人胎盘TRAIL基因的cDNA克隆及序列测定     

张明烽  唐爽  宋红卫 《西北农林科技大学学报(自然科学版)》2005,33(2):6-8,13

根据GeneBank(U37518)中TRAIL(肿瘤坏死因子相关的凋亡诱导配体)的cDNA序列,设计扩增引物。采用RT-PCR从人胎盘中扩增出TRAIL基因的全长cDNA。产物纯化后连至pGEM-TEasy载体,转化大肠杆菌DH5α。筛选阳性克隆菌,酶切鉴定并进行序列测定。结果表明,本试验成功地克隆了TRAIL基因的1039bp全长cDNA。  相似文献   

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells     

Fei Li  Yanan Yang  Panpan Zhu  Weina Chen  Dongli Qi  Xiupu Shi  Chunfeng Zhang  Zhonglin Yang  Ping Li 《Fitoterapia》2012

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.  相似文献   

OPG/RANKL/RANK系统的生理功能          下载免费PDF全文

王卫中 《家畜生态学报》2012,33(4):102-105,120

骨是一个代谢活跃的器官,在整个生命周期中都在进行不断的更新与重建。在骨形态发生和重建的过程中主要受到细胞因子和激素的调节,同时骨代谢也受到OPG/RANKL/RANK系统的调控。论文主要对骨骼生理结构与骨组织细胞之间的关系,骨代谢和骨的形成过程,OPG/RANKL/RANK系统的生理功能,OPG/RANKL/RANK系统的影响因子以及骨代谢紊乱所引起的疾病几个方面作一综述。  相似文献   

Evidence of dissolved trivalent manganese in acidic forest soils     

Constantin Lux  Tim Mansfeldt 《植物养料与土壤学杂志》2023,186(3):321-329

Background

Evidence of trivalent manganese (Mn³⁺) in the aqueous phase of soils is unknown so far although this strong oxidant has large environmental implications.

Aims

We aimed to modify a spectrophotometric protocol (porphyrin method) and to discriminate between Mn²⁺ and Mn³⁺ in the aqueous phase of forest soils based on kinetic modeling.

Methods

We investigated manganese speciation in 12 forest floor solutions and 41 soil solutions from an acidic forest site by adjusting pH and correcting for absorbance.

Results

The solutions showed broad ranges in pH (3.4−6.3), dissolved organic carbon (DOC, 1.78−77.1 mg C L⁻¹), and total Mn (Mn_T, 23.9−908 µg L⁻¹). For acidic solutions, a pH-buffer was added to increase the pH of the solutions to 7.5−8.0, and background absorption was corrected for colored solutions, that is, solutions high in DOC. This was done to accelerate the reaction kinetics and avoid overestimation of Mn_T concentrations. After the pH and color adjustments, the comparison of Mn_T concentrations between the porphyrin method and optical emission spectrometry showed good agreement. Trivalent Mn, which is stabilized by organic ligands, constitutes significant proportions in both forest floor solutions (10−87%) and soil solutions (0.5−74%).

Conclusions

The dissolved Mn³⁺ is present in acidic forest soils. Thus, we revise the paradigm that this species is not stable and encourage to apply the revised method to other soils.  相似文献   

Kinetics of Ligand-Controlled Release of Zinc in Acid Sulfate Paddy Soils     

Worachart WISAWAPIPAT  Aksarapak PONGPOM 《土壤圈》2019,29(2):216-223

The elevated solubility of zinc(Zn) in acid sulfate paddy soils can limit rice production and pose a risk of environmental pollution.However, little attention has been paid to the ligand-controlled release of Zn in these soils. Here we quantified the rate of ligandcontrolled Zn release in Thai acid sulfate paddy soils, using ethylenediaminetetraacetic acid as the extractant. Sequential extractions were performed to obtain quantitative information on Zn fractions contributing to the ligand-controlled mechanisms. The amount of Zn released within 192 h varied significantly(8–43 mg kg~(-1)), which corresponded to 12%–40% of the total soil Zn, indicating that Zn solubility in most soils was relatively low and that Zn mainly occurred as residual phases. The kinetics of Zn release was well described by the power function model(r = 0.65–0.99, median = 0.87). The magnitude of initial Zn release(coefficient a) was significantly(P 0.05) related to the aqua regia-soluble Zn. Easily mobile Zn, organically bound Zn, and Zn associated with Fe and Mn oxides also contributed to the ligand-controlled release mechanisms to various degrees. Our results provide a systematic understanding of Zn fractions and release from acid sulfate paddy soils, the dynamics of which have a significant influence on the availability, phytoextraction, and mobility of Zn in terrestrial and engineered environments.  相似文献   

捻转血矛线虫Hc-FAR-4蛋白的表达特性及其与配体结合能力分析     

韦海典  陈学秋  黄艳  时恒枝  周静茹  吴飞  杜爱芳  杨怡 《中国农业科学》2019,52(17):3059-3068

【目的】捻转血矛线虫(Haemonchus contortus)寄生在牛羊等反刍动物的皱胃中,引起的捻转血矛线虫病在我国呈全国性流行。文章通过研究脂肪酸与视黄醇结合相关蛋白Hc-FAR-4的表达特性及配体结合能力,以了解其在捻转血矛线虫生长、发育、繁殖过程中的作用。【方法】对Hc-far-4进行克隆、并且构建原核表达载体pET-30a-Hc-far-4,pET-30a-Hc-far-4重组质粒经过PCR与双酶切鉴定准确无误之后转化到E. coli BL21感受态细胞中,用0.1 mmol·L ^-1IPTG(isopropyl-β-d-thiogalactoside)进行重组蛋白的诱导表达。收集重组蛋白rHc-FAR-4,利用荧光分析法,研究rHc-FAR-4蛋白与DAUDA、视黄醇、油酸的结合能力。配体结合试验是依据荧光物质retinol与脂肪酸类似物DAUDA在极性和非极性溶液中,在某一波长的激发光激发下,所发出的荧光光谱随之变化的特性,判断rHc-FAR-4蛋白是否具有与DAUDA、retinol结合的能力。再在体系中加入非荧光长链脂肪酸油酸,根据荧光光谱的变化情况判定油酸能否分别与DAUDA、retinol竞争目的蛋白的脂肪酸结合位点,间接说明目的蛋白能否与非荧光脂肪酸油酸结合。同时制备鼠源多克隆抗体,利用ELISA技术检验免疫后小鼠的抗体效价,抗体效价合适即可收集小鼠血清。收集的血清用于免疫组织荧光（IHF）试验,探究Hc-FAR-4的表达部位,从而推测其功能。该试验过程为：对捻转血矛线虫进行石蜡包埋,石蜡切片,抗原修复,3%的BSA 于4℃封闭过夜,自制鼠源多克隆抗体作为一抗孵育1 h; Alexa Fluor? 488 nm羊抗鼠IgG作为二抗避光孵育1 h,DAPI染色30 min,激光共聚焦显微镜检查染色情况。利用荧光定量PCR技术分析Hc-far-4在捻转血矛线虫各个主要阶段的表达特性。【结果】成功克隆目的基因Hc-far-4,测序结果与Sanger数据库中捻转血矛线虫far-4基因序列（>HCISE00908800.t1）相似度为 99.9%;重组质粒 pET-30a-Hc-far-4在E. coli BL21中成功表达,在诱导8 h后达到峰值。经过ELISA检测,结果显示制备的鼠源多克隆抗体效价达到1﹕1 024 000—1﹕2 048 000,可用于后续试验。Western Blot鉴定结果显示rHc-FAR-4重组蛋白带有His标签,并且条带大小为25 kD,结果符合预期。制备的鼠源多克隆抗体经过Western Blot鉴定,能够与天然Hc-FAR-4 蛋白结合,说明该抗体可用于IHF试验。配体结合试验结果表明 rHc-FAR-4蛋白具有结合脂肪酸与视黄醇的能力。荧光定量PCR分析表明,Hc-far-4在四期幼虫中的转录水平最高;IHF试验表明,Hc-FAR-4主要表达在捻转血矛线虫的肠壁、性腺中。综上推测Hc-FAR-4蛋白主要在寄生生活阶段参与了转运脂肪酸与视黄醇,为捻转血矛线虫的生长发育与生殖提供营养物质。【结论】捻转血矛线虫 FAR-4 蛋白能够结合脂肪酸类似物DAUDA和视黄醇,但是与油酸的结合能力较弱,主要表达在肠壁,在性腺、角皮中也有少量表达,其基因在进行营寄生生活阶段时表达量达到峰值。  相似文献   

The effect of TNF-related apoptosis-inducing ligand on the activity of nuclear kappa B in prostate cancer cell line PC-3M     

ZHONG Wei  HE Wei  MA Wang  ZHANG Ke-liang  WEI Shao-zhong  CHEN Zhao-hui  CHEN Xiao-chun  ZENG Fu-qing  XIAO Chuan-guo△ 《园艺学报》2008,24(5):966-971

AIM: To investigate the activation and inactivation of nuclear factor kappa B (NF-κB) when tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is applied to induce the apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: After the treatment of TRAIL or LPS at different doses, we tested the nuclear translocation of NF-κB by cell immunohistochemical staining and electrophoretic mobility shift assay(EMSA), and evaluated the level of IκB by RT-PCR under pyrrolidine dithiocarbamate (PDTC) treatment. RESULTS: EMSA and cell immunohistochemical analysis showed that the translocation of NF-κB was significantly activated when PC-3M cells were treated with TRAIL or LPS (P<0.05). The pretreatment of PDTC upregulated the expression of IκB and blocked the nuclear translocation of NF-κB.CONCLUSION: TRAIL remarkably stimulates the activation of nuclear NF-κB in androgen-independent prostate cancer cells. On the other hand, the translocation of NF-κB can be significantly and efficiently inhibited in PC-3M cells by pretreatment with PDTC. The increased expression of IκB might be a clue for this inhibition, which means the possible way to enhance the effect of TRAIL in the apoptosis of prostate cancer cells.  相似文献   

Polymorphism in exon 2 encoding the putative ligand binding pocket of the bovine insulin‐like growth factor 1 receptor affects milk traits in four different cattle breeds          下载免费PDF全文

M. Szewczuk 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2017,134(1):34-42

  相似文献   

Wastewater Chemistry and Fractionation of Bioactive Phosphorus in Dairy Manure     

《Communications in Soil Science and Plant Analysis》2012,43(7-8):907-924

Abstract

Because manure organic phosphorus (P) is environmentally bioactive, a novel in situ enzyme hydrolysis assay was developed to identify water soluble‐ and labile complexed P and mechanisms controlling P solubilization in dairy manure. Water‐extractable P averaged 16% (±14.8%) of total P of 107 manures collected across five northeastern states of the USA. Adding a multidentate ligand solubilized inorganic complexed P (15±8.3%) primarily associated with calcium (Ca) and magnesium (Mg). An additional organic fraction (35.9±15.6%) was hydrolyzable by fungal phytases. The assay was more revealing about on‐farm P management than just knowing total P; the water‐extractable P distribution was skewed to the left, and two thirds were <2500 mg kg^?1; bioactive and total P were normally distributed, differing extensively between farms with a range spanning an order of magnitude. The assay's simplicity and robustness over the wide range of manure characteristics may increase routine evaluation of whole‐farm accumulation of environmentally sensitive P forms.  相似文献   

低分子量有机配体对黏粒矿物吸附苏云金芽孢杆菌的影响   总被引：2，自引：0，他引：2  

吴华勇  蒋代华  蔡鹏  荣兴民  黄巧云 《土壤学报》2011,48(6):1293-1297

土壤是微生物的理想栖息地,土壤微生物群落组成复杂,数量巨大。1g土壤中可能栖息着上百亿个微生物,其中细菌可达1010个[1]。约80%～90%的土壤微生物寄居于土壤固相表面,如黏土矿物、金属氧化物或有机质表面[2]。细菌与矿物间的相互作用在土壤污染物转化与降解、团聚体形成[3]、矿物风化[4-6]及病原菌运移[7-8]等诸多过程起着关键作用。  相似文献