广西黄瓜绿斑驳花叶病毒（CGMMV）黄瓜分离物的RT—PCR检测及CP基因序列分析

利用黄瓜绿斑驳花叶病毒（Cucumber Green Mottle Mosaic Virus,CGMMV）的特异性引物对来自广西一温室栽培的黄瓜病样进行RT—PCR检测,结果扩增得到了与预期大小相符的目的片段（650bp）。序列分析表明,该目的片段包含有CGMMV完整的CP基因序列、部分运动蛋白基因（MP）及3’端非编码区（3＇-UTR）序列,其中CP基因全长486bp,与已报道的CP基因序列同源性为91．2％～99．4％。经系统发育分析,明确该GX-CS分离物与日本、法国、印度等分离物属于CGMMV同一类群,并推测该分离物与广西的葫芦（GX—BG）分离物具有相同的起源关系。     

16S rRNA PCR鉴定嗜酸乳杆菌鸡源分离株

用自行设计的嗜酸乳杆菌16S rRNA的特异性引物和建立的PCR方法对初步鉴定为嗜酸乳杆菌的鸡源分离株进行检测。PCR检测结果表明分离株和嗜酸乳杆菌参考株扩增出大小一致的目的片段,从分子水平对鸡源分离菌进行了鉴定。     

马铃薯S病毒外壳蛋白基因的克隆与原核表达

依据马铃薯S病毒(Potato virus S,PVS)外壳蛋白(CP)基因序列(885bp)设计合成了两对 引物,通过RT-PCR扩增得到长0.8kb的目的片段,将目的片段转入大肠杆菌,酶切鉴定证明 得到了含有目的片段的重组子,测定序列结果与其他PVS分离物CP基因的序列比较,发现其核苷酸同源性达95%左右;构建了含PVS CP基因的融合蛋白原核表达载体,并在大肠杆菌中得到表达,SDS-PAGE测定融合蛋白的分子量为58kD.     

大鼠大脑微血管片段的分离纯化及鉴定

目的:分离大鼠大脑微血管并纯化去除完整的神经细胞,用于克隆血脑屏障上的特异表达基因.方法:采用液相合成法制备粒径200～500 nm的铁氧体磁珠,经两侧颈内动脉插管注入大鼠大脑半球.采用机械分离和酶消化相结合的方法解离脑组织,用筛网滤去组织块和大血管,再在磁场下分选标记磁珠的微血管片段,并从形态学、分子生物学和生物活性角度鉴定获得的脑微血管片段.结果:扫描电镜下没有发现微血管周围存在完整的神经细胞,但在部分区域有胶质的终足包裹.全脑组织微管相关蛋白2a、谷氨酰胺合成酶和CD31的RT-PCR产物均在相应位置出现阳性条带,分离纯化的脑微血管仅CD31阳性.微血管片段内皮细胞摄取的Rh123荧光强度显著低于传代培养的微血管内皮细胞荧光强度.结论:采用本方法可以获得高纯度的、不附带完整神经细胞的脑微血管片段.     

菠菜磷酸丙糖转移蛋白在烟草中正义及反义表达及其对碳水化合物转运的影响

以菠菜（Ｓｐｉｎａｃｉａ ｏｌｅｒａｃｅａ Ｌ．）为材料,取幼叶分离ｍＲＮＡ,反转录合成ｃＤＮＡ,以ｃＤＮＡ第一链为模板,通过ＰＣＲ扩增,获得菠菜磷酸丙糖转移蛋白（Ｔｒｉｏｓｅ ｐｈｏｓｐｈａｔｅ ｔｒａｎｓｌｏｃａｔｏｒ,ＴＰＴ）ｃＤＨＡ目的片段。对其进行序列分析,结果表明,分离的目的片段核苷酸序列与文献报道相比同源率为９９．９％,只不１个碱基发生改变。将得到的菠菜ｔｐｔ ｃＤＮＡ与ＣａＭＶ３５     

染色质免疫沉淀法筛选转录因子Nanog调控的目的基因

目的：利用染色质免疫沉淀法筛选转录因子Nanog调控的目的基因。方法：用甲醛固定胚胎干细胞,利用超声将其染色质随机断裂成0.5～1.0kb的染色质片段,通过免疫沉淀富集与Nanog结合的DNA片段,回复交联后分离纯化DNA片段,最终用凝胶迁移阻滞实验验证Nanog与DNA片段的结合。结果：我们发现fzr为Nanog调控的目的基因,并通过电泳迁移率变动分析证明了二者的结合。结论：Fzr在细胞周期调控中发挥重要作用,这为阐明Nanog的作用机理奠定基础。     

马铃薯S病毒外壳蛋白基因的克隆与原核表达

依据马铃薯S病毒 (PotatovirusS ,PVS)外壳蛋白 (CP)基因序列 (885bp)设计合成了两对引物 ,通过RT PCR扩增得到长 0 .8kb的目的片段 ,将目的片段转入大肠杆菌 ,酶切鉴定证明得到了含有目的片段的重组子 ,测定序列结果与其他PVS分离物CP基因的序列比较 ,发现其核苷酸同源性达 95 %左右 ;构建了含PVSCP基因的融合蛋白原核表达载体 ,并在大肠杆菌中得到表达 ,SDS PAGE测定融合蛋白的分子量为 5 8kD。     

牛副流感病毒3型的分离鉴定

从黑龙江省某牛场患有呼吸道疾病的病牛鼻液中分离到1株病毒, 该病毒能在MDBK细胞上增殖并产生特征性细胞病变, 对分离毒株进行理化特性、RT-PCR鉴定及序列测定与分析, 结果发现该病毒为RNA病毒, 具有血凝性, 对乙醚、氯仿极其敏感, 不耐热, 对酸的抵抗力差; 分离株与GenBank中已发表的病毒株N基因片段的同源性为82.6%~99.1%, 结果表明分离的病毒为牛副流感病毒3型毒株, 命名为BPIV3 DQ1株。根据GenBank上发表的牛副流感病毒3型核衣壳蛋白N基因序列, 设计合成了一对特异性引物, 能扩增704 bp的目的片段, 可以检测出10个 TCID50/100 μL病毒核酸。     

YAC克隆的筛选及鉴定分析技术

人工酵母染色体（ＹＡＣ）技术是人类基因组分析及疾病相关基因的分离、克隆中的关键技术。在基因组ＹＡＣ文库基础上特定目的基因的分离克隆涉及ＹＡＣ克隆的筛选,嵌合体、缺失体和共转染克隆的检测与处理,插入片段的分离及其结构特征的分析,亚克隆的快速构建等等。近年来,有关技术取得了重要进展,已趋于成熟,并正得到广泛应用 。     

YAC克隆的筛选及鉴定分析技术

人工酵母染色体（ＹＡＣ）技术是人类基因组分极及疾病相关基因的分离、克隆中的关键技术。在基因组ＹＡＣ文库基础上特定目的基因的分离克隆涉及ＹＡＣ克隆的筛选,嵌合体、缺失体和共转染克隆的检测与处理,插入片段的分离及及其结构特征的分析,亚克隆的快速构建等等。近年来,有关技术取得了重要进展,已趋于成熟,并正得到了广泛应用。     

一种用于食草动物粪便显微组织分析的临时装片新技术

粪便显微组织分析法是研究食草动物食性的主要方法,其常规装片技术需要使用Hoyer's 装片介质对植物碎片进行封片,而Hoyer's 封片液的粘性易导致植物碎片在装片过程中发生卷曲和重叠,影响植物碎片的识别效果。本文提出的新装片技术采用没有粘性的饱和NaCl 溶液代替Hoyer's 装片介质,结合特定的定量取样方法和装片程序,可以有效地减少植物碎片的卷曲率和重叠率。对比试验显示,新装片技术可使植物碎片卷曲率从10.4% 下降至3.8%,重叠率从25% 下降至8.1% ,说明新装片技术在减少植物碎片卷曲和重叠方面明显优于常规装片方法。     

免疫胶体金技术的应用及展望

免疫胶体金技术是继三大标记技术(荧光素、放射性同位素和酶)后发展起来的固相标记免疫测定技术。免疫胶体金标记技术是以胶体金作为示踪标志物,应用于抗原抗体反应的一种新型免疫标记技术。近年来,该技术在医学、动植物检疫、食品安全监督等各领域得到了日益广泛的应用。从胶体金技术的基本原理、制备方法、标记技术、应用现状及其优点等方面作一简要综述,并对该技术的发展前景作了展望。     

The influence of step-down technique on lower extremity mechanics during curb descent

When stepping down from a curb, individuals typically make initial ground contact with either their rearfoot or forefoot. The purpose of this study was to compare vertical ground reaction forces, lower extremity mechanics, and intra-limb work distribution when individuals adopt a rearfoot technique vs. a forefoot technique, during simulated curb descent. Sixteen subjects stepped down from a platform with both a rearfoot and a forefoot technique. Vertical ground reaction forces and sagittal plane joint kinematics and kinetics were examined for the lead limb during the step-down task. Paired t-tests were used for comparison. Subjects demonstrated greater ankle joint power and negative work, and less hip joint power and negative work, with the forefoot technique vs. the rearfoot technique. Total lower extremity negative work was greater for the forefoot technique vs. the rearfoot technique. The percent contribution to the total negative work was greater for the ankle joint, and less for the hip and knee joints, with the forefoot technique vs. the rearfoot technique. The results of this study may provide insight into how curb descent technique can be modified to alter lower extremity loading.     

An evaluation of techniques for measuring yield turgor in excised Salix leaves

Abstract. The use of three techniques for determining yield turgor in excised Salix leaves was investigated. These were the osmotic-solutions technique, the psychrometer technique, and the pressure-chamber technique. The application of the osmotic-solutions technique to a range of leaf types was discussed and the appropriate corrections for volume changes and the contribution of apoplastic water were detailed. It was concluded that the osmotic-solutions technique is not satisfactory for use with slowly growing and/or very elastic leaves. The psychrometer and pressure-chamber techniques were both simple compared with the osmotic-solutions technique, and gave values for yield turgor in the range of 0·3–0·5 MPa. A disadvantage of the psychrometer technique for field applications is that it requires one psychrometer chamber per sample. The pressure-chamber technique was modified for use as a field technique where multiple sampling could be easily and inexpensively achieved. Particular care was required with this technique to prevent water loss from the leaf during stress relaxation, but simple and effective procedures for doing so were found. The modified pressure-chamber technique described here, is recommended as the preferred technique for measuring the yield turgor of leaves in experiments where many simultaneous estimates of yield turgor are to be made.     

Modified mattress suture technique to correct anterior septal deviation

The purpose of this study was to evaluate the efficacy of a modified mattress suture technique in septal anterior deviation correction and to compare it with the Hinderer technique. This was a randomized, controlled, single-blind clinical trial. Fifty patients with anterior septal deviation were assigned to one of two surgery treatment groups: in group A, 25 patients underwent anterior septoplasty with modified mattress suture technique; and in group B, 25 patients underwent anterior septoplasty with the Hinderer technique. Subjective (oral respiration, epistaxis, rhinorrhea, and nasal obstruction) and objective (anterior rhinomanometry) parameters were analyzed and compared between groups. Better results in obstructive deflections were achieved with the modified mattress suture technique. Unilateral and total nasal resistances improved during the 6-month follow-up in patients who underwent anterior septoplasty with the modified mattress suture technique, and there was a significant difference in the values when compared with patients who underwent anterior septoplasty with rhinoplasty using the Hinderer technique (p = 0.003). The modified mattress suture technique had better subjective and objective results than rhinoplasty with the Hinderer technique and therefore could be considered as an alternative technique in anterior septal surgery.     

A technique using reusable components for hand-infesting cornstalks with European corn borer (Lepidoptera: Crambidae) larvae

Field trials were conducted in 2005 and 2006 to evaluate the use of reusable wire nuts and nonreusable gelatin capsules for hand-infesting cornstalks with European corn borer, Ostrinia nubilalis (Hübner), (Lepidoptera: Crambidae) larvae. The reusable technique, which consists of a modified WingGard plastic wire connector (i.e., wire nut) as a containment device for larvae, was compared over three plant growth stages to a gelatin capsule technique. In 2005 and 2006, the wire nut technique resulted in significantly higher number of wire nuts still intact (i.e., undamaged, with or without a larva) on the stalk at 72 h after infestation compared with the gelatin capsule technique. In addition, the wire nut technique resulted in significantly higher number of tunnels per stalk compared with the gelatin capsule technique at all three corn growth stages during both years. In 2005, the mean +/- SEM number of tunnels per stalk was 0.53 +/- 0.03 in the wire nut technique compared with 0.13 +/- 0.03 tunnels per stalk in the gelatin capsule technique. In 2006, the mean number of tunnels per stalk was 0.45 +/- 0.03 in the wire nut technique compared with 0.08 +/- 0.02 tunnels per stalk in the gelatin capsule technique. In addition, the relative net precision in the wire-nut technique was approximately 2 times higher compared with the gelatin capsule technique.     

Application of wound dressing Molndal technique in clean and potentially contamined postoperative wounds--initial comparative study

Because of a possible delayed wound healing, critical colonization and infection of wounds present a problem for surgeons, particularly in patients with compromised immune system or in case where the wound is heavy contaminated or poorly perfused. Molndal technique of wound dressing has proven to be effective in prevention of infection. In our study we wanted to describe the benefits of the application of Molndal technique wound dressing compared to traditional wound dressing technique at potentially contaminated and clean postoperative wounds. We examined postoperative wound after radical excision of pilonidal sinus and after implantation of partial endoprosthesis in hip fracture. Molndal technique consisted of wound dressing with Aquacel Ag - Hydrofiber. Traditional technique was performed using gauze compresses and hypoallergic adhesives. We analyzed the results of 50 patients after radical excision of pilonidal sinus. 25 patients were treated by Molndal technique and 25 patients by the traditional technique of wound dressing. In the group treated by Molndal technique only 1 (4%) patient has revealed a wound infection, proven by positive microbiological examination and suppuration. In the traditional technique group 4 (16%) patients developed wound infection as inflammation and secretion as a sign of superficial infection. In the other group we analyzed the results of 50 patients after implantation of partial endoprosthesis after hip fracture. 20 patients were treated by Molndal technique and 30 patients by the traditional technique of wound dressing. In the group treated by Molndal technique no patient has revealed a wound infection (0%). In the traditional technique group 4 (13%) patients developed wound infection. All complication in both group were superficial incisional surgical infection (according to HPSC). There was no deep incisional surgical site infection or organ/space surgical site infection. Our results are clearly showing that Molndal technique is effective in preventing the postoperative wound infection.     

A Modified Giemsa C-Banding Technique For Hordeum Species

A Giemsa C-banding technique with a hot 1 N HCI hydrolysis step has been developed for barley chromosomes. This step makes it easy to obtain well separated C-banded chromosomes. To compare this technique with other C-banding techniques, chromosomes of H. vulgare cv. York were stained by both this technique and a modification of the technique of Kimber et al (1976). With respect to centromeric and intercalary bands, both techniques produce a similar banding pattern, but telomeric bands observed by the modified technique of Kimber et al (1976) were not detected by our procedure. This indicates that telomeric heterochromatin may be different chemically and/or structurally from the centromeric and intercalary heterochromatin and its appearance dependent upon the C-banding technique. The procedure described provides a relatively rapid technique for C-banding of barley chromosomes.     

An inertial and magnetic sensor based technique for joint angle measurement

This paper describes the design and evaluation of a miniature kinematic sensor based three dimensional (3D) joint angle measurement technique. The technique uses a combination of rate gyroscope, accelerometer and magnetometer sensor signals. The technique enables 3D inter-segment joint angle measurement and could be of benefit in a variety of applications which require monitoring of joint angles. The technique is not dependent on a fixed reference coordinate system and thus may be suitable for use in a dynamic system such as a moving vehicle. The technique was evaluated by applying it to joint angle measurement of the ankle joint. Experimental results show that accurate measurement of ankle joint angles is achieved by the technique during a variety of lower leg exercises including walking.