Physicochemical properties and development of wheat large and small starch granules during endosperm development

Wheat mature seeds have large, lenticular A-type starch granules, and small, spherical B-type and irregular C-type starch granules. During endosperm development, large amyloplasts came from proplastid, divided and increased in number through binary fission from 4 to 12 days after flowering (DAF). Large starch granules formed and developed in the large amyloplast. One large amyloplast had only one large starch granule. Small amyloplasts came from the protrusion of large amyloplast envelope, divided and increased in number through envelope protrusion after 12 DAF. B-type starch granules formed and developed in small amyloplast from 12 to 18 DAF, C-type starch granules formed and developed in small amyloplast after 18 DAF. Many B- and C-type starch granules might form and develop in one small amyloplast. The amyloplast envelopes were asynchronously degraded and starch granules released into cell matrix when amyloplasts were full of starch granules. Apparent amylose contents of large starch granules were higher than that of small starch granules, and increased with endosperm development. The swelling powers and crystallinity of large starch granule were lower than that of small starch granules, and decreased with endosperm development. Small starch granules displayed broader gelatinization temperature ranges than did large starch granules.     

表达T~+Pm保护性抗原的重组猪霍乱沙门氏菌C500株的构建及其生物学特性

【目的】本研究利用Asd+平衡致死系统构建表达巴氏杆菌毒素(Pasteurella multocida toxin,PMT)的重组猪霍乱沙门氏菌株,并对重组菌株的生物学特性进行比较研究。【方法和结果】通过基因克隆的方法构建表达PMT的重组质粒pYA-PmtC,再将其电转化减毒猪霍乱沙门氏菌C500的asd基因缺失株C501,构建口服活疫苗菌株C501(pYA-PmtC)。研究结果表明重组菌株C501(pYA-PmtC)的生化特性、血清型和生长速度与亲本菌株C500一致;在没有选择压力的条件下,C501(pYA-PmtC)能够稳定遗传重组质粒及其外源基因片段,并能稳定、高效、分泌性表达30.5kDa的外源保护性抗原rPmtC。C501(pYA-PmtC)腹腔感染BALB/c小鼠的LD50为8.5×106CFU,毒力稍低于C500(LD50为4.4×106CFU);口服接种C501(pYA-PmtC)和C500的所有仔猪未见任何发病症状,两者没有显著差别。【结论】本研究利用Asd+平衡致死系统的原理构建表达T+Pm保护性抗原重组猪霍乱沙门氏菌弱毒菌株C501(pYA-PmtC),为进一步开发猪萎缩性鼻炎-副伤寒的双价基因工程疫苗奠定基础。     

NEMO-binding domain peptide promotes osteoblast differentiation impaired by tumor necrosis factor alpha

Osteogenesis associated with persistent inflammation or infection exists in a broad range of conditions including rheumatoid arthritis and traumatic bone fracture. The poor outcomes of these conditions will benefit from more effective treatments. Here we investigated the molecular mechanisms and tested NEMO-binding domain peptide as a new approach of circumventing TNF-α inhibition of osteoblast differentiation. Our results showed: TNF-α markedly decreased BMP-2-induced alkaline phosphatase activity in the multipotent myoblast C2C12 cells in a dose dependent manner; stepwise experiments demonstrated that BMP-2-induced Smad1 activity was abrogated by addition of exogenous TNF-α or overexpression of NF-κB, and it was significantly elevated by overexpression of IκBα, an inhibitor of NF-κB; Western blotting showed that TNF-α markedly decreased the amount of phospho-Smad1 in BMP-2-activated C2C12 cells, but it did not alter Smad1 mRNA abundance as measured by real-time PCR; addition of a functional cell-permeable NEMO-binding domain (NBD) peptide antagonized NF-κB activity and ameliorated TNF-α inhibition of osteoblast differentiation. Taken together, our study reveals for the first time that NF-κB activation inhibits osteoblast differentiation by attenuating Smad1 activity and application of NBD peptide ameliorates this inhibitory effect. This could lead to new therapeutic drugs that circumvent the inflammatory inhibition of osteogenesis for treatment of traumatic open fractures with infection, rheumatoid arthritis and other bone loss disorders.     

VEGF-mediated angiogenesis stimulates neural stem cell proliferation and differentiation in the premature brain

This study investigated the effects of angiogenesis on the proliferation and differentiation of neural stem cells in the premature brain. We observed the changes in neurogenesis that followed the stimulation and inhibition of angiogenesis by altering vascular endothelial growth factor (VEGF) expression in a 3-day-old rat model. VEGF expression was overexpressed by adenovirus transfection and down-regulated by siRNA interference. Using immunofluorescence assays, Western blot analysis, and real-time PCR methods, we observed angiogenesis and the proliferation and differentiation of neural stem cells. Immunofluorescence assays showed that the number of vWF-positive areas peaked at day 7, and they were highest in the VEGF up-regulation group and lowest in the VEGF down-regulation group at every time point. The number of neural stem cells, neurons, astrocytes, and oligodendrocytes in the subventricular zone gradually increased over time in the VEGF up-regulation group. Among the three groups, the number of these cells was highest in the VEGF up-regulation group and lowest in the VEGF down-regulation group at the same time point. Western blot analysis and real-time PCR confirmed these results. These data suggest that angiogenesis may stimulate the proliferation of neural stem cells and differentiation into neurons, astrocytes, and oligodendrocytes in the premature brain.     

Genetic immunization based on the ubiquitin-fusion degradation pathway against Trypanosoma cruzi

Cytotoxic CD8⁺ T cells are particularly important to the development of protective immunity against the intracellular protozoan parasite, Trypanosoma cruzi, the etiological agent of Chagas disease. We have developed a new effective strategy of genetic immunization by activating CD8⁺ T cells through the ubiquitin-fusion degradation (UFD) pathway. We constructed expression plasmids encoding the amastigote surface protein-2 (ASP-2) of T. cruzi. To induce the UFD pathway, a chimeric gene encoding ubiquitin fused to ASP-2 (pUB-ASP-2) was constructed. Mice immunized with pUB-ASP-2 presented lower parasitemia and longer survival period, compared with mice immunized with pASP-2 alone. Depletion of CD8⁺ T cells abolished protection against T. cruzi in mice immunized with pUB-ASP-2 while depletion of CD4⁺ T cells did not influence the effective immunity. Mice deficient in LMP2 or LMP7, subunits of immunoproteasomes, were not able to develop protective immunity induced. These results suggest that ubiquitin-fused antigens expressed in antigen-presenting cells were effectively degraded via the UFD pathway, and subsequently activated CD8⁺ T cells. Consequently, immunization with pUB-ASP-2 was able to induce potent protective immunity against infection of T. cruzi.     

Regression Calibration in Semiparametric Accelerated Failure Time Models

Summary In large cohort studies, it often happens that some covariates are expensive to measure and hence only measured on a validation set. On the other hand, relatively cheap but error‐prone measurements of the covariates are available for all subjects. Regression calibration (RC) estimation method ( Prentice, 1982 , Biometrika 69 , 331–342) is a popular method for analyzing such data and has been applied to the Cox model by Wang et al. (1997, Biometrics 53 , 131–145) under normal measurement error and rare disease assumptions. In this article, we consider the RC estimation method for the semiparametric accelerated failure time model with covariates subject to measurement error. Asymptotic properties of the proposed method are investigated under a two‐phase sampling scheme for validation data that are selected via stratified random sampling, resulting in neither independent nor identically distributed observations. We show that the estimates converge to some well‐defined parameters. In particular, unbiased estimation is feasible under additive normal measurement error models for normal covariates and under Berkson error models. The proposed method performs well in finite‐sample simulation studies. We also apply the proposed method to a depression mortality study.     

TGF-α、TGF-β1在胰腺癌中的表达及临床意义

目的:研究TGF-α、TGF-β1与胰腺癌临床病理的关系.方法:免疫组织化学SABC法检测41例胰腺癌组织和12例正常胰腺组织中TGF-α、TGF-β1的表达情况,并分析其与病人的年龄、性别、肿瘤部位、病理分级和分期(UICC)等指标的相关性.结果:在41例胰腺癌组织中TGF-αTGF-β1的阳性表达率分别为73.2%、63.4%,在12例正常胰腺组织中的阳性表达率分别为16.7%、25.0%,两组之间存在显著差异(P=0.001).在胰腺癌组织中,TGF-β1的表达在不同分期中存在显著差异(P=0.019);TGF-α的表达与胰腺癌患者的年龄、性别、肿瘤部位、大小、分期及组织学分级无相关性(P>0.05).结论:TGF-α、TGF-β1在胰腺癌中高表达.TGF-β1与胰腺癌病理分期有关.     

固本颗粒胶囊对大鼠COPD模型血清IL-8及TNF-α影响的实验研究

目的:根据COPD慢性气道炎症学说,探讨固本颗粒胶囊对COPD模型大鼠血清中相关细胞因子的影响,从而阐明其防治COPD的作用机理,为固本颗粒胶囊的临床应用提供实验依据.方法:将50只Wistar大鼠随机分成正常对照组、模型对照组、强的松组、固本颗粒胶囊低剂量组、固本颗粒胶囊高剂量组,每组10只,以烟熏及气管内滴注脂多糖的方式建立大鼠COPD模型,予相应药物干预,观察记录大鼠一般状况,至实验结束,酶联免疫吸附法检测大鼠血清中相关细胞因子(IL-8、TNF-α)的含量.相关实验数据用SPSS10.0统计软件进行统计学处理.结果:大鼠一般状况及组织病理学改变提示COPD模型复制成功,与正常对照组比较,模型对照组大鼠血清中相关细胞因子(IL-8、TNF-α)的含量水平失衡.在改善COPD模型大鼠的一般状况、病理学改变及细胞因子含量比例失衡的作用结果来看,各治疗组与模型对照组均有显著差异(P＜0.05).综合比较,强的松组治疗作用最为显著,固本颗粒高剂量次之,固本颗粒低剂量组作用最弱.结论:①.固本颗粒胶囊能有效缓解COPD模型大鼠临床表现及气道炎症,其作用机制在于:能纠正COPD大鼠体内细胞因子水平失衡;②.上述作用,具有一定的量效关系,即随着给药剂量的增加有加强趋势.     

危重症患者肠功能障碍相关临床因素分析

目的:研究危重症患者肠功能障碍的相关临床因素,为有效防治肠功能障碍提供理论依据.方法:选取31例合并肠功能障碍的危重症患者为研究对象,以无肠功能障碍的30例危重症患者为对照.研究平均动脉压(MPA)、动脉血氧分压(PaO2),血白细胞计数、钾离子、GPT/GST、白蛋白等指标与肠功能衰竭发生的相关性;分析肠功能衰竭与脏器损伤个数及死亡率的关系.结果:伴有肠功能障碍的危重症患者MPA、PaO2和血清白蛋白水平低于对照组(P<0.05);血白细胞计数和GPT含量明显高于无肠功能障碍组(P<0.05);血钾离子浓度及GST两组无差异(P>0.05);发展至肠功能障碍的患者平均脏器损伤个数多于对照组,并且有着较高的死亡率(P<0.05).结论:伴有肠功能障碍的危重症患者早期就表现出严重的低血压,低血氧,低白蛋白血症,肝功能的损害,对于这些方面的监测既可以帮助我们早期发现肠功能障碍的存在,也有助于指导早期干预.     

肝细胞癌中NF-kBp65,VEGF和MMP-9蛋白的表达及其相关性研究

目的:探讨肝细胞癌(HCC)组织中NF-kBp65,VEGF和MMP-9的表达及其临床意义。方法:应用免疫组织化学(SP法)检测35例HCC组织,15例癌旁肝组织及10例正常肝组织中NF-kBp65,VEGF和MMP-9蛋白的表达并分析其与临床病理因素的关系。结果:NF-kBp65,VEGF和MMP-9蛋白在HCC组织中高表达率(分别为80%,74%,71%)高于对照组(P<0.01)。NF-kBp65的表达与HCC的Edmondson分级有关,即在Edmondson分级Ⅲ-Ⅳ的癌细胞中NF-kBp65的表达强于Edmondson分级Ⅰ-Ⅱ的(P<0.05);有静脉浸润的HCC组织中NF-kBp65,VEGF和MMP-9的表达高于无静脉浸润者,差异有统计学意义(P<0.05)。但NF-kBp65的表达与年龄,性别,肿瘤大小,AFP水平无关(P>0.05);VEGF和MMP-9的表达与年龄,性别,肿瘤大小,AFP水平,HCC的Edmondson分级无关(P>0.05)。NF-kBp65的表达与VEGF和MMP-9呈正相关(rs1=0.579,P1<0.05;rs2=0.406,P2<0.05)。结论:NF-kB...