高蛋白高产大豆新品种石豆11号的选育

石豆11号是由石家庄市农林科学研究院以化诱446为母本,冀豆4号为父本经有性杂交选育而成。2014-2015年参加河北省夏播大豆区域试验,平均产量3 093.9 kg·hm~(-2),较对照冀豆12增产2.51%。2016年参加河北省夏播大豆生产试验,平均产量3 108.0 kg·hm~(-2),较对照冀豆12增产0.87%。2017年通过河北省农作物品种审定委员会审定,准予推广。石豆11号的主要特点是高蛋白(45.79%)、高产、抗倒伏,适宜在河北省中南部夏播种植。     

Antioxidant and anti‐inflammatory activities of Pinus radiata bark extract in salmonid cell lines

A fish meal supply shortage is limiting aquaculture development. Currently, plant‐based proteins, such as soya bean meal, are being used as an alternative protein source, despite that such a diet can adversely affect fish, such as by inducing an inflammatory response. A possible solution is to include dietary additives in farm diets to counteract negative effects. One such solution originates from pine bark extracts, which present bioactive properties. In this study, the antioxidant and anti‐inflammatory properties of Pinus radiata bark extracts were evaluated for the first time in a salmonid cell line. This extract chemically demonstrated antioxidant activity through 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH = 58.4 ± 1.1%) and ferric ion reducing antioxidant power (FRAP = 575 ± 17 mgEqFe(II)·g extract^?1) assays. Additionally, the extract showed high flavonoid and phenolic compound contents. Up to 100 mg mL^?1, the P. radiata extract showed no cytotoxicity in the CHSE‐214 salmonid embryo cell line. Moreover, the antioxidant activity of the extract (50 μg mL^?1) was evaluated by a dichlorofluorescein (DCFH) assay in the SHK‐1 salmon cell line challenged with an oxidant stimulus (H₂O₂), showing 58.9% activity. The extract also protected DNA from oxidative damage, as observed through a comet assay. When assessing anti‐inflammatory properties in an in vitro inflammation model, the extract significantly reduced the relative expression of the pro‐inflammatory cytokines interleukin‐1β (IL‐1β), tumour necrosis factor‐α (TNF‐α) and interleukin‐8 (IL‐8) and of the inducible cyclooxygenase‐2 (COX‐2) enzyme. These results suggest a potential application of P. radiata bark extract in functional foods in aquaculture.     

原花青素A-1体内外对猪与小鼠T淋巴细胞亚群的影响

为研究从碎米花杜鹃叶中分离得到的化合物原花青素A-1（Proanthocyanidin A-1，简称PAA-1）的免疫增强活性，初步从细胞水平上探讨其免疫增强机制。体外试验采用流式细胞术，检测PAA-1体外对小鼠脾淋巴细胞T细胞亚群CD4和CD8单阳性和双阳性T淋巴细胞亚群百分率及CD4+/CD8+比值；体内试验通过饲喂不同剂量的PAA-1后，检测试验猪外周血淋巴细胞CD4和CD8单阳性以及CD4+/CD8+比值。结果证明，体外试验中同阴性对照组相比，PAA-1能单独或协同Con A提升具CD4+、CD8+及CD4+CD8+表型的T淋巴细胞亚群百分率，提高CD4+/CD8+的比值；体内试验中饲喂中、高剂量的PAA-1的试验猪外周血中具有CD4+、CD8+表型的T淋巴细胞的百分率及CD4+/CD8+的比值均有显著提高。说明PAA-1可通过增加辅助性T淋巴细胞和细胞毒性T淋巴细胞数量、促进T淋巴细胞的成熟以及增加CD4+/CD8+的比值发挥增强细胞免疫的作用，为其进一步开发为新型免疫增强剂提供试验依据。     

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck, one of the most popular edible mushrooms, has attracted great attention due to its delicious taste and healthy properties. However, both its biological effects and the possible mechanism of action have not yet been known. We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck (MPE) against HepG2 human hepatocellular carcinoma cells. Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90% (P<0.01). The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax, Bcl-2 and cleaved caspase-3. On the other hand, MPE might trigger cell cycle arrest at G₀/G₁/S phases by managing p21, Cyclin D1, cyclin-dependent kinases-4 (CDK4) and proliferating cell nuclear antigen (PCNA). Additionally, MPE downregulated TRAF-2 and p-p53, while upregulated p-ASK1 and p-p38. Therefore, it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

抗条纹叶枯病基因STV11的功能性分子标记建立及粳型抗源筛选

条纹叶枯病是水稻(Oryza sativa L.)上最严重的病害之一。STV11是从籼稻Kasalath中鉴定的条纹叶枯病抗性基因,根据Kasalath型条纹叶枯病基因STV11 KAS6个碱基缺失的功能性多态性序列差异,设计基于PCR的功能性分子标记qstv11。利用此标记对299份试验材料[5份抗病品种、5份感病品种、41份辽宁省育种材料、17份日本粳型材料、142份秋光/七山占重组自交系(群体A)、85份笹锦/Habataki//笹锦回交重组自交系(群体B)、群体亲本4份]进行基因型分析并进行抗病性鉴定。结果表明,qstv11能够较好区分抗感基因型,而且在测试材料中与条纹叶枯抗性整体呈现共分离。41份辽宁材料中鉴定出含有STV11-R(抗)材料1份;日本粳型材料该位点均为STV11-S(感);群体A和B中分别筛选出粳稻背景STV11-R型材料24份和6份。     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.     

猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

兰州百合连作土壤水浸液自毒作用研究

【目的】探讨兰州百合(Lilium davidii var.unicolor salisb)连作土壤水浸液对自身幼苗生长的障碍因素及其作用机制。【方法】设置蒸馏水(CK)及正茬、连作2年、连作4年兰州百合根际土壤水浸液各50,100,200,300 mg/mL,共13个处理,以兰州百合种球为受试对象,测定不同条件下兰州百合幼苗生长、抗氧化酶活性、渗透调节物质及细胞膜透性的变化,并利用GC-MS技术分析各处理土壤中存在的主要自毒物质。【结果】正茬、连作2年及连作4年兰州百合根际土壤水浸液对兰州百合幼苗的生长均存在"低促高抑"现象,且抑制作用随连作年限的延长而增强。随水浸液质量浓度的增加,兰州百合幼苗CAT和SOD活性逐渐升高,POD活性先上升后下降,MDA含量、相对电导率、脯氨酸及可溶性糖含量的变化呈不断上升趋势,当水浸液质量浓度上升至300 mg/mL时,3个处理中各指标的上升或下降程度与对照相比均达显著差异水平(P0.05)。兰州百合正茬、连作2年及连作4年根际土壤中分别鉴定出9,15和17种化合物,主要包括2,3-丁二醇、1,2,3-三甲苯、邻苯二甲酸二丁酯、对苯二甲酸二辛酯、抗氧剂2246等化合物,其中大部分为自毒物质。【结论】兰州百合正茬、连作2年及连作4年根际土壤水浸液质量浓度达到300 mg/mL时,对其幼苗的生长会产生显著抑制作用。连作土壤中存在的自毒物质可以改变兰州百合植株体内的抗氧化酶活性,破坏细胞膜结构和功能,抑制兰州百合植株的生长,是导致兰州百合连作障碍发生的重要原因之一。