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1.
乳鼠外周神经雪旺氏细胞的体外培养和纯化的研究   总被引:3,自引:0,他引:3  
本文取5~7天SD新生大鼠坐骨神经剪碎,用0.25%胰蛋白酶及0.03%胶原酶在37℃消化45分钟,吹打分散后接种于培养瓶。培养基为含10%胎牛血清的F10。接种后48小时,加入阿糖胞苷(Ara-c,Sigma)10-5mol/L处理,48小时后换每毫升含神经生长因子(NGF)100μg的培养液,刺激雪旺氏细胞(SC)生长5~7天。此时根据成纤维细胞去除的情况,可再次加入Ara-C1~2天或转为无血清培养基一周。这种抗有丝分裂法结合无血清培养法,既能有效去除成纤维细胞,又能减少对SC的损伤,获得的SC纯净度可达98%。  相似文献   

2.
鸡胚原始生殖细胞的分离和鸡鸭嵌和体的制备   总被引:2,自引:0,他引:2  
探索了鸡胚12~17期血液中的原始生殖细胞(PGCs)迁移数量变化规律,将其在液氮中冷冻保存.并以Ficoll密度梯度离心、MiniMACS磁气分离、滤膜三种方法对PGCs进行分离,结果发现12~17期血液中均有PGCs存在,13期达到高峰约47.1±10.5个/μl,在血液中比例为0.0126%.冷冻保存3个月后解冻成活率达80%以上.三种分离方法所得的分离效果分别为95.7%、39.2%、63.0%,纯度为27.5%、8.4%、3.1%.将分离的原始生殖细胞以微注射法转移至14~15期麻鸭胚胎中制备了鸡鸭种间嵌和体,获得8只雏鸭(8/110).以鸡W染色体探针原位杂交法在早期鸭胚性腺中检测到鸡原始生殖细胞,嵌和率达84.2%(16/19).表明鸡原始生殖细胞能够迁移定居到鸭胚性腺中,并有可能增殖分化成有功能的配子.  相似文献   

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应用ELISA和PCR法检测502例乙肝病人血清,401例HBsAg阳性血清中,有114例(28.4%)抗-HCV和HCVRNA双项阳性,25例(6.2%)HCVRNA单项阳性;21例(5.2%)抗-HCV单项阳性。将HBsAg乙肝病人分成HBVDNA,HBeAg阳性组和HBVDNA,HBeAg阴性组。前者抗-HCV阳性率为11.6%~20.5%,HCVRNA阳性率为16.2%~20.5%。后者抗-HCV阳性率为20.2%~55.6%,HCVRNA阳性率为23%~60.3%。结果说明长期携带HBV者和慢性乙肝病人均可重叠HCV感染。HBVDNA阳性组抗-HCV和HCVRNA阳性率明显高于HBVDNA阳性组  相似文献   

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荧光法测定羽毛中的色氨酸含量   总被引:1,自引:0,他引:1  
本文探讨了羽毛中色氨酸的荧光分析方法,将2~5mg的羽毛样品置于聚四氟乙烯管中,用1ml含0.5%可溶性淀粉的5mol/L NaOH为水解液,以纯氮为保护气体,在110℃水解20小时,水解产物用6mol/LHCl中和,然后用Na2HPO4-NaOH的缓冲溶液(pH11)稀释5倍,在激发波长280nm,发射波长362nm测定荧光强度及其浓度。经多次试验,结果较为满意,重现性较好,其相对标准偏差为0.61%~3.00%。样品的回收率为95.3%~102.1%。此法快速、简便,适用于各种羽毛的例行分析。  相似文献   

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泡桐叶片蛋白质提取方法的研究   总被引:7,自引:1,他引:6  
24个提取泡桐叶片蛋白质组合的试验结果表明,用UKC(9.5mol/L尿素,5mmol/LK2CO3,1.25%CTAB,0.5%二硫苏糖醇,2%两性载体电解质pH3.5~10,5%TritonX-100)提取由10%冷(-40℃)三氯醋酸(丙酮配制,内含0.07%的巯基乙醇)处理的泡桐叶片干粉提取出的蛋白质总量和种类最多。这表明该方法可用于泡桐叶片蛋白质的提取。  相似文献   

6.
植物材料表面消毒方法的改进   总被引:24,自引:0,他引:24  
苹果枝条用0.1%HgCI2磁力搅拌消毒15-20min,无菌水磁力搅拌清洗5次,每次5min。污染率和杀死率在10%以下,成功率90%左右,且无褐化。  相似文献   

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本实验比较了不同卵龄的小鼠卵母细胞受酒精人工刺激后的激活率和体外受精率,以探讨卵母细胞激活和受精的机制。向NIH雌鼠腹腔注射孕马血清促性腺激素(PMSG)7.5单位,48小时后注射人绒毛膜促性激素(HCG)7.5单位,于不同时间杀小鼠,取卵母细胞与卵丘细胞的复合体(OCC)。从注射HCG后到取OCC的时间视为卵母细胞的卵龄。将OCC置于含8%酒精的M2中7分钟,再在M16中培养5小时后,用0.3mg/mL的透明质酸酶去卵丘细胞。卵母细胞形成原核或速即卵裂为激活的标志。将OCC加入已获能的精子悬液中,5小时后将从卵丘细胞中释放出来的卵母细胞转移到M16中,次日发生卵裂为卵母细胞体外受精和激活的标志。小鼠卵母细胞卵龄为20h,其激活率为81.6%,速即卵裂率为48.0%;而卵龄进一步增加到24h,激活率和卵裂率转为下降(Table1)。而卵母细胞受精子激活和受精则不同,卵龄为15h,卵母细胞的体外受精率为45.4%;随着卵龄的进一步增加,体外受精率则下降(Table2)。Fig.1显示:新排出的卵母细胞容易被精子激活而受精;卵龄较大的卵母细胞较易被酒精的人工刺激而激活。可能是卵母细胞从成熟到老化过程中,细胞的结  相似文献   

8.
采用HGVNS5特异的2对引物,对两个香港株和一个广东株HGVRNA进行逆转录套式PCR扩增,PCR产物克隆入pUC19,重组质粒转化DH5α和JM109菌株。PCR和酶切法鉴定阳性克隆,双脱氧链末端终止法测定核苷酸序列并进行同源性分析。结果发现核苷酸变异呈散在分布,三株间核苷酸和氨基酸序列同源性分别为93.3%~94%及97%~99.2%,与已报道的中国株(CN)相比,则同源性分别为90%~91.2%和94%~96.3%,与美国株(PNF2161及R10291)相比,为87.1%~89.5%和95.2%~97%,而与西非株(GBVC)相比,则达91.4%~93.8%和97%~97.9%。提示HGVNS5区核苷酸和氨基酸序列相对保守,不同HGV株存在一定的地区差异。  相似文献   

9.
本文报道了理化因素对非OI群霍乱弧菌的生长及产生色素的影响。实验结果表明,25℃~36℃、0%-3%NaCl、pH7.4~9.0;42℃、1%~3%NaCl、pH5.0—10.0为最佳生长环境,适宜生长,并有利于色素的产生。  相似文献   

10.
从超数排卵的14只母兔获得438枚受精卵,卵龄16~22小时.显微操作在带微分干涉和相差的Nikon倒置显微镜下进行.注射针的尖端外也0.5μm,离尖端40和80μm处的外径分别为4.2和6.5μm.注射用外源基因是绵羊生长激素基因与MT-1启动基因藕连的线状DNA溶液(1ng/μl).140枚注射的受精卵和未注射的145枚受精卵(对照),在Ham’sF—10培养液(补充生长因子)中培养(38℃,5%的CO2).结果,培养48小时后,注射组卵裂发育率分别是:未卵裂7.9%(11/140)、卵裂至2~4细胞期11.0%(16/140)、卵裂至8~16细胞期80.7%(113/140).对照组相应的卵裂率分别是4.1%(6/145)、12.4%(18/145)和83.4%(121/145).两组卵裂发育率相近.本实验的显微操作对注射后卵的发育没有产生明显的伤害影响.  相似文献   

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It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.  相似文献   

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正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases  相似文献   

16.
Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.  相似文献   

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Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.  相似文献   

20.
Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.  相似文献   

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