Cytokine profiles in peripheral blood mononuclear cells from patients with cystic echinococcosis

IntroductionCystic echinococcosis (CE) is a chronic zoonotic disease caused by the larval stage of Echinococcus granulosus (E. granulosus), which affects domestic and wild carnivores as the definitive host and ungulates as intermediate hosts. In intermediate hosts, both Th1 and Th2 cells are involved in the immune responses to an echinoccocal infection. This study aimed to investigate production of IL-4, IL-10, and IFN-γ cytokines in peripheral blood mononuclear cells (PBMCs) of CE patients before and after surgical treatment.MethodsTo evaluate cytokine production in response to E. granulosus antigens, we investigated IL-4, IL-10, and IFN-γ production in PBMCs of 20 CE patients in response to hydatid cyst fluid antigen (HCF-Ag) before and after surgical treatment using ELISA.ResultsThe mean IL-4 production from HCF-Ag stimulated PBMCs was significantly decreased (p < 0.05), while IFN-γ was significantly increased in HCF-Ag stimulated PBMCs in patients after surgery (p = 0.005).Furthermore, our results showed that there is no significant difference between IL-10 production in patients before and after treatment (p = 0.562).ConclusionsOur data Indicated production of IL-4 in cultured PBMCs of CE patients stimulated with HCF-Ag was decreased significantly. While, production of IFN-γ was increased significantly in responses to HCF Ag after surgery. We concluded that the evaluation of IL-4 and IFN-γ in HCF-Ag stimulated PBMCs of CE patients should be considered as a useful marker in the follow up of patients with cystic echinococcosis.     

低温对2种紫草科植物△6 脂肪酸脱氢酶基因表达的影响

以紫草科植物新疆软紫草(Arnebia euchroma)和假狼紫草(Nonea capsica(willd.)G.Don)为材料,翻译延伸因子EF1a为内参基因,利用半定量逆转录聚合酶链式反应(semi-RT-PCR)技术,对2种紫草植物中γ-亚麻酸合成关键酶基因△6-脂肪酸脱氢酶基因的表达进行研究。结果表明,△6-脂肪酸脱氢酶基因在紫草植物的叶、茎、根中均表达,在低温诱导不同时间后紫草植物叶片中△6-脂肪酸脱氢酶基因的表达有所增强。气相色谱分析显示,其产物γ-亚麻酸在低温下积累,推测紫草科植物叶片中△6-脂肪酸脱氢酶基因表达量增加可能与适应低温胁迫的逆境有关。     

东北虎γ-干扰素基因的克隆、生物信息学分析及其在毕赤酵母中的表达

本试验旨在通过克隆东北虎γ-干扰素(IFN-γ)基因,研究其分子特征并预测蛋白生物学功能,为后续研究干扰素抗病毒活性做前期准备。通过RT-PCR从ConA诱导过的东北虎血淋巴细胞中扩增东北虎IFN-γ基因并测序,应用生物信息学方法进行序列分析。结果表明:东北虎IFN-γ编码区由504个核苷酸组成,共编码167个氨基酸,蛋白相对分子质量为19.59 ku,等电点为9.03,所编码的蛋白为碱性亲水性蛋白,其中前23个氨基酸可能为信号肽,IFN-γ编码蛋白保守结构域为IFN-γ超家族,且存在跨膜结构,其中1-6位氨基酸为胞内区域,7-28位氨基酸为跨膜区域,29-167位氨基酸为胞外区域;IFN-γ编码蛋白二级结构主要以α-螺旋(58.08%)和无规则卷曲(33.53%)为主,存在5个潜在的B细胞抗原表位,3个潜在的N-糖基化位点;分子进化分析显示,东北虎IFN-γ与GenBank上发表的东北虎、非洲狮、金钱豹、美洲狮、猎豹、家猫、加拿大猞猁、野猪等的核苷酸相似性为80.4%~99.8%,氨基酸相似性为70.5%~100%,东北虎IFN-γ与非洲狮、金钱豹亲缘关系最近,美洲狮、猎豹、家猫、猞猁次之,野猪最远。通过合成改造后的东北虎IFN-γ基因,构建能表达IFN-γ蛋白的重组质粒pPIC9K-IFN-γ,将其导入高效表达系统-毕赤酵母中进行诱导表达,经SDS-PAGE分析,表达蛋白的分子量约17.8 ku,与预期大小相符,表明东北虎IFN-γ成功表达。     

Effects of ganmao shuangjie heji on the inflammatory damage in the lungs of mice infected by IV FM1

AIM: To establish the influenza infected mouse model and study the anti-inflammatory effect of ganmao shuangjie heji. METHODS: IV FM1 infected mice were used as the animal model. The changes of pathology and the cytokine TNF-α, IFN-γ and IL-10 in the lung were observed by HE staining and ELISA (double antibody sandwich enzyme linked immunosorbent assay) after ganmao shuangjie heji treatment. RESULTS: After infected by influenza virus, severe interstitial pneumonia was induced in the model group. Mild interstitial pneumonia was observed in ganmao shuangjie heji treated group. The protein expressions of cytokine TNF-α, IFN-γ and IL-10 were higher in model group than those in the control group. The protein expressions of TNF-α and IFN-γ in ganmao shuangjie heji treated group decreased and IL-10 expression increased significantly compared with model group. CONCLUSION: Ganmao shuangjie heji decreases the expressions of TNF-α and IFN-γ, and increases the expression of IL-10, thus, alleviates inflammatory injury. The clinical application of this medicine can shorten the course of disease.     

乙酸浓度对奶牛乳腺上皮细胞甘油三酯含量及瘦素和过氧化物酶增殖物激活受体γ基因表达量的影响

本试验旨在研究乙酸浓度对奶牛乳腺上皮细胞甘油三酯(TAG)含量及瘦素(leptin)和过氧化物酶增殖物激活受体γ(PPARγ)基因表达量的影响。将传至第3代的奶牛乳腺上皮细胞以适宜的密度培养48 h后,随机分为6个处理,各处理分别采用乙酸浓度为0(对照组)、4、6、8、10和12 mmol/L的培养液。置于37℃5%CO2培养箱继续培养48 h。收集细胞,观测脂滴形成状况,测定TAG含量及leptin和PPARγ基因表达量。结果表明:随乙酸浓度的增加,培养的奶牛脂肪前体细胞脂滴形成增多;随乙酸浓度的增加,TAG含量和PPARγ基因表达量均呈显著的线性或二次曲线增加(P<0.05),且以乙酸浓度为10和12 mmol/L时效果较好;一定浓度的乙酸可上调leptin基因表达量,尤以浓度为8、10 mmol/L时上调作用较好。结果提示,乙酸对奶牛乳腺上皮细胞内脂滴的形成、TAG的积累、leptin和PPARγ基因的表达有显著的促进作用。本试验条件下,乙酸浓度为10～12 mmol/L时能较好地促进PPARγ基因的表达,浓度为8～10 mmol/L时能较好地促进leptin基因的表达。     

绵羊磷脂酶C-γ1对绵羊卵母细胞体外成熟的影响

卵母细胞成熟率可作为衡量体外培养卵母细胞质量和发育能力的指标。本研究旨在探讨PLC-γ1是否参与了绵羊卵母细胞的体外成熟及其影响。将绵羊卵母细胞在含有不同浓度的U73122（PLC抑制剂））及m-3M3FBS （PLC激活剂）的成熟培养液中进行培养,每个浓度150个细胞,重复试验3次,统计细胞成熟率、卵裂率及桑葚胚率以供筛选出PLC-γ1的最佳抑制及促进浓度。qPCR检测0.5 μmol·L^-1 U73122及0.5 μmol·L^-1 m-3M3FBS处理后48 h的卵母细胞中PLC-γ1、BAK、BAX、CASP3、CASP8、P53、BCL6基因mRNA水平表达情况,Western blot检测0.5 μmol·L^-1 U73122及0.5 μmol·L^-1 m-3M3FBS处理后48 h的卵母细胞中PLC-γ1、BAK、BAX、CASP3、CASP8、P53、BCL6蛋白的表达情况。每个浓度150个细胞,重复试验3次。在绵羊卵母细胞中,0.5 μmol·L^-1 U73122是促进成熟的最佳浓度,0.5 μmol·L^-1 m-3M3FBS是抑制成熟的最佳浓度。0.5 μmol·L^-1 U73122处理组卵裂率和囊胚率均显著低于对照组。0.5 μmol·L^-1 m-3M3FBS处理组卵裂率和囊胚率均显著高于对照组。通过qPCR检测结果显示,与对照组相比,U73122组中BAK、BAX、CASP3、CASP8、P53基因mRNA表达量显著上升,PLC-γ1、BCL6基因mRNA表达量显著下降;m-3M3FBS组则与之相反。Western blot结果显示,与对照组相比,U73122组中BAK、BAX、CASP8蛋白表达量显著增多,PLC-γ1、BCL6蛋白表达量显著减少,P53和CASP3蛋白表达量无明显变化。m-3M3FBS组中PLC-γ1、BCL6蛋白表达量显著增多,BAX、CASP3、P53蛋白表达量显著减少,BAK、CASP8蛋白表达量无明显变化。综上所述,本研究表明PLC-γ1在绵羊卵母细胞的体外成熟培养中发挥重要作用,其可调节卵母细胞的成熟以及早期胚胎的发育能力。     

PLCγ1基因对绵羊早期胚胎体外发育的影响

旨在探究磷脂酶Cγ1（phospholipase Cgamma 1,PLCγ1）基因对绵羊早期胚胎体外发育的影响,为揭示PLCγ1基因在早期胚胎发育过程中的作用机制奠定基础。本研究选取包裹3层以上颗粒细胞的绵羊卵母细胞为试验材料,体外培养成熟后将其分为两组,利用显微注射技术将PLCγ1基因导入MⅡ期（减数第二次分裂中期）卵母细胞（n=127）中为试验组,以显微注射空载体pcDNA3.1-EGFP作为对照组（n=120）,经48 h后统计其卵裂率;并利用Ca²⁺探针Rhod 2-AM监测MⅡ卵母细胞和显微注射后16、48、72、96、120 h时各时期卵母细胞以及早期胚胎内Ca²⁺波动。结果显示,PLCγ1基因显微注射后卵母细胞被激活,卵裂率为（（19.67±0.2）%,P<0.01）,桑椹胚发育率为（（9.00±0.17）%,P<0.05）;PLCγ1基因注射后,卵母细胞和早期胚胎不同发育时期有Ca²⁺浓度变化,可观察发现Ca²⁺主要分布在胞质中且注射48 h时可达到峰值（P<0.01）。结果表明,PLCγ1基因可以引起绵羊卵母细胞发育过程中发生Ca²⁺振荡,启动其卵裂,并促使早期胚胎的继续发育。     

观赏植物组培与辐射结合育种研究进展

综合分析了组培与辐射结合育种方式的优点,对这种复合育种方式在观赏植物上的应用进行了分析归纳。总结出组培与辐射结合育种体系的基本技术路线,指出不同植物、不同材料的辐射效果的差异,并列出一些供育种者参考辐射剂量。最后分析了以后的研究方向。     

农杆菌介导含硫氨基酸γ-zein转化菊苣的初步研究

含硫氨基酸具有动物营养与免疫相关的重要生理功能,为提高菊苣中含硫氨基酸含量,采用根癌农杆菌介导法将玉米种子贮藏蛋白含硫氨基酸基因γ-zein和绿色荧光蛋白GFP融合基因转入到菊苣无菌苗叶片中,经过共培养、潮霉素抗性筛选、分化、再生和炼苗,得到抗性植株。对抗性植株进行PCR、PCR-Southern、斑点杂交和RTPCR分析,结果表明,外源目的基因已经整合到菊苣基因组中并且得到了表达,为提高菊苣含硫氨基酸含量,改善其品质奠定了基础。     

Effect of Nutrient Restriction and Compensation on Weight,Serum Indexes and Gene Expression of Chest Fat Adipokines in Mongolian Lamb

To study the effect of nutrient restriction and compensation on weight,serum indexes and gene expression of chest fat adipokine in Mongolian lamb,thirty male lambs were randomly divided into control group (20.26 kg± 3.63 kg) and feed restricted group (21.29 kg±3.00 kg).The experiment lasted for 120 days,including 60 days for feed restriction and 60 days for compensatory feed.At the end of each stage,five lambs from each group were slaughtered and chest fat was collected.Serum concentration of triglycerides,non-esterified fatty acids (NEFA),total cholesterol were measured.Quantitative Real-time PCR were used to determine Adiponectin,Visfatin,PPAR-γ and ACC genes expression.The results showed that:①The lamb body weight of restricted group kept maintain level and that of control group showed a increase tendency.During the compensatory period,the average daily gain (ADG) of restricted group was significantly higher than control group (P< 0.05).②During the restriction period,serum triglycerides concentration of lambs in restricted group was significantly decreased at day 30 and 60 (P< 0.05),and NEFA concentration was significantly increased at day 30 (P< 0.05).After 60 days of nutritional compensation,all serum parameters we tested were not significantly different between restrict group and control group (P >0.05).③During the restriction period,PPAR-γ and ACC gene expression in restricted group were significantly lower than that of control group (P< 0.05),while no significant differences were found during the compensation period (P>0.05).Adiponectin and Visfatin genes expression in chest fat was not significantly changed throughout the experiment (P >0.05).We could know that nutritional restriction reduced fatty acid synthesis and adipocyte differentiation ability,adipose tissue was mobilized energy metabolism,and total cholesterol was recoveried growth on the compensation.However,the ACC and PPAR-γ genes in restricted group were significantly decreased with the body weight increased rapidly during the compensatory stage.Adiponectin and Visfatin were not significantly different between restricted and compensatory stages.