FINE STRUCTURE OF NEUROBLASTOMA—A CASE REPORT—

Ultrastructural observation was carried out on neuroblastoma cells in a 4-year-old girl. The nuclei of these neoplastic cells were irregular with occasional cytoplasmic invaginations, and proportionally large with diffusely dispersed chromatin substance. Rare mitotic figures were also present. The cell-to-cell interfaces were relatively simple with only finely wavy membranes but without interdigitations. A few desmosomes were, however, apparent. The cytoplasmic pseudopodes were frequently observed in a limited area of the cell surface and nearly free of organelles. The main findings of the intracytoplasmic organelles consisted of some mitochondrial changes, poor development of Golgi bodies, occasional formation of focal cytoplasmic degradation, and intracytoplasmic Abrillar aggregates. Numerous large round to oval dense bodies encircled by a double membrane were highly suggestive of lipid bodies. The central area of rosette-like cell aggregates was noted to be composed of cytoplasmic processes of neoplastic cells.     

人干扰素基因植物表达载体的构建及其在人参愈伤组织细胞中的表达

目的：利用组织培养人参愈伤组织细胞表达人干扰素，探讨用植物细胞表达人类基因的可行性。方法：用PCR从pBV889扩增人干扰素α2b（hIFN-α2b）编码基因，将其克隆于pMD18-T载体和pBI121，进而构建植物细胞表达载体pBIFN。用冻融法将pBIFN导入根瘤农杆菌LBA4404，经卡那霉素和利福霉素筛选后利用农杆菌介导的转化法将hIFN-α2b基因导入人参愈伤组织细胞。经G418筛选，形成阳性细胞。用PCR、RT—PCR、Western blot和WISH／VSV系统检测转基因组织。结果：经PCR检测表明hIFN-α2b基因已成功整合到人参愈伤组织细胞基因组中。RT-PCR证明存在hIFN-α2b基因的转录产物。Western blot分析表明转基因人参愈伤组织细胞能有效表达hIFN-α2b蛋白。WISH／VSV系统检测分析表明表达的hIFN-α2b具有生物学活性。结论：本研究利用转基因人参愈伤组织细胞成功地表达了人干扰素，为进一步提高口服干扰素的疗效打下基础。     

极低频脉冲电磁场对新生大鼠成骨细胞的影响

目的：观察极低频脉冲电磁场（pulsed electromagnetic fields，PEMF）对体外培养成骨细胞增殖、分化、体外矿化的影响。方法：采用频率为15Hz、强度为5mT、占空比为15％的PEMF作用于成骨细胞，检测成骨细胞的增殖、碱性磷酸酶（alkaline phosphatase，ALP）活性以及体外矿化指标。结果与结论：PEMF显著促进成骨细胞增殖和体外矿化，抑制ALP活性作用。     

Heat shock up-regulates TLR9 expression in human B cells through activation of ERK and NF-kappaB signal pathways

Toll-like receptors (TLRs) play a critical role in innate immunity and TLR9 is essential for CpG ODN signaling. As "dangerous signal", heat shock may regulate immune response. However, little is known about TLRs expression and signaling after heat shock. In this study, we investigated regulation of TLR9 expression and function in human B cell line RPMI8226 by heat shock. We demonstrated that TLR9 expression was up-regulated remarkably following heat shock. Coincidently, CpG ODN stimulation significantly increased IL-6 production and up-regulated expressions of MHC I, MHC II and CD86 by heat-shocked B cells. Heat shock activated ERK and NF-kappaB signal pathways, and pretreatment of B cells with specific inhibitors of ERK or NF-kappaB signal pathways inhibited heat shock-induced up-regulation of TLR9 expression. These results demonstrated that heat shock promotes TLR9 expression and signaling through activation of ERK and NF-kappaB signal pathways in B cells, suggesting that heat shock might modulate host immune response by regulating TLR expression.     

探讨BCG初次免疫结核杆菌DNA疫苗加强免疫方案的效应

目的:探讨BCG初次免疫(BCG-prime),结核杆菌共表达DNA疫苗加强免疫(DNA疫苗-boost)的策略对小鼠的免疫效果。方法:将BCG及结核杆菌重组DNA疫苗依次免疫小鼠,通过检测CTL和NK细胞的杀伤活性和特异性淋巴细胞增殖,以及小鼠血清抗体及细胞因子的水平,观测BCG-prime、共表达结核杆菌Ag85A/GM-CSFDNA疫苗boost策略对小鼠的免疫效果。结果:采用prime-boost免疫策略组的小鼠CTL的杀伤活性明显增强、特异性淋巴细胞明显增殖、IFN-γ的水平明显增高,NK细胞杀伤活性与对照组相比也有一定提高,但未超过BCG单独免疫效果。免疫小鼠血清特异性抗体的滴度超过单独DNA疫苗免疫组。结论:在采用BCG-prime-结核杆菌DNA疫苗boost免疫策略后,能增强对小鼠的免疫效应,尤其是Th1型细胞免疫反应增强明显,为进一步在动物体内进行保护性效应试验的研究提供了实验依据。     

Galactosylated chitosan as a synthetic extracellular matrix for hepatocytes attachment

Galactose moiety as the hepatocyte anchorage was covalently coupled with chitosan for the development of synthetic extracellular matrix. Hepatocytes adhesion to galactosylated chitosan (GC)-coated polystyrene (PS) dish became as high as 94.7% after 2 h incubation whereas the hepatocytes adhesion to chitosan-coated PS dish was 69.1%, indication of galactose-specific recognition between GC molecules and asialoglycoprotein receptors of hepatocytes. The DNA synthesis of the hepatocytes adhered to GC-coated dish was increased in the presence of epidermal growth factor (EGF) at low concentration of GC (0.05 microg/ml) whereas the DNA synthesis of the hepatocytes adhered to GC-coated dish was decreased in the presence of EGF at high concentration of GC (5 microg/ml). The spreading shapes of the hepatocytes adhered to the surface in the presence of EGF at low concentration of GC (0.05 microg/ml) were enhanced than in the absence of EGF. The hepatocytes adhered to the surface at high concentration of GC (5 microg/ml) showed round shapes and exhibited many spheroid formation after 24 h in the presence of EGF.     

Galactosylated PVDF membrane promotes hepatocyte attachment and functional maintenance

One of the major challenges in BLAD design is to develop functional substrates suitable for hepatocyte attachment and functional maintenance. In the present study, we designed a poly(vinylidene difluoride) (PVDF) surface coated with galactose-tethered Pluronic polymer. The galactose-derived Pluronic F68 (F68-Gal) was adsorbed on PVDF membrane through hydrophobic-hydrophobic interaction between PVDF and the polypropylene oxide segment in Pluronic. The galactose density on the modified PVDF surface increased with the concentration of the F68-Gal solution, reaching 15.4 nmol galactosyl groups per cm2 when a 1 mg/ml of F68-Gal solution was used. The adsorbed F68-Gal remained relatively stable in culture medium. Rat hepatocytes attachment efficiency on F68-Gal modified PVDF membrane was similar to that on collagen-coated surface. The attached hepatocytes on PVDF/F68-Gal membrane self-assembled into multi-cellular spheroids after 1 day of culture. These attached hepatocytes in spheroids exhibited higher cell functions such as albumin synthesis and P450 1A1 detoxification function compared to unmodified PVDF membrane and collagen-coated surface. These results suggest the potential of this galactose-immobilized PVDF membrane as a suitable substrate for hepatocyte culture.     

太空发育鸡胚的前庭感受器细胞形态学研究

为了探讨太空微重力对鸡胚前庭感觉上皮细胞的形态发育的影响 ,选取在航天飞机 (STS-2 9)发育鸡胚和地面发育鸡胚各两只 ,利用计算机显微测量技术分别测量椭圆囊和球囊的毛细胞、支持细胞核的切面面积、周长、形状系数。太空发育鸡胚的球囊支持细胞核的切面面积、周长显著大于地面组 ,形状系数无差异 ;太空发育鸡胚的椭圆囊支持细胞核的切面面积、周长、形状系数以及椭圆囊和球囊毛细胞的切面面积、周长与地面发育鸡胚相比无明显差异。微重力可能对球囊支持细胞核的体积发育有影响 ,对椭圆囊和球囊的毛细胞以及椭圆囊支持细胞核的形态发育无影响。     

胃肠道癌患者血清中抗癌胚抗原(CEA)抗体的检测及意义

目的：探讨循环中抗癌胚抗原（ＣＥＡ）特异性抗体的情况，评价ＣＥＡ及抗体的联合检测在胃肠道癌诊断中的作用。方法：用放免法检测血清中 ＣＥＡ含量，用酶联免疫吸附法（ＥＬＩＳＡ）检测抗 ＣＥＡ ＩｇＧ抗体，用竞争抑制法检测抗体的特异性。结果：胃肠道癌患者血清 ＣＥＡ含量升高者（≥１５ ｎｇ／ｍｌ）为 ３０．９％（２１／６８），抗 ＣＥＡ ＩｇＧ抗体阳性者为３５．３％（２４／６８），ＣＦＡ及抗 ＣＥＡ抗体的联合检测可使阳性率提高到５４．３％（３７／６８）；胃肠道良性疾病患者（多发性息肉、溃疡、胰腺炎等）血清ＣＥＡ升高者为３．３％（１／３０），抗 ＣＥＡ　ＩｇＧ抗体阳性者为 ３．３％（１／３０）；健康对照组血清 ＣＥＡ升高者为 ０，抗 ＣＥＡ ＩｇＧ抗体阳性者为 ２．５％（１／４０）。结论：胃肠道癌患者血清中抗癌胚抗原（ＣＥＡ）抗体的检出率较高，这些抗体可作为胃肠道癌的一种肿瘤标志物。