木醋杆菌最佳发酵条件

通过试验筛选出木醋杆菌发酵产生纤维素的最佳碳源为葡萄糖,最佳氮源为酵母膏和蛋白胨,并通过正交试验确定出木醋杆菌发酵的最佳条件是:pH 5.0,温度30 ℃,葡萄糖1.5 g/dL,酵母膏0.5 g/dL,蛋白胨1 g/dL.乙醇、醋酸、乳酸对木醋杆菌生产纤维量都有增效作用,优化后的培养基添加0.4 g/dL醋酸,细菌纤维素产量为3.40 g/L.添加体积分数1%的乙醇,细菌纤维素产量为3.65 g/L.添加0.4 g/dL乳酸,细菌纤维素产量为3.54 g/L.     

以基因重组技术开发木聚糖类半纤维素资源

木聚糖类半纤维素是一类取之不尽而又亟待开发利用的碳水化合物，经生物降解后所产生的木糖和少量其它单糖，可以用作基本碳源生产各种发酵产品，包括有机酸、氨基酸、单细胞蛋白、糖醇、工业酶类、溶剂或燃料醇。有关半纤维素酶及其基因的研究已经积累了很多资料，其中包括木聚糖酶、木糖苷酶、阿拉伯糖苷酶、葡萄糖醛酸酶、木聚糖乙酰酯酶以及它们的基因。通过基因重组技术可以使发酵工程菌获得降解半纤维素的能力，或者把能有效降解半纤维素的微生物构建成发酵工程菌，从而把半纤维素转化为所需求的生物工程产品。     

血管紧张素Ⅱ对骨髓间充质干细胞定向诱导为角质形成细胞调控作用的实验研究

目的：探讨骨髓间充质干细胞（MSCs）分化为角质形成细胞的可能性及在此过程中血管紧张素Ⅱ （AngⅡ）对其的调控作用.方法：抽取Wistar大鼠的骨髓,经全骨髓法分离、纯化MSCs,鉴定后建立MSCs细胞模型,用成胶质细胞诱导培养基诱导其为角质形成细胞,显微镜下观察其形态学变化.以添加了AngⅡ 的成角质形成细胞诱导培养基组与单纯成角质形成细胞诱导培养基组在诱导MSCs 7天、10天后行角蛋白10（CKP10）免疫组织化学染色及流式细胞仪检测,并进行对比观察分析.结果：培养的MSCs具有成脂、成骨分化的能力.MSCs可以成功诱导为角质形成细胞,添加Ang Ⅱ的诱导组与对照诱导组CKP10免疫组化染色均有阳性表达,但添加AngⅡ组阳性细胞数高于对照组.流式细胞仪检测显示CKP10阳性细胞百分率,AngⅡ组为80.62％,对照组（32.46％）,两者相差显著（P＜0.05）.结论：MSCs可以诱导分化为角质形成细胞,ANGⅡ对MSC的成角质形成细胞分化有显著的促进作用,这一作用可能是AngⅡ 促进创面愈合的机制之一.     

草菇半纤维素酶系统的诱导、分布及初步定性

以蔗糖、果胶、羧甲基纤维素或木聚糖为碳源培养草菇 (Volvariellavolvacea) ,对所产生的半纤维素酶进行初步研究发现 ,半纤维素酶系主要是由木聚糖诱导 .半纤维素酶中的木聚糖酶分布在胞外 ,木糖糖苷酶分布在胞内 ,阿拉伯糖苷酶胞内胞外都有分布 .木聚糖酶的最适反应温度为 6 0℃ ,最适反应 pH为 5 .8,在pH 5 .4～ 7.0时比较稳定 ,保温 1h酶活的半衰温度是 5 5℃ .木糖糖苷酶的最适反应温度为 5 5℃ ,最适反应 pH为 6 .6 ,在 pH 6 .6～ 7.4时比较稳定 ,保温 1h酶活的半衰温度是 5 2℃ .阿拉伯糖苷酶的最适反应温度为 6 0℃ ,最适反应pH为 5 .0 ,在 pH 4 .6～ 6 .2时比较稳定 ,保温 1h酶活的半衰温度为 6 0℃ .     

脐血间充质干细胞的分离扩增及向成骨及脂肪细胞的分化

目的探讨新生儿脐血间充质干细胞（mesenchymal stem cells，MSCs）体外分离、纯化、扩增，以及向成骨及脂肪细胞定向诱导分化的方法与条件。方法无菌条件下收集新生儿脐血60～120ml，枸橼酸钠抗凝，以Ficoll—Hypaque淋巴细胞分离液密度梯度法、沉降红细胞后密度梯度法及CD34^＋免疫磁珠负选法分离单个核细胞（mononuclear cells，MNCs）。分离获得的MNCs采用L—DMEM培养基或Mesencult^TM培养基／10％胎牛血清进行MSCs培养传代，获得第3代集落生长细胞作流式细胞仪表面抗原测定，并向成骨及脂肪细胞定向诱导分化，成骨细胞钙沉积经茜素红染色鉴定，脂肪细胞胞浆油滴经油红染色鉴定。结果经沉降红细胞后分离的MNCs，使用Mesencult^TM培养基／10％胎牛血清培养成功率高，第3代可出现明显的集落生长，而另两种方法分离培养的细胞则难以形成集落；集落细胞表面抗原测定表达CD29、CD59、CD71而不表达CD34、CD45及HLA—DR等分子。成骨定向诱导分化的集落细胞经茜素红染色胞浆中出现有大量的钙沉积；成脂肪定向诱导分化的集落细胞油红染色示胞浆充满油滴空泡。结论新生儿脐血中可分离出MSCs，并可在体外进行培养扩增。以甲基纤维素沉降红细胞后密度梯度离心分离的MNCs培养较为有效，集落细胞表达基质细胞表面抗原，能够向成骨细胞及成脂肪细胞定向诱导分化。     

Bacillus subtilis木聚糖酶的纯化及其对小麦麸皮的作用

采用活性聚丙烯酰胺凝胶电泳和均质提取法相结合,从枯草芽孢杆菌(Bacillus subtilis)固体培养基发酵产物中分离得到了两种木聚糖酶. 薄层色谱和高压液相色谱分析结果进一步表明它们具有内切木聚糖酶的活力,分别定义为xyl Ⅰ和 xyl Ⅱ.两种酶具有相同的最适反应温度(50℃)和最适pH值(7.0).另外,还研究了内切木聚糖酶 xyl Ⅱ对小麦麸皮不溶性膳食纤维的作用.纸色谱分析结果表明,酶解产物中含有阿魏酰低聚糖.     

激活态雪旺细胞源性神经营养因子对脐血间充质干细胞分化的影响

目的 观察激活态雪旺细胞( ASCs)分泌神经营养因子对诱导人脐血间充质干细胞(HUCBMSCs)神经方向分化的作用.方法 采用双酶消化组织块法结合机械分离法分离培养ASCs;Ficoll法分离得到人脐血单个核细胞(HCMNCs)并经贴壁培养、分离后获得HUCBMSCs.采用全反式微甲酸+碱性成纤维细胞生长因子(bFGF)+表皮生长因子(EGF)(对照组)、ASCs培养基半量换液法(实验组).应用免疫组织化学、蛋白印迹法( Western blot)半定量、荧光实时聚合酶链反应( Real-time PCR)[神经纤维200 (NF200)、胶质纤维酸性蛋白(GFAP)、髓磷脂碱性蛋白(MBP)]定量分析等方法在诱导培养后1、2、3、4周进行综合评价HUCBMSCs诱导分化,对实验结果进行统计学分析.结果 免疫组织化学染色结果证实HUCBMSCs在ASCs和全反式微甲酸及bFGF、EGF的作用下向神经源性细胞分化.不同诱导时间细胞阳性率、Western blot半定量、Real-time PCR定量分析结果显示差异有统计学意义(P＜0.05),而同一时间内两种诱导方法比较差异无统计学意义(P＞0.05).结论 ASCs可以分泌多种神经营养因子并促进HUCBMSCs向神经源性细胞分化,与传统全反式微甲酸诱导方法比较更简单.     

小鼠致密化前胚胎卵裂球体外培养体系的研究

目的探索改善小鼠致密化前胚胎卵裂球体外发育能力。方法用酶消化与机械法结合分离小鼠2-、4-、8-细胞胚胎卵裂球为单个卵裂球及2/4、4/8多卵裂球胚胎,并将其装入空透明带后于兽用人绒毛膜促性腺激素(hCG)注射后138 h,观察透明带包裹、胚胎密度及胰岛素(Ins)、葡萄糖(Glu)添加对各卵裂球、囊胚形成率及囊胚细胞数目的影响。结果透明带包裹提高了2-细胞胚胎卵裂球来源的单腔囊胚的发育率。25/50μl胚胎密度及培养基添加Ins、Glu能显著提高2-细胞胚胎卵裂球囊胚发育率和囊胚细胞数;且对4-和8-细胞胚胎卵裂球发育具有同样的促进作用。结论采用透明带包裹、卵裂球密集培养、培养基添加Ins、Glu的方法成功建立了一种维持致密化前胚胎卵裂球体外高效发育的培养体系。其中,透明带在卵裂球发育中具维持囊胚正常形态作用,避免了多腔囊胚的形成;卵裂球密集培养和培养基添加Ins、Glu均提高囊胚形成率和(或)囊胚细胞数目。该体系有效改善了致密化前胚胎卵裂球体外发育质量。     

毛乳头细胞培养基血清浓度的探索

目的 研究不同血清浓度的培养基对毛乳头细胞生长速度及细胞状态的影响.方法 改良一步酶消化法分离培养人头皮毛乳头细胞,分别以无血清培养基及10%、15%等不同浓度小牛血清培养基,培养第3代毛乳头细胞,倒置相差显微镜下观察细胞生长状态,细胞消化后,计数并绘制生长曲线.结果 在无血清培养基培养条件下,毛乳头细胞传代后贴壁率较低.贴壁后细胞未见明显增殖;传代后前8 d,15%和20%血清培养基培养的毛乳头细胞生长速度,明显快于无血清培养基组和10%血清培养基组(P<0.05);传代第10天后,15%和20%血清培养基中毛乳头细胞生长速度差异无显著的统计学意义.结论 细胞生长速度与血清浓度密切相关.     

干酪乳杆菌蛋白质双向电泳条件优化及图谱建立

采用超声波法和液氮研磨法提取了干酪乳杆菌菌体蛋白质,通过固相pH梯度等电聚焦双向电泳对蛋白质进行分离,并分别利用硝酸银和考马斯亮蓝方法进行染色.对上述不同方法比较结果显示,超声-三氯乙酸/丙酮提取蛋白质方法简便易行,适用于快速制备蛋白质样品,液氮研磨-三氯乙酸/丙酮沉淀法更适合干酪乳杆菌大规模蛋白质提取及组学检测,并且在制备的过程中不发生明显的蛋白质降解的现象.利用液氮研磨-三氯乙酸/丙酮沉淀法制备的干酪乳杆菌蛋白质样品分离效果良好,可获得重复性好和高分辨率的双向电泳图谱,便于后续的质谱及差异蛋白质组的分析,值得同行及相关研究者参考和借鉴.     

Na+‐Dependent Transport of d‐Xylose by Bovine Intestinal Brush Border Membrane Vesicles (BBMV) is Inhibited by Various Pentoses and Hexoses

To detect whether pentoses and hexoses occurring in rumen bacteria or in hemicellulose ingested with feed and partly released in the small intestine have an affinity for the Na⁺‐dependent glucose transporter of the bovine intestinal brush border membrane (BBM), we investigated whether these monosaccharides inhibit Na⁺‐dependent transport of ¹⁴C‐labelled d ‐xylose across the BBM using brush border membrane vesicles (BBMV) isolated from the mid‐jejunum of cows. We used d ‐xylose as the transport substrate, because it has a low affinity for the Na⁺‐dependent glucose transporter and thus its uptake into BBMV is more efficiently competitively inhibited by other sugars than that of d ‐glucose. d‐ Ribose, d ‐mannose and l ‐rhamnose occurring in rumen bacteria significantly inhibited Na⁺‐dependent uptake of d ‐xylose into BBMV, but their inhibitory effect was less than that of d ‐glucose, d ‐xylose and phlorizin. This also applied to l ‐arabinose (and d ‐arabinose), which is, like d ‐xylose and d ‐galactose, a constituent of hemicellulose, and to 2‐deoxy‐d ‐glucose. Of all monosaccharides tested, only d ‐fructose did not affect Na⁺‐dependent d ‐xylose transport. It is concluded that some pentoses and hexoses occurring in rumen bacteria (d ‐ribose, d ‐mannose and l ‐rhamnose) or hemicellulose (l ‐arabinose and d ‐xylose) have a low affinity for the Na⁺‐dependent glucose transporter of the bovine BBM and may therefore be absorbed from the jejunum when released in the small intestine.     

Migration of Acetylated Hemicellulose from Capillary Hemodialyzer to Blood, Causing Scleritis and/or Iritis

From November 1981 to early March 1982, an outbreak of scleritis and/or iritis occurred among patients treated with a Nipro brand NAC series cellulose acetate capillary dialyzer. The rate of incidence with dialyzers produced in 1982 was significantly higher than that with dialyzers produced in 1981. An extract obtained from the dialyzers caused iritis in rabbits after its infusion into an auricula vein. Glycerol, acetylated carbohydrate (AC) derivatives, urethane derivatives, and polypropyleneglycol were found in the extract. AC derivatives caused iritis in rabbits, whereas they caused hyperemia of the bulbar conjunctiva in dogs. The AC derivatives contained xylose and glucose units in a ratio of 1.6-2.3:1. The amounts of AC derivatives were significantly larger in the extracts from 1982 than from 1981 devices. Moreover, another brand, but the same type, of dialyzer, the Cordis Dow 4000, contained a slight amount of them. These facts show that AC derivatives derived from hemicellulose played a primary role in the outbreak.     

透明质酸类填充剂注射美容并发症及其处理的回顾性分析

目的探讨透明质酸类填充剂注射后出现并发症的种类及原因，总结有效的防治方法。方法回顾分析自2010年1月至2013年12月，因透明质酸类填充剂注射并发症而就诊患者的病史，对并发症种类和发生原因进行分析，并总结相应的治疗措施。结果纳入本研究的患者共266例。其中，感染107例，过量注射68例，移位46例，肉芽肿18例，局部痤疮15例，过敏9例，栓塞3例。均采用注射物取出、透明质酸酶注射和抗感染治疗等措施，效果满意。结论感染、过量注射和注射物移位是透明质酸类填充剂注射后最常见的并发症；严格的术前消毒和掌握注射适应证，能有效减少并发症的发生；切开引流并配合抗生素，能有效治疗注射不当引起的重度感染；注射透明质酸酶，对透明质酸过度注射和移位有较好的治疗效果。     

超声波技术治疗皮肤老化临床效果的初步观察

目的观察超声波技术治疗颜面部皮肤老化的临床疗效。方法采用超声波治疗仪的特定声波强度,对60例颜面部皮肤老化者进行治疗。颜面部皮肤老化的治疗10次为一个疗程,每隔一天治疗一次,观察临床效果。针对每例患者治疗前后的对比照片,并做好记录,进行临床疗效的评估。结果本组患者60例,其中额部皱纹:显效12例,有效36例,无效12例;鱼尾纹:显效15例,有效36例,无效9例;眉间皱纹:显效6例,有效42例,无效12例。无一例出现面部红斑及损伤等不良反应。结论使用超声波皮肤治疗仪能改善面部皮肤松弛与老化,操作简单,安全可靠,患者无痛苦,易于接受,是目前具有临床推广价值的方法。     

微血管减压术治疗三叉神经痛2643例临床分析

目的评价微血管减压术治疗三叉神经痛的临床疗效。方法回顾分析微血管减压术治疗三叉神经痛2643例的手术经验及术中所见。结果2643例中治愈2487例,显效76例,有效48例,无效31例,死亡1例。2136例得到3～240个月随访,有1918例治愈,85例显效,39例有效,30例无效,64例疼痛复发（其中有37例行第2次手术后治愈）。结论异常血管压迫三叉神经入根区,神经根周围蛛网膜增厚粘连是三叉神经痛的病因,行该区粘连的蛛网膜分离后,微血管减压是治疗三叉神经痛的有效方法。     

超声波技术治疗皮肤老化临床效果的初步观察

目的观察超声波技术治疗颜面部皮肤老化的临床疗效。方法采用超声波治疗仪的特定声波强度，对60例颜面部皮肤老化者进行治疗。颜面部皮肤老化的治疗10次为一个疗程，每隔一天治疗一次，观察临床效果。针对每例患者治疗前后的对比照片，并做好记录，进行临床疗效的评估。结果本组患者60例，其中额部皱纹：显效12例，有效36例，无效12例；鱼尾纹：显效15例，有效36例，无效9例；眉间皱纹：显效6例，有效42例，无效12例。无一例出现面部红斑及损伤等不良反应。结论使用超声波皮肤治疗仪能改善面部皮肤松弛与老化，操作简单，安全可靠，患者无痛苦，易于接受，是目前具有临床推广价值的方法。     

The characteristics of the anterior communicating artery aneurysm complex by three-dimensional digital subtraction angiography

To elucidate the three-dimensional structure of the anterior communicating artery complex, we performed three-dimensional digital subtraction angiography in anterior communicating (Acom) aneurysm cases. Eighteen patients (six male and 12 female) with Acom aneurysms were studied. The total number of aneurysms was 18, of which three were unruptured and 15 were ruptured. Aneurysmal sizes and angles between the parent artery and each of the two daughter arteries were measured. Two types were defined, based on the daughter arteries. When the sizes of the two daughter arteries were the same, they were defined as AA’ type, and when different, they were defined as AB type. Furthermore, aneurysms were classified into two types based on neck location. Thus, when the neck was located on the extension of the midline of the parent artery, it was defined as classical neck type, and when it was not, it was defined as deviating neck type. There were 11 cases of AA’ type and seven of AB type. In AA’ type all cases were of the classical neck type, and in AB type three cases were of the classical neck type and three were of the deviating neck type. In the deviating neck type, the necks were deviated to the smaller daughter arteries in all cases.     

妊娠晚期合并水痘对妊娠结局的影响

目的探讨妊娠晚期合并水痘对妊娠结局的影响。 方法收集2011年1月至2014年11月首都医科大学附属北京地坛医院妊娠晚期合并水痘的住院孕妇15例,回顾性分析水痘-带状疱疹病毒对母婴的影响。 结果15例晚期妊娠合并水痘的患者中,孕29～34周患者5例（占33.3%）,均无严重并发症,经积极治疗10～14 d痊愈出院,足月后3例孕妇自然分娩,2例孕妇剖宫产分娩,产妇无不良结局,新生儿阿氏评分、体重均在正常范围,随访新生儿30 d无水痘感染病例。孕37～40周患者10例（占66.7%）,治疗过程中分娩8例,其中4例孕妇剖宫产分娩,4例孕妇自然临产分娩;孕妇感染7 d内分娩者3例,感染7 d以上分娩者5例,其中孕妇感染第5天自然分娩的1例新生儿感染水痘,经积极治疗痊愈出院,其余7例新生儿阿氏评分、体重均在正常范围,随访30 d均无水痘感染;8例产妇治疗7～10 d均痊愈出院;另外2例孕妇水痘痊愈后外院分娩,追访母儿分娩顺利,新生儿随访30 d无水痘感染。孕29～34周和孕37～40周感染水痘的两组孕妇,足月后分娩的新生儿水痘感染率分别为0%和10%,差异具有统计学意义（P = 0.048）。 结论妊娠晚期合并水痘的孕妇应积极给予抗病毒和对症治疗以减轻临床症状、避免出现严重并发症,同时尽量推迟分娩,以减少新生儿水痘的发生。     

多镜联合治疗肝外胆管结石3780例

目的 探讨腹腔镜、胆道镜、十二指肠镜联合治疗肝外胆管结石的方法及疗效.方法 回顾性分析1998年3月至2010年6月兰州大学第一医院收治的3780例肝外胆管结石患者的临床资料,根据患者胆管结石不同位置和采用腹腔镜、胆道镜、十二指肠镜的情况,将患者分为"一镜"、"二镜"、"三镜"治疗组;通过患者治疗后的影像学检查和随访结果,比较3组患者治疗的成功率和并发症的发生率.结果 "一镜"治疗组1425例患者,成功率为89.54%(1276/1425),并发症发生率为6.73%(86/1276);149例治疗失败,其中83例接受"二镜"治疗、66例接受"三镜"治疗."二镜"治疗组1792例患者(含转入83例),成功率为95.93%(1719/1792),并发症发生率为4.07%(70/1719);73例治疗失败(转入"三镜"治疗组)."三镜"治疗组785例(含转入139例)患者,成功率为99.75%(783/785),并发症发生率为0.26%(2/783);2例治疗失败患者选择开腹手术治疗.结论 腹腔镜、胆道镜、十二指肠镜联合应用治疗肝外胆管结石可以优势互补,明显提高结石取尽成功率,降低术后并发症的发生率.

Abstract：

Objective To investigate the efficacy of combined application of laparoscope and multiple endoscopes for the treatment of extra-hepatic bile duct stones.Methods The clinical data of 3780 patients with extra-hepatic bile duct stones who were admitted to the First Hospital of Lanzhou University from March 1998 to June 2010 were retrospectively analyzed.According to the condition of bile duct stones,laparoscope,choledochoscope and duodenoscope were applied separately or combinately.All patients were divided into A,B and C groups.Patients in group A were treated by laparoseopy,choledochoscopy or duodenoscopy;patients in group B were treated by choledochoscopy+duodenoscopy.duodenoscopy+laparoscopy or laparoscopy+choledochoscopy;patients in group C were treated by laparoscopy+duodenoscopy+choledochoscopy.The efficacies of different treatment approaches were analyzed by comparing the results of imaging examination and follow-up.Results The curative rate and complication rate of the group A were 89.54%(1276/1425)and 6.73%(86/1276),respectively.Of the 149 patients in the group A who were failed in the treatment,83 patients were transferred to the group B,and 66 patients were transferred to the group C.The curative rate and complication rate of the group B(including 83 patients transferred from the group A)were 95.93%(1719/1792)and 4.07%(70/1719),respectively,and 73 patients who were failed in the treatment were transferred to the group C.The curative rate and complication rate of the group C(including 139 patients transferred from the group C)were 99.75%(783/785)and 0.26% (2/783),respectively,and 2 patients who were failed in the treatment received open surgery.Conclusion Combined application of laparoscope and endoscopes could raise the success rate of stone clearance and decrease the postoperative complications.