Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Effects of Momordica grosvenori Saponins on Type 2 Diabetes Rats Induced by High-sucrose/High-fat Diet and Streptozotocin

The purpose of the experiment was to study the effect of Momordica grosvenori saponins on improving the effect of high-sucrose/high-fat diet combined with streptozotocin induced type 2 diabetes in rats.The type 2 diabetes rat model was established with a high-sucrose and high-fat diet combined with streptozotocin.Successfully modeled rats were given the total saponins of Momordica grosvenori (100,200,400 mg/kg) or metformin (268 mg/kg) by gavage,once a day for 4 weeks.Recording the "three more and one less"index (24 h internal urine output,water intake and feed intake,body weight);Detecting of biochemical indicators,analysis of fasting blood glucose (FBG),serum total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),insulin (INS),catalase (CAT),superoxide dismutase (SOD),malondialdehyde (MDA),alanine aminotransferase (AST),aspartate aminotransferase (ALT),blood urea nitrogen (BUN) and creatinine (Cr) levels.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the pancreas.Compared with the model group,the feed intake,water intake,urine output,blood indicators FBG,INS,MDA,TC,TG,BUN and Cr levels,AST and ALT activities were significantly or extremely significantly decreased in the total saponins of Momordica grosvenori each dose group and metformin group (P<0.05;P<0.01),while CAT,SOD activities and HDL-C level were significantly or extremely significantly increased (P<0.05;P<0.01).In pancreatic tissue,the number of pancreatic cells was increased significantly,and the pathological damage of the pancreas was significantly reduced. Momordica grosvenori saponins and metformin had obvious effects on improving glucose metabolism disorders,and at the same time could significantly reduce insulin resistance,enhance the body’s ability to resist oxidative stress and improve liver and kidney functions.     

近年来我国野生动植物进出口贸易中的问题分析及对策

通过对2000年、2002年野生动植物进出口贸易额的比对分析，对野生动植物进出口贸易中有关贸易额、贸易价格、中医药、海关记录数据等主要问题进行了具体分析，并对加强野生动植物的进出口管理提出几点建议：规范海关记录、监管标准；加强大宗贸易的审查和管理；推广限额、限价管理；进行全国野生动植物贸易调查；争取某些产品的特惠税制度等。     

精子受精抗原-1(FA-1) mRNA在绵羊睾丸和附睾中的表达

本研究通过RT-PCR检测了该基因在绵羊睾丸和附睾中的表达情况。首先,提取绵羊睾丸组织、附睾头、体、尾部组织总RNA,以此为模板,反转录合成cDNA,自行设计引物,PCR扩增出462 bp的目的DNA。然后,将目的片断克隆入T载体,通过菌液PCR和重组质粒酶切,鉴定重组质粒中的目的DNA。再经序列分析鉴定目的片断。同时,以β-actin为内参照物,进行RT-PCR半定量分析,比较精子受精抗原(FA-1)在睾丸、附睾头、体、尾组织中的表达量。结果表明,FA-1在绵羊睾丸和附睾中均表达。     

Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/PLGC group. However, the new cortical bone in the TCP/PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.     

EHEG O157△ler/stx(pBR322∷stx 1/2B∷eae)的构建与生物学特性分析

为了获得安全性好,对出血性大肠杆菌具有良好免疫保护作用的疫苗株,本实验以构建的O157△ler/stx基因缺失突变株为载体,将决定特异性粘附的eae全基因插入到质粒pBR322∷stx1/2B中,构建了基因工程菌O157△ler/stx(pBR322∷stx1/2B∷eae),对该基因工程菌进行生物学特性分析,实验结果表明,该菌正常培养可表达eae;具有粘附HEp-2细胞的特性;该菌培养上清对Vero细胞没有毒性作用;给小鼠口服接种该工程菌后可在肠道中定居至少10 d;14日龄小鼠口服活菌量为5×109CFU/只,小鼠生长正常,没有出现任何临床症状;该菌通过口服免疫小鼠可以诱导肠道粘膜免疫应答和体液免疫应答.这些结果表明,该工程菌安全性好,免疫原性强,是有价值的预防EHEC O157感染的基因工程疫苗候选株.     

Evaluation of turnip-rooted parsley (Petroselinum crispum ssp. tuberosum) for root and foliage production under a warm,Mediterranean climate

Two experiments were undertaken in southern Greece during 2000–2002 in which turnip-rooted parsley was tested for its suitability for cultivation in comparison with locally grown curly-leafed and plain-leafed forms. Plants were harvested when the foliage was acceptable for marketing, rather than on completion of the vegetative cycle, hence root size was relatively small. In year 1, the foliage development of plain-leafed parsley was significantly higher than that of the other two forms, irrespective of sowing date. In year 2, both plain-leafed and curly-leafed parsley produced more foliage than turnip-rooted parsley in the early sowing (November), but not in subsequent sowings (December and February). During both years, root development was higher in turnip-rooted parsley than in curly-leafed parsley, although in the November and February sowings of year 1 the root weight of both plain-leafed and turnip-rooted forms were similar. In all three types of parsley, foliage yield fell off significantly in the spring sowing (February), and in year 1 there was a considerable decrease in root production as well. In most cultivars, foliage weight was higher when plants were grown from transplanted seedlings rather than by direct sowing, whereas the effect of sowing method on root weight was sowing date dependent. Cultivar differences in the foliage weight of direct sown and transplanted plants were reflected by differences in plant height and, in some cases, by leaf number. It is concluded that although overall turnip-rooted parsley produced rather less foliage than that plain-leafed parsley, this was compensated for by the satisfactory production of roots and therefore turnip-rooted cultivars may constitute a suitable crop for introduction to the Mediterranean region.     

西瓜柱头ATP酶活性及分泌活动研究

用光学显微镜和电子显微镜研究了西瓜柱头结构、ATP酶活性和分泌活动。结果表明,柱头属于湿柱头类型,最外层是乳突细胞,内层为薄壁组织。授粉后10min,乳突细胞质膜和胞间连丝上ATP酶活性强;30min后,质膜上ATP酶活性极弱,但液泡膜上有很强的ATP酶活性;授粉1h后,ATP酶活性很难检测到。分泌活动始于开花前1d;开花后,授粉的柱头分泌活动急剧增强,授粉10min后分泌物最多,30min后不再产生新分泌物;1h后,细胞解体;24h后分泌物变干燥;未授粉的柱头分泌活动没有激增过程。     

氧氟沙星在肉鸡体内残留的研究

建立了氧氟沙星在肉鸡组织内残留的高效液相色谱分析法。采用荧光检测器，激发波长278nm，发射波长445nm。色谱柱为Hypersil BDS—C18，柱温40℃；以磷酸（三乙胺调pH至2．5）-乙腈（86：14）为流动相，流速1．0mL／min；进样量为20μL。氧氟沙星在肌肉、肝脏和肾脏中的平均回收率分别为96．3％、87．5％和76．4％，检测限为10ng／g，在10～1000ng／mL浓度范围内，氧氟沙星的峰面积与其浓度呈良好的线性相关，达到了氧氟沙星残留分析的要求。利用建立的方法对氧氟沙星在肉鸡体内的残留消除规律进行了研究，以1L水添加500mg的剂量，对120只肉鸡连续给药5d，对不同停药时间所取肌肉、肝脏和肾脏样品中的药物残留进行分析。结果表明，氧氟沙星在肉仔鸡组织中的残留消除速度较为缓慢，至休药23d时，仍可在肝脏和肾脏分别检出39和35μg/kg的氧氟沙星残留。在本实验的条件下，建议氧氟沙星临床休药期以25d为宜。     

Distribution and identification of immunocytes in humanized SCID mouse model

AIM: Humanized-NOD/SCID(hu-NOD/SCID) mouse model was established and the level of immune reconstitution was assessed in this model. METHODS: Mononuclear cells (MNC) and CD34⁺ cells were isolated or sorted from cord blood(CB). Human CD45, CD19, CD3 markers on cells from NOD/SCID murine peripheral blood(PB), bone marrow(BM), thymus were detected by FCM from 4 to 10 weeks after hematopoietic stem cell transplantation. After 10 weeks, the gene expressions of the human β₂M and RAG2 were detected by RT-PCR in PB or bone marrow of mice model. RESULTS: Human CD45, CD19, CD3 cells populations in PB and BM were found by flow cytometry in mice model transplanted with CD34+ cells or CB MNC from 4 to 10 weeks. The highest positivity of human lymphocytes was at 8 week after transplantation. The levels of human cell engraftment in mice transplanted with CD34⁺ cells were higher than those in mice transplanted with CB MNC. The mRNA of human β₂M and RAG2 were found by RT-PCR in BM.CONCLUSION: The higher level of human lymphocyte engraftment is established in NOD/SCID mouse model transplanted with CD34⁺ compared with CB MNC. The maturation of T lymphocytes could be happened in bone marrow of mice model.