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| 1 | Cloning of HBsAg-encoded genes in different vectors and their expression in eukaryotic cells显示文摘AIM: To compare the efficiency of different plasmids as DNA vectors by cloning three HBsAg-encoded genes into two eukaryotic expression vectors, pRc/CMV and pSG5UTPL/Flag, and to express HBsAg S, MS, and LS proteins in SP2/0 cells, and to establish monoclone SP2/0 cell strains that are capable of expressing S or S2S proteins stably.METHODS: Segments of S, preS2-S, preS1-preS2-S genes of Hepatitis B virus were amplified by routine PCR and preS1S fragment was amplified by Over-Lap Extension PCR. The amplified segments were cleaved with restricted endonuclease Hind Ⅲ/Not Ⅰ followed by ligation with pRc/CMV, or BamHI/EcoR Ⅰ followed by ligation with pSG5UTPL/Flag. After the plasmid vectors were cleaved with the correspond enzymes, the amplified segments were inserted into pRc/CMV or pSGSUTPL/Flag plasmid vectors with T4DNA ligase. KOZAK sequence was added before the initial ATG code of each fragment using specific primer. The inserted segments in the recombinant plasmids were sequenced after subcloning. BALB/c mice myeloma cells (SP2/0 cell line) were transfected with the recombinant plasmids. The expressions of the different recombinants were compared by Western-blot, using a monoclonal anti-HBs antibody as the primary antibody and peroxidase-labeled multi-linker as the secondary. Stable SP2/0-pRc/CMV-S or SP2/0- pRc/CMV-MS clones were established through clone screening with G418.RESULTS: Fragments with anticipated size were harvested after PCR. After recombination and screening, the sequences of the inserted segments in the recombinants were confirmed to be S, preS2S, preSl-preS2S and preSlS encoding genes,determined by sequencing. The results of Western-blot hybridization were positive for the anticipated proteins.Among them, pRc/CMV-S or pRc/CMV-MS demonstrated the highest expressing their respective antigen.CONCLUSION: Eight recombinant plasmids expressing S,M, L or preSlS proteins are obtained. For hepatitis surface antigen expression in eukaryotic cells, the vector pRc/CMV is superior to pSG5UTPL/Flag, and pRc/CMV-S and pRc/CMVMS are the most efficient in the pRc/CMV clones. SP2/0 cells stably expressing HBsAg are established, and may be used as target cells for evaluating the CTL activity of a DNA vaccine in vitro. | ShanQin HongTang Lian-SanZhao FangHe YongLin LiLiu Xiao-MeiHe | 2003 | World Journal of Gastroenterology2003，9，5： | 12 |
| 2 | 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显示文摘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 theory.METHODS: 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 4.Changes 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 blotting.RESULTS: 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 perindo-pril group was higher. The expression of GLUT4 protein in the DAPT group decreased. Compared with the YQHX group, the expression of GLUT4 protein in the DAPT+YQHX group was lower. There was no significant difference in the expression of ACC protein between the groups.CONCLUSION: YQHX promoted cross talk between the LKB1-dependent Notch1 and AMPK in myocardial tissue after myocardial infarction. Furthermore,it regulated the glucose and lipid metabolism of cardiomyocytes at different time points, thereby ameliorating the cardiac energy metabolism via different mechanisms and protecting the heart. | Wu Jiangong Chen Xi Guo Shuwen Liu Wenchen Zhang Lu Li Fanghe Wu Jiani Huang Xiaolou Cai Qian Tan Xiaobo Wang Hui | 2017 | Journal of Traditional Chinese Medicine2017，37，3： | 7 |
| 3 | 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显示文摘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 model.METHODS: 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 determined.Additionally, 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 detected.RESULTS: 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 intervention.CONCLUSION: 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. | Weihong Li Xingguang Li Qinghong Du Feng Li Yuan zhu Yang Liu Jie Ma Liangqin Wan Fanghe Li Sai Zhang | 2014 | Journal of Traditional Chinese Medicine2014，34，6： | 2 |
| 4 | Landfill leachatetreatmentwithcombinedUASB and Fenton coagulation显示文摘 | WANG P LAU I W C FANGH H P | 2000 | Journal of EnvironmentalScience andHealth2000，35，10： | 1 |
| 5 | Matrine inhibits invasiveness and metastasis of human malignant melanoma cell llne A375in vitro显示文摘 | Liu XY FangH Yang ZG | 2008 | International J dernlato2008，47，5： | 1 |
| 6 | A new ultrasonic de vice in maintenance therapy : perception of pain and clinical effi- eacy显示文摘 | Kocher T Fangh/inel J Schwahn C | 2005 | J Clin Periodontol2005，32，4： | 1 |
| 7 | Cardioprotective effect of Yiqi Huoxue granule through regulation of mitophagy after myocardial infarction in rats显示文摘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 rats.Besides,20 rats received sham operation were classified into a control group.The 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 group.Left ventricular function was determined by echocardiography.The structure and morphology of mitochondria,and the number of autophagic vesicles,were observed by transmission electron microscopy.The mRNA and protein expression levels of LC3,FUNDC1,Beclin-1,and BNIP3 were examined in the tissue of the MI marginal area.Results:Compared with the MI model group,YQHXG showed obvious improvements in cardiac functions.Observing 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 order.YQHXG could reduce the degree of tissue lesion after MI and regulate the expression of autophagy-related molecules at different stages.On 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. | Xi Chen Jiangong Wu Shuwen Guo Hanrong Mo Tianwei Zhan Fanghe Li Hui Wang Pengfei Feng Lulu Wei | 2020 | Journal of Traditional Chinese Medical Sciences2020，7，2： | 1 |
| 8 | Effects of a single inhalative exposure to formaldehyde on the open field behavior of mice显示文摘 | Fathi A. Malek Klaus-Uwe M?ritz Jochen Fangh?nel | 2004 | International Journal of Hygiene and Environmental Health2004，，2： | 1 |
| 9 | Problems of supernumerary teeth,hyperdontia or dentes supernumerarii 显示文摘 | Proff P Fanghä nel J Allegrini S Jr | 2005 | Ann Anat2005，188，2： | 1 |
| 10 | Vitamin D in SLE:modest association with disease activity and urine protein/creatinine ratio 显示文摘 | Etri M BelloKj FangH | 2013 | Arthritis Rheum2013，65，7： | 1 |
| 11 | Clinical interaction of common and important drugs 显示文摘 | FANGH ZENGHZ | 2013 | 中国医院用药评价与分析2013，13，3： | 1 |
| 12 | Effects of a single inhalative exposure to formaldehyde on the open field behavior of mice显示文摘 | Malek FA M(o)ritz KU Fangh(a)nel J | | 0，，02： | 1 |
| 13 | Dietary fiber intake and risk of type 2 diabetes: a dose-response analysis of prospective studies显示文摘 | YaoB FangH XuW eta2 | 2014 | Eur J Epidemiol2014，29，： | 1 |
| 14 | Seizures after primary in-tracerebral hemorrhage显示文摘 | Fanghe E Peter D Bartoluai A | 1989 | Neurology1989，39，： | 1 |
| 15 | Inducedpluripotentstemcellsforspinalcordinjurytherapy:currentstatusandperspective显示文摘 | WangH FangH DaiJ etal | 2013 | NeurolSci2013，34，1： | 1 |
| 16 | StudyonmechGanismoffrictionmatrixcompositesandwearofaluminumitsapplications显示文摘 | OuyangQB FangH WangWL etal | 2004 | AutomobileTechnologyAnd MateriGal2004，，7： | 1 |
| 17 | The morphological and clinical relevance of mandibular and maxillary bone structures for implantation显示文摘 | Fangh(a)nel J Proff P Dietze S | 2006 | Folia Morphol (Warsz)2006，65，1： | 1 |
| 18 | Expressionsofhippocampalmineralocorticoidreceptor(MR)andglucocorticoidreceptor(GR)inthesingle-prolongedstess-rats显示文摘 | ZheD FangH YuxiuS | 2008 | ActaHistochemCytochem2008，41，4： | 1 |
| 19 | Charac terization of microbial community in granular sludge treating brewery wastewater 显示文摘 | LIUWT CHANOC FANGH H P | 2002 | Water Research2002，36，7： | 1 |
| 20 | Silver catalysis in the fabrication of silicon nanowire arrays显示文摘 | FangH Wu Y Zhao J H | 2006 | Nanotechnology2006，17，15： | 1 |
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