IL-13对小鼠哮喘模型气道黏蛋白基因Muc5ac及Bcl-2、Bax表达的作用

目的研究白细胞介素-13(IL-13)处理小鼠支气管哮喘(哮喘)模型前后肺组织黏蛋白基因Muc5ac、凋亡相关蛋白Bcl-2和Bax表达的作用,探讨气道黏液过度分泌的机制.方法45只雄性BALB/c小鼠随机分为对照组、哮喘组和IL-13组,每组15只.用逆转录-聚合酶链反应(RT-PCR)方法和免疫组化法分别检测Muc5acmRNA、Muc5ac蛋白、Bcl-2蛋白以及Bax蛋白在肺组织的表达.结果哮喘组和对照组肺组织Muc5acmRNA分别为(0.1552±0.0057)和(0.0633±0.0013),Muc5ac蛋白分别为(0.8849±0.0257)和(0.1166±0.0064),两组比较差异均有统计学意义(P<0.01);IL-13组肺组织Muc5acmRNA和蛋白分别为(0.2807±0.0027)和(1.6138±0.0483),与哮喘组、对照组比较差异也均有统计学意义(P均<0.01).与对照组Bcl-2蛋白(0.3279±0.0136)、Bax蛋白(1.7284±0.0263)相比,哮喘组分别增加和降低(分别为0.8383±0.0310和0.8987±0.0106),两组差异均有统计学意义(P均<0.01);IL-13处理后可分别促进Bcl-2和Bax蛋白增加和降低(分别为1.6934±0.0229和0.3522±0.0152),其和哮喘组的差异均有统计学意义(P均<0.01);哮喘组和IL-13组小鼠肺组织Muc5acmRNA、蛋白表达与Bcl-2蛋白表达均呈直线正相关(P均<0.05),而与Bax蛋白表达则均呈直线负相关(P均<0.05).结论IL-13是引起哮喘气道黏液过度分泌的重要细胞因子,它可能通过改变Bcl-2和Bax的表达导致了上述病变.     

体外厌氧条件下载铜蒙脱石杀菌效果的研究

载铜蒙脱石的杀菌作用通过脑心浸液肉汤 (BHI)二倍系列稀释法来判定其最小抑菌浓度 (MIC)。释放入肉汤和生理盐水中Cu2 的量通过原子吸收光谱仪测定。杀菌动力学研究采用改良的振荡瓶法。结果表明 :厌氧条件下 ,载铜蒙脱石具有很强的杀菌性能。载铜蒙脱石对放线共生放线杆菌的最小抑菌浓度为 2 .5 6mg/ml;对血液链球菌为 5 .12mg/ml。测得肉汤二倍稀释的载铜蒙脱石释放出的Cu2 量在 2 .39～ 38.6 5 ppm之间 ,在生理盐水中释放的Cu2 量为 1.85～ 15 .82ppm。本试验结果表明 ,蒙脱石无抗菌性能。     

Acidic ATP activates lymphocyte outwardly rectifying chloride channels via a novel pathway

Using whole-cell patch-clamp techniques we found that ATP activated an outwardly rectifying current in Daudi human B lymphoma cells under acidic conditions. The substitution of Cl^– for gluconate^– shifted the reversal potential, while Cl^– channel blockers, 4,4-diisothiocyanostibene-2,2-disulfonic acid (DIDS) and 9-anthracene carboxylic acid (9-AC), blocked the current, indicating that ATP induces this current by activating the outwardly rectifying chloride channel (ORCC). The effect of ATP on ORCC was mimicked by ADP, but not by other P2 receptor agonists such as ATPS (a poorly hydrolyzable analog of ATP), 2,3-O-benzoyl-4-benzoyl-ATP (BzATP), and UTP. The ATP-induced ORCC current was completely blocked by 100 M suramin (a P2 receptor antagonist), and was partially blocked by 100 M pyridoxal-phosphate-6-azophenyl-2,4-disulfonic acid tetrasodium (PPADS), which is another P2 receptor antagonist. Neither inactivation of G proteins nor elimination of extracellular Ca²⁺ affected the ATP-induced current, indicating that G protein-coupled P2Y receptors and Ca²⁺-permeable P2X receptors are not involved. Based on the pharmacological profile and the fact that acidic conditions are required for ATP to activate the ORCC, we suggest that acidic ATP activates the lymphocyte ORCC via a novel pathway, which is not associated with any previously described purinergic receptors.     

Kinetics of viremia and acute liver injury in relation to outcome of neonatal woodchuck hepatitis virus infection

The kinetics of serum viral responses and acute liver injury were studied during neonatal woodchuck hepatitis virus (WHV) infection in relation to the chronic or resolved outcome. The mean concentrations of serum WHV DNA and surface antigen were significantly higher by week 10 post infection in chronic infections compared to resolving infections, and diverged even further by the time of peak viral load development in serum (week 12). After week 12, these viral markers were detected less frequently with time and at lower concentrations in the resolved outcome. In both outcomes, mean serum activities of hepatic enzymes became increased significantly above baseline by weeks 10-12, peaked at week 14, and normalized by weeks 20-22, thus indicating transient acute liver injury. The increasing liver injury responses were comparable between outcomes at week 12, when serum viral load was markedly higher in the developing chronic infections. This suggested a deficiency in early non-cytolytic control of infection in the chronic outcome. At week 14, liver injury was significantly greater in the resolved outcome and associated with higher mean Fas ligand (FasL) and perforin messenger RNAs (mRNAs) in liver compared to the chronic outcome. This indicated greater immune-mediated killing of infected hepatocytes during resolution. Thus, chronicity as an outcome of neonatal WHV infection develops relatively early during the acute phase of infection due to reduced immune-mediated clearance of infected hepatocytes by both cytolytic and non-cytolytic processes.     

Interleukin-10 secreted by B-1 cells modulates the phagocytic activity of murine macrophages in vitro

As demonstrated previously in our laboratory, B-1 cells migrate from the peritoneal cavity of mice and home to a distant site of inflammation to become macrophage-like cells. However, the influence that these cells might have on the kinetics and fate of the inflammatory process is not known. Considering that macrophages are pivotal in the inflammatory reaction, we decided to investigate the possible influence B-1 cells could have on macrophage activities in vitro. Our results show that peritoneal macrophages from Xid mice, a mouse strain deprived of B-1 cells, have higher phagocytic indexes for zymozan particles when compared with macrophages from wild-type mice. Moreover, macrophages from wild-type mice have a lower ability to release nitric oxide and hydrogen peroxide when compared with macrophages from Xid mice. Experiments using cocultures of B-1 cells and macrophages from Xid mice in transwell plates demonstrated that B-1 cells down-regulate macrophage activities. These observations also indicate that this phenomenon is not due to a physical interaction between these two cell populations. As B-1 cells are one of the main sources of interleukin (IL)-10, we demonstrate in this study that adherent peritoneal cells from Xid mice produce significantly less amounts of this cytokine in culture when compared with IL-10 production by cells from wild-type mice. When B-1 cells from IL-10 knock-out mice and macrophages from wild-type mice were cocultured in transwell plates, the phagocytic index of macrophages was not altered demonstrating that B-1 cells can influence the effector functions of macrophages in vitro via IL-10 secretion.     

应用聚合酶链反应技术检测肝组织中的HBV－DNA

采用聚合酶链反应（ＰＣＲ）技术，对４２例肝活切组织石蜡切片中乙型肝炎病毒（ＨＢＶ）ＤＮＡ进行检测，并与乙肝表面抗原（ＨＢｓＡｇ）的免疫组织化学及血清学检测进行比较，ＨＢＶ－ＰＣＲ阳性率为７３．８％，高于组织及血清ＨＢｓＡｇ阳性率（分别为５９．５％和５０．０％）。３例病理形态呈肝炎改变，而血清ＨＢｓＡｇ（─）的肝组织中有２例检出ＨＢＶ－ＤＮＡ，提示ＰＣＲ的高度敏感性和准确性。８３．３％的门脉性肝硬变和８７．５％的肝细胞癌组织中ＨＢＶ－ＰＣＲ呈阳性，进一步证实了上述两病与ＨＢＶ的关系密切。我们还发现肝细胞淤胆患者ＨＢＶ感染率较高，ＨＢＶ－ＤＮＡ及组织ＨＢｓＡｇ阳性比例各为６／９和４／８。     

胎肝细胞质液对重症病毒性肝炎患者外周血TL－CFU及mIL－2R影响的实验观察

采用半固体一步单层琼脂培养法和单克隆荧光抗体技术分别观察重症肝炎外周血ＴＬ－ＣＦＵ和ｍＩＬ－２Ｒ，发现重症肝炎患者ＴＬ－ＣＦＵ（１０４．４±３２．６）及ｍＩＬ－２Ｒ（３５．６±８．６）较正常人明显降低。在培养体系中加胎肝细胞质液后，无论在重症肝炎组还是在正常组均不能明显地提高ＴＬ－ＣＦＵ，说明胎肝细胞质液不含具生物佐的促ＴＬ－ＣＦＵ因子。但对ｍＩＬ－２Ｒ表达的影响，在重症肝炎组病人，只有在ＰＨＡ存在条件下才能促进ｍＩＬ－２Ｒ的表达，说明胎肝细胞质波含有某种（些）物质能协同ＰＨＡ促进重症肝炎患者外周血淋巴细胞ｍＩＬ－２Ｒ表达。     

Role of glomerular prostanoid in control of glomerular filtration rate in rats

Summary It is generally accepted that the main action of glomerular prostanoids (GPs) on glomerular filtration rate (GFR) is to modulate the activity of different vasoconstrictors, specially in states of renal hypoperfusion. However it was also suggested that GPs may directly affect GFR. The present study was focused on this last hypothesis, in different experimental models, in rats.In adriamycin induced acute renal failure, the transient decrease of GFR is associated with higher levels of thromboxane B₂. Later on, when GFR returns to normal, vasodilator prostaglandins synthesis was also increased.In captopril induced renal failure in Na depleted rats (where GPs synthesis remained normal), stimulation of PGE₂ and PGI₂ production by K and NaCl was associated with a significant improvement of GFR. Furthermore, the increase in GFR induce by NaCl was prevented by inhibition of prostaglandin synthesis.Infusion of atrial natriuretic peptide in euvolemic rats induce a marked elevation both of GFR and PGE₂ synthesis. It was abolished by previous administration of prostaglandin synthesis inhibitor.In conclusion, glomerular prostanoids may influence GFR, either directly, or as mediator or modulator of other vasoactive hormones.Abbreviations GPs glomerular prostanoids - PG prostaglandin - TX thromboxane - ANP atrial natriuretic peptide - GFR glomerular filtration rate - PE polyethylene Nachtrag zu den Hauptreferaten des 19. Kongresses der Gesellschaft für Nephrologie in Göttingen (Klin Wochenschr 66/18)     

超抗原金黄色葡萄球菌肠毒素基因B的原核表达

目的构建重组pET-30a-SEB原核表达载体,转化感受态大肠杆菌BL-21(DE3),诱导表达超抗原金黄色葡萄球菌肠毒素B(staphylococcalenterotoxinB,SEB)。方法利用PCR技术从产SEB的金黄色葡萄球菌标准菌株CMCC-26075基因组DNA中克隆SEB全长序列,将其克隆到pGEM-TEasy载体中并进行测序。构建pET-30a-SEB原核表达质粒,转化感受态大肠杆菌BL21(DE3),异丙基硫代β-D半乳糖苷(isopropyl-beta-D-thiogalactopyranoside,IPTG)诱导表达,经镍离子螯合亲和层析纯化后免疫鉴定。结果PCR获得超抗原SEB基因片段,与克隆载体连接后经测序与文献报道的基本一致;成功构建了pET-30a-SEB原核表达质粒且成功诱导表达出相对分子质量(Mr)约31×103的蛋白。结论成功克隆了seb基因序列,并进行了原核表达和鉴定,获得了SEB蛋白,为后续对超抗原SEB的进一步研究奠定了基础。     

Iso-frequency 2-DG contours in the inferior colliculus of the awake monkey

Summary Tone bursts produced bands of selective 2-[¹⁴C]-deoxyglucose labelling in the inferior colliculus (IC) of the awake monkey. Low tone frequencies produced labelling in dorsal regions and high tone frequencies produced labelling in ventral regions. The position of the bands coincided with the position of a single unit with a characteristic frequency, which was the same as the frequency producing the labelling. These findings indicate that the bands of labelling represent iso-frequency contours in IC. The iso-frequency contours extended across most of the nucleus and were oriented from dorsomedially to ventro-laterally at 20–30° from the horizontal and became more vertical anteriorly. The width of the contours was as narrow as 200 m, suggesting that the contours might represent 2 or 3 overlapping cellular laminae.Supported by research grants from the National Health and Medical Research Council of Australia and the Australian Research Grants Scheme