促性腺激素对猪卵丘细胞-卵母细胞复合体体外减数分裂恢复作用的研究

本实验利用猪卵母细胞体外无血清培养技术,选用猪卵泡液中自然存在的次黄嘌呤（ＨＸ）作为卵母细胞自发成熟的抑制剂,研究了促性腺激素对猪卵丘细胞－卵母细胞复合体（ＣＥＯ）减数分裂恢复的具体作用。ＣＥＯ在含有不同浓度的促性腺激素（ＦＳＨ,ｈＣＧ,ＦＳＨ＋ｈＣＧ）的培养液中培养２４ｈ,观察卵母细胞减数分裂恢复（ＧＶＢＤ）情况。实验结果如下：１．ＦＳＨ（１－５００ＩＵ／Ｌ）能够明显刺激ＣＥＯ克服ＨＸ的抑制作用而恢复减数分裂（Ｐ＜０．０５）,该作用具有剂量依赖性;２．ｈＣＧ（１－５００ＩＵ／Ｌ）对ＣＥＯ减数分裂的恢复无明显作用;３．ｈＣＧ（１０－５００ＩＵ／Ｌ）与ＦＳＨ（１０,１００ＩＵ／Ｌ）无协同作用。上述结果表明,猪ＣＥＯ减数分裂的恢复可能主要依赖于ＦＳＨ的作用,该作用能使猪卵丘细胞产生一种或几种阳性因子,作用于卵母细胞,从而克服ＨＸ的抑制作用而恢复减数分裂。ｈＣＧ无明显作用,可能是因为卵丘细胞上没有ＬＨ受体或ＬＨ受体的数量不足     

谷氨酰胺对体外培养人小肠上皮细胞缺氧复氧损伤的保护作用

应用原代培养人胎小肠上皮细胞（ＩＥＣ）,观察了谷氨酰胺（ＧＬＮ）对缺氧复氧（Ａ／Ｒ）损伤人ＩＥＣ的影响。结果：缺氧６０ｍｉｎ复氧３０ｍｉｎ后,细胞内乳酸脱氢酶（ＬＤＨ）漏出量显著上升,细胞存活率显著下降。预先应用１～５ｍｍｏｌ／ＬＧＬＮ可使Ａ／Ｒ损伤ＩＥＣ细胞存活率升高和细胞内ＬＤＨ漏出量减少,ＧＬＮ作用的最佳剂量为２ｍｍｏｌ／Ｌ。提示ＧＬＮ对人ＩＥＣ具有直接的保护作用,这可能是其整体保护作用机制之一。     

白细胞介素-1β对骨髓基质细胞膜电位及细胞内Ca2+和pH的影响

采用荧光染料 Ｄｉ Ｂ Ａ Ｃ４（３）, Ｆｌｕｏ３／ Ａ Ｍ 和 Ｓ Ｎ Ａ Ｌ Ｆ Ｃａｌｃｅｉｎ／ Ａ Ｍ 分别标记小鼠骨髓基质细胞（ Ｂ Ｍ Ｓ Ｃ）,在激光扫描共聚焦显微镜下直接监测重组人白细胞介素１β（ Ｉ Ｌ１β）刺激后细胞膜电位,细胞内游离 Ｃａ２＋ 浓度和胞浆 ｐ Ｈ 的实时动态变化． 结果发现： Ｉ Ｌ１β加入测定体系后浓度依赖性地引起 Ｂ Ｍ Ｓ Ｃ 膜电位的迅速改变． 低浓度时发生去极化反应,高浓度时发生超极化反应． 非受体方式作用的 Ｉ Ｌ１β肽段 １６３１７１ 对膜电位无影响． Ｉ Ｌ１β不影响细胞内 Ｃａ２＋ 的浓度和胞浆ｐ Ｈ． 研究表明膜电位的变化为 Ｉ Ｌ１ 受体后早期事件,它与细胞内 Ｃａ２＋的浓度和胞浆 ｐ Ｈ 的调节无关．     

促性腺激素对猪卵丘细胞—卵母细胞复合体体外减数分裂？…

本实验利用猪卵母细胞体外无血清培养技术,选用猪卵泡液中自然存在的次黄嘌呤（ＨＸ）作为卵母细胞自发成熟的抑制剂,研究附了中性腺激素对猪卵丘细胞－卵母细胞复体（Ｃ ＥＯ）减数分裂恢复的具体作用。ＣＥＯ在含有不同浓度的促性腺激素（ＦＳＨ,ｈＣＧ,ＦＳＨ＋ｈＣＧ）的培养液中培养２４ｈ,观察卵母细胞减数分裂恢复（ＧＶＢＤ）情况。实验结果如下：１．ＦＳＨ（１－５００ＩＵ／Ｌ）能够明显刺激ＣＥＯ克服ＨＸ的抑制作     

多叶猴耳环──猴耳环属一新种

多叶猴耳环──猴耳环属一新种文和群（广西植物研究所分类研究室,桂林５４１００６）ＰＩＴＨＥＣＥＬＬＯＢＩＵＭＭＵＬＴＩＦＯＬＩＯＬＡＴＵＭ－ＡＮＥＷＳＰＥＣＩＥＳＯＦＰＩＴＨＥＣＥＬＬＯＢＩＵＭＦＲＯＭＧＵＡＮＧＸＩ,ＣＨＩＮＡＷｅｎＨｅｑｕｎ（Ｇｕ...     

高分子量和低分子量尿激酶的分离纯化及动力学性质研究

人尿激酶粗品经苯甲脒亲和柱纯化和Ｐｒｏｔｅｉｎ－ＰａｋＳＰ柱分离后,得到两种分子量的尿激酶（ＵＫ）,即高分子量尿激酶（ＨＵＫ）和低分子量尿激酶（ＬＵＫ）,采用民斯亮蓝法测定蛋白质浓度,纤维蛋白板法测定活力,测得ＨＵＫ比活为２．９×１０＾５ＩＵ／ｍｇ蛋白,ＬＵＫ为３．５１０＾５ＩＵ／ｍｇ蛋白,活力回收为７０％以上,经ＳＤＳ－ＰＡＧＥ鉴定,ＨＵＫ和ＬＵＫ均呈单一条带,分子量分别为５４ｋＤ和３３ｋＤ,Ｈ     

黑龙江铜木蚜蝇雌性的描述

黑龙江铜木蚜蝇雌性的描述ＤＥＳＣＲＩＰＴＩＯＮＯＦＴＨＥＦＥＭＡＬＥＣＨＡＬＣＯＳＹＲＰＨＵＳ（ＸＹＬＯＴＯＭＩＭＡ）ＡＭＵＲＥＮＳＩＳ（ＳＴＡＣＫＥＬＢＥＲＧ）（ＤＩＰＴＥＲＡ：ＳＹＲＰＨＩＤＡＥ）￥ＨＥＪｉｌｏｎｇ;ＣＨＵＸｉｐｉｎｇ（Ｄｅｐａｒ...     

钙离子和蛋白激酶C对大鼠缺血海马脑片谷氨酸堆积的影响

采用大鼠海马脑片体外缺血模型观察钙离子和蛋白激酶Ｃ（ＰＫＣ）对神经元胞外谷氨酸（ＧＬＵ）堆积的影响,结果显示：海马脑片在体外“缺血”１０ｍｉｎ,ＧＬＵ在胞外的浓度增加４倍（从３２±４升高到１１３±１０ｐｍｏｌ／（ｍｉｎ．ｍｇＰｒ）．ｎ＝６）．Ｎ型钙通道拮抗剂蝙蝠葛苏林碱（ＤＳＬ）或无钙培养液均能有效抑制这种浓度的升高（Ｐ＜０．０１）．提示缺血１０ｍｉｎ引发的ＧＬＵ浓度升高是受Ｃａ２＋内流调控的．当脑片在缺血状况下孵育３０ｍｉｎ,ＤＳＬ只部分抑制这种ＧＬＵ堆积,而无钙培养液则无影响,但这额外的ＧＬＵ堆积可被ＰＫＣ抑制剂Ｈ－７完全阻断,而被ＰＫＣ激动剂ＰＤＢ所加强;且不受钙调蛋白抑制剂Ｃａｌｍｄａｚｏｌｉｕｍ和８－溴－ｃＡＭＰ影响．提示缺血３０ｍｉｎ,胞外ＧＬＵ的堆积受钙内流和ＰＫＣ双重调控。     

蛋白激酶C对大鼠缺血海马突触体谷氨酸摄取的调控作用

采用大鼠海马脑片体外缺血模型,观察海马突触体内蛋白激酶Ｃ（ＰＫＣ）活性的变化,以及这种变化对突触体谷氨酸（ＧＬＵ）摄取的影响。结果显示：海马脑片体外“缺血”１０ｍｉｎ,其突触体内ＰＫＣ活性基本不变,而缺血３０ｍｉｎ,突触体内ＰＫＣ活性显著上升（Ｐ＜０．０１,ｎ＝６）;非Ｎ－甲基－Ｄ－天门冬氨酸（ＮＭＤＡ）受体拮抗剂ＤＮＱＸ有效地抑制ＰＫＣ活性的同时,可降低胞外ＧＬＵ的堆积,而ＮＭＤＡ受体阻断剂ＡＰ＿５无作用。进一步实验证明,ＰＫＣ激动剂ＰＤＢ浓度依赖性地抑制突触体对３Ｈ－ＧＬＵ的摄取（ＩＣ５０＝１３１±１０μｍｏｌ／Ｌ）,此抑制作用可由ＰＫＣ抑制剂Ｈ－７（１００μｍｏｌ／Ｌ）抵消。提示脑缺血诱发ＧＬＵ堆积的作用机理可能是：脑缺血引发钙内流导致ＧＬＵ过量释放,ＧＬＵ又通过突触前非ＮＭＤＡ受体激活ＰＫＣ,抑制其自身摄取,正反馈性加重胞外ＧＬＵ的堆积。     

半纤维素的化学结构和生理功能

半纤维素的化学结构和生理功能李雄彪,张金忠（北京大学生物学系北京１００８７１）（南开大学生物系天津３０００７１）ＣＨＥＭＩＣＡＬＳＴＲＵＣＴＵＲＥＡＮＤＰＨＹＳＩＯＬＯＧＩＣＡＬＦＵＮＣＴＩＯＮＯＦＨＥＭＩＣＥＬＬＵＬＯＳＥ￥ＬｉＸｉｏｎｇ－ｂｉａｏ...     

Reagentless amperometric immunosensor for human chorionic gonadotrophin based on direct electrochemistry of horseradish peroxidase

A novel amperometric immunosensor for determination of human serum chorionic gonadotrophin (HCG) was constructed by immobilization of HCG with titania sol-gel on a glassy carbon electrode and the direct electrochemistry of horseradish peroxidase (HRP) labeled to HCG antibody (HRP-anti-HCG). The morphologies of the HCG membrane were characterized to be chemically clean, porous and homogeneous. HRP-anti-HCG was functionally conjugated with the immobilized HCG after incubation in phosphate buffer (PBS) containing HRP-anti-HCG. A direct electron transfer of HRP with a rate constant of 1.35+/-0.40 s(-1) was observed at the HRP-anti-HCG-HCG/titania sol-gel membrane modified electrode in 0.1 M PBS pH 7.0. With a competitive mechanism the differential pulse voltammetric peak current of the immobilized HRP decreased linearly with an increasing HCG concentration from 2.5 to 12.5 mIU/ml in the incubation solution. The HCG immunosensor showed a detection limit of 1.4 mIU/ml, a good accuracy and acceptable precision and reproducibility with an intra-assay CV of 4.7% at 5.0 mIU/ml and an inter-assay precision of 8.1% obtained at 10 mIU/ml. The biosensor displayed a good stability in a storage period of 30 days.     

Progesterone production by dispersed luteal cells of non-pregnant cows: effects of oxytocin and oestradiol

The effects of oxytocin and oestradiol on progesterone production by dispersed luteal cells of non-pregnant cows were studied. In acute incubation (3 h), oxytocin, at a concentration of 800 mIU/ml, significantly inhibited the production of progesterone induced by HCG (10 IU/ml). Suppression of basal progesterone production was evident in some corpora lutea. Lower oxytocin concentrations (4 and 40 mIU/ml) had no effect. At a concentration of 400 mIU/ml, oxytocin may be inhibitory to basal and HCG-induced progesterone production. Oestradiol (1 μkg/ml) had no effect on basal progesterone production but may suppress the production of progesterone induced by HCG. However, incubation with oxytocin (400 mIU/ml) plus oestradiol (1 μg/ml) resulted in a significant inhibition of HCG-induced progesterone production. These data provide evidence for an inhibitory effect of oxytocin on the corpus luteum of non-pregnant cows. Oestradiol may interact with oxytocin to inhibit the bovine corpus luteum function.     

Sensitive time-resolved fluoroimmunoassay for simultaneous detection of serum thyroid-stimulating hormone and total thyroxin with Eu and Sm as labels

Based on a novel cocoating strategy and dissociation enhancement lanthanide fluorescence immunoassay technique, a sensitive time-resolved fluoroimmunoassay (TRFIA) has been developed for simultaneous quantification of human serum thyroid-stimulating hormone (TSH) and thyroxin (T4) in a one-and-the-same assay procedure. The new cocoating strategy for preparing highly active surface anti-TSH and anti-T4 monoclonal antibodies (McAbs) was performed by a three-step protocol. Namely, anti-TSH McAb at high concentration (10 micro g/ml) and extensively biotinylated bovine serum albumin (BSA) at low concentration (0.5 micro g/ml) were coated on microwells by passive adsorption, then streptavidin was captured by the surface BSA-biotin, and finally biotinylated anti-T4 McAb was immobilized by the remnant binding sites of the bound streptavidin. In the present TSH/T4 TRFIA, both sandwich- and competitive-type configurations were involved, and Eu(3+) and Sm(3+) were used as labels for TSH and T4 detection, respectively. The method showed rapid kinetics; the equilibrium was reached within 30min at 37 degrees C due to the use of high concentrations of reaction reagents, rapid agitation, and small reaction volume. The lower limits of detection of the method were 0.028 mIU/L for TSH and 4.1 nmol/L for T4 with 20 micro L of sample volume. The assay ranges for TSH and T4 were 0.21-80.00 mIU/L and 20-300 nmol/L, respectively. The correlation between the TSH/T4 values obtained by the present TSH/T4 TRFIA and those obtained by commercial chemiluminescence immunoassay was satisfactory.     

磁性分离酶联免疫技术检测人类绒毛膜促性腺激素及其临床意义分析

应用磁性分离酶联免疫检测技术（ＭＡＬＡ）对２１０名正常非妊娠妇女的血清进行ＨＣＧ定量测定,平均值３．５４９ｍＩＵ／ｍｌ血清。另外,对１０名不全流产患者、１５名葡萄胎患者及５名早孕误诊患者血清进行ＨＣＧ定量测定,均作出准确诊断。ＨＣＧ定量测定在临床诊断上有重要意义。该法具有灵敏、快速、准确、无污染等特点。是目前临床生殖内分泌激素定量测定的好方法。     

Immunobiological properties of monoclonal antibodies to Mycoplasma hominis antigens

Approaches to obtaining stable mouse hybridomas synthesizing monoclonal antibodies (McAb) to M. hominis key antigens were developed. 4 clones capable of the stable synthesis of McAb of different IgG classes were obtained. Clones A3/2 and A5/D produced antibodies to the thermostable determinant to with a mol. wt. of 80-120 kD, sensitive to sodium periodate and resistant to potassium proteinase. Clone H9/B2 synthesized McAb which interacted with potassium proteinase-sensitive M. hominis thermolabile determinant with a mol. wt. of 80 kD. McAb of clone A3/2, labeled with fluorescein isothiocyanate and horse-radish peroxidase, specifically reacted with M. hominis antigens in the immunofluorescence test and the immunoenzyme assay (EIA). The sensitivity of EIA was 0.25 ng/ml of antigen protein. These data may serve as prerequisites for the development of diagnostic test systems aimed at the detection of M. hominis antigens in different clinical substances.     

Miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion

This paper describes a miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion. The glass fiber membrane was functionally modified with gamma-glycidoxypropyltrimethoxysilane and sodium thiosulfate and was used for separation of protein. Anti-human chorionic gonadotrophin (HCG) immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc), namely, Fc-conjugated IgG (Fc-IgG), was used as a novel analytical reagent. HCG and Fc-IgG complexes were separated from free Fc-IgG based on differences in isoelectric point (pI) using the glass fiber membrane modified with a thiosulfonyl acid functional group. The assay yields a linear relationship between current and HCG concentration in the range of 0-2000 mIU/mL. This simple technique enables the assay of HCG within 2 min. The modified glass fiber membrane was regenerated by occasional elution with malonate buffer (pH 6.0) containing 0.5 M NaCl, to remove free Fc-IgG. Free Fc-IgG recovered in this manner could be reused up to eight times without significant decreases in sensitivity. This miniaturized amperometric flow immunoassay requires only minute quantities of serum and generates highly reproducible results.     

单克隆抗体与抗血清联合应用建立血清CMLC测定方法的研究

单克隆抗体(McAb)和抗血清各有特点。本文提纯人心肌肌球蛋白轻链(CMLC)并制备其单克隆抗体和抗CMLC兔血清(下称抗血清),试建立测定血清CMLC的酶联免疫(ELISA)方法。通过比较,McAb和抗血清联合应用可提高测定方法的灵敏度和特异性;并对各反应试剂的工作浓度进行确定,建立了McAb(1C_(11)-D_7株)为铺底抗体,抗血清为覆盖抗体,碱性磷酸酶标记的羊抗兔IgG为第三抗体的三抗体酶联免疫夹心测定血清CMLC的方法(MP-ELISA)。血清CMLC最小可测浓度为2.5ng/mL。     

The influence of dehydroepiandrosterone on basal and gonadotrophin-stimulated testosterone secretion by Mongolian gerbil interstitial cells incubated or superfused in vitro

1. Testosterone secretion by Mongolian gerbil interstitial cells incubated in the absence of HCG linearly increased with cell concentration (1 x 10(5) cells: 0.6 ng/4 hr, 10 x 10(5) cells: 8.0 ng/4 hr). Addition of 100 mIU HCG resulted in a drastic increase of testosterone secretion which was linear between concentrations of 1 x 10(5) and 4 x 10(5) cells. 2. Compared to HCG-stimulated testosterone release, secretion was significantly higher by cells incubated with 60-100 ng DHEA. 3. During the 4-hr incubation period, 53-69% of added progesterone and 72-88% of added dehydroepiandrosterone (DHEA) were converted to testosterone by cells freshly prepared or stored for 1-3 days at 4 degrees C. On the other hand, prolonged storage at 4 degrees C resulted in a marked decrease of HCG-stimulated testosterone secretion. 4. Testosterone secretion by interstitial cells superfused in vitro increased with the length of HCG (100 mIU/ml) application from 0.08 to 0.22 ng/10(6) cells/min (10 and 60 min, respectively). A much faster and pronounced elevation was found when cells were stimulated with DHEA (200 ng/ml: 0.06-0.80 ng/10(6) cells/min, 0 and 20 min, respectively). 5. After interstitial cells have been stimulated with a DHEA (200 ng/ml) pulse for 30 min and then superfused with medium only for an additional 30 min, testosterone secretion remained significantly elevated and could not be further stimulated by superfusing medium which contained as much as 100 mIU/ml HCG.     

Monoclonal antibodies to Mycoplasma pneumoniae antigens]

Approaches to obtaining stable mouse hybridomas, capable of producing monoclonal antibodies (McAb) to M. pneumoniae key antigens, were developed. As the result of hybridization experiments, 7 clones were obtained; of these, 4 clones stably synthesized IgG McAb. Clones H1/H9 and H9/B2 synthesized antibodies to thermolabile, proteinase-sensitive K protein, produced by cytoplasmic membranes of M. pneumoniae cells. The molecular weight of this protein was found to be 90 kD. McAb of clone H1/H9, labeled with horse-radish peroxidase and fluorescein isothiocyanate, specifically reacted with M. pneumoniae antigens in the immunofluorescence test and the enzyme immunoassay (EIA). The sensitivity of EIA was 0.25 ng/ml of antigen protein. These data are prerequisites for the development of diagnostic test systems for the detection of M. pneumoniae antigens in different biological substances obtained from patients with respiratory pathology.     

抗—HBs国际单位RIA一点法定量测定与计算方法的探讨

在Radio-immuno Assay(RIA)试验测抗－HBs国际单位的简易定量与计算中,采用RIA一点法测定,以分析表达式mIU/ml＝SC（mIU/ml)[exp(0.69315S-NC/SC-NC-1]进行结果计算。此式可以常用对数式表示为mIU/ml=SC(mIU/ml)[lg^-1(0.30103S-NC/SC-NC-1],当（S－NC）在0．3－1．1界线内时,其计算结果的相对误差小于6％,是目前误差最小的简易计算方法。同时还推算求出Holliger公式的阳性对照的最适含量为124．26mIU／ml,Richardson公式的阳性对照为125mIU/ml,中国药品生物制品检定所公式的阳性对照为92．5mIU/ml。