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Objective: To establish a method for the rapid determination of acetaminophen (APAP) in human plasma by LC-MS/MS. Methods: The plasma samples were extracted by methanol and acetonitrile (1: 1) and purified directly. C(18) column was used for sample separation. The mobile phase were methanol (5 mmol/L ammonium acetate) and water (5 mmol/L ammonium acetate). Samples were analyzed by LC MS/MS with the electrospray ionization multi reaction monitoring (MRM) mode. Results: The calibration curves of APAP was linear in the concentration range of 0~10 mg/L, the correlation coefficient (r) was greater than 0.999 0. The relative standard deviation within and between batches was less than 10%. The recovery rate were 96.81%~101.7%. The detection limit of the method was 0.1 μg/L and the lower limit of quantification was 0.3 μg/L. Conclusion: This method has strong specificity, high sensitivity and reliable determination results. It is suitable for the rapid analysis of clinical plasma samples.
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Humanos , Cromatografia Líquida/métodos , Acetaminofen , Espectrometria de Massas em Tandem/métodos , Metanol , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Objective: To establish a method for rapid determination of bongkrekic acid (BA) in plasma by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods: In November 2020, plasma samples were extracted by methanol and acetonitrile (1∶1) and purified directly. The samples were separated by C18 column. Gradient elution was carried out with 5 mmol/L ammonium acetate water acetonitrile solution as mobile phase. Under the optimized instrument conditions, the electrospray ionization multiple reaction monitoring (MRM) mode was used, and the external standard method was used for quantitative analysis. Results: The linear relationship of BA in plasma was good in the concentration range of 2-100 μg/L, the correlation coefficient was 0.9998, the average recovery was 83.7%-112.0%, the relative standard deviation within and between batches was less than 10%, the detection limit of the method was 0.7 μg/L and the lower limit of quantification was 2.0 μg/L. Conclusion: The method is simple, rapid, accurate and sensitive, and can meet the requirements for the determination of BA in blood samples of poisoning patients.
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Humanos , Ácido Bongcréquico , Cromatografia Líquida de Alta Pressão , Extração em Fase Sólida , Espectrometria de Massas em TandemRESUMO
Objective: To explore the expulsion effect of sodium dimercaptopropanesulfonate (DMPS) on mercury in different organs of mercury poisoning and the therapeutic effect of glutathione (GSH) combined with antioxidant therapy on mercury poisoning. Methods: In February 2019, 50 SPF male SD rats were randomly divided into 5 groups, 10 rats in each group: A (saline negative control group) , B (HgCL2 positive control group) , treatment group (C: intramuscular injection of DMPS 15 mg/kg treatment, D: intramuscular injection of DMPS30 mg/kg treatment, E: intramuscular injection of DMPS 15 mg/kg and intraperitoneal injection of GSH200 mg/kg treatment) . Rats in group B, C, D and E were subcutaneously injected with mercury chloride solution (1 mg/kg) to establish a rat model of subacute mercury poisoning kidney injury. Rats in group A were subcutaneously injected with normal saline. After the establishment of the model, rats in the treatment group were injected with DMPS and GSH. Rats in group A and group B were injected with normal saline. At 21 d (treatment 7 d) and 28 d (treatment 14 d) after exposure, urine and blood samples of 5 rats in each group were collected. Blood biochemistry, urine mercury, urine microalbumin and mercury content in renal cortex, cerebral cortex and cerebellum were detected. Results: After exposure to mercury, the contents of mercury in renal cortex, cerebrum and cerebellum of rats in group B, C, D and E increased, and urine microalbumin increased. Pathology showed renal tubular injury and renal interstitial inflammation. Compared with group B, urinary mercury and renal cortex mercury in group C, D and E decreased rapidly after DMPS treatment, and there was no significant decrease in mercury levels in cerebellum and cerebral cortex of rats, accompanied by transient increase in urinary albumin after DMPS treatment (P<0.05) ; the renal interstitial inflammation in group E was improved after GSH treatment. There was a positive correlation between urinary mercury and the contents of mercury in renal cortex, cerebral cortex and cerebellum (r=0.61, 0.47, 0.48, P<0.05) . Conclusion: DMPS mercury expulsion treatment can significantly reduce the level of metal mercury in the kidney, and there is no significant change in the level of metal mercury in the cortex and cerebellum.
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Animais , Masculino , Ratos , Encéfalo/efeitos dos fármacos , Glutationa , Inflamação , Rim/efeitos dos fármacos , Nefropatias/induzido quimicamente , Cloreto de Mercúrio/uso terapêutico , Mercúrio/urina , Intoxicação por Mercúrio/tratamento farmacológico , Ratos Sprague-Dawley , Solução Salina/uso terapêutico , Unitiol/uso terapêuticoRESUMO
OBJECTIVE@#To compare the therapeutic effect of electro-nape-acupuncture (ENA) combined with hyperbaric oxygen therapy (HBOT) and single HBOT on refractory flat descending idiopathic sudden sensorineural hearing loss (ISSNHL).@*METHODS@#A total of 78 patients were randomized into an ENA combined with HBOT (ENA+HBOT) group and a HBOT group, 39 cases in each one. Patients in both groups were treated with oral extract of ginkgo biloba leaves and mecobalamin tablets. On the basis of the conventional medication treatment, HBOT was adopt in the HBOT group. On the basis of the treatment in the HBOT group, electro-nape-acupuncture was applied at Fengchi (GB 20), Gongxue (Extra), Zhongzhu (TE 3), Waiguan (TE 5) and Yifeng (TE 17), Tinggong (SI 19), Tinghui (GB 2) and the vertigo-auditory area of affected side in the ENA+HBOT group. Pulse acupuncture instrument was connected at Fengchi (GB 20) and Gongxue (Extra) for 30 min (with continuous wave, 2 Hz in frequency), the needles were retained for another 30 min after electroaupuncture. The treatment was given once a day, 6 times a week for 4 weeks in both groups. Before the treatment and 2,4 weeks into the treatment, the average auditory threshold, the scores of tinnitus handicap inventory (THI) and dizziness handicap inventory (DHI) were observed, and the therapeutic effect was evaluated in both groups.@*RESULTS@#Compared before treatment, the average auditory threshold, the scores of THI and DHI of 2,4 weeks into the treatment were decreased in both groups (0.05).@*CONCLUSION@#Electro-nape- acupuncture can improve the mean auditory threshold and the symptoms of tinnitus and dizziness in patients with refractory flat descending idiopathic sudden sensorineural hearing loss.
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Humanos , Terapia por Acupuntura , Tontura , Terapêutica , Perda Auditiva Neurossensorial , Terapêutica , Perda Auditiva Súbita , Terapêutica , Oxigenoterapia Hiperbárica , Extratos Vegetais , Usos Terapêuticos , Zumbido , Terapêutica , Resultado do Tratamento , Vitamina B 12 , Usos TerapêuticosRESUMO
Objective To study the prevalence of plasmid-mediated quinolone resistance (PMQR) genes in carbapenem-resistant Enterobacteriaceae (CRE) and its resistance mechanism.Methods Clinically isolated CRE strains in a hospital from March 2015 to March 2018 were collected, then identified and performed antimicrobial susceptibility test by VITEK2 Compact analyzer, carriage of PMQR genes qnrA, qnrB, qnrS, qepA and acc (6') Ib-cr were determined by polymerase chain reaction (PCR) and sequencing, the horizontal transfer of PMQR genes were verified by plasmid conjugation test.Results Resistance rates of carbapenem-resistant Escherichia coli and carbapenem-resistant Klebsiella pneumoniae to quinolones were 100% and 15.56%-33.33% respectively.Detection rate of acc (6') Ib-cr gene was the highest (87.72%), followed by qnrB (77.19%) and qnrS (17.54%), 2 strains (3.51%) carried qnrA gene, qepA gene was not isolated, 84.21% of strains harbored 2 or 3 PMQR genes.PMQR gene was transfected into all the 8 conjugated strains, but minimum inhibitory concentration value of quinolones didn't change significantly.Conclusion The detection rate of PMQR genes in CRE in this hospital is high, but there is a certain sensitivity to quinolones.
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<p><b>OBJECTIVE</b>To investigate whether vitamin Bphotochemical pathogen reduction technology(PRT) treatment may lead to increase white cell- and platelet- derived cytokines release from platelets during storage.</p><p><b>METHODS</b>Sixty milliliters of leukodepleted apheresis platelets were collected from 20 healthy donors, then were divided into 2 parts: one part (30 ml) remained untreated to serve as control, while the other part was treated with vitamin B-UVB photo-chemical technology as experimental group. During 7 d of storage under standard blood bank conditions, platelet coun-ting (PC), platelet distribution width (PDW), mean platelet volume (MPV), white cell-derived cytokines (IL-1β, IL-2, IL-6, IL-8, TNF-α and IFN-γ) and platelet-derived cytokines (CCL3, CCL5, TGF-β-1 and PF4), P-selectin and phosphatidyl serine (PS) were analyzed on day 1, 3, 5 and 7 of storage, respectively.</p><p><b>RESULTS</b>No signi-ficant differences were observed on PC, PDW and MPV between the experimental and control groups, respectively. The higher levels of platelet-derived cytokines were detected and reached a plateau after 5-7 days of storage, and the cyto-kines showed significant increase in experimental group compared with the control group. PS expression increased signi-ficantly in experimental group as compared with control group on day 3, 5 and 7 of storage, respectively. The accumula-tion of P-selectin was significant higher in experimental group than that in control group on day 5 and 7 of storage (P<0.05). The white cell-derived cytokines were not elevated by PRT treatment during 7 days of storage.</p><p><b>CONCLUSION</b>The PRT-treated platelets are the main source of released cytokines during storage of PRT treatment. The levels of platelet-derived cytokines reach a plateau after 5-7 days of storage, most likely due to accelerated platelet activation and apoptosis.</p>
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<p><b>OBJECTIVE</b>To investigate early serum lipid profiles in preterm infants and their relationship with neonatal respiratory distress syndrome (RDS).</p><p><b>METHODS</b>Appropriate-for-gestational-age (AGA) preterm infants were grouped according to gestational age (GA) or birth weight (BW). AGA full-term infants were randomly selected as the control group. Venous blood samples were collected within 12 hours after birth for measurement of biochemical indices, including total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG). Blood lipid levels were compared between preterm infants with and without RDS in various groups.</p><p><b>RESULTS</b>Plasma TG level rose as GA and BW increased. Plasma TG levels in the 28-30 week and 31-33 week GA groups were significantly lower than in the 34-36 week GA and control groups (P<0.01). Plasma TG levels in the ≤ 1499 g and 1500-2499 g BW groups were significantly lower than in the ≥ 2500 g BW and control groups (P<0.05), and the 1500-2499 g BW group had a significantly higher plasma TG level than the ≤ 1499 g BW group (P<0.01). There were no significant differences in plasma HDL-C, LDL-C and TC levels between all groups and between preterm infants with RDS and without RDS. In the 28-30 week GA group, the preterm infants with RDS had a significantly lower TG level than those without RDS (P<0.05); also, in the ≤ 1499 g BW group, preterm infants with RDS had a significantly lower TG level than those without RDS ( P<0.05).</p><p><b>CONCLUSIONS</b>Blood lipid levels are related to GA and BW. Low TG level may be one of the causes of RDS in preterm infants with a gestational age of 28-30 weeks and a BW of ≤ 1499 g.</p>
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Feminino , Humanos , Recém-Nascido , Masculino , Peso ao Nascer , Colesterol , Sangue , Idade Gestacional , Recém-Nascido Prematuro , Sangue , Lipídeos , Sangue , Síndrome do Desconforto Respiratório do Recém-Nascido , Sangue , Triglicerídeos , SangueRESUMO
<p><b>OBJECTIVE</b>To establish a method to determine lead and Isotope Ratios in whole blood by inductively coupled plasma mass spectrometry (ICP-MS).</p><p><b>METHODS</b>The whole blood samples was removed protein by 5% nitric acid , Online join thallium (Tl) as internal standard substance, used lyophilized bovine blood lead and cadmium standard material (GBW09139h and GBW09140h) for quality control of blood lead concentrations. Lead isotope standard substances (GBW04426) is used to determine the correction factor, lead isotope ratios will lead isotope standard reference material NIST SRM981 by icp-ms with samples for testing.</p><p><b>RESULTS</b>Optimize the detection method, detection of blood lead and lead isotope, and the method of linear range r >0.9999, GBW09139h and GBW09140h test results are within the scope of quality control. NIST SRM981 isotope determination precision RSD<1%, NIST SRM981 test results and the certificate of value close to.</p><p><b>CONCLUSION</b>The method is simple and convenient data is reliable, can meet the total lead (pb) in blood and former isotope simultaneous determination.</p>
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Animais , Bovinos , Chumbo , Sangue , Radioisótopos de Chumbo , Sangue , Espectrometria de Massas , MétodosRESUMO
<p><b>OBJECTIVE</b>To establish a method for simultaneous determination of 17 common pesticides in whole blood by solid phase extraction-gas chromatography-mass spectrometry (SPE-GC-MS).</p><p><b>METHODS</b>Whole blood samples were treated by extraction with acetonitrile, and the obtained extract was cleaned up using an Oasis HLB SPE cartridge; pesticides were separated by GC and quantitatively analyzed by MS with selected ion monitoring.</p><p><b>RESULTS</b>The concentrations of 17 pesticides in whole blood were 1.0-5.0 mg/L, and the recovery rate was 41.3-102.1%, with a relative standard deviation of less than 10%in most pesticides. The 17 pesticides showed a good linear relationship between concentration and peak area within 0.5-5.0 mg/L, with a correlation coefficient of 0.9945-0.9994. The limit of detection and limit of quantification were 0.02-0.05 mg/L and 0.05-0.09 mg/L, respectively.</p><p><b>CONCLUSION</b>With this method, 17 pesticides in whole blood can be well separated and determined. This method has high sensitivity, accuracy, and precision and can be used for identification and quantification of multiple pesticides in blood samples.</p>
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Humanos , Análise Química do Sangue , Métodos , Cromatografia Líquida , Métodos , Cromatografia Gasosa-Espectrometria de Massas , Métodos , Praguicidas , SangueRESUMO
<p><b>OBJECTIVE</b>To establish a method of determining more elements in whole blood by inductively coupled plasma mass spectrometry (ICP-MS).</p><p><b>METHODS</b>The whole blood samples were treated by 5% nitric acid to remove the proteins, then were centrifuged. Sixteen elements (Be, Al, Mn, V, Cr, Co, Ni, As, Mo, Ag, Cd, Sb, Ba,Tl, Pb and U) in the supernatant were directly measured by ICP-MS.</p><p><b>RESULTS</b>The detection limits of 16 elements were 0.01 ∼ 6.51 µg/L. The linear correlation coefficient was ≥ 0.999. The relative standard deviations were below 5 %. The recovery rates were 105% ∼ 115%. Seronorm Trace Elements SerumL-1 LOT 0903106 and GBW09139g or GBW09140g were used in the quality control, the detected results corresponded with the standard values.</p><p><b>CONCLUSION</b>ICP-MS technique is a simple, rapid, accurate and reliable method, which can be used to measure several trace elements in whole blood samples.</p>
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Humanos , Espectrometria de Massas , Métodos , Controle de Qualidade , Oligoelementos , SangueRESUMO
<p><b>OBJECTIVE</b>To establish a method to directly determine 21 elements in serum by dynamic reaction cell inductively coupled plasma mass spectrometry (DRC-ICP-MS).</p><p><b>METHODS</b>The serum samples were diluted with 1% nitric acid by 3 times. The inductively coupled plasma mass spectrometry (ICP-MS) was used simultaneously to detect the serum concentrations of 21 elements (Be, Al, Mn, V, Cr, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Ag, Cd, Sb, Ba, T1, Ph, Th, U).</p><p><b>RESULTS</b>The detection limits of all elements were between 0.001-0.0711 g/1. Standard linear correlation coefficients (r) were 50.999. Standard deviations were less than 5%,the recovery rates were 90%-114%. Standard reference materials were used for quality control standards and the analysis results conformed with the certified values.</p><p><b>CONCLUSION</b>The present method is a simple, rapid,sensitive and accurate method for detecting the serum samples.</p>
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Humanos , Limite de Detecção , Espectrometria de Massas , Métodos , Soro , Química , Análise Espectral , Métodos , Oligoelementos , SangueRESUMO
<p><b>OBJECTIVE</b>To explore correlation of seven apoptosis-related proteins (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma (ALCL).</p><p><b>METHODS</b>Using immunohistochemistry and immunofluorescence double staining methods, the expressions of these seven apoptosis-associated proteins were studied to clarify their relationship with clinical outcomes of 36 ALK+ and 25 ALK-systemic ALCL patients enrolled between 1996 and 2006. The relationship of these apoptosis-regulating proteins with NPM-ALK status was also evaluated with the tyrosine inhibitor herbimycin A (HA) in vitro by immunocytochemistry, Western blotting and flow cytometric assays.</p><p><b>RESULTS</b>The presence of Hsp90α-, MDM2-, Bax-, Cytochrome C, and Cleaved caspase3-positive tumor cells was found significantly different in ALK+ and ALK-ALCLs, which was correlated with highly favorable clinical outcome. The Bcl-2- and p53-positive tumor cells were found in groups of patients with unfavorable prognosis. Inhibition of NPM-ALK by HA could reactivate the p53 protein and subsequent apoptosis-related proteins and therefore induced apoptosis in ALK+ ALCL cells.</p><p><b>CONCLUSION</b>Our results suggest that these seven proteins might be involved in apoptosis regulation and associated with clinical outcome of ALK+ systemic ALCLs. We also reveal a dynamic chain relation that NPM-ALK regulates p53 expression and subsequent apoptosis cascade in ALK+ ALCLs.</p>
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Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Apoptose , Proteínas Reguladoras de Apoptose , Metabolismo , Benzoquinonas , Farmacologia , Biomarcadores Tumorais , Metabolismo , Western Blotting , Técnicas de Cultura de Células , Sobrevivência Celular , Intervalo Livre de Doença , Inibidores Enzimáticos , Farmacologia , Citometria de Fluxo , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Lactamas Macrocíclicas , Farmacologia , Linfoma Anaplásico de Células Grandes , Metabolismo , Patologia , Microscopia de Fluorescência , Estadiamento de Neoplasias , Prognóstico , Proteínas Tirosina Quinases , Metabolismo , Receptores Proteína Tirosina Quinases , Metabolismo , Estudos Retrospectivos , RifabutinaRESUMO
<p><b>OBJECTIVE</b>To explore the differences in the survival, adhesion and diffusion capacity between different carcinomas originating from different immunosurveillance in the same patient.</p><p><b>METHODS</b>The expressions of survivin, MMP2, TIMP1, CD44, and nm23 proteins were detected immunohistochemically in a patient with acute lymphoblastic leukemia and lymphoma after allogeneic blood stem cell transplantation.</p><p><b>RESULTS</b>Survivin, MMP2, TIMP1, CD44, and nm23 proteins were positive in acute lymphoblastic leukemia samples obtained before transplantation and negative in the lymphoma tissue occurring after the transplantation.</p><p><b>CONCLUSION</b>The expressions of survivin, MMP2, TIMP1, CD44, and nm23 proteins vary between the two carcinomas originating from different immunosurveillance in the same patient.</p>
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Adulto , Feminino , Humanos , Transplante de Células-Tronco Hematopoéticas , Receptores de Hialuronatos , Metabolismo , Proteínas Inibidoras de Apoptose , Metabolismo , Linfoma Difuso de Grandes Células B , Metabolismo , Metaloproteinase 2 da Matriz , Metabolismo , Nucleosídeo NM23 Difosfato Quinases , Metabolismo , Segunda Neoplasia Primária , Metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras , Terapêutica , Inibidor Tecidual de Metaloproteinase-1 , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To evaluate the effect of dexmedetomidine (Dex) on bispectral index (BIS) and auditory evoked potential index (AAI) during anesthesia with target controlled infusion (TCI) of propofol and remifentanyl.</p><p><b>METHODS</b>Thirty adult patients (ASA I approximate, equalsII) who were scheduled for elective thyroidectomy were monitored with BIS, AAI, ECG, blood pressure, end-tidal CO(2), and pulse oximeter before and during anesthesia. Anesthesia was induced by TCI with propofol 4 mg/L and remifentanyl 1 mu g/kg. After loss of consciousness the patients were intubated after rocuronium 0.6 mg/kg intravenous injection, remifentanyl was then infused at 0.2 microg/(kg x min)(-1) and propofol infusion (Ct) was titrated to maintain a BIS value at 50 +/- 3. At 10 min after stabilization of anesthesia the patients were randomly and double-blindly divided into 2 groups: Group D (n=15) received Dex 0.4 mu g/kg iv administered over 5 min and Group C (n=15) received equal volume of normal saline. Values of BIS, AAI, MAP, HR were recorded every 2 min within 20 min after the administration of the drugs.</p><p><b>RESULTS</b>Before anesthesia the BIS index was 90 +/- 2 in Group D and 92 +/- 2 in Group C, AAI was 81 +/- 1 in Group D and 78 +/- 1 in Group C. In anesthesia with target controlled infusion of propofol, BIS index showed a significant decrease with the i.v. administration of Dex 0.4 microg/kg, while AAI remained unchanged. In Group C, both of BIS and AAI remained unchanged after saline injection.</p><p><b>CONCLUSION</b>During propofol and remifentanyl anesthesia, after the administration of Dex, BIS value demonstrates a predominant decrease, whereas AAI shows no changes.</p>
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Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Agonistas alfa-Adrenérgicos , Androstanóis , Anestésicos Combinados , Anestésicos Intravenosos , Dexmedetomidina , Farmacologia , Método Duplo-Cego , Potenciais Evocados Auditivos , Infusões Intravenosas , Métodos , Medetomidina , Farmacologia , Monitorização Intraoperatória , Métodos , Fármacos Neuromusculares não Despolarizantes , Piperidinas , Farmacologia , Propofol , Farmacologia , TireoidectomiaRESUMO
<p><b>OBJECTIVE</b>To construct a recombinant lentivirus vector of latent membrane protein 1 (LMP1) and detect the expression of LMP1 in vitro.</p><p><b>METHODS</b>The LMP1 fragment including all the exons was amplified by PCR and inserted to the downstream of CMV promoter in the lentivirus vector pCDF. The three plasmids (packaging plasmid pFIV-34N, envelope plasmid pVSV-G and target plasmid pCDF-LMP1) were packaged into 293FT cells via liposome. The virus supernatant was harvested, concentrated and titrated. Mouse B lymphoma cell line A20 was transfected with the recombinant lentivirus vector of LMP1, and the expression of LMP1 in A20 cells was detected by RT-PCR and Western blotting.</p><p><b>RESULTS</b>DNA sequencing confirmed that the sequence of PCR-amplified LMP1 was consistent with the GenBank data. The LMP1 gene fragment was cloned into pCDF in the right direction, and the open reading frame of LMP1 was maintained. The 3 plasmids were effectively transferred into 293FT cells, which emitted green fluorescence in the cytoplasm and on the cell membrane under fluorescence microscope. The titer of the lentivirus vector reached 10(7) Tu/ml with a transfection efficiency 90% in A20 cells. LMP1 expression was detected by RT-PCR and Western blotting in transfected A20 cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector of LMP1 constructed can be effectively transfected into A20 cells, which provides a basis for exploring the role of LMP1 in the pathogenesis of lymphoma.</p>
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Humanos , Western Blotting , Linhagem Celular Tumoral , Clonagem Molecular , Vetores Genéticos , Genética , Proteínas de Fluorescência Verde , Genética , Lentivirus , Genética , Metabolismo , Linfoma de Células B , Patologia , Proteínas Recombinantes , Genética , Recombinação Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Proteínas da Matriz Viral , GenéticaRESUMO
<p><b>OBJECTIVE</b>To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 (mCD99L2) gene and observe its infection efficiency of 293FT cells.</p><p><b>METHODS</b>Four pairs of small interfering RNAs (siRNAs) targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein (EGFP) under fluorescent microscope.</p><p><b>RESULTS</b>DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1x10(7)/ml in the supernatant of the infected cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin's lymphoma.</p>
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Animais , Camundongos , Antígeno 12E7 , Antígenos CD , Genética , Sequência de Bases , DNA Complementar , Genética , Vetores Genéticos , Proteínas de Fluorescência Verde , Genética , Metabolismo , Doença de Hodgkin , Patologia , Lentivirus , Genética , Metabolismo , Dados de Sequência Molecular , Interferência de RNA , RNA Interferente Pequeno , Genética , Proteínas Recombinantes , Genética , Células Tumorais CultivadasRESUMO
<p><b>OBJECTIVE</b>To investigate the association of 3q27 chromosome rearrangement with bcl-6 gene amplification and the molecular classification, therapeutic efficacies, and clinical stages in diffuse large B cell lymphoma (DLBC).</p><p><b>METHODS</b>The newly invented cell microarray was used to detect 3q27 chromosome rearrangement and bcl-6 gene amplification in 60 cases of DLBCL by fluorescence in situ hybridization (FISH). The molecular classification of germinal center B-cell-like (GCB) and non-germinal center B-cell-like (non-GCB) was investigated by analyzing the expression of CD20, CD10, bcl-6 and MUM1 simultaneously by immunohistochemical S-P method and tissue microarray. The information of therapeutic efficacies and clinical stages was obtained by analyzing clinical cases. The relationships among the factors were analyzed by statistics.</p><p><b>RESULTS</b>In 60 cases of DLBCL, 48.3%(29/60) were GCB and 51.7%(31/60) were non-GCB. The 3q27 chromosome rearrangement and bcl-6 gene amplification were present in 15 and 22 cases respectively. In 15 cases with 3q27 rearrangement, bcl-6 protein expression was positive in 3(20.0%), which was significantly different from that in cases without 3q27 rearrangement (P=0.017). In 60 cases of DLBCL, bcl-6 gene amplification was present in 22 cases, in which 5(22.7%) were GCB and 17(77.3%) were non-GCB, which was significantly different from that in cases without bcl-6 gene amplification (P=0.003). In 36 cases undergoing the normal CHOP program treatment, bcl-6 gene amplification was present in 15 cases and the rates of the complete remission, partial remission and no change were 4(26.7%), 4(26.7%) and 7(46.7%) respectively, and again it was significantly different from that in cases without bcl-6 gene amplification (P=0.016). There were no statistical significances among bcl-6 gene, BCL-6 protein expression, and clinical stages. Cases with BCL-6 protein positive and negative expression were not correlated with therapeutic efficacies and clinical stages.</p><p><b>CONCLUSION</b>There is lower expression of BCL-6 protein in cases with bcl-6 gene fragmentation. Cases with bcl-6 gene amplification are non-GCB with worse therapeutic results and later clinical stages. There may be other genes near chromosome 3q27 associated with DLBCL prognosis.</p>
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Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Linfócitos B , Metabolismo , Aberrações Cromossômicas , Cromossomos Humanos Par 3 , Genética , Proteínas de Ligação a DNA , Genética , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , Centro Germinativo , Patologia , Hibridização in Situ Fluorescente , Linfoma Difuso de Grandes Células B , Genética , Metabolismo , Patologia , Terapêutica , Estadiamento de Neoplasias , Proteínas Proto-Oncogênicas c-bcl-6 , Análise Serial de Tecidos , Resultado do TratamentoRESUMO
The aim of this study was to investigate the expression of anaplastic lymphoma kinase (ALK) protein resulted from chromosome translocation in anaplastic large cell lymphoma (ALCL) and its relationship with the age and prognosis of patients with ALCL. The tissue microarray including 30 cases of ALCL and 2 normal control tissues were established, the expression of anaplastic lymphoma kinase (ALK) protein was detected by immunohistochemistry, the statistical analysis of detected results was carried out by SPSS software. The results showed that the ALK protein was expressed negatively in 2 cases of primary skin ALCL, but in 20 out of 28 cases of systematic ALCL the ALK protein was expressed positively and mainly located in cytoplasm and/or nucleus (71.4%). Clinically, the patients with ALK expression were younger than those without ALK expression (p < 0.05). The prognosis of patients with ALK expression was better than those without ALK expression (p < 0.05). It is concluded that there is a high incidence of ALK expression in ALCL, especially in younger group. ALK expression may be an useful and independent marker for the differential diagnosis and prognosis evaluation of ALCL.
Assuntos
Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Cromossomos Humanos Par 1 , Cromossomos Humanos Par 2 , Linfoma Anaplásico de Células Grandes , Genética , Prognóstico , Proteínas Tirosina Quinases , Genética , Metabolismo , Receptores Proteína Tirosina Quinases , Translocação GenéticaRESUMO
<p><b>OBJECTIVE</b>To compare the efficacy of nuclear microarray combined with fluorescence in situ hybridization (FISH) and immunohistochemistry in detecting ALK gene translocation and ALK fusion protein in anaplastic large cell lymphoma (ALCL).</p><p><b>METHODS</b>ALK gene translocation and ALK fusion protein in 17 paraffin-embedded ALCL specimens were detected using nuclear microarray combined with FISH and immunohistochemical straining, respectively.</p><p><b>RESULTS</b>The expression of ALK fusion protein was detected immunohistochemically with ALK antibody in 8 of the 17 specimens of systemic ALCL, including 4 with both nuclear and cytoplasmic positivity and 4 with only cytoplasmic positivity. Dual-color FISH identified 6 positive specimens, including the 4 specimens with both nuclear and cytoplasmic positivity as identified immunohistochemically, and 2 with immunohistochemical cytoplasmic positivity. FISH yielded negative results for the 2 specimens with immunohistochemical cytoplasmic positivity.</p><p><b>CONCLUSION</b>Nuclear microarray combined with FISH eliminated the cytoplasmic interference of the results of conventional FISH and provides a high-throughput platform for clinical detection with greater specificity than immunohistochemistry.</p>
Assuntos
Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Métodos , Linfoma Anaplásico de Células Grandes , Genética , Patologia , Análise em Microsséries , Métodos , Inclusão em Parafina , Proteínas Tirosina Quinases , Genética , Receptores Proteína Tirosina Quinases , Reprodutibilidade dos Testes , Translocação GenéticaRESUMO
Apesar da epilepsia ser a condição neurológica grave mais comum existente no mundo, crenças e comportamentos inadequados ainda persistem. Para mudar esta perspectiva, o Projeto Demonstrativo da Campanha Global "Epilepsia fora das sombras" permitiu o engajamento das associações de epilepsia em nosso país, fortalecendo o Movimento Nacional de Epilepsia. Uma das atividades deste movimento é a realização da Semana Nacional de Conscientização de Epilepsia, que acontece todos os anos na semana do dia 09 de setembro, com o objetivo de conscientizar a sociedade sobre a epilepsia. A ASPE realiza esta Semana desde 2003 e os resultados trazem importante fortalecimento ao movimento nacional para tirar a epilepsia das sombras e melhorar a qualidade de vida das pessoas com epilepsia e suas famílias.
Epilepsy is a common neurological condition; however, it is still very frequent to observe myths and inadequate behaviors regarding epilepsy in our society. The National Demonstration Project "Epilepsy out of the shadows" brought important changes in our society through a national movement of epilepsy carried out by lay associations. The National Week of Epilepsy, on the September 9th is one the major national movements with purpose to promote epilepsy awareness within our society about epilepsy. ASPE, a non-governmental organization, participates actively in the movement since its origin in 2003. We believe that the results contribute to reinforce the national movement to bring epilepsy out of the shadows, diminishing the associated stigma and improving the quality of life of people with epilepsy and their families.