目的 探讨辛伐他汀纳米粒对脓毒症致急性肺损伤(ALI)小鼠体内Toll样受体9(TLR9)表达水平及预后的影响.方法 将所选取的小鼠分为对照组(CO组)、脓毒症组(SE组)、静脉注射组(LI组)、纳米粒组(NA组),每组13只.采用免疫印迹法检测肺组织中TLR9蛋白表达水平,记录各组小鼠7 d内生存率,计算各组小鼠湿干重比值(W/D),采用HE染色法检测各组小鼠肺组织病理形态,采用实时荧光定量PCR检测肺组织中TLR9 mRNA表达水平.结果 LI组、NA组小鼠W/D低于SE组(均P<0.05),NA组小鼠W/D低于LI组(P<0.05).与SE组、LI组比较,NA组小鼠肺体积有所减小,微血管扩张好转.SE组肺组织TLR9 mRNA表达水平高于LI组、NA组(均P<0.05),NA组小鼠肺组织中TLR9 mRNA表达水平低于LI组(P<0.05).LI组、NA组小鼠肺组织TLR9蛋白表达水平低于SE组(均P<0.05),NA组小鼠肺组织中TLR9蛋白表达水平低于LI组(P<0.05).LI组、NA组小鼠生存率高于SE组(均P<0.05),LI组小鼠生存率低于NA组(P<0.05).结论 辛伐他汀对脓毒症致ALI小鼠具有一定治疗作用,可改善小鼠肺组织病理结构,提高小鼠的生存率,其机制可能与降低TLR9信号通路表达水平有关.

目的 对老年危重症患者心衰发病临床特征及相关危险因素进行探讨.方法 将我院于2018年4月至2019年4月收治的200例老年重症患者以回顾性分析方法进行临床资料的分析.对200例患者的年龄、冠心病、高血脂、肺结核、心衰、肾功能不全等既往病史和酗酒、大型手术等的危险因素进行细致分析.结果 在本次研究中,200例患者中合并有心衰的共93例,平均年龄为(77.42±12.14)岁高于无心衰患者的(73.18±7.94)岁,P<0.05;在合并有心衰患者中23例的左室射血分小于40％,且其平均年龄(72.21±5.74)岁小于其余合并有心衰的患者平均年龄(77.12±10.94)岁,P<0.05.而根据Logistic回归分析,冠心病、糖尿病、肾功能不全、大型手术是老年重症患者心力衰竭的主要独立危险因素,P<0.05.结论 老年危重症患者出现心力衰竭情况的诱发因素有多种,而对患者进行有效的肾脏功能维护和降低大型手术频率能够减少心力衰竭的发病情况.

目的 对重症急性胰腺炎合并急性呼吸衰竭患者,采用ICU治疗方式进行治疗,观察分析临床治疗效果.方法 选取我院自2017年8月至2018年7月收治的重症急性胰腺炎合并急性呼吸衰竭患者共40例,分为常规组及ICU组,各20例.常规组的患者采用常规治疗方式,ICU组的患者采用ICU治疗方式.观察统计两组患者的治疗情况,并进行对比分析.结果 常规组的总有效率为75％,实验组95％,差异明显;常规组的平均机械通气时间、平均住院时间分别为(28.16±1.05)d、(32.11±0.16)d,ICU组分别为(15.81±1.39)d、(16.19±1.18)d,组间数据对比差距非常明显,符合统计学意义的明显差异要求.结论 使用ICU治疗方式对患者进行精心治疗,能够减轻患者的生理痛楚,缓解急性胰腺炎的症状,并能够在短时间内改善患者的呼吸功能,提升患者的生活质量,具有临床推广价值.

目的 分析铜绿假单胞菌-甘露糖敏感血凝素(PA-MSHA)预处理对严重脓毒症大鼠肠组织和腹腔灌洗液中细胞因子和血管细胞黏附分子-1(VCAM-1)、细胞间黏附分子-1(ICAM-1)表达的影响.方法 取雄性SD大鼠30只,随机分为对照组(未建立脓毒症模型)、模型组(建立严重脓毒症模型)和实验组(建立严重脓毒症模型+给予PA-MSHA预处理),三组各10只,分别进行相应处理.通过苏木精-伊红染色观察三组大鼠病理学改变,采用酶联免疫吸附试验法对三组大鼠肠道内VCAM-1和ICAM-1水平进行检测,采用相同方法检测大鼠腹腔灌洗液和肠组织中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)及白介素-10(IL-10)的水平,并进行比较.结果 对照组肠上皮细胞未见异常,肠腺体未见异常,未出现炎症细胞浸润;模型组和实验组均出现不同程度的肠上皮细胞破坏、毛细血管扩张、固有层水肿及炎症细胞浸润,但实验组肠道损伤程度较模型组明显减轻.模型组和实验组肠组织中VCAM-1和ICAM-1水平较对照组显著升高(P<0.05).模型组和实验组腹腔灌洗液中TNF-α、IL-6及IL-10的水平较对照组显著升高(P<0.05);实验组腹腔灌洗液中TNF-α、IL-6及IL-10的水平较模型组显著下降(P<0.05).模型组和实验组肠组织中TNF-α、IL-6及IL-10的水平较对照组显著升高(P<0.05);实验组肠组织中TNF-α和IL-6较模型组显著下降,IL-10的水平较模型组显著升高(P<0.05).结论 PA-MSHA预处理可促使局部细胞因子和VCAM-1及ICAM-1水平下降,从而可有效减轻脓毒症大鼠肠道炎症损害,缓解肠道炎症反应,减轻脓毒症病情进展,因此对脓毒症导致的肠道损伤具有良好的预防效果.

Purpose: Malignant melanoma's (MM) incidence is rising faster than that of any other cancer in the US and the overall survival at 5 years is less than 10%. B cell associated protein 31 (BAP31) is overexpressed in most MMs and might be a promising target for immunotherapy of this disease. Experimental design: Firstly, we investigated the expression profiles of human BAP31 (hBAP31) and mouse BAP31 (mBAP31) in human and mouse normal tissues, respectively. The expression level of hBAP31 in human MMs and mBAP31 in B16 melanoma cells was also analyzed. Then we constructed novel mBAP31 DNA vaccines and tested there ability to stimulate mBAP31-specific immune responses and antitumor immunity in B16 melanoma-bearing mice. Results: For the first time, we found that protein expression of hBAP31 were dramatically upregulated in human MMs when compared with human normal tissues. Predominant protein expression of mBAP31 was found in mouse B16 melanoma cells but not in mouse important organs. When mice were immunized with mBAP31 DNA vaccines, strong cellular response to mBAP31 was observed in the vaccinated mice. CTLs isolated from immunized mice could effectively kill mBAP31-positive target mouse B16 melanoma tumor cells in vitro and vaccination with mBAP31 DNA vaccines had potent anti-tumor activity in therapeutic model using B16 melanoma cells. Conclusions: These are the first data supporting a vaccine targeting BAP31 that is capable of inducing effective immunity against BAP31-expressing MMs and will be applicable to human MMs and hBAP31 DNA vaccine warrants investigation in human clinical trials.

目的从分泌抗SEB的单克隆抗体(FMU-SEB-No.1)杂交瘤细胞中,克隆出FMU-SEB-No.1重链和轻链可变区(VH和VL)基因,构建FMU-SEB-No.1单链抗体(scFv)的原核表达载体,并进行scFv基因的蛋白表达。方法从FMU-SEB-No.1杂交瘤细胞中提取总RNA,采用RT-PCR扩增出VH和VL基因;通过在引物上设计linker序列,拼接VH和VL为完整FMU-SEB-No.1的scFv基因(FMU-SEB-scFv)。将测序正确的scFv基因克隆入PGEX4T-1载体,转化入E.coli BL21(DE3)进行蛋白表达。通过SDS-PAGE,Western blot法分析其表达水平和特异性,酶联免疫吸附试验(ELISA)鉴定其抗原结合活性。结果测序结果显示,本实验成功克隆出FMU-SEB-No.1重链及轻链可变区基因,并成功构建FMU-SEB-scFv基因,所得的基因全长为750 bp,编码250个氨基酸。SDS-PAGE和Western blot分析表明,PGEX4T1-FMU-SEB-scFv在E.coli BL21(DE3)可表达为Mr约54 000的可溶型scFv/GST融合蛋白。间接ELISA检测结果表明,可溶型scFv/GST融合蛋白与SEB具有较高的抗原结合活性。结论制备并鉴定了针对SEB的基因工程抗体,为开发出针对SEB的治疗性抗体奠定了实验基础。

Background Cartilage degradation is a typical characteristic of arthritis. This study examined whether there was a subset of phagocytic chondrocytes that expressed the specific macrophage marker, CD163, and investigated their role in cartilage degradation. Methods Cartilage from the knee and temporomandibular joints of Sprague-Dawley rats was harvested. Cartilage degradation was experimentally-induced in rat temporomandibular joints, using published biomechanical dental methods. The expression levels of CD163 and inflammatory factors within cartilage, and the ability of CD163+ chondrocytes to conduct phagocytosis were investigated. Cartilage from the knees of patients with osteoarthritis and normal cartilage from knee amputations was also investigated. Results In the experimentally-induced degrading cartilage from temporomandibular joints, phagocytes were capable of engulfing neighboring apoptotic and necrotic cells, and the levels of CD163, TNF-α and MMPs were all increased (P<0.05). However, the levels of ACP-1, NO and ROS, which relate to cellular digestion capability were unchanged (P>0.05). CD163+ chondrocytes were found in the cartilage mid-zone of temporomandibular joints and knee from healthy, three-week old rats. Furthermore, an increased number of CD163+ chondrocytes with enhanced phagocytic activity were present in Col-II+ chondrocytes isolated from the degraded cartilage of temporomandibular joints in the eight-week experimental group compared with their age-matched controls. Increased number with enhanced phagocytic activity of CD163+ chondrocytes were also found in isolated Col-II+ chondrocytes stimulated with TNF-α (P<0.05). Mid-zone distribution of CD163+ cells accompanied with increased expression of CD163 and TNF-α were further confirmed in the isolated Col-II+ chondrocytes from the knee cartilage of human patients with osteoarthritis, in contrast to the controls (both P<0.05). Conclusions An increased number of CD163+ chondrocytes with enhanced phagocytic activity were discovered within degraded joint cartilage, indicating a role in eliminating degraded tissues. Targeting these cells provides a new strategy for the treatment of arthritis.

目的构建针对B型葡萄球菌肠毒素(SEB)的特异性单链抗体(scFv),并进行纯化和鉴定。方法设计抗SEB高亲和力单克隆抗体(mAb)FMU-SEB-No.2重链和轻链可变区基因片段VH和VL的引物,利用RT-PCR技术,从本室制备的抗SEBmAb FMU-SEB-No.2杂交瘤细胞株中扩增出VH和VL基因片段;然后通过在轻链引物上设计linker序列,重叠引物延伸法(SOE)将VH和VL基因拼接成scFv基因片段,并将其克隆入原核表达载体PGEX4T-1中;转化E.coli BL21(DE3)感受态细胞,IPTG诱导表达,融合蛋白经谷胱甘肽亲和层吸柱纯化。采用SDS-PAGE,Western blot,ELISA等方法对其表达水平和特异性进行分析。结果成功从1株抗SEB mAb FMU-SEB-No.2杂交瘤细胞株中扩增出VH和VL可变区基因,并通过linker序列将其拼接成scFv片段,大小约750 bp,编码250个氨基酸。PCR、酶切鉴定和测序结果表明表达载体构建成功。转化E.coli BL21(DE3),经IPTG诱导后,以可溶形式表达相对分子质量(Mr)约54 000的融合蛋白。经谷胱甘肽亲和层析法纯化融合蛋白,可获得纯度达90%以上的scFv,ELISA和Western blot法检测结果表明,可溶性scFv与抗原SEB有较强的结合活性。结论成功构建并表达出抗SEB的特异性单链抗体。

Deuterium quadrupolar echo NMR was applied to precision CD3 branched polyethylene at temperatures ranging from below the glass transition up to the melting point. The CD3 branches were placed on every 15th or 21st carbon with zero variation in the branch spacing by acyclic diene metathesis polymerization chemistry. The deuterium lineshapes were simulated and fit to the experimental spectrum assuming appropriate models that approximate the motions in the amorphous and crystalline phases. Spectral contributions of each phase were isolated by T1 fitting. The fitting results comprise the isotropic reorientation correlation time distribution and axial jump angle distribution in these two phases, respectively. The mean jump angle was found to increase monotonically with temperature, approaching 35° near the melting point, consistent with previous carbon-13 NMR results on this same polymer.

AIM:To construct an expression plasmid for NY-ESO-1 gene and identify the expression of recombinant protein NY-ESO-1/GST in E.coli.METHODS:NY-ESO-1 segment was amplified from the testis cDNA library by RT-PCR and cloned into the prokaryotic expression vector pGEX4T-1 downstream tagged by GST to construct the expression plasmid pGEX-4T1-NY-ESO-1. The recombinant vector was transformed to BL21 (DE3) and NY-ESO-1/GST fusion protein was induced expression by IPTG. The protein was purified by urea elution and identified by SDS-PAGE and Western blotting.RESULTS:The NY-ESO-1 segment was successfully amplified and its sequence was identical with that published in GenBank. The BL21 (DE3) pLysS containing the pGEX-4T1-NY-ESO-1 expressed a M(r); 44 000 fusion protein under the induction of IPTG. The purity of the protein was 90%. Western blotting proved that NY-ESO-1/GST had a specific reaction with anti-GST mAb.CONCLUSION:The prokaryotic expression vector of NY-ESO-1 has been constructed and the fusion protein NY-ESO-1/GST of high purity is successfully expressed.

Botulinum neurotoxins (BoNTs) are the most poisonous substances ever known. The early detection of these toxins could bear more time for appropriate medical intervention. The standard method for detecting BoNTs is the mouse bioassay, which is time consuming (up to 4 days) and requires a large number of laboratory animals. The immunologic detection methods could detect the toxins within a day, but most of these methods are less sensitive compared with the mouse bioassay due to the lack of high-affinity antibodies. Recently, the recombinant H(C) subunit of botulinum neurotoxin type A (rAH(C)) was expressed as an effective vaccine against botulism, indicating that the rAH(C) could be an effective immunogen that raises the monoclonal antibody (mAb) for detecting BoNT/A. After immunized BALB/c mice with rAH(C), 56 mAbs were generated. Two of these mAbs were selected to establish a highly sensitive sandwich chemiluminescence enzyme immunoassay (CLEIA), in which FMMU-BTA-49 and FMMU-BTA-22 were used as capture antibody and detection antibody, respectively. The calculated limit of detection (LOD) based on molecular weight of rAH(C) and BoNT/A reached 0.45 pg mL(-1). This CLEIA can be used in the detection of BoNT/A in matrices such as milk and beef extract. This method has 20-40 fold lower LOD than that of the mouse bioassay and takes only 3 h to complete the detection, indicating that it can be used as a valuable method to detect and quantify BoNT/A.

Botulinum neurotoxins (BoNTs) are classified as category A biological threat agents by the Centers for Disease Control and Prevention (CDC) in the United States for its hazardous and potential bioterrorist threat to the public. About 1% naturally occurring botulisms are caused by Botulinum neurotoxin serotype F (BoNT/F). Most of the immunoassays for detecting BoNTs focus on the serotypes A and B, but few methods have been established for the detection of BoNT/F. Recently, the recombinant Hc subunit of botulinum neurotoxin type F (rFHc) was expressed as an effective vaccine against BoNT/F, indicating that this rFHc could be an effective immunogen to raise monoclonal antibodies (MAbs) for the detection and neutralization of BoNT/F. Here we present a novel sandwich enzyme-linked immunosorbent assay (ELISA) based on two MAbs against rFHc, which were FMMU-BTF-8 and FMMU-BTF-29 as capture antibody and detection antibody, respectively. The limit of detection (LOD) of this ELISA reached 12.09 pg/mL, much less than that of the other reported immunoassays. A simple, sensitive ELISA for detecting and quantifying BoNT/F was established, which can be used as a valuable method to detect and quantify BoNT/F.

Background A number of strategies have been used to improve the efficacy of the DNA vaccine for the treatment of tumors. These strategies, ranging from activating CD4+ T cell, manipulating antigen presentation and/or processing to anti-angiogenesis, focus on one certain aspect in the functioning of the vaccine. Therefore, their combination is necessary for rational DNA vaccines design by synergizing different regimens and overcoming the limitations of each strategy. Methods A DNA fragment (HSV) encoding the C terminal 37 amino acids of human chorionic gonadotropin beta chain (hCG beta), 5 different HLA-restricted cytotoxic T lymphocyte epitopes from human survivin and the third and fourth extracellular domains of vascular endothelial growth factor receptor 2 (VEGFR2) was inserted into the sequence between the luminal and transmembrane domain of human lysosome-associated membrane protein-1 cDNA for the construction of a novel DNA vaccine. Results This novel vaccine, named p-L/HSV, has a potent antitumor effect on the LL/2 lung carcinoma model in syngeneic C57BL/6 mice. The immunologic mechanism involved in the antitumor effect referred to the activation of both cellular and humoral immune response. In addition, the tumor vasculature was abrogated as observed by immunohistochemistry in p-L/HSV immunized mice. Furthermore, the immunized mice received an additional boost with p-L/HSV 6 months later and showed a strong immune recall response. Conclusions The present study indicates that the strategies of combining antitumor with antiangiogenesis and targeting the tumor antigen to the major histocompatibility complex class II pathway cooperate well. Such a study may shed new light on designing vaccine for cancer in the future. Copyright (c) 2012 John Wiley & Sons, Ltd.

Natural killer (NK) cells play a critical role in antitumor immunity, and the activation of NK cells is regulated by a series of NK cell receptors. Here, we show that crosslinking CD226, an important NK cell receptor, with the anti‐CD226 mAb LeoA1 on NKL cells, regulated the expression of several microRNA and transmembrane tumor necrosis factor‐α. Among them, miR‐30c‐1* was noticed because overexpression of miR‐30c‐1* triggered upregulation of transmembrane tumor necrosis factor‐α expression and enhanced NK cell cytotoxicity against hepatoma cell lines SMMC‐7721 and HepG2. Furthermore, we proved that the inhibitory transcription factor HMBOX1, which depressed the activation of NK cells, was the direct target gene of miR‐30c‐1*. In conclusion, our results revealed a novel regulatory mechanism: miR‐30c‐1* promoted NK cell cytotoxicity against hepatoma cells by targeting HMBOX1. (Cancer Sci 2012; 103: 645–652)

A structural investigation of linear ethylene-co-vinyl amine (EVAm) copolymers having a primary amine branch on every 9th, 15th, 19th, or 21st carbon along the ethylene backbone has been completed using step polymerization chemistry. Acyclic diene metathesis (ADMET) polymerization has been used with symmetrical alpha,omega dienes containing protected amine groups to afford polymers with exact primary structures and constant methylene run lengths between branches. The effects of subtle structural changes such as the ethylene run lengths between amine branches can be observed and used to correlate structure property relationships. NMR and FT-IR techniques are used to characterize and verify the excellent structural control this synthetic approach provides over traditional chain polymerization techniques. Thermal decomposition of these copolymers is shown to additionally support polymer structure while differential scanning calorimetry demonstrates crystallinity in the polymers with an amine on every 15th and 21st carbon, whereas the polymer with an amine on every ninth carbon is amorphous. Variations of the physical and spectral properties are discussed as a consequence of the amine branch spacing, protection, and saturation of the ethylene backbone.

In this study,we aimed to construct a expression plasmid for△BAP31 gene and identify the recombinant protein expression.Firstly,the△BAP31 coding sequence was amplified by polymerase chain reaction(PCR) method and subcloned into pGEX-4T1vector.After the target gene was sequenced,the plasmid was transformed into E coil DH5α and induced with IPTG to express fusion protein △BAP31/GST which was then proved by SDS-PAGE and Western blot.SDS-PAGE analysis showed that a novel protein with the expected molecular mass about 40 000 was expressed with the inducement of IPTG.The fusion protein was existed mostly in the form of soluble protein,and grayscale scanning showed that the expressed △BAP31/GST fusion protein was accounted for 70.6% of the total bacterium protein.The purity of the fusion protein reached 94.5% after purification by High-Affinity GST Resin.Then western blot confirmed the recombinant protein was △BAP31/GST fusion protein.In our study,high purification △BAP31/GST fusion protein is obtained through the E.coil expression system.

A highly specific and sensitive microplate chemiluminescent enzyme immunoassay (CLEIA) was established and validated for the detection of staphylococcal enterotoxin B (SEB). A pair of monoclonal antibodies (mAbs) that recognizes different epitopes of SEB was selected from 20 SEB-specific mAbs, and the experimental conditions were examined and optimized for the development of the CLEIA. This method exhibited high performance with a dynamic range of 0.01-5 ng/mL, and the measured limit of detection (LOD) was 0.01 ng/mL. Intra- and interassay coefficient variations were all lower than 13% at three concentrations (0.2, 0.4, and 2 ng/mL). For specificity studies, when this method was applied to test staphylococcal enterotoxins A, C1, and D, no cross-reactivity was observed. It has been successfully applied to the analysis of SEB in a variety of environmental, biological and humoral matrices such as sewage, tap water, river water, roast beef, peanut butter, cured ham, 10% nonfat dry milk, milk, orange juice, and human urine and serum. The aim of this article is to show that the highly sensitive, specific, and simple microplate CLEIA, based on a pair of highly specific monoclonal antibodies, has potential applications for quantifying SEB in public health and military reconnaissance.

目的:构建重组表达载体P-BAP31和P-L-BAP31基因疫苗并接种小鼠,评价其诱导的体液和细胞免疫应答。方法:利用分子克隆技术,构建重组质粒P-BAP31和P-L-BAP31,同时构建P-BAP31/EGFP和P-L-BAP31/EGFP重组质粒,并将其以脂质体瞬时转染HeLa细胞,通过荧光显微镜观察目的蛋白的表达;将重组质粒P-BAP31和P-L-BAP31免疫C57BL/6小鼠,采用间接ELISA检测免疫小鼠血清中特异性抗体效价;通过酶联免疫斑点实验(ELISPOT)检测免疫脾细胞在受到BAP31抗原刺激后产生IFN-γ和IL-4细胞因子的频率;通过乳酸脱氢酶(LDH释放)试验检测小鼠特异性细胞毒性T淋巴细胞(CTL)的应答。结果:酶切及测序结果表明,成功构建了针对BAP31肿瘤抗原的P-BAP31和P-L-BAP31基因疫苗。通过荧光显微镜观察重组质粒转染的HeLa细胞,可见EGFP报告基因表达的绿色荧光。ELISA检测免疫小鼠血清中特异性抗体效价发现,带LAMP组明显高于不带LAMP组(P<0.05),且两者均高于空质粒(P-LAMP)及正常对照(N)组(P<0.01)。ELISPOT检测脾细胞分泌细胞因子IFN-γ和IL-4的频率,带LAMP组与其他组相比显著提高(P<0.01)。LDH释放试验显示,带LAMP组的特异性CTL活性高于不带LAMP组,且均高于对照组(P<0.01)。结论:构建了针对肿瘤抗原BAP31的全长基因疫苗,以其免疫C57BL/6小鼠可诱导特异性体液和细胞免疫反应,其中,P-L-BAP31基因疫苗各项免疫反应均优于P-BAP31。

BAP31 is an evolutionarily conserved polytopic integral protein of the endoplasmic reticulum (ER) membrane implicated in regulating the export of selected membrane proteins from the ER to downstream compartments of the secretory pathway. BAP31 interacts with mIgD, cellubrevin, major histocompatibility complex class I, and BCL-2/BCL-X(L) and plays an important role in regulating the egress of these proteins and in apoptosis. Although BAP31 RNA is ubiquitous, the protein's anatomic localization in rat tissues has not been determined. This is partially because production of high affinity antibodies, especially monoclonal antibodies (MAbs) suitable for immunohistochemical staining, has lagged. To gain further insight into its possible functions, we generated a novel MAb specific for rat BAP31 in immunocytochemistry and immunohistochemistry and localized BAP31 in some rat tissues. Immunoreactivity of BAP31 was prominent in fundic glands, colon, pancreatic acinuses, and liver but not in skeleton muscle and lung. Thus, successful production of rat BAP31 monoclonal antibodies provides a new powerful tool for investigation of BAP31 function in the rat model.

AIM To observe the changes of the number of NKT cells in spleens and livers of induced model of experimental autoimmune encephalomyelitis (EAE), and to study the role NKT cells play in the immunoregulation of EAE. METHODS C57BL/6 mice were immunized with MOG (35-55) peptide and received clinical evaluation daily. The mice were sacrificed at the fastigium and the splenic and hepatic lymphocytes were isolated. The changes of NKT cells in normal and EAE C57BL/6 mice were detected by flow cytometry. RESULTS The percent of NKT cells in lymphocytes of different organs of EAE model were greater decreased than in that of normal mice. The percent of NKT cells in splenic lymphocytes of normal mice was 2.22+/-0.14, while that in EAE mice was 1.94+/-0.07 (P<0.05). The percent of NKT cells in hepatic lymphocytes of normal mice was 5.52+/-2.17, while that in EAE mice was 2.67+/-1.41 (P<0.05). CONCLUSION The proliferation of splenic and hepatic NKT cells in C57BL/6 mice are inhibited in EAE model, which may indicate that the immune function conducted by NKT cell is down regulated in EAE mice.