蝙蝠蛾被孢霉多糖的分离纯化及理化性质

目的提取蝙蝠蛾被孢霉多糖,并对其理化性质进行研究。方法利用水提醇沉法得到粗多糖CPSM,采用硫酸-蒽酮法测定粗多糖含量,考马斯亮蓝法测定蛋白质含量,薄层色谱法检查是否含有单糖。CPSM经纤维素DE-52柱分离得CPSM-1和盐洗脱多糖。两种多糖分别经Sephadex系列凝胶柱分离纯化得到CPSM-1a和CPSM-2,采用光谱法和色谱法测定其单糖组成、分子质量等。结果粗多糖CPSM糖含量为61.4%,蛋白含量为1.8%,不含还原糖。CPSM-1a主要含有甘露糖、半乳糖和葡萄糖,其摩尔比约为5∶2∶3;CPSM-2主要含有甘露糖、半乳糖醛酸、半乳糖、葡萄糖和木糖,其摩尔比约为5∶1∶2∶1∶1。CPSM-1a和CPSM-2重均分子质量分别为8.8×103和7.6×103。结论 CPSM-1a为一水溶性中性多糖,CPSM-2为一水溶性酸性多糖。     

蝙蝠蛾被毛孢菌丝体改善小鼠睡眠的作用

目的了解以人工发酵工艺生产的蝙蝠蛾被毛孢茵丝体对小鼠的改善睡眠作用.方法小鼠每天经口灌胃给予蝙蝠蛾被毛孢茵丝体粉(42,83,250 mg·kg-1·d-1),观察直接睡眠作用,连续给予受试物30 d后进行延长戊巴比妥钠睡眠时间实验、巴比妥钠阈下剂量催眠实验和巴比妥钠睡眠潜伏期实验.结果蝙蝠蛾被毛孢菌丝体剂量达250 mg·kg-1·d-1可显著增强巴比妥钠阈下剂量的催眠作用(P＜0.01),各剂量组(42,83,250 mg·kg-1·d-1)小鼠由巴比妥钠诱导的睡眠潜伏期也明显缩短(P＜0.05,P＜0.01),但未见蝙蝠蛾被毛孢茵丝体的直接睡眠作用和对戊巴比妥钠诱导的小鼠睡眠时间的影响.结论蝙蝠蛾被毛孢菌丝体对小鼠具有一定的改善睡眠作用.     

珠蚌多糖对实验性移植肿瘤及NK细胞活性的作用

目的研究珠蚌多糖对实验性移植小鼠肿瘤以及对自然杀伤细胞（NK细胞）活性的影响。方法采用两种小鼠移植性肿瘤模型，观察珠蚌多糖对在体肿瘤细胞生长的影响；通过脾淋巴细胞与K562肿瘤细胞的共培养，体外检测NK细胞的活性。结果珠蚌多糖200，100mg·kg^-1对小鼠S180肉瘤的抑制率分别达到49．4％和47．1％，对C57BL／6小鼠B16BL6黑色素瘤的抑瘤率分别为39．1％和34．2％；显著提高S180肉瘤小鼠免疫脏器胸腺指数、脾指数；体外10，100μg·mL^-1珠蚌多糖可增强NK细胞对K562细胞的抑制活性。结论珠蚌多糖对实验移植性小鼠肿瘤生长有明显的抑制作用，体外一定剂量范围内可诱导NK细胞活性。     

灵芝多糖/硒化卡拉胶口服液对免疫抑制小鼠免疫功能的影响

目的:研究灵芝多糖/硒化卡拉胶口服液对环磷酰胺诱导免疫抑制小鼠的非特异性免疫功能、体液免疫功能、细胞免疫功能的影响。方法:小鼠腹腔注射80mg/kg环磷酰胺诱导免疫抑制小鼠模型,灵芝多糖/硒化卡拉胶口服液连续供应30d,每天1次。测定小鼠单核巨噬细胞吞噬功能、腹腔巨噬细胞吞噬鸡红细胞功能、外周血白细胞数目、NK细胞活性、白介素-1(IL-1)活性、白介素-2(IL-2)活性、TNF-α活性、半数血清溶血素、抗体生成细胞、T、B淋巴细胞增殖能力、迟发型变态反应。结果:灵芝多糖/硒化卡拉胶口服液各剂量组(低剂量组:灵芝多糖2.5mg/kg+硒5μg/kg;中剂量组:灵芝多糖5mg/kg+硒10μg/kg;高剂量组:灵芝多糖15mg/kg+硒30μg/kg)均可提高外周血白细胞数目、提高半数溶血素值,低剂量组增强IL-1活性,中剂量组增强IL-2活性,高剂量组和中剂量组增强T、B淋巴细胞的增殖能力、NK细胞活性、TNF-α活性,高剂量组增强腹腔巨噬细胞吞噬功能、碳粒廓清值、提高抗体生成细胞、增强迟发型变态反应。结论:灵芝多糖/硒化卡拉胶口服液对免疫抑制小鼠的免疫功能具有改善作用。     

决明子蒽醌苷对小鼠免疫功能的调节作用

目的研究决明子蒽醌苷（SCAG）体外给药对小鼠细胞免疫功能的影响。方法将小鼠淋巴细胞或巨噬细胞与不同浓度的SCAG共培养，用二甲氧唑黄（XTT）法测定T及B淋巴细胞的增殖能力、对混合淋巴细胞反应（MLR）的影响、对丝裂霉素C所致淋巴细胞增殖抑制的拮抗作用，用中性红染色法观察SCAG对巨噬细胞吞噬功能、自然杀伤（NK）细胞活性、小鼠脾细胞分泌肿瘤坏死因子（TNF）活性的影响。结果SCAG体外给药可明显促进小鼠T及B淋巴细胞的增殖，增强巨噬细胞吞噬中性红的能力，提高NK细胞活性及分泌TNF活性，并可促进MLR，拮抗丝裂霉素C对淋巴细胞增殖的抑制作用。结论SCAG体外给药可显著增强小鼠细胞免疫功能。     

参苓胶囊对氟尿嘧啶增效减毒作用的研究

目的:研究参苓胶囊与氟尿嘧啶(FU)配伍对小鼠移植肿瘤S180,H22,Lewis抑瘤作用及对FU所致小鼠免疫功能抑制的保护作用.方法:按照抗癌药物筛选规程进行体内抑瘤实验;采用FU制备免疫功能低下模型,测定参苓胶囊与FU配伍对小鼠白细胞计数、腹腔巨噬细胞吞噬功能及NK细胞活性的影响.结果:不同剂量的参苓胶囊与FU配伍用均能明显提高FU对小鼠移植肿瘤S180,H22,Lewis的抑瘤率,其中高剂量组抑瘤效果最佳,配伍且能明显拮抗FU所致白细胞下降、腹腔巨噬细胞功能降低及NK细胞活性降低等毒副作用.结论:参苓胶囊与FU配伍能有明显提高FU抗肿瘤及降低其毒副作用.     

地黄多糖b的免疫抑瘤作用及其机理

从地黄中提取分离的多糖成分地黄多糖b(RPS-b),ip或ig给药抑制实体瘤S180的生长,ip给药对Lewis肺癌,B16黑素瘤和H22肝癌亦有效,最适有效剂量都在20mg·kg~(-1)体外实验证明,RPS-b对S180和HL60细胞生长均无明显的直接细胞毒作用.RPS-b在发挥抑瘤作用过程中,能提高S180荷瘤小鼠脾脏T淋巴细胞的增殖能力,并较长时间维持在较高水平;也能部分阻碍瘤株对脾脏天然杀伤细胞(NK)活力的抑制作用.说明RPS-b是一种免疫抑瘤的活性成份,其抑瘤作用是依赖于机体防御系统而间接产生的,增强机体的细胞免疫功能是其作用的重要机理     

枸杞多糖联合氟尿嘧啶对荷瘤小鼠免疫功能影响的实验研究

目的探讨枸杞多糖单独应用及与氟尿嘧啶联合应用于荷S180小鼠的抗肿瘤机制。方法小鼠右前侧腋窝接种S180肿瘤细胞,制造荷瘤小鼠模型。实验分为生理盐水组、氟尿嘧啶组、枸杞多糖高、中、低剂量组以及枸杞多糖加氟尿嘧啶组。实验结束后,检测各干预组脾指数、瘤重、肿瘤抑制率,MTT法检测淋巴细胞增殖反应、LDH检测NK细胞毒作用及中性红实验评估巨噬细胞吞噬能力,Griess法检测NO浓度,ELISA法测定TNF-α水平。结果枸杞多糖抑制荷S180小鼠肿瘤生长,刺激脾细胞增殖反应,提高NK细胞细胞毒效应以及巨噬细胞吞噬能力,刺激巨噬细胞NO分泌量,提高血清TNF-α水平。与氟尿嘧啶联合应用降低了氟尿嘧啶导致的免疫系统损伤。结论枸杞多糖可能是化疗药物的有效辅助药物。     

抗癌胶囊对实验性肿瘤的治疗及对化疗减毒作用研究

目的：研究抗癌胶囊对小鼠移植肿瘤S180，H22，Lewis的抑瘤作用及对化疗药所致小鼠免疫功能抑制的保护作用。方法：按照抗癌药物筛选规程进行体内抑瘤实验：采用氟尿嘧啶（FU）制备免疫功能低下模型，测定抗癌胶囊对小鼠白细胞计数、免疫器官重量、腹腔巨噬细胞吞噬功能和NK细胞的影响。结果：抗癌胶囊对小鼠移植肿瘤S180，H22，Lewis均有不同的抑瘤作用，其中高剂量组抑瘤效果最佳，并对小鼠体重生长无明显影响。抗癌胶囊还有明显拮抗FU所致白细胞下降，胸腺、脾脏萎缩，腹腔巨噬细胞功能降低和NK细胞减少等毒副作用。结论：抗癌胶囊具有明显的抗肿瘤作用。     

拟康氏木霉胞外多糖抗肿瘤活性研究

目的：研究拟康氏木霉胞外多糖体内外抗肿瘤活性。方法：体外采用MTT法检测拟康氏木霉胞外多糖对人白血病细胞HL一60抑制作用;体内试验是将昆明种小鼠接种S180实体瘤后随机分组,灌胃50、100、200mg·kg-1·d-1剂量的多糖,以生理盐水和环磷酰胺为空白对照组和阳性对照组。检测受试药物对小鼠S180实体瘤的抑制作用,计算小鼠的相对生长率及其免疫器官指数。结果：经MTT法检测,拟康氏木霉胞外多糖0．2～1．0mg／mL五个浓度对HL一60细胞均有不同程度的抑制作用,并呈现出剂量相关性,其中1．0mg／mL多糖浓度在72h的抑制率可达40．36％。而该多糖低、中、高三个剂量组均能明显减少S180小鼠的瘤重,其中高、中剂量组能明显提高小鼠的胸腺指数和脾指数（P〈0．05,P〈0．01）,但与空白对照组比较,三个给药组对小鼠的相对生长情况并无明显影响。结论：拟康氏木霉胞外多糖具有一定的抗肿瘤作用。     

Cancer immunotherapeutic effects of novel CpG ODN in murine tumor model

While CpG oligodeoxynucleotides (ODN) are excellent candidates for cancer immunotherapeutics, the numbers of usable CpG ODNs are limited in current clinical settings. To resolve this, we investigated whether novel CpG ODN (KSK-CpG) would be an effective immunotherapeutic in a murine tumor model by affecting in vivo and in vitro parameters, such as survival span, the number of tumor nodules, natural killer (NK) cell and cytotoxic T lymphocyte (CTL) activity and interleukin (IL)-6 or IL-12 cytokine release in splenocytes. We found that KSK-CpG was effective in the murine cancer model by way of prolonging survival span, reducing the number of tumor nodules, augmenting NK cell and CTL cytotoxicity, as well as evoking IL-6 and IL-12 cytokine release in splenocytes. Collectively, these data demonstrate that KSK-CpG is active against the highly malignant B16BL6 and EL4 tumor mouse model via innate immune augmentation.     

黄芪皂苷Ⅳ对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞功能的影响

目的：探讨黄芪皂苷Ⅳ（ASI）对小鼠T,B淋巴细胞增殖和腹腔巨噬细胞分泌的细胞因子的影响。方法：采用MTT法检测T,B淋巴细胞的增殖;采用分光光度计测定法检测抗体活性;IL－1活性用胸腺细胞增殖法测定;TNF－α活性用L9292细胞杀伤法测定。结果：1）ASI50－200mg/kg ig7天能够促进T淋巴细胞增殖和抗体生成,而ASI50－100mg/kg 能够促进B淋巴细胞增殖,但是200mg/kg对B淋巴细胞增殖无影响;（2）ASI体外仅在200nmol/L对T,B淋巴细胞有促进作用;（3）ASI 1nmol/L可以促进腹腔巨噬细胞分泌IL－1,而100－1000nmol/L则抑制腹腔巨噬细胞分泌IL－1;（4）ASI体外可以抑制LPS刺激或无LPS刺激下的腹腔巨噬细胞分泌TNF－α。结论：ASI能够促进小鼠T,B淋巴细胞的增殖和抗体生成,同时可以抑制腹腔巨噬细胞体内分泌IL－1和TNF－α。     

Immunomodulation by ethanolic extract of Boerhaavia diffusa roots

We have earlier reported that ethanolic extract of Boerhaavia diffusa, a plant used in Indian traditional system of medicine, significantly inhibits the cell proliferation. This led us to evaluate the immunomodulatory properties of this plant extract on various in vitro tests such as human natural killer (NK) cell cytotoxicity, production of nitric oxide (NO) in mouse macrophage cells, RAW 264.7, interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha), intracytoplasmic interferon-gamma (IFN-gamma) and expression of various cell surface markers on human peripheral blood mononuclear cells (PBMCs). Ethanolic extracts of B. diffusa roots inhibited human NK cell cytotoxicity in vitro, production of NO in mouse macrophage cells, IL-2 and TNF-alpha in human PBMCs. Intracytoplasmic IFN-gamma and cell surface markers such as CD16, CD25, and HLA-DR did not get affected on treatment with B. diffusa extract. Our study demonstrates immunosuppressive potential of ethanolic extract of B. diffusa.     

Synthesis and antiproliferative activity in vitro of new propargyl thioquinolines

The series of new 3,4-disubstituted thioquinolines which possess one or two O, S, Se-propargyl groups has been synthesized on the basis of the reaction of thioquinanthrene with alkoxides. All the compounds obtained were tested for their antiproliferative activity in vitro against the cells of three human cancer cell lines: SW707 (colon cancer), T47D (breast cancer), and HCV29T (bladder cancer). Two compounds, 4-(3-hydroxypropoxy)-3'-propargylthio-3,4'-diquinolinyl sulfide (3) and 3-methylthio-4-propargylselenoquinoline (13) exhibited significant cytostatic activity (ID50 < 4 micrograms/ml) against the cells of all the human cancer lines used and are good candidates for further anticancer activity studies in vitro using a broad panel of human and murine cell lines and for in vivo preclinical screening in different mouse transplantable tumor models.     

Effects of chronic dextromethorphan administration on the cellular immune responses in mice

We examined the chronic effect of dextromethorphan (DM) on the cellular immune responses in mice. T cell stimulator, phytohemagglutinin did not show significant effect on lymphocyte proliferation. Costimulator of T and B cell, pokeweed mitogen, and B cell stimulator, lipopolysaccharide exhibited DM-induced decreased lymphocyte proliferation. Significantly suppressed natural killer (NK) cell cytotoxicity was evidenced following 6 months DM exposure. These results suggest that chronic DM administration perturb B cell functioning and NK cell cytotoxicity. In addition, prenatal DM exposure did not potentiate the immunomodulation in postnatal effect induced by chronic DM.     

Eudesmin inhibits tumor necrosis factor-α production and T cell proliferation

Possible antiinflammatory effects of eudesmin were examined by assessing the effects on tumor necrosis factor (TNF)-alpha production and lymphocyte proliferation as well as cytotoxicity against murine and human macrophages. The compound significantly inhibited TNF-alpha production by lipopolysaccharide (LPS)-stimulated murine macrophage RAW264.7 without displaying cytotoxicity suggesting that eudesmin may inhibit TNF-alpha production without any interference of normal cell function. It also significantly attenuated T cell proliferation stimulated by concanavalin A (Con A) in a dose-dependent manner.     

Immunomodulatory effect of arctigenin, a lignan compound, on tumour necrosis factor-alpha and nitric oxide production, and lymphocyte proliferation

We have investigated the immunomodulatory effects of arctigenin, a dibenzyl butyrolactone lignan compound, on tumour necrosis factor (TNF)-alpha and nitric oxide (NO) production, and lymphocyte proliferation. Arctigenin inhibited strongly TNF-alpha production by lipopolysaccharide-stimulated murine macrophage RAW264.7 and differentiated human macrophage U937 with IC50 values of 5.0 and 3.9 microM, respectively, without displaying cytotoxicity. The TNF-alpha inhibitory effect of arctigenin in lipopolysaccharide-triggered RAW264.7 cells was increased by co-treatment with several known TNF-alpha inhibitors. It also potently attenuated T and B cell proliferation stimulated by concanavalin A and lipopolysaccharide in a dose-dependent manner with IC50 values of 2.9 and 14.6 microM, respectively. In contrast, the compound showed a different pattern in lipopolysaccharide- and interferon (IFN)-gamma-induced NO production from RAW264.7 cells. Arctigenin inhibited NO release by IFN-gamma signal, whereas it significantly enhanced lipopolysaccharide-triggered NO production in RAW264.7 cells. The results suggested that arctigenin may regulate immune responses in activated macrophages and lymphocytes including TNF-alpha and NO production and lymphocyte proliferation.     

Total flavonoids of Daphne genkwa root significantly inhibit the growth and metastasis of Lewis lung carcinoma in C57BL6 mice

Daphne genkwa root has been traditionally used as an effective remedy to treat various tumors. However, the active constituents for its antitumor potency have not been well documented. During the screening for antitumor constituents, it was found that the total flavonoids of D. genkwa root (TFDR) were responsible for the inhibition of tumor growth and metastasis. In this study, TFDR was investigated for its chemical composition and activities against tumor growth and metastasis. HPLC indicates that daphnodorin B, containing 42.79% of the total, represents the predominant constituent in TFDR. Treatment of LLC-bearing mice with TFDR evidently protected peripheral lymphocytes from tumor-induced reduction, and increased lymphocyte proliferation potential and cytolytic activity of NK, and inhibited tumor progression and metastasis either 7 days before, or simultaneous with, or 7 days after LLC transplantation. TFDR also suggested higher cytotoxicity to a number of tumor cell lines than that to normal human kidney cell K293. TFDR also induced an enhancement on peripheral release of TNF-alpha at doses between 25 and 75 mg/kg. These results indicated that TFDR inhibited tumor growth and metastasis by protecting host immunocyte viability and its proliferation potential, and selectively inhibiting tumor cell proliferation, and improving cytolytic activity of NK cells, and enhancing TNF release in LLC-bearing mice. Daphnodorin B and its analogues in TFDR are the active constituents in the roots of D. genkwa, contributing to the inhibition of tumor growth and metastasis.     

Effects of mycotoxins on human immune functions in vitro.

Immunosuppressive and carcinogenic Fusarium mycotoxins may appear in domestic food products. Therefore, the immunological effects of Fusarium mycotoxins were tested on human peripheral blood mononuclear cells from different blood donors. In the present study we investigated deoxynivalenol (DON), 3-acetyldeoxynivalenol, fusarenon-X, T-2 toxin, zearalenone, alpha-zearalenol, beta-zearalenol and nivalenol for their effects on T and B cells in a proliferation assay, antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) cell activity on human peripheral blood mononuclear cells. The concentrations applied in our experiments were similar to those which can be found in normal human peripheral blood system (0.2--1800 ng/ml). Among the eight mycotoxins tested, T-2 toxin, fusarenon X, nivalenol and deoxynivalenol exerted the highest immunosuppressing effect on human peripheral blood mononuclear cells in vitro. Mycotoxin-induced immunosupression was manifested as depressed T or B lymphocyte activity. Furthermore, by virtue of inhibition of NK cell activity, the protection against tumor development may also be attenuated.