Effect of illumination of nitrate and phosphate removal by coimmobilized Chlorella pyrenoidosa and activated sludge

Chlorella pyrenoidosa and activated sludge were co-immobilized with simplifying modified PVA-sulfate method. Effects of light intensity and light:dark ratio on the growth of co-immobilized algae cells and removal of nitrate and phosphate were studied. The results indicate that the growth rate of co-immobilized algae cells reduces when light intensity is decreased from 4000 Lux to 1000 Lux, while optimal light:dark ratio for algal growth is 16:8. The influence of illumination on nitrate removal is so weak that the removal percentage can reach 90 approximately 100% within 12 approximately 24 h during the experimental periods in spite of changing illumination conditions. On the other hand, phosphate removal efficiency reduces when light intensity or light:dark ratio is decreased. The highest phosphate removal percentage is 99.6% under the circumstances of 4000 Lux and full-time illumination in our experiment, while the average phosphate removal is about 78%. The change of pH value in water samples is also observed. When water sample is treated by the co-immobilized system, pH value increases in light and decreases in dark. Microorganisms' physiological action is considered as the main mechanism that leads to the change of pH value.     

The ability of heat-killed Mycobacterium vaccae to stimulate a cytotoxic T-cell response to an unrelated protein is associated with a 65 kilodalton heat-shock protein

Exogenous antigens are generally presented by Class II major histocompatibility (MHC) molecules. When administered with an adjuvant, however, they are capable of inducing a CD8+ T-cell response where antigen recognition is associated with Class I MHC. Accordingly, immunization with soluble ovalbumin (OVA) alone does not activate CD8+ cytotoxic T cells (CTL) but when given in complete Freund's adjuvant (CFA), or in formulations of a number of novel adjuvants, an OVA-specific CD8+ CTL response can be detected. We show in this report that immunization with soluble OVA mixed with heat-killed Mycobacterium vaccae, but not with other common pathogenic and saprophytic mycobacteria, can activate OVA-specific CD8+ CTL. An OVA-specific CTL response is detected when mice are immunized by either the intraperitoneal or intranasal route and their spleen cells are re-stimulated in vitro. Adjuvant activity of heat-killed M. vaccae is present in M. vaccae culture filtrate, in soluble protein components of whole M. vaccae and in the 65 kDa heat-shock protein (hsp) of M. vaccae. Mycobacterium vaccae has previously been shown to have no adverse side-effects in humans. The current results suggest that M. vaccae may be useful as an adjuvant for vaccines and other immunotherapies where CD8+ CTL responses to exogenous proteins are crucial.     

Nucleolar localization of non-structural protein 3b, a protein specifically encoded by the severe acute respiratory syndrome coronavirus

The open reading frame 3 (ORF3) of the severe acute respiratory syndrome coronavirus (SARS-CoV) genome encodes a predicted 154-amino acid protein, which lacks similarities to any known protein, and is named 3b. In this study, it was shown that 3b protein was predominately localized to nucleus with EGFP tag at its N- or C-terminus. The localization patterns were similar in different transfected cells. Immuno-fluorescence assay revealed that 3b protein was co-localized well with C23 in nucleolus. C23, B23 and fibrillarin all are important nucleolar proteins, which localize in the region of the nucleolus. Co-transfection of p3b-EGFP with pC23-DsRed, pB23-DsRed and pfibrillarin-DsRed further confirmed 3b's nucleolus localization. With construction of serial truncated mutants of 3b, a region (residues 134-154 aa) responsible for nucleolar localization was determinated in 3b protein. These results provide a new insight for further functional studies of SARS-CoV 3b protein.     

非悬滴开放式培养法在鸡胚背根节体外培养中的应用

本研究针对悬滴培养法在操作和应用上存在的问题和局限性，改用操作简便、适用范围广的非悬滴开放式培养法培养鸡胚背根节。将数个鸡胚背根节按一定间隔种植在内置生长基质盖玻片的35mm培养皿中，加人适量培养液，置于CO2。孵箱中进行培养。结果显示.从培养24h至60h各时期，培养皿中背根节生长状况均良好，神经突起明显增长，表明用非悬滴开放式培养法培养鸡胚背根节是可行且可靠的。     

IV型II类反式激活因子基因新变异体的克隆、鉴定和功能测定

为获取有功能的IV型II类反式激活因子基因 (CIITA IV ) ,诱导肿瘤细胞表达MHCII类分子 ,从IFN γ刺激的THP 1细胞中以RT PCR获得CIITA IV ,将其连接到pGEMT easy载体。对所构建的pcDNA3 1 CIITA IV型表达载体进行反复测序后发现 ,所获得的CIITA IV基因存在结构变异 ,在 2 87位插入了 3个核苷酸TAG ,使 2 86 2 88位的AAG改变成为ATAGAG(2 86 2 90 ) ,并引起其他 8个座位核苷酸 (及推导的氨基酸残基 )发生改变。将表达载体转入原先不表达MHCII类分子的HeLa细胞中 ,检测到所获得的IV型CIITA变异体具有诱导人II类分子HLA DR表达的能力。空载体和CIITA IV基因导入的HeLa细胞中 ,DR阳性细胞百分率分别为 0 0 1 %和 37 6 4 %。该基因已从GenBank得到登录号 ,表明这是一个具有诱导HLA DR分子表达功能的IV型CIITA新基因。     

周围神经干自然分束部位血管形态及临床意义

应用微血管灌注透明法观察研究5具成人周围神经干标本.重点观察神经自然分束部位微血管形态和分支分布规律.神经干内有非常完兽的血供系统.手术中循神经自然分束进行追踪分离,不会对神经血供产生明显影响.但在张力下牵拉缝合神经,将对神经干血供产生严重影响.神经束膜缝合比神经外膜缝合更强调在无张力下进行.     

Activation of the parapyramidal region in the ventral medulla stimulates gastric acid secretion through vagal pathways in rats

Neurons synthesizing thyrotropin-releasing hormone, substance P and serotonin in the medullary caudal raphe nuclei project to the dorsal vagal complex and play a role in the central vagal regulation of gastric function. Neurons in the parapyramidal region in the ventral medulla share similar biochemical coding and projections as those in the caudal raphe nuclei. The role of the parapyramidal region in the autonomic regulation of gastric acid secretion was investigated in urethane-anesthetized rats. Unilateral microinjection of kainate into the parapyramidal region at 10, 15 and 20 ng induced a dose-related stimulation of gastric acid secretion (net increases: 22.2+/-11.2, 40.5+/-8.5 and 89.8+/-19.4 micromol/60 min, respectively), while injection of vehicle had no effect (net change: -0.1+/-1.4 micromol/60 min). Time-course studies showed a nine-fold peak increase over basal at 30 min after parapyramidal injection of kainate (20 ng) and acid secretion returned to basal level at 70 min. Microinjections of kainate (15-20 ng) outside the parapyramidal region or into the parapyramidal region in vagotomized rats had no effect. Exposure to cold (4 degrees C) for 2 h, which is known to induce vagally mediated gastric secretory and motor responses through medullary thyrotropin-releasing hormone pathways, increased the number of Fos-positive cells in the caudal, middle and rostral parts of the parapyramidal region to 4.3+/-0.4, 9.4+/-0.9 and 18.4+/-1.6/section, respectively, compared with 0.1+/-0. 1, 0.1+/-0.0 and 0.7+/-0.6/section, respectively, in rats maintained at room temperature. Most of the Fos-labeled cells co-expressed pro-thyrotropin-releasing hormone messenger RNA signal and/or were serotonin immunoreactive. These data show that chemical activation of neurons in the parapyramidal region results in a vagal-dependent stimulation of gastric acid secretion and that acute cold exposure activates parapyramidal neurons containing pro-thyrotropin-releasing hormone and/or serotonin, suggesting a potential role of the parapyramidal region, in addition to the caudal raphe nuclei, as medullary sites involved in the vagal regulation of gastric function.     

A tctex1-Ca2+ channel complex for selective surface expression of Ca2+ channels in neurons

Voltage-gated Ca(2+) channels (VGCCs) are important in regulating a variety of cellular functions in neurons. It remains poorly understood how VGCCs with different functions are sorted within neurons. Here we show that the t-complex testis-expressed 1 (tctex1) protein, a light-chain subunit of the dynein motor complex, interacts directly and selectively with N- and P/Q-type Ca(2+) channels, but not L-type Ca(2+) channels. The interaction is insensitive to Ca(2+). Overexpression in hippocampal neurons of a channel fragment containing the binding domain for tctex1 significantly decreases the surface expression of endogenous N- and P/Q-type Ca(2+) channels but not L-type Ca(2+) channels, as determined by immunostaining. Furthermore, disruption of the tctex1-Ca(2+) channel interaction significantly reduces the Ca(2+) current density in hippocampal neurons. These results underscore the importance of the specific tctex1-channel interaction in determining sorting and trafficking of neuronal Ca(2+) channels with different functionalities.