棉铃虫被中华卵索线虫感染后血淋巴中还原糖、海藻糖和游离脂肪酸的变化

用3, 5 -二硝基水杨酸法和气相色谱法, 分别对棉铃虫(Helicoverpa armigera) 感染中华卵索线虫(Ovomermis sinensis) 后不同时期血淋巴中还原糖、海藻糖和游离脂肪酸含量变化进行了分析。感染的1-3 d,棉铃虫血淋巴还原糖含量(mg/ ml) (分别为0. 551 ±0 .035、0 .505 ±0. 025、0. 852 ±0. 051) 显著低于对照组(分别为0. 655 ±0 .041、0 .878 ±0. 032、0 .964 ±0 .034); 4-5 d还原糖含量(分别为1. 016 ±0. 051、1. 016 ±0 .042) 显著高于对照组(分别为0. 767 ±0 .041、0. 853±0 .038), 第6 d 还原糖含量(0 .608 ±0. 031) 显著低于对照组(0. 698 ±0 .024)。感染组棉铃虫血淋巴中海藻糖的含量情况与还原糖的不同, 在感染的1-3 d, 棉铃虫血淋巴海藻糖的含量(分别为0 .349 ±0. 029、0.374 ±0. 027、0 .479 ±0. 046) 与对照组(分别为0. 383 ±0 .026、0 .374 ±0 .023、0 .483 ±0. 034) 的基本相同, 没有明显的变化; 4-6 d海藻糖含量(分别为0 .529 ±0. 045、0. 584 ±0 .041、0. 428 ±0. 041) 低于对照组(分别为0. 698 ±0 .042、0. 652 ±0 .032、0 .469 ±0 .038)。无论是感染组还是对照组, 棉铃虫血淋巴中软脂酸(16∶0)、棕榈油酸(16∶1)、硬脂酸(18∶0)、油酸(18∶1)、亚油酸(18∶2     

川西獐牙菜的组织培养与快速繁殖

1　植物名称　川西獐牙菜 (Swertiamussotii)。2　材料类别　根、叶、带节茎段、不带节茎段、顶芽。3　培养条件　诱导愈伤组织及芽分化培养基 :( 1 )MS +NAA 0 .0 4mg·L- 1 (单位下同 ) +IAA 0 .0 4 +KT 0 .0 2～ 0 .0 4 ;( 2 )MS +NAA 0 .0 4 +IAA 0 .0 4 +KT 0 .0 6～ 0 .1 0 ;( 3)MS +NAA 0 .0 4 +IAA 0 .0 4 +KT 0 .1 2～ 0 .1 4;( 4 )MS + 2 ,4 D 0 .2 ;( 5 )MS +2 ,4 D 0 .4。增殖培养基 :( 6)MS +NAA 0 .0 4 +IAA 0 .0 4 +KT 0 .1 0～ 0 .1 2 ;( 7)MS +KT 0 .1 5。生根培养基 :( 8)MS ;( 9) 1 /2MS ;( 1 0 …     

趣蝶莲的离体快速繁殖

1　植物名称　趣蝶莲 (Kalanchoesynsepala)。2　材料类别　不定芽、茎尖、带节茎段。3　培养条件　丛生芽诱导培养基 :( 1 )MS + 6 BA2 .0mg·L- 1 (单位下同 ) +NAA 0 .2 ,( 2 )MS +6 BA 1 .0 +NAA 0 .1 ;增殖培养基 :( 3)MS + 6 BA0 .5 +NAA 0 .0 5 ,( 4 )MS + 6 BA 0 .2 +NAA 0 .0 5 ,( 5 )MS + 6 BA 0 .1 +NAA 0 .0 5 ;生根培养基 :( 6)MS ,( 7)MS +IBA 1 .0 +NAA 0 .2 ,( 8)MS +NAA 0 .5。以上培养基均含 30 g·L- 1 蔗糖、0 .7%琼脂 ,pH 5 .5～5 .8。培养温度 ( 2 8± 2 )℃ ,光照度1 5 0 0～ 2 0 0 0lx ,光照…     

彩叶草的组织培养

1　植物名称　彩叶草 (Coleusblumeivar.ver schaffeltii)。2　材料类别　茎段 (shoots)。3　培养条件　培养基 :( 1 )MS ;( 2 ) 1 /4MS + 6 BA0 .3mg·L- 1 (单位下同 ) ;( 3) 1 /4MS +NAA 0 .0 5 +6 BAl.0 ;( 4 ) 1 /4MS +NAA 0 .1 + 6 BA 2 .0 ;( 5 )1 /2MS + 6 BAl.0 ;( 6) 1 /2MS +NAA 0 .0 5 + 6 BA2 .0 ;( 7) 1 /2MS +NAA 0 .1 + 6 BA 0 .3;( 8)MS +6 BA 2 .0 ;( 9)MS +NAA 0 .0 5 + 6 BA 0 .3;( 1 0 )MS +NAA 0 .1 + 6 BAl.0。上述培养基中加 30 g·L- 1 蔗糖 (sucrose)和 0 .6%的琼脂粉 (agar) ,pH5 .8。…     

罗贝力的组织培养和快速繁殖

1　植物名称　罗贝力 (Lobeliacardinalis)。2　材料类别　带顶芽的嫩茎段 ( youngshoots)。3　培养条件　丛生芽诱导和增殖培养基 :( 1 )MS + 6 BA 0 .5mg·L- 1 (单位下同 ) +NAA 0 .2 ;( 2 )MS + 6 BA 1 .0 +NAA 0 .2 ;( 3)MS + 6 BA2 .0 +IAA 0 .2 ;( 4 )MS + 6 BA 4.0 +NAA 0 .2。壮苗培养基 :( 5 )MS + 6 BA 0 .1 +NAA 0 .1。生根培养基 :( 6)MS +IBA 0 .3+NAA 0 .2。以上培养基均附加 3%白糖 (sugar) ,0 .75 %琼脂 (agar) ,pH 5 .8,培养温度为 ( 2 5± 2 )℃ ,光照 1 2h·d- 1 ,光照度1 0 0 0lx左右。4　生长与分…     

北海道黄杨树的组织培养

1　植物名称　北海道黄杨树 (Euonymusjaponicuscv.Zhuzi) [1 ] 。2　材料类别　成年植株带腋芽的茎段 (caulineseg ment)。3　培养条件　培养基有 :( 1 )MS +NAA 0 .2mg·L- 1 (单位下同 ) + 6 BA 2 .0 ;( 2 )MS +NAA 0 .1 + 6 BA 1 .0 ;( 3)MS +NAA 0 .0 5 + 6 BA 0 .5 ;( 4 )MS +NAA 0 .1 + 6 BA 0 .5 ;( 5 )MS +NAA 0 .1 + 6 BA 1 .0 ;( 6) 1 /2MS +NAA 0 .5 + 6 BA 0 .1。以上培养基含 3%蔗糖、0 .8%琼脂 ,pH 5 .8。培养温度为 ( 2 1±3)℃ ,光照度为 1 5 0 0lx ,光照时间为 1 2h·d- 1 。4　生长与分化情况4.1　无…     

春季与秋季渤海蓝细菌(聚球蓝细菌属)的分布特点

利用荧光显微镜技术研究渤海中蓝细菌 (聚球蓝细菌属 Synechococcus)的分布特点。发现春季 (1 999年 4～ 5月 )蓝细菌生物量较低 (0 .86～ 0 .0 1 mg C/m3,平均 =0 .1 3 ) ,秋季 (1 998年 9～ 1 0月 )较高 (1 6.6～ 0 .3 7mg C/m3,平均 =3 .2 7)。秋季蓝细菌生物量的平均值 (3 .2 7)是春季的 (0 .1 3 ) 2 5倍。虽然不同水层蓝细菌的生物量与水温分布相反 ,但水温仍是影响蓝细菌季节变化差异的主要原因。春秋季蓝细菌生物量垂直变化都是 1 0 m层 >表层 >底层。蓝细菌生物量在浮游植物总生物量中所占比例 (CB/PB) ,秋季是 0 .3 99～ 0 .0 0 3 (平均 0 .0 64) ,春季是 0 .0 47～ 0 .0 0 0 (平均 0 .0 0 3 ) ,两季相差达2 1倍 (平均值 )。小型浮游生物是渤海蓝细菌的主要捕食者 ,蓝细菌在渤海微食物环中有重要作用。     

山椒的组织培养与快速繁殖

1　植物名称　山椒 (Zanthoxylyumpiperitum)。2　材料类别　带腋芽的嫩茎 (tenderstemwithbuds)。3　培养条件　诱导分化培养基 :( 1 )MS + 6 BA2 .0mg·L- 1 (单位下同 ) +NAA 0 .1。增殖培养基 :( 2 )MS + 6 BA 1 .0 +NAA 0 .1 +谷氨酰胺 ( glu tamine) 1 0 0 ;( 3)MS + 6 BA 0 .5 +NAA 0 .0 5 +IBA0 .0 5 +谷氨酰胺 1 0 0。壮苗培养基 :( 4 )MS。上述培养基均含 3%蔗糖 (canesugar)、0 .8%琼脂 (a gar) ,pH 5 .8。生根培养基 :( 5 ) 1 / 4MS(大量元素1 / 4) +IAAl.0 +KT 0 .0 1 ;( 6)l/ 4MS +IBA 2 .0。培养基 ( 5 )和…     

IL-4抑制Lewis肺癌生长和转移的实验研究

探讨 IL -4对 Lewis肺癌生长和转移的抑制作用。将 Lewis肺癌细胞接种于 C57BL /6小鼠皮下。将 2 0只小鼠随机分为两组 ,接种后第 5天起分别给予生理盐水或 IL-4。第 1 8天处死动物 ,测两组肿瘤的平均体积 ,平均瘤重及肺转移灶。 、 组肿瘤的平均体积为 ( 2 .0 0 7± 0 .0 0 6) cm3 ,( 0 .4 0 4± 0 .0 1 0 ) cm3 ;平均瘤重为 ( 2 .83 3± 0 .0 74 ) g、( 0 .586± 0 .0 4 0 ) g,肺转移灶为 ( 5.50 0± 0 .92 6) ,( 3 .3 75± 1 .0 61 )。 组与 I组比差异有显著性 ( P<0 .0 5) ;平均潜伏期为 :8.3 0± 0 .4 9;1 0 .56± 0 .3 6。 IL-4能抑制 Lewis肺癌生长和转移。     

马蹄金的组织培养和植株再生

1　植物名称　马蹄金 (Dichondrarepens)。2　材料类别　下胚轴 (hypocotyl)、子叶 (cotyle don)、叶片 (leaf)和叶柄 (petiole)。3　培养条件　无菌苗培养基 :(1)MS。愈伤组织诱导培养基 :(2 )MS +2 ,4 D 1mg·L- 1(单位下同 ) +NAA 1+6 BA 0 .2 +KT 0 .2 ;(3)MS +2 ,4 D 1+6 BA 0 .2 ;(4 )MS +2 ,4 D 1+KT 0 .2 ;(5 )MS +NAA0 .5 +ZT 0 .2 ;(6 )MS +NAA 0 .5 +6 BA 0 .5 ;(7)MS +2 ,4 D 0 .2 +6 BA 0 .5 ;(8)MS +2 ,4 D 1+6 BA 0 .5 ;(9)MS +2 ,4 D 1+NAA 0 .5 +6 BA 0 .2 +KT 0 .2。愈伤组织分化培…     

带HSV—tk基因的重组腺病毒基因治疗小鼠B16黑色素瘤的实验研究

我父成功开展了携带单纯疹疹Ⅰ型病毒胸腺嘧啶激酶基因（Ｈｅｒｐｅｓｓｉｍｐｌｅｘｖｉｒｕｓｔｈｙ－ｍｎｉｄｉｎｅｋｉｎａｓｅ,ＨＳＶ－ｔｋ）的复制缺陷型重组腺病毒Ａｄ（ＨＳＶ－ｔｋ）结合使用ＧＣＶ治疗Ｃ５７ＢＬ／６小鼠Ｂ１６黑色素瘤的离体及动物试验。     

In vivo antitumor effect of herpes simplex virus thymidine kinase gene therapy in rat hepatocellular carcinoma: feasibility of adenovirus-mediated intra-arterial gene delivery

Transfer of the herpes simplex virus-thymidine kinase gene, followed by the administration of ganciclovir (HSV-tk/GCV), has been a major approach for cancer gene therapy. We investigated the antitumor effect of the HSV-tk/GCV strategy with the rat orthotopic hepatocellular carcinoma (HCC) model and the tumor-selective gene delivery by an adenovirus-mediated gene transfer through the hepatic artery. The complete antitumor effect was demonstrated, after the treatment with GCV in rat HCC established by the implantation of HSV-tk transferred rat HCC cells. The in vivo bystander effect was also observed. The marked infiltration of CD4+ and CD8+ T lymphocytes, macrophages and NK cells were found in the tumor area. After the injection of adenovirus carrying the LacZ gene into the hepatic artery, the selective expression of transgene in the tumor cell was achieved. These findings indicate that the HSV-tk/GCV strategy, using an adenoviral vector, could be a promising avenue for the treatment of hepatocellular carcinoma.     

Transfection of oral cancer cells mediated by transferrin-associated lipoplexes: mechanisms of cell death induced by herpes simplex virus thymidine kinase/ganciclovir therapy

The Herpes Simplex Virus thymidine kinase (HSV-tk) suicide gene/ganciclovir (GCV) approach has been used for the treatment of a variety of cancers. The purpose of the present study was to evaluate the cytotoxic effect of ganciclovir in oral squamous cancer cells, previously transfected with HSV-tk gene delivered by transferrin-associated complexes (Tf-lipoplexes), as well as to investigate the mechanisms involved in the bystander effect and in the process of cell death. The delivery of HSV-tk gene to the oral cancer cells, HSC-3 and SCC-7, mediated by Tf-lipoplexes followed by ganciclovir treatment resulted in essentially 100% cytotoxicity, the observed toxic effect being dependent both on GCV dose and incubation time. Cell death was shown to occur mainly by an apoptotic process. Different experimental approaches demonstrated that the observed cytotoxicity was mainly due to diffusion of the toxic agent into neighbouring, non-transfected cells, via gap junctions. Preliminary in vivo studies in a murine model for oral squamous cell carcinoma have shown a significant inhibition of tumor growth upon injection of Tf-lipoplexes carrying HSV-tk followed by intraperitonal injection of GCV, as compared to controls.     

Transfection of oral cancer cells mediated by transferrin-associated lipoplexes: Mechanisms of cell death induced by herpes simplex virus thymidine kinase/ganciclovir therapy

The Herpes Simplex Virus thymidine kinase (HSV-tk) suicide gene/ganciclovir (GCV) approach has been used for the treatment of a variety of cancers. The purpose of the present study was to evaluate the cytotoxic effect of ganciclovir in oral squamous cancer cells, previously transfected with HSV-tk gene delivered by transferrin-associated complexes (Tf-lipoplexes), as well as to investigate the mechanisms involved in the bystander effect and in the process of cell death. The delivery of HSV-tk gene to the oral cancer cells, HSC-3 and SCC-7, mediated by Tf-lipoplexes followed by ganciclovir treatment resulted in essentially 100% cytotoxicity, the observed toxic effect being dependent both on GCV dose and incubation time. Cell death was shown to occur mainly by an apoptotic process. Different experimental approaches demonstrated that the observed cytotoxicity was mainly due to diffusion of the toxic agent into neighbouring, non-transfected cells, via gap junctions. Preliminary in vivo studies in a murine model for oral squamous cell carcinoma have shown a significant inhibition of tumor growth upon injection of Tf-lipoplexes carrying HSV-tk followed by intraperitonal injection of GCV, as compared to controls.     

HSV-tk/GCV自杀基因系统与重组TNF联合杀伤肿瘤细胞的光学成像检测

目的:应用光学成像方法检测HSV-tk/GCV自杀基因系统与重组改造的人肿瘤坏死因子(recombinanthuman tumor necrosis factor,rmhTNF)的联合使用,对人涎腺腺样囊性癌细胞(adenoid cystic carcinoma,ACC-M)的杀伤效率。方法:实验分为四组,细胞模型对照组、单纯HSV-tk/GCV自杀基因系统治疗组、rmhTNF处理组、自杀基因联合rmhTNF治疗组。将稳定表达胸苷激酶-绿色荧光蛋白(thymidine kinase-green fluorescen protenin,tk-GFP)的ACC-M细胞(ACC-M-tk-GFP)按5 000个/孔接种到96孔板中,24 h后给药治疗;分别于加药前、给药后6 h、给药后24 h进行荧光成像。检测绿色荧光蛋白(green fluorescent protein,GFP)的荧光用于准确显示胸苷激酶(tk)基因表达的肿瘤细胞,检测碘化丙啶(propidium iodide,PI)的荧光用于显示死亡的肿瘤细胞。通过细胞的透射图及其PI和GFP荧光图像,分别计算tk 细胞和tk-细胞的死亡率。结果:HSV-tk/GCV自杀基因系统与rmhTNF联合治疗组的细胞在加药后6 h就已经出现细胞死亡迹象,而单纯HSV-tk/GCV自杀基因系统治疗组的细胞在加更昔洛韦(ganciclovir,GCV)后24 h才出现细胞死亡,单纯rmhTNF治疗组在加药后24 h还没有细胞死亡发生。对于给药后24 h的tk 细胞死亡率,联合治疗组(85.88%)明显高于GCV(38.13%)和rmhTNF单独治疗组(2.97%),并且联合治疗组给药24 h后会引发部分tk-细胞死亡(9.83%)。结论:HSV-tk/GCV自杀基因系统与rmhTNF联合治疗具有显著的协同抗肿瘤效应,能够明显提高对肿瘤细胞的杀伤率;结合光学成像方法,能为更直观方便地检测各组药物分别对tk 细胞和tk-细胞的杀伤率,可为深入研究"旁观者效应"提供参考。     

健脾化瘀中药提高胞嘧啶脱氨酶/单纯疱疹病毒胸苷激酶基因治疗肝细胞癌的实验研究

目的:观察健脾化瘀中药提高胞嘧啶脱氨酶/单纯疱疹病毒胸苷激酶基因治疗肝细胞癌的作用。方法:脂质体lipofectamine将含有双自杀基因的腺病毒载体pAd-CD/TK导人293细胞,收集病毒上清转染人肝癌细胞BEL7402,MTT法测定BEL7402细胞存活率。裸鼠人肝癌模型转染CD/TK双自杀基因后,给予5-FC500mg/kg,GCV 100mg/kg腹腔注射,同时予健脾化瘀中药960复方灌胃。观察肿瘤生长情况。结果:给予前体药物5-FC和GCV后,CD/TK转染细胞被杀死。并表现出较强的旁观者效应。转染细胞比例达到10%即表现出较强的杀伤作用(P<0.01)。健脾化瘀中药960复方具有提高旁观者效应作用,1.67ml/kg和2.5ml/kg960复方含药血清组细胞存活率显著低于对照组(P<0.01)。转染基因组应用5-FC和GCV治疗后,裸鼠肝癌的生长明显受到抑制(P<0.05),抑瘤率39.42%,单用中药组抑瘤率18.04%,中药与CD/TK 5-FC/GCV联合运用组,较单纯CD/5-FC/HSV-tk/GCV对裸鼠肿瘤模型的生长抑制作用更加明显(P<0.05),抑瘤率55.10%。结伦:腺病毒介导CD/TK自杀基因可有效地杀死人肝癌BEL7402细胞,健脾化瘀中药960复方具有显著提高CD/TK双自杀基因对人肝癌细胞的抑杀作用。     

Co-expression of bfl-1 enhances host response in the herpes simplex virus-thymidine kinase/ganciclovir gene therapy system

Anticancer suicide gene therapy using herpes simplex virus-thymidine kinase (HSV-tk) and ganciclovir (GCV) features the unique advantage of being able to elicit brisk host immune response against tumors and the host response reportedly can be potentiated with the co-expression of other appropriate immune- or apoptosis-related genes. We introduced a novel antiapoptotic gene, bfl-1, to test its applicability in the HSV-tk/GCV system. CT-26 murine colon cancer cells transfected with HSV-tk, alone or in combination with bcl-xL or bfl-1, were either grown in vitro or injected into syngeneic mice, followed by GCV administration. The co-expression of bfl-1 was associated with the upregulation of CD95 and CD40 ligand (CD40L) in vitro and with pronounced intratumoral T-lymphocyte infiltration in vivo. These results add to the previous findings that antiapoptotic genes can be used as an adjunctive component in the HSV-tk/GCV system to enhance host immune response against tumors.     

Tanshinone IIA Increases the Bystander Effect of Herpes Simplex Virus Thymidine Kinase/Ganciclovir Gene Therapy via Enhanced Gap Junctional Intercellular Communication

The bystander effect is an intriguing phenomenon by which adjacent cells become sensitized to drug treatment during gene therapy with herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV). This effect is reported to be mediated by gap junctional intercellular communication (GJIC), and therefore, we postulated that upregulation of genes that facilitate GJIC may enhance the HSV-tk/GCV bystander effect. Previous findings have shown Tanshinone IIA (Tan IIA), a chemical substance derived from a Chinese medicine herb, promotes the upregulation of the connexins Cx26 and Cx43 in B16 cells. Because gap junctions are formed by connexins, we hypothesized that Tan IIA might increase GJIC. Our results show that Tan IIA increased GJIC in B16 melanoma cells, leading to more efficient GCV-induced bystander killing in cells stably expressing HSV-tk. Additionally, in vivo experiments demonstrated that tumors in mice with 10% HSV-tk positive B16 cells and 90% wild-type B16 cells became smaller following treatment with the combination of GCV and Tan IIA as compared to GCV or Tan IIA alone. These data demonstrate that Tan IIA can augment the bystander effect of HSV-tk/GCV system through increased gap junction coupling, which adds strength to the promising strategy that develops connexins inducer to potentiate the effects of suicide gene therapy.     

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”