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BACKGROUND AIMS: Refractory B-cell lymphomas are difficult to successfully treat with current chemotherapeutic regimens; however, immunotherapy may be an effective form of treatment for these patients. METHODS: Fourteen refractory lymphoma patients (age, 29-74 y) were enrolled in the trial. α-1,3-galactosyl (α-Gal) epitopes were synthesized on lymphoma cell membranes with the use of bovine recombinant α-1,3-galactosyltransferase (α-GT) and neuraminidase to enhance tumor immunogenicity. Subsequent incubation of processed cell membranes with autologous dendritic cells (DCs) in the presence of human serum containing abundant natural anti-α-Gal immunoglobulin G led to the effective phagocytosis of tumor membranes by DCs. The pulsed DCs and autologous cytokine-induced killer cells were then co-cultured to promote maximum cytotoxicity to lymphoma cells and were infused back into the donor lymphoma patients. Therapeutic responses were assessed by clinical observation, laboratory tests and a computed tomography scan at 6 months after treatment. RESULTS: Complete and partial remission occurred in four and three patients, respectively. The disease status remained unchanged in five patients, and disease progression was observed in two patients. No serious side effects or autoimmune diseases were observed in any participants. Serum lactate dehydrogenase and ß2-macroglobulin decreased in 11 and 14 patients, respectively. All patients showed robust systemic cytotoxicity in response to tumor lysate as measured by interferon-γ expression in peripheral blood mononuclear cells after treatment (P < 0.001). The number of peripheral immune effector cells (CD3(+)/CD4(+), CD8(+)/CD28(+) and CD16(+)/CD56(+) cells) increased significantly (P < 0.05) 3 months after treatment. CONCLUSIONS: Lymphoma cell-specific α-Gal immunotherapy is safe, effective and has great potential for the treatment of refractory B-cell lymphoma.
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Membrana Celular/metabolismo , Células Asesinas Inducidas por Citocinas/inmunología , Células Dendríticas/inmunología , Epítopos/inmunología , Galactosa/inmunología , Inmunoterapia/métodos , Linfoma de Células B/terapia , Adulto , Anciano , Animales , Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Bovinos , Citotoxicidad Inmunológica , Femenino , Citometría de Flujo , Humanos , Inmunoterapia/efectos adversos , Interferón gamma/metabolismo , Subgrupos Linfocitarios/inmunología , Linfocitos/inmunología , Linfoma de Células B/inmunología , Linfoma de Células B/patología , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
RATIONALE: Solitary fibrous tumor(SFT) is a rare and distinct clinical entity. There are few descriptions in the literature regarding the PET manifestations of SFTs. Herein,we report a case of multiple malignant SFT with PET/CT imaging findings and PET/Contrast Enhanced CT image findings. PATIENT CONCERNS: A 30-year-old woman presented with a history of a mass in neck increased gradually over 6 months without jaundice or other symptoms of obstruction.Serum laboratory results and tumor markers (AFP, CEA,CA199,CA724,CA153 and CA125)were normal. The whole body PET/CT scan showed lightly or mildly hypermetabolic and inhomogeneous metabolic which was different from other reports, and it was the first report of 18F-FDG-PET/CT findings of multiple malignant SFTs, which were confirmed by positive immunohistochemical staining for CD2-40, CD99, SMA and negative immunohistochemical staining for S100 and CD34. DIAGNOSES: She was diagnosed with multiple malignant solitary fibrous tumors which was confirmed by pathological results and 18F-FDG-PET/CT findings. INTERVENTIONS: The patient didn't receive any treatment because she was not suitable for surgery and refused any other therapy. OUTCOMES: The patient has been followed up for one year,and she was still alive. LESSONS: SFTs should be detected early and treated early.It was of high value in the diagnosis and differential diagnosis of SFTs by PET/CT imaging findings, which can not only identify the benign and malignant lesions, but also identify the lesion involved range.
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Tumores Fibrosos Solitarios/diagnóstico , Adulto , Biomarcadores de Tumor , Femenino , Fluorodesoxiglucosa F18 , Humanos , Tomografía Computarizada por Tomografía de Emisión de Positrones/métodos , Radiofármacos , Tumores Fibrosos Solitarios/diagnóstico por imagenRESUMEN
BACKGROUND: Despite new agent development and short-term benefits in patients with Colorectal Cancer (CRC), metastatic CRC cure rates have not improved due to high rates of oxaliplatin resistance and toxicity. There is an urgent need for effective tools to prevent and treat CRC and reduce morbidity and mortality of CRC patients. Exploring the effects of bioactive peptides on the antitumor to CRC was of vital importance to the clinical application. OBJECTIVE: This study aimed to investigate the therapeutic impact of Anticancer Bioactive Peptides (ACBP) on anticancer effect of oxaliplatin (LOHP) in human colorectal cancer xenografts models in nude mice. METHODS: HCT-116 cells were cultured in vitro via CCK-8 assays and the absorbance was measured at 450 nm. Apoptosis and cell cycle were assessed by Flow Cytometry (FCM) in vitro. HCT-116 human colorectal cancer cells inoculated subcutaneously in nude mice of treatment with PBS (GG), ACBP, LOHP, ACBP+LOHP (A+L) in vivo. The quality of life was assessed by dietary amount of nude mice, the weight of nude mice, inhibition rates, tumor weight and tumor volume. Immunohistochemistry and RT-qPCR method was conducted to determine the levels of apoptosisregulating proteins/genes in transplanted tumors. RESULTS: ACBP induced substantial reductions in viable cell numbers and apoptosis of HCT116 cells in combined with LOHP in vitro. Compared with the control GG group, ACBP combined low dose oxaliplatin (U) group demonstrated significantly different tumor volume, the rate of apoptosis, the expression levels of Cyt-C, caspase-3,8,9 proteins and corresponding RNAs (P<0.05). The expression of pro-apoptotic proteins in the cytoplasm around the nucleus was significantly enhanced by ACBP. Short term intermittent use of ACBP alone indicted a certain inhibitory effect on tumor growth, and improve the quality of life of tumor bearing nude mice. CONCLUSION: ACBP significantly increased the anti-cancer responses of low dose oxaliplatin (L-LOHP), thus, significantly improving the quality of life of tumor-bearing nude mice.
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Antineoplásicos/farmacología , Neoplasias Colorrectales/tratamiento farmacológico , Oxaliplatino/farmacología , Péptidos/farmacología , Animales , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias Colorrectales/patología , Femenino , Células HCT116 , Xenoinjertos , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Oxaliplatino/uso terapéutico , Péptidos/uso terapéuticoRESUMEN
OBJECTIVE: To explore the impact of anti-cancer bioactive peptide-S (ACBP-S) on cell proliferation, cell cycle and apoptosis in human stomach cancer cell line MGC-803 cells. METHODS: (1) The cultured MGC-803 cells were treated with ACBP-S at various concentrations for 24, 48, 72 h, respectively. The inhibition rate of proliferation of MGC-803 cells were evaluated by MTT assay. Cell apoptosis was observed by transmission electron microscopy. Cell cycle and apoptosis were analyzed by flow cytometry (FCM). RT-PCR was used to assay the changes of p27 mRNA expression. Immunocytochemistry was used to detect the changes of expression of p27, PCNA, Bax, Bcl-2 proteins, respectively. (2) a nude mouse xenograft model with gastric carcinoma cell was established. ACPB-S was administered into the tail vein of the mice in a dose of 7 microg, every other day, and the mice were killed after two weeks. The tumors were taken off for further analysis. RESULTS: (1) ACBP-S at concentrations of 5.0, 10.0 and 15.0 microg/ml inhibit the growth of MGC-803 cells in a concentration- and time-dependent manner. The concentration of ACBP-S at 20.0 microg/ml showed an inhibition rate of (86.6 + 0.1)%. Typical apoptotic changes were observed under the transmission electron microscope. The result of FCM in the range of 5.0 and 20.0 microg/ml for 24 h showed higher early apoptosis rates, (5.7 +/- 0.2)% and (13.9 +/- 0.6)%, respectively, with s significant difference compared with that of the control group (P < 0.05). The ratio of G0/G1 was significantly increased with the increase of incubation time at 20 microg/ml. RT-PCR showed that the expression of p27 mRNA in MGC-803 cells was markedly increased after ACBP-S treatment. (2) After ACBP-S administration the tumor growth in nude mice was inhibited by 34.2%. More apoptotic and necrotic cells were observed in the mice of treatment group by histological examination with HE staining. The immunocytochemistry demonstrated that the expression of Bax protein was significantly increased and Bcl-2 and PCNA protein expressions were significantly decreased after ACBP-S treatment. CONCLUSION: ACBP-S has marked inhibiting effect upon the growth of MGC-803 cells inducing more apoptosis. The anti-cancer mechanism is probably related with its regulatory effects on cell cycle and apoptosis in the tumor cells.
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Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Péptidos/farmacología , Neoplasias Gástricas/patología , Animales , Línea Celular Tumoral , Proliferación Celular , Inhibidor p27 de las Quinasas Dependientes de la Ciclina , Relación Dosis-Respuesta a Droga , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Péptidos/administración & dosificación , Antígeno Nuclear de Célula en Proliferación/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/metabolismo , Neoplasias Gástricas/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Anticancer bioactive peptide (ACBP) is extracted from normal goat spleens and exhibits antitumor activity alone and in combination with low cisplatin doses to achieve antitumor efficacy similar to higher cisplatin doses via sustained medication modes. In the present study, we investigated whether elevated levels of induced or normal ACBP in MKN45 gastric cancer (GC) cells may reduce their toxicity to oxaliplatin (LOHP) in a dosedependent manner. The growth inhibition rate (IR), morphological changes and gene expression were examined in MKN45 GC cells. Compared with normal ACBP, induced ACBP alone significantly enhanced the anticancer activity of LOHPmediated apoptosis and reduced the amount and sideeffects of LOHP (P<0.05). The inhibition of cancer cell growth at high concentrations of induced ACBP and LOHP was significantly more effective than at low concentrations. In addition, for the first time, we examined the potential of a combination of induced ACBP and LOHP to increase LOHP sensitivity in human gastric carcinoma xenograft tumors. Nude mice were implanted with human gastric carcinoma MKN45 cells and treated with an intraperitoneal injection of 0.5 ml of normal saline, 30 µg/ml ACBP, 20 µg/ml LOHP or 30 µg/ml ACBP + 20 µg/ml LOHP [combination of anticancer bioactive peptide and oxaliplatin (A+L)] via the tail vein twice a week. In vivo shortterm intermittent use of induced ACBP alone significantly inhibited MKN45 tumor growth. The combination of induced ACBP and LOHP also significantly improved the quality of life of the nude mice and reduced the toxicity of LOHP. Based on flow cytometry and gene expression analyses, A+L significantly increased the proportion of cells in the G2/M phase (P<0.05) relative to ACBP or LOHP alone, and shortterm intraperitoneal injection of ACBP increased the sensitizing effect of LOHP. Collectively, these results suggest that high levels of induced ACBP in combination with LOHP via a shortterm intermittent medication mode could be a useful clinical therapeutic strategy for GC.
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Apoptosis/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Compuestos Organoplatinos/farmacología , Fragmentos de Péptidos/farmacología , Neoplasias Gástricas/tratamiento farmacológico , Animales , Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Oxaliplatino , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Objectives. In order to enhance the immunity of cancer patients to prevent relapse or to prolong survival time, umbilical cord blood mononuclear cells (UCMCs) were transplanted to cancer patients. Patients and Methods. UCMCs were transfused to 63 immunocompromised gastrointestinal cancer patients with nonmyeloablative (NMA) conditioning regimen. Results. The clinical study showed that the number of both T and B cells increased much more rapidly after transfusion of UCMCs than that of the control group without transplantation (p < 0.01). Proinflammation cytokines IFNγ and TNFα in serum increased to or above the normal range in 80.9% of patients at 12 weeks after UCMC transfusion. However, they recovered to the normal range in 21.7% of patients at the same time point in the control group only. In addition, the clinical investigation also showed that the transfusion of UCMC increased stable disease (SD) and reduced progressive disease (PD) significantly (p < 0.01); however, it did not have significant effects on complete response (CR), partial response (PR), or mortality rates compared with the control group (p > 0.05). Conclusions. UCMCs have powerful repairing effects on damaged cells and tissues and may reconstruct the impaired immunity. Transfusion of UCMCs could reconstruct the immunity of cancer patients with immunosuppression.
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Sangre Fetal/trasplante , Neoplasias Gastrointestinales/terapia , Leucocitos Mononucleares/trasplante , Recurrencia Local de Neoplasia/terapia , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Citometría de Flujo , Neoplasias Gastrointestinales/sangre , Neoplasias Gastrointestinales/inmunología , Neoplasias Gastrointestinales/patología , Humanos , Interferón gamma/inmunología , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/sangre , Recurrencia Local de Neoplasia/inmunología , Recurrencia Local de Neoplasia/patología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
AIM: To determine whether Helicobacter pylori (H pylori) vacuolating cytotoxin (VacA) regulates release of pro-inflammatory cytokines (IL-1beta, IL-8, TNF-alpha, and IL-6) or alters gastric epithelial cell viability and to determine whether NaCl affects these VacA-induced changes. METHODS: Vacuolating activity was determined by measuring the uptake of neutral red into vacuoles of VacA-treated human gastric epithelial (AGS) cells. AGS cell viability was assessed by direct cell counting. Specific enzyme-linked immunosorbent assays (ELISA) and reverse transcriptase-polymerase chain reaction(RT-PCR) were performed to examine the effects of H pylori VacA and NaCl on cell pro-inflammatory cytokine production in AGS cells. Immunohistochemical staining of gastric tissue from Mongolian gerbils was used to confirm VacA-induced pro-inflammatory cytokine production and the effects of NaCl on this VacA-induced response. RESULTS: Addition of VacA alone reduced AGS cell viability (P < 0.05), and this reduction was enhanced by high doses of NaCl (P < 0.05). VacA alone induced expression of TNF-alpha, IL-8 and IL-1beta, while NaCl alone induced expression of TNF-alpha and IL-1beta. Changes in mRNA levels in the presence of both VacA and NaCl were more complicated. For the case of TNF-alpha, expression was dose-dependent on NaCl. IL-6 mRNA was not detected. However, low levels of IL-6 were detected by ELISA. Positive immunohistochemical staining of IL-1, IL-6, and TNF-alpha was found in gastric tissue of H pylori-infected gerbils fed with either a normal diet or a high salt diet. However, the staining of these three cytokines was stronger in H pylori-infected animals fed with a 5 g/kg NaCl diet. CONCLUSION: VacA decreases the viability of AGS cells, and this effect can be enhanced by NaCl. NaCl also affects the production of pro-inflammatory cytokines induced by VacA, suggesting that NaCl plays an important role in H pylori-induced gastric epithelial cell cytotoxicity.
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Proteínas Bacterianas/toxicidad , Citocinas/genética , Mucosa Gástrica/efectos de los fármacos , Cloruro de Sodio/farmacología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Citocinas/biosíntesis , Mucosa Gástrica/citología , Mucosa Gástrica/inmunología , Gerbillinae , Humanos , Masculino , ARN Mensajero/análisis , Vacuolas/efectos de los fármacosRESUMEN
BACKGROUND: The World Health Organization reported in 2011 that irrational use of medicines was a serious global problem that is wasteful and harmful. The worst is use of ineffective or harmful interventions which should not be used at all. However, little is known about the changes that 20 years of evidence-based medicine has made particularly in reducing use of ineffective interventions. We surveyed clinicians in China to show how often ineffective interventions were still used in practice. METHODS: 3,246 clinicians from 24 tertiary hospitals were surveyed in person and another 3,063 through an online survey between 2006-2007. The main outcomes are prescription by a clinician, and use in a patient of, an ineffective intervention and of a matched effective intervention in patients with the same disease. 129 ineffective interventions for 68 diseases were identified from the BMJ Clinical Evidence and included in the survey. One effective intervention was identified for each disease and a total of 68 effective interventions were thus also included. The frequency of use of effective interventions was used as a reference for that of ineffective intervention. RESULTS: The mean prescription rate by clinicians is 59.0% (95% confidence interval (95% CI): 58.6% to 59.4%) and 81.0% (95% CI: 80.6% to 81.4%) respectively for ineffective and effective interventions. The mean frequency of use in patients is 31.2% (95% CI: 30.8% to 31.6%) and 56.4% (95% CI: 56.0% to 56.8%) for ineffective and effective interventions respectively. The relative reduction in use of ineffective interventions as compared with that of matched effective interventions is 27.2% (95% CI: 27.0% to 27.4%) and 44.7% (95% CI: 44.3% to 45.1%) for clinician's prescription and use in patients respectively. 8.6% ineffective interventions were still routinely used in practice. CONCLUSIONS: Ineffective interventions were still commonly used. Efforts are necessary to further reduce and eventually eliminate ineffective interventions from practice.
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Evaluación de Resultado en la Atención de Salud , China , Estudios Transversales , Femenino , Humanos , Masculino , Resultado del TratamientoRESUMEN
The role of hypoxia-inducible factor-1 (HIF-1) in tumor development and progression is well-established but its effect on tumor growth remains controversial. The present study investigated the effect of HIF-1 on tumor growth using the estrogen receptor-negative breast cancer cell line, MDA-MB-231. Using Western blotting, we detected a higher level of HIF-1α protein in MDA-MB-231 cells than in any other breast cancer cell lines analyzed, and this was accompanied by a more rapid growth pattern. Interruption of HIF-1α expression using small interference RNA (siRNA) significantly suppressed cell growth and increased apoptosis, but the cell cycle was not affected. Activated fragments with increased caspase 3 activity and a mobility shift of B cell lymphoma (Bcl-2) were also detected in cells treated with HIF-1α siRNA. HIF-1 allows breast cancer cells to grow under long-term serum deprivation by inactivation of the caspase cascade and thus inhibition of apoptosis. Blocking HIF-1α protein resulted in loss of Bcl-2 function, which may contribute to the activation of the caspase cascade.
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VacA is a vacuolation protein toxin secreted by Helicobacter pylori. Many compounds have been implicated in the regulation of VacA toxin activity. In this study, regulation of cell vacuolation induced by VacA was observed with the addition of glycine, glycine hydrochloride, xylitol, and taurine by neutral red dye uptake assay using gastric human epithelial cell cultures. Glycine, xylitol, and taurine increased cell vacuolation significantly after 48h (p<0.05), with their effect apparent in a wide concentration range (0.2mM to about 100mM). Changes were sharp in respect of concentration and showed little dose-response characteristics. In contrast, upregulation of glycine hydrochloride on cell vacuolation in weak acidic extracellular pH was much retarded with VacA activity not initiated until 72h. In addition, our results showed that cell vacuolation was highest when the pH was 6.8. The increase in vacuolation was gentle in weak acidic extracellular pH and the increase dose-dependent with a Pearson correlation coefficient (r) of 0.986 from 0.2 to 6.25mM. In this concentration range and at the same time point, the pH decrease was negatively correlated with vacuolating activity (r=0.922, p<0.01). In conclusion, our study showed that three small molecular compounds can increase vacuolation induced by VacA toxin in vitro.
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Proteínas Bacterianas/toxicidad , Toxinas Bacterianas/toxicidad , Glicina/farmacología , Taurina/farmacología , Vacuolas/efectos de los fármacos , Xilitol/farmacología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Rojo Neutro/metabolismo , Vacuolas/metabolismoRESUMEN
OBJECTIVE: To develop a sensitive and specific RT-PCR assay using the mRNA of MAGE-1 and MAGE-3 genes as specific tumor markers for the detection of the tumor cells in the peripheral blood of patients with gastric cancer. METHODS: Peripheral blood was obtained from 40 patients with gastric cancer and from 20 healthy volunteers. The mRNA of MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMC) was detected by RT-PCR. The expressions of MAGE-1 and MAGE-3 mRNA in the tumor tissues of these gastric cancer patients were also detected by RT-PCR. Meanwhile,CEA expression by nested RT-PCR in PBMC of 40 gastric cancer patients was also detected. RESULTS: Of 40 gastric cancer patients, MAGE-1 and MAGE-3 mRNA were positive in 47.5% (19/40) and 25% (10/40) of PBMC respectively, and in 62.5% (25/40) and 30% (12/40) of gastric cancer tissues respectively. As a whole, in the PBMC of 40 gastric cancer patients, 25 (62.5%) samples were found to express at least one type of MAGE mRNA. In the patients whose tumors did not express MAGE-1 and/or MAGE-3 genes, the corresponding MAGE mRNA was also undetected in their PBMC. There was no expression of MAGE-1 or MAGE-3 gene in the PBMC from the 20 healthy donors. The positive rate of MAGE mRNA in PBMC was closely correlated with the tumor stage and lymph node metastasis (P <0.05). Positive rate of CEA gene expression was 32.5% (13/40) in the PBMC of 40 gastric cancer patients, 29 (72.5%)samples were detected to express at least one type of MAGE gene and CEA gene mRNA. CONCLUSIONS: MAGE-1, MAGE-3 and CEA mRNA are specifically detected with high percentage in the PBMC of gastric cancer patients by RT-PCR. They could be used as specific tumor markers for the detection of the circulating gastric cancer cells, and the detection results may be helpful to evaluate the prognosis of gastric cancer patients.
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Antígenos de Neoplasias/sangre , Biomarcadores de Tumor/sangre , Proteínas de Neoplasias/sangre , Neoplasias Gástricas/sangre , Adulto , Anciano , Antígenos de Neoplasias/genética , Antígeno Carcinoembrionario/sangre , Femenino , Humanos , Masculino , Antígenos Específicos del Melanoma , Persona de Mediana Edad , Proteínas de Neoplasias/genética , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologíaRESUMEN
A novel method for the determination of monoethylglycinexylidide (MEGX) (lidocaine metabolin) in serum using solid phase extraction (SPE) and capillary gas chromatography-mass spectrometry (GC-MS) was established. The serum sample was extracted with a CN-SPE column. An HP-5MS capillary column (15 m x 0.25 mm x 0.1 microm) was used. The initial temperature of the column was set at 100 degrees C, held for 1 min, then raised to 200 degrees C at 40 degrees C/min, and held at 200 degrees C for 0.5 min. The sample size was 2 microL, and the split ratio was set at 1 : 1. The carrier gas was high purity helium with a flow rate of 1.0 mL/min. The monitoring ions for the determination were m/z 58 for MEGX and m/z 86 for procaine (internal standard). The calibration curve of MEGX had good linear relationship in the range of 1.562 - 25 ng/mL ( r = 0.998 1). The limit of detection was 0.5 ng/mL. The extraction recovery ranged from 80.1% to 85.7%. The method advanced the quantitative analysis of MEGX in serum by combining rapid and efficient SPE with specific and sensitive quantitation by GC-MS.