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Expression of a functional recombinant vascular endothelial growth factor 165 (VEGF 165 ) in Arabidopsis thaliana 重组血管内皮生长因子165 (VEGF 165)在拟南芥中的表达
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2019-06-26 DOI: 10.1515/TJB-2017-0368
Qiang Weidong, F. Xue, Li Yixin, L. Xinxin, J. Kun, Sun Xiaoyu, C. Xi, Li Haiyan, Du Linna, Yang Jing
Objective: Targeting the protein of interest to a particular tissue to achieve high-level expression is an important strategy to increase expression efficiency. The use of the plant seed oil body as a bioreactor can not only increase the amount of target protein, but also reduce the cost of downstream processing. Methods: VEGF165 was expressed in Arabidopsis thaliana seeds via oilbody fusion technology. The pKO-VEGF165 vector was construted and transformed into A. thaliana seeds. T3 transgenic seeds was detected by SDS-PAGE and western blot methods. The cell activity was tested by MTT methods. Result: The phaseolin promoter was used to drive seedspecific expression of the VEGF165 gene in transgenic A. thaliana. The coding region of VEGF165 was fused to the Arabidopsis oleosin sequence to target the protein to the oil bodies in the seeds of transgenic plants. The T-DNA region of recombinant plasmid pKO-VEGF165 was shifted to A. thaliana seeds via the floral-dip method. Protein was analyzed by electrophoresis and protein hybridization analyses. Finally, MTT assays showed that the oleosinVEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro. Conclusion: Oleosin-VEGF165 was successfully expressed and it had stimulated HUVEC cell proliferation activity.
目的:将感兴趣的蛋白靶向特定组织实现高水平表达是提高表达效率的重要策略。利用植物籽油体作为生物反应器,不仅可以增加目标蛋白的量,还可以降低下游加工的成本。方法:采用油体融合技术在拟南芥种子中表达VEGF165。构建pKO-VEGF165载体,并将其转化为拟南芥种子。采用SDS-PAGE和western blot方法检测T3转基因种子。MTT法检测细胞活性。结果:利用phaseolin启动子驱动转基因拟南芥VEGF165基因的种子特异性表达。将VEGF165的编码区与拟南芥油酸蛋白序列融合,将该蛋白靶向转基因植物种子中的油酸体。通过花浸法将重组质粒pKO-VEGF165的T-DNA区转移到拟南芥种子中。蛋白电泳和蛋白杂交分析。最后,MTT实验表明,oleosinVEGF165融合蛋白在体外HUVEC细胞增殖中起作用。结论:Oleosin-VEGF165成功表达,并对HUVEC细胞增殖活性有促进作用。
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引用次数: 1
Toxicopathological evaluation of Picralima nitida seed aqueous extract in Wistar rats 小檗籽水提物对Wistar大鼠的毒性病理评价
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.83997
T. Sunmonu, O. B. Oloyede, T. Owolarafe, M. Yakubu, O. O. Dosumu
Objective: Picralima nitida is a widely used medicinal plant in West Africa for treating malaria, diarrhea and inflammation. The objective of this study is to evaluate the toxicological effect of aqueous seed extract of the plant in Wistar rats. Methods: Twenty-four apparently healthy animals were randomized into 4 groups comprising 6 rats each and orally administered with aqueous extract of P. nitida seeds at doses of 100, 200 and 400 mg/kg body weight with distilled water as control for 14 days. Specific liver and kidney function indices were assayed alongside haematological and histopathological analyses to monitor toxicity according to standard methods. Results: Phytochemical screening revealed the presence of alkaloids, glycosides, saponins, steroids and tannins. The extract had no significant effect on all kidney function indices assayed but caused a significant reduction (P < 0.05) in the activities of liver enzymes accompanied by significant decrease in liver to body weight ratio, serum total protein and globulin concentrations. No significant alteration was observed in the serum levels of albumin and conjugated bilirubin whereas the extract brought about significant increase (P < 0.05) in serum total bilirubin concentration. Haematological analysis revealed no significant effect on erythrocyte indices in contrast to white blood cell count and its differentials which were significantly elevated (P < 0.05) following extract administration. Histopathological studies further showed no distortion of cell structures in the studied organs. Conclusion: The available evidences in this study suggest that aqueous extract of P. nitida seeds exhibits mild and selective toxicity with liver as the target organ. Therefore, the herb may not be completely 'safe' as an oral remedy; and long term administration should be avoided.
目的:黑皮草是西非地区广泛使用的一种治疗疟疾、腹泻和炎症的药用植物。本研究的目的是评价该植物种子水提物对Wistar大鼠的毒理学作用。方法:将24只健康动物随机分为4组,每组6只,分别以100、200、400 mg/kg体重的剂量口服牛油果种子水提物,以蒸馏水为对照,连续14 d。按标准方法测定肝肾功能指标及血液学和组织病理学指标,监测毒性。结果:通过植物化学筛选,发现其含有生物碱、糖苷、皂苷、甾体和单宁。各肾脏功能指标均无显著变化,但肝酶活性显著降低(P < 0.05),肝重比、血清总蛋白和球蛋白浓度显著降低。血清白蛋白和结合胆红素水平无显著变化,而血清总胆红素浓度显著升高(P < 0.05)。血液学分析显示,提取物对红细胞指数无显著影响,而白细胞计数及其差异显著升高(P < 0.05)。组织病理学研究进一步表明,所研究器官的细胞结构未发生畸变。结论:本研究证据表明,硝酸菌种子水提物具有以肝脏为靶器官的轻度选择性毒性。因此,作为口服药物,这种草药可能并不完全“安全”;应避免长期用药。
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引用次数: 7
Effects of Dietary Chicken Grill Oil and Sunflower Seed Oil on Blood and Liver Oxidant/Antioxidant Status and Liver Function Tests in Laying Japanese Quails (Coturnix coturnix japonica) 鸡扒油和葵花籽油对产蛋日本鹌鹑血液和肝脏氧化/抗氧化状态及肝功能的影响
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.81894
T. Buyukoglu
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引用次数: 1
Biosorption Studies For Removal of Cu (II) ions Onto Saccharum Bengalense an Efficient and Eco-Friendly Biosorbent 高效环保生物吸附剂班加兰糖对Cu (II)离子的生物吸附研究
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.25483
M. I. Din
Objective: In this research work, a low-cost biosorbent derived from the pulp of Saccharum bengalense (SB) was used as an adsorbent material for the removal of Cu (II) ions from aqueous solution. Methods: A batch adsorption method was experimented for biosorptive removal of copper ions from the aqueous solution. Results: Nearly 90% removal of copper is possible at initial pH 6.0, under the batch test conditions. Langmuir, Freundlich and Dubinin–Radushkevich (D–R) adsorption isotherms have been applied to describe the biosorption of Cu (II) by SB. It was found that biosorption of Cu (II) by SB followed Langmuir and Freundlich model. The monolayer adsorption capacity of SB as obtained from Langmuir isotherm at 40 0C was found to be 10.42 mg/g at 323K. The Cu (II) adsorption data was analyzed by applying pseudo-first-order and pseudo-second-order kinetic models. The intraparticle diffusion model was applied to investigate the rate-determining step. It was found that the biosorption of Cu (II) ions on SB followed pseudo second-order rate kinetics. Thermodynamic parameters such as free energy change (ΔG°), enthalpy change (ΔH°) and entropy change (ΔS°) have been calculated respectively revealed the spontaneous, feasible and endothermic nature of adsorption process. Conclusion: The results of the present investigation suggested that biosorbent SB can be used as an economically feasible and environmentally friendly biosorbent for the removal of Cu (II) from aqueous solutions.
目的:研究以蔗糖(Saccharum bengalense,简称SB)为原料制备的低成本生物吸附剂,用于去除水中的Cu (II)离子。方法:采用间歇式吸附法对铜离子进行生物吸附去除。结果:在初始pH 6.0条件下,在批量试验条件下,铜的去除率接近90%。采用Langmuir, Freundlich和Dubinin-Radushkevich (D-R)等温线描述了SB对Cu (II)的生物吸附,发现SB对Cu (II)的生物吸附符合Langmuir和Freundlich模型。通过Langmuir等温线在40℃时对SB的单层吸附量在323K时为10.42 mg/g。采用拟一级和拟二级动力学模型对Cu (II)吸附数据进行了分析。采用颗粒内扩散模型对速率决定步骤进行了研究。发现SB对Cu (II)离子的吸附符合准二级速率动力学。分别计算了吸附过程的自由能变化(ΔG°)、焓变化(ΔH°)和熵变化(ΔS°)等热力学参数,揭示了吸附过程的自发性、可行性和吸热性。结论:生物吸附剂SB是一种经济、环保的脱除水中Cu (II)的生物吸附剂。
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引用次数: 2
PKC stimulation increases expression of cholinesterases in R28 cell line PKC刺激增加R28细胞系胆碱酯酶的表达
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.38981
E. Bodur, P. Layer
Objective: Previously we have shown a counter-regulation between cholinesterases (ChEs) through both anti-sense Butyrylcholinesterase (BChE) transfection and knock-down by siR- NAs in R28 cells. In course of this counter-regulation, the status of the cell growth- and dif- ferentiation-related signaling pathways PKC and ERK were also changed. Down-regulation of BChE led to an increased PKC-α expression. Here, we demonstrate that this interaction between ChEs and PKC is reciprocal. Methods: R28 cells were treated with final 10 µM Di-octanoyl glycerol (DOG) and 10 nM siRNAs against BChE. Expression analysis was done by RT-PCR, Western Blot, IHC and activity assays. Results: DOG treatment along with BChE knockdown resulted in increased PKC-α expres- sion, as compared with DOG treatment alone. Change in ERK1 expression was not as pro- found. In R28 cells, DOG stimulation led to marked rapid increase in both AChE and BChE expression.
目的:先前我们通过反义丁基胆碱酯酶(BChE)转染和siR- NAs敲除R28细胞中胆碱酯酶(ChEs)之间的反向调控。在这种反调控过程中,细胞生长和分化相关的信号通路PKC和ERK的状态也发生了变化。下调BChE导致PKC-α表达增加。在这里,我们证明了che和PKC之间的相互作用是相互的。方法:用10µM二辛烷酰甘油(DOG)和10 nM sirna对R28细胞进行抗BChE作用。采用RT-PCR、Western Blot、免疫组化和活性检测进行表达分析。结果:与单独治疗相比,DOG联合BChE敲低可导致PKC-α表达增加。ERK1的表达变化不明显。在R28细胞中,DOG刺激导致AChE和BChE表达显著快速增加。
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引用次数: 1
The effect of L-carnitine on nitric oxide metabolism in streptozotocin-induced diabetic rats 左旋肉碱对链脲佐菌素诱导的糖尿病大鼠一氧化氮代谢的影响
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.04934
G. S. Ozgun
Objective: The aim of this study is to investigate the effect of L-carnitine on plasma and liver nitric oxide metabolism in streptozotocin-induced diabetic rats. Methods: Sprague Dawley female rats were divided randomly into following groups: control, Lcarnitine, diabetes and diabetes+L-carnitine. Diabetes and diabetes+L-carnitine groups were intraperitonally injected with a single dose of streptozotocin (40 mg/kg) prepared in the citrate buffer (pH 4.5). Other groups were injected with only citrate buffer. 72 hours after the streptozotocin injection, Lcarnitine (500 mg/kg/day) was given intraperitoneally to L-carnitine and diabetes+L-carnitine groups for 15 days. Physiological saline was given intraperitoneally to the other groups for 15 days. Blood sugar (at 72 hours and the end of experiment), liver nitric oxide and inducible nitric oxide synthase, plasma nitric oxide and nitrotyrosine levels were measured. Results: Blood glucose levels in diabetic groups were higher compared with other groups. Percentage change of blood glucose in diabetes+L-carnitine group was lower compared with other groups. Also diabetes+L-carnitine group’s plasma nitric oxide levels were higher than control group. Plasma nitrotyrosine levels of L-carnitine injected groups were lower than diabetes group. There was no significant difference between the levels of liver inducible nitric oxide synthase and nitric oxide in groups. Conclusion: As a result, our study showed that plasma and liver nitric oxide and liver inducible nitric oxide synthase levels aren’t changed significantly but plasma nitrotyrosine levels are increased at the end of 15th day of experimental diabetes. On the other hand, our results also showed that L-carnitine causes an increase in plasma nitric oxide levels and a decrease in plasma nitrotyrosine levels whereas it has no effect on liver nitric oxide and inducible nitric oxide synthase levels.
目的:探讨左旋肉碱对链脲佐菌素诱导的糖尿病大鼠血浆和肝脏一氧化氮代谢的影响。方法:将雌性sd大鼠随机分为对照组、左旋肉碱组、糖尿病组和糖尿病+左旋肉碱组。糖尿病组和糖尿病+左旋肉碱组腹腔注射单剂量链脲佐菌素(40 mg/kg),用柠檬酸缓冲液(pH 4.5)配制。其他组只注射柠檬酸缓冲液。注射链脲佐菌素72 h后,左旋肉碱组和糖尿病+左旋肉碱组腹腔注射左旋肉碱500 mg/kg/d,连续15 d。其余各组腹腔灌胃生理盐水15 d。测定大鼠72h及实验结束时的血糖、肝脏一氧化氮和诱导型一氧化氮合酶、血浆一氧化氮和硝基酪氨酸水平。结果:糖尿病组血糖水平高于其他各组。糖尿病+左旋肉碱组血糖变化百分比较其他组低。糖尿病+左旋肉碱组血浆一氧化氮水平高于对照组。左旋肉碱注射组血浆硝基酪氨酸水平低于糖尿病组。肝诱导型一氧化氮合酶和一氧化氮水平在各组间无显著差异。结论:我们的研究表明,在实验糖尿病第15天结束时,血浆和肝脏一氧化氮和肝诱导型一氧化氮合酶水平没有显著变化,但血浆硝基酪氨酸水平升高。另一方面,我们的研究结果还表明,左旋肉碱引起血浆一氧化氮水平升高和血浆硝基酪氨酸水平降低,而对肝脏一氧化氮和诱导型一氧化氮合酶水平没有影响。
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引用次数: 2
Platelet function tests 血小板功能试验
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2015.17894
Z. Dikmen
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引用次数: 0
Real-Time PCR Analysis of Pyrethroid Resistance in Helicoverpa armigera from Turkey 土耳其棉铃虫拟除虫菊酯抗性的实时PCR分析
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.50490
M. Konuş, S. U. Karaağaç, M. Işcan
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引用次数: 1
Platelet function disorders 血小板功能障碍
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/tjb.2015.80958
Z. Dikmen
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引用次数: 0
Genotype and allele frequencies of the intestinal fatty acid binding protein-2 gene in two Arab populations 两个阿拉伯人群肠道脂肪酸结合蛋白-2基因的基因型和等位基因频率
IF 0.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2014-01-01 DOI: 10.5505/TJB.2014.15870
A. Salem
Objective: Intestinal fatty acid binding protein (IFAPB) participates in the uptake, intracellular metabolism and/or transport of long chain fatty acids. A polymorphism at codon 54 in exon 2 of FABP2 gene, which encodes for the IFAPB, exchanges an Alanine for Threonine. FABP2 gene polymorphism could modify the uptake of fatty acids, and it could correlate with risk of several diseases. In the light of the potential role of the FABP2 polymorphism, the aim of this study was to determine the frequency of the Ala54Thr FABP2 polymorphism in two Middle Eastern Arab Populations. Materials and Methods: Genotyping was investigated in 182 and 120 unrelated healthy subjects from Bahrain and Jordan, respectively. A PCR-RFLP assay was applied for determination of Ala54Thr (rs1799883) FABP2 polymorphism. Allele frequencies were calculated by direct counting. Hardy Weinberg Equilibrium was evaluated using a Chi- square goodness of fit test. Results: In the studied Bahraini subjects, 52.8% were homozygous for the Ala54/Ala54 genotype, 35.7% were heterozygous for the Ala54/Thr54 genotype and 11.5% were homozygous for the Thr54/Thr54 genotype. The gene frequencies obtained in Jordanians were: 48.3%, 43.3% and 8.4% for Ala54/Ala54; Ala54/Thr54 and Thr54/Thr54 genotypes, respectively. The frequencies of the allele Ala54 and the allele Thr54 of the FABP2 gene were found to be 0.706 and 0.294 for Bahrainis and 0.700 and 0.300 for Jordanians. These results revealed a similar population polymorphism frequency as in previous European and Arab populations' studies. Conclusion: This is the first study to investigate the population frequency of the Thr54 allele in Bahraini and Jordanian populations.
目的:肠道脂肪酸结合蛋白(IFAPB)参与长链脂肪酸的摄取、胞内代谢和/或转运。编码IFAPB的FABP2基因2外显子密码子54的多态性将丙氨酸交换为苏氨酸。FABP2基因多态性可以改变脂肪酸的摄取,并可能与多种疾病的风险相关。鉴于FABP2多态性的潜在作用,本研究的目的是确定两个中东阿拉伯人群中Ala54Thr FABP2多态性的频率。材料与方法:分别对来自巴林和约旦的182名和120名无血缘关系的健康受试者进行基因分型研究。采用PCR-RFLP法检测Ala54Thr (rs1799883) FABP2多态性。采用直接计数法计算等位基因频率。采用卡方拟合优度检验评价哈代温伯格均衡。结果:巴林人Ala54/Ala54基因型纯合子占52.8%,Ala54/Thr54基因型杂合子占35.7%,Thr54/Thr54基因型纯合子占11.5%。约旦人Ala54/Ala54基因频率分别为48.3%、43.3%和8.4%;分别为Ala54/Thr54和Thr54/Thr54基因型。发现FABP2基因的等位基因Ala54和Thr54的频率在巴林人中分别为0.706和0.294,在约旦人中分别为0.700和0.300。这些结果揭示了与先前欧洲和阿拉伯人群研究相似的群体多态性频率。结论:本研究首次调查了巴林和约旦人群中Thr54等位基因的群体频率。
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引用次数: 0
期刊
Turkish Journal of Biochemistry-turk Biyokimya Dergisi
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