Neurexin-1 is required for synapse formation and larvae associative learning in Drosophila

Neurexins are highly polymorphic cell-surface adhesive molecules in neurons. In cultured mammalian cell system, they were found to be involved in synaptogenesis. Here, we report for the first time that Drosophila neurexin is required for synapse formation and associative learning in larvae. Drosophila genome encodes a single functional neurexin (CG7050; Neurexin-1 or Nrx-1), which is a homolog of vertebrate alpha-neurexin. Neurexin-1 is expressed in central nervous system and highly enriched in synaptic regions of the ventral ganglion and brain. Neurexin-1 null mutants are viable and fertile, but have shortened lifespan. The synapse number is decreased in central nervous system in Neurexin-1 null mutants. In addition, Neurexin-1 null mutants exhibit associative learning defect in larvae.     

Assessment of genetic stability of the germplasm lines of medicinal plant <Emphasis Type="Italic">Scutellaria baicalensis</Emphasis> Georgi (Huang-qin) in long-term,in vitro maintained cultures

An approach of combining flow cytometry (FCM) analysis with morphological and chemical profiling was used to assess the genetic stability and bioactive compound diversity in a Scutellaria baicalensis Georgi (Huang-qin) germplasm collection that was clonally maintained in in vitro for a period of over 6 years. Based on the FCM analysis of nuclei samples from young shoots, the nuclear DNA content of S. baicalensis was calculated as 0.84 pg/2C. FCM analysis showed no significant variation in the nuclear DNA contents and ploidy levels in the long-term in vitro maintained germplasm lines. Germplasm lines, acclimatized to ex vitro conditions, exhibited distinctive plant growth and bioactive compound production capacities. The high level of genetic stability observed in in vitro maintained S. baicalensis lines opens up a variety of opportunities such as allowing long-term aseptic preservation and easy distribution of well-characterized germplasm lines of this medicinal plant species. This study represents a novel approach for continuous maintenance, monitoring, and production of medicinal plant tissues with specific chemistry.     

海南岛尖峰岭半落叶季雨林“刀耕火种”生态后果的初步观测

热带地区的降水时间集中,降水强度大,一旦森林破坏地表裸露后,很容易引起生态性危害,山地尤甚。本文通过观测和对比试验,初步探讨了海南岛尖峰岭半落叶季雨林的生态效益及流动农业所造成的生态恶果。毁林耕垦当年即可使地区性小气候恶化、水土流失严重、地力开始减退。     

Serological diagnosis of <Emphasis Type="Italic">Mycoplasma pneumoniae</Emphasis> infection by using the mimic epitopes

Nowadays, there is lack of effective serological detection method for Mycoplasma pneumoniae (M. pneumoniae) infection in clinic. In this study, the mimic epitopes of M. pneumoniae were screened to evaluate the role in the serodiagnosis of M. pneumoniae infection. The M. pneumoniae-positive serum was used as the target for biopanning to phage display random 7-peptide library. The positive phage clones were selected and the DNA were sequenced and analyzed by BLAST. The representative phages were identified using dot immunoblotting and ELISA. The exogenous heptapeptides were synthesized and their reactions with M. pneumonia-positive serum were tested by indirect ELISA. Two heptapeptides, namely heptapeptide 1: TVNFKLY and heptapeptide 2: LPQRLRT, were screened out from the randomly selected 40 phages after the four bio-panning rounds. They had high homologies to some M. pneumoniae antigens. Besides, the representative bacteriophage containing heptapeptide 1 or 2 could react with the M. pneumonia- positive serum. The sensitivities of heptapeptide 1 and heptapeptide 2 for the diagnosis of M. pneumoniae infection were 90.1 and 80.0%, respectively, and the specificities were 94.3 and 97.1%, respectively. Therefore the two heptapeptides were the mimic epitopes of M. pneumoniae and might have potential serological diagnosis value for M. pneumoniae infection.     

Genome shuffling and high-throughput screening of <Emphasis Type="Italic">Brevibacterium flavum</Emphasis> MDV1 for enhanced <Emphasis Type="SmallCaps">l</Emphasis>-valine production

l-valine is an essential branched-amino acid that is widely used in multiple areas such as pharmaceuticals and special dietary products and its use is increasing. As the world market for l-valine grows rapidly, there is an increasing interest to develop an efficient l-valine-producing strain. In this study, a simple, sensitive, efficient, and consistent screening procedure termed 96 well plate-PC-HPLC (96-PH) was developed for the rapid identification of high-yield l-valine strains to replace the traditional l-valine assay. l-valine production by Brevibacterium flavum MDV1 was increased by genome shuffling. The starting strains were obtained using ultraviolet (UV) irradiation and binary ethylenimine treatment followed by preparation of protoplasts, UV irradiation inactivation, multi-cell fusion, and fusion of the inactivated protoplasts to produce positive colonies. After two rounds of genome shuffling and the 96-PH method, six l-valine high-yielding mutants were selected. One genetically stable mutant (MDVR2-21) showed an l-valine yield of 30.1 g/L during shake flask fermentation, 6.8-fold higher than that of MDV1. Under fed-batch conditions in a 30 L automated fermentor, MDVR2-21 accumulated 70.1 g/L of l-valine (0.598 mol l-valine per mole of glucose; 38.9% glucose conversion rate). During large-scale fermentation using a 120 m³ fermentor, this strain produced?>?66.8 g/L l-valine (36.5% glucose conversion rate), reflecting a very productive and stable industrial enrichment fermentation effect. Genome shuffling is an efficient technique to improve production of l-valine by B. flavum MDV1. Screening using 96-PH is very economical, rapid, efficient, and well-suited for high-throughput screening.     

Role of protein kinase A in GABAA receptor dysfunction in CA1 pyramidal cells following chronic benzodiazepine treatment

One-week treatment with the benzodiazepine (BZ) flurazepam (FZP), results in anticonvulsant tolerance, associated with reduced GABAA receptor (GABAR) subunit protein and miniature inhibitory post-synaptic current (mIPSC) amplitude in CA1 neurons of rat hippocampus. Because protein kinase A (PKA) has been shown to modulate GABAR function in CA1 pyramidal cells, the present study assessed whether GABAR dysfunction is associated with changes in PKA activity. Two days after 1-week FZP treatment, there were significant decreases in basal (- 30%) and total (- 25%) PKA activity, and a 40% reduction in PKA RIIbeta protein in the insoluble fraction of CA1 hippocampus. The soluble component of CA1 showed a significant increase in basal (100%) but not total PKA activity. Whole-cell recording in vitro showed a 50% reduction in mIPSC amplitude in CA1 pyramidal cells, with altered sensitivity to PKA modulators. Neurons from FZP-treated rats responded to 8-bromo-cAMP with a significant increase (31%) in mIPSC amplitude. Likewise, vasoactive intestinal polypeptide (VIP), an endogenous PKA activator, caused a significant 36% increase in mIPSC amplitude in FZP-treated cells. Neither agent had a significant effect on mIPSC amplitude in control cells. This study supports a role for PKA in GABAR dysfunction after chronic FZP treatment.     

人红细胞生成素单克隆抗体的制备、鉴定及应用研究

用rhEPo作为抗原,免疫BALB／c小鼠,取其脾细胞与x63Ag8．653小鼠骨髓瘤细胞融合,再碱性PAGE方法进一步分离并纯化的rhEpo,包被Pvc板,对杂交瘤用ELlSA方法进行筛选,获得两株稳定分泌抗hEPO单抗的杂交瘤细胞株。经鉴定分别属于IgG1、IgG2b,轻链均为k链,Kd分别为5．53×10^-10mol／L和1．34×1O^-10mol／L．用western blot方法证明两者对hEPO具有高度韵专一性．能特异地识别rhEPO和尿源hEPO。所制备单抗可作为亲和层析的配体,用于再生障碍性贫血病人尿中EPO及哺乳类工程细胞所表达的hEPO的分离、纯化,并可用于hEPO的定量检测．     

华癸中慢生根瘤菌7653R 3个脂质转运蛋白基因的克隆、突变及共生表型北大核心CSCD

【目的】脂类转移家族蛋白基因编码一类参与脂类转运及代谢的蛋白。本研究旨在构建华癸中慢生根瘤菌3个脂质转运家族蛋白基因的突变株,检测及分析突变体与紫云英共生条件下的表型及功能。【方法】利用生物信息学分析与预测转脂蛋白的结构特征及功能,采用荧光定量技术检测目标基因在自生和共生条件下的表达特性,通过插入突变技术构建目标基因突变株,并进行植物盆栽实验考察其共生表型。【结果】MCHK-5577、MCHK-2172和MCHK-2779基因编码蛋白属于START/RHO alpha_C/PITP/Bet_v1/Cox G/Cal C(SRPBCC)超家族,包含脂类转移结构域,参与脂类转运或代谢,与百脉根等中慢生根瘤菌相应基因的序列相似性达95%以上。这3个基因在共生条件下的表达水平都增高。分别构建了MCHK-5577、MCHK-2172和MCHK-2779基因突变菌株,与野生型菌株7653R相比,接种突变株MCHK-2172mut、MCHK-2779mut和MCHK-5577mut后的植株地上部分生物量和根瘤固氮酶活性显著降低。【结论】华癸中慢生根瘤菌脂质转移家族蛋白基因在共生互作过程中发挥重要作用,突变后明显影响共生固氮表型。本文的实验结果为深入研究脂类转移蛋白在共生固氮作用中的功能机制奠定了基础。