源的愈伤组织及其继代次数对愈伤组织生长和次生代谢产物含量的影响。结果表明,愈伤组织继代培养时接种量在0．35g左右比较合适。愈伤组织的生长曲线大致呈S形,在20d时达到最大值,而总黄酮和绿原酸的含量均在16d时达到最大值。在继代培养中,茎和叶愈伤组织的增长量、绿原酸和总黄酮含量均在第三代达到最大值;下胚轴诱导的愈伤组织的增长量、绿原酸和总黄酮含量均在第四代达到最大值;子叶愈伤组织的增长量和总黄酮含量在第五代达到最大值,绿原酸含量于第四代达到最大值。不同来源的愈伤组织中,叶愈伤组织中绿原酸含最最高,下胚轴愈伤组织中总黄酮含量最高。     

β-葡萄糖苷酶在酿酒酵母表面的表达

应用表面表达技术对来自Trichodermareesei的β-葡萄糖苷酶在酿酒酵母表面的表达及后期性质进行了研究。实验结果表明酵母表面表达酶有活性,该酶的最佳诱导时间为24h,最适温度是70℃,而酶活的最适pH是5．5。使异源表面表达了Bgl1的酵母在以纤维二糖为唯一碳源的培养基中生长,发酵结果表明纤维二糖被明显利用了,但在培养186h后,发酵液中仍残留一定量的纤维二糖。这种技术对纤维素发酵系统中纤维二糖酶活性低的现状有所帮助。     

LIM结构域蛋白FHL1C抑制Notch信号途径的转录激活研究

目的：克隆人表达FHL1C基因以及建立真核表达载体和慢病毒载体。方法：从人的骨骼肌细胞来源的cDNA中扩增并克隆人FHL1C基因的编码区,连接至pMD-18T载体酶切鉴定后测序。序列测定确认后,双酶切pMD18T-FHL1C回收片段,插入真核表达载体,构建真核表达载体pCMV-Myc-FHL1C酶切鉴定正确后,转染Hela细胞及Cos7细胞,用Western Blot检测其在转染细胞中的表达,用双荧光素报告基因系统检测其对Notch信号作用。构建FHL1C-IRES-GFP表达单元,建立慢病毒表达载体plen-ti6/V5-FHL1C-IRES-GFP,包装慢病毒后感染培养的细胞,用免疫荧光显微镜、Western Blot检测分析其在被感染的细胞中的表达。结果：通过PCR方法成功扩增了人FHL1C基因的编码区。通过转染Hela及Cos7细胞,使用Western Blot检测其蛋白水平表达,双荧光报告基因系统分析均能够下调激活的Notch信号。成功构建了FHL1C慢病毒表达载体pLenti6/V5-FHL1C-IRES-GFP,包装慢病毒,把获取的慢病毒感染细胞后通过荧光显微镜证实被感染的细胞绿色荧光蛋白正确表达,Western Blot检测证实其表达。结论：成功建立起FHL1C真核表达载体及慢病毒表达系统,为研究急性T淋巴细胞白血病与Notch信号转导通路之间的关系奠定了基础。     

A case of Amblyomma testudinarium tick bite in a Korean woman

A case of tick bite was found in the inguinal region of a 74-year-old Korean woman. She was attacked by the tick while working in her vegetable garden in the vicinity of mountain located in Suncheon City, the southern coastal area of the Korean Peninsula. On admission she complained of mild discomfort and itching around the bite area. The causative tick was 23 mm long and had slender pedipalps. The scutum was quite ornate and had eyes at the edge. The genital aperture was located anterior to the level of the coxa II. The spiracular plate was comma-shaped and the anus was surrounded posteriorly by the anal groove. The coxa I had subequal 2 spurs; the external one slightly larger. The spur of coxa IV was slightly longer than those of coxae II and III. The tarsus IV had 2 distinct subapical ventral spurs. It was identified as the fully engorged adult female of Amblyomma testudinarium. This is the first human case of Amblyomma bite in Korea.     

小蜡果皮水溶性色素的提取及光敏感性

以去离子水和不同体积分数乙醇从小蜡果皮中浸提红色素,用光谱扫描法检测该红色素的光吸收特性,比较不同浸提时间及不同体积分数乙醇对浸提效果的影响,并用暴露方式进行光敏感性测定。结果显示,小蜡果皮色素在纯水中的溶解性最好,属水溶性色素,延长浸提时间可提高色素的浸出率,但杂质质量分数也随之提高,紫外线对色素的色度影响最大,直接照射可使色素的色度大幅度下降。     

人呼吸道合胞病毒融合蛋白(F)片段原核表达、纯化和鉴定

目的克隆并表达人呼吸道合胞病毒(Human respiratory syncytial virus,HRSV)兰州株的融合蛋白(F)基因片段。方法利用PCR技术扩增HRSV兰州株的融合蛋白基因片段,克隆于原核表达载体pET-42b(+),转化大肠杆菌(Rosetta),经IPTG诱导表达,镍离子亲和层析柱纯化,SDS-PAGE和Western-blot分析重组蛋白的表达及其反应原性。结果 PCR扩增得到951 bp的DNA片段,重组质粒pET42b-F经酶切鉴定和测序分析,表明质粒构建正确。表达的重组蛋白的相对分子质量为68 710,表达的重组蛋白占总菌体蛋白的7%,纯化后蛋白纯度达80%。经Western-blot分析,重组蛋白与抗RSV的单抗呈专一性强阳性反应。结论成功构建了HRSV兰州株F基因片段原核表达载体,并在大肠杆菌Rosetta中获得了表达,表达的重组蛋白具有反应原性和特异性,为HRSV感染引起的疾病血清学诊断以及试剂盒的研发提供了材料。