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Study on the Simultaneously Quantitative Detection for β-Lactoglobulin and Lactoferrin of Cow Milk by Using Protein Chip Technique 被引量:3
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作者 YIN Ji Yong HUO Jun Sheng +2 位作者 MA Xin Xin SUN Jing HUANG Jian 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2017年第12期875-886,共12页
Objective To research a protein chip method which can simultaneously quantitative detect β-Lactoglobulin (β-L) and Lactoferrin (Lf) at one time. Methods Protein chip printer was used to print both anti-β-L anti... Objective To research a protein chip method which can simultaneously quantitative detect β-Lactoglobulin (β-L) and Lactoferrin (Lf) at one time. Methods Protein chip printer was used to print both anti-β-L antibodies and anti-Lf antibodies on each block of protein chip. And then an improved sandwich detection method was applied while the other two detecting antibodies for the two antigens were added in the block after they were mixed. The detection conditions of the quantitative detection for simultaneous measurement of β-L and Lf with protein chip were optimized and evaluated. Based on these detected conditions, two standard curves of the two proteins were simultaneously established on one protein chip. Finally, the new detection method was evaluated by using the analysis of precision and accuracy. Results By comparison experiment, mouse monoclonal antibodies of the two antigens were chosen as the printing probe. The concentrations of β-L and Lf probes were 0.5 mg/mL and 0.5 mg/mL, respectively, while the titers of detection antibodies both of β-L and Lf were 1:2,000. Intra- and inter-assay variability was between 4.88% and 38.33% for all tests. The regression coefficients of protein chip comparing with ELISA for β-L and Lf were better than 0.734, and both of the two regression coefficients were statistically significant (r = 0.734, t = 2.644, P = 0.038; and r = 0.774, t = 2.998, P = 0.024). Conclusion A protein chip method of simultaneously quantitative detection for β-L and Lf has been established and this method is worthy in further application. 展开更多
关键词 protein chip Simultaneously Quantitative detection Β-LACTOGLOBULIN LACTOFERRIN
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Weak cation exchange 2 protein chip for detecting differentially expressed brainstem proteins in a rat model of closed traumatic brain injury
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作者 Lin Liang Haiying Gong +2 位作者 Li Zhan Shuwang Yang Yongliang Zhang 《Neural Regeneration Research》 SCIE CAS CSCD 2010年第5期372-377,共6页
BACKGROUND: Studies have reported the combined use of two-dimensional gel electrophoresis and mass spectrometry to detect differentially expressed proteins in the rat brainstem following brain injury. However, the de... BACKGROUND: Studies have reported the combined use of two-dimensional gel electrophoresis and mass spectrometry to detect differentially expressed proteins in the rat brainstem following brain injury. However, the detected differential proteins often exhibit low sensitivity and high relative molecular weight. Although protein chip technology is thought to compensate for these inadequacies, no related studies or results have been reported. OBJECTIVE: To propose the application of weak cation exchange protein chips in combination with mass spectrometry for determining protein expression profiles and characteristics in the brainstem following closed brain injury. DESIGN, TIME AND SETTING: Randomized, controlled, animal experiments utilizing proteomics were performed from June 2007 to December 2008 in the Proteomics Laboratory, Medical College of Chinese People's Armed Police Force. MATERIALS: Weak cation exchange 2 protein chip, Ciphergen Proteinchip System (PBS-IIC). METHODS: A total of 72 rats were randomly assigned to two groups: sham-surgery (n = 12) and injury (n = 60). A closed traumatic brain injury model caused by falling object was replicated in the injury group, which was then subdivided into five subgroups according to different time points after injury: 4, 8, 12, 24, and 48 hours, with 12 rats in each subgroup. In the sham-surgery group, only the skin was removed and the stainless steel pad was fixed to the skull. MAIN OUTCOME MEASURES: The brain injury rats were sacrificed at 4, 8, 12, 24, and 48 hours after injury, respectively, and the control rats were sacrificed at 24 hours. Pathological changes in the brainstem were determined using hematoxylin-eosin staining, and differential protein expression in the brainstem was detected using a weak cation exchange 2 protein chip and protein chip reader. RESULTS: In the sham-surgery group, cells appeared normal. However, in the brain injury group, some brainstem neurons exhibited pyknosis, with reduced numbers of Nissl bodies in the cytoplasm swollen cell bodies and nuclei, irregular staining in the cytoplasm, and decreased numbers of neurons. Results from weak cation exchange 2 protein chip detection demonstrated that, compared with the sham-surgery group, the expression profiles of 2 proteins were altered in the brainstem of the injury group. At 12, 24, and 48 hours after injury, expression was increased (P 〈 0.01 ). The mass charge ratio (M/Z) of 7 862 differentially expressed proteins was greater in the sham-surgery group compared with 12 and 24 hours after injury (P 〈 0.05). CONCLUSION: The combined method of weak cation exchange 2 protein chip and mass spectrometry detected differential protein expression in the brainstem following closed brain injury in the rats, which suggested that closed brain injury induced altered protein expression profiles in the brainstem. 展开更多
关键词 protein chip ion exchange brain injury BRAINSTEM rats PROTEOMICS neural regeneration
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Proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus,as demonstrated by the surface enhanced laser desorption/ionization(SELDI)protein chip system
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作者 ZHI JUN LIU BIN WANG +5 位作者 YING TIAN ZHI QIANG BAI SHOU YI DING Xu XIA SONG ZHI YONG YAN DONG MENG QIAN 《Journal of Microbiology and Immunology》 2007年第1期46-51,共6页
The proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus (HCMV) was investigated at the protein level by using the surface enhanced laser desorp... The proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus (HCMV) was investigated at the protein level by using the surface enhanced laser desorption/ionization (SELDI) protein chip system in order to develop a method of study for the pathogenesis of HCMV infection. In this study, the cultured U251 cells were infected with HCMV in good condition and the supernatants of lysates and the extracellular fluids of the cultivated infected cells were quantitatively defined for the expressed proteins. The proteomics of the differential protein expression in cells before and after infection was analyzed by WCX2 arrays on the protein chip reader. It was demonstrated that the eytopathic effects of infected cells appeared on the 5th day after infection, however, the differential protein expression was evident at 6 h after infection as revealed by RT-PCR and mass spectrometry. The protein peaks captured from different batches of samples, from the same sample detected with different arrays or for the different times were all equivalent. With the molecular weight range from 2000 Da to 3000 Da, chip captured 82 peaks from the intracellular fluids and 11 protein peak from the cellular fluid in which compared with the control group, the protein peaks with molecular weight of 13 536.3 Da, 10 046.1 Da and 17 106.2 Da were close to those of β-amyloid protein, caspase-1 precursor and LPS-induced TNF-α factor respectively, which showed brief up-regulation 4 h after infection, and continued to raise 48 h later. These results infer that these proteins may be related to the apoptosis induced by HCMV infection, thus suggesting that the apoptosis induced by HCMV infection may play a role in the pathogenesis of HCMV infection. 展开更多
关键词 SELDI protein chip U251 cells HCMV protein expression
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替普瑞酮通过E3泛素连接酶CHIP减轻LPS引起的心肌炎症反应和心功能障碍
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作者 徐丽婷 刘颖文 +8 位作者 李健玲 林婉 王淼 余蕾 张雪 李航 王华东 吕秀秀 王一阳 《中国病理生理杂志》 CAS CSCD 北大核心 2024年第5期862-871,共10页
目的:探究替普瑞酮又称香叶基香叶基丙酮,GGA)对脂多糖(LPS)诱导的心功能障碍的治疗作用及机制。方法:(1)取8周龄C57BL/6雄性野生型小鼠和热休克蛋白70(HSP70)羧基末端相互作用蛋白(CHIP)基因敲除小鼠,随机分为对照组、LPS组、LPS+GGA组... 目的:探究替普瑞酮又称香叶基香叶基丙酮,GGA)对脂多糖(LPS)诱导的心功能障碍的治疗作用及机制。方法:(1)取8周龄C57BL/6雄性野生型小鼠和热休克蛋白70(HSP70)羧基末端相互作用蛋白(CHIP)基因敲除小鼠,随机分为对照组、LPS组、LPS+GGA组和GGA组,每组8只。用腹腔注射LPS(25 mg/kg)的方法建立模型,于LPS刺激后1 h给予小鼠腹腔注射GGA(100 mg/kg)。利用小动物超声系统评估小鼠心脏功能;采集各组小鼠血清,检测血清肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)水平;HE染色观察病理学改变;ELISA检测心脏组织中炎症因子肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的水平;Western blot检测各组心脏组织HSP70、CHIP、核转运蛋白α2(KPNA2)、髓过氧化物酶(MPO)、血管细胞黏附分子(VCAM)和细胞间黏附分子(ICAM)的蛋白表达以及细胞核NF-κB的水平。(2)利用小鼠心肌细胞HL-1,建立LPS刺激的离体细胞炎症模型。ELISA检测细胞上清中TNF-α和IL-6的水平;Western blot检测心肌细胞中HSP70、CHIP和KPNA2蛋白表达;免疫荧光染色观察细胞核NF-κB的表达。结果:(1)GGA有效改善LPS刺激小鼠的心脏功能,显著提高射血分数和左室短轴缩短率(P<0.01),减少血清CK-MB和LDH含量(P<0.01),减轻心肌损伤。(2)GGA显著减少LPS引起的TNF-α和IL-6炎症因子的释放(P<0.01),以及NF-κB的入核,降低心肌组织中KPNA2、MPO、VCAM和ICAM蛋白表达,增加心肌组织和细胞HSP70的水平(P<0.01)。(3)在CHIP基因敲除的心肌细胞和小鼠中,GGA不能抑制LPS引起的炎症反应,失去了改善LPS刺激小鼠心脏功能的作用。结论:GGA能够减轻LPS引起的心功能障碍,其作用机制与升高HSP70的表达,促进CHIP的活化,减少NF-κB的入核,抑制炎症因子的释放有关。CHIP的敲除使GGA丧失了减轻LPS诱导的炎症反应和心肌损伤的作用。 展开更多
关键词 替普瑞酮 脂多糖 心功能障碍 chip蛋白 炎症
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Measures of bioavailable serum sex hormone levels in aging Chinese by protein chip
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作者 ZHOU Yong, CHANG Shuying, MENG Xiaoluo, YU Huafeng, WANG Luning, HE Jinggui, ZHANG Baohe, ZHANG Juntian, GENG Meiyu & DU Guanhua Institute of Marine Drugs and Food, Ocean University of China, Qingdao 266003, China Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China +3 位作者 General Hospital of Air Force, PLA, Beijing 100036, China 304 Hospital of PLA, Beijing 100037, China Beijing Tongren Hospital, Beijing 100730, China General Hospital of PLA, Beijing 100853, China 《Science China(Life Sciences)》 SCIE CAS 2006年第3期286-292,共7页
The purpose of this study was to develop a protein chip technique based on receptor binding assays to measure bioavailable serum sex hormone levels (BSSHL). 224 aging healthy Chinese were investigated to get the refer... The purpose of this study was to develop a protein chip technique based on receptor binding assays to measure bioavailable serum sex hormone levels (BSSHL). 224 aging healthy Chinese were investigated to get the referenced values of BSSHL for the first time. In the assays recombined sex hormone receptor proteins were jointed to polysaccharide coated slides to make protein chip,; the dose-dependence curves of sex hormone on chip were prepared. The data showed that this method had good precision (CV>16%); accuracy (Bias>10%),; the sensitivity could reach 1 pmol/L. From the results, BSSHL of men; women declined with aging, but no significant differences were observed. The BSSHL of aging men were higher than those of women. The bioavailable serum;rogen level of men was 52–112 pmol/L, women’s was 3–70 pmol/L; the whole group was 41.9–81.4 pmol/L. The bioavailable serum estrogen level of men was 0.8–3.0 pmol/L, women’s was 1.2–2.5 pmol/L; the whole group was 0.6–2.64 pmol/L. Based on the assays, BSSHL measurement by protein chip can meet the needs of epidemiological studies in terms of speed, accuracy; sample volume required,; was helpful in quantitative assessment of aging people’s health. 展开更多
关键词 RECEPTOR protein chip estrogen - rogen AGING
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副溶血弧菌QsvR ChIP-qPCR方法的建立 被引量:2
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作者 张苗苗 李雪 +2 位作者 陆仁飞 张义全 周敏 《江苏大学学报(医学版)》 CAS 2023年第6期516-520,527,共6页
目的:建立副溶血弧菌QsvR的染色质免疫共沉淀实时定量PCR(ChIP-qPCR)实验方法。方法:通过热激转化和转化结合将标记2×Flag标签的qsvR片段转入qsvR突变株(ΔqsvR)中,阿拉伯糖诱导表达QsvR-2×Flag蛋白,通过甲醛与基因组DNA进行... 目的:建立副溶血弧菌QsvR的染色质免疫共沉淀实时定量PCR(ChIP-qPCR)实验方法。方法:通过热激转化和转化结合将标记2×Flag标签的qsvR片段转入qsvR突变株(ΔqsvR)中,阿拉伯糖诱导表达QsvR-2×Flag蛋白,通过甲醛与基因组DNA进行交联形成2×Flag-QsvR-DNA复合物,将基因组DNA超声裂解为100~1000 bp大小不等的片段,通过特异性抗原抗体反应将蛋白质-DNA复合物沉淀下来,高盐和加热解交联,回收DNA进行qPCR定量DNA,分析副溶血弧菌体内QsvR与靶基因的结合情况。结果:成功构建出实验菌株ΔqsvR/pBAD33-qsvR-2×Flag,以不加阿拉伯糖诱导为对照,经阿拉伯糖诱导菌株的aphA、opaR、exsB和vtrA的免疫共沉淀量明显较高,表明在副溶血弧菌体内QsvR与aphA、opaR、exsB和vtrA具有结合作用。结论:成功建立了副溶血弧菌QsvR的ChIP-qPCR实验方法,可用于原核生物体内蛋白质-DNA相互作用的研究。 展开更多
关键词 chip-qPCR 蛋白质-DNA复合物 副溶血弧菌 QsvR
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Using the SELDI ProteinChip System to Detect Changes in Protein Expression in Vero Cells after Infection
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作者 Zhi-jun LIU Bin WANG Zhi-yong YAN Xu-xia SONG Dong-meng QIAN Zhi-qiang BAI 《中国病毒学》 CSCD 2007年第1期68-73,共6页
人的疱疹单一的病毒(HSV-1 ) 1 引起美容,眼睛,并且 encephalitic 疾病并且与潜伏的感染和癌症被联系。这里,我们开发了由使用 SELDI 蛋白质薄片在在 vitro 有教养的 Vero 房间检测蛋白质表示的变化在蛋白质水平学习 HSV-1 感染的致... 人的疱疹单一的病毒(HSV-1 ) 1 引起美容,眼睛,并且 encephalitic 疾病并且与潜伏的感染和癌症被联系。这里,我们开发了由使用 SELDI 蛋白质薄片在在 vitro 有教养的 Vero 房间检测蛋白质表示的变化在蛋白质水平学习 HSV-1 感染的致病的一个工具。在有为 12, 24 或 48 h 的 HSV-1 和文化的感染以后,房间被收获并且 lysed。IMAC3 数组被用于 SELDI-TOF-MS 在感染前后检测 proteomic 差别。薄片检测了一系列差别表示蛋白质山峰。有趣地,在 16 912 Da 的山峰和 17 581 Da 与 ISG15 的分子的团精确相应,它可以在感染的过程期间参予抗病毒的活动。因此,我们获得了的结果能用作一个基础学习在病毒和它的主人之间的 HSV-1 和相互作用的致病。另外,他们能为 HSV-1 感染的治疗在新治疗学的目标的发现帮助。关键词 SELDI 蛋白质薄片 - Vero 房间 - HSV-1 - 蛋白质表达式 CLC 数字 R373 基础项目:中国(30540075 ) 的国家自然科学基础; 展开更多
关键词 SELDI protein chip Vero Cells HSV-1 protein Expression
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基于蛋白质芯片技术筛选与甲型流感病毒NS1蛋白互作的宿主因子及通路分析
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作者 张宇 张薇 +13 位作者 高双荣 陈梦苹 王雅欣 徐英莉 曹姗 赵荣华 包蕾 李舒冉 孙静 鲍岩岩 耿子涵 郭姗姗 冀祖恩 崔晓兰 《中国药物警戒》 2024年第1期40-44,50,共6页
目的筛选出与流感病毒Non-structural protein 1(NS1)蛋白结合的人类宿主蛋白并加以分析,确定这些结合蛋白富集的方向及关键蛋白,为抗流感病毒的新药研发提供思路。方法将NS1样本、Biotin样本分别与HuProt^(TM)人类蛋白质组芯片进行杂... 目的筛选出与流感病毒Non-structural protein 1(NS1)蛋白结合的人类宿主蛋白并加以分析,确定这些结合蛋白富集的方向及关键蛋白,为抗流感病毒的新药研发提供思路。方法将NS1样本、Biotin样本分别与HuProt^(TM)人类蛋白质组芯片进行杂交孵育,以两重复均满足Z-Score≥3为筛选条件对与NS1蛋白有结合的宿主蛋白进行筛选得到特异性检出蛋白,实验组(NS1蛋白)与对照组(Biotin)比值I Mean_Ratio≥1.4为条件筛选出显著特异性检出蛋白。用检出的195个蛋白进行GO(Biological Process,Molecular Function,Cellular Component)和KEGG_PATHWAY分析,通过蛋白-蛋白相互作用(PPI)及MCODE分析得到关键蛋白。结果获得显著特异性检出蛋白195个,GO分析结果显示这些蛋白主要参与了mRNA加工、RNA结合、蛋白结合,KEGG分析主要富集到RNA降解、氨基酸的生物合成等通路。得到的4个关键蛋白DDX6、HSPD1、PKLR、MTHFD1中DDX6与RNA的合成、翻译等过程相关,而NS1蛋白可以通过调控流感病毒RNA和宿主RNA促进病毒的感染,推测DDX6可能在该过程发挥作用;其他3个蛋白目前虽然没有明确的研究指明其与流感病毒有关系,但是能在其他RNA病毒的感染过程中发挥作用。结论与NS1结合的人类蛋白主要富集到RNA合成、加工、转录等过程中,MCODE分析得到的关键蛋白有潜力成为抗流感病毒新的靶点,但作用机制需要后续实验进行进一步验证。 展开更多
关键词 甲型流感病毒 NS1蛋白 蛋白质芯片 生信分析 宿主因子 通路
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Transplantation of human placental chorionic plate-derived mesenchymal stem cells for repair of neurological damage in neonatal hypoxic-ischemic encephalopathy
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作者 Lulu Xue Ruolan Du +8 位作者 Ning Bi Qiuxia Xiao Yifei Sun Ruize Niu Yaxin Tan Li Chen Jia Liu Tinghua Wang Liulin Xiong 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第9期2027-2035,共9页
Neonatal hypoxic-ischemic encephalopathy is often associated with permanent cerebral palsy,neurosensory impairments,and cognitive deficits,and there is no effective treatment for complications related to hypoxic-ische... Neonatal hypoxic-ischemic encephalopathy is often associated with permanent cerebral palsy,neurosensory impairments,and cognitive deficits,and there is no effective treatment for complications related to hypoxic-ischemic encephalopathy.The therapeutic potential of human placental chorionic plate-derived mesenchymal stem cells for various diseases has been explored.However,the potential use of human placental chorionic plate-derived mesenchymal stem cells for the treatment of neonatal hypoxic-ischemic encephalopathy has not yet been investigated.In this study,we injected human placental chorionic plate-derived mesenchymal stem cells into the lateral ventricle of a neonatal hypoxic-ischemic encephalopathy rat model and observed significant improvements in both cognitive and motor function.Protein chip analysis showed that interleukin-3 expression was significantly elevated in neonatal hypoxic-ischemic encephalopathy model rats.Following transplantation of human placental chorionic plate-derived mesenchymal stem cells,interleukin-3 expression was downregulated.To further investigate the role of interleukin-3 in neonatal hypoxic-ischemic encephalopathy,we established an in vitro SH-SY5Y cell model of hypoxic-ischemic injury through oxygen-glucose deprivation and silenced interleukin-3 expression using small interfering RNA.We found that the activity and proliferation of SH-SY5Y cells subjected to oxygen-glucose deprivation were further suppressed by interleukin-3 knockdown.Furthermore,interleukin-3 knockout exacerbated neuronal damage and cognitive and motor function impairment in rat models of hypoxic-ischemic encephalopathy.The findings suggest that transplantation of hpcMSCs ameliorated behavioral impairments in a rat model of hypoxic-ischemic encephalopathy,and this effect was mediated by interleukin-3-dependent neurological function. 展开更多
关键词 behavioral evaluations gene knockout human neuroblastoma cells(SH-SY5Y) human placental chorionic derived mesenchymal stem cells INTERLEUKIN-3 neonatal hypoxic-ischemic encephalopathy nerve injury oxygen-glucose deprivation protein chip small interfering RNA
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Differential expression of plasma cytokines in sepsis patients and their clinical implications
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作者 Hui-Xiu Liu Yu-Ying Wang Xue-Feng Yang 《World Journal of Clinical Cases》 SCIE 2024年第25期5681-5696,共16页
BACKGROUND Sepsis,which is characterized by acute systemic inflammation and is associated with high rates of morbidity and mortality,presents a significant challenge in health care.Some scholars have found that the se... BACKGROUND Sepsis,which is characterized by acute systemic inflammation and is associated with high rates of morbidity and mortality,presents a significant challenge in health care.Some scholars have found that the sequential organ failure assessment(SOFA)and quick SOFA scores are not ideal for predicting severe sepsis and mortality.Microbial culture takes a long time(2-3 d)and provides no information for early diagnosis and treatment.Therefore,new diagnostic methods for sepsis need to be explored.AIM To assess cytokine levels in the plasma of sepsis patients and identify potential biomarkers for diagnosing sepsis.METHODS Ten sepsis patients admitted to the emergency department within 24 h of onset were enrolled as the observation group,whereas ten noninfected patients served as the control group.Of the 10 noninfected patients,9 hypertension combined with cerebral infarction,1 patients with vertiginous syndrome.Plasma Cytokines were measured using the Bio-Plex Pro^(TM)Human Chemokine Panel 40-plex.Differentially expressed cytokines in plasma of sepsis and nonsepsis patients were analyzed using Gene Ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses.RESULTS Interleukin(IL)-16,granulocyte-macrophage granulocyte-macrophage colony-stimulating factor(GM-CSF),CX3CL1,CXCL9,CXCL16,CCL25,and CCL23 plasma levels were significantly increased in sepsis patients.GO analysis revealed that these cytokines were mainly associated with cellular structures such as intermediates,nuclear plaques,adhesion plaques,lateral plasma membranes,and cell matrix junctions.These genes were involved in various molecular functions,such as cytokine activity,receptor ligand activity,and signal receptor activator activity,contributing to various biological functions,such as leukocyte chemotaxis,migration,and chemotaxis.KEGG analysis indicated involvement in cytokine cytokine receptor interactions,chemokine signaling pathways,virus–protein interactions with cytokines and cytokine receptors,and the tumor necrosis factor signaling pathway.CONCLUSION Elevated serum levels of IL-16,GM-CSF,CX3CL1,CXCL9,CXCL16,CCL25,and CCL23 in sepsis patients suggest their potential as diagnostic biomarkers for sepsis. 展开更多
关键词 SEPSIS CYTOKINES Biomarkers protein chip PATIENTS
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高尿酸血症状态下低度炎症的病理特点研究
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作者 褚梦真 王雨 +4 位作者 林志健 黄晶 刘亚迪 殷晓阳 张冰 《世界中医药》 CAS 北大核心 2024年第13期1881-1888,共8页
目的:探讨高尿酸血症(HUA)状态下低度炎症的病理特点。方法:根据体质量随机将迪法克鹌鹑分为正常组、模型组,每组10只。以普通饲料:酵母浸膏粉=4∶1制备食饵,并以该食饵喂养模型组鹌鹑,正常组鹌鹑则自由饮食饮水。分别于造模第10、20、3... 目的:探讨高尿酸血症(HUA)状态下低度炎症的病理特点。方法:根据体质量随机将迪法克鹌鹑分为正常组、模型组,每组10只。以普通饲料:酵母浸膏粉=4∶1制备食饵,并以该食饵喂养模型组鹌鹑,正常组鹌鹑则自由饮食饮水。分别于造模第10、20、30天检测血清尿酸,血清炎症介质白细胞介素-1β(IL-1β)、IL-33、IL-2、IL-13、IL-8、IL-17、IL-6、IL-10、IL-12/P40、IL-16、IL-21、C反应蛋白(CRP)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、肿瘤坏死因子-α(TNF-α)、趋化因子CC配体2(CCL2)及γ干扰素(IFN-γ)、神经突起生长导向因子2(Netrin-2)、五聚蛋白3(Pentraxin 3),观察各炎症介质强度变化;造模第30天,取鹌鹑肝、回肠、肾各脏器组织,进行HE染色后观察组织病理形态变化;造模第20天,用基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析差异炎症介质功能及相关信号通路;用Pearson相关性分析方法分析差异炎症介质与血清尿酸水平的相关性。结果:与正常组比较,模型组鹌鹑血清尿酸水平高(P<0.05),以血清IL-17、IL-6、IL-33等为主的白细胞介素类,以IL-8、CCL2为主的趋化因子类,IFN-γ、TNF-α、CRP及GM-CSF水平均升高(P<0.05),而IL-13、IL-10水平降低(P<0.05)。造模第20天,GO/KEGG富集分析结果显示,HUA状态下的低度炎症可能是尿酸代谢靶点群,通过IL-17、Janus激酶信号转导和转录激活因子(JAK-STAT)等信号通路激活、细胞因子-细胞因子间相互作用,从而诱导IL-6、TNF-α等炎症介质产生。2组组织病理变化结果显示,与正常组相比,模型组回肠组织黏膜下层可见炎性细胞浸润,肝、肾组织未见明显差异。差异炎症介质与血清尿酸水平的相关性分析结果显示,鹌鹑血清中IL-6、TNF-α、CRP、IL-33、IL-17、IL-8、IFN-γ、CCL2、GM-CSF、IL-1β、IL-2、IL-6水平均与血清尿酸水平正相关,IL-10、IL-13水平与血清尿酸水平负相关。结论:HUA鹌鹑模型存在低度炎症,该低度炎症可能与尿酸代谢靶点群通过IL-17、JAK-STAT等信号通路的激活以及细胞因子间的相互作用,从而调控IL-6、TNF-α等炎症介质的产生有关。 展开更多
关键词 高尿酸血症 低度炎症 病理特点 鹌鹑 炎症介质 蛋白芯片技术 酶联免疫吸附试验法 基因本体/京都基因与基因组百科全书富集分析
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基于流式量子点微球技术的甲乙型流感病毒抗原检测方法的建立和初步应用分析
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作者 夏成静 李宝花 +3 位作者 郭燕妳 周小合 张润玲 牛英波 《现代检验医学杂志》 CAS 2024年第1期126-130,共5页
目的建立基于流式量子点微球技术的甲型流感病毒(FluA)和乙型流感病毒(FluB)抗原联检方法,为常见呼吸道病毒抗原多重检测打下基础。方法分别使用自制的不同量子点编码微球和小鼠抗FluA/FluB核蛋白(NP)单克隆抗体进行偶联,在流式细胞仪... 目的建立基于流式量子点微球技术的甲型流感病毒(FluA)和乙型流感病毒(FluB)抗原联检方法,为常见呼吸道病毒抗原多重检测打下基础。方法分别使用自制的不同量子点编码微球和小鼠抗FluA/FluB核蛋白(NP)单克隆抗体进行偶联,在流式细胞仪上对已知浓度的甲乙型流感病毒抗原分别和同时进行检测,对检测条件进行优化,建立甲乙型流感病毒抗原联检方法,利用此方法对临床样本进行检测,与实时荧光定量PCR法(quantitative real-time PCR,qPCR)进行比对,验证此方法的临床性能。结果建立了甲乙型流感病毒抗原联检方法,该方法的甲乙型流感病毒抗原的检出限分别为26.1 pg/ml和10.7 pg/ml,测量范围均为15.3~250000 pg/ml。对临床样本检测,与qPCR比对一致性良好,阳性符合率为57.4%,阴性符合率为100%,总符合率71.6%,并优于临床常用胶体金试剂(阳性符合率为56.49%,阴性符合率为99.75%)。结论此流式量子点微球多重检测方法可以用于甲乙型流感病毒抗原的联检,其灵敏度较高、特异度好、检测范围广,可以为呼吸道常见病毒多重检测打下良好基础,在临床上具有应用前景。 展开更多
关键词 甲型流感病毒 乙型流感病毒 量子点编码微球 液相蛋白芯片
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贝伐珠单抗靶向治疗卵巢癌患者对肿瘤标志物的影响
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作者 吴铃 叶健文 +1 位作者 林培红 赖雪丹 《中国卫生标准管理》 2024年第6期142-146,共5页
目的探讨贝伐珠单抗靶向治疗卵巢癌对肿瘤标志物的影响。方法选取2019年1月—2021年12月福建医科大学附属福州市第一医院收治的80例卵巢癌患者,根据不同治疗药物进行分组,分别纳入单独行化疗的化疗组和行贝伐珠单抗靶向治疗的靶向组,每... 目的探讨贝伐珠单抗靶向治疗卵巢癌对肿瘤标志物的影响。方法选取2019年1月—2021年12月福建医科大学附属福州市第一医院收治的80例卵巢癌患者,根据不同治疗药物进行分组,分别纳入单独行化疗的化疗组和行贝伐珠单抗靶向治疗的靶向组,每组40例。测评2组治疗前后肿瘤标志物蛋白芯片中糖链抗原125(glycoantigens in protein chips-125,CA-125)、人附睾蛋白4(Human epididymal protein 4,HE4)水平及血管内皮生长因子(vascular endothelial growth factor,VEGF)水平,治疗后客观缓解率、疾病控制率、免疫指标、癌症患者生命质量测定量表(quality of life scale for cancer patients,FACT-G)和抑郁自评量表(self-rating depression scale,SDS)、焦虑自评量表(self-rating anxiety scale,SAS)评分。结果治疗后,靶向组治疗客观缓解率为65.00%,疾病控制率为87.50%,高于化疗组的42.50%、70.00%(P<0.05);靶向组CA-125、HE4、VEGF指标低于化疗组,CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)高于化疗组,FACT-G评分高于化疗组,而SDS、SAS评分低于化疗组,差异有统计学意义(P<0.05)。结论贝伐珠单抗靶向治疗卵巢癌患者可抑制肿瘤新生血管形成和生长,增强治疗效果,改善患者的心理情绪和生活质量。 展开更多
关键词 贝伐珠单抗 靶向治疗 卵巢癌 肿瘤标志物 CA-125 HE4 VEGF
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E3泛素连接酶CHIP与上皮性癌关系研究
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作者 陈万涛 《口腔颌面外科杂志》 CAS 2014年第6期401-404,共4页
泛素化修饰是机体一种非常重要的蛋白质翻译后修饰方式,其广泛参与细胞周期、DNA修复、信号转导、转录调控等生物学过程。近些年,蛋白质泛素化修饰在恶性肿瘤发生和发展中的作用,受到国内外广泛关注。尤其是E3泛素连接酶,它能特异性识... 泛素化修饰是机体一种非常重要的蛋白质翻译后修饰方式,其广泛参与细胞周期、DNA修复、信号转导、转录调控等生物学过程。近些年,蛋白质泛素化修饰在恶性肿瘤发生和发展中的作用,受到国内外广泛关注。尤其是E3泛素连接酶,它能特异性识别作用底物,泛素化修饰的特异性就取决于该连接酶。研究表明,E3泛素连接酶功能异常与恶性肿瘤等多种疾病的病理过程密切相关,因此分析特定的E3连接酶在恶性肿瘤中的作用和机制,有助于加深对恶性肿瘤发生、发展过程中分子机制的认识,为恶性肿瘤相关分子分类和治疗靶点提供实验基础。本文对E3泛素连接酶CHIP的功能,尤其与上皮性癌关系的研究进展进行综述。 展开更多
关键词 E3泛素连接酶 热休克蛋白70碳末端相互作用蛋白(chip) 上皮性癌
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T细胞免疫检测和蛋白芯片技术筛查人类免疫缺陷病毒感染者结核潜伏感染的效果分析
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作者 金先琴 袁翱 《中外医药研究》 2024年第9期156-158,共3页
目的:分析T细胞免疫检测和蛋白芯片技术筛查人类免疫缺陷病毒(HIV)感染者结核潜伏感染的效果。方法:选取2021年8月—2023年8月贵阳市公共卫生救治中心收治的HIV感染者86例作为研究对象。对HIV感染者进行T细胞免疫检测及蛋白芯片技术筛查... 目的:分析T细胞免疫检测和蛋白芯片技术筛查人类免疫缺陷病毒(HIV)感染者结核潜伏感染的效果。方法:选取2021年8月—2023年8月贵阳市公共卫生救治中心收治的HIV感染者86例作为研究对象。对HIV感染者进行T细胞免疫检测及蛋白芯片技术筛查,以结核分枝杆菌培养结果作为“金标准”,比较两种筛查方法检查结核潜伏感染的效能。结果:“金标准”结果显示,86例HIV感染者中66例存在结核潜伏感染。T细胞免疫检测筛查结核潜伏感染的灵敏度、特异度、准确度高于蛋白芯片技术,差异有统计学意义(P<0.05)。结论:T细胞免疫检测筛查HIV感染者结核潜伏感染的效果优于蛋白芯片技术,灵敏度、特异度、准确度更高,值得临床推广。 展开更多
关键词 T细胞免疫检测 蛋白芯片技术 HIV感染 结核潜伏感染
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副监护子CHIP与神经退行性疾病
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作者 程肖蕊 周文霞 张永祥 《生命科学》 CSCD 2006年第2期145-149,共5页
CHIP属于连接酶类,具有E3泛素连接酶活性,参与能量代谢途径和新陈代谢。包括阿尔茨海默病(Alzheimer'sdisease,AD)、帕金森病(Parkinson'sdisease,PD)、亨廷顿病(Huntington'sdisease,HD)等在内的神经退行性疾病的主要病理... CHIP属于连接酶类,具有E3泛素连接酶活性,参与能量代谢途径和新陈代谢。包括阿尔茨海默病(Alzheimer'sdisease,AD)、帕金森病(Parkinson'sdisease,PD)、亨廷顿病(Huntington'sdisease,HD)等在内的神经退行性疾病的主要病理学特征之一——细胞中异常蛋白的聚集,如tau蛋白和α-突触核蛋白等,副监护子CHIP与分子伴侣,如Hsc70/Hsp70、Hsp90等相互作用对这些异常蛋白的产生具有调节作用。最近研究表明,CHIP改变了Hsc70和Hsp90介导调节的信号通路中蛋白折叠和降解的平衡,参与细胞内蛋白质的质量控制;Hsp70/CHIP伴侣系统在tau蛋白生物学和tau蛋白病理学机制中具有重要作用;CHIP可以作为α-突触核蛋白蛋白酶体降解途径和溶酶体降解途径的分子开关。这些研究进展对于进一步揭示神经退行性疾病的发病机制和研制新一代治疗药物具有重要的作用。 展开更多
关键词 chip 神经退行性疾病 异常蛋白 TAU Α-突触核蛋白
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三维培养结肠癌细胞的微流控芯片荧光成像研究
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作者 蔡书祺 郑璐璐 张大伟 《光学仪器》 2024年第2期14-19,共6页
改良了一种微流控芯片,可用于对结肠癌细胞进行三维培养并实现实时荧光成像。在结肠癌细胞内植入内源性的红色荧光蛋白,使用激光共聚焦显微镜对芯片中三维培养的细胞进行成像。通过细胞内部红色荧光蛋白的表达,可以观测到细胞的生长状态... 改良了一种微流控芯片,可用于对结肠癌细胞进行三维培养并实现实时荧光成像。在结肠癌细胞内植入内源性的红色荧光蛋白,使用激光共聚焦显微镜对芯片中三维培养的细胞进行成像。通过细胞内部红色荧光蛋白的表达,可以观测到细胞的生长状态,实现对细胞的实时监测和高分辨率荧光成像。同时,通过免疫荧光染色来表征反映细胞活性的特征蛋白,其荧光强度和蛋白表达呈正相关。研究结果提示,细胞活性相关蛋白的表达受到微环境的影响,其在芯片三维培养中的活性强于二维培养,表明芯片内环境更加接近真实的人体微环境。该方法为进一步探究肿瘤细胞转移机制及相关药物的筛选研究提供了一种新的技术手段及实验平台。 展开更多
关键词 微流控芯片 三维培养 荧光成像 荧光蛋白
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Reverse ChIP:研究DNA-蛋白质相互作用的新方法 被引量:3
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作者 赵明明 齐锦生 栗彦宁 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2010年第5期407-410,共4页
反向染色质免疫共沉淀技术(reverse chromatin immunoprecipitation assay,Reverse ChIP)是一种在体内状态下分析DNA-蛋白质相互作用的新方法.它用特异的核酸探针捕获靶DNA片段及与其相结合的蛋白质,蛋白质用质谱仪检测,以达到确定靶DN... 反向染色质免疫共沉淀技术(reverse chromatin immunoprecipitation assay,Reverse ChIP)是一种在体内状态下分析DNA-蛋白质相互作用的新方法.它用特异的核酸探针捕获靶DNA片段及与其相结合的蛋白质,蛋白质用质谱仪检测,以达到确定靶DNA位点全部相关蛋白质的目的.其可对靶DNA位点相关蛋白质进行全面、系统地鉴定,特别是寻找已知DNA元件相应的调节蛋白.在发现、鉴定靶DNA位点相关蛋白质和研究DNA-蛋白质相互作用中有重要应用价值. 展开更多
关键词 反向染色质免疫共沉淀技术(Reverse chip) DNA-蛋白质相互作用 靶DNA位点相关蛋白质
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益肾降浊化瘀方对肾间质纤维化大鼠肾组织Smad2、CHIP水平的影响
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作者 远方 赵万超 《中国中医药现代远程教育》 2022年第6期144-146,共3页
目的 观察益肾降浊化瘀方对肾间质纤维化大鼠的治疗作用及肾组织中Smad2及CHIP水平的影响。方法 采用腺嘌呤灌胃的方法建立肾间质纤维化大鼠模型,分空白组、模型组、尿毒清组、益肾降浊化瘀方组。造模8周后,采用免疫组化法和Western Blo... 目的 观察益肾降浊化瘀方对肾间质纤维化大鼠的治疗作用及肾组织中Smad2及CHIP水平的影响。方法 采用腺嘌呤灌胃的方法建立肾间质纤维化大鼠模型,分空白组、模型组、尿毒清组、益肾降浊化瘀方组。造模8周后,采用免疫组化法和Western Blot法分别检测肾组织中Smad2及CHIP蛋白的表达。结果 与模型组比较,益肾降浊化瘀方组Smad2及CHIP蛋白表达明显降低(P<0.05)。结论 益肾降浊化瘀方可能通过减低Smad2及CHIP在肾组织的表达,减轻肾间质纤维化,起到延缓慢性肾衰竭的作用。 展开更多
关键词 益肾降浊化瘀方 肾间质纤维化 chip SMAD蛋白 动物实验
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上皮性卵巢癌血清中肿瘤标志物水平及其临床意义 被引量:10
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作者 单冬勇 成树林 +1 位作者 马邺晨 彭红华 《中南大学学报(医学版)》 CAS CSCD 北大核心 2023年第7期1039-1049,共11页
目的:肿瘤标志物已广泛应用于临床。血清CA125检测是目前临床常用的上皮卵巢癌早期筛查和早期诊断方法之一,但使用单一特异性肿瘤标志物诊断上皮性卵巢癌比较困难。本研究采用肿瘤标志物组合检测法,比较各肿瘤标志物单独检测与联合检测... 目的:肿瘤标志物已广泛应用于临床。血清CA125检测是目前临床常用的上皮卵巢癌早期筛查和早期诊断方法之一,但使用单一特异性肿瘤标志物诊断上皮性卵巢癌比较困难。本研究采用肿瘤标志物组合检测法,比较各肿瘤标志物单独检测与联合检测在上皮性卵巢癌诊断中的价值。方法:收集上皮性卵巢癌患者(n=65)和卵巢良性疾病患者(n=29)的临床资料。采用多肿瘤标志物蛋白质芯片检测癌抗原125(cancer antigen 125,CA125)、糖类抗原242(carbohydrate antigen 242,CA242)、甲胎蛋白(alpha-fetoprotein,AFP)、β-人绒毛膜促性腺激素(beta-human chorionic gonadotropin,β-HCG)、癌胚抗原(carcinoembryonic antigen,CEA)、癌抗原199(cancer antigen 199,CA1990)、神经元-特性烯醇化酶(neuron-specific enolase,NSE)、铁蛋白(Ferritin)、癌抗原153(cancer antigen 153,CA153)以及人生长激素(human growth hormone,HGH)的血清水平,比较其在卵巢良恶性肿瘤中的差异;χ2检验分析肿瘤标志物与卵巢上皮癌患者临床病理特征的相关性;Spearman秩关性分析上皮性卵巢癌CA125与其他肿瘤标志物表达水平及年龄与上述10种肿瘤标志物的相关性;采用灵敏度、特异度、阳性预测值、阴性预测值、约登指数及诊断效率分别评价各肿瘤标志物单项检测及联合检测的诊断价值。结果:上皮性卵巢癌组中的β-HCG、NSE、CA153和CA125水平均高于卵巢良性疾病组。上皮性卵巢癌患者血清中NSE的水平与患者的临床分期相关。此外,上皮性卵巢癌患者血清中CA242、β-HCG、CEA、NSE、Ferritin、CA153水平与CA125水平呈正相关(分别rs=0.497、P<0.001;rs=0.612、P<0.001;rs=0.358、P=0.003;rs=0.680、P<0.001;rs=0.322、P=0.009;rs=0.609、P<0.001),β-HCG、Ferritin、CA153水平与患者年龄呈正相关(分别rs=0.256、P=0.040;rs=0.325、P=0.008;rs=0.249、P=0.046)。在上皮性卵巢癌诊断中,CA125单独检测的灵敏度、约登指数及诊断效率均高于其他9种肿瘤标志物单独检测的结果;CA153、CA199、CA242、Ferritin、CEA与CA125进行联合检测时,其灵敏度均高于CA125单独检测的灵敏度,其中CA125+CEA、CA125+Ferritin+CEA这2种组合的灵敏度分别为89.2%和90.8%,诊断效率均为84.1%,高于其他组合的诊断效率。CA125+CEA联合检测时其约登指数为0.616,高于其他组合。结论:CA125在诊断上皮性卵巢癌方面具有较高的诊断价值。血清中的肿瘤标志物组合检测在上皮性卵巢癌中具有更高的灵敏度和特异度。 展开更多
关键词 上皮性卵巢癌 肿瘤标志物 蛋白质芯片化学发光 联合评估 诊断价值
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