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Effect of Tongluojiunao injection made from Sanqi(Radix Notoginseng) and Zhizi(Fructus Gardeniae) on brain microvascular endothelial cells and astrocytes in an in vitro ischemic model

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Journal of Traditional Chinese Medicine 2014年第6期34卷 725-732页

作者： Weihong li Xingguang li Qinghong Du Feng li Yuan zhu Yang liu Jie Ma liangqin Wan fanghe li Sai Zhang School of Preclinical Medicine Beijing University of Chinese Medicine Encephalopathy Center Jiangsu Province Hospital of Traditional Chinese Medicine Medical Affairs Novartis Pharmaceuticals (China)

OBJECTIVE: To explore the effect of Tongluojiunao injection(TLJN) prepared with Sanqi(Radix Notoginseng) and Zhizi(Fructus Gardeniae) on the interaction between brain microvascular endothelial cells(BMECs) and astrocytes in an in vitro ischemic ***: First, an in vitro model of cerebral ischemia in BMECs or astrocytes was established by oxygen-glucose deprivation(OGD). TLJN was used as a medicine of intervention. The OGD-injuredBMECs were cultured in various astrocyte-conditioned media. Cell activity, alkaline phosphatase(AKP) and γ-glutamyl transpeptidase(γ-GT) activity,interleukin-1 beta(IL-1β), and tumor necrosis factor alpha(TNF-α) content in BMECs were ***, OGD-injured astrocytes were cultured in various BMEC-conditioned media. Cell activity, as well as expression of brain-derived neurotrophic factor(BDNF) and glial cell-derived neurotrophic factor(GDNF) in astrocytes, were ***: The results of paracrine signaling of normal BMECs or astrocytes showed a protective effect on each other: conditioned media from normal astrocytes improved cell viability, AKP, and γ-GT activity, and reduced IL-1β and TNF-α content of injured BMECs; conditioned media from normal BMECs improved cell viability and expression of BDNF and GDNF in injured astrocytes. However, once the BMECs or astrocytes were injured by OGD, the protective effect decreased or disappeared. The above-mentioned protective induction was effectively recovered by TLJN ***: The therapeutic benefit of TLJN was achieved by recovering two-way induction between BMECs and astrocytes, enhancing activity of injured BMECs and astrocytes, stabilizing enzymatic barriers, promoting expression of neurotrophic factors, and inhibiting inflammatory cytokines.

关键词：脑微血管内皮细胞星形胶质细胞模型制作脑缺血胶质细胞源性神经营养因子注射液体外 lial

Preparation and identification of monoclonal antibodies against the adenovirus vector

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The Chinese-German Journal of Clinical Oncology 2007年第4期6卷 411-414页

作者： Chunyan Zhang Jinsong Gong Yan Chen fanghe li Experimental Medicine Center Tongji Hospital Tongji Medical College Huazhong University of Science andTechnology Wuhan 430030 China

Objective:To prepare and identify monoclonal antibodies(McAbs)against the capsid proteins of adenovirus ***:BALB/c mice were immunized with a mixture of the purified adenovirus vector(Adv)and Al(OH)*** were produced using cell fusion technique in a conventional *** sensitivity and specificity of monoclonal antibodies was identified by indirect enzyme linked immunosorbent assay(EliSA),immunocytochemical staining and Western blotting. Results:Six strains of hybridoma cells(A4H11,A8C7,F1H5,G1D2,G4E3 and H2G8)that can stably secrete the IgG1 McAb against Adv were *** 3 months subculture and low concentration of serum adapting culture,six strains retained their stability to secrete *** ascites titers were between 1:106 and 1:*** blot analysis demonstrated that all the McAbs reacted with one protein(about 114 kDa)which is present in wild type 3 adenovirus(wtAd3),wild type 5 adenovirus (wtAd5),wild type 7 adenovirus(wtAd7)and adenovirus ***:Successfully prepared six strains of hybridoma cell secreted monoclonal antibodies against the hexon proteins of adenovirus vector,and provided the substantial foundation of preclinical research of adenovirus vectors.

关键词：抗腺病毒载体单克隆抗体制备初步鉴定

Preparation, identification, and clinical application ofanti-HBs monoclonal antibody that binds both wild-typeand immune escape mutant HBsAgs

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Frontiers of Medicine 2009年第3期3卷 277-283页

作者： fanghe li Chunyan ZHANG Jinghua liU Xiaoyan ZHANG Bing YAN Bo ZHANG Yongguo HUANG Jingsong GONG Yan CHEN Experimental Center of Medical Sciences Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

Using a standard cellular fusion technique and indirect enzyme-linked immunosorbent assay(EliSA),a hybridoma cell line strain secreting anti-HBs monoclonal antibody(mAb)(deﬁned G6 mAb)was *** cells grew and secreted mAb *** titers in the culture supernatant and ascites were 2.048�106 and 4.096�106,*** applying the anti-HBs G6 mAb and horseradish peroxidase(HRP)-labeled goat anti-HBs antibody,we developed a sandwich EliSA(deﬁned G6m EliSA)for detecting both wild-type and immune escape mutant HBsAgs(IEM HBsAg).The assay was performed to detect 17 species of genome recombinant expression HBsAg,including two wild-type species and 15 IEM HBsAg species,which varied in the“a”determinant,in a group of patients infected with hepatitis B virus(HBV).The patients previously had a lower EliSA detection signal[(absorbance of patients/absorbance of normal people(P/N):1.0–4.5)].The results demonstrated that the sensitivity of this assay to wild-type HBsAg was no less than 0.125μg/L;12 of 15 IEM HBsAg species(P/N≥2.5)were positive for G6 *** the positive IEM HBsAg species,two had a low absorbance value at 450 nm(A450),one had an intermediate A450 value and nine had a high A450 value,which was 7.55%(mean),59.4%and 92.1%–109.4%of the wild-type A450 value,*** two species with low A450 value and the three negative species mutated at the bases 120–124 in theﬁrst loop of the HBV“a”*** the G6 EliSA and two commercial EliSA kits(A and B),177 patients were *** G6 EliSA had a signiﬁcantly higher detection rate than either commercial EliSAs(19.21%vs 14.89%and 6.21%,respectively;P<0.01,P<0.05,respectively).

关键词： hepatitis B virus gene variation hepatitis B surface antigen immune escape enzyme-linked immuno-sorbent assay

Effect of Yiqihuoxue prescription on myocardial energy metabolism after myocardial infarction via cross talk of liver kinase B1-depen-dent Notch1 and adenosine 5'-monophosphate-activated protein kinase

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Journal of Traditional Chinese Medicine 2017年第3期37卷 378-386页

作者： Wu Jiangong Chen Xi Guo Shuwen liu Wenchen Zhang Lu li fanghe Wu Jiani Huang Xiaolou Cai Qian Tan Xiaobo Wang Hui Institute of Traditional Chinese Medicine Beijing University of Chinese MedicineBeijing 100029China

OBJECTIVE: To investigate the effect of Yiqihuoxue prescription(YQHX) from Traditional Chinese Medicine(TCM) on myocardial glucose and lipid metabolism after myocardial infarction via the cross talk between the liver kinase B1(LKB1)-dependent Notch1 and adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK). YQHX was prepared with substances with properties that benefit, to activate blood circulation based on the TCM ***: Animal models of myocardial infarction were established by ligating Sprague Dawley rats' left anterior descending coronary arteries. The animals were randomly divided into a myocardial infarction(MI) group, a YQHX group, a perindopril group, a γ-secretase inhibitor, Notch signal inhibitor(DAPT) group, a DAPT+YQHX group and a sham group. The related drugs were administered on the second day after operation, and changes in the relevant indexes were examined on weeks 1 and *** in cardiac structure and function were examined by echocardiography. The glucose and free fatty acids(FFA) were examined by EliSA. The expression of Notch, LKB1 and AMPK m RNA was examined by a real-time fluorescence quantitative method. The expression of glucose transporter 4(GLUT4), and the expression of total acetyl-Co A carboxylase(ACC) and its phosphorylation were examined by western ***: Compared with the sham group, the expression of Notch, LKB1 and AMPK m RNA in the MI group was lower. Compared with the MI group, the expression of these m RNAs in the YQHX and perindopril groups was higher, and their expression in the DAPT group was lower. At all time points, the protein expression of GLUT4 and p ACC decreased in the MI group. On week 1, the expression of p ACC protein was higher. In the DAPT group, the expression of p ACC protein decreased. Compared with the YQHX group, the expression of p ACC protein in the DAPT + YQHX group was lower. On week 4,compared with the MI group, the expression of GLUT4 protein in the YQHX group and the pe

关键词：心肌梗死蛋白激酶磷酸活化能量代谢活血益气葡萄糖转运蛋白腺苷

Cardioprotective effect of Yiqi Huoxue granule through regulation of mitophagy after myocardial infarction in rats

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Journal of Traditional Chinese Medical Sciences 2020年第2期7卷 171-180页

作者： Xi Chen Jiangong Wu Shuwen Guo Hanrong Mo Tianwei Zhan fanghe li Hui Wang Pengfei Feng Lulu Wei School of Traditional Chinese Medicine Beijing University of Chinese MedicineBeijing100029China Department of Traditional Chinese Medicine Peking University People’s HospitalBeijing100044China Fangshan Hospital Beijing University of Chinese Medicine Beijing102499China

Objective:To investigate the cardioprotective effect of Yiqi Huoxue granule(YQHXG)in the regulation of autophagy in rats induced with myocardial infarction(MI).Methods:An acute MI animal model was established by ligation of the left anterior descending branch of the coronary artery in Sprague-Dawley ***,20 rats received sham operation were classified into a control *** remaining 59 rats were randomly divided into MI model group(n=19),YQHXG group(n=20),and perindopril group(n=20).Relevant indicators on days 7 and 28 were observed in each *** ventricular function was determined by *** structure and morphology of mitochondria,and the number of autophagic vesicles,were observed by transmission electron *** mRNA and protein expression levels of LC3,FUNDC1,Beclin-1,and BNIP3 were examined in the tissue of the MI marginal ***:Compared with the MI model group,YQHXG showed obvious improvements in cardiac *** the microscopic morphology of the heart tissue,myocardial tissue damage attenuated,autophagic signs of autophagosomes and autolysosomes reduced,vacuolization in mitochondria mitigated,and mitochondria arranged in *** could reduce the degree of tissue lesion after MI and regulate the expression of autophagy-related molecules at different *** Day 7,YQHXG significantly downregulated the expression of Fundc1,Becn1,Bnip3 mRNA and reduced the levels of FUNDC1,Beclin-1,BNIP3,and LC3 B proteins expression(all P<.001).On Day 28,YQHXG could upregulate the expression of Becn1,Fundc1 and Bnip3 mRNA and increased the levels of the corresponding proteins expression(all P<.001).Besides,it also increased LC3 B protein expression level(P=.0344).Conclusion:YQHXG regulated the expression of mitochondrial autophagy-related factors in myocardial tissue and mitochondrial autophagic activity at different stages to protect the heart following MI.

关键词： Myocardial infarction Yiqi Huoxue granule Mitochondrial autophagy

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心力衰竭气虚证动物模型的建立与评价

中国实验方剂学杂志 2023年第15期29卷 60-67页

作者：于江唐家杨李帅李想普海崟彭煜李芳赫郭淑贞北京中医药大学北京100029

目的:建立心力衰竭气虚证的病证结合小鼠模型,并对其进行评价。方法:将44只昆明种(KM)小鼠随机分为假手术组、模型组、四君子汤加减组(12.89 g·kg^(-1))。模型组及四君子汤加减组给予主动脉弓缩窄术(TAC)处理,假手术组只穿线不缩窄。... 详细信息

目的:建立心力衰竭气虚证的病证结合小鼠模型,并对其进行评价。方法:将44只昆明种(KM)小鼠随机分为假手术组、模型组、四君子汤加减组(12.89 g·kg^(-1))。模型组及四君子汤加减组给予主动脉弓缩窄术(TAC)处理,假手术组只穿线不缩窄。采用超声心动图、病理学检测对心力衰竭进行疾病模型判断及药效学评估;从宏观表征、微观生物学、方剂反证3个层面分别对小鼠的一般性体征、体质量、旷场、抓力、线粒体超微结构等宏、微观表征进行采集,并测定线粒体分裂、融合蛋白表达,以判断证候类型。结果:TAC术后8周,相较于假手术组,模型组心脏左室射血分数(LVEF)显著降低(P<0.01),四君子汤加减可显著改善小鼠LVEF(P<0.05)。心脏苏木素-伊红(HE)染色结果显示,模型组出现炎性细胞浸润及血管壁增厚,四君子加减可显著改善炎性细胞浸润。在6~8周,相较于假手术组及四君子汤加减组,模型组小鼠出现显著的毛发不齐,毛发枯黄、活跃度降低及精神萎靡的情况。同时与假手术组比较,模型组体质量增长值明显降低(P<0.05);与模型组比较,四君子汤加减组体质量增长值明显上升(P<0.05)。在6~8周,与假手术组比较,模型组旷场距离及旷场速度均明显下降(P<0.05),而四君子汤加减组在第8周明显改善旷场距离及旷场速度(P<0.05)。在6~8周,与假手术组比较,模型组抓力最大值明显下降(P<0.05),而四君子汤加减组TAC术后8周抓力最大值明显增加(P<0.05)。腓肠肌透射电镜显示,模型组肌组织基质不均匀,线粒体肿胀,体积变大,基质溶解,嵴缺失,空泡化;四君子汤加减组可改善线粒体肿胀,嵴断裂,基质空化。蛋白免疫印迹法(Western blot)结果显示,与假手术组比较,模型组小鼠腓肠肌分裂蛋白动力相关蛋白1(DRP1)表达显著增高(P<0.01),线粒体融合素1(MFN1)表达明显降低(P<0.05)。同时与模型组比较,四君子汤加减组小鼠腓肠肌分裂蛋白DRP1表达明显降低(P<0.05),MFN1表达显著升高(P<0.01)。结论:TAC术后小鼠6~8周出现显著的气虚证候表现,同时伴随腓肠肌线粒体形态功能异常,而四君子汤加减可显著改善这一过程。

关键词：心力衰竭气虚证动物模型四君子汤

Yiqi Huoxue Decoction attenuates ischemia/hypoxia-induced oxidative stress injury in H9c2 cardiomyocytes

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Journal of Traditional Chinese Medical Sciences 2018年第3期5卷 271-282页

作者： fanghe li Shuwen Guo Hui Wang Xiaolou Huang Xiaobo Tan Qian Cai Qi Zhang Chunguo Wang Jinghong Hu Wangou lin Beijing University of Chinese Medicine Beijing 100029China Fujian Medical University Fuzhou 350004China

Objective:Yiqi Huoxue Decoction (YQHX) has been widely used for clinical treatment of ischemic heart *** oxidative stress plays a key role in the pathogenesis of ischemic heart disease,the function and molecular mechanism underlying antioxidative protective effects of YQHX on H9c2 cardiomyocytes against ischemia/hypoxia (I/H) have yet to be well ***:H9c2 cells were subjected to 12 h of hypoxia with serum-free conditions and then treated with or without YQHX (100-400 μg/mL).Cell viability was examined using a CCK-8 *** dialdehyde (MDA) and superoxide dismutase (SOD) activity were detected using commercial *** reactive oxygen species (ROS) levels and mitochondrial membrane potential were measured using fluorescence microscopy and confocal laser-scanning microscopy,*** details of mitochondria in H9c2 cells were observed using transmission electron *** antioxidative protective pathway was assessed by measuring mRNA and protein expression of Nrf2 and HO-1,as well as AMPK ***:I/H injury gradually induced oxidative *** with YQHX significantly increased cell viability and reversed I/H-induced oxidative stress,including reducing the production of oxidative stress products (ROS and MDA),increasing SOD levels,improving mitochondrial morphology,and increasing mitochondrial membrane *** was also observed to increase I/H-induced expression of Nrf2 and HO-1,and the activation effects of YQHX were blocked by an AMPK *** addition,HPLC analysis showed that YQHX contained two active antioxidative constituents (calycosin and ferulic acid).Conclusion:The results suggest that anti-oxidative effects exerted by YQHX in H9c2 cardiomyocytes may be linked to upregulation of the AMPK-mediated Nrf2/HO-1 pathway.

关键词： Hypoxia Yiqi Huoxue Decoction Chinese medicine Cardiomyocytes Oxidative stress

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高陡岩质边坡生态修复基材模拟试验研究

人民长江 2024年第3期55卷 218-225页

作者：杨继清陈晓雪杨继华李凤刘芳赫云南农业大学建筑工程学院云南昆明650201 中国有色金属工业昆明勘察设计研究院有限公司云南昆明650051 云南农业大学国际学院云南昆明650201

为修复云南省西北部裸露岩质边坡的生态环境,研发了集多功能于一体的生态修复基材。该生态基材分为结构层和面层,以陶粒、复合肥、保水剂等为原材料,分别设计结构层正交试验和面层正交试验,选取发芽率和生长高度作为定量评价植物生长情... 详细信息

为修复云南省西北部裸露岩质边坡的生态环境,研发了集多功能于一体的生态修复基材。该生态基材分为结构层和面层,以陶粒、复合肥、保水剂等为原材料,分别设计结构层正交试验和面层正交试验,选取发芽率和生长高度作为定量评价植物生长情况的指标。根据设计的各试验组配比和观测结果,建立模糊综合评价模型和多项式回归模型,求出其在给定条件下的最优配比,并分析各影响因素间的关系。结果表明:基材结构层最优配比为谷壳750 g/m^(2)、复合肥20 g/m^(2)、EPS颗粒40 g/m^(2)、陶粒1.88 kg/m^(2)、团粒剂34.65 g/m^(2)、保水剂84.6 g/m^(2);基材面层最优配比为复合肥9.24 g/m^(2)、保水剂9.12 g/m^(2)、团粒剂13.51 g/m^(2)。基材各组分存在交互作用,而土壤的理化性质与护坡植物生长存在一定的相关性。研究结果可为高寒地区高陡岩质边坡生态基材的制备与生态修复提供理论依据和技术支持。

关键词：岩质边坡生态修复基材正交试验多项式回归

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乙肝病毒抗原多肽的印迹法分析

中华实验和临床病毒学杂志 1992年第4期6卷 385-389页

作者：郭林生杨东亮同济医科大学附属同济医院，武汉 430030

采用印迹法（Western Blot）技术对血源性Dane颗粒的抗原性多肽进行了多克隆和单克隆抗体分析。结果表明：①P 24P 27，P 36和P 39P 42都是S-ORF区的多肽，分别含有S区，S 2区和S 1区特异性结构位... 详细信息

采用印迹法（Western Blot）技术对血源性Dane颗粒的抗原性多肽进行了多克隆和单克隆抗体分析。结果表明：①P 24P 27，P 36和P 39P 42都是S-ORF区的多肽，分别含有S区，S 2区和S 1区特异性结构位点，多克隆抗体识别提示三者之间存在有 "重复抗原决定簇" ；②多克隆抗HBe和抗HBc均能同P 21结合，推测该多肽可能是来自C区的具有e抗原和核心抗原两种抗原位点的多肽；③兔抗Pol能在P 24- 25处特异性呈色，说明这一多肽不仅含有S区的基因产物、而且可能还含有与抗Pol结合的成分。此外，还发现P 45，P 76分别具有与人免疫球蛋白IgG、IgM相似的抗原特性；④采用针对x、o1、e、c产物的多株单克隆抗体识别为阴性，表现出这一技术在单抗识别上有一定的局限性。

关键词：印迹法多肽乙型肝炎病毒抗原