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1.
本文报道一种简单易行的桥粒提取方法。取新鲜牛鼻表皮棘层,以pH2.3含0.5mmol/L苯甲基磺酰氟、0.1mol/L枸椽酸-枸椽酸钠缓冲液浸泡,匀浆,经蔗糖密度梯度离心,得到纯桥粒。电镜证实桥粒结构保存完好,附在桥粒斑上的中丝部分消失。209牛鼻表皮棘层可提取桥粒约100mg。  相似文献   

2.
目的 观察天疱疮抗体对表皮角朊细胞粘合性连接(桥粒及其成分dg2/3)的影响,探讨棘层松解过程中桥粒的动态变化情况。方法采用纯化的天疱疮抗体,建立天疱疮的器官培养模型和棘层松解模型,观察加入天疱疮抗体后不同时相表皮在光学显微镜下和电子显微镜下的结构情况,并用抗dg2/3单克隆抗体进行免疫组织化学染色。结果天疱疮抗体在24小时内仅引起细胞间隙增宽;在48小时时可引起部分桥粒的破坏;72小时时可致胞膜变平滑,桥粒消失。免疫组织化学检测结果表明有桥粒成分的内部化。结论天疱疮抗体对桥粒及其成分的影响是在较晚期开始的,但桥粒的破坏是棘层松解的关键一环。  相似文献   

3.
本文研究结果表明,应用超微结构形态测量其微绒毛、桥粒有助于胸膜恶性间皮瘤和转移性肺腺癌的鉴别。16例胸膜恶性间皮瘤平均微绒毛长宽比值(L/W)为11.76。相反,10例转移性肺腺癌平均微绒毛L/W为5.22,两者之间具有显著差异(P<0.001)。本文测量了16例恶性间皮瘤和10例转移性肺腺癌的桥粒长度,两组平均桥粒长度差异不十分显著,但是,所有16恶性间皮瘤的桥粒长度均有大于1.5μm,相反,10例转移性肺腺癌无1例桥粒长度超过1.5μm,表明恶性间皮瘤具有长的桥粒,而转移性肺腺癌则缺乏长的桥粒。  相似文献   

4.
It has been documented that the binding activity sites of human chorionic gonadotropin (HCG) receptor and kd in human ovarian tumors are different from those in the normal ovary. And some observations suggest that the HCG receptor depends on adenylate cyclase (AC) for its physiological functions. We studied the activity of AC in normal human ovary and ovarian tumors. Five human ovarian specimens and eighteen ovarian tumor specimens were obtained from women patients undergoing gynecological surgery. Ovaries were homogenized and sonicated. The homogenates were centrifuged at 1000 x g for 15 min. After sucrose density gradient ultracentrifugation (78000 x g, 4 h), the membrane fraction was collected from interface between 33% and 37%. The membrane protein approximately 10 micrograms, ATP 1 mmol/L, 3H-ATP 5 x 10(4) cpm, sulphydryl-ethyl alcohol 10 mmol/L, in a final volume of 200 microliters of Tris-HCl buffer (50 mmol/L), pH7.5, containing MgSO4 5 mmol/L, were incubated at 35 degrees C for 10 min. The reaction was stopped by boiling water bath for 3 min. The AC activity (U/mg): of human normal ovary, 67 +/- 8, of cystadenocarcinoma serous, 146 +/- 70; of cystadenocarcinoma mucinous, 289 +/- 83.  相似文献   

5.
目的:分析评价抗表皮棘细胞桥粒抗体、抗桥粒芯糖蛋白1、3抗体(Dsg1、Dsg3)在天疱疮中的诊断价值。方法选择2011年2月-2013年2月在新疆医科大学第一附属医院皮肤科诊治经临床、组织病理及间接免疫荧光确诊为天疱疮的34例患者(试验组),选择同期排除天疱疮的36例患者为对照组。采用间接免疫荧光法(IIF)检测抗表皮棘细胞桥粒抗体,采用酶联免疫吸附法(ELISA)检测 Dsg1、Dsg3;统计各检测指标的敏感性、特异性、阳性预测值(PPV)、阴性预测值(NPV)和诊断符合率。结果试验组和对照组抗表皮棘细胞桥粒抗体的阳性率分别为73.5%(25/34)、2.8%(1/36)。Dsg1抗体阳性率分别为70.6%(24/34)、0.0%(0/36),Dsg3抗体阳性率分别为79.4%(27/34)、5.6%(2/36)。在天疱疮患者中,抗表皮棘细胞桥粒抗体试验的敏感度、特异度、PPV、NPV、诊断符合率分别为73.5%、97.2%、96.2%、79.5%、85.7%,Dsg1分别为70.6%、100.0%、100.0%、78.3%、85.7%,Dsg3分别为79.4%、94.4%、93.1%、82.9%、87.1%。3种指标在天疱疮患者中灵敏度和特异度的差异无统计学意义(P>0.05)。结论同时行血中循环抗体检测与具体靶抗原确认试验检测,能互相补充,减少临床漏诊,提高诊断阳性率。抗天疱疮抗体阳性结果对确诊天疱疮准确性较好,阴性者排除天疱疮的价值较大。  相似文献   

6.
不同成熟期人卵母细胞慢速冷冻的初步研究   总被引:8,自引:0,他引:8  
目的:研究不同成熟期人卵母细胞冻融后超微结构的改变和冷冻保护剂浓度对卵母细胞发育潜能的影响.方法:收集多囊卵巢综合征患者辅助生殖周期获得的卵母细胞进行慢速冷冻.根据冷冻保护剂中蔗糖浓度的不同(0.1,0.2或0.3 mol/L)对不同成熟期的卵母细胞分组,进行发育潜能的实验研究,同时收集部分未冷冻的和冻融后的卵母细胞进行超微结构观察.并将部分冷冻卵母细胞获得的胚胎用于临床患者.结果:0.2 mol/L的蔗糖浓度较0.1 mol/L更有利于各成熟期卵母细胞的冷冻,成熟卵母细胞用0.3 mol/L的蔗糖冷冻效果较好,经体外受精得到的胚胎移植后,获得2例临床妊娠.电镜结果显示,未成熟卵母细胞卵浆内的线粒体和泡状物较成熟卵母细胞少,且分布没有规律;冷冻对成熟卵母细胞超微结构的影响较未成熟卵母细胞大.结论:适当提高冷冻保护剂中蔗糖浓度,有利于各成熟期卵母细胞的冷冻保存,0.3 mol/L浓度的蔗糖冷冻保护剂是成熟卵母细胞冷冻的最佳浓度.慢速冷冻可以引起卵母细胞超微结构的改变.  相似文献   

7.
Background  Streptococcus (S.) oligofermentans is a newly identified bacteria with a yet to be defined mechanism of sucrose metabolism that results in acid production. This study aimed to investigate the biochemical mechanisms of S. oligoferm-entans glucose metaolism.
Methods  The S. oligofermentans LMG21532, Lactobacillus (L.) fermentum 38 and the S. mutans UA140 were used to characterize sucrose metabolism by measuring lactate dehydrogenase (LDH) activity and lactic acid production. Continuous dynamics and high performance capillary electrophoresis were used to determine LDH activity and lactic acid production, respectively, from bacteria collected at 0, 10 and 30 minutes after cultured in 10% sucrose.
Results  These analyses demonstrated that LDH activity of the three bacterial strains examined remained stable but significantly different throughout the sucrose fermentation process. The S. oligofermentans LDH activity ((0.61±0.05) U/mg) was significantly lower than that of L. fermentum ((52.91±8.97) U/mg). In addition, the S. oligofermentans total lactate production ((0.048±0.021) mmol/L) was also significantly lower than that of L. fermentum ((0.958±0.201) mmol/L). Although the S. oligofermentans LDH production was almost double of that produced by S. mutans ((0.32±0.07) U/mg), lactic acid production was approximately one sixth that of S. mutans ((0.296±0.058) mmol/L). Additional tests examining pyruvic acid production (the LDH substrate) demonstrated that lactic acid concentrations correlated with pyruvic acid production. That is, pyruvic acid production by S. oligofermentans was undetectable following sucrose incubation, however, (0.074±0.024) and (0.175±0.098) mmol/L pyruvic acid were produced by S. mutans and L. fermentum, respectively.
Conclusion  S. oligofermentans is incapable of fermenting carbohydrates to produce enough pyruvic acid, which results in reduced lactic acid production.
  相似文献   

8.
选取了优化过程中典型的3种含有不同碳源的培养基配方(其中配方A中含有100g/L葡萄糖,配方B中含100g/L蔗糖,配方C中含100g/L蔗糖和10g/L谷氨酸),进行摇瓶发酵生产腺苷(AR),以测定发酵液的生化参数。通过研究发酵液中的有机酸与腺苷的积累在时序性上的代谢规律,在此基础上提出了通过增强TCA循环为手段以提高目的产物产量的观点,即TCA强化论假说。  相似文献   

9.
人的正常卵巢及巢癌绒毛膜促性腺激素受体的研究   总被引:1,自引:1,他引:0  
Human Chorionic Gonadotropin (HCG) is major physiological luteotropic factors for the human corpus luteum. The observations strongly suggest that the human ovary possesses a gonadotropin receptor in the cell membrane. We studied the HCG receptor in normal human ovary and ovarian tumors. Twenty-three human ovarian specimens and 16 ovarian tumor specimens were obtained from women patients having gynecological surgery. Ovaries were homogenized and sonicated. The homogenates were centrifuged at 2000 g for 15 min. After sucrose density gradient ultracentrifugation (78,000 g, 4 h), two fractions were collected from layer of 33% and interface between 33% and 37%. Thirty micrograms of ovarian protein, 8 ng 125I-HCG and unlabeled HCG in a final volume of 0.5 ml of 0.05 mol/L Tris buffer were incubated at 30 degrees C for 2 h. The results were shown in the table.  相似文献   

10.
铁皮石斛组培快繁技术研究   总被引:1,自引:0,他引:1  
目的 通过研究铁皮石斛种子萌发、小苗增殖和生根的适宜培养基,建立一套铁皮石斛组织培养的快速繁殖体系.方法 以铁皮石斛种子为外植体,研究激素(6-benzylaminopurine和1-naphthleetic acid)不同质量浓度配比、天然添加物(马铃薯汁和香蕉汁)对铁皮石斛种子萌发、小苗增殖和壮苗生根的影响.结果 铁皮石斛种子萌发最适培养基为1/2MS +0.2 mg/L NAA+ 100 g/L马铃薯汁+25 g/L蔗糖汁,小苗增殖最适培养基为1/2MS +0.2 mg/L NAA+ 100 g/L马铃薯汁(或香蕉汁)+25g/L蔗糖汁+0.5g/L活性炭,壮苗与生根最适培养基为1/2MS+ 0.2 mg/L NAA+ 100 g/L香蕉汁+25 g/L蔗糖汁+0.5g/L活性炭.结论 建立了一套铁皮石斛组培快繁体系,可用于大规模生产铁皮石斛组培苗.  相似文献   

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