High level expression of recombinated hepatocyte growth factor in human umbilical cord mesenchymal stem cells

AIM: To construct a lentiviral vector encoding human hepatocyte growth factor (hHGF) for transfection,and to observe the expression of hHGF in human umbilical cord mesenchymal stem cells.METHODS: pUC-SRα/hHGF was subcloned into the expression vector pWPI to construct recombinant pWPI-hHGF.hHGF was identified by gene sequence.Recombinant lentivirus was produced by pWPI-hHGF,pAX2 and pMD2G altogether transient transfection into 293T cells using calcium phosphate method.The pWPI-hHGF and the contructed pWPI-GFP were transfected into human umbilical cord mesenchymal stem cells by the Lipofectamin 2000.Through counting by the fluorescent microscope,the efficiency of the transfection was identified.The expressions of hHGF and GFP in human umbilical cord mesenchymal stem cells were also detected.The concentration of hHGF in cell culture medium was determined by enzyme linked immunosorbent assay (ELISA).RESULTS: DNA sequence showed that hHGF cDNA was correctly inserted into pWPI vector.The positive rate of hHGF transfecting 293T cells was 100 %.Bright green fluorescence in the transfected cells was observed under the fluorescent microscope after 24 h transfection with lentiviral plasmid pWPI-hHGF-GFP,and the transfection rate reached 80%.The difference was distinct between the pWPI-hHGF group and control group in the secretive level of hHGF by Western blotting and the ELISA (P<0.01).CONCLUSION: The recombinant pWPI-hHGF plasmid was successfully constructed and efficient,stable and ectopic expression of hHGF was accomplished in human umbilical cord mesenchymal stem cells.     

Inhibition of calcineurin is involved in cardioprotection induced by ischemic postconditioning in rats

AIM: To demonstrate the mechanisms underlying cardioprotection induced by ischemic postconditioning (I-postC) via studying the alteration of calreticulin (CRT)/calcineurin (CaN) signaling pathway in rat heart subjected to ischemia/reperfusion (I/R).METHODS: The model of myocardial I/R injury in vivo was made by occluding the left anterior descending artery for 45 min followed by 24 h of reperfusion in Wistar rats.Hemodynamics and activity of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) in plasma were measured.Myocardial infarct size was measured by 2,3,5- triphenyltetrazolium chloride (TTC) staining and cardiomyocyte apoptosis was detected using in situ TDT-mediated dUTP nick end labeling (TUNEL).The activity of CaN,the expressions of CaN and CRT in myocardium were detected by enzyme reaction phosphorus measurement and Western blotting analysis,respectively.RESULTS: Cyclosporin A,the inhibitor of CaN,limited significantly myocardial infarct size and cardiomyocyte apoptosis induced by I/R,but had no significant effect on cardiac function.I-postC ameliorated significantly the cardiac dysfunction induced by I/R.Compared with those in I/R group,the myocardial infarct size,the LDH and CK-MB activities in plasma and the cardiomyocyte apoptotic index were significantly reduced in I-postC group.In addition,I/R-induced upregulation of CaN activity,CaN and CRT expression were relieved by I-postC.No significant difference was found between I-postC and ischemic preconditioning groups.I-postC had stronger protective effect on the reperfused heart compared with cyclosporin A.CONCLUSION: The findings indicate that I-postC protects myocardium against I/R injury,at least in part,via inhibiting the CRT/CaN signaling pathway.     

甲氰菊酯在豌豆中残留动态及安全使用技术

采用气相色谱法及田间试验方法,研究了甲氰菊酯在豌豆中的残留消解动态,并对其安全使用技术进行试验。结果表明,甲氰菊酯在豌豆中原始沉积量因不同施药处理有较大的差别,施用量越大原始沉积量也越高;残留消解动态均符合一级动力学方程,不同施用量的降解速率基本相似,降解系数（︱k︱）=0.167 8±0.001 3,半衰期（DT₅₀）为4.1～4.2 d,降解99 %所需要的时间（T₉₉）为27.1～27.7 d。按常规方法施用甲氰菊酯有效成分5.00 g·（667 m²）^-1,施药1次或间隔7 d连续施药2次,末次施药后第14天的残留量均＜0.5 mg·kg^-1,第24天的残留量均＜0.1 mg·kg^-1,产品质量安全水平分别符合我国叶菜类蔬菜MRL和日本豆类蔬菜MRL的要求。     

外源ABA对NaCl胁迫下黄瓜幼苗不同离子含量的影响

研究了外源ABA对NaCl胁迫下黄瓜幼苗不同器官中Na+、Cl-、K+和Ca2+含量的影响。结果表明,NaCl胁迫下,黄瓜幼苗体内Na+、Cl-含量升高;叶片中K+、Ca2+含量下降,Na+/K+、Na+/Ca2+增大,SK、Na、SCa、Na降低。外源ABA减少了叶片、根中Na+、Cl-的积累,抑制了叶片K+、Ca2+含量的降低,提高了K+、Ca2+的吸收和运输选择性。外源ABA可通过调控盐胁迫下植株体内的离子平衡,提高黄瓜幼苗的耐盐性。     

硅藻土基质配方对盆栽观赏辣椒生长的影响

研究了五彩椒在硅藻土、草炭、蛭石、珍珠岩4种组分按不同比例配成的5种混合基质下的生长差异,试验采用了一种自制的自动连续精确控水盆栽装置,并为所有处理设置了相同的供水吸力,用于分析5种不同基质的吸水能力.结果表明:硅藻土是一种优良的基质添加成分,其吸水、保水性能均优于蛭石和珍珠岩;试验同时得出:50%草炭+25%蛭石+25%硅藻土、40%草炭+20%蛭石+20%珍珠岩+20%硅藻土2种基质配方比其它基质配方更理想,相对于其它3种基质配方,这2种配方基质理化性质更好,吸水、保水、保肥能力都强于其它基质量配方,栽培下的五彩椒生理指标和形态指标更优,值得进一步研究与推广.     

Characterization of porcine circovirus type 2 (PCV2) infection in swine lymphocytes using mitogen-stimulated peripheral blood lymphocytes from healthy PCV2-carrier pigs

Information regarding the susceptibility of swine lymphocytes to PCV2 is rather limited. To further explore and characterize the PCV2 infection in swine lymphocytes, an in vitro model using concanavalin A (Con A)-stimulated peripheral blood lymphocytes (PBLs) obtained from clinically healthy PCV2-carrier pigs was introduced. It was found that the PCV2 antigen-containing rate was below 2% in PBLs from healthy PCV2-free pigs following treated simultaneously with Con A and PCV2. However, significantly higher PCV2 antigen- and nucleic acid-containing rates could be seen in Con A-stimulated PBLs from clinically healthy PCV2-carrier pigs. Prior to Con A treatment, both of the PCV2 antigen- and nucleic acid-containing rates in PBLs from healthy PCV2-carrier pigs were less than 1%; however, they reached 22.1+/-5.7% by flow cytometry and 27.1+/-6.5% by in situ hybridization, respectively, at 4-day post-incubation with Con A. Phenotyping of PCV2 antigen-containing cells revealed that PCV2-positive cells could be detected in both T and B lymphocyte populations within which IgM-positive B lymphocytes appeared to have a relatively higher positive rate. The Con A-stimulated PBLs also displayed a significantly higher viral load by the measurement of either PCV2 DNA copy number or viral titer when compared with the non-treated PBLs from healthy PCV2-carrier pigs. The results indicate that PBLs, especially IgM-bearing B lymphocytes, are indeed susceptible to PCV2 infection and PCV2 is capable of replicating in dividing lymphocytes. This activation-induced replication may explain in part the pathogenesis of lymphoid depletion in PMWS-affected pigs.     

中国粮食安全问题:警钟已经敲响

<正>2007年5月,我国猪肉价格突然猛涨。在猪肉的带动下,牛羊肉、禽肉、蛋、奶、鱼等动物性食品价格全面上涨。与此同时,粮、油、蔬菜、水果等农产品也纷纷涨价。在市场经济体制下,商品价格有所波动本不足为怪。然而,此次农场品价格上涨的确非     

Changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are pathogens, which can significantly affect the swine industry worldwide. Field surveys suggest that simultaneous PRRSV and PCV2 infection is common in pigs. The objective of this study was to measure the changes in peripheral blood leukocyte subpopulations in piglets co-infected experimentally with PRRSV and PCV2, in order to analyze the synergistic influence of co-infection on the immune system. Changes in peripheral blood leukocyte subpopulations were systematically measured by flow cytometry (FCM). The levels of antibodies to PRRSV and PCV2 were detected by indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) and the indirect fluorescent antibody test (IFA), respectively. Serum viral loads were measured using real-time PCR. The results showed that piglets co-infected with PRRSV and PCV2 exhibited slower generation and lower levels of antibodies to PRRSV and PCV2, and increased amounts and a prolonged presence of both PRRSV and PCV2 in serum, in comparison to the piglets infected with either virus alone. The major finding in our study was that the total and differential leukocyte counts, including white blood cells (WBCs), monocytes, granulocytes and lymphocytes (T, B and NK cells, as well as T-cell subpopulations), dramatically decreased early during co-infection with PRRSV and PCV2 for about two weeks, in contrast with animals singly infected with either PRRSV or PCV2. These results suggest that PRRSV and PCV2 co-infection results in a synergistic decrease in immune cells in the peripheral blood of piglets. These data contribute to the understanding of the immunosuppressive effects resulting from PRRSV and PCV2 co-infection in pigs.     

猪链球菌种及其主要致病血清型多重PCR检测方法的建立

根据猪链球菌谷氨酸脱氢酶基因和血清型1型、2型、1/2型、7型、9型和14型的荚膜多糖编码基因核酸序列,分别设计猪链球菌种和血清型特异性引物,建立并优化多重PCR检测方法,检测分析种属背景明确的73株菌株（其中猪链球菌49株、其他对照菌株24株）及临床分离样本94株（包括四川资阳临床分离样本45株）。其中73株种属背景明确菌株多重PCR种检测结果符合率为87.5%,6种主要致病血清型检出率可达100%。24株对照菌株在种和血清型检测均为阴性。对45株四川猪链球菌病暴发现场分离菌株进行检测,其中41株为猪链球菌2型。上述结果提示建立的多重PCR方法对猪链球菌种及主要致病血清型的检测具有较好的特异性和敏感性,可用于猪链球菌病的快速诊断和流行病学调查。     

应用4个繁殖性状候选基因对太湖猪群体遗传结构分析

采用PCR-RFLP技术分析雌激素受体(ESR)基因、促卵泡素β亚基(FSHβ)基因、催乳素受体(PRLR)基因和视黄醇结合蛋白4(RBP4)基因在太湖猪不同类群中的分布情况。结果表明:ESR基因在小梅山猪、二花脸猪、枫泾猪中B基因频率较高,分别为0.629,0.762,0.649;FSHβ基因在小梅山猪、二花脸猪、枫泾猪中AA基因型频率较高,分别为0.875,0.881,0.833;PRLR基因在小梅山猪、二花脸猪、枫泾猪中AA基因型频率分别为0.631,0.571,0.281;RBP4基因在小梅山猪、二花脸猪、枫泾猪猪群中AA基因型频率较高,分别为0.593,1.000,0.900。对不同基因的特性分析表明:ESR、FSHβ、PRLR和RBP4在3个类群中的多态信息含量和杂合度均不高,大多为中低等多态位点。