1.Effect of AAVC-I on the migration of human umbilical vein endothelial cells.
Shu LI ; Xin JIN ; Xue-feng LONG ; Jin-li JIA ; Gen-bao ZHANG ; Yun HONG
Chinese Journal of Applied Physiology 2015;31(5):407-410
OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.
METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.
RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.
CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.
Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger
2.The management of thyroid well-differentiated carcinoma invading the upper aerodigestive tract.
Jian XU ; Rong-ning YANG ; Xian-jie ZENG ; Shu-lan LI ; Jian-bo YANG ; Li-gen MO
Chinese Journal of Oncology 2003;25(5):501-503
OBJECTIVETo investigate the management and prognosis of thyroid well-differentiated carcinoma invading the upper aerodigestive tract.
METHODSA retrospective analysis of the management was performed done 62 patients with thyroid well-differentiated carcinoma invading the upper aerodigestive tract. The main method of surgery was shaving excision, and the other means including partial thyrochondrectomy, total laryngectomy, sleeve tracheal resection, sternocleidomastoid myoperiosteal flap and myodermal flap reconstruction, or simply palliative excision. Some patients received postoperative radioactive isotope therapy and radiotherapy. All patients were followed-up for 2 to 15 years with an average of 6.5 years.
RESULTSThe best curative effect was proved in the patients with local invasion, with the lumen uninvolved and their locoregional control rate was 100.0% (17/17). And the second choice was in patients with more extensive involvement of the upper aerodigestive tract structures. For them, extensive surgical management was done attempting to remove all gross disease followed by reconstruction, their locoregional control rate was 87.5% (7/8). And the third place was designated to patients with local invasion for which shaving excision was performed even though minor residual disease was left, their locoregional control rate was 55.6% (5/9). The poorest result went to simple palliative excision. For 17 patients with minor residual tumor, the locoregional control rate of those who were given postoperative radioactive isotope therapy was significantly higher than those without.
CONCLUSIONAccording to the limits and degree of invasion in the upper aerodigestive tract by thyroid well-differentiated carcinoma, different ways of surgery is indicated. For patients with residual disease, radioactive isotope therapy should be used to improve the result and life quality. Advanced lesions should be given postoperative radio therapy.
Adolescent ; Adult ; Aged ; Combined Modality Therapy ; Digestive System ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Respiratory System ; pathology ; Retrospective Studies ; Survival Rate ; Thyroid Neoplasms ; mortality ; pathology ; therapy
3.Changes of MAPK and Akt signaling pathways in hearts and placentas of aborted fetuses with congenital heart disease.
Jie XU ; Zhong-Zhou YANG ; Shu ZHANG ; Shao-Gen WU ; Ya-Li HU
Chinese Journal of Contemporary Pediatrics 2010;12(5):327-332
OBJECTIVETo study the changes of MAPK and Akt signaling pathways in hearts and placentas of aborted fetuses with congenital heart disease (CHD), and investigate their roles in the pathogenesis of CHD.
METHODSTen aborted fetuses with severe CHD (CHD group) and 7 gestational age-matched non-cardiac malformation aborted fetuses (control group) were enrolled. Western blot analysis was undertaken to assess the expression of p38, p38alpha, p-p38, MEF2, ERK, p-ERK, Akt, p-Akt(Ser473) and p-Akt(Thr308) in left ventricles and placentas of the fetuses, while semi-quantitative reverse transcription polymerase chain reaction analysis was used to detect the expression of p38alpha isoforms mRNA in hearts.
RESULTSCompared with the heart samples of the control group, the protein expression levels of p38 and its alpha isoform in 4 cases, p-p38 in 6 cases, MEF2 in 2 cases, p-ERK in 8 cases, Akt in 4 cases, p-Akt(Ser473) and p-Akt(Thr308) in 8 cases decreased, while the protein expression levels of p-p38 in 2 cases and p-Akt(Thr308) in 1 case increased. P-p38 protein level in 3 cases and p-ERK protein level in 2 cases decreased in placentas compared with the control group. The changes of protein expression of MAPK and Akt signaling pathway in hearts were not consistent with those in placentas in the CHD group. The expression of p38alpha isoform2 mRNA showed descent tendency in 4 heart samples with CHD, while the expression of other three p38alpha isoforms mRNA was reduced in only 1 sample compared with the control group.
CONCLUSIONSDysfunction of MAPK and Akt signaling pathways is tissue-specific in aborted fetuses with CHD. The perturbed two signaling pathways in hearts may contribute to the pathogenesis of human CHD.
Female ; Fetus ; metabolism ; Heart Defects, Congenital ; metabolism ; Humans ; MAP Kinase Signaling System ; physiology ; Myocardium ; metabolism ; Phosphatidylinositol 3-Kinases ; physiology ; Placenta ; metabolism ; Pregnancy ; Proto-Oncogene Proteins c-akt ; physiology ; Signal Transduction ; physiology ; p38 Mitogen-Activated Protein Kinases ; physiology
4.Establishment and application of high throughput screening model for hepatitis C virus NS3-4A protease inhibitors in vitro.
Li JIAN-RUI ; Wu YAN-BIN ; Si SHU-YI ; Chen HONG-SHAN ; Jiang JIAN-DONG ; Bang ZONG-GEN
Acta Academiae Medicinae Sinicae 2011;33(1):98-101
OBJECTIVETo establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro.
METHODSHCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated.
RESULTSHigh throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L.
CONCLUSIONThe assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.
Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Fluorescence Resonance Energy Transfer ; Hepacivirus ; enzymology ; High-Throughput Screening Assays ; methods ; Protease Inhibitors ; pharmacology ; Viral Nonstructural Proteins ; antagonists & inhibitors ; genetics
5.Effects of Rapamycin on angiogenesis and tumor progression in human hepatocellular carcinoma implantation mice.
Jian ZHANG ; Hua LI ; Gen-shu WANG ; Nan JIANG ; Yang YANG ; Gui-hua CHEN
Chinese Journal of Surgery 2009;47(18):1403-1405
OBJECTIVETo study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice.
METHODSTumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA.
RESULTS(1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05).
CONCLUSIONRPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.
Animals ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Nude ; Neovascularization, Pathologic ; drug therapy ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; genetics ; Sirolimus ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Xenograft Model Antitumor Assays
6.Effect of lumiracoxib on proliferation and apoptosis of human nonsmall cell lung cancer cells in vitro.
Ji-qing HAO ; Qi LI ; Shu-ping XU ; Yu-xian SHEN ; Gen-yun SUN
Chinese Medical Journal 2008;121(7):602-607
BACKGROUNDLumiracoxib is a highly selective cyclooxygenase-2 (COX-2) inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms.
METHODSThe antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Drug-drug interactions were analyzed using the coefficient of drug interaction (CDI) to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry.
RESULTSLumiracoxib (15 - 240 micromol/L) has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the IC50 values of 2597 micromol/L and 833 micromol/L, respectively. The synergistic effect was prominent when lumiracoxib (15 - 240 micromol/L) was combined with docetaxol (0.2 - 2 micromol/L) (CDI < 1). Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/G1-phase cells and a decrease of S-phase cells.
CONCLUSIONSLumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.
Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Diclofenac ; analogs & derivatives ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Taxoids ; pharmacology
7.Advances in studies on chemical components and pharmacology of epiphytic type medicinal plants in the Orohid family.
Shu LI ; Chun-Lan WANG ; Shun-Xing GUO ; Pei-Gen XIAO
China Journal of Chinese Materia Medica 2005;30(19):1489-1496
Many of the epiphytic Orchids are used as traditional Chinese medicine. The chemical components and pharmacology have been studied in recent 15 years. This article reviewed the studies which will be beneficial to reveal the relatives among these medicinal plants in the Orchid Family and be helpful to develop new drugs.
Adjuvants, Immunologic
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isolation & purification
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pharmacology
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Animals
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Antineoplastic Agents, Phytogenic
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isolation & purification
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pharmacology
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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pharmacology
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Flavones
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isolation & purification
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pharmacology
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Humans
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Molecular Structure
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Orchidaceae
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chemistry
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classification
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Plants, Medicinal
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chemistry
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classification
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Platelet Aggregation Inhibitors
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isolation & purification
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pharmacology
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Polysaccharides
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isolation & purification
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pharmacology
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Stilbenes
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isolation & purification
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pharmacology
8.Mitochondrial mechanisms of apoptosis of human leukemia K562 cells induced by AVVC-1.
Ru-Qi ZHENG ; Gen-Bao ZHANG ; Lu HUANG ; Kai-Ran MA ; Juan WU ; Shu LI
Journal of Experimental Hematology 2013;21(3):591-595
This study was purpose to investigate apoptosis pathway of leukemia K562 cells induced by anticoagulant fraction from Agkistrodon acutus venom (AVVC-1). The mitochondrial transmembrane potential (ΔΨm) of leukemia K562 cells was detected by flow cytometry with JC-1 single staining. The expression of cytochrome C in the mitochondrial of leukemia K562 cells was analyzed by Western blot after AVVC-1 treatment. The distribution of cytochrome C in leukemia K562 cells was measured by immuno-fluorescence test. The results showed that the potential of mitochondrial membrane decreased after treatment with different concentrations of AVVC-1 (12.5, 25, 50, 100 µg/ml) for 6 h (P < 0.01). The expression level of cytochrome C protein in mitochondria obviously declined after treatment with 30 µg/ml AVVC-1 for 48 h, and the fluorescent intensity of cytochrome C in cytosol was enhanced at the same time. It is concluded that AVVC-1-induced K562 cell apoptosis is related with mitochondrial damage, and cytochrome C may be a useful agent for investigating human leukemia therapy by using AVVC-1.
Agkistrodon
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Animals
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Apoptosis
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drug effects
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Cytochromes c
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metabolism
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Humans
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K562 Cells
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Membrane Potential, Mitochondrial
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drug effects
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Mitochondria
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metabolism
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Snake Venoms
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pharmacology
9.Effects of sirolimus on the growth of transplanted hepatocellular carcinoma.
Jian ZHANG ; Hua LI ; Gen-shu WANG ; Nan JIANG ; Yang YANG ; Gui-hua CHEN
Chinese Journal of Hepatology 2009;17(6):413-416
OBJECTIVETo study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice.
METHODSHepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay.
RESULTSThe tumor weight was (352+/-38) mg, (683+/-53) mg and (675+/-45) mg in SRL, FK506 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.01), and there was no difference between FK506 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01).
CONCLUSIONSRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.
Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Immunohistochemistry ; Liver ; blood supply ; pathology ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Proliferating Cell Nuclear Antigen ; metabolism ; Sirolimus ; administration & dosage ; pharmacology ; Tacrolimus ; administration & dosage ; pharmacology ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays
10.Research on HBV DNA inhibition of plasmid acute infection mouse with betulinic acid.
Bing QIAO ; Yue-Qiu GAO ; Man LI ; Shao-Fei WU ; Chao ZHENG ; Shu-Gen JIN ; Hui-Chun WU ; Zhuo YU ; Xue-Hua SUN
China Journal of Chinese Materia Medica 2014;39(6):1097-1100
Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease
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Animals
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Antiviral Agents
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pharmacology
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DNA Replication
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drug effects
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DNA, Viral
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biosynthesis
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Hepatitis B
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blood
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virology
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Hepatitis B Surface Antigens
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blood
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Hepatitis B virus
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drug effects
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genetics
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immunology
;
physiology
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Male
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Mice
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Plasmids
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genetics
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Triterpenes
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pharmacology
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Virus Replication
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drug effects