大鼠外周血自然杀伤细胞对肝细胞的细胞毒效应及大黄？…

运用３Ｈ－ＴｄＲ肝细胞内掺入法对大鼠外周血自然杀伤对肝细胞的细胞毒作用进行观察，证实自然杀伤细胞对肝细胞具有细胞毒作用（Ｐ〈０．００１）。大黄在１００μｇ／ｍｌ浓度水平能抑制此自然杀伤细胞的细胞毒作用（Ｐ〈０．０５）。     

肾细胞癌bc1－2、P53、ER、PR的表达及临床意义

为了更好地选择估价肾细胞癌预后的指标，应用免疫组化技术检测５４例肾细胞癌中ｂｃ１－２、Ｐ５３蛋白及雌、孕激素受体的表达。ｂｃ１－２阳性３８例（７０．４％），Ⅰ、Ⅱ级肿瘤阳性３３例，Ⅲ、Ⅳ级肿瘤阳性５例（Ｐ＜０．０５）；Ｐ５３蛋白阳性１０例（１８．３％），对照组无一例阳性（Ｐ＜０．０５）；ＥＲ、ＰＲ阳性分别为２９例（５７．７％）、３９例（７３．７％）。ｂｃ１－２蛋白表达与ＥＲ、ＰＲ呈正相关（Ｐ＜０．０１，ｒ＝０．４５２），与Ｐ５３蛋白表达呈负相关（Ｐ＜０．０１，ｒ＝－０．４２１）。结果提示：ｂｃ１－２蛋白的表达有助于肾细胞癌预后的评估     

大块肝切除后急性肝损伤治疗的研究

将６０只大鼠随机分为２组，一组腹腔内注射肝细胞生长因子（ＨＧＦ）和１，６二磷酸果糖（ＦＤＰ），另一组代之以生理盐水做为对照，分别于术后３、７、１４天检测血清酶学变化、胃泌素含量和体外肝细胞培养蛋白质合成及ＤＮＡ合成。结果发现：治疗组大鼠术后３天ＡＬＴ较对照组迅速降低（Ｐ＜０．０１）；血浆脯肽酶（ＰＬＤ）在术后７天、１４天低于对照组（Ｐ＜０．０５）；血清胃泌素测定在术后３天、７天治疗组高于对照组（Ｐ＜０．０１）；肝细胞体外原代培养￣３Ｈ－亮氨酸掺入法显示治疗组术后３天蛋白质合成明显高于对照组（Ｐ＜０．０１）；￣３Ｈ－ＴｄＲ掺入肝细胞ＤＮＡ合成，治疗组术后各期均非常显著高于对照组（Ｐ＜０．０１）。结果证实大鼠大块肝切除后应用ＨＧＦ和ＦＤＰ，对急性肝损伤有重要的治疗作用。     

栀子对急性胰腺炎时胰腺细胞膜功能的影响

本文观察了栀子对大鼠急性胰腺炎时胰腺细胞膜功能的影响。结果显示：（１）模型组胰腺细胞膜上的Ｎａ＋－Ｋ＋－ＡＴＰ酶和Ｃａ２＋－Ｍｇ２＋－ＡＴＰ酶活性均明显低于正常组（Ｐ＜０．０１），用栀子治疗可使其保持正常。（２）治疗组的ｃＡＭＰ虽无明显变化，但ｃＧＭＰ明显降低，ｃＡＭＰ／ｃＧＭＰ比值显著增加（Ｐ＜０．００１）。（３）模型组血清乳酸脱氢酶活性明显增高，而胰腺组织中的活性则降低，治疗组与正常组相近。（４）胰腺炎时血清淀粉酶活性显著增高（Ｐ＜０．０１），治疗组与正常组很接近。上述结果表明栀子对大鼠急性胰腺炎时胰腺细胞膜的功能具有保护作用。     

肝细胞生长因子受体在人肝细胞癌中表达的研究

作者应用分子杂交法，研究肝细胞生长因子受体（ｃ－ｍｅｔ）ｍＲＮＡ在人肝细胞癌中的表达及其意义。Ｎｏｒｔｈｅｒｎ杂交分析结果表明：在癌组织中有１６例ｃ－ｍｅｔ表达阳性，在癌周肝中有１２例ｃ－ｍｅｔ表达阳性。ｃ－ｍｅｔ在肝细胞癌组织中与癌周肝组织中的表达阳性的例数差异无显著意义（Ｐ＞０．０５）。经统计学处理发现ｃ－ｍｅｔ和肝细胞癌分化程度、分期、大小、ＨＢｓＡｇ、ＡＦＰ、门静脉癌栓间无显著相关（Ｐ＞０．０５）。作者认为ｃ－ｍｅｔ基因可能在肝癌形成及转移过程中起调节作用。     

中药复方抗癌方抑制人结肠癌细胞株端粒酶活性

目的　了解中药复方抗癌方（ＫＡＦ）对人结肠癌细胞端粒酶活性的影响。方法　将ＨＣ８６９３ 细胞株作为靶细胞,用ＭＴＴ试验测定细胞毒作用,用ＴＲＡＰ半定量法测定端粒酶活性。结果　ＫＡＦ对ＨＣ８６９３ 的半数致死量（ＬＤ５０） 为２５－４％ ;染毒后,端粒酶活性降低（５％ ＫＡＦ组：４６７６－７±４７８．１ ;１０ ％ ＫＡＦ组：３５８１－３±６３２ ．４;空白对照组：６００３．３ ±３３７．９。Ｐ＜０－０５ 或Ｐ＜ ０－０１） ,去除ＫＡＦ２４ 小时后,端粒酶活性仍维持在较低水平（４４９５．０ ±４２３－９,Ｐ＜０－０５） 。结论　ＫＡＦ能抑制人结肠癌细胞端粒酶活性。     

骨纤维结构不良中癌基因蛋白的表达及意义

了解骨纤维结构不良中癌基因蛋白的表达情况。方法应用免疫组化法对３７例骨纤维结构不良行多种癌基因蛋白检测。结果３７例实验组中ｃ－ｆｏｓ、ｃ－ｍｙｃ蛋白均有高表达（Ｐ＜０．０１）。ｒａｓｐ２１蛋白在该病恶变组中有高表达（Ｐ＜０．０５）。结论ｃ－ｆｏｓ、ｃ－ｍｙｃ蛋白过度表达在骨纤维结构不良的发生发展中起重要作用，ｒａｓｐ２１蛋白过度表达在该病的恶性变过程中起作用。本研究结果对骨纤维结构不良的诊断和治疗具有广阔的应用前景。     

常用麻醉性镇痛药对家兔支气管平滑肌腺苷酸环化酶和磷酸二酯酶活力的影响

常用麻醉性镇痛药吗啡、哌替啶和芬太尼对家兔离体支气管平滑肌腺苷酸环化酶（ＡＣ）和磷酸二酯酶（ＰＤＥ）活力的影响，结果表明：吗啡可使ＡＣ的活性由对照组的１５．９９１１±０．８５７９ｕ降至１５．０４６３±０．７９８９ｕ，Ｐ＜０．０５．在吗啡加纳络酮组，这种对ＡＣ的抑制作用依然存在。芬太尼使ＰＤＥ的活性由对照组的３．４８９２±０．７１２３ｕ上升至４．３０５４±０．７６７４ｕ，Ｐ＜０．０５。实验结果提示：（１）吗啡可减少支气管平滑肌中的ｃＡＭＰ生成，芬太尼可增加ｃＡＭＰ的降解，致使ｃＡＭＰ含量减少，这可能与其增加支气管平滑肌张力的作用有关；（２）吗啡对支气管平滑肌ＡＣ的抑制作用不受纳络酮的影响，提示该作用可能不是通过阿片受体起作用的，作用焦点可能在Ｇ－蛋白或催化亚单位。     

凋亡相关基因产物Fas/APO-1和bcl-2蛋白在肾癌组织中的表达及意义

目的研究凋亡相关基因Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２在肾癌发生发展中的作用。方法采用免疫组织化学法对３５例肾癌组织和２６例远离肾癌的正常肾组织Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２蛋白的表达进行检测。结果肾癌组织Ｆａｓ／ＡＰＯ－１蛋白表达率为５７．１４％，明显低于正常肾组织中的表达率（８４．６２％，Ｐ＜０．０５），且表达强度也明显低下；而ｂｃｌ－２蛋白表达率为８０．００％，明显高于正常肾组织中的表达率（５３．８５％，Ｐ＜０．０５）。结论Ｆａｓ／ＡＰＯ－１与ｂｃｌ－２基因共同参与了肾癌的发生和发展。     

化学发光法检测血液透析患者细胞免疫功能的研究

通过化学发光技术对血液透析患者的吞噬细胞的吞噬功能、致敏淋巴细胞和自然杀伤细胞活化程度进行了研究，结果表明，血液透析患者的吞噬细胞吞噬功能、吞噬细胞的预激活状态均高于正常人，具有非常显著性差异（Ｐ＜０．００１），而血浆调理活性则无显著性差异（Ｐ＞０．０２），淋巴细胞活化程度高于正常人，具有非常显著性差异（Ｐ＜０．００１），自然杀伤细胞活性程度高于正常人，具有显著性差异（Ｐ＜０．０２）。故化学发光法对检测血液透析患者的机体免疫功能，了解各种细胞的免疫状态有一定的实际意义。     

Effects of interferons on syngeneic murine malignant glioma-specific killer T cell

The effects of IFN (interferon) on the activation and differentiation of syngeneic murine malignant glioma-specific killer T cell were investigated in C57BL/6 adult mice in order to clarify the potential usefulness for anti-tumor local immunotherapy. It was confirmed that the specific killer T cell against 20-methylcholanthrene-induced 203-glioma was generated in mice after intracranial and subcutaneous inoculation of the tumor cells. The cytotoxic activities of splenic cells obtained from intracranial and subcutaneous tumor-bearing mice were assessed on MLTC (mixed lymphocyte-tumor cell culture) for 18 hours by microcytotoxicity assay modified Takasugi and Klein's method. The addition of IFN to MLTC resulted in a similar 1.5- to 2.0-fold increase of generated cytotoxic activities. Prior treatment of sensitized splenic cells with IFN resulted in an enhancement of the specific cytotoxic activity for the target tumor cells. IFN enhanced cytotoxic activities in MLTC only in the first 3 hours. These cytotoxic activities were eliminated by the treatment of sensitized lymphocytes with anti-Thy-1 antibody and complement before adding IFN. Therefore, it was found that IFN was able to enhance killer T cell activity of sensitized lymphocytes. Normal lymphocytes did not exhibit any cytotoxic activity even after the treatment with IFN. On the other hand, IFN had a cytostatic effect on the growth of 203-glioma cells. This effect of IFN on 203-glioma cells was not observed when IFN was removed from the suspension (by washing 203-glioma cells).(ABSTRACT TRUNCATED AT 250 WORDS)     

Augmentation of cytotoxic activity by combination with interleukin 2 and interferon gamma

The synergy of cytotoxic activity by interleukin 2 (IL-2) and interferon gamma (IFN-gamma) was evaluated in human peripheral blood mononuclear cells (PBMC) and spleen cells. PBMC incubated with IL-2 (10 IU/ml) and IFN-gamma (200 IU/ml) for 4 days showed the stronger cytotoxic activity against K562, MOLT-4 and Daudi cells. Combination with IL-2 and IFN-gamma induced stronger activity than IL-2 or IFN-gamma alone. In order to investigate the sequential roles of IL-2 and IFN-gamma in the killer cell function, the cells were stimulated with IFN-gamma after washing of IL-2 or stimulated by IFN-gamma at various timing and duration without washing of IL-2. IL-2 was essential to induce the synergistic effect of IL-2 and IFN-gamma to cytotoxic activity. The similar augmentation of cytotoxic activity was observed by the addition of IFN-gamma at any incubation periods with IL-2, compared with stimulation with IL-2 or IFN-gamma alone. The phenotypes of the killer cells by stimulation with IL-2, IFN-gamma alone or IL-2 plus IFN-gamma were mainly CD2+, CD16+, indicating the activated natural killer cells.     

Mechanisms of action of bacillus Calmette-Guérin in the treatment of superficial bladder cancer

Summary Local immunotherapy with bacillus Calmette-Guérin (BCG) is an effective treatment to prevent recurrence and progression of superficial bladder cancer, but the antitumor mechanism of action of BCG remains unclear. There are some experimental and clinical data suggesting that BCG antigens are processed not only by immunocompetent cells but also by urothelial cells and tumor cells. The foreign antigen may be presented at the cell curface by major histocompatibility complex (MHC) class II molecules and recognized by CD4 cells. The cytotoxic effect could result from the direct activity of CD4 cells or from the cytotoxic effect of released cytokines and the activation of other cytotoxic cells [cytolytic T-lymphocytes CTLs; CD8 cells), macrophages, natural killer (NK) or lymphokine-activated killer (LAK) cells]. These mechanisms are also involved in tumorrejection, and the identification of some specific tumor rejection antigens presented to specific CTLs could provide new therapeutic approaches.     

ICOS基因转染对CIK细胞杀伤胆管癌细胞作用的研究

目的 探讨转染可诱导共刺激分子(inducible co-stimulator, ICOS)基因对细胞因子诱导杀伤(cytokine. induced killer, CIK)细胞杀伤胆管癌细胞的作用.方法 构建含ICOS基因的腺病毒载体并转染给CIK细胞(CIK-ICOS细胞组),单纯CIK及CIK-EGFP细胞为对照组,观察3组CIK细胞体外增殖与凋亡情况以及对胆管癌细胞的杀伤作用;ELISA法检测3组CIK细胞上清液中IFN-γ、IL-2及TNF-α的表达.建立胆管癌移植瘤的SCID小鼠模型,按随机数字表法将40只SCID小鼠分为生理盐水对照组、CIK、CIK-EGFP、CIK-ICOS细胞治疗组,观察转染ICOS基因的CIK细胞对小鼠胆管癌细胞的杀伤作用.结果 CIK-ICOS细胞组体外增殖明显强于CIK及CIK-EGFP细胞组;培养第20天CIK-ICOS与CIK细胞凋亡率分别为0.69%和2.90%,第23天分别为0.89%和4.92%;在不同效靶比CIK-ICOS细胞组杀伤效应均显著高于CIK与CIK-EGFP细胞组(F=13.37,6.46,25.51,P<0.05);CIK-ICOS细胞组上清液中IFN-γ的浓度为(49.50±4.73)μg/L,显著高于CIK细胞组(30.53±3.73)μg/L及CIK-EGFP细胞组(30.12±2.64)μg/L(F=38.89,P<0.05).CIK-ICOS细胞治疗组小鼠肿瘤生长速度明显低于CIK与CIK-EGFP细胞治疗组,肿瘤坏死面积明显大于CIK与CIK-EGFP细胞治疗组,瘤内CIK细胞数量最多.结论 CIK细胞在体内外均具有杀伤胆管癌细胞的作用.转染ICOS基因后,CIK细胞体外存活时间延长、增殖能力增强、IFN-γ的表达增多,其在体内外抗胆管癌作用明显增强.     

Antitumor activity against established intracerebral gliomas exhibited by cytotoxic T lymphocytes, but not by lymphokine-activated killer cells.

Specific immune responses against malignant brain tumors have been difficult to demonstrate. Moreover, immunotherapy has met with little success, despite using lymphocytes with high levels of cytotoxicity against brain tumor cells. Lymphokine-activated killer (LAK) cells that nonspecifically kill brain tumor cells are produced by stimulating resting precursors with high concentrations of interleukin-2 (IL-2). Cytotoxic T lymphocytes that specifically kill brain tumor cells are produced by stimulating antigen receptor-positive immune-cell precursors with tumor cells. In an attempt to gain insight into immune cell function against brain tumors, the present study compared the in vitro and in vivo activities of LAK cells and cytotoxic T lymphocytes produced against RT2, a fast-growing rat glioma cell line. Lymphokine-activated killer cells were produced by stimulating normal rat spleen cells with 1000 units of IL-2, and RT2-specific cytotoxic T lymphocytes were produced by priming them in vivo with RT2 and Corynebacterium parvum and restimulating primed spleen cells with RT2 in vitro. Lymphokine-activated killer cells were highly cytotoxic for a panel of syngeneic and allogeneic brain tumor and non-brain tumor target cells, including RT2, as measured in a 4-hour 51Cr release assay. Cytotoxic T lymphocytes were highly cytotoxic only for syngeneic brain tumor target cells. Lymphokine-activated killer cells and cytotoxic T lymphocytes were tested for in vivo antitumor activity against intracerebral RT2 by intravenous adoptive transfer of activated lymphocytes. Untreated rats died in approximately 2 weeks. Lymphokine-activated killer cells plus IL-2 failed to affect survival when treatment was initiated as early as 1 day following tumor inoculation. Cytotoxic T lymphocytes and IL-2 administered as late as Day 5 rejected progressing intracerebral tumor. Thus, although both cytotoxic T lymphocytes and LAK cells exhibited high levels of in vitro killing of glioma cells, only cytotoxic T lymphocytes rejected progressing intracerebral tumors.     

乳腺癌树突状细胞瘤苗对自体CIK细胞生物活性的影响

目的探讨乳腺癌树突状细胞瘤苗（DCs）对自体CIK细胞生物活性的影响。方法取诱导7d的活化DCs瘤苗和诱导7d的自体CIK细胞混合培养（DC—CIK细胞），同期自体CIK细胞为对照组。检测2、8、14d的DC—CIK细胞CD3与CD56表型、培养上清液IL．12的水平及测定混合培养8d时对乳腺癌细胞MCF．7的细胞毒效应。结果混合培养14d的DC-CIK细胞增殖率显著高于CIK细胞（P〈0．01），但CD3和CD56表型无显著变化（P〉0．05）。DC—CIK细胞对MCF-7乳腺癌细胞的特异性溶解率和细胞毒活性均高于CIK细胞（P〈0．01）：CIK细胞强烈刺激DCs分泌IL-12．混合培养2d时，DCs的IL-12p40分泌水平达到高峰。结论乳腺癌DCs瘤苗与自体CIK细胞混合培养后（DC—CIK细胞），进一步促进了DCs的成熟并增强了CIK细胞的细胞毒活性。     

微囊豚鼠肝细胞培养及其细胞免疫屏障作用的观察

目的 观察海藻酸钠-多聚赖氨酸-海藻酸钠(APA)微囊化肝细胞的生物学功能及对免疫细胞的隔离效应。方法 用胶原酶门腔静脉灌注法分离豚鼠肝细胞,测定微囊化肝细胞及游离肝细胞培养上清白蛋白水平,用自然杀伤细胞(NK)细胞的细胞毒实验来评价微囊的免疫隔离效应。结果 微囊包裹对肝细胞的活率无明显影响,微囊化肝细胞与裸露肝细胞白蛋白分泌水平差异无显著性(P>0.05),NK细胞对K562靶细胞的细胞毒实验表明微囊可有效地保护囊内细胞不受NK细胞的杀伤作用。结论 APA微囊化肝细胞可保持良好的生物活性,APA微囊对细胞免疫具有免疫隔离作用。     

Altered cellular immune function in the atelectatic lung

Pulmonary atelectasis is common and may predispose the lung to infection. We have previously shown that atelectasis impairs alveolar macrophage antibacterial function. This study examines the effect of atelectasis on the cytotoxic function of lymphocytes harvested from the bronchoalveolar space of atelectatic lung segments by bronchoalveolar lavage. Specifically, we studied natural killer and lectin-dependent cell-mediated cytotoxicity in peripheral blood and bronchoalveolar lavage lymphocytes from the atelectatic lower lobes and contralateral normal lobes in a group of 8 dogs. We observed a decline of natural killer and lectin-dependent cell-mediated cytotoxicity to 62.7% and 61.5%, respectively, of preatelectasis control values in the affected lung lobes (p less than 0.01). Simultaneous measurements of cytotoxic activity of bronchoalveolar lavage lymphocytes harvested from the unaffected contralateral normal lungs were comparable with control values. On the other hand, natural killer and lectin-dependent cell-mediated cytotoxicity activities in peripheral blood lymphocytes were significantly increased in animals having right lower lobe atelectasis (166.7% and 154.7% of pretreated normal control, respectively, p less than 0.01). Atelectasis was also associated with an influx of polymorphonuclear leukocytes into the bronchoalveolar compartment. These findings confirm the presence of natural killer cells and cytotoxic lymphocytes in the bronchoalveolar compartment and demonstrate an atelectasis-induced impairment of local bronchoalveolar lymphocyte function. Such a dysfunction of local lung cellular host defenses may render the atelectatic lung susceptible to infection.     

Reconstitution of cytotoxic T lymphocyte activity following allogeneic bone marrow transplantation in man

Longitudinal studies over an eight-month period have been performed to follow the T killer response restoration after allogeneic bone marrow transplantation (BMT). The capacity of the patient's peripheral blood mononuclear cells (PBM) to develop cytotoxic effector cells directed either against allogeneic cells or against Epstein-Barr virus (EBV) or Herpes-simplex virus-1 (HSV-1)-infected syngeneic cells was tested monthly. The data suggest that in most cases the cytotoxic T lymphocyte (CTL) activity, either allospecific or directed against EBV and the HSV-1 specific killer (HNK) responses, is quickly reconstituted in BMT patients. Roughly, these activities seem to be normal after two months following classic nondepleted grafting. However, in the two patients who had received a T-cell-depleted graft, the restoration was delayed. Furthermore, in the majority of the patients, once reconstituted the killer responses were relatively stable and the occurrence or absence of graft-versus-host diseases (GVHD) did not significantly modify these responses.     

Immune mechanisms in acetaminophen-induced acute liver failure

An overdose of acetaminophen (N-acetyl-p-aminophenol, APAP), also termed paracetamol, can cause severe liver damage, ultimately leading to acute liver failure (ALF) with the need of liver transplantation. APAP is rapidly taken up from the intestine and metabolized in hepatocytes. A small fraction of the metabolized APAP forms cytotoxic mitochondrial protein adducts, leading to hepatocyte necrosis. The course of disease is not only critically influenced by dose of APAP and the initial hepatocyte damage, but also by the inflammatory response following acetaminophen-induced liver injury (AILI). As revealed by mouse models of AILI and corresponding translational studies in ALF patients, necrotic hepatocytes release danger-associated-molecular patterns (DAMPs), which are recognized by resident hepatic macrophages, Kupffer cell (KC), and neutrophils, leading to the activation of these cells. Activated hepatic macrophages release various proinflammatory cytokines, such as TNF-α or IL-1β, as well as chemokines (e.g., CCL2) thereby further enhancing inflammation and increasing the influx of immune cells, like bone-marrow derived monocytes and neutrophils. Monocytes are mainly recruited via their receptor CCR2 and aggravate inflammation. Infiltrating monocytes, however, can mature into monocyte-derived macrophages (MoMF), which are, in cooperation with neutrophils, also involved in the resolution of inflammation. Besides macrophages and neutrophils, distinct lymphocyte populations, especially γδ T cells, are also linked to the inflammatory response following an APAP overdose. Natural killer (NK), natural killer T (NKT) and T cells possibly further perpetuate inflammation in AILI. Understanding the complex interplay of immune cell subsets in experimental models and defining their functional involvement in disease progression is essential to identify novel therapeutic targets for human disease.