The p75 neurotrophin receptor is a central regulator of glioma invasion

The invasive nature of cancers in general, and malignant gliomas in particular, is a major clinical problem rendering tumors incurable by conventional therapies. Using a novel invasive glioma mouse model established by serial in vivo selection, we identified the p75 neurotrophin receptor (p75^NTR) as a critical regulator of glioma invasion. Through a series of functional, biochemical, and clinical studies, we found that p75^NTR dramatically enhanced migration and invasion of genetically distinct glioma and frequently exhibited robust expression in highly invasive glioblastoma patient specimens. Moreover, we found that p75^NTR-mediated invasion was neurotrophin dependent, resulting in the activation of downstream pathways and producing striking cytoskeletal changes of the invading cells. These results provide the first evidence for p75^NTR as a major contributor to the highly invasive nature of malignant gliomas and identify a novel therapeutic target.     

Relationships between adenylyl cyclase activity,glucose metabolism,and both intestinal insulin-like peptide and adipokinetic hormone in mealworm larval fat body

Adenylyl cyclase activity in mealworm larval fat body and its dependence on external Ca²⁺ concentrations and glucose catabolism pathways in vitro were analyzed for their response to intestinal insulin-like peptide (ILP) and synthetic locust adipokinetic hormone I (AKH-I). Dose-response curve for cAMP accumulation was obtained with increasing doses of ILP. The effects of ILP were ten-fold lower than those of AKH-I. Forskolin and IBMX provoked very high levels of cAMP accumulation but IBMX did not potentiate the effects of ILP on cAMP accumulation. Accumulation of cAMP after a treatment with ILP involves a Ca²⁺ influx via the opening of L-type voltage gated channels. The cyclic AMP analogue chlorophenyl thio-cAMP as well as AKH-I induced a decrease in the utilization of the pentose cycle while the effects of ILP were totally opposite. Finally, the antagonism between ILP and AKH-I on glucose metabolism results from a participation of different transduction pathways (Ca²⁺-dependent and Ca²⁺-independent, respectively), probably located in the same target cells, thus suggesting the involvement of mechanisms similar to those existing in mammals. © 1996 Wiley-Liss, Inc.     

Box-Behnken响应面法优化司替霉素B产生菌发酵条件

【背景】粗糙链霉菌(Streptomyces scabrisporus) HBERC-53204是本中心自主分离的一株链霉菌,经鉴定,其产生一种活性化合物司替霉素B (steffimycin B,SMB),对多种动植物重要病原菌具有良好生物活性。【目的】提高SMB发酵水平,拓宽放线菌活性天然产物在农牧业领域的研究及应用。【方法】以本实验室筛选出的一株产SMB的粗糙链霉菌HBERC-53204为研究对象,运用单因素试验筛选培养基的主效碳源、氮源、无机盐及各营养成分最适浓度,并基于单因素试验结果,通过Plackett-Burman(PB)试验设计筛选出显著影响因素,再结合最陡爬坡试验、Box-Behnken (BB)响应面法拟合显著因子与产量的非线性方程求解,进一步优化菌株产SMB的最佳发酵培养基配方。【结果】优化后最佳培养基配方为：葡萄糖36.22 g/L,蛋白胨8.00 g/L,酵母粉8.51 g/L,酸水解酪蛋白1.50 g/L,MgSO₄ 0.68 g/L,KNO₃ 1.00 g/L。经摇瓶验证,优化后SMB效价达到477.26 mg/L...     

Radioimmunoassay of urinary 21-deoxytetrahydroaldosterone in primary aldosteronism and 21-hydroxylase deficiency

A radioimmunoassay of 21-deoxytetrahydroaldosterone was developed. Normal daily excretion of the unconjugated metabolite was 1.2 +/- 1.3 micrograms and of the glucuronized metabolite, 11.9 +/- 7 micrograms. The tetrahydroaldosterone/21-deoxytetrahydroaldosterone ratio varied more in patients with primary aldosteronism than in control subjects. Thus, measurements of the urinary excretion of the tetrahydroaldosterone or 21-deoxytetrahydroaldosterone alone did not provide an accurate expression for aldosterone production. Their sum correlated well with the clinical condition, i.e. clear-cut elevation in patients with primary aldosteronism. The diminished tetrahydroaldosterone/21-deoxytetrahydroaldosterone ratio found in patients with 21-hydroxylase deficiency may be attributed to increased bacterial conversion of tetrahydroaldosterone to 21-deoxytetrahydroaldosterone but could also stem from a deficiency implicating zona glomerulosa (aldosterone biosynthesis) regardless of the stage and clinical presentation of the disease.     

混合盐碱胁迫下接种丛枝菌根真菌和根瘤菌对紫花苜蓿生长的影响

随着气候变化,全球土壤盐碱化严重威胁着农牧业生产和生态环境的建设。土壤盐化与碱化常常伴随发生,目前多数研究侧重于盐胁迫,忽略了碱胁迫的存在。实际上,碱性盐除了盐度外还伴随着高p H。由此,试图探讨混合盐碱胁迫下接种丛枝菌根真菌(AMF)和根瘤菌对紫花苜蓿生长的影响及其作用机理。根据中国盐碱地的特点,将4种盐(Na Cl、Na_2SO_4、Na_2CO_3、Na HCO_3)按一定比例混合模拟出16种混合盐碱(盐浓度25—150 mmol/L,p H 7.67—10.52)胁迫浓度,按照L_(16)(16~1×2~2)混合正交设计,采用盆栽法研究了混合盐碱胁迫下接种AMF摩西管柄囊霉(Funneliformis mosseae)和根瘤菌(Sinorhizobium meliloti)对紫花苜蓿生长及生理的影响。结果表明:对紫花苜蓿株高的影响程度依次是AMF+根瘤菌pHAMF盐碱类型根瘤菌;对地上生物量、地下生物量、茎高净增长量、丙二醛(MDA)及脯氨酸影响程度依次是p HAMF+根瘤菌盐碱类型AMF根瘤菌;对超氧化物歧化酶(SOD)的影响程度依次是p HAMF+根瘤菌盐碱类型AMF根瘤菌;对过氧化物酶(POD)影响程度依次是p HAMF+根瘤菌根瘤菌AMF盐碱类型;对蛋白质的影响程度依次是p H盐碱类型根瘤菌AMF+根瘤菌=AMF。株高、地下生物量、POD均在双接种的处理下出现最大值。本研究证明:盐碱混合胁迫会严重抑制植物的生长发育;而接种AMF和/或根瘤菌均可有效缓解盐碱胁迫对植物造成的伤害,提高植物对盐碱的耐受能力;且同时接种AMF及根瘤菌效果最好,其次是单独接种AMF,单独接种根瘤菌作用相对最小。     

植物对盐碱胁迫的响应机制研究进展

盐碱胁迫是制约植物生长发育的主要非生物胁迫之一,也是制约农作物生产和生态环境建设的严峻问题。研究作物的耐盐碱机理,对开发和有效利用盐碱地有重要的现实意义。许多研究将盐碱胁迫笼统称为盐胁迫,实际上这是两种不同的非生物胁迫,且碱胁迫对植物的伤害要大于盐胁迫。总结性阐述了盐碱胁迫对植物的危害。从生物量、光合作用、离子平衡和膜透性等方面分析了植物对盐碱胁迫的响应机制,并结合最新研究从多角度综述了植物的抗盐碱机理,包括合成渗透调节物质、提高抗氧化酶活性、对离子的选择性吸收及p H平衡和诱导抗盐碱相关基因表达。提出了抗盐碱性的途径,即外源物质的加入、与真菌的协同效应、利用生物技术手段、培育耐盐碱品种和抗性锻炼。最后针对植物适应盐碱逆境方面的研究进行了展望,提出了当前研究需要解决的问题和突破口,旨在为提高植物耐盐碱能力、增加作物产量提供一定的理论依据。     

A simple nanobody-based competitive ELISA to detect antibodies against African swine fever virus

African swine fever virus (ASFV) infection is a big threat to the global pig industry. Because there is no effective vaccine, rapid, low-cost, and simple diagnosis methods are necessary to detect the ASFV infection in pig herds. Nanobodies, with advantages of small molecular weight and easy genetic engineering, have been universally used as reagents for developing diagnostic kits. In this study, the recombinant ASFV-p30 was expressed and served as an antigen to immunize the Bactrian camel. Then, seven nanobodies against ASFV-p30 were screened using phage display technique. Subsequently, the seven nanobodies fused horseradish peroxidase (nanobody-HRP) were secretory expressed and one fusion protein ASFV-p30-Nb75-HRP was selected with the highest sensitivity in blocking ELISA. Using the ASFV-p30-Nb75-HRP fusion protein as a probe, a competitive ELISA (cELISA) was developed for detecting anti-ASFV antibodies in pig sera. The cut-off value of cELISA was determined to be 22.7% by testing 360 negative pig sera. The detection limit of the cELISA for positive pig sera was 1:320, and there was no cross-reaction with anti-other swine virus antibodies. The comparative assay showed that the agreement of the cELISA with a commercial ELISA kit was 100%. More importantly, the developed cELISA showed low cost and easy production as a commercial kit candidate. Collectively, a simple nanobody-based cELISA for detecting antibodies against ASFV is developed and it provides a new method for monitoring ASFV infection in the pig herds.