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1.
Chinese Journal of Experimental and Clinical Virology ; (6): 347-351, 2018.
Artigo em Chinês | WPRIM | ID: wpr-806318

RESUMO

Objective@#To investigate the cellular and humoral immune responses induced by combined immunization with the fusion protein of human papillomavirus type 18 (HPV18) and the recombinant vaccinia virus.@*Methods@#Purified HPV18L231-600E7E6 fusion protein, expressed by prokaryotic expression system, were immunized in combination with the recombinant vaccinia virus vaccine expressing HPV18E7E6 fusion protein (rVV18E7E6) by using various prime-boost regiments in C57BL/6 mice. Cellular and humoral immune responses were analyzed by enzyme-linked immunospot assay (ELISPOT), enzyme-linked immunosorbent assay (ELISA), and pseudovirus neutralization assay.@*Results@#Higher levels of cellular immune responses were induced in mice primed with the HPV18L231-600E7E6 fusion protein/adjuvant CpG and boosted with the recombinant vaccinia virus rVV18E7E6, than in other immunized mice. Higher binding antibody level was induced, and low level neutralizing antibody against pseudovirus was detected simultaneously.@*Conclusions@#Priming with HPV18L231-600E7E6 fusion protein/CpG and boosting with the recombinant vaccinia virus rVV18E7E6 could induce higher cellular and humoral immune response in immunized mice, which might be taken as vaccine candidate for treatment of HPV18 chronic infection and postoperative adjuvant treatment for cervical cancer.

2.
Chinese Journal of Virology ; (6): 432-436, 2013.
Artigo em Chinês | WPRIM | ID: wpr-339932

RESUMO

In order to facilitate the detection of newly emerging or rare viral infectious diseases, a negative-strand RNA virus-severe fever with thrombocytopenia syndrome bunyavirus, and a positive-strand RNA virus-dengue virus, were used to investigate RNA viral genome unspecific amplification by multiple displacement amplification technique from clinical samples. Series of 10-fold diluted purified viral RNA were utilized as analog samples with different pathogen loads, after a series of reactions were sequentially processed, single-strand cDNA, double-strand cDNA, double-strand cDNA treated with ligation without or with supplemental RNA were generated, then a Phi29 DNA polymerase depended isothermal amplification was employed, and finally the target gene copies were detected by real time PCR assays to evaluate the amplification efficiencies of various methods. The results showed that multiple displacement amplification effects of single-strand or double-strand cDNA templates were limited, while the fold increases of double-strand cDNA templates treated with ligation could be up to 6 X 10(3), even 2 X 10(5) when supplemental RNA existed, and better results were obtained when viral RNA loads were lower. A RNA viral genome amplification system using multiple displacement amplification technique was established in this study and effective amplification of RNA viral genome with low load was achieved, which could provide a tool to synthesize adequate viral genome for multiplex pathogens detection.


Assuntos
Humanos , Infecções por Bunyaviridae , Diagnóstico , Virologia , Linhagem Celular , DNA Ligases , Metabolismo , DNA Complementar , Genética , DNA Polimerase Dirigida por DNA , Metabolismo , Dengue , Diagnóstico , Virologia , Vírus da Dengue , Genética , Genoma Viral , Genética , Phlebovirus , Genética , RNA Viral , Genética , Padrões de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Métodos , Carga Viral
3.
Chinese Journal of Virology ; (6): 349-356, 2013.
Artigo em Chinês | WPRIM | ID: wpr-356680

RESUMO

Viral hemorrhagic fevers (VHFs) refer to a group of acute infections with high case fatality rates that are caused by four distinct families of RNA viruses belonging to the families Bunyaviridae, Flaviviridae, Filoviridae and Arenaviridae, the main clinical symptoms of these diseases are accompanied by fever and bleeding. For the reason that these infections have similar primary clinical symptoms, it is difficult to diagnose and distinguish them; rapid and reliable laboratory diagnostic tests are required in suspected cases for epidemiological investigation and controlling the spread of VHFs. This review addresses the laboratory diagnostics of VHFs, covering etiological classification and different diagnostic techniques, such as virus isolation, nucleic acid detection, as well as antigen and antibody assays. Prospects for novel diagnostic tools are also discussed.


Assuntos
Humanos , Técnicas de Laboratório Clínico , Métodos , Febres Hemorrágicas Virais , Diagnóstico , Alergia e Imunologia , Virologia , Vírus de RNA , Genética , Alergia e Imunologia
4.
Chinese Journal of Microbiology and Immunology ; (12): 276-280, 2012.
Artigo em Chinês | WPRIM | ID: wpr-428757

RESUMO

ObjectiveTo construct one recombinant adenovirus AdE7E6 expressing HPV16 E6 and E7 fusion protein as candidate for HPV16 therapeutic vaccine.MethodsThe codon-optimized E6 and E7 gene,were fused to create one open reading frame,then inserted into adenovirus vector pCD316.A strain of recombinant adenovirus was constructed through homologous recombinant in 293 cells,and identified by PCR and Western blot.Finally,it was employed to study it's immunogenicity and the activity of the tumor growth regression.ResultsThe PCR result showed that E6E7 fusion gene had been integrated in recombinant Ad5 DNA.Western blot test confirmed that the E6E7 fusion protein was highly expressed in 293 cells infected with Ad5E7E6 recombinant adenovirus.The recombinant adenovirus elicited significant E7 specific CD8+ T lymphocyte response in vaccinated mice.These responses could completely prevent the TG-1 tumor cell bearing mice treated with AdE7E6 from developing into tumor.ConclusionThese results suggested that rAd5E7E6 could be a potent vaccine candidate for the treatment of HPV16-associated tumors and their precancerous transformations.

5.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (24): 980-986, 2010.
Artigo em Chinês | WPRIM | ID: wpr-747472

RESUMO

OBJECTIVE@#To study the self-assessment, acoustic analysis, laryngoscopy and its relationship for patients with voice disorders before and after surgery.@*METHODS@#Fifty patients with voice disorders were undergone self-assessment, acoustic analysis, and laryngoscopy before and after voice surgery. Self-assessment were done by voice handicap index (VHI) Scale Chinese version, including functional (F), physiological (P), emotion (E) and its sum denoted as T. Acoustic analysis were made for patient samples by Dr. Speech voice analysis software and jitter (J), shimmer (S), normalized noise energy (NNE) were selected as three parameters. Laryngoscopic examination were used to record the closure of vocal cord morphologically (C).@*RESULT@#In addition to E, F, P and T(VH) of VHI scale had a good correlation. In acoustic analysis J, S and NNE had a good correlation between them. F, P and T(VH) of VHI scale and acoustic analysis parameters J, S and NNE had a good correlation. Closed degree C and the VHI scale F, P and T(VH) as well as acoustic analysis parameters J, S, NNE had a good correlation. All the above data use were analyzed by Pearson correlation test.@*CONCLUSION@#VHI scale Chinese version make the patient's subjective feelings as the center, thus it has some limitations for the impact of East-West cultural differences, age, educational level and other factors. Acoustic analysis can show a detailed objective aspect of the patient's voice quality and evaluate the result of surgical treatment. Laryngoscopy provide an excellent morphological evidence. Consistency of three methods can do a comprehensive assessment for the voice disease.


Assuntos
Adulto , Idoso , Humanos , Pessoa de Meia-Idade , Período Intraoperatório , Laringoscopia , Autoavaliação (Psicologia) , Acústica da Fala , Prega Vocal , Distúrbios da Voz , Diagnóstico , Cirurgia Geral , Qualidade da Voz
6.
Chinese Journal of Experimental and Clinical Virology ; (6): 171-174, 2010.
Artigo em Chinês | WPRIM | ID: wpr-316933

RESUMO

<p><b>OBJECTIVE</b>To express HPV6bL2deltaN360E7E6 fusion protein in E. coli and preliminarily evaluate its immune effect.</p><p><b>METHODS</b>Three HPV6b gene fragments, which were L2(1-360 bp), E7 and E6, were fused by overlapping PCR, then were inserted into a prokaryotic expression vector and expressed in E. coli. C57BL/6 mice were immunized with purified fusion protein plus Al (OH)3 and/or CpG adjuvants through intramuscular route, the cellular and humoral immune responses were detected by IFN-gamma ELISPOT and ELISA respectively.</p><p><b>RESULTS</b>Protein plus CpG adjuvant could induce the strongest cellular immune response to E7 and E6, high antibody titer against L2 could be detected in all immunized groups but there were no significant difference among these groups.</p><p><b>CONCLUSIONS</b>HPV6bL2deltaN360E7E6 gene was successfully cloned into pQE30 vector and expressed in E. coli, the fusion protein was also purified and proved that could induce strong cellular and humoral immune responses with appropriate adjuvant in C57BL/ 6 mice and could be used for future research.</p>


Assuntos
Animais , Camundongos , Adjuvantes Imunológicos , Condiloma Acuminado , Genética , Ensaio de Imunoadsorção Enzimática , ELISPOT , Métodos , Escherichia coli , Genética , Expressão Gênica , Vetores Genéticos , Imunidade Humoral , Imunização , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase , Proteínas Recombinantes de Fusão , Genética , Metabolismo , Farmacologia , Proteínas Recombinantes , Química , Genética
7.
Chinese Journal of Experimental and Clinical Virology ; (6): 406-408, 2010.
Artigo em Chinês | WPRIM | ID: wpr-316888

RESUMO

<p><b>OBJECTIVE</b>To identify specific T lymphocyte epitope on E6 protein of human papillomavirus type 18 in mice.</p><p><b>METHODS</b>Infection with one recombinant vaccinia virus rVVJ18 E7, E6 respectively in C57 BL/6 and BALB/c mice, specific cellular immune responses were detected by ELISPOT or intracellular cytokine stainings by using a series of overlapping synthetic peptides covering full length of the amino acid sequence of E6 and E7 proteins or various truncated peptides.</p><p><b>RESULTS</b>The rVVJ18 E7, E6 generated significant E6 specific T-cell immune responses in vaccinated mice. Mapping of the epitope of E6 revealed that the peptides E6(67-75 ( KCIDFYSRI) and E6(60-68) (IPHAAGHKC) presented respectively by C57BL/6 and BALB/c mice were the optimal peptides to activate E6-specific CD8+ T lymphocytes. However no positive cellular immune responses stimulated with various E7 peptides were detected by ELISPOT in immunized mice.</p><p><b>CONCLUSIONS</b>Two specific T lymphocyte epitopes were identified on E6 protein in C57BL/6 and BALB/c mice, which will provide the basis to evaluate cellular immune response elicited by HPV18 E6 protein based vaccine.</p>


Assuntos
Animais , Humanos , Camundongos , Linfócitos T CD8-Positivos , Alergia e Imunologia , Proteínas de Ligação a DNA , Alergia e Imunologia , Epitopos de Linfócito T , Alergia e Imunologia , Papillomavirus Humano 18 , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Proteínas Oncogênicas Virais , Alergia e Imunologia , Vacinas contra Papillomavirus , Alergia e Imunologia
8.
Acta Physiologica Sinica ; (6): 210-214, 2007.
Artigo em Chinês | WPRIM | ID: wpr-258668

RESUMO

The purpose of the present study was to explore the expression changes of intermedin/adrenomedullin 2 (IMD/ADM2), a novel small molecular bioactive peptide, and its receptors, calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMP1, RAMP2, RAMP3) in the right ventricle of rats with chronic hypoxia-induced pulmonary hypertension. Twenty male Sprague-Dawley rats were randomly divided into 4-week hypoxia group and normal control group (each n=10). The rats in hypoxia group were placed in an isobaric hypoxic chamber, in which O(2) content was maintained at 9%-11% by delivering N(2), and CO(2) content was maintained at <3% for 4 weeks (8 h/d, 6 d/week). The rats in the control group were housed in room air. The protein levels of IMD/ADM2 and adrenomedullin (ADM) in blood plasma and right ventricular tissue were measured by radioimmunoassay. The mRNA expressions of IMD/ADM2, ADM and their receptors CRLR, RAMP1, RAMP2, RAMP3 in right ventricular tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the ratio of right ventricle weight to left ventricle plus septum weight [RV/(LV+S)] and mean pulmonary arterial pressure (mPAP) were higher in hypoxia group than those in the control group (all P<0.01), suggesting that the rat model of pulmonary hypertension was successfully established. However, the mean carotid arterial pressure (mCAP) between the two groups had no significant difference. Compared with that in the control group, ADM contents in plasma and right ventricular tissue in hypoxia group increased by 1.26 and 1.68 folds (all P<0.01), respectively. Likewise, IMD/ADM2 contents in blood plasma and right ventricular tissue in hypoxia group increased by 0.90 and 1.19 folds (P<0.01), respectively, compared with that in the control group. The data of RT-PCR showed that mRNA levels of ADM, IMD/ADM2 and RAMP2 in hypoxia group increased by 155.1% (P<0.01), 80.9% (P<0.01) and 52.9% (P<0.05), respectively, compared with those in the control group. There were no significant differences in mRNA expressions of CRLR, RAMP1 and RAMP3 between the two groups (all P>0.05). Taken together, the results show that the level of IMD/ADM2 increases in the rats with chronic hypoxia-induced pulmonary hypertension.


Assuntos
Animais , Masculino , Ratos , Adrenomedulina , Metabolismo , Proteína Semelhante a Receptor de Calcitonina , Metabolismo , Ventrículos do Coração , Metabolismo , Hipertensão Pulmonar , Metabolismo , Hipóxia , Neuropeptídeos , Metabolismo , Ratos Sprague-Dawley , Proteínas Modificadoras da Atividade de Receptores , Metabolismo
9.
Chinese Journal of Applied Physiology ; (6): 467-471, 2007.
Artigo em Chinês | WPRIM | ID: wpr-310832

RESUMO

<p><b>AIM</b>To investigate the changes and probable roles of adrenomedullin2/intermedin (AIDM2/IMD), a novel micromolecular bioactive peptide, in the lungs of rats with chronic hypoxic pulmonary hypertension.</p><p><b>METHODS</b>Twenty male SD rats were randomly divided into normal control group (NC) and normobaric hypoxia group (4H). The protein levels of ADM and ADM2/IMD) in the plasma and lung were measured by radioimmunoassay and immunohistochemistry. The mRNA expressions of ADM, ADM2/IMD and their receptors C (RLR, RAMP1, RAMP2 and RAMP3 in the lung tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The rat model of chronic pulmonary hypertension was confirmed by the increased mean pulmonary arterial pressure (mPAP) and weight ratio of right ventricle to left ventricle plus septum [RV/(LV + S)] in 4H group compared to NC group. (2) The concentrations of ADM in the plasma and lung homogenate of 4H group were 2.3 and 3.2 folds of NC group, respectively (all P < 0.01). The levels of ADM2/IMD were higher 89.6% and 45.0% in the plasma and lung homogenate of 4H group than those of NC group (respectively, P < 0.01, P < 0.05). (3) The mRNA expressions of ADM2/IMD and ADM in the lung of 4H group were up-regulated (respectively, P < 0.01, P < 0.05 vs. NC group). The expressions of CRLR and RAMP1 mRNAs were down-regulated (all P < 0.01 vs. NC group), while the levels of RAMP2 and RAMP3 mRNAs were no significant difference between the two groups. (4) The strong ADM2/IMD immunostaining was detected in the endothelial and adventitial cells of the rat pulmonary arteriole.</p><p><b>CONCLUSION</b>ADM2/IMD, like its paralog ADM, might be closely related to the chronic hypoxic pulmonary hypertension in rats. The disorders of the gene expression and/or the synthesis and metabolism of ADM2/IMD and its receptor CRLR/RAMP1 possibly take part in the pathogenesis of chronic hypoxic pulmonary hypertension in rats.</p>


Assuntos
Animais , Masculino , Ratos , Adrenomedulina , Metabolismo , Hipertensão Pulmonar , Metabolismo , Hipóxia , Metabolismo , Pulmão , Metabolismo , Neuropeptídeos , Metabolismo , Ratos Sprague-Dawley
10.
Chinese Medical Journal ; (24): 37-41, 2004.
Artigo em Inglês | WPRIM | ID: wpr-235837

RESUMO

<p><b>BACKGROUND</b>Human urotensin II (UII) is the most potent mammalian vasoconstrictor identified so far. Our previous study showed that UII is a potent mitogen of airway smooth muscle cells (ASMC) inducing ASMC proliferation in a dose-dependent manner. The signal transduction pathway of UII mitogenic effect remains to be clarified. This study was conducted to investigate the signal transduction pathway in the proliferation of ASMC induced by UII.</p><p><b>METHODS</b>In primary cultures of rat ASMCs, activities of protein kinase C (PKC), mitogen-activated protein kinase (MAPK) and calcineurin (CaN) induced by UII were measured. The effect of CaN on PKC and MAPK was studied by adding cyclosporin A (CsA), a specific inhibitor of CaN. Using H7 and PD98059, inhibitors of PKC and MAPK, respectively, to study the effect of PKC and MAPK on CaN. The cytosolic free calcium concentration induced by UII was measured using Fura-2/AM.</p><p><b>RESULTS</b>UII 10(-7) mol/L stimulated ASMC PKC and MAPK activities by 44% and 24% (P < 0.01), respectively, after incubating for 20 minutes. It increased CaN activity in a time-dependent manner, being 1.68 times as that of control for 24 hours (P < 0.01). It promoted the cytosolic free calcium concentration increase of 18% (P < 0.01). CsA 10(-6) mol/L and H7 50 micromol/L inhibited UII-stimulated CaN activity by 45% (P < 0.01) and 21% (P < 0.05), respectively, while PD98059 50 micromol/L had no effect on CaN activity (P > 0.05). CsA 10(-6) mol/L inhibited UII-stimulated PKC activity by 14% (P < 0.05), while having no effect on MAPK activity (P > 0.05).</p><p><b>CONCLUSIONS</b>UII increases cytosolic free calcium concentration and activates PKC, MAPK and CaN. The signal transduction pathway between PKC and CaN has cross-talk.</p>


Assuntos
Animais , Ratos , Calcineurina , Metabolismo , Células Cultivadas , Ativação Enzimática , Proteínas Quinases Ativadas por Mitógeno , Metabolismo , Mitógenos , Farmacologia , Miócitos de Músculo Liso , Biologia Celular , Proteína Quinase C , Metabolismo , Transdução de Sinais , Fisiologia , Traqueia , Biologia Celular , Urotensinas , Farmacologia
11.
Acta Physiologica Sinica ; (6): 260-264, 2003.
Artigo em Inglês | WPRIM | ID: wpr-290975

RESUMO

In this study, we observed the levels of adrenomedullin (ADM) and proadrenomedullin N-terminal 20 peptide (PAMP) in myocardium and aorta of spontaneously hypertensive rats (SHRs) in comparison with Wistar-kyoto (WKY) rats. Contents of ADM and PAMP were measured by radioimmunoassay (RIA) in plasma, myocardium and aorta. The amount of Pro-ADM mRNA of myocardium and aorta was determined by competitive quantitative reverse transcription polymerase chain reaction (RT-PCR). In SHRs the amounts of Pro-ADM mRNA of myocardium and aorta were 66.7% (P<0.01) and 73% (P<0.01) higher than those in WKY rat, respectively. In SHRs, the levels of ADM in plasma, myocardium and aorta were 29%, 76.7% and 79% (all P<0.01) higher than those in WKY rats, respectively. The level of PAMP in SHRs was increased by 42.5% in plasma (P<0.01), 47.2% in myocardium (P<0.0.1) and 27.3% in aorta (P<0.05) compared to WKY rats, respectively. In addition, the ratio of ADM content to PAMP content in SHRs group was increased compared with that in WKY group (2.0+/-0.25 vs 1.64+/-0.3 and 2.2+/-0.18 vs 1.56+/-0.28, in myocardium and aorta, respectively, P<0.01). These results suggest that ProADM gene expression is up-regulated and the increase in ADM and PAMP is different in SHRs. The significance of inconsistency of increase in ADM and PAMP in SHRs needs to be further investigated.


Assuntos
Animais , Feminino , Masculino , Ratos , Adrenomedulina , Genética , Metabolismo , Aorta , Metabolismo , Miocárdio , Metabolismo , RNA Mensageiro , Genética , Metabolismo , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Regulação para Cima
12.
Chinese Journal of Applied Physiology ; (6): 132-136, 2003.
Artigo em Chinês | WPRIM | ID: wpr-339660

RESUMO

<p><b>AIM</b>To investigate the alterations of taurine transport, and taurine transporter (TAUT) mRNA by hyperglycemia in cultured rat cardiomyocytes.</p><p><b>METHODS</b>3H-taurine measured the amount of taurine uptake. TAUT mRNA consents were measured using quantitative RT-PCR.</p><p><b>RESULTS</b>The cellular uptake amounts of taurine in seven groups increased with incubation time, and near to be saturated after 5 min. The uptake amount of 10, 20, and 30 mmol/L glucose groups was obviously lower than that of the control group (P < 0.05 or P < 0.01). In 30 mmmol/L glucose, taurine release obviously was decreased, as compared with that of the control. Exposure of cells to 10, 20, and 30 mmmol/L glucose decreased taurine uptake in a concentration-dependent fashion. Exposure to hyperglycemia did not affect the Km of the TAUT, but the apparent Vmax were significantly decreased (P < 0.05). In 20 and 30 mmmol/L groups, TAUT mRNA contents of myocardial cells were significantly reduced, as compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>The data suggests that there are dysfunction of taurine uptake and downregulation of TAUT gene expression by glucose in cultured rat cardiomyocytes.</p>


Assuntos
Animais , Ratos , Células Cultivadas , Glucose , Farmacologia , Hiperglicemia , Metabolismo , Glicoproteínas de Membrana , Genética , Metabolismo , Proteínas de Membrana Transportadoras , Genética , Metabolismo , Miócitos Cardíacos , Metabolismo , RNA Mensageiro , Genética , Ratos Sprague-Dawley , Taurina , Metabolismo
13.
Acta Academiae Medicinae Sinicae ; (6): 359-363, 2002.
Artigo em Chinês | WPRIM | ID: wpr-278165

RESUMO

<p><b>OBJECTIVE</b>To observe the alterations of taurine transport, taurine transporter (TAUT) and cysteine sulfinate decarboxylase (CSD) mRNA in the calcification of myocardial cells in vitro.</p><p><b>METHODS</b>3H-taurine measured the amount of taurine uptake. TAUT and CSD mRNA consents were measured using competitive quantitative RT-PCR in cultured and calcified myocardial cells.</p><p><b>RESULTS</b>In calcification of myocardial cells, taurine concentration was decreased by 27% (P < 0.05), taurine uptake was markedly reduced, Vmax reduced by 39% (P < 0.01), there were no statistical significance of Km values between the two groups. TAUT mRNA decreased by 45% (P < 0.01), but CSD mRNA increased by 25% (P < 0.05).</p><p><b>CONCLUSIONS</b>The data suggest that there were impediment of taurine transport in calcification of myocardial cells, as TAUT mRNA level was decreased, but CSD mRNA concentration was improved.</p>


Assuntos
Animais , Ratos , Transporte Biológico , Calcinose , Metabolismo , Patologia , Cálcio , Metabolismo , Carboxiliases , Metabolismo , Células Cultivadas , Miócitos Cardíacos , Metabolismo , Patologia , RNA Mensageiro , Metabolismo , Taurina , Genética , Metabolismo
14.
Chinese Journal of Applied Physiology ; (6): 43-46, 2002.
Artigo em Chinês | WPRIM | ID: wpr-319385

RESUMO

<p><b>AIM</b>The characteristics of ryanodine receptor in rat cardiac sarcoplasmic reticulum (SR) and nuclear envelope (NE) were studied.</p><p><b>METHODS</b>Velocity and isopyknic gradient centrifugation was employed to fractionate rat SR and NE. Ryanodine receptor was assayed with [3H] ryanodine saturate binding to the preparations.</p><p><b>RESULTS</b>The maximal binding (Bmax) and dissociating constant (Kd) of ryanodine receptor in rat cardiac NE were, 1.7% and 60% of those in SR respectively. Phosphorylation in vitro by PKA and PKC increased Bmax of the receptors in SR by 372% and 121%, and augmented those in NE by 221% and 306%, without any effects on Kd.</p><p><b>CONCLUSION</b>Ryanodine receptors were present in rat myocardial NE, with lower density and higher affinity than those located in SR, which can be activated by PKA and PKC.</p>


Assuntos
Animais , Ratos , Cálcio , Metabolismo , Cinética , Miocárdio , Metabolismo , Membrana Nuclear , Metabolismo , Fisiologia , Fosforilação , Ratos Sprague-Dawley , Rianodina , Metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina , Metabolismo , Retículo Sarcoplasmático , Metabolismo , Fisiologia
15.
Chinese Journal of Applied Physiology ; (6): 51-54, 2002.
Artigo em Chinês | WPRIM | ID: wpr-319382

RESUMO

<p><b>AIM AND METHODS</b>To observe the effect of myocardial mitochondrial L-arginine (L-Arg)/nitric oxide (NO) system on mitochondrial Ca2+ transport by using purified rat mitochondria and incubation of them in vitro.</p><p><b>RESULTS</b>Compared with control group, incubation of mitochondria with L-Arg (10(-4) mol/L, NO substrate) or sodium nitroprusside (5 x 10(-7) mol/L, the donor of exogenous NO, SNP) increased significantly mitochondrial NO2- (66% and 89%, P < 0.01), respectively, and decreased the Ca2+ content (40% and 54%, P < 0.01). After L-Arg or SNP treatment, mitochondrial Ca2+ uptake were decreased by 67% and 85%, respectively (P < 0.01), vs control. The rate of mitochondrial Ca2+ release decreased by 11% and 8%, respectively (P < 0.01). When L-NAME (NO synthase inhibitor) was incubated with mitochondria and the L-Arg together, it inhibited the effects of L-Arg, NO2 on the mitochondrial NO2 formation, Ca2+ content descending, and decrease of Ca2+ uptake and release.</p><p><b>CONCLUSION</b>The data suggest that myocardial mitochondrial L-Arg /NO systems take part in the regulation of cardiomyocytes Ca2+ transportation.</p>


Assuntos
Animais , Feminino , Masculino , Ratos , Arginina , Metabolismo , Transporte Biológico , Cálcio , Metabolismo , Mitocôndrias Cardíacas , Metabolismo , Miócitos Cardíacos , Metabolismo , Óxido Nítrico , Metabolismo , Óxido Nítrico Sintase , Metabolismo , Ratos Wistar
16.
Acta Physiologica Sinica ; (6): 337-341, 2002.
Artigo em Chinês | WPRIM | ID: wpr-318989

RESUMO

To explore the changes in adrenomedullin (ADM) and receptor activity-modifying protein 2 (RAMP2) mRNA in myocardium and vessels in hypertension, a hypertensive rat model was prepared by administering L-NNA. Contents of ADM in plasma, myocardium and vessels were measured by radioimmunoassay (RIA). The levels of pro-ADM mRNA of myocardium and vessels were determined by competitive quantitative RT-PCR. The results showed that L-NNA induced hypertension and cardiomegaly. The ratio of heart to body weight increased by 35.5% (P<0.01). In hypertensive rats the ir-ADM in plasma, myocardium and vessels was increased by 80%, 72% and 57% (P<0.01), respectively compared with the control. The amounts of ADM mRNA in myocardium and vessels were increased by 50% and 109.2% (P<0.05), respectively, and the amounts of RAMP2 mRNA was increased by 132% and 87% (P<0.01), respectively, compared with control. The levels of ADM in myocardium and vessels were positively correlated with RAMP2 mRNA, the correlation coefficients were 0.741 and 0.885 (P<0.01), respectively. The results obtained indicate that in hypertensive rats, ADM is elevated in plasma, myocardium and ves-myocardium and vessel, and ADM and RAMP2 mRNA are up-regulated in myocardium and vessel. The ADM/RAMP2 system may play an important role in the pathogenesis of hypertension.


Assuntos
Animais , Ratos , Adrenomedulina , Metabolismo , Cardiomegalia , Metabolismo , Hipertensão , Metabolismo , Miocárdio , Metabolismo , Nitroarginina , Farmacologia , RNA Mensageiro , Proteína 2 Modificadora da Atividade de Receptores , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
17.
Acta Physiologica Sinica ; (6): 359-364, 2002.
Artigo em Inglês | WPRIM | ID: wpr-318985

RESUMO

The alterations of taurine transport and the expression of taurine transporter (TAUT) mRNA in myocardium and aortic wall were investigated in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. It was demonstrated that plasma taurine concentration and taurine release from myocardium and aortic wall in SHR were higher than those in WKY rats, whereas taurine content, taurine uptake and TAUT mRNA in myocardium and aortic wall of SHR were lower than those of WKY rats. In SHR, the maximal velocity (V(max)) of taurine transportation in myocardium and aortic wall was lower by 24% (P<0.05) and 35% (P<0.05) than that in WKY, their michaelis constants (Km) values were higher by 16% (P<0.05) and 39% (P<0.05), respectively. The results suggest that there is dysfunction of taurine transport in myocardium and aortic wall in SHR, which may be partly resulted from the decrease of TAUT activity and affinity, and down-regulation of TAUT gene expression.


Assuntos
Animais , Masculino , Ratos , Vasos Sanguíneos , Metabolismo , Proteínas de Transporte , Metabolismo , Coração , Técnicas In Vitro , Miocárdio , Metabolismo , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Taurina , Metabolismo
18.
Chinese Journal of Cancer Biotherapy ; (6)1995.
Artigo em Chinês | WPRIM | ID: wpr-582231

RESUMO

Objective: To investigate the effects of Interlukin- 4(IL-4) on the invasiveness and the expression of several cell surface antigens related to invasive and metastatic potentials of human hepatocellular carcinoma QGY-7701 cell line in vitro. Methods: QGY-7701 cells were incubated with high concentration of IL-4 or low concentration of IL-4 in different time. The expression of ICAM-1, CD44 and HLA-I was determined by fluorescence-activated cell sorter (FACS) analysis, the tumor cell binding affinity to extracellular matrix (ECM) components was measured by cell attachment assay, the degree of homotypic aggregation was quantified by cell aggregation assay. Results: IL-4 pretreatment can enhance the expression of ICAM-1 and HLA-I, suppress the expression of CD44 on hepatocellular carcinoma cell line and decrease the binding affinity to ECM components and the degree of homotypic aggregation of hepatocellular carcinoma cells. Conclusoin: IL-4 can inhibit the invasive and metastatic potentials of hepatocellular carcinoma cells.

19.
Chinese Journal of Endocrinology and Metabolism ; (12)1985.
Artigo em Chinês | WPRIM | ID: wpr-676346

RESUMO

The change in plasma ghrelin level after 4-and 12-week adjunctive therapy of rosiglitazones in type 2 diabetic patients inadequately controlled by sulphonylurea alone was observed and the relation between ghrelin and insulin resistance was analysed.The results showed that rosiglitazones significantly increased circulating ghrelin level and obviously decreased insulin resistance index after therapy for 4 and 12 weeks in type 2 diabetic patients.

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