绞股蓝总皂甙对单个豚鼠心室肌细胞的钙、钠、钾电流及动作电位的影响

目的研究绞股蓝总皂甙（ＧＰ）对成年豚鼠单个心室肌细胞的动作电位（ＡＰ）、Ｌ－型钙通道电流（ＩＣａ）、快钠通道电流（ＩＮａ）以及内向整流钾通道电流（ＩＫ１）的影响。方法膜片钳全细胞记录。结果５０ｍｇＬ－１ＧＰ能使动作电位时程（ＡＰＤ９０）由给药前的（７０８±２４）ｍｓ缩短到给药后的（３９６±１６）ｍｓ（Ｐ＜０．０１），抑制率５８．１％，动作电位幅值（ＡＰＡ）由给药前的（１２１±７．２）ｍＶ降到给药后的（８６±８．６）ｍＶ（Ｐ＜０．０１），抑制率２４．１％，静息膜电位无明显影响；５～５０ｍｇＬ－１的ＧＰ能浓度依赖性阻滞ＩＣａ和ＩＮａ，但对ＩＫ１无明显影响。结论ＧＰ对缺血心肌的保护作用在于减少“钙超载”和抑制异常兴奋性的产生。     

蝎毒的放射增敏实验研究

目的 探讨蝎毒对放疗的增敏效应及其作用机制。方法 将Ｌｅｗｉｓ肺癌细胞接种于ＮＩＨ小鼠右后肢外侧皮内，当肿瘤平均直径达到６ｍｍ左右时开始实验，观察蝎毒（０．６ｍｇ／ｋｇ）、放疗（４Ｇｙ）及其联合治疗组的肿瘤倍增时间（ＭＤＤＴ）、生存时间（ＡＳＴ）、病理改变和细胞凋亡变化。结果 对照组、蝎毒组、放疗组和蝎毒→放疗组的ＭＤＤＴ分别为（１０．２±０．３４）ｄ、（１１．５±０．３８）ｄ、（１３．７±０．４２）ｄ和（１６．１±０．５３）ｄ，各治疗组均可延长荷瘤小鼠生存期，引起肿瘤组织的坏死，诱发细胞凋亡，联合治疗效果更为突出。结论 蝎毒对荷Ｌｅｗｉｓ肺癌的ＮＩＨ小鼠瘤体不仅有抑制或杀伤作用，而且对放疗有增敏作用。诱发细胞凋亡是其抗肿瘤的主要机制之一。     

鲨骨粉抗吕增效及调节免疫功能的研究

根据祖国医学对鲨鱼药用功效记载,结合现代对鲨鱼软骨的认识,本项研究探讨了鲨鱼软骨抗癌增效及调节免疫功能的作用。结果表明：鲨鱼软骨可明显提高荷瘤小鼠的ＮＫ细胞活性和ＭΦ细胞的吞噬功能。鲨鱼软骨粉与化疗药物（ＣＴＸ、５Ｆｕ、ＡＤＭ）合用,抗肿瘤（Ｓ１８０、Ｌｅｗｉｓ肺癌）的增效率可达２２．８％～４７．９％;与放疗（＾５０Ｃｏ照射）合用,抗肿瘤增效率为１５．２０％～２６．５０％,证实鲨鱼软骨具有较好的扶     

甘草酸三铵盐对兔胸注动脉条收缩反应的影响

甘草酸三铵盐（ＴＡＧＡ）非竞争性对抗去甲肾上腺素（ＮＡ）和氯化钾（ＫＣｌ）对离休兔胸主动脉条的收缩效应，ＰＤ２（ＮＡ）＝６．２４２６±０．３５０８，ＰＤ’２（ＴＡＧＡ）＝１．７８４４±０．３６４５。ＴＡＧＡ同时促进低浓度ＫＣｌ（１０ｍＭｏｌ／Ｌ）收缩动脉条件作用，表现为双向作用。ＴＡＧＡ使ＮＡ所致动脉条收缩发生舒张，且作用持久。在给ＫＣｌ之前，先给ＴＡＧＡ，ＴＡＧＡ抑制ＫＣｌ的收缩作用。     

大鼠后肢热诱导的缓激肽和P物质的释放

用放射免疫法测定缓激肽（ＢＫ）和Ｐ物质（ＳＰ）含量。发现正常大鼠（Ｂ／Ｎ－Ｋｉ）左后肢用４７℃，热水刺激２０ｍｉｎ，后肢足跖皮下灌流液中ＢＫ和ＳＰ含量分别为４３±３４和１１．１±９．７ｆｍｏｌ·ｍｉｎ＾－１，比热刺激前显增加，但在血浆激肽原缺乏大鼠（Ｂ／Ｎ－Ｋａ），ＢＫ和ＳＰ含量分别为１．３±１．０和５．５±３．５ｆｍｏｌ·ｍｉｎ＾－１，明显低于Ｂ／Ｎ－Ｋｉ大鼠。Ｂ／Ｎ－Ｋａ大鼠的Ｅｖａｎｓ蓝漏     

间硝地平和硝苯地平对离体人子宫动脉环的比较性研究

用离体人子宫动脉环来研究间硝地平（ｍ－Ｎｉｆ）和硝苯地平（Ｎｉｆ）的作用：ｍ－Ｎｉｆ和Ｎｉｆ能抑制ＫＣｌ所致的收缩，使其量－效曲线右移，最大反应降低，ＰＤ２’分别为７．８±０．５，７．６±０．７，在无钙液中ＫＣｌ几乎不引起收缩反应，两药抑制ＫＣｌ收缩的效应无明显差别；两药亦能使ＮＥ量－效曲线右移，最大反应降低，ＰＤ２’分别为５．５２±０．１１，４．９８±０．３２，在无钙液中ＮＥ的收缩反应降低为４１％±９．２％，ｍ－Ｎｉｆ抑制ＮＥ收缩作用强于Ｎｉｆ。在离体人子宫动脉环，ｍ－Ｎｉｆ和Ｎｉｆ抑制ＫＣｌ收缩作用远强于抑制ＮＥ收缩作用。     

黄花夹竹桃甙及其与氮芥合用的体内抗肿瘤活性

黄花夹竹桃甙属强心甙类药。１．５ｍｇ·ｋｇ＾－１·ｄ＾－１ｉｐ治疗小鼠腹水瘤Ｈ２２，ＥＡＣ，Ｐ３８８及实体瘤Ｓ１８０，Ｕ１４，Ｌｅｗｉｓ肺癌，仅对实体瘤有效，抑瘤率为４８．７％－５６．７％。ＴＳ与轻度抑瘤作用剂量（０．３，０．５或１．０ｍｇ·ｋｇ＾－１·ｄ＾－１）的氮芥合用，抑瘤活性高于单独给药组，能使腹水瘤小鼠的生存期延长率达到８２．４至＞１２２．１％；对实体瘤的抑瘤率为６５．６％－７２．５％。     

OK-432联合IL-2对C57BL/6小鼠Lewis肺癌抑制的实验研究

目的 研究ＯＫ－４３２与ＩＬ－２联用,抑制近交系小鼠Ｃ５７ＢＬ／６Ｌｅｗｉｓ肺癌的作用。方法 Ｃ５７ＢＬ／６小鼠Ｌｅｗｉｓ肺癌为模型,观察ＯＫ－４３２联合ＩＬ－２对肿瘤发生、生长、转移的影响,及体内对肿瘤细胞的作用。结果 二者联用对肿瘤体积和重量的抑制作用明显增强（Ｐ〈０．０１）,增强抑瘤率（Ｐ〈０．０５）,肺转移灶的数目明显减少。光镜下显示肿瘤灶中出现许多凋亡细胞;肿瘤组织电镜观察,瘤细胞核染色     

牛膝多糖对T淋巴细胞和天然杀伤细胞功能的影响

牛膝多糖（ＡＢＰ）是从中药牛膝根中分离得到的一种有效成分。ＡＢＰ５０－８００ｍｇ·Ｌ－１在体外增强天然杀伤（ＮＫ）细胞活性和促进伴刀豆球蛋白Ａ（ＣｏｎＡ）诱导的肿瘤坏死因子－β（ＴＮＦ－β）产生；但不能提高ＣｏｎＡ诱导的Ｔ淋已细胞增殖反应和白介素２的产生．ＡＢＰ５０及１００ｍｇ·ｋｇ－１ｉｐ明显提高正常小鼠ＮＫ细胞活性和ＴＮＦ─β生成，增强二硝基氟苯诱导的迟发型超敏反应和对抗环磷酰胺对ＮＫ活性的抑制作用。但对ＣｏｎＡ诱导的Ｔ淋巴细胞增殖反应和白介素２的产生无明显影响。表明ＡＢＰ对Ｔ淋巴细胞功能的影响是有选择性的．ＡＢＰ对ＮＫ细胞的杀伤活性的增强作用是明显的．     

二氢埃托啡自动静脉给药对大鼠免疫功能的影响

本研究采用计算机实时控制的定时自动静脉给药法，７２ｈ快速致使大鼠身体依赖性形成。各组戒断症状分值为：ＮＳ组１１．０±ｓ２．９；吗啡组８１．０±ｓ８．６；ＤＨＥ组４１．０±ｓ４．４，表明大鼠已形成明显身体依赖性（Ｐ＜０．００１）。在该模型基础上，利用３Ｈ－ＴｄＲ掺入法测得ＤＨＥ明显抑制大鼠淋巴细胞增殖反应。各组刺激指数为：８０．６９±ｓ１．７６；４９．０４±ｓ１．１０和３０．７７±ｓ２．１４（Ｐ＜０．００１）。利用ＭＴＴ法测ＤＨＥ也能抑制大鼠ＩＬ－２活性，各组Ａ５７０ｎｍ值为：０．３９±ｓ０．０３；０．３１±ｓ０．０４和０．２６±ｓ０．０４（Ｐ＜０．０１）。结果表明ＤＨＥ自动静脉给药明显抑制大鼠免疫功能，且较１０００倍剂量的吗啡作用更强     

3-Methylindole-induced splenotoxicity: functional analysis of immune parameters and lymphocyte phenotyping by flow cytometry.

In this laboratory, 3-methylindole (3-MI), a pneumotoxic metabolite of L-tryptophan that forms in the digestive tract of humans and ruminants, has been demonstrated to be toxic to rat and mouse splenic cells both in vitro and in vivo. The present studies examine whether the reduction in nucleated splenic cells is associated with alterations in: (1) immune functioning (e.g., B and T cell mitogenic responses to lectins), (2) natural resistance (e.g., natural killer (NK) activity and cytokine release from macrophages (MPs)), or (3) the relative percentages of B and T cells in the remaining cells as determined by flow cytometric phenotyping. A dose-dependent decrease in splenic weight (24-46%) and nucleated cell numbers (54-73%) was observed 24 hr after intraperitoneal (ip) administration of 100-300 mg/kg 3-MI to B6C3F1 mice. At a dose of 300 mg/kg, the blastogenic response of splenic lymphocytes to 1 microgram/ml phytohemagglutinin, a T cell mitogen, was reduced 37 and 64%, and NK activity was reduced 20 and 60%, in rats and mice, respectively. Following exposure to 400 mg/kg 3-MI, interleukin-1 and tumor necrosis factor production by lipopolysaccharide-stimulated rat splenic MPs was decreased 58 and 38%, respectively. Despite the reduction in total nucleated cell number in 3-MI-treated mice, the percentages of splenic B and T cells remained the same. These findings indicate that, in addition to its toxicity to splenic cells, 3-MI can significantly impair the functioning of the remaining viable cells. The potential importance of these functional changes for alterations in host resistance in rodents exposed to 3-MI or other alkylindoles is unknown.     

Ganoderma tsugae mycelium enhances splenic natural killer cell activity and serum interferon production in mice.

Effects of the water-soluble extract of Ganoderma tsugae mycelium (GT), its alcohol-insoluble subfraction (GTI), and its alcohol-soluble subfraction (GTS) on splenic natural killer (NK) cell activity and serum interferon (IFN) production were assessed in mice. Intraperitoneal administration of GT (4-200 mg/kg) or GTI (1-50 mg/kg), but not GTS, augmented the NK cytotoxic activity in a dose-dependent manner in C3H/HeN mice. This augmentation of splenic NK cytolytic activity was not mouse-strain-dependent. The serum IFN titers of mice were also elevated after i.p.-doses of GTI. The GTI-induced serum IFN was reduced by either IFN-(alpha+beta) antiserum or IFN-gamma monoclonal antibody in vitro. The treatment with antiserum neutralizing IFN-(alpha+beta) resulted in a 70% reduction of GTI-induced IFN, while monoclonal antibody against mouse IFN-gamma, moderately neutralized the GTI-induced IFN (50%). These results demonstrated that both the splenic NK activity and serum IFN [IFN-(alpha+beta) and IFN-gamma] titers are elevated by Ganoderma tsugae mycelium extracts in mice.     

Tumor site-dependent differential modulation of systemic immunity in RENCA bearing mice

Renal cell carcinoma (RCC) is a highly metastatic cancer which is known to be immunogenic and responsive to immunotherapies using cytokines. The effect of tumor site (organ microenvironment) on the biologic behavior of murine RCC cell line (RENCA) was examined by observing systemic immunomodulations. Reduction of the spleen size observed in tumor bearing mice correlated with a significant decrease in splenic nucleated cell numbers. In the mice bearing RENCA cell tumors in the renal subcapsule, a significant increase of splenic T cell blastogenesis against Con-A (234% of naive control) was observed with reduction of NK activity (45% of naive control). The changes in T cell blastogenesis and NK activity observed in subcutis (s.c.) inoculated RENCA tumor bearers were less significant. However, macrophage functioning and proportions in the spleen were significantly increased only in s.c. tumor bearers. Data indicated tumor site-dependent differential modulation of systemic immunity in RENCA bearing mice which suggests that immunotherapy for RCC should consider not only the tumor but also the tumor's microenvironment.     

弓形体重组质粒pcDNA_3-SAG1免疫小鼠诱导的细胞免疫应答研究

目的 :用 pc DNA3- SAG1真核表达质粒直接免疫小鼠 ,观察 DNA免疫所诱导的小鼠细胞免疫应答 ,为研制弓形虫疫苗奠定基础。方法 :大量制备重组质粒 pc DAN3- SAG1,然后将其导入 BAL B/ c小鼠体内 ,每隔 2 1d接种一次 ,一共免疫三次。用 MTT方法对脾脏的 NK细胞和其淋巴细胞的转化率进行测定 ;采用免疫荧光法对 CD4+、CD8+ 细胞进行测定。结果 :实验组 NK细胞杀伤率为 :70 .0± 3.6 4,而 pc DNA3及空白对照分别为 :48.5± 6 .0 8和 47.0± 5 .93。实验组 NK细胞活性比对照组明显增高 (P<0 .0 5 ) ,而 pc DNA3质粒及空白对照组差异无显著性 (P>0 .0 5 ) ;对 T淋巴细胞亚群 CD4+、CD8+进行动态分析 ,可见随着感染时间的延长 ,CD8+的数量逐渐上升 ,CD4+ / CD8+的比率逐渐下降 ,实验组与 pc DNA3质粒及空白对照有明显差异 (P<0 .0 5 ) ;Con A刺激小鼠淋巴细胞转化实验 ,能刺激免疫鼠及对照鼠淋巴细胞增生 ,实验组与 pc DNA3质粒及空白对照组差异无显著性 (P>0 .0 5 )。结论 :重组质粒pc DNA3- SAG1免疫 BAL B/ c小鼠可诱导一定的细胞免疫。     

Suppressive effects of 3-methylcholanthrene on the in vitro antitumor activity of naturally cytotoxic cells

Transient suppression of splenic natural killer (NK), natural cytotoxic (NC), and peritoneal macrophage cytotoxicity was observed following a single injection of 3-methylcholanthrene (3-MC) into C3H/HeN mice. Natural killer cell activity was depressed by 30-60% 4-6 d after injection of 1.0 mg 3-MC. Levels of NK reactivity returned to normal 8 d post 3-MC injection, and no suppression of natural killing was seen when tested 6 wk after 3-MC treatment. 3-MC did not affect Propionibacterium acnes augmentation of NK cell activity when tested both 6 d and 6 wk after carcinogen injection. The results indicate that the observed suppression of naturally cytotoxic cells may not be important in allowing 3-MC-induced tumors to grow, since suppression is not long-lasting. Therefore, any effect on tumor growth mediated by a suppression of naturally cytotoxic cells would have to be exerted at the earliest stages of tumor development.     

牡蛎提取物抗肿瘤作用的实验研究

以每天０．２ｍｌ剂量的牡蛎提取物连续１０ｄ胃饲荷ＨＡＣ鼠肝癌的Ｂａｌｂ／ｃ小鼠，发现主因瘤而下降的指标明显回升，包括总Ｔ细胞数，Ｔ辅助细胞百分比，丝裂原诱发的淋巴细胞转化强度，和ＮＫ细胞的杀伤活性。瘤重亦较对照组明显减轻，宿主成活期处长。用相同剂量的牡蛎提取物胃饲长有人结肠癌的裸鼠共１５ｄ，发现肿瘤体积明显减少为对照的４４．６％。     

Immunomodulatory effect of polysaccharides from submerged cultured Cordyceps gunnii

Context: The genus Cordyceps (Clavicipitaceae) is a group of entomopathogenic fungi that is widely used as tonic food or invigorant with broad-spectrum medicinal properties in China. Cordyceps gunnii (Berk.)Berk (C. gunnii), is also well known as the Chinese rare caterpillar fungus and has similar pharmacological activities with Cordyceps sinensis (C. sinensis). Polysaccharides (PS) from various Cordyceps species have demonstrated many interesting biological activities, including antitumor, immunopotentiation, hypoglycemic, and hypocholesterolemic activities.

Objective: To investigate the effect of C. gunnii PS on the immunostimulatory antitumor function and expression of immune related cytokines in normal, immuno-suppressive, and H22-bearing mice, respectively. Methods: C. gunnii PS were extracted with hot water at 80°C for 2 h. Normal, immuno-suppressive, and H22-bearing mice were treated with PS respectively. By detecting the value of macrophage phagocytic index, proliferation of lymphocytes, natural killer (NK) cell activity and expression of related cytokines, interleukin (IL-4), tumor necrosis factor-α (TNF-a) and interferon-γ (IFN-γ), and tumor inhibition index in H22-bearing mice additionally, the effect of PS on immunostimulatory antitumor function and its mechanism were studied.

Results: The total sugar content of the PS was determined to be 95% after purification. PS markedly increased the thymus and spleen indexes, the macrophage phagocytosis, the proliferation of splenic cells, and the level of IFN-γ and TNF-α. In tumor growth inhibition test, PS showed remarkable inhibition effects.

Conclusion: PS from the C. gunnii could enhance nonspecific immunological function, humoral immunity, cellular immunity in mice, and inhibit tumor growth.     

Oral administration of a soluble 1–3, 1–6 β-glucan during prophylactic survivin peptide vaccination diminishes growth of a B cell lymphoma in mice

β-glucans are biological response modifiers with activatory effects on macrophages, dendritic cells (DC), granulocytes and NK cells. In this study, we investigated the effect of a soluble yeast-derived β-(1–3), (1–6)-d-glucan on prophylactic peptide vaccination against the B cell lymphoma A20 in syngeneic Balb/c mice. We found that repeated immunizations with two MHC class-I restricted peptides derived from the tumor antigen survivin combined with oral co-administration of β-glucan could significantly diminish intradermal tumor growth, whereas peptide vaccination alone failed to control tumor growth. β-glucan as single agent induced only a weak but non-significant growth inhibitory effect. To determine whether the tumor inhibitory effect of the combined treatment was associated with the induction of a tumor-specific immune response we quantified splenic DC and macrophages, analyzed the maturation of DC and measured the frequency of peptide-specific CD8⁺ and CD4⁺ T cells. Treated mice showed significantly increased numbers of splenic macrophages and mature DC compared to untreated tumor-bearing mice. After restimulation with both peptides in vitro elevated levels of interferon (IFN)-γ-secreting CD8⁺ T cells were found in two of four tested mice following treatment and one of four mice showed a strong increase of interleukin (IL)-4-secreting CD4⁺ T cells. Our data reveal a beneficial effect of β-(1–3), (1–6)-d-glucan in tumor growth inhibition by tumor-specific peptide vaccination which may rely on a function of the polymeric sugar as immunological adjuvant.     

穴位注射LAK细胞对Balb/c小鼠免疫功能的影响

目的:观察小鼠连续多次穴位注射不同数目LAK细胞对其免疫功能的影响,为临床应用提供理论依据。方法:40只小鼠随机分组,每组10只,设为对照组(生理盐水组),阴性对照组(腹腔注射LAK细胞5×10⁷/ml),低剂量组(LAK细胞5×10⁶/ml)和高剂量组(LAK细胞5×10⁷/ml)。另用10只小鼠制备LAK细胞,采用LAK细胞分别对2组小鼠足三里、肝俞穴位注射,每个穴位0.02ml,1次/日,连续7天,对照组注射生理盐水,注射穴位、实验相同。阴性对照组腹腔注射高剂量LAK细胞,每日一次,每次0.1ml,连续7天。结果:阴性对照组与LAK细胞低剂量组的小鼠免疫功能明显高于对照的生理盐水组,NK细胞杀伤活性、NKCF水平以及IL-2活性均有提高,经统计学处理有显著性差异。LAK高剂量组小鼠免疫功能明显高于阴性对照组与低剂量组,NK细胞杀伤活性、NKCF水平以及IL-2活性均有增高,经统计学处理具有显著的差异。结论:LAK细胞对小鼠足三里、肝俞进行穴位注射能明显提高小鼠的免疫功能，高荆量明显优于阴性对照组与低剂t组。同时表明同剂量的穴位注射较非穴位注射更为优越。