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基于HGF/PI3K/Akt信号轴研究赤芍-附子药对促进慢加急性肝衰竭肝细胞再生的作用机制

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中国中药杂志 2024年第15期49卷 4188-4196页

作者：田晓玲张彧杜珊伍梦思谭年花陈斌湖南中医药大学湖南长沙410208 湖南中医药大学第一附属医院湖南长沙410007

基于肝细胞生长因子(hepatocyte growth factor,HGF)/磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)信号轴研究赤芍-附子药对对慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)大鼠的治疗作用及对肝细胞再生的影响。采用牛血清白蛋白皮下及尾静脉注射联合脂多糖(lipopolysaccharides,LPS)+D-氨基半乳糖(D-galactosamine,D-GalN)腹腔注射构建ACLF大鼠模型,实验设置正常对照(NC)组、模型(vehicle)组、赤芍-附子药对(SFYD,5.85 g·kg^(-1))组、促肝细胞生长颗粒(HGFG,4.05 g·kg^(-1))组。苏木素-伊红(hematoxylin-eosin staining,HE)染色观察大鼠肝组织病理变化。全自动生化分析仪检测血清谷丙转氨酶(alanineamino transferase,ALT)、谷草转氨酶(aspartate transa-minase,AST)、总胆红素(total bilirubin,TBIL)。ELISA法检测白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)炎症因子水平。免疫荧光染色检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、细胞增殖相关抗原(antigen identified by monoclonal antibody,Ki67)及细胞周期蛋白(cell cycle protein B1,CyclinB1)阳性表达。实时荧光定量聚合酶链式反应(rt-qPCR)法、蛋白免疫印迹(Western blot)法检测HGF、结合蛋白Gab1(grb2 associated binding protein 1,Gab1)、PI3K、Akt、磷酸化PI3K(phosphorylation PI3K,p-PI3K)、磷酸化Akt(phosphorylation Akt,p-Akt)mRNA及蛋白表达水平。结果显示,与vehicle组比较,SFYD组大鼠肝组织病理评分降低,肝功能及炎症反应改善,PCNA、Ki67、CyclinB1荧光表达增强。同时与模型组相比,SFYD组HGF、Gab1蛋白表达上调,PI3K、Akt磷酸化激活。以上研究表明赤芍-附子药对通过减轻肝细胞炎症损伤以及促进肝细胞再生来达到抗肝衰竭的效应,其具体调控机制可能与激活HGF/PI3K/Akt信号通路有关。

关键词：慢加急性肝衰竭赤芍-附子药对 HGF/PI3K/Akt 肝再生

Tyrosine Hydroxylase as a Target for Deltamethrin in the Nigrostriatal Dopaminergic Pathway

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Biomedical and Environmental Sciences 2006年第1期19卷 27-34页

作者： GONG-PING LIU QIANG MA nian SHI Department of Toxicology MOE key Laboratory of Environmental and Health Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei China Receptor Biology Laboratory Toxicology and Molecular Biology Branch Health Effects Laboratory Division National Institute for Occupational Safety and Health Centers for Disease Control and Prevention Morgantown WV 26505 USA

Objective To study the effects of deltamethrin on tyrosine hydroxylase in nigrostriatum of male rats. Methods Sprague-Dawley rats were daily treated with deltamethrin at 6.25 or 12.5 mg/kg body weight by gavage for 10 days. Then HPLC-fluorescence detection was used to analyze the contents of dopamine （DA）, 3,4-dihydmxyphenylacetic acid （DOPAC） and homoranillic acid （HVA） in substantial nigra and striatum. The activities of tyrosine hydroxylase （TH） were also detected by HPLC-fluorescence detection. TH mRNA or TH protein levels were measured by rt-PCR and immunohistochemistry method. Results The content of DA in stfiatum was significantly decreased by the treatments, suggesting an inhibition of DA synthesis by deltamethrin. The contents of DA metabolites DOPAC and HVA increased, indicating increased dopamine turnover. Furthermore, deltamethrin significantly decreased the activity, as well as the mRNA and protein levels of TH. Conclusions These findings reveal a novel aspect of deltamethfin neurotoxicity and suggest tyrosine hydroxylase as a molecular target of deltamethin on dopamine metabolism in the nigrostriatal pathway.

关键词： Deltamethrin Nigrostriatum Dopamine Tyrosine hydroxylase Parkinson's disease

LncRNA C9orf139 can regulate the growth of pancreatic cancer by mediating the miR-663a/Sox12 axis

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World Journal of Gastrointestinal Oncology 2020年第11期12卷 1272-1287页

作者： Jin-nian Ge Di Yan Chun-Lin Ge Min-Jie Wei Department of General Surgery The First Affiliated Hospital of China Medical UniversityShenyang 110001Liaoning ProvinceChina Intensive Care Unit The Central Affiliated Hospital of Shenyang Medical CollegeShenyang 110024Liaoning ProvinceChina Department of Pharmacology School of PharmacyChina Medical UniversityShenyang 110122Liaoning ProvinceChina

BACKGROUND Recent studies have proved the important role of many oncogenic long noncoding RNAs(lncRNAs) in the progression of pancreatic cancer, but little is known about the mechanisms of tumor suppression in pancreatic *** To evaluate the function of tumor suppressor lncRNA C9orf139 in pancreatic cancer progression and to study the underlying *** We assigned 54 patients with pancreatic ductal adenocarcinoma treated at our hospital to the patient group and 30 normal subjects undergoing physical examination to the control group. rt-qPCR was used to measure the relative expression of C9orf139 in the tissue and serum of patients, in an attempt to investigate the prognostic value of C9orf139 in pancreatic cancer patients. The luciferase reporter gene assay was performed to determine the interaction between C9orf139 and miR-663 a. The biological function of C9orf139 was assessed by in vitro assays and in vivo subcutaneous tumor formation tests in animal models. To figure out the molecular mechanism of C9orf139 to act on miR-663 a/Sox12, RNA pull-down, Western blot assay, RNA immunoprecipitation assay, and co-immunoprecipitation assay were *** C9orf139 level significantly increased in the tissue and serum of patients, which had clinical diagnostic value for pancreatic cancer. Patients with high C9orf139 expression had a higher risk of progressing to stage Ⅲ + Ⅳ, lymph node metastasis, and poor differentiation. Cox regression analysis suggested that C9orf139, tumor-node-metastasis stage, and lymph node metastasis were independent prognostic factors in patients. The underlying mechanism of C9orf139 was that it promoted the growth of pancreatic cancer cells by modulating the miR-663 a/Sox12 *** C9orf139 is highly expressed in pancreatic cancer, qualified to be used as a potential diagnostic and prognostic marker for pancreatic cancer. Its promotion of pancreatic cancer cell growth is achieved by mediating the miR-663 a/Sox12 ax

关键词： C9orf139 miR-663a Sox12 Pancreatic cancer Prognosis Tumor formation in nude mice

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云南松PyCADs基因的克隆与表达分析

西南农业学报 2024年第6期37卷 1203-1211页

作者：母德锦陈林许玉兰蔡年辉冯玲陈诗唐军荣西南林业大学/西南地区生物多样性保育国家林业和草原局重点实验室昆明650224

【目的】研究云南松肉桂醇脱氢酶的表达特性及其在木质素生物合成过程中的作用。【方法】以一年生云南松苗木为材料,通过测定嫩茎、嫩芽和针叶中18种木质素通路小分子的相对含量,并结合转录组数据联合分析,对云南松CAD基因进行筛选、克... 详细信息

【目的】研究云南松肉桂醇脱氢酶的表达特性及其在木质素生物合成过程中的作用。【方法】以一年生云南松苗木为材料,通过测定嫩茎、嫩芽和针叶中18种木质素通路小分子的相对含量,并结合转录组数据联合分析,对云南松CAD基因进行筛选、克隆和生物信息学分析,同时利用rt-qPCR技术分析其在不同组织中的表达模式。【结果】在检测的18种木质素通路小分子中,嫩芽以5-O-咖啡酰莽草酸、5-O-对香豆酰莽草酸和咖啡酸3种小分子的相对含量较高,嫩茎以3-O-对香豆酰奎宁酸、4-香豆酸、阿魏酸、松柏醛、L-苯丙氨酸、松柏醇、对-香豆醇、L-酪氨酸和芥子醛9种小分子的相对含量较高,而在针叶中则以松柏苷、香豆醛、咖啡醛和紫丁香苷4种小分子的相对含量较高。云南松转录组中筛选出4个CAD家族成员,通过系统进化分析发现云南松PyCAD-3与AtCAD1同源,PyCAD-1、PyCAD-2和PyCAD-4与松属类CAD、拟南芥AtCAD4、AtCAD5亲缘关系最近。相关性分析表明PyCAD-1和PyCAD-2与上下游木质素通路小分子相关性较强。PyCAD-1和PyCAD-2开放阅读框全长分别为570和1083 bp,编码蛋白分子量分别为20.38和39.30 kDa。实时荧光定量PCR结果分析显示,PyCAD-1基因的相对表达量在嫩芽、嫩茎中最高;PyCAD-2基因的相对表达量在嫩茎中最高。【结论】PyCAD-2可能直接参与云南松木质素的生物合成,而PyCAD-1则可能存在一定的功能冗余。研究结果为云南松CAD基因家族及木质素代谢途径相关基因研究提供参考。

关键词：云南松肉桂醇脱氢酶基因克隆表达分析

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Recrystallization temperature of tungsten with different deformation degrees

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Rare Metals 2016年第7期35卷 566-570页

作者： Xiao-Xin Zhang Qing-Zhi Yan Chun-Tian Yang Tong-nian Wang Min Xia Chang-Chun Ge Institute of Nuclear Materials University of Science & Technology Beijing

Pure tungsten （PW） and W-1 wt% La203 （WL10） were prepared by powder metallurgical route fol- lowed by the swaging ＋ rolling process. The logarithmic strains are 0, 0.37, 0.58, and 0.98 for WL10 and 0, 0.58 for PW. Heat treatments were performed at temperatures var- ied from 1,573 to 2,173 K to determine the recrystalliza- tion temperature. Recrystallization temperatures are 1,973 and 2,173 K for WL1 （logarithmic strain of 0.37） and WL3 （logarithmic strain of 0.98）, respectively. But in the case of WL2 （logarithmic strain of 0.58）, full recrystallization is not achieved at temperature of above 2,173 K. Further- more, the recrystallization temperature of PW with loga- rithmic strain of 0.58 is at least 300 K lower than that of the equivalent WLI0 sample. Moreover, the increase of recrystallization temperature inhibits the strength degra- dation of WL2： samples lose 4 % and 22 % strength when annealed at 1,573 and 1,973 K compared with room tem- perature （rt） sample. Finally, the texture evolution for the swaged ＋ rolled WL10 is significantly related to the deformation degree： the dominated orientation is （001） for WL2 while （110） for WL3.

关键词： Tungsten La203 Swaging+rolling Deformation Recrystallization

miR-210通过Wnt/β-catenin促进结直肠癌对顺铂的药物敏感性研究

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遵义医科大学学报 2024年第1期47卷 62-69页

作者：何念曲颜张念杰万磊杨雪峰遵义医科大学第二附属医院胃肠外科贵州遵义563006

目的探讨miR-210对结直肠癌细胞增殖及顺铂(DDP)药物敏感性及其机制影响。方法 rt-qPCR检测人正常结肠上皮细胞(HCoEpiC)、结直肠癌细胞(HCT-116、HT-29、SW620)及顺铂耐药细胞株(HCT-116/DDP)中miR-210的表达;使用CCK-8和Annexin V-FI... 详细信息

目的探讨miR-210对结直肠癌细胞增殖及顺铂(DDP)药物敏感性及其机制影响。方法 rt-qPCR检测人正常结肠上皮细胞(HCoEpiC)、结直肠癌细胞(HCT-116、HT-29、SW620)及顺铂耐药细胞株(HCT-116/DDP)中miR-210的表达;使用CCK-8和Annexin V-FITC/PI法检测miR-210过表达或干扰后,HCT-116细胞增殖及凋亡情况。将HCT-116/DDP细胞随机分成Control组、miR-210 mimics组、miR-210 inhibitor组,在DDP(80μmol/L)或对照作用24 h后,CCK-8检测HCT-116/DDP细胞活力;Annexin V-FITC/PI双染检测细胞凋亡,Western blot检测细胞凋亡相关指标(Bad、Bcl-2、Cleaved Caspase-3)、耐药相关指标(ABCG2、MRP2、P-gp)及Wnt/β-Catenin通路中重要蛋白的表达。结果与HCoEpiC细胞相比,结直肠癌细胞(HCT-116、HT-29、SW620)中miR-210的表达显著降低,而HCT-116/DDP细胞中miR-210的表达比HCT-116细胞中进一步降低。miR-210 mimics能够显著抑制结直肠癌细胞及其顺铂耐药株的细胞活力,促进细胞凋亡,而inhibitor作用相反。在HCT-116/DDP细胞中,与DDP组比较,DDP+mimics联合组能够显著抑制细胞活力,促进细胞凋亡,促进Bad、Cleaved Caspase-3的表达,抑制Bcl-2的表达,抑制耐药相关蛋白(p-gp、MRP2、ABCG2)的表达,抑制Wnt3a、β-Catenin的表达;miR-210 inhibitor则得到相反的结果。结论 miR-210可能通过调控Wnt/β-catenin通路,影响耐药相关蛋白(p-gp、ABCG2、MRP2)的表达,最终促进HCT-116/DDP耐药细胞对顺铂的药物敏感性。

关键词：结直肠癌耐药性细胞凋亡 miR-210 细胞增殖

Fine mapping and genetic analysis of resistance genes,Rsc18,against soybean mosaic virus

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Journal of Integrative Agriculture 2022年第3期21卷 644-653页

作者： LIU Sang-lin CHENG Yan-bo MA Qi-bin LI Mu JIANG Ze XIA Qiu-ju nian Hai The State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources South China Agricultural UniversityGuangzhou 510642P.R.China The Key Laboratory of Plant Molecular Breeding of Guangdong Province College of AgricultureSouth China Agricultural UniversityGuangzhou 510642P.R.China Maize Research Institute Jilin Academy of Agricultural SciencesGongzhuling 130033P.R.China Beijing Genomics Institute(BGI)-Shenzhen Shenzhen 518086P.R.China

Soybean mosaic virus(SMV) affects seed quality and production of soybean(Glycine max(L.) Merr.) ***18 is one of the dominant SMV strains in South China,and accession Zhonghuang 24 displayed resistance to *** F_(1),F_(2) and 168 F_(11) recombinant inbred lines(RILs) population derived from a hybridization between Zhonghuang 24(resistant,R) and Huaxia 3(susceptible,S) were used in this *** to the segregation ratios of the F_(2) generation(3 R:1 S) and the recombinant inbred lines(RILs) population(1 R:1 S),one dominant locus may regulate the resistance to SC18 in Zhonghuang *** using composite interval mapping(CIM),Rsc18 was mapped to a 415.357-kb region on chromosome *** candidate genes,including one NBS-LRR type gene and two serine/threonine protein type genes,were identified according to the genetic annotations,which may be related to the resistance to *** q rt-PCR demonstrated that these genes were up-regulated in the R genotype compared to the *** conclusion,the findings of this research enhanced the understanding about the R genes at the Rsc18 ***,our results will provide insights for designing molecular markers to improve marker-assisted selection and developing new varieties with resistance to SC18.

关键词： soybean mosaic virus(SMV) fine mapping recombinant inbred lines(RILs) resistance gene

意大利蜜蜂AmMETTL3基因的分子克隆与表达模式

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中国农业大学学报 2024年第1期29卷 99-108页

作者：刘小玉刘治滩荆欣冯佩林高旭泽吴鹰李琪明樊念陈大福付中民郭睿福建农林大学动物科学学院(蜂学学院) 福州350002 福建省蜂疗研究所福州350002

为克隆意大利蜜蜂(Apis mellifera ligustica)的AmMETTL3基因,解析AmMETTL3蛋白的分子特性,并测定AmMETTL3在工蜂不同组织及发育时期的表达谱。通过PCR扩增AmMETTL3的CDS并进行Sanger测序验证;使用相关生物信息学软件预测AmMETTL3的理... 详细信息

为克隆意大利蜜蜂(Apis mellifera ligustica)的AmMETTL3基因,解析AmMETTL3蛋白的分子特性,并测定AmMETTL3在工蜂不同组织及发育时期的表达谱。通过PCR扩增AmMETTL3的CDS并进行Sanger测序验证;使用相关生物信息学软件预测AmMETTL3的理化性质和分子特性,鉴定METTL3的结构域和保守基序,并构建系统进化树;采用rt-qPCR检测AmMETTL3在工蜂7个组织及5个不同发育时期的相对表达量。结果表明:成功克隆到AmMETTL3的CDS并发现AmMETTL3不含典型的跨膜结构域和信号肽;METTL3在各种昆虫中高度保守且含1个MT-A70结构域和3个相同的保守基序;AmMETTL3与东方蜜蜂METTL3的亲缘关系最近;相较于触角中的表达量,AmMETTL3的表达量在咽下腺中极显著上调(P<0.01),在中肠中极显著下调(P<0.01);AmMETTL3在3日龄幼虫中的表达量极显著(P<0.01)低于卵中的表达量,在12日龄蛹中的表达量极显著(P<0.01)高于卵中的表达量,在2、6、12、15和18日龄工蜂体内的表达量极显著(P<0.01)低于1日龄工蜂体内的表达量。研究结果为深入探究AmMETTL3的功能及其参与的表观调控机制提供了参考和依据。

关键词：意大利蜜蜂 N6-甲基腺苷甲基化转移酶3 MT-A70 分子特性表达模式

Novel serum microRNAs panel on the diagnostic and prognostic implications of hepatocellular carcinoma

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World Journal of Gastroenterology 2018年第24期24卷 2596-2604页

作者： Yang An Song Gao Wen-Chao Zhao Bao-An Qiu nian-Xin Xia Peng-Jun Zhang Zhen-Ping Fan Department of Hepato-Biliary-Pancreatic Surgey Navy General Hospital of Chinese People’s Liberation Army Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing) Interventional Therapy DepartmentPeking University Cancer Hospital & Institute The Liver Disease Center for Cadre Medical Care Beijing 302 Military Hospital

AIM To determine a panel of serum micro RNAs(mi RNAs) that could be used as novel biomarkers for diagnosis of hepatocellular carcinoma(HCC).METHODS We initially screened 9 out of 754 serum mi RNAs by Taq Man Low Density Array in two pooled samples respectively from 35 HCC and 35 normal controls, and then validated individually by rt-qP CR in another 114 patients and 114 controls arranged in two phases. The changes of the selected mi RNAs after operation and their prognostic value were *** miR-375, miR-10 a, miR-122 and miR-423 were found to be significantly higher in HCC than in controls(P < 0.0001), and the area under the receiver-operating-characteristic curve for the 4-miR NA panel was 0.995(95%CI: 0.985-1). All the four mi RNAs were significantly reduced after surgical removal of the tumors(P < 0.0001), while still higher than normal controls(at least P < 0.05)CONCLUSION The four serum miR NAs(miR-375, miR-10 a, miR-122 and miR-423) could potentially serve as novel biomarkers for the diagnostic and prognostic of HCC.

关键词： hepatocellular carcinoma microRNAs serum biomarker diagnosis