玉米秸秆残茬覆盖对大豆田土壤水温效应的影响

研究了传统耕作、玉米原垄留茬耕作、玉米原垄留茬全覆盖、玉米留茬条带覆盖、浅松覆盖和深松覆盖6种耕作方式下土壤含水率、棵间蒸发量和地温的变化。结果表明,条带覆盖对大豆不同生育期土壤剖面含水率影响最大;其他耕作方式对棵间蒸发的抑制作用均小于传统耕作;日平均地温在苗期变化明显;保护性耕作对日平均地温影响较大,传统耕作最高,留茬全覆盖最低。     

The Fluorescent Characterization and Analysis of Two Brucella Attenuated Vaccine Infecting Mouse Macrophagocyte

The experiment was conducted to discuss the difference of binding time of green fluorescent protein B.melitensis M5 (GFP-M5) and B.abortus S19 (GFP-S19) infecting the mouse macrophagocyte (RAW264.7),lysosome,endoplasmic reticulum and golgi body in the initial stage and compare the binding rate of GFP-M5,GFP-S19 with organelle in different timeline,respectively,by confocal laser scanning microscope (CLSM) and flow cytometry.The result showed that GFP-M5 and GFP-S19 were successfully constructed.The intracellular survival ability of Brucella M5,Brucella S19,GFP-M5 and GFP-S19 were not obvisouly affected after infecting RAW264.7.GFP-M5 and GFP-S19 could enter the macrophagocyte in 30 mins,and in 2 h the Brucella could reach lysosome,endoplasmic reticulum and golgi body.In addition,the binding time for two attenuated vaccine did not show differences in 1,2,3 and 4 h.The content of GFP⁺ cell produced by RAW264.7 infected by GFP-M5 and GFP-S19 did not show significant differences (P>0.05).Therefore,the two strains did not have significant differences in the invasion ability in the initial stage of infecting host cell.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.     

Analysis on Differential Expression Genes in Porcine Aortic Vascular Endothelial Cells Induced by Apolipoprotein C Ⅲ

The aim of this study was to investigate the differential expression genes induced by ApoCⅢ,and study the function of ApoCⅢ.Porcine aortic vascular endothelial cells were successfully isolated using enzyme digestion,and then screened the differential expression genes induced by ApoCⅢ using the Solexa high-throughput sequencing technology.The results showed 647 differential expression genes,including 390 up-regulated genes and 257 down-regulated genes.The qRT-PCR results verified that the gene expression results from Solexa sequencing data were reliable.GO and Pathway analysis showed that the function of differential expression genes were related to immune response,cell apoptosis and death.These findings suggested that ApoCⅢ affected the physiological function of porcine aortic endothelial cells by the molecular pathways of inflammation,cell adhesion and apoptosis,which provided a theoretical basis for further understanding the molecular mechanisms of atherosclerosis caused by ApoCⅢ.     

Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43

The aim of this study was to determine the effect of vitamin E on Cx43,the mechanism and function of vitamin E on bovine granulosa cells apoptosis and proliferation.In this study,granulosa cells were isolated from bovine ovary and cultivated in vitro by adding different concentration of vitamin E (0,25,50,100,200 and 500 μmol/L) for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53 and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100 μmol/L vitamin E could significantly inhibit the apoptosis of granulosa cells (P<0.05).Real-time PCR detection results showed that vitamin E significantly changed the mRNA expression levels of BCL2/BAX,P53,Cx43 genes (P<0.05).Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36 h (P<0.05).The results provided a theoretical basis on further analysis for studing the influence mechanism of vitamin E on oocytes development and maturity,and improvement of female animal reproduction by influencing granulosa cells proliferation and apoptosis.     

Development of A Dual Real-time PCR for the Rapid Detection of Listeria monocytogenes

To establish a rapid assay for Listeria monocytogenes(LM) detection,a Real-time PCR method was developed targeting iap gene of LM.The results showed that the test for 15 bacteria strains,only LM was positive,indicated that the method had high specificity.In addition,the sensitivity of Real-time PCR was 6.5 CFU/mL.Stability and reproducibility of the test showed that the coefficient of variation for the same sample repeat the Ct values were less than 2%.Furthermore,a total of 3 positive samples for LM were detected from 139 clinical samples by the method,which was in accordance with the testing result by GB 478930-2010 standard detection protocol.Therefore,the Real-time PCR method provides a novel rapid,sensitive and good repeatability detection method for LM infection.     

Study on Fans Cooling of Dairy Cowshed with Free Stall in Northern Mountainous Region of Hebei in Summer

The present objective of the study was to investigate the effect of fans cooling on dairy cow in northern mountainous region of Hebei in summer.Two cowsheds with the same structure and feeding technics were used to investigate the effect of fans on indoor temperature and humidity, physiological parameters and milking performance of dairy cow.The results showed that the average ambient temperature in cowshed with fans was 1.5℃ lower than that of the control cowshed, and the average wind velocity was 0.49 m/s higher than that of the control cowshed, reaching to 0.66 m/s.Temperature-humidity index (THI) of cowshed with fans was significantly lower than that of the control shed during the period of fans running (P<0.05).Besides, the rectal temperature and respiration rate of dairy cows in cowshed with fans were significantly lower than of the control cowshed (P<0.05), and the milk yield per cow per day was 17.0% higher than that of the control cowshed (P<0.05), however, the milk fat, milk protein and milk sugar were no significant difference from the control (P>0.05).Overall, fans could decrease indoor environment, moderate heat stress and increase performance and health of dairy cows in northern mountainous region of Hebei.     

西瓜嫁接砧木对南方根结线虫的抗性鉴定

以16种西瓜砧木为试材,采用聚类分析和隶属函数分析方法,研究了南方根结线虫(Meloidogyne incognita)对不同砧木的生长指标和抗病指标的影响。结果表明:抗性最强的是"青农2号",隶属函数总值为6.26,抗性最弱的是"青农3号",隶属函数总值为1.48。结论认为"青农2号"为西瓜嫁接的最佳砧木,该试验为合理筛选和利用西瓜砧木提供了理论和技术参考。     

去花对菊芋数量学性状的影响

以白皮菊芋为试材,通过对现蕾期菊芋去除1/4花、1/3花、1/2花、全部去除和不去花等5个处理,测定叶片、小花、分枝和块茎数量等相关指标,探讨了去花对菊芋数量学性状的影响规律,为提高菊芋块茎生物产量提供理论依据。结果表明:去花1/3、1/2和全株去花提高了单株菊芋块茎数量,降低单株块茎质量,1/4去花对菊芋单株块茎数量和单个块茎质量影响不显著;去花处理降低了二级分枝和一级分枝的数量;去花处理增加了单株叶片的数量和单片叶片的质量;随着去花数量的增加,单株小花数量呈现"降-升-降"的变化规律,单个小花质量呈现"降-升"的变化趋势。