人免疫球蛋白G的原子力显微镜观察

应用原子力显微镜（ＡｔｏｍｉｃｆｏｒｃｅｍｉｃｒｏｓｓｃｏｐｅＡＦＭ）观察不同浓度人ＩｇＧ样品，当ＩｇＧ浓度为２０μｇ／ｍｌ时，ＩｇＧ分子不易在基底表面分散，大部分ＩｇＧ聚集而形成多聚体，当浓度为０．２５μｇ／ｍｌ或０．１μｇ／ｍｌ时，ＩｇＧ分子基本上能较好的分散，可见ＩｇＧ呈“三个球形”结构。当浓度为０．０２μｇ／ｍｌ时，可看到单个ＩｇＧ分子的球形结构，大小为１６．５５～１８．５ｎｍ。     

半巢式 PCR 联合 Southern 杂交检测白血病免疫球蛋白重链(IgH)基因重排

应用ＰＣＲ方法检测免疫球蛋白重链（ＩｇＨ）基因重排可进行白血病微小残留病（ＭＲＤ）研究，应用半巢式ＰＣＲ法检测白血病患者ＩｇＨＣＤＲ－Ⅲ片段，７３．５％（２５／３４）急性淋巴细胞白血病（ＡＬＬ，简称急淋），７５％（３／４）慢性淋巴细胞白血病（ＣＬＬ，简称慢淋），１５．２％（５／３３）急性非淋巴细胞白血病（ＡＮＬＬ，简称急非淋）和０％（０／１９）慢性粒细胞白血病（简称慢粒）存在ＩｇＨ基因重排，所有阳性病例都通过Ｓｏｕｔｈｅｒｎ杂交加以证实，结果显示①半巢式ＰＣＲ较一次ＰＣＲ不仅灵敏度提高，而且在一定程度上能降低假阴性率。②ＩｇＨ重排并不只局限于Ｂ淋巴细胞白血病，还可出现于部分急非淋病例，其机理有待于进一步探讨。     

特异性IgG4在蠕虫病诊断及疗效考核中的价值

IgG4是IgG的亚类之一。在血吸虫等蠕虫感染过程中特异性IgG4在血清中含量可显著增加,有效治疗后则下降较快,因此具有潜在的诊断和疗效考核作用。本文针对特异性IgG4在血吸虫、囊虫、丝虫等蠕虫病诊断方面的研究进行综述。     

IMMUNE RESPONSES IN AUTOIMMUNE HEPATITIS: EFFECT OF PREDNISONE AND AZATHIOPRINE TREATMENT: CASE REPORT

The role of the immune response in autoimmune hepatitis has not been studied before and after prednisone and azathioprine treatment. Distributions of blood lymphocytes (CD4+, CD8+, CD19+, CD23+, CD16/56+), levels of serum immunoglobulins (IgM, IgG, IgE, IgA) and cytokines (IFN-γ, IL-4, IL-12, TNFα ) were studied in a child (f/14 y/o) with autoimmune hepatitis before and after prednisone (20 mg/d) and azathioprine (50 mg/d) treatment (nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, flow cytometry, cytokine ELISA). Patient was studied for 0-2.5 yrs; treatment was initiated 12 weeks post diagnosis. Numbers of CD4+ T cells increased (50%), while CD19+ and CD23+ cells decreased (>50%) post treatment; other lymphocyte subsets were unaffected by treatment. Serum IgG and IgE levels decreased (>50%) after treatment; serum IgM and IgA were within normal range and were not affected by treatment High levels of IFN-γ (5-23 pg/ml) were initially detected in serum, which decreased after treatment (<0.1 pg/ml). Furthermore, low levels of IL-4 (0.2 pg/mL) were detected before treatment, which were not detected after treatment (<0.1 pg/ml). In contrast, before treatment, IL-12 and TNFα were not detected in serum; however after treatment the levels of IL-12 and TNFα dramatically increased. Prednisone and azathioprine treatment decreased total serum IgG, IgE, IFN-γ and IL-4 levels, and blood CD19+ and CD23+ cells; however serum IL-12, TNFα and blood CD4+ T cells increased with treatment. Understanding immunomodulation in autoimmune hepatitis will provide better insight and mechanisms of this disease and may tailor more effective therapeutic intervention.     

用CL法和微量MPO法观察人参皂甙对人PMN吞噬杀菌作用

采用化学发光（ＣＬ）法和髓过氧化物酶（ＭＰＯ）微量测定法检测了人参茎叶皂甙对人多形核白细胞（ＰＭＮ）吞噬金黄色葡萄球菌的影响。实验结果表明，人参茎叶皂甙作用过的ＰＭＮ－ＣＬ反应（ＣＬ３０和Ｐｅａｋ）和ＭＰＯ释放水平均增强，对ＣＬ３０与ＣＦＵ、ＭＰＯ与ＣＦＵ行直线相关分析均呈密切相关（ｒ＝０．９５８，ｐ＜０．００１；ｒ＝０．８７５，ｐ＜０．０１）。本实验提示，ＰＭＮ－ＣＬ法和ＭＰＯ微量测定法是研究药物对ＰＭＮ吞噬杀菌功能影响的客观、定量和快速的方法。     

热塑性聚氨酯/聚乙二醇/纳米铜复合材料的制备与表征

选择热塑性聚氨酯(polyurethane,PU)为基体材料,以聚乙二醇(polyethylene glycol,PEG)对基体材料进行亲水改性,用纳米铜颗粒代替铜丝或铜管等块体铜,采用机械共混的方法制备了一种全新的宫内节育器(IUD)材料——聚氨酯/聚乙二醇/纳米铜复合材料,并采用红外光谱(FTIR)分析、X射线衍射(XRD)分析、扫描电镜分析(SEM)、力学性能测试、热稳定能测试和吸水性能测试等表征方法对复合材料的结构和性能进行了表征。结果表明,纳米铜粉在基体中分散比较均匀,聚氨酯/聚乙二醇/纳米铜复合材料的拉伸强度、撕裂强度随着聚乙二醇含量的增加而下降,断裂伸长率、热稳定性和吸水性能随着聚乙二醇含量的增加而提高,吸水性能的改善为下一步复合材料铜离子的可控释放研究工作提供了很好的基础。     

OCCURRENCE,PROPERTIES, AND FUNCTION OF ASYMMETRIC IgG MOLECULES ISOLATED FROM NON-IMMUNE SERA

We have previously demonstrated that 10–20% of the IgG isolated from non-immune sera is asymmetrically glycosylated, in such a way that it fails to trigger immune effector mechanisms. As a result, a major portion of the non-immune asymmetric IgG molecules of the host could be self-specific, acting as auto-protective antibodies. In order to test this hypothesis, we investigated whether asymmetric IgG molecules are capable of recognizing self-antigens. About 40% of F(ab′)₂ fragment from normal rat IgG was able to react specifically with autologous rat cells. Moreover, upon being purified from normal rat sera, 78% of the asymmetric IgG sub-population showed self-reactivity. We demonstrated that about 14% of rat asymmetric IgG-F(ab′)₂ fragments was able to react with bacteria isolated from the intestine of uninfected rats. Lastly, in order to test whether there is a correlation between the decline of immune responses during ageing and asymmetric antibody production, we assayed IgG isolated from sera of young and old rats. There was an increase in the asymmetric:symmetric IgG ratio with ageing. We therefore suggest that asymmetric antibodies may exert a beneficial action by protecting self-antigens as well as normal intestinal flora from a deleterious immune response.     

应用Protein G纯化细胞培养上清中的大鼠单抗

用ｍｉｎｉＰｅｒｍ系统旋转连续培养大鼠杂交瘤细胞株,获得大量含抗鼠ＩＬ－４（ＩｇＧ１）和ＩＬ－５（ＩｇＧ１和ＩｇＧ２ａ）抗体的培养上清;经ＰＭ１０膜超滤,５０％饱和硫酸铵沉淀及链球菌Ｃ蛋白－琼脂糖亲和层析纯化ＩｇＧ单抗。结果每升杂交瘤细胞２上清得到大鼠,ＩｇＧ２２－３６ｍｇ;１次可纯化抗体２０ｍｇ;经过ＳＤＳ－ＰＡＧＥ及ＥＬＩＳＡ鉴定,所得到的抗体纯度高,活性保持良好。     

神经细胞及其上清液对小鼠腹腔巨噬细胞的作用—细胞化学及MTT定量研究

本研究采用细胞化学和MTT定量的方法,观察鸡胚前脑神经细胞及其上清液(S_1)对小鼠腹腔巨噬细胞的作用。发现神经细胞及S_1可使培养的巨噬细胞酸性磷酸酶(AcP_(ase))和三磷酸腺苷酶(ATP_(ase))含量增加。MTT定量的结果表明在神经细胞及S_1作用下,巨噬细胞OD值比对照组明显高。S_1经各种处理后,进一步观察对巨噬细胞的影响,发现影响巨噬细胞活性的物质,很可能是一种膜糖蛋白。     

免疫吸附技术的临床应用体会

目的　为了评价蛋白A免疫吸附和低密度脂蛋白免疫吸附对免疫球蛋白(主要为IgG)和低密度脂蛋白(LDL)的吸附作用及临床效果。方法　使用瑞典Excorim公司的Citem10吸附-洗脱环路系统,对7例自身免疫性疾病和3例高脂血症患者进行了16例次免疫吸附治疗,并将治疗前后血浆免疫球蛋白或血脂进行对比分析。结果　吸附治疗后血浆IgG、IgA、IgM均明显低于吸附前的水平(P<0.001),LDL、Lp(a)、甘油三脂、胆固醇分别下降了70.9%、53.3%、19.7%、53.0%,而高密度脂蛋白(HDL)较吸附前增高51.3%。结论　蛋白A免疫吸附和低密度脂蛋白免疫吸附可以选择性地、有效地清除血浆IgG及LDL,缓解临床症状,且未见明显副作用。     

多克隆免疫激发剂LPS免疫激发大鼠引起下丘脑和脑干核团c—fos表达

为了解参与机体免疫调控过程的中枢神经核团，对雄性ＳＤ大鼠侧脑室及腹腔注射多克隆免疫激发剂ＬＰＳ，应用即刻早期表达基因ｃ－ｆｏｓ免疫组织２化学法，观察大鼠服内Ｆｏｓ表达。侧脑室注射ＬＰＳ１０ｎｇ，３ｈ后见下丘脑室旁核内侧部对应ＣＲＦ神经元分布区有密集的Ｆｏｓ阳性染色；外侧及腹侧部相当于ＯＸＴ细胞所在部位的脑干，脊髓投射区有Ｆｏｓ表达细胞；前大细胞亚核亦有Ｆｏｓ染色细胞。     

肝细胞生长因子对肺癌细胞运动和侵袭的影响及原癌基因C-met的表达

肝细胞生长因子（ＨＧＦ）可调节许多细胞的运动和生长，检测了ＨＧＦ对５种人类非小细胞肺癌细胞运动和侵袭的作用，有３种细胞的运动和侵袭力在ＨＧＦ刺激后增加。用ＲＮＡ印迹检测ＨＧＦ受体——原癌基因Ｃ－ｍｅｔ产物，可证实这几种细胞均有Ｃ－ｍｅｔｍＲＮＡ存在。细胞蛋白印迹检测进一步证实了ＨＧＦ受体的存在。这些结果表明ＨＧＦ可能在人类非小细胞肺癌的侵袭和转移方面扮演重要角色。     

链球菌脂磷壁酸的提取、生物学特性及其抗肿瘤活性的初步研究

用冷酚抽提法从A群溶血性链球菌中提取了脂磷壁酸(Lipoteichoic acid LTA),经紫外扫描、凝胶过滤对提取物的纯度进行了分析。经红外光谱分析同文献报道相似。间接血球凝集试验结果显示LTA能自主粘附于经双醛法固定的羊红血球,并能同抗LTA抗体或抗A群链球菌抗体结合而使血球凝集。LTA注射小鼠腹腔,能使其脾重显著增加。抗肿瘤效力实验证明LTA能够抑制艾氏癌细胞在小鼠体内(皮下、腹腔)的生长。接种2×10~5个细胞于小鼠皮下后连续7天注射含10μg磷的LTA其抑瘤率为81.9％,与对照组相比(P<0.01)呈显著性差异。当对腹水型动物注射含0.25μg和1μg的LTA时其生命延长率分别为54％和44％。并进一步证明LTA抗肿瘤的机制之一是诱导肿瘤坏死因子TNF的产生。     

Probing the weak interaction of proteins with neutral and zwitterionic antifouling polymers

Protein–polymer interactions are of great interest in a wide range of scientific and technological applications. Neutral poly(ethylene glycol) (PEG) and zwitterionic poly(sulfobetaine methacrylate) (pSBMA) are two well-known nonfouling materials that exhibit strong surface resistance to proteins. However, it still remains unclear or unexplored how PEG and pSBMA interact with proteins in solution. In this work, we examine the interactions between two model proteins (bovine serum albumin and lysozyme) and two typical antifouling polymers of PEG and pSBMA in aqueous solution using fluorescence spectroscopy, atomic force microscopy and nuclear magnetic resonance. The effect of protein:polymer mass ratios on the interactions is also examined. Collective data clearly demonstrate the existence of weak hydrophobic interactions between PEG and proteins, while there are no detectable interactions between pSBMA and proteins. The elimination of protein interaction with pSBMA could be due to an enhanced surface hydration of zwitterionic groups in pSBMA. New evidence is given to demonstrate the interactions between PEG and proteins, which are often neglected in the literature because the PEG–protein interactions are weak and reversible, as well as the structural change caused by hydrophobic interaction. This work provides a better fundamental understanding of the intrinsic structure–activity relationship of polymers underlying polymer–protein interactions, which are important for designing new biomaterials for biosensor, medical diagnostics and drug delivery applications.     

儿童缺氧缺血性脑病的影像学和脑电图检查结果的对照及意义

目的：观察影像学和脑电图对儿童缺氧缺血性脑病的应用对照及意义。方法：对65例缺氧缺血性患儿作影像学和脑电图检查。结果：脑电图正常6例（9．23％），轻度异常13例（22．03％），中度异常37例（61．03％），重度异常9例（16．94％），脑电图异常率为90．77％。脑电图异常表现为弥漫性慢波增多和局限性癎性放电。影像学异常44例，异常率为67．69％。结论：影像学和脑电图检查对儿童缺氧缺血性脑病的诊断及治疗有一定的帮助。     

检测类风湿因子ELISA法的建立及其应用

本文建立了测定类风湿因子（ＲＦ）总量的双抗原夹心法和测定ＩｇＧ－ＲＦ的间接法。用于临床检测表明，１００名献血员中有３例ＲＦ总量测定为阳性。９７例类风关患者中７２例阳性（阳性率７５．３％）。１３２例健康老年人中５人阳性（阳性率３．８％）。ＩｇＧ－ＲＦ测定结果：１２０例正常人和１３２例健康老年人均为阴性，９７例类风关病人中有７例阳性（阳性率７．２％）。     

Characterization of PEG–iron oxide hydrogel nanocomposites for dual hyperthermia and paclitaxel delivery

Hyperthermia, the heating of tissue from 41 to 45?°C, has been shown to improve the efficacy of cancer therapy when used in conjunction with irradiation and/or chemotherapy. In this work, hydrogel nanocomposites have been developed that can control the delivery of both heat and a chemotherapeutic agent (e.g. paclitaxel). The nanocomposites studied involve a stealth, poly(ethylene glycol) (PEG)-based system comprised of PEG (n?=?1000) methyl ether methacrylate and PEG (n?=?400) dimethacrylate with iron oxide nanoparticles physically entrapped within the hydrogel matrices. The capability of the hydrogel nanocomposites to be heated in an alternating magnetic field was demonstrated. The heating of the hydrogel systems was dependent on the crosslinking of the hydrogel network where hydrogels with lower swelling ratios were found to heat to a greater extent than those with higher ratios. In addition, paclitaxel was shown to exhibit non-Fickian release from the hydrogel systems, with the amount of drug released dependent on the hydrogel network structure. Three cell lines: M059K (glioblastoma), MDA MB 231 (breast carcinoma), and A549 (lung adenocarcinoma) were exposed to paclitaxel only, hyperthermia only, and both paclitaxel and hyperthermia to determine if a synergistic cytotoxic effect was possible for these cell lines. The efficacy of paclitaxel was greater with hyperthermia for the A549 cells; however, the M059K and MDA MB 231 did not show the same response.     

New Insights on the Self‐Organization of PEG/α‐Cyclodextrin‐Based Polyrotaxanes in Solution: An Experimental and Theoretical Study

Size exclusion chromatography coupled with light scattering (SEC/MALS), dynamic light scattering (DLS), steady‐state and time‐resolved fluorescence, as well as molecular dynamics (MD) simulations are used to study the behavior of several poly(ethylene glycol) (PEG)/α‐cyclodextrins (αCDs) polyrotaxanes (PRs) in solution. The number of CD units in any of the PRs studied is always smaller than that required to saturate the PEG chains. These PRs seem to aggregate in dimethyl sulfoxide (DMSO) solution. The presence of hairpins in the non‐saturated PRs contributes to diminishing their expected large dimensions. Intra‐ and inter­molecular interactions and forces responsible for hairpins and aggregation are investigated.

     

在体和离体的鸡背根节神经细胞5-羟色胺的免疫组织化学研究

用PAP免疫组织化学方法研究了鸡胚和雏鸡背根节内5-HT神经细胞的形态与胚胎发育,并在离体细胞培养条件下研究了靶周围组织(皮肤)对其胚胎发育的影响。在鸡腰骶部背根节,5-HT免疫反应阳性细胞最先出现于鸡胚13d(E_(13)),占0.4%;孵出后2d(AH_2)占6.2%。阳性细胞主要为大的A类细胞和极少的小B类细胞。免疫反应阳性的周围神经末梢位于皮肤和跟腱中。取自E_9鸡胚的背根节细胞培养7d后有部分出现5-HT阳性免疫反应,取自E_6鸡胚的背根节细胞培养10d仍为阴性反应;然而,当取自E_6鸡胚的背根节细胞与皮肤组织联合培养10d后,则出现5-HT阳性免疫反应细胞。本文还对脊髓5-HT细胞的出现进行了讨论。     

人肿瘤细胞与正常细胞内微管的PAP免疫酶细胞化学观察

我们利用兔抗微管蛋白抗体和兔抗辣根过氧化物酶(HRP)抗血清制备的HRP—抗HRP(PAP)复合物,建立了微管的PAP免疫酶细胞化学方法。应用此法观察到人食管癌ECa 109、胃癌SGC 7901,乳腺癌MCF 7和成骨肉瘤OS 732细胞间期胞质微管减少或消失,只有大量弥散分布的微管蛋白棕色反应产物,在微管组织中心(MTOC)附近十分密集,而正常成纤维细胞和胎儿胃粘膜上皮细胞间期,都有发达的胞质微管结构(CMTC)。在分裂期,这些肿瘤细胞都显示纺锤体微管,与正常细胞比较无明显差异。本研究应用PAP方法进一步证明,以前用免疫荧光细胞化学方法观察到的人肿瘤细胞间期胞质微管缺陷的特征。除去低温(4℃)或秋水仙酰胺处理后,解聚的CMTC又可恢复,表明本方法与免疫荧光染色法,同样具有很高的特异性。本工作在细胞固定及免疫反应的某些步骤上有所改进。