氧化型低密度脂蛋白对人脐静脉内皮祖细胞增殖、凋亡及bcl-2表达的影响

目的探讨氧化型低密度脂蛋白对人脐静脉内皮祖细胞增殖、凋亡及bcl-2表达的影响。方法密度梯度离心法获取人脐静脉血单个核细胞,培养7天后,收集贴壁细胞并加入不同浓度氧化型低密度脂蛋白(分别为5、10及20mg/L)干预48h。MTT法检测氧化型低密度脂蛋白对内皮祖细胞增殖能力的影响,流式细胞仪检测氧化型低密度脂蛋白对细胞凋亡率的影响,逆转录聚台酶链反应检测bcl-2mRNA的表达,免疫细胞化学法检测bcl-2蛋白的表达。结果氧化型低密度脂蛋白呈浓度依赖性抑制内皮祖细胞增殖,诱导内皮祖细胞凋亡(P<0.01);基础状态下内皮祖细胞表达bcl-2mRNA及蛋白,氧化型低密度脂蛋白处理能抑制其表达,并存在量效关系(P<0.05)。结论氧化型低密度脂蛋白通过下调bcl-2的表达诱导内皮祖细胞凋亡、抑制内皮祖细胞增殖,这可能影响血管内皮的修复及新生血管的形成。     

Evolutionary Patterns of the Proviral gp90 V3 to V5 Regions of Equine Infectious Anemia Virus Associated with Immune Selection in Progressors and Nonprogressors

The aim of this study was to determine the genomic evolutionary pattern of virulent equine infectious anemia virus (EIAV)during persistent infection.The evolutionary dynamics of proviral genomes were examined by challenging an EIAV seronegative equine (pony 1) and three EIAV vaccinated equines (ponies 4,7,and 8) with the Chinese virulent strain EIAVL.Ponies I and 7 succumbed to disease and were called progressors,while ponies 4 and 8 lacked clinical symptoms and were considered nonprogressors.Sequences spanning the V3,V4,and V5 hyper-variable regions of the EIAV-L envelope gp90 gene were sequenced from each pony as evolutionary markers of the provirus.The proviral genome of the EIAV-Linoculum evolved during persistent infection and displayed different patterns between EIA progressors and nonprogressors.Inoculum-like variants were isolated from nonprogressors during persistent infection,but only from progressors during acute infection.Variant mutations from nonprogressors were dispersed throughout the sequenced region,while those from progressors were predominantly localized to V3.Humoral immunity and virus variant population selection analyses indicated that immune selection was positive in chronically infected progressors and weak in nonprogressors.In-frame stop codons were frequently localized to a defect "hot spot".The high number of defective variants in nonprogressors may promote disease survival.     

HIV/AIDS患者外周血中4种细胞因子IL-2、IL-4、IL-15和IL-21表达水平及其临床意义研究

目的 观察外周血 4 种细胞因子（IL-2、IL-4、IL-15及IL-21）在HIV感染者/AIDS患者（HIV/AIDS）和正常人表达水平的区别及其与HIV RNA病毒载量和CD3⁺、CD4⁺细胞数量的相关性,分析其与HIV感染的关系和临床意义。方法 40例未经抗病毒治疗的HIV/AIDS 和31名正常人,采用ELISA法检测外周血4种细胞因子（IL-2、IL-4、IL-15及IL-21）,使用流式细胞计数法检测外周血CD3⁺、CD4⁺ T 淋巴细胞数量,实时聚合酶链反应测定血清中HIV RNA水平。结果 未经抗病毒治疗的HIV/AIDS外周血IL-2、IL-15及IL-21水平低于正常人,而IL-4水平高于正常人（P<0.05）。CD3⁺、CD4⁺细胞数量和各细胞因子水平显示出一定的相关关系,IL-2和IL-15水平均与HIV-1 RNA载量呈负相关。结论 提示外周血4种细胞因子都可能参与了HIV感染的致病机制,并与HIV RNA病毒载量及CD3⁺、CD4⁺细胞数量相关。     

中国马传染性贫血病毒驴强毒株感染性分子克隆的构建

 马传染性贫血病毒（equine infectious anemia virus, EIAV）驴强毒株DV是一株经过驴体传代获得的而具有超强毒力的毒株，对马和驴均可100％致死。用PCR方法分段扩增了DV其前病毒基因，将包含全基因片段的3个基因克隆以限制性内切酶消化后顺次连接克隆到pLG338上，命名为pD70344。将此克隆体外转染驴胎皮肤细胞和驴白细胞，连续盲传3代并以反转录酶活性测定，RT-PCR鉴定其病毒活性，透射电镜观察发现细胞培养物中存在大量典型病毒粒子，证明获得了1株具有感染性的EIAV病毒粒子，命名为pD70344V。经序列测定确认了本试验首次构建了1株完全来源于EIAV强毒基因的感染性分子克隆，将此克隆病毒接种驴，可以引起典型的马传贫症状并导致试验动物死亡。该分子克隆的建立为进一步考察病毒的毒力与基因的关系提供了良好的平台。     

马传染性贫血病毒在慢性感染体内的抗原变异

应用在日本发离的马传染性贫血病毒（ＥＩＡＶ）强毒株Ｐ３３７－Ｖ７０实验感染了１匹健康马，经２次感染后，这匹实验感染并未呈现临床症状。从这匹马的外周和肝组织获得了覆盖马传染性贫血病毒ｇｐ９０基因主要中和抗原功能区（ＰＮＤ）的前病毒序列。通过对这些序列的比较分析发现，一些序列在ＰＮＤ功能区含有核苷酸的插入变异。     

Isolation and transmission of human retrovirus (human t-cell leukemia virus)

Nine new isolates of human T-cell leukemia-lymphoma virus (HTLV) were obtained from cells of seven patients with malignancies of mature T cells and from two clinically normal relatives of a T-cell leukemia patient. These people were from the United States, Israel, the West Indies, and Japan. The virus was detected in the fresh T cells and was isolated from the established T-cell lines. Each isolate is closely related to the first HTLV isolate, and all the new HTLV isolates were transmitted into normal human T cells obtained from the umbilical cord blood of newborns.     

NOV (CCN3) functions as a regulator of human hematopoietic stem or progenitor cells

Clinically successful hematopoietic cell transplantation is dependent on hematopoietic stem and progenitor cells. Here we identify the matricellular protein Nephroblastoma Overexpressed (Nov, CCN3) as being essential for their functional integrity. Nov expression is restricted to the primitive (CD34) compartments of umbilical vein cord blood, and its knockdown in these cells by lentivirus-mediated RNA interference abrogates their function in vitro and in vivo. Conversely, forced expression of Nov and addition of recombinant Nov protein both enhance primitive stem and/or progenitor activity. Taken together, our results identify Nov (CCN3) as a regulator of human hematopoietic stem or progenitor cells.     

Effects of Bupleurum extract on blood metabolism,antioxidant status and immune function in heat-stressed dairy cows

This experiment was conducted to evaluate the effects of Bupleurum extract(BE) on blood metabolites, antioxidant status, and immune function in dairy cows under heat stress. Forty lactating Holstein cows were randomly assigned to 1 of 4 treatments. The treatments consisted of 0, 0.25, 0.5, and 1.0 g of BE kg–1 dry matter. Supplementation with BE decreased(P0.05) blood urea nitrogen(BUN) contents and increased blood total protein(TP) and albumin(ALB) levels compared with control cows, but it had no effects(P0.05) on blood glucose(GLU), nonesterified fatty acid(NEFA), total triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), and high density lipoprotein cholesterol(HDL-C). Compared with control cows, cows fed BE had higher(P0.05) superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activity. However, supplementation with BE had no effect(P0.05) on total antioxidant capacity(T-AOC) or malondialdehyde(MDA) levels. The immunoglobulin(Ig) A and G contents increased(P0.05) in cows fed 0.25 or 0.5 g of BE kg–1. Interleukin(IL)-2 and IL-4 levels were higher(P0.05) in cows fed 0.5 and 1.0 g of BE kg–1, and IL-6 was significantly elevated(P0.05) in cows fed 0.5 g of BE kg–1. There were no treatment effects(P0.05) on the CD4+ and CD8+ T lymphocyte ratios, CD4+/CD8+ ratio, or tumor necrosis factor-α(TNF-α) level among the groups. These findings suggest that BE supplementation may improve protein metabolism, in addition to enhancing antioxidant activity and immune function in heat-stressed dairy cows.     

固始鸡脐带间充质干细胞原代培养及其诱导分化潜能研究

为了研究固始鸡脐带间充质干细胞（umbilical cord mesenchymal stem cells,UCMSCs）的生物学特性和诱导分化,对其进行原代培养及定向诱导分化研究。选取14日龄固始鸡鸡胚的脐带组织,剥离脐带动、静脉,将华通胶部分剪成1 mm³大小,采用胶原酶消化法分离细胞并进行原代培养,观察细胞形态,绘制P3、P9、P15代生长曲线并测定群体倍增时间及克隆形成能力。结果表明,固始鸡UCMSCs形态为长梭形,生长趋势符合Logistic生长曲线规律,呈典型的S形;免疫荧光和RT-PCR结果显示,固始鸡UCMSCs表达CD29、CD44、CD73、CD90、CD166等间充质干细胞表面标记物基因,但不表达原始造血祖细胞标志物CD34和泛白细胞标志物CD45;经特异性染色和RT-PCR表明,体外诱导固始鸡UCMSCs可分化为脂肪、成骨和成软骨细胞。从固始鸡脐带华通胶中分离所获得的UCMSCs具有良好的体外增殖能力和分化潜能,与从其他物种体内分离的UCMSCs具有相似的生物学特征,它们的多项分化能力预示着细胞移植的应用前景,可为再生医学和组织工程学提供新的潜在种子细胞。     

多普勒超声脐动脉血流图在产前监测中的应用

目的 :评价脐动脉超声血流图在产前监测中的作用。方法 :孕妇 2 6 0例 (孕 37～ 42周 )用多普勒超声行脐动脉血流图检查并计算 S/ D值 ,胎心无负荷试验 (NST) ,检查结果与分娩后新生儿出生体重、羊水污染、脐带绕颈或过短、新生儿窒息等进行比较。结果 :S/ D值异常组 6 9例 ,其中分娩低体重儿 8例 (11.6 % )、发生羊水污染 30例 (4 3.5 % )、脐带缠绕或过短 2 0例 (9.0 % )、新生儿窒息 12例 (17.4% ) ,均较 S/ D值正常组明显增高 (P<0 .0 1)。结论 :超声脐动脉血流图在判断宫内胎儿变化中起着重要作用 ,与 NST联合应用对围产儿预后能作出早期诊断 ,能提高围产期保健质量。     

Human monocytic cell lines derived from cord leukocytes by co-cultivation with irradiated CM-S cells

The CM-S cell line was established from the bone marrow of a child with congenital hypoplastic anemia and resembles its monocyte-macrophage lineage. Lethally x-irradiated CM-S cells from various passages and clones, representing different stages in the progression of the transformed growth phenotype, were tested for their ability to affect the survival and proliferation of normal human cord or adult blood leukocytes in co-culture. One clone, CM-SM, which is tumorigenic in athymic mice, consistently immortalized umbilical cord mononuclear cells but did not immortalize adult peripheral blood leukocytes. Six autonomous monocyte-like diploid cell lines were obtained and all were found to be of cord origin. Three lines were tumorigenic in athymic mice. Attempts to immortalize human leukocytes with cell-free supernatants from CM-S cells were unsuccessful.     

A double-layer model for improving the estimation of wheat canopy nitrogen content from unmanned aerial vehicle multispectral imagery

The accurate and rapid estimation of canopy nitrogen content (CNC) in crops is the key to optimizing in-season nitrogen fertilizer application in precision agriculture. However, the determination of CNC from field sampling data for leaf area index (LAI), canopy photosynthetic pigments (CPP; including chlorophyll a, chlorophyll b and carotenoids) and leaf nitrogen concentration (LNC) can be time-consuming and costly. Here we evaluated the use of high-precision unmanned aerial vehicle (UAV) multispectral imagery for estimating the LAI, CPP and CNC of winter wheat over the whole growth period. A total of 23 spectral features (SFs; five original spectrum bands, 17 vegetation indices and the gray scale of the RGB image) and eight texture features (TFs; contrast, entropy, variance, mean, homogeneity, dissimilarity, second moment, and correlation) were selected as inputs for the models. Six machine learning methods, i.e., multiple stepwise regression (MSR), support vector regression (SVR), gradient boosting decision tree (GBDT), Gaussian process regression (GPR), back propagation neural network (BPNN) and radial basis function neural network (RBFNN), were compared for the retrieval of winter wheat LAI, CPP and CNC values, and a double-layer model was proposed for estimating CNC based on LAI and CPP. The results showed that the inversion of winter wheat LAI, CPP and CNC by the combination of SFs+TFs greatly improved the estimation accuracy compared with that by using only the SFs. The RBFNN and BPNN models outperformed the other machine learning models in estimating winter wheat LAI, CPP and CNC. The proposed double-layer models (R²=0.67–0.89, RMSE=13.63–23.71 mg g^–1, MAE=10.75–17.59 mg g^–1) performed better than the direct inversion models (R²=0.61–0.80, RMSE=18.01–25.12 mg g^–1, MAE=12.96–18.88 mg g^–1) in estimating winter wheat CNC. The best winter wheat CNC accuracy was obtained by the double-layer RBFNN model with SFs+TFs as inputs (R²=0.89, RMSE=13.63 mg g^–1, MAE=10.75 mg g^–1). The results of this study can provide guidance for the accurate and rapid determination of winter wheat canopy nitrogen content in the field.     

In-vitro assessment for the control of Fusarium species using a lactic acid bacterium isolated from yellow pitahaya(Selenicereus megalanthus(K. Schum. Ex Vaupel Moran))

The fungistatic activity of a lactic acid bacterium, which had been isolated from yellow pitahaya cultures, against fungi associated with basal rot(Fusarium oxysporum and Fusarium fujikuroi) was measured in the present study. Its activity was assessed in three fractions: fermented(S1), metabolic products(S2), and biomass(S3), using two fermentation substrates: Man Rogosa Sharpe agar(MRS) and potato dextrose agar(PDA). The bacterium was molecularly identified as Lactobacillus plantarum. S3 reduced F. fujikuroi growth by 100% over 48 h of fermentation, which occurred during the stationary phase of bacterial growth. The three fractions' fungistatic activity against F. fujikuroi depended on the substrate employed. The fermentation kinetic parameters for L. plantarum indicated that its specific growth rate was 0.46 h~(–1), with 93.63% substrate consumption, 0.045 kg kg~(–1) cell yield, and 0.54 kg kg~(–1) product yield. The kinetic parameters calculated will allow for bacteria production scaling. These in-vitro trials reveal L. plantarum's possible application as a biocontrol agent for diseases associated with Fusarium. However, further ex-vivo and in-vivo researches are required to demonstrate its behavior in crops.     

Nitrogen nutrition diagnosis for cotton under mulched drip irrigation using unmanned aerial vehicle multispectral images

Remote sensing has been increasingly used for precision nitrogen management to assess the plant nitrogen status in a spatial and real-time manner. The nitrogen nutrition index (NNI) can quantitatively describe the nitrogen status of crops. Nevertheless, the NNI diagnosis for cotton with unmanned aerial vehicle (UAV) multispectral images has not been evaluated yet. This study aimed to evaluate the performance of three machine learning models, i.e., support vector machine (SVM), back propagation neural network (BPNN), and extreme gradient boosting (XGB) for predicting canopy nitrogen weight and NNI of cotton over the whole growing season from UAV images. The results indicated that the models performed better when the top 15 vegetation indices were used as input variables based on their correlation ranking with nitrogen weight and NNI. The XGB model performed the best among the three models in predicting nitrogen weight. The prediction accuracy of nitrogen weight at the upper half-leaf level (R²=0.89, RMSE=0.68 g m^–2, RE=14.62% for calibration and R²=0.83, RMSE=1.08 g m^–2, RE=19.71% for validation) was much better than that at the all-leaf level (R²=0.73, RMSE=2.20 g m^–2, RE=26.70% for calibration and R²=0.70, RMSE=2.48 g m^–2, RE=31.49% for validation) and at the plant level (R²=0.66, RMSE=4.46 g m^–2, RE=30.96% for calibration and R²=0.63, RMSE=3.69 g m^–2, RE=24.81% for validation). Similarly, the XGB model (R²=0.65, RMSE=0.09, RE=8.59% for calibration and R²=0.63, RMSE=0.09, RE=8.87% for validation) also outperformed the SVM model (R²=0.62, RMSE=0.10, RE=7.92% for calibration and R²=0.60, RMSE=0.09, RE=8.03% for validation) and BPNN model (R²=0.64, RMSE=0.09, RE=9.24% for calibration and R²=0.62, RMSE=0.09, RE=8.38% for validation) in predicting NNI. The NNI predictive map generated from the optimal XGB model can intuitively diagnose the spatial distribution and dynamics of nitrogen nutrition in cotton fields, which can help farmers implement precise cotton nitrogen management in a timely and accurate manner.     

副猪嗜血杆菌对猪气管上皮细胞的黏附作用及其黏附模型的建立

为了探讨副猪嗜血杆菌(Haemophilus parasuis,Hps)与原代培养的猪气管黏膜上皮细胞的相互作用,应用气管支气管结扎灌注酶冷消化法分离气管黏膜上皮细胞,于胶原覆盖的盖玻片上培养,使上皮细胞分化成假复层黏膜纤毛上皮细胞,然后以对数生长期不同Hps含量的菌液感染上皮细胞和猪脐静脉血管内皮细胞,分别作用1、2、34、h后,通过革兰氏染色法观察Hps对细胞的黏附情况。结果表明,Hps可黏附在上皮细胞和内皮细胞表面;Hps对气管上皮细胞有毒性作用,致细胞变性坏死和凋亡;Hps对细胞的黏附力与其在菌液中含量和作用时间有关,以菌落形成单位计,当含量为1.0×108mL-1且感染4 h后,黏附效果最好。高倍显微镜下可见大量Hps黏附于猪气管上皮细胞,只有个别Hps黏附于猪血管内皮细胞表面。说明Hps对猪气管黏膜上皮细胞的黏附能力远强于猪血管内皮细胞,可以利用猪气管黏膜上皮细胞建立副猪嗜血杆菌的黏附细胞模型。     

Organic and inorganic fertilization influenced on yield and quality of sugar beet genotypes

The objective of this study was to evaluate the effects of organic and inorganic fertilizers on yield and quality of sugar beet genotypes (Beta vulgaris L.). Therefore a field trial was carried out at New Developmental Farm of The University Agriculture, Peshawar, Pakistan during winter 2012–13. The field experiment was layout in randomized complete block design with split plot arrangement having three replications. Fertilizers treatments (control, higo organic plus (composted manure, it contains N 2%, P 3%, K 3%, organic matter 40%, organic carbon 11%, Zn 145 mg kg^–1, Cu 56 mg kg^–1, Fe 380 mg kg^–1 and Mg 228 mg kg^–1), maxicrop sea gold (extract of sea weeds i.e., Sargasssum, Laminara Polysaccharide and Ascophyllum Nodosum), farm yard manure, NP (90: 60 kg ha^–1), NP (120: 90 kg ha^–1) and NP (150: 120 kg ha^–1) were allotted to main plots while genotypes (Sandrina, Serenada and Kawe Terma) to the sub plots. Farm yard manure (10 t ha^–1) and higo organic Plus (5 t ha^–1) was incorporated in the soil before seed bed preparation. Maxicrop sea gold (5 lit ha^–1) was sprayed after the emergence of the crop. All phosphorus was applied @ of 60, 90 and 120 kg ha^–1 at the time of sowing while nitrogen @ of 90, 120 and 150 kg ha^–1 in two splits/3 of the dose was applied at the time of sowing while the remaing 1/3 of the dose was applied before earthen up. Plots treated with application of NP ratio 120: 90 kg ha^–1 produced maximum beet yield (76.4 t ha^–1), sugar yield (11.1 t ha^–1), Pol (polarizable sugar) percentage (14.67%) and more economic return (Rs.234 Thousand ha^–1) as compared to control plots. Sugar beet genotype Serenada had significantly produced maximum beet yield (55.5 t ha^–1), sugar yield (7.9 t ha^–1), pol (polarizable sugar) percentage (14.60%), brix percentage (14.60%) and more economic return (158) as compared to other genotypes. It was concluded from the above results that sugar beet genotype Serenada treated with NP ratio 120: 90 kg ha^–1 for improved sugar beet productivity and quality therefore it is recommended for general practice in agro-climatic conditions of Peshawar valley.     

Cultivar selection can increase yield potential and resource use efficiency of spring maize to adapt to climate change in Northeast China

Northeast China (NEC) is one of the major maize production areas in China.Agro-climatic resources have obviously changed,which will seriously affect crop growth and development in this region.It is important to investigate the contribution of climate change adaptation measures to the yield and resource use efficiency to improve our understanding of how we can effectively ensure high yield and high efficiency in the future.In this study,we divided the study area into five accumulated temperature zones (ATZs) based on growing degree days (GDD).Based on the meteorological data,maize data (from agrometeorological stations) and the validated APSIM-Maize Model,we first investigated the spatial distributions and temporal trends of maize potential yield of actual planted cultivars,and revealed the radiation use efficiency (RUE) and heat resource use efficiency (HUE) from 1981 to 2017.Then according to the potential growing seasons and actual growing seasons,we identified the utilization percentages of radiation (P_R) resource and heat resource (P_H) for each ATZ under potential production from 1981 to 2017.Finally,we quantified the contributions of cultivar changings to yield,P_R and P_H of maize.The results showed that during the past 37 years,the estimated mean potential yield of actual planted cultivars was 13 649 kg ha~(–1),ranged from 11 205 to 15 257 kg ha~(–1),and increased by 140 kg ha~(–1) per decade.For potential production,the mean values of RUE and HUE for the actual planted maize cultivars were 1.22 g MJ~(–1) and 8.58 kg (℃ d)~(–1) ha~(–1).RUE showed an increasing tendency,while HUE showed a decreasing tendency.The lengths of the potential growing season and actual growing season were 158 and 123 d,and increased by 2 and 1 d per decade.P_R and P_H under potential production were 82 and 86%,respectively and showed a decreasing tendency during the past 37 years.This indicates that actual planted cultivars failed to make full use of climate resources.However,results from the adaptation assessments indicate that,adoption of cultivars with growing season increased by 2–11 d among ATZs caused increase in yield,P_R and P_H of 0.6–1.7%,1.1–7.6% and 1.5–8.9%,respectively.Therefore,introduction of cultivars with longer growing season can effectively increase the radiation and heat utilization percentages and potential yield.     

Ameliorating Nickel Stress by Jasmonic Acid Treatment in <Emphasis Type="Italic">Zea mays</Emphasis> L.

Plants need various micro and macronutrients for their growth and metabolism. Of these nickel (Ni) plays a role of micronutrient but causes adverse effects when present above optimum level. To combat such situations plants possess different growth regulators. Jasmonates are a class of plant growth regulators modulating various growth and physiological responses in plants. Aiming to evaluate the effect of nickel and jasmonic acid on growth and antioxidant enzymes Superoxide distmutase (SOD), Catalse (CAT), Ascorbate peroxidase (APX), Guaiacol peroxidase (GPX) and glutathione reductase (GR), seeds of Zea mays L. were sown in field after giving presowing treatment of NiSO₄ · 6H₂O (8 mM), jasmonic acid JA (10^–6, 10^–8 and 10^–10 M) alone and JA (10^–6, 10^–8 and 10^–10 M) in 1: 1 combination with NiSO₄ · 6H₂O (8 mM) in plant conservatory, Punjabi University, Patiala, Punjab, India. Plants of each treatment group were then subjected to spraying treatment with the respective treatment solutions at regular intervals of 22 days up to 66 days after sowing (DAS). The present findings signified reduced growth with Ni treatment but co-application of JA alleviated the effect of nickel through activity of antioxidant enzymes.