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1.
Annals of Dermatology ; : 121-127, 2015.
Article Dans Anglais | WPRIM | ID: wpr-156733

Résumé

BACKGROUND: Psoriasis is an autoimmune disease that is caused by a shift in the Th1/Th2 balance toward Th1-dominant immunity. It has been established as an effective treatment to counteract psoriasis by subcutaneous injection of recombinant interleukin (IL)-4, and IL-4 gene therapy by topical transdermal penetration has shown its antipsoriatic effect in mice. Retinoic acid (RA) and dimethylsulfoxide can increase the efficiency of gene transfection in the topical transdermal delivery system. OBJECTIVE: We investigated whether RA could improve anti-psoriasis efficiency using IL-4 expression plasmid pORF-mIL-4 (pIL-4) via transdermal delivery system in K14-vascular endothelial growth (K14-VEGF) factor transgenic mice. METHODS: After pretreatment with RA, plasmid pIL-4 in 10% dimethylsulfoxide was applied to the ear skin by topical transdermal penetration. Hematoxylin- eosin staining and immunohistochemistry were performed with ear samples to evaluate anti-psoriasis efficiency in mice. RESULTS: The psoriasis pathological features were relieved and psoriasis-associated factors were significantly reduced. CONCLUSION: Our results reveal that topical application of pIL-4 in dimethylsulfoxide by transdermal delivery with RA pretreatment can improve psoriasis significantly.


Sujets)
Animaux , Souris , Maladies auto-immunes , Diméthylsulfoxyde , Oreille , Éosine jaunâtre , Thérapie génétique , Immunohistochimie , Injections sous-cutanées , Interleukine-4 , Interleukines , Souris transgéniques , Plasmides , Psoriasis , Peau , Transfection , Trétinoïne
2.
Journal of Breast Cancer ; : 25-32, 2014.
Article Dans Anglais | WPRIM | ID: wpr-7630

Résumé

PURPOSE: The universal organic solvent dimethyl sulfoxide (DMSO) can be used as a differentiation inducer of many cancer cells and has been widely used as a solvent in laboratories. However, its effects on breast cancer cells are not well understood. The aim of this study is to investigate the effect and associated mechanisms of DMSO on mouse breast cancer. METHODS: We applied DMSO to observe the effect on tumors in a mouse breast cancer model. Tumor-associated macrophages (TAMs) were tested by flow cytometry. Ex vivo tumor microenvironment was imitated by 4T1 cultured cell conditioned medium. Enzyme-linked immunosorbent assays were performed to detect interleukin (IL)-10 and IL-12 expression in medium. To investigate the cytotoxicity of DMSO on TAMs, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed. RESULTS: We found that DMSO produced tumor retardation when injected into mouse peritoneal cavities in a certain concentration range (0.5-1.0 mg/g). Furthermore, as detected by flow cytometry, TAM subtypes were found to be transformed. We further imitated a tumor microenvironment in vitro by using 4T1 cultured cell conditioned medium. Similarly, by using low concentration DMSO (1.0%-2.0% v/v), TAMs were induced to polarize to the classically activated macrophage (M1-type) and inhibited from polarizing into the alternatively activated macrophage (M2-type) in the conditioned medium. IL-10 expression in tumors was reduced, while IL-12 was increased compared with the control. Furthermore, we reported that 2.0% (v/v) DMSO could lead to cytotoxicity in peritoneal macrophages after 48 hours in MTT assays. CONCLUSION: Our findings suggest that DMSO could exert antitumor effects in 4T1 cancer-bearing mice by reversing TAM orientation and polarization from M2- to M1-type TAMs. These data may provide novel insight into studying breast cancer immunotherapy.


Sujets)
Animaux , Souris , Tumeurs du sein , Région mammaire , Cellules cultivées , Milieux de culture conditionnés , Diméthylsulfoxyde , Test ELISA , Cytométrie en flux , Immunothérapie , Interleukine-10 , Interleukine-12 , Interleukines , Macrophages , Macrophages péritonéaux , Microenvironnement tumoral
3.
Acta Pharmaceutica Sinica ; (12): 322-328, 2014.
Article Dans Chinois | WPRIM | ID: wpr-245082

Résumé

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.


Sujets)
Animaux , Mâle , Souris , Anticorps , Métabolisme , Anticorps bispécifiques , Allergie et immunologie , Utilisations thérapeutiques , Réaction antigène-anticorps , Arthrite expérimentale , Métabolisme , Thérapeutique , Polyarthrite rhumatoïde , Métabolisme , Thérapeutique , Collagène de type II , Allergie et immunologie , Interleukine-17 , Métabolisme , Interleukine-1 bêta , Métabolisme , Interleukine-2 , Métabolisme , Rate , Métabolisme , Facteur de nécrose tumorale alpha , Métabolisme
4.
Chinese Journal of Cancer ; (12): 796-801, 2010.
Article Dans Anglais | WPRIM | ID: wpr-296352

Résumé

<p><b>BACKGROUND AND OBJECTIVE</b>iASPP, an inhibitory member of the apoptosis-stimulating proteins of p53 (ASPP) family, has been found to be up-regulated in various human tumor types. This study was to construct an efficient doxycycline-regulated, lentiviral vector-mediated knockdown system for iASPP that will allow for inducible down-regulation of iASPP gene expression and preliminary functional analysis.</p><p><b>METHODS</b>A pair of complementary oligos with hairpin structures targeting the iASPP gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced. The fragment containing the shRNA cassette was cloned to pLVCT-tTR-KRAB plasmid. The recombinant vectors were co-transfected with viral packaging mix into 293T cells, and viral supernatant was harvested to determine the titer. After treatment with or without doxycycline, HepG2 cells infected with virus were harvested and the expression of iASPP was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. Its effects on tumor growth were characterized using MTS assay, soft agar colony formation, and flow cytometry analysis.</p><p><b>RESULTS</b>The lentiviral vector expressing shRNA that targets to the oncogene iASPP was constructed successfully. HepG2 infected with the lentivirus expressing shRNA against iASPP inhibited the expression of iASPP in the presence of doxycycline, which resulted in the repression of tumor cell proliferation and anchorage-independent growth potential.</p><p><b>CONCLUSIONS</b>The lentiviral vector-mediated tet-on system demonstrates efficient and inducible knockdown of iASPP in hepatocellular carcinoma cells. iASPP gene may be involved in tumorigenesis and progression of human tumors.</p>


Sujets)
Humains , Apoptose , Carcinome hépatocellulaire , Génétique , Métabolisme , Anatomopathologie , Prolifération cellulaire , Régulation négative , Doxycycline , Pharmacologie , Vecteurs génétiques , Cellules HepG2 , Protéines et peptides de signalisation intracellulaire , Génétique , Métabolisme , Lentivirus , Génétique , Tumeurs du foie , Génétique , Métabolisme , Anatomopathologie , Plasmides , Interférence par ARN , Petit ARN interférent , Génétique , Protéines recombinantes , Génétique , Métabolisme , Protéines de répression , Génétique , Métabolisme , Transfection
5.
Biol. Res ; 42(1): 121-132, 2009. ilus
Article Dans Anglais | LILACS | ID: lil-519091

Résumé

Skeletal muscles have the potential to regenerate by activation of quiescent satellite cells, however, the molecular signature that governs satellite cells during muscle regeneration is not well defined. Myosin light chains (Myls) are sarcomere-related proteins as traditional regulator of muscle contraction. In this report, we studied the possible role of Myl in the proliferation of skeletal muscle-derived myoblasts. Compared to diaphragm-derived myoblasts, the extraocular muscle-derived myoblasts with lower levels of Myl proliferated faster, maintained a longer proliferation phase, and formed more final myotubes. It was found that blockading Myl with anti-Myl antibody or knockdown of Myll by siRNA targeted against Myll could enhance the myoblast proliferation and delay the differentiation of myoblasts. Our results suggested that Myl, likely Myll, can negatively affect myoblast proliferation by facilitating myoblast withdrawal from cell cycle and differentiation.


Sujets)
Animaux , Souris , Prolifération cellulaire , Muscle diaphragme/cytologie , Myoblastes/physiologie , Chaînes légères de myosine/physiologie , Muscles oculomoteurs/cytologie , Régénération/physiologie , Technique de Western , Immunohistochimie , RT-PCR
6.
Acta Pharmaceutica Sinica ; (12): 758-763, 2009.
Article Dans Chinois | WPRIM | ID: wpr-344110

Résumé

This investigation is to explore the feasibility of applying reverse docking method to the selectivity studies of protein kinase inhibitors. Firstly, a database that consists of 422 protein kinase structures was established through collecting the reported crystal structures or homology modeling. Then a reverse docking based method of protein kinase target screening was established, followed by the optimization of related parameters and scoring functions. Finally, seven typical selective kinase inhibitors were used to test the established method. The results show that the selective targets of these inhibitors have relatively high scoring function values (ranking in the first 35% of the tested kinase targets according to the scoring function values). This implies that the reverse docking method can be applied to the virtual screening of kinase targets and further to the selectivity studies of protein kinase inhibitors.


Sujets)
Épissage alternatif , Systèmes de délivrance de médicaments , Évaluation préclinique de médicament , Méthodes , Ciblage de gène , Modèles moléculaires , Liaison aux protéines , Inhibiteurs de protéines kinases , Chimie
7.
Experimental & Molecular Medicine ; : 617-628, 2008.
Article Dans Anglais | WPRIM | ID: wpr-59826

Résumé

Honokiol is an active compound purified from magnolia that has been shown to induce cell differentiation, apoptosis, and anti-angiogenesis effects, as well as an enhancement in tumor growth delay in combination with chemotherapeutic agents in several mouse xenograft models. Our goal was to investigate the radiosensitization effect of honokiol on lung carcinoma. The radiosensitization effect of liposomal honokiol in Lewis lung carcinoma cells (LL/2) was analyzed using an in vitro clonogenic survival assay. For an in vivo study, Lewis lung carcinoma-bearing C57BL/6 mice were treated with either liposomal honokiol at 25 mg/kg or 5 Gy of single tumor radiation, or a combination of both over 12 days of treatment. The tumor growth delay and the survival time were evaluated. In addition, histological analysis of tumor sections was performed to examine changes by detecting the microvessel density and apoptosis in tumor tissues. In the clonogenic survival assay, LL/2 cells treated with IC50 Lipo-HNK for 24 h showed a radiation enhancement ratio of 1.9. After 12 days of combination treatment, the tumor volume decreased 78% and produced an anti-tumor activity 1.3-fold greater than a predicted additive effect of honokiol and radiation alone. This combination treatment also caused an 8.7 day delay in tumor growth. The cell cycle distribution and histological analysis demonstrated that liposomal honokiol has an anti-tumor effect via inducing apoptosis and inhibiting angiogenesis. Liposomal honokiol can enhance tumor cell radiosensitivity in vitro and in vivo, indicating that radiotherapy combined with liposomal honokiol can lead to greater anti-tumor efficacy.


Sujets)
Animaux , Humains , Souris , Inhibiteurs de l'angiogenèse/administration et posologie , Apoptose , Dérivés du biphényle/administration et posologie , Carcinome pulmonaire de Lewis/traitement médicamenteux , Cycle cellulaire/effets des médicaments et des substances chimiques , Lignée cellulaire tumorale , Association thérapeutique , Lignanes/administration et posologie , Liposomes , Tumeurs du poumon/traitement médicamenteux , Magnolia/composition chimique , Transplantation tumorale , Néovascularisation pathologique/traitement médicamenteux , Radiotolérance , Transplantation hétérologue
8.
Chinese Medical Journal ; (24): 1433-1438, 2008.
Article Dans Anglais | WPRIM | ID: wpr-293985

Résumé

<p><b>BACKGROUND</b>Cyclin B1 (CLB1) is necessary for mitotic initiation in mammalian cells and plays important roles in cancer development. Therefore, a potential strategy in cancer therapy is to suppress the activity of CLB1 by delivering antisense constructs of CLB1 into tumor cells. In previous CLB1 studies, antisense constructs with a short half life were often used and these constructs might not persistently inhibit CLB1.</p><p><b>METHODS</b>We successfully created a recombinant plasmid encoding the full-length antisense cDNA of mouse cyclin B1 (AS-mCLB1) and transfected this construct to the murine Lewis lung carcinoma (LL/2) and CT-26 colon carcinoma (CT-26) cells. We isolated clones of LL/2 and CT-26 transfectants with stable expression of AS-mCLB1. Reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression of the mRNA and protein levels of CLB1. To further test the efficacy of this strategy in vivo, AS-mCLB1-expressing LL/2 and CT-26 transfectants were implanted into mice.</p><p><b>RESULTS</b>We found the expression of the mRNA and protein levels of CLB1 decrease in these transfectants. The inhibition of CLB1 caused prominent G1 arrest, abnormal morphology, retarded cell growth and an increase in apoptosis. In AS-mCLB1-expressing LL/2 and CT-26 transfectants implanted mice, tumorigenicity was effectively suppressed compared with the controls. In addition, the expression of AS-mCLB1 also significantly increases the survival duration of implanted animals.</p><p><b>CONCLUSION</b>AS-mCLB1 is likely to be useful in future cancer therapy, which may be associated with its ability to down-regulate the expression of CLB1 and then induce G1arrest and apoptosis in tumor cells.</p>


Sujets)
Animaux , Souris , Apoptose , Prolifération cellulaire , Survie cellulaire , Cycline B , Génétique , Cycline B1 , ADN antisens , Pharmacologie , ADN complémentaire , Pharmacologie , Phase G1 , Souris de lignée C57BL , Transplantation tumorale , Tumeurs expérimentales , Anatomopathologie , Thérapeutique
9.
Journal of Experimental Hematology ; (6): 768-771, 2008.
Article Dans Chinois | WPRIM | ID: wpr-267892

Résumé

The purpose of this study was to investigate the effects of chloroquine diphosphate on the proliferation and apoptosis of human leukemic K562 cells, and to elucidate its possible mechanism of activity. The inhibitory effect of chloroquine diphosphate with different concentrations on K562 cell proliferation was detected by MTT method. Apoptosis was measured by flow cytometry (FCM); morphological analysis of apoptosis was performed after staining with propidium iodide (PI) under fluorescence microscope; cell apoptosis was assessed by the DNA ladder shown agarose gel electrophoresis. After treatment with chloroquine diphosphate, K562 cells were stained by Rhodamine 123 to detect changes in mitochondrial transmembrane potential (DeltaPsim) by FCM. The results showed that the cell viability decreased in dose-dependent manner, following chloroquine diphosphate treatment at different concentrations (1.5625, 3.125, 6.25, 12.5, 25, 50 and 100 micromol/L) for 24, 48 and 72 hours. By FCM analysis, the significant increases of sub-G(1) were observed. DNA ladder was detected and apoptotic nuclei were observed. DeltaPsim decreased in K562 cells after chloroquine diphosphate treatment. It is concluded that the chloroquine diphosphate can inhibit the proliferation of K562 cells and induce cell apoptosis, which may relate to down-regulation of mitochondrial transmembrane potential (DeltaPsim).


Sujets)
Humains , Antinéoplasiques , Pharmacologie , Apoptose , Prolifération cellulaire , Chloroquine , Pharmacologie , Relation dose-effet des médicaments , Régulation négative , Cellules K562 , Potentiels de membrane , Mitochondries
10.
Chinese Journal of Biotechnology ; (12): 423-428, 2007.
Article Dans Chinois | WPRIM | ID: wpr-328011

Résumé

It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.


Sujets)
Humains , Protéines régulatrices de l'apoptose , Génétique , Métabolisme , Séquence nucléotidique , Technique de Western , Lignée cellulaire , Clonage moléculaire , Expression des gènes , Gènes myc , Génétique , Vecteurs génétiques , Génétique , Protéines à fluorescence verte , Génétique , Métabolisme , Microscopie de fluorescence , Données de séquences moléculaires , Protéines de fusion recombinantes , Génétique , Métabolisme , Transfection
11.
Chinese Journal of Medical Genetics ; (6): 63-66, 2006.
Article Dans Chinois | WPRIM | ID: wpr-263851

Résumé

Angiopoietins(ANGPT) and their endothelial cell-specific tyrosine kinase receptors TEK are the major regulators of blood vessels angiogenesis under physiological and pathologic conditions. ANGPT1 is essentially involved in maturation, stabilization, and remodeling of blood vessels through inducing TEK autophosphorylation, promoting endothelial cell migration and survival. Instead, ANGPT2 appears to act as a natural antagonist of ANGPT1, it can activate vascular remodeling with the presence of vascular endothelial growth factor(VEGF) or regress frank blood vessels under the absence of VEGF. High expression of angiopoietins and TEK is often detected in tumor tissues. Many studies showed that disrupting the ANGPT/TEK receptor pathway could inhibit the growth of a number of murine tumors and human tumors. Thus, it is possible that inhibitors targeting the ANGPT/TEK pathway will have broad clinical utility to treatment of cancer.


Sujets)
Humains , Angiopoïétine-1 , Physiologie , Angiopoïétine-2 , Physiologie , Angiopoïétines , Physiologie , Néovascularisation physiologique , Physiologie , Récepteur TIE-2 , Physiologie
12.
West China Journal of Stomatology ; (6): 534-536, 2005.
Article Dans Chinois | WPRIM | ID: wpr-289022

Résumé

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector expressing hCD40L gene and explore it in the use of anti-tumor gene therapy.</p><p><b>METHODS</b>1,900 bp gene fragment was obtained form plasmid pORF-hCD40L by Xho I/Swa I cutting and then cloned directionally into the pShuttle plasmid, finally, the resultant plasmid was digested by restriction endonnuclease PmeI and subsequently cotransformtion into BJ5183 cells with the adenoviral backbone pAdEasy-1 to obtain the homologous recombinant and then the recombinant was packaged in the 293 cells. Some methods such as PCR and endonulease digestion were employed to identify the recombinant adenovirus.</p><p><b>RESULTS</b>The evidences of endonulease digestion and PCR analysis confirmed that recombinant hCD40L gene was correctly inserted into adenovirus vector.</p><p><b>CONCLUSION</b>The adenoviral vector which expressed hCD40L gene was constructed. It provides an experimental basis for studies on it expression in the mammalian cells and in tumor gene therapy.</p>


Sujets)
Animaux , Humains , Adenoviridae , Adénovirus humains , Ligand de CD40 , Thérapie génétique , Vecteurs génétiques , Plasmides , Réaction de polymérisation en chaîne
13.
Chinese Journal of Medical Genetics ; (6): 101-105, 2004.
Article Dans Chinois | WPRIM | ID: wpr-329389

Résumé

<p><b>OBJECTIVE</b>To study the prokaryotic expression of extracellular ligand binding domains of chick tie-2, the purification, refolding conditions of the recombinant protein, and its anti-angiogeneic effect.</p><p><b>METHODS</b>A DNA fragment encoding extracellular ligand binding domains of chick tie-2 was obtained by PCR amplification using a previous constructed plasmid as a template. The amplified fragment was then inserted into prokaryotic expression vector pQE30, and was expressed in E.Coli XL-1 blue by adding isopropyl-beta-D-thiogalactoside(IPTG). The recombinant protein in inclusion bodies was purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography under denatured conditions. Then the refolding of the purified protein was performed with gradient dialysis. The target protein was injected s.c. into mouse, and the antibody was detected by ELISA and Western blot analysis. The antibody was purified from the antiserum and then incubated with human umbilical endothelial vein cell (HUEVC) to find its anti-angiogenesis in vitro by using propidium iodide(PI) dying through FACS. Alginate encapsulated tumor cell assays were performed and micro-vessel density was determined by counting per high power field in the sections stained with an antibody reactive to CD31 to test its inhibition of angiogenesis.</p><p><b>RESULTS</b>The recombinant protein was highly expressed in E.Coli XL-1 blue, and the antibody produced in mouse could specifically recognize the recombinant protein. The purified antibody could induce apoptosis of HUEVC in vitro. The anti-angiogenic effect of the antibody could also be found in alginate-encapsulate tumor cell assay and by counting micro-vessel density.</p><p><b>CONCLUSION</b>The protein of extracellular ligand binding domains of chick tie-2 can be expressed at high level in the prokaryotic expression system, and the expressed protein can induce immune response in mouse. Furthermore, the antibody can induce the anti-angiogenic effect.</p>


Sujets)
Animaux , Souris , Inhibiteurs de l'angiogenèse , Pharmacologie , Sites de fixation , Technique de Western , Poulets , Test ELISA , Escherichia coli , Génétique , Souris de lignée BALB C , Souris de lignée C57BL , Antigènes CD31 , Récepteur TIE-2 , Chimie , Métabolisme , Protéines recombinantes , Pharmacologie
14.
Chinese Journal of Medical Genetics ; (6): 557-561, 2004.
Article Dans Chinois | WPRIM | ID: wpr-321196

Résumé

<p><b>OBJECTIVE</b>The growth and metastasis of solid tumors are dependent on angiogenesis. Endostatin, the C-terminal proteolytic fragment of collagen XVIII, is a potent endogenous angiogenesis inhibitor. The authors designed a topical antiangiogenic gene therapy with recombinant human endostatin adenovirus (Ad-hEndo) and assessed its effects on the inhibition of angiogenesis in vitro, and tumor growth and metastasis in vivo.</p><p><b>METHODS</b>Malignant cells (A549) were infected with Ad-hEndo. The expression of recombinant protein and the inhibition of cultured human umbilical vein endothelial were investigated. Immunodeficient A549 nude mice were treated with intratumoral injection of Ad-hEndo, the empty vector Ad-control or saline (NS). The dose-response, side effects, and serum concentration of endostatin were observed.</p><p><b>RESULTS</b>Recombinant endostatin protein was detected in the infected tumor cells with different MOI Ad-hEndo and its inhibitory effect on endothelial cells growth was shown. In animal study, the volume of tumor and the number of pulmonary metastatic lesions in the Ad-hEndo treatment group were significantly smaller than those in the control groups (P<0.05).</p><p><b>CONCLUSION</b>The present findings provide evidence of the anti-tumor effects of the endostatin and may be important for the further use of it in topical antiangiogenic gene therapy of cancer.</p>


Sujets)
Animaux , Humains , Souris , Adenoviridae , Génétique , Lignée cellulaire tumorale , Prolifération cellulaire , Cellules cultivées , Modèles animaux de maladie humaine , Endostatines , Génétique , Utilisations thérapeutiques , Cellules endothéliales , Thérapie génétique , Vecteurs génétiques , Rein , Biologie cellulaire , Métabolisme , Souris nude , Métastase tumorale , Transplantation tumorale , Tumeurs expérimentales , Répartition aléatoire , Protéines recombinantes , Génétique , Utilisations thérapeutiques , Transfection , Veines ombilicales , Biologie cellulaire , Tests d'activité antitumorale sur modèle de xénogreffe
15.
West China Journal of Stomatology ; (6): 145-148, 2004.
Article Dans Chinois | WPRIM | ID: wpr-319034

Résumé

<p><b>OBJECTIVE</b>The purpose of this study was to clone the soluble form of the mouse BlyS (msBlyS) and insert it into a eukaryotic expression vector pSecTag2B in order to further elucidat the antitumor activity induced by msBlyS expressed by the recombined plasmid pSecTag2B-msBlyS.</p><p><b>METHODS</b>Full length cDNA of mouse soluble BlyS (msBlyS) was amplified by reverse transcription-PCR from total RNA of mouse spleen. The PCR product was ligated directly with linearized vector pCR2.1 supplied in the TA cloning kit. The recombined plasmid pCR2.1-msBlyS which was selected and identified using blue-white screening method and restriction map analysis and the purified original plasmid pSecTag2B were both cut by HindIII and EcoR I. The digested fragments were extracted and purified from low-melting temperature agarose and ligated by T4DNA ligase. The recombined plasmid pSecTag2B-msBlyS were isolated and identified by restricted endonuclease cutting and Sanger dideoxy DNA sequencing.</p><p><b>RESULTS</b>The sequencing data indicated that inserted msBlyS gene had correct DNA sequence and orientation.</p><p><b>CONCLUSION</b>Eukaryotic expression vector pSecTag2B. Expressing mouse BlyS have successfully been cloned. This will provide us an opportunity to do further research work on BlyS.</p>


Sujets)
Animaux , Souris , Facteur d'activation des lymphocytes B , Clonage moléculaire , Déterminants antigéniques des lymphocytes B , Génétique , Cellules eucaryotes , Métabolisme , Vecteurs génétiques , Protéines membranaires , Génétique , Souris de lignée BALB C , Plasmides , Génétique , Réaction de polymérisation en chaîne , Récepteurs aux facteurs de nécrose tumorale , Génétique , Recombinaison génétique , Analyse de séquence d'ADN , Rate , Biologie cellulaire , Allergie et immunologie , Facteur de nécrose tumorale alpha , Génétique
16.
Chinese Journal of Oncology ; (12): 119-121, 2004.
Article Dans Chinois | WPRIM | ID: wpr-271052

Résumé

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of capecitabine as first-line therapy in patients with advanced and recurrent colorectal cancer.</p><p><b>METHODS</b>From December 2000 to November 2001, sixty patients with advanced and recurrent colorectal cancer received first-line capecitabine treatment given at a dose of 1250 mg/m(2) twice daily, on days 1 - 14 every 21 days. At least 2 cycles were administered.</p><p><b>RESULTS</b>The overall response rate was 23.3% with 14 PR, 24 SD (40.0%) and 15 PD. The median survival time was 14.7 months. The survival rate was 63.9% at 12-months and 33.4% at 24-months. Grade III-IV adverse effects were diarrhea in 4 patients (6.6%), anemia in 2 (3.3%) and hand-foot syndrome (HFS) in 1 (1.7%); Grade I-II adverse effects were hyperpigmentation in 20 (33.3%), HFS in 18 (30.0%) and diarrhea in 10 (16.7%).</p><p><b>CONCLUSION</b>Capecitabine is an efficacious and better-tolerated alternative treatment for the patients with advanced and recurrent colorectal cancer.</p>


Sujets)
Adulte , Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Antimétabolites antinéoplasiques , Utilisations thérapeutiques , Capécitabine , Tumeurs colorectales , Traitement médicamenteux , Mortalité , Désoxycytidine , Utilisations thérapeutiques , Fluorouracil , Taux de survie
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