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Objective: To investigate the effect of hypoxia inducible factor 2α (HIF-2α) on regulating CUB domain-containing protein 1 (CDCP1) and its role in hepatocellular carcinoma metastasis. Methods: HIF-2α-knocked down and HIF-2α-stably overexpressing cells (MHCC97H) were prepared by small interfering RNA (siRNA) and lentivirus transfection, respectively. The expression of CDCP1 protein and mRNA in the above cells was detected by western blot and real-time PCR. The effect of HIF-2α on cell invasion ability was determined by Transwell assay. Furthermore, immunohistochemical staining was performed to detect the expression of CDCP1 in human HCC tissue samples. Results: Both HIF-2α and CDCP1 were induced under hypoxic conditions. The activation of CDCP1 under hypoxic conditions was dependent on the expression of HIF-2α.When HIF-2α was overexpressed, the mRNA level of CDCP1 was greatly upregulated (5.92±0.28, P<0.05). When HIF-2α was knocked down by siRNA for 48 h and 72 h, the expression of CDCP1 was significantly downregulated (48 h: 0.25±0.04; 72 h: 0.18±0.02, all P<0.05). Moreover, analysis of human HCC samples showed that CDCP1 expression was correlated with tumor-free survival (P<0.05). Conclusions: The results of this study indicate that the expression of CDCP1 is regulated by HIF-2α and is correlated with the progression of HCC. Inhibition of HIF-2α/CDCP1 may play certain inhibitory role in the metastasis of HCC.
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Antígenos CD/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Carcinoma Hepatocelular/secundario , Moléculas de Adhesión Celular/metabolismo , Neoplasias Hepáticas/patología , Proteínas de Neoplasias/metabolismo , Animales , Antígenos CD/genética , Antígenos de Neoplasias , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Carcinoma Hepatocelular/metabolismo , Moléculas de Adhesión Celular/genética , Movimiento Celular , Regulación hacia Abajo , Técnicas de Inactivación de Genes , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Neoplasias Hepáticas/metabolismo , Ratones Desnudos , Invasividad Neoplásica , Proteínas de Neoplasias/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño , Reacción en Cadena en Tiempo Real de la Polimerasa , Transfección/métodos , Hipoxia Tumoral , Regulación hacia ArribaRESUMEN
Objective: To find a suitable method for the determination of oxalic acid in the urine of patients with stones, in order to provide a new method and basis for the prevention and treatment of stone. Methods: Liquid chromatography combined with tandem mass spectrometry was used to analyze oxalic acid in urine directly.The accuracy, stability, repeatability and other indicators of the results were tested. Results: The results showed a good linear relationship with the concentration of oxalic acid in urine. y=58.524x-15.246 (R(2)=0.979 02). The results were stable, reproducible (the intra-day and inter-day coefficient of variation was less than 10% and 15%, respectively), and the accuracy was comparable with that of the enzyme method (N=20, R=0.93). Conclusion: Using the method of this study to detect the content of oxalic acid in urine has the advantages of simple operation, good repeatability, accurate results, and low price. It is worth to be popularized and applied in clinical practice.
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Espectrometría de Masas en Tándem , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Humanos , Ácido Oxálico , Reproducibilidad de los ResultadosRESUMEN
Objective: To find a suitable method for the determination of citric acid in the urine of patients with stones, in order to provide a new method and basis for the prevention and treatment of stone. Methods: Liquid chromatography tandem mass spectrometry was used to analyze the citric acid in urine directly. And the accuracy, stability, repeatability and other indicators of the results were detected. Results: The results showed a good linear relationship with the concentration of citric acid in urine. y=50.31x+ 0.002 6 (R(2)=0.994 21). The results were stable, reproducible [intra-day (Coefficient of Variance) CV ≈1% and inter-day CV<10%], and the accuracy of which was comparable with that of the enzyme method (n=20, R=0.97). Conclusion: Using the method of this study to detect the content of citric acid in urine has the advantages of simple operation, good repeatability, accurate results, and low price. So it is worth to be popularized and applied in clinical practice.
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Ácido Cítrico/orina , Cálculos Renales , Espectrometría de Masas en Tándem , Cromatografía Liquida , Humanos , Reproducibilidad de los ResultadosRESUMEN
This study evaluated the feasibility and effectiveness of using the bispectral index (BIS) to monitor anesthetic depth in patients with severe burns receiving intravenous target-controlled infusion (TCI) of remifentanil and propofol. We randomly assigned 80 patients undergoing elective escharectomy (<1 week) to BIS (A) and control (B) groups. All patients received remifentanil and propofol as intravenous TCI anesthesia. Clinical data were recorded at different time points. The time from drug withdrawal to eye opening upon the patient hearing his/her name called and upon reaching an Aldrete score of 9 points was also recorded. During anesthesia maintenance, the target concentrations of remifentanil and propofol in group A were significantly lower than that in group B (2.12 ± 0.35 vs 2.50 ± 0.21 ng/mL and 2.54 ± 0.22 vs 2.86 ± 0.31 µg/mL, respectively; P < 0.01). The time from drug withdrawal to eye opening upon the patient hearing his/her name called and reaching an Aldrete score of 9 points in group A was considerably shorter than that in group B (7.90 ± 0.58 vs 8.35 ± 0.66 min and 9.15 ± 0.69 vs 11.13 ± 0.96 min, respectively; P < 0.01). In both groups, mean arterial pressure and heart rate values at each time point after loss of consciousness were significantly lower than the baseline values (P < 0.05), with the exception of 2 min after intubation. The use of BIS to monitor anesthetic depth in patients with severe burns receiving TCI of remifentanil and propofol during the perioperative period reduces propofol consumption and shortens the consciousness recovery time in patients.
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Anestesia , Quemaduras/cirugía , Monitores de Conciencia , Piperidinas/farmacología , Propofol/farmacología , Adulto , Periodo de Recuperación de la Anestesia , Presión Sanguínea/efectos de los fármacos , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Humanos , Infusiones Intravenosas , Masculino , Atención Perioperativa , Piperidinas/administración & dosificación , Propofol/administración & dosificación , Remifentanilo , Vigilia/efectos de los fármacosRESUMEN
We investigated the therapeutic effect of Xin Mai Jia (XMJ) on atherosclerosis (AS) in rats. Rat models of AS were established by peritoneally injecting vitamin D, feeding a high-fat diet, and inducing balloon injuries in rats. The stomachs of the rats were irrigated continuously for 10 weeks with XMJ. Blood lipid- and hemorheology-related indices of blood samples were detected. Pathological changes in the right common carotid arterial tissues were also determined. The protein expression levels of endothelial nitric oxide synthase, angio-tensin-1, and endothelin-1 were determined by western blotting. XMJ reduced cholesterol, trigylecride, and low-density lipoprotein levels as well as blood viscosity, sedimentation, and hematocrit. Furthermore, XMJ alleviated vascular endothelial injury and reduced/eliminated atherosclerotic plaques. In contrast, XMJ significantly increased the endothelium-dependent relaxing response of the AS rat models. The western blotting results showed that XMJ upregulated endothelial nitric oxide synthase but downregulated angiotensin-1 and endothelin-1. XMJ prevented the development of AS by regulating blood lipid levels, hemorheology, and vascular function.
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Aterosclerosis/sangre , Aterosclerosis/tratamiento farmacológico , Colesterol/sangre , Medicina Tradicional China , Angiotensinas/biosíntesis , Angiotensinas/sangre , Animales , Aterosclerosis/inducido químicamente , Dieta Alta en Grasa , Endotelina-1/biosíntesis , Endotelina-1/sangre , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Expresión Génica , Humanos , Lipoproteínas LDL/sangre , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/biosíntesis , Óxido Nítrico Sintasa de Tipo III/sangre , Ratas , Vitamina D/toxicidadRESUMEN
OBJECTIVE: p38 mitogen-activated protein kinase (p38MAPK) is a critical negative regulator for nucleus pulposus (NP) cell metabolism contributing to intervertebral disc degeneration (IDD). Histone deacetylase 4 (HDAC4) has the ability to mediate cell proliferation and is affected by p38MAPK. It is unclear whether the p38 MAPK inhibitor SD0006 (SD) can regulate IDD. Our study aims to explore the effect of SD in the progression of IDD, as well as its potential mechanism. PATIENTS AND METHODS: NP cells isolating from mild degenerated NP tissues were cultured, and IL-1ß or Asiatic acid (AA) was used to activate p38MAPK and accelerate the NP cell degradation. Then, SD was used to reject the p38MAPK activation. After that, the levels of phosphorylated p38MAPK (p-p38), HDAC4, proliferating cell nuclear antigen (PCNA), inflammatory factor, proliferative cell rate, and cell cycle were determined by Western blot, RT-PCR, flow cytometry, and immunofluorescence, respectively. RESULTS: The results showed that the activation of p38MAPK by IL-1ß and AA decreased the HDAC4 expression, affected the collagen-â ¡ expression, upregulated the TNF-α, IL-6, MMP-3, and ADAMTS mRNA levels, and prevent the NP cell proliferation by mediating cell cycles. However, the application of SD alleviated the negative effect of p-p38 by upregulating HDAC4, anti-inflammation, and promoting cell proliferation, while the blocking of HDAC4 expression partly abolished the effect of SD. CONCLUSIONS: SD can prevent NP cell degeneration by promoting cell proliferation and suppressing inflammation via p38MAPK/HDAC4 pathway.
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Histona Desacetilasas/metabolismo , Núcleo Pulposo/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Pirazoles/farmacología , Pirimidinas/farmacología , Proteínas Represoras/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Humanos , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
This study is an extension of our previous report that alloimmunization produces a serum factor that suppresses the lymphocytotoxic antibody response. The experiments presented herein show that: (1) serum and IgG of pretreated rats contain an antibody directed against the constant region of IgG; (2) the anti-IgG has strong anti-Fc-gamma and weak anti-Fab activity; (3) the antibody also reacts with IgM; (4) the antiimmunoglobulin is an autoantibody; and (5) the antiimmunoglobulin suppresses the primary B cell response in vitro.
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Anticuerpos Antiidiotipos/fisiología , Linfocitos B/inmunología , Tolerancia Inmunológica/inmunología , Inmunoglobulina G/inmunología , Isoanticuerpos/inmunología , Factores Supresores Inmunológicos/inmunología , Animales , Inmunización , Fragmentos Fab de Inmunoglobulinas/metabolismo , Fragmentos Fc de Inmunoglobulinas/metabolismo , Masculino , Ratas , Ratas Endogámicas BN , Ratas Endogámicas LewRESUMEN
To demonstrate whether anti-IgG autoantibodies of different isotypes and specificities play a role in kidney transplantation, pretransplant sera of recipients (a) with well-functioning grafts (n = 40); (b) with reversible rejection episodes that were treated successfully (n = 63); and (c) with irreversible graft rejection (n = 40) were tested for four different anti-IgG autoantibody activities. A protective effect of IgG-anti-F(ab)2 gamma antibodies on graft survival was observed (p less than 0.01). High pretransplant IgG-anti-Fc gamma activity was found to be associated with a low kidney graft survival rate (p less than 0.02). Pretransplant IgM-anti-F(ab)2 gamma and IgM-anti-Fc gamma activities showed no effect on kidney graft outcome.
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Anticuerpos Antiidiotipos/inmunología , Autoanticuerpos/inmunología , Inmunoglobulina G/inmunología , Trasplante de Riñón/inmunología , Rechazo de Injerto/inmunología , Humanos , Técnicas para Inmunoenzimas , Fragmentos Fab de Inmunoglobulinas/inmunología , Isotipos de Inmunoglobulinas/inmunología , Inmunoglobulina M/inmunología , PronósticoRESUMEN
Treatment of LEW rats before transplantation with BN blood cells preincubated with LEW-anti-BN serum resulted in prolonged BN kidney graft survival (untreated, 8 +/- 0.4 days; pretreated, 124 +/- 36). The serum of pretreated animals contains a factor which suppresses T cell proliferation against donor antigens. This effect was not mediated by antiidiotypic antibodies because it was reproduced in a donor-unrelated system. The serum and IgG of pretreated animals also suppressed the humoral immune response. As for T cells, the antibody-suppressive effect was not donor-restricted. A broadly reactive anti-immunoglobulin (anti-B cell) autoantibody was found in the IgG fraction of immunized animals. The mediation of immunosuppression by broadly reactive anti-B and T cell autoantibodies induced by the immunization with antibody-coated blood cells is discussed.
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Autoanticuerpos/inmunología , Células Sanguíneas/trasplante , Supervivencia de Injerto/inmunología , Inmunización , Terapia de Inmunosupresión , Trasplante de Riñón/inmunología , Animales , Anticuerpos Antiidiotipos/inmunología , Linfocitos B/inmunología , Células Sanguíneas/inmunología , Inmunoglobulina G/inmunología , Activación de Linfocitos/inmunología , Ratas , Ratas Endogámicas BN , Ratas Endogámicas Lew , Factores Supresores Inmunológicos/sangre , Linfocitos T/inmunologíaRESUMEN
We studied the pharmacokinetics and distribution of homoharringtonine (HHT), an antitumor alkaloid, in anesthetized dogs using chromatographic and radiochemical techniques. Uniformly tritiated HHT was administered i.v. to five dogs at doses of 0.05 to 0.34 mg/kg, 200 microCi per animal. Unchanged HHT disappeared in a triphasic manner from the plasma with an initial plasma t1/2 of 9.4 +/- 4.2 min, an intermediary t1/2 of 1.4 +/- 0.5 h, and a terminal t1/2 of 40.6 +/- 4.6 h. The plasma clearance was 114.0 +/- 20.1 ml/kg-1 h-1 and the steady-state volume of distribution was 6.2 +/- 0.7 1/kg. In 72 h, 40.1% +/- 4.0% of the administered radioactivity was excreted in the urine, 17.8% +/- 2.7% of which was unchanged HHT. HHT was metabolized extensively to one major and two minor metabolites. Biliary excretion of total radioactivity was 14.4% in 5 h, 2% of which was HHT. HHT concentration in the CSF was highest 4 h after drug administration, about 40% of the concentration in the concurrent plasma. At autopsy 5 h after dosing, the highest percentage of HHT was in the liver (7.4%), followed by the small intestine (2.5%), stomach (1.0%), pancreas (0.8%), kidneys (0.8%), and lungs (0.7%). The heart, spleen, large intestine, and brain each retained less than 0.5%. However, 24 h after dosing, 4% of the HHT still remained in the liver, 1% in the small intestine, and less than 1% in the other organs. HHT seems to be extensively metabolized in dogs and partially retained in the body.
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Alcaloides/farmacocinética , Antineoplásicos/farmacocinética , Harringtoninas/farmacocinética , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/líquido cefalorraquídeo , Bilis/metabolismo , Perros , Femenino , Mucosa Gástrica/metabolismo , Harringtoninas/administración & dosificación , Harringtoninas/líquido cefalorraquídeo , Homoharringtonina , Infusiones Intravenosas , Intestino Delgado/metabolismo , Riñón/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Masculino , Páncreas/metabolismo , Distribución TisularRESUMEN
Synthetic corticotropin-releasing factor (CRF) was administered pre-operatively to five patients who underwent craniotomy for brain tumors, acute subdural and intracerebral hematomas. After craniotomy, an intraventricular catheter is placed in the ventricle, on the normal side of brain. This is for intracranial pressure monitoring. The data obtained were pooled and compared with the saline group (n = 6). The CRF group was found to have statistically significant difference (P less than 0.01). This preliminary study showed that CRF decreases postoperative edema.
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Edema Encefálico/tratamiento farmacológico , Hormona Liberadora de Corticotropina/uso terapéutico , Complicaciones Posoperatorias/tratamiento farmacológico , Adulto , Anciano , Anciano de 80 o más Años , Astrocitoma/cirugía , Neoplasias Encefálicas/cirugía , Femenino , Hematoma Subdural/cirugía , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Angiotensin II (Ang II)-induced apoptosis was demonstrated for the first time in cultured adult rat ventricular myocytes (ARVMs) isolated by retrograde heart perfusion with Krebs-Henseleit bicarbonate (KHB) buffer containing collagenase and hyaluronidase. ARVMs incubated with 10 mumol/L Ang II for 48 h showed morphological features of apoptosis (cellular shrinkage, condensation of cytoplasm) and a characteristic "ladder" of DNA bands representing integer multiples of the internucleosomal DNA length about 180-200 bp, which became more evident with further incubation up to 72 h. With shorter incubation time (< or = 24 h) or at a lower Ang II concentration (< 10 mumol/L), such changes failed to occur. This effect of Ang II could be abolished by losartan (10 mumol/L), verapamil (1 mumol/L) or staurosporine (10 nmol/L). The above results indicate that Ang II-induced apoptosis in ARVMs may be mainly mediated by Ang II type I (AT1) receptors with [Ca2+]i and protein kinase C (PKC) playing a critical role.
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Angiotensina II/farmacología , Apoptosis/efectos de los fármacos , Miocardio/citología , Animales , Células Cultivadas , Femenino , Ventrículos Cardíacos , Masculino , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptores de Angiotensina/metabolismoRESUMEN
In the present study, the effect of high density lipoproteins (HDLs) on the antiaggregating activity of nitric oxide (NO) derived from endothelial cells was investigated with the use of cultured bovine aortal endothelial cells (BAECs). The BAECs were placed in an aggregometer in contact with rabbit platelets after blocking cyclo-oxygenase with acetylsalicylic acid. Under this circumstance, the antiaggregating effect of endothelial cells was exclusively dependent on the release of NO, which was further confirmed by prevention of antiaggregating activity of BAECs with 1 mmol/L N(G)-Nitro-L-arginine. When this system was used, thrombin (0.1 U/ml) evoked 67.33+/-7.52% aggregation of rabbit platelets (2 10(8)/ml). This effect was inhibited by NO derived from endothelial cells (1 10(5) 1 10(6)/ml) in a cell number dependent manner. HDLs (1 mg/ml), added into the system immediately before BAECs, enhanced this antiaggregating effect of NO. However, incubating BAECs with HDLs for an hour and removing the HDLs by centrifugation did not have the same effect, unless HDLs were present during aggregation. No direct effect of HDLs on platelet aggregation was observed. The above findings suggest that HDLs can enhance the antiaggregating effects of BAECs mediated by direct interaction with NO.
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Endotelio Vascular/metabolismo , Lipoproteínas HDL/farmacología , Óxido Nítrico/fisiología , Agregación Plaquetaria , Animales , Aorta/citología , Bovinos , Células Cultivadas , Endotelio Vascular/citología , ConejosRESUMEN
Apyrase activities in some tissues and cells, such as peripheral vascular endothelial cells, have been reported, but these in endocardium endothelial cells have not been reported. The present study was to characterise the properties of bovine endocardium endothelial cells (BEEC)-associated apyrase. Apyrase activity was assayed by inorganic phosphate release, which could be inhibited concentration-dependently by NaN3, an apyrase inhibitor. NaF (20 mmol/L), another inhibitor of apyrase, also markedly inhibited the activity. EDTA or EGTA (1 mmol/L) could also inhibit the activity completely. However, the inhibitor for Na+/K(+)-ATPase, ouabain (3 mmol/L) did not affect the enzyme activity. BEEC apyrase activity was dependent on divalent cations (Ca2+ or Mg2+) and pH value.
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Apirasa/metabolismo , Endocardio/enzimología , Animales , Apirasa/antagonistas & inhibidores , Calcio/metabolismo , Bovinos , Células Cultivadas , Endocardio/citología , Endotelio/citología , Endotelio/enzimología , Concentración de Iones de Hidrógeno , Magnesio/metabolismo , Azida Sódica/farmacologíaRESUMEN
OBJECTIVE: To clarify the mechanism of Yiqi Tongluo Pill (YQTLP) in treating coronary heart disease and hypertension. METHODS: The clinical effects of YQTLP on 97 coronary heart diseases (CHD) and hypertension patients, in comparing to the 92 patients treated with nitroglycerin was investigated. The changes of plasma lipid peroxide (LPO), superoxide dismutase (SOD), tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) and nitric oxide (NO) were observed in the patients treated with YQTLP and nitroglycerin, which were compared to 30 healthy subjects. RESULTS: The levels of LPO and PAI increased and the levels of SOD, t-PA and NO decreased significantly in the patients. The clinical effects of YQTLP were better than that of nitroglycerin in the patients with unstable angina, while the effects of YQTLP were worse than that of nitroglycerin in the patients of hypertension and hypertension with CHD. YQTLP could decrease the levels of LPO and PAI and increase the levels of SOD, t-PA and NO significantly. The effects of YQTLP on t-PA and SOD were better than that of nitroglycerin. CONCLUSION: The protective mechanisms of YQTLP were related to inhibit lipid peroxidation, protect endothelium-derived relaxing factor and adjust the fibrinolytic activities.
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Enfermedad Coronaria/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Fibrinólisis , Hipertensión/tratamiento farmacológico , Óxido Nítrico/sangre , Adulto , Anciano , Enfermedad Coronaria/sangre , Femenino , Fibrinólisis/efectos de los fármacos , Humanos , Hipertensión/sangre , Peróxidos Lipídicos/sangre , Masculino , Persona de Mediana Edad , Superóxido Dismutasa/sangre , Activador de Tejido Plasminógeno/sangreRESUMEN
OBJECTIVE: To establish a hairy root culture system by double transformation for Trichosanthes kirilowii. METHOD: 1. Crown galls were induced by direct infection of sterile seedlings of T. kirilonii with Agrobacterium tumefaciens C58, and then the hairy roots were obtained from the regenerated plants by infection with A. rhzogenes 15834; 2. Transformation of Ti and Ri plasmids was inspected by high-pressure-paper electrophoresis; 3. The protein contents in the tissues of T. kirilowii were inspected by spectrophotometer and SDS-PAGE. RESULT: A hairy root culture system has been established successfully by double transformation with Ti and Ri plasmids in T. kirilowii. CONCLUSION: Compared with the ordinary hairy roots, the double transformed hairy roots grow faster but retain similar protein contents.