人原发性肝细胞肝癌中丙型肝炎病毒C_（33c）抗原及HBxAg的分布及意义

作者应用抗ＨＣＶＮＳ３区Ｃ３３ｃ抗原２Ｂ６株单克隆抗体和抗ＨＢｘＡｇ多克隆抗体,采用ＡＢＣ法对１０２例人原发性肝细胞肝癌（ＰＨＣ）组织进行了ＨＣＶ及ＨＢＶ抗原定位研究。ＨＣＶＣ３。抗原及ＨＢｘＡｇ在ＰＨＣ中的阳性检出率分别是８１．４％及７４．５％,Ｃ３３ｃ抗原或ＨＢｘＡｇ阳性占所检病例９４．１％,相同病例二者同时阳性为６１．８％。１０２例ＰＨＣ中５０例有癌旁肝组织,其Ｃ３３ｃ抗原和ＨＢｘＡｇ的阳性检出率分别是６２％和９２％。ＨＣＶＣ３３ｃ抗原定位于肝癌细胞的胞浆内,胞核未见阳性信号。Ｃ３３ｃ抗原阳性细胞在ＰＨＣ中呈散在、局灶分布为主,在癌旁肝组织呈弥漫分布为主。本文结果提示ＨＣＶ感染在ＰＨＣ的发生中可能起重要作用。     

10个近交系小鼠的微卫星位点分析及其在遗传监测中应用的探讨（一）

选用不同染色体上的８个微卫星位点,应用ＰＣＲ技术对１０个品系的近交系小鼠进行遗传分析,研究结果表明：在无任何亲缘关系的近交系小鼠品系之间,该８个微卫星位点均具有不同的等位基因。含有不同等位基目的微卫星位点所占的百分比从ＭＳＭ／ＭＳ与Ｃ３Ｈ／ＨｅＪ的１００％到（ＣｘＳ）Ｆ与Ｃ５７ＢＬ／６Ｊ间的２５％,平均５６．７２％,在重组近交系间及重组近交系与亲本之间,此比率从（ＣｘＳ）Ｄ或（ＣｘＳ）Ｍ与ＢＡＬＢ／ＣＨｅＡ间的５０％到（ＣｘＳ）Ｄ或（ＣｘＳ）Ｍ与ＳＴＳ／Ａ,（ＣｘＳ）Ｄ与（ＣｘＳ）Ｍ间的２５％,此８个微卫星位点有可能做为在基因水平上进行近交系小鼠遗传监测的标记。     

测量细胞内吞过程中膜受体流动性的新方法

本文首次提出用ＡＢＣ（ＡｖｉｄｉｎＢｉｏｔｉｎＣｏｍｐｌｅｘ）法标记细胞膜受体,通过ＦＲＡＰ（ＦｌｕｏｒｅｓｃｅｎｃｅＲｅｃｏｖｅｒｙＡｆｔｅｒＰｈｏｔｏｂｌｅａｃｈｉｎｇ）技术,测量细胞内吞过程中受体流动性变化的方法。实验选择巨噬细胞ＣｏｎＡ受体,比较了用ＣｏｎＡ－Ｂｉｏｔｉｎ＋Ａｖｉｄｉｎ－ＦＩＴＣ（ＡＢＣ法）和ＣｏｎＡ－ＦＩＴＣ（直接法）标记的膜表面ＣｏｎＡ受体荧光强度和内吞过程中受体的流动性测量结果。结果显示ＣｏｎＡ－Ｂｉｏｔｉｎ＋Ａｖｉｄｉｎ－ＦＩＴＣ标记的巨噬细胞膜受体的平均荧光强度比用ＣｏｎＡ－ＦＩＴＣ标记的平均荧光强度高大约３倍;ＡＢＣ法标记的受体,测量结果误差小、灵敏度高;ＣｏｎＡ刺激１５ｍｉｎ后,巨噬细胞膜表面ＣｏｎＡ受体的扩散系数和荧光恢复率较静息状态时明显下降。讨论了两种标记方法对测量结果的影响     

从基因工程小C肽人胰岛素原类似物（B－R2－A）制备人胰岛素

以融合蛋白的形式,在Ｅ．ｃｏｌｉ中经温度诱导表达了小Ｃ肽人胰岛素原类似物（Ｂ－Ｒ２－Ａ）．表达的融合蛋白可占细胞总蛋白６８％．经磺酸解,及初步分离Ｓ－磺酸型融合蛋白,再经ＣＮＢｒ裂解后,进行还原重组,ＨＰＬＣ分离纯化等步骤,每升发酵液可得到Ｂ－Ｒ２－Ａ约５０ｍｇ。经酶促转化及ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－２５纯化,得重组人胰岛素约２０ｍｇ,其氨基酸组成与人胰岛素相同,并具有与猪胰岛素相同的生物活性．     

从基因工程小C肽人胰岛素原类似物（B—R2—A）制备人胰岛素

以融合蛋白的形式,在Ｅｃｏｌｉ中经温度诱导表达了小Ｃ肽人胰岛素原来似物（Ｂ－Ｒ２－Ａ）,表达的融合蛋白可占细胞总蛋白６８％。经磺酸解,及初步分离Ｓ－磺酸型融合蛋白,再经ＣＮＢｒ裂解后,进行还原重组,ＨＰＬＣ分离纯化等步骤,每升发酵液可得到Ｂ－Ｒ２－Ａ约５０ｍｇ。经酶促转化及ＤＥＡＥ－Ｓｅｐｈａｄｅｘ－Ａ２５纯化,得重组人胰岛素约２０ｍｇ,基氨基酸组成与人胰岛素相同,并具有与猪胰岛素相同的生物活性。     

鼠伤寒沙门氏菌外膜蛋白诱导BALB／C小鼠免疫保护的实验研究

本实验采用超声破碎、ＴｒｉｔｏｎＸ－１００处理和超速离心技术提取了鼠伤寒沙门氏菌（Ｓａｌｍｏｎｅｌｌａｔｙｈｉｍｕｒｉｕｍ,ＳＴＭ）的外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ,ＯＭＰｓ）,其中脂多糖（ＬＰＳ）的含量约为５％。ＯＭＰｓ经ＳＤＳ—ＰＡＧＥ显示１０余条蛋白带。对ＯＭＰｓ诱发ＢＡＬＢ／Ｃ小鼠产生典型的迟发型变态反应（ＤＴＨ）进行了检测。经腹腔免疫的ＢＡＬＢ／Ｃ小鼠用５００ＬＤ５０鼠伤寒沙门氏菌（５０１１５）攻击,１００％可得到保护;用５００ＬＤ５０伤寒杆菌（Ｅ６８６）攻击     

鼠伤寒沙门氏菌外膜蛋白诱发BALB／C小鼠免疫保护的研究

本文采用超声破碎ＴｒｉｔｏｎＸ—１００和超速离心技术提取了鼠伤寒杆菌（Ｓａｌｍｏｎｅｌｌａｔｙｐｈｉｍｕｒｉｕｍ,ＳＴＭ）的外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ,ＯＭＰｓ）,并发现ＯＭＰｓ中脂多糖ＬＰＳ的含量约为５％,ＯＭＰｓ经ＳＤＳ—ＰＡＧＥ显示１０余条蛋白带。对ＯＭＰｓ诱发ＢＡＬＢ／Ｃ小鼠产生典型迟发型变态反应ＤＴＨ和ＩＬ—２的水平进行了检测。经腹腔免疫的小鼠用５００ＬＤ５０鼠伤寒杆菌（５０１１５）攻击,１００％可得到保护;用５００ＬＤ５０伤寒杆菌（Ｅ６８６）攻击,３３．３％可得到交叉保护。免疫ＢＡＬＢ／Ｃ小鼠的Ｔ淋巴细胞,经尾静脉注射给非免疫小鼠,可使后者得到８５．７％的被动免疫保护,上述结果说明ＯＭＰｓ能诱发ＢＡＬＢ／Ｃ小鼠细胞免疫和保护性免疫,并提示成为分子疫苗的可能性。     

喹诺酮类药物抗乙型肝炎病毒体外实验研究

本文以２．２．１５细胞株为模型,以ＨＢｓＡｇ、ＨＢｅＡｇ、ＨＢＶＤＮＡ、细胞存活率为观察指标,综合评价了喹诺酮类药物吡哌酸（ＰｉｐｅｍｉｄｉｃＡｃｉｄ）、氟哌酸（Ｎｏｒｆｌｏｘａｃｉｎ）、环丙氟哌酸（Ｃｉｐｒｏｆｌｏｓｘａｃｉｎ）、氟嗪酸（Ｏｆｌｏｘａｃｉｎ）体外抗ＨＢＶ效果。结果表明：吡哌酸、氟哌酸、环丙氟哌酸、氟嗪酸对ＨＢｓＡｇ、ＨＢｅＡｇ５０％抑制浓度（ＩＤ＿（５０））分别为１１μｇ／ｍｌ、６４μｇ／ｍｌ、９３μｇ／ｍｌ、１０５μｇ／ｍｌ和１９９μｇ／ｍｌ、１１１μｇ／ｍｌ、２４μｇ／ｍｌ、２１７μｇ／ｍｌ,细胞存活率为５０％时的药物浓度（ＣＤ＿（５０））分别为２１９μｇ／ｍｌ、９０μｇ／ｍｌ、１８１μｇ／ｍｌ、１６９μｇ／ｍｌ,在所选定的用药浓度范围内不同程度抑制培养上清液及细胞内ＨＢＶＤＮＡ及其复制中间体的产生。尤其对超螺旋结构ＤＮＡ（ｓｃＤＮＡ）有不完全抑制作用。     

胸苷激酶基因治疗胃癌的体外实验

将单纯疱疹病毒胸苷激酶基因（ＨＳＶ－ｔｋ）导入恶性肿瘤细胞,随后可应用药物丙氧鸟苷（ｇａｎｃｉｃｌｏｖｉｒ, ＧＣＶ）选择性杀死肿瘤细胞．构建了含胸苷激酶与潮霉素磷酸转移酶（ｈｐｈ）融和基因（ＨｙｔＫ）的真核表达载体ＬＸｐｓｐ－ＨｙｔＫ．以脂质体（ｌｉｐｏｆｅｃｔｉｎ）为介导,将这种质粒与仅含潮霉素Ｂ基因的质粒ＬＸＳＨ 分别转染胃癌细胞系ＢＧＣ－８２３,用６０ Ｕ／ｍ ｌ潮霉素Ｂ进行筛选,得到了可稳定传代的阳性克隆,分别命名为ＢＧＣ－ＨｙｔＫ 和ＢＧＣ－Ｈｙ．三种细胞的生长曲线无明显差别．用不同浓度的ＧＣＶ 分别作用于ＢＧＣ－ＨｙｔＫ, ＢＧＣ－Ｈｙ 及ＢＧＣ－８２３,０．０２～２００ μｇ／ｍ ｌ 的ＧＣＶ 对ＢＧＣ－ＨｙｔＫ 细胞有明显的杀伤作用（ＩＣ５０＝ ０．０２ μｇ／ｍ ｌ）,而对另外两种细胞几乎无毒性作用（ＩＣ５０＞ ２００μｇ／ｍ ｌ）．２０ μｇ／ｍ ｌＧＣＶ 作用９６ ｈ 后,仅存在２０％ 的ＢＧＣ－ＨｙｔＫ 就可使周围的大部分ＨＳＶ－ｔｋ－ 的肿瘤细胞死亡,说明存在较显著的“旁观者效应”     

基因工程干扰素（IFN—α2b）抗柯萨奇和单纯疱疹病毒效应

本文采用微量细胞病变抑制法在Ｗｉｓｈ 及Ｖｅｒｏ 细胞上研究了ＩＦＮ—α２ｂ 抗ＣｏｘＢ３ 型及ＨＳＶ—１ 型和ＨＳＶ—Ⅱ型病毒的作用。结果表明,安达芬和干扰能ＩＨＮα２ｂ 在Ｗｉｓｈ 或Ｖｅｒｏ 细胞上５００ＩＵ／ｍｌ 分别可以抗３ｌｏｇ４ 或１０００ＴＣＩＤ５０ ,５０ＩＵ／ｍｌ 可以抗２ｌｏｇ４ 或１００ ＴＣＩＤ５０ ,５ＩＵ／ｍｌ 可以抗１ｌｏｇ４ 或１０ ＴＣＩＤ５０ 的ＣｏｘＢ３ 型或ＨＳＶ—１ 型和ＨＳＶ—Ⅱ型病毒感染细胞。提示安达芬和干扰能均显示明显的抗病毒效应,且二者无明显差别     

S-methylmethionine reduces cell membrane damage in higher plants exposed to low-temperature stress

S-methylmethionine (SMM), an important intermediate compound in the sulphur metabolism, can be found in various quantities in majority of plants. The experiments were designed to determine the extent to which SMM is able to preserve cell membrane integrity or reduce the degree of membrane damage in the course of low-temperature stress. By measuring electrolyte leakage (EL), it was proved that SMM treatment reduced cell membrane damage, and thus EL, during low-temperature stress in both the leaves and roots of peas, maize, soy beans and eight winter wheat varieties with different levels of frost resistance. Investigations on the interaction between SMM and polyamine biosynthesis revealed that SMM increased the quantities of agmatine (Agm) and putrescine (Put) as well as that of spermidine (Spd), while it had no effect on the quantity of spermine (Spn). Using a specific inhibitor, methylglyoxal-bis-guanyl hydrazone (MGBG), it was proved that the polyamine metabolic pathway starting from methionine played no role in the synthesis of Spd or Spn, so there must be an alternative pathway for the synthesis of SMM-induced polyamines.     

活性氧对巨噬细胞呼吸爆发影响及云芝多糖的保护作用

用化学发光法观察到叔丁基氢过氧化物对培养的小鼠腹腔巨噬细胞呼吸爆发有强烈的抑制作用。云芝多糖经腹腔注射后,能增强巨噬细胞呼吸爆发功能对叔丁基氢过氧化物损伤的抵抗力。云芝多糖处理的巨噬细胞谷胱甘肽过氧化物酶基础活力显著提高,在叔丁基氢过氧化物作用下,云芝多糖处理的巨噬细胞仍有较高的谷胱甘肽过氧化物酶活力。说明巨噬细胞的免疫功能与谷胱甘肽过氧化物酶活力有关,非特异性免疫多糖可提高细胞抗氧化能力,减轻活性氧损伤作用。     

L．B－G．O对小鼠U_（14）宫颈癌细胞DNA合成和超微结构的影响

本实验合并应用枸杞有效成份和大蒜有效成份,作用于Ｕ_１４腹水型宫颈癌小鼠,腹腔注射第四天,发现一般状况改善,取腹水观察,癌细胞破损,ＤＮＡ、ＲＮＡ荧光染色强度减弱,有大量白细胞和巨噬细胞围绕;流式细胞分析Ｇ＿１期细胞堆积,超微结构显示胞质中线粒体肿胀,嵴破坏甚至中空,粗面内质网扩大、脱颗粒,肯定了其作用的效果。此因枸杞有效成分活化巨噬细胞与肿瘤细胞紧密结合而起到溶瘤作用,与大蒜有效成分直接杀伤而作用维持短暂结合起来,可大大提高抗癌作用。     

Elimination of phagocytic cells in the spleen after intravenous injection of liposome-encapsulated dichloromethylene diphosphonate. Ultrastructural aspects of elimination of marginal zone macrophages

It is shown ultrastructurally that intravenously injected and liposome-entrapped dichloromethylene diphosphonate (DMDP) causes marked damage to marginal zone macrophages in the mouse spleen which finally results in the elimination of these cells. Marginal zone lymphocytes are also affected by this treatment, probably as a result of action of the enzymes released by dying macrophages. Marginal zone macrophages largely disappear, 24 h after i.v. injection, leaving an open-meshed reticulum in which a few viable and necrotic lymphocytes are present. The proposed method to eliminate macrophages for some time may be used to study functional aspects of these cells in vivo.     

一氧化氮和肿瘤坏死因子在小鼠免疫性肝损伤中的作用及抗肝炎新药SY－801和SY－640的影响

本研究结果表明：一氧化氮（ＮＯ）在卡介苗（ＢＣＧ）加脂多糖（ＬＰＳ）诱导的免疫性肝损伤中呈现双向作用。来源于吞噬细胞的ＮＯ具有损伤作用,而其它来源的ＮＯ则具有保护作用。肿瘤坏死因子（ＴＮＦ）也参与了ＢＣＧ＋ＬＰＳ诱导的肝损伤。枯否氏细胞通过释放ＮＯ及ＴＮＦ而介导肝损伤。抗肝炎新药ＳＹ－８０１及ＳＹ－６４０的保肝机理与它们升高血浆ＮＯ及降低ＴＮＦ基因表达有关。     

INTRASPECIFIC TRANSFER OF HERBICIDE RESISTANCE IN THE RED MICROALGA PORPHYRIDIUM SP. (RHODOPHYCEAE) VIA PROTOPLAST FUSION

Stable progeny doubly resistant to the herbicides sulfometuron methyl (SMM) and diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] (DCMU) were obtained at a frequency of 2% on fusion of protoplasts derived from mutants of Porphyridium sp. (UTEX 637) that were resistant only to SMM (strain SMR) or DCMU (strain DC-2). In the presence of both herbicides, only the fusion progeny could grow; both parental mutants were inhibited. In the absence of SMM, the activity of acetohydroxy acid synthase (AHAS) in the wild-type strain was similar to that in DC-2, exceeding that of SMR by up to 4.5-fold. AHAS activities of all fusion progeny were lower than those of the wild-type strain and DC-2 but higher than that of SMR. In the presence of SMM, AHAS activities of all tested fusion progeny ranged between those of the two parental mutants. This result indicates that both types of AHAS, the type resistant to SMM and the sensitive type, originating from SMR and DC-2, respectively, were expressed in the fusion progeny. In the presence of DCMU, the photosynthetic activity of SMR was completely inhibited, whereas that of DC-2 was unaffected. The photosynthetic activity of the fusion progeny in the presence of DCMU was slightly lower than that of DC-2. Both the cell volume and the DNA content of the fusion progeny were similar to those of the parents. However, the genetic nature of the fusion products has not yet been elucidated. To the best of our knowledge, this is the first report on transfer of herbicide resistance via protoplast fusion in algae.     

双歧杆菌DNA对巨噬细胞MAPK的影响

目的 探索青春型双歧杆菌的DNA对巨噬细胞丝裂素活化的蛋白激酶（MAPK）活性的影响。方法 以激光共聚焦显微镜定量测定小鼠腹腔巨噬细胞MAPK家系中ERK1／2、JNK和p38的含量。结果 双歧杆菌DNA注射组小鼠腹腔巨噬细胞ERK1／2的平均荧光强度明显高于对照组（P〈0．01）,而JNK和p38的平均荧光强度在2组间则差异无显著性（P〉0．05）。结论 青春型双歧杆菌的DNA能提高巨噬细胞ERK1／2的活性,这可能是其激活巨噬细胞的途径之一。     

Assay of glutathione in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection

Glutathione (GSH) in individual mouse peritoneal macrophages was determined by capillary zone electrophoresis with electrochemical end-column amperometric detection at a gold/mercury amalgam microelectrode. A capillary of 20 microm inner diameter was suitable for determination of GSH in an individual macrophage with a good signal-to-noise ratio. Individual macrophages could be drawn into the capillary with the aid of a inverted microscope. Lysing cells was studied in different buffer solutions. 0.01 mol/liter NaOH was selected to lyse macrophages. In this method, the usual calibration curve of GSH could not be used for the quantification of GSH in individual macrophages. It was found that standard GSH injected after analyzing each cell could be served as external standard. The whole cell injection and the lack of necessity of a derivatization reaction lead to more accurate and precise results. The average amount of GSH in an individual mouse peritoneal macrophage is 5.8 fmol, which is consistent with the literature value.     

双歧杆菌对裸鼠腹腔巨噬细胞产生IL—1及IL—6的影响

给裸小鼠腹腔注射活的青春型双歧杆菌,并以小鼠胸腺细胞增殖法及ＥＬＩＳＡ法分别检测了裸鼠腹腔巨噬细胞分泌的ＩＬ１活性及ＩＬ６含量。结果表明：实验组裸鼠腹腔巨噬细胞分泌的ＩＬ１活性以及ＩＬ６含量均显著高于对照组,两者均具有统计学意义（ｐ＜００１）。这提示青春型双歧杆菌可激活巨噬细胞产生ＩＬ１以及ＩＬ６,它们在该菌调节机体免疫反应中可能起一定作用。     

Effect of polysaccharide extracted from Glaciecola polaris on the protection of mouse macrophages from oxidative injury

The effect of the protein-bound polysaccharide extracted from Glaciecola polaris (PSG) was investigated in vitro in the protection of oxidatively-injured mouse macrophages stressed by tert-butyl hydroperoxide (tbOOH) or by oxidatively modified low-density lipoprotein (Ox-LDL). The results showed that PSG treatments to protect the macrophages from oxidative injury was effective and the macrophage colony-stimulating factor (M-CSF) exhibited some similar effects. It was speculated that both M-CSF and PSG could protect macrophages from oxidative injury. The results suggested that the effects of PSG were associated with its capability of inducing M-CSF expression.