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OBJECTIVES@#To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS).@*METHODS@#The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (Vacetonitrile∶Vmethanol=2∶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring.@*RESULTS@#The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 μg/mL and 252.14 ng/mL, respectively.@*CONCLUSIONS@#This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.
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Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem , Metanol , Carbamazepina/análise , Benzodiazepinas/análise , Solventes , Cromatografia Líquida de Alta Pressão , Extração em Fase SólidaRESUMO
Objective: To analyze family-based haplotype frequencies of HLA-A, -B, -C, -DRB1 and -DQB1 genes and their clinical significance. Methods: The data of HLA genotyping in 3568 families undergoing related haploidentical transplantation between 2012 and 2017 at the First Affiliated Hospital of Soochow University were retrospectively evaluated. The HLA genotyping was performed by PCR amplification with sequence-based typing (PCR-SBT) and sequence-specific oligonucleotide probe (PCR-SSOP) methods. The family genetic analysis and haplotype frequencies were also investigated. Results: All the families were divided into 3 groups, including group1 of 1 422 entire families; group2 of 1 310 patients and either of their parents or one of their children; group3 of 836 patients and their HLA≥5/10 matched sibling donors. In the haplotypes with frequencies greater than 0.1% in group1+ group2, the frequency of A*11∶01-B*40∶01-C*03∶04-DRB1*11∶01-DQB1*03∶01, A*02∶07-B*51∶01-C*14∶02-DRB1*09:01-DQB1*03∶03 were significantly different between group1 and group2 (P=0.029, 0.033) . The frequency of A*11∶01-B*46∶01-C*01∶02∶01G-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group3 (P=0.035) . The frequency of A*02∶01-B*40∶01-C*07∶02-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group2 (P=0.034) , or group1 and group3 (P=0.034) . The frequency of A*24∶02-B*13∶01-C*03∶04-DRB1*12∶02-DQB1*03:01 was significantly different between group2 and group3 (P=0.046) . Conclusion: In this study, we summarize the prevalence of haplotype frequencies in terms of HLA-A, -B, -C, -DRB1 and-DQB1. Based on the database of family haplotype analysis, patients and donor candidates are sorted with matched HLA genotype while unmatched HLA haplotype. Even in patients without entire family information, HLA haplotype analysis assists in choosing the optimal related or unrelated donors.
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Criança , Humanos , Alelos , Frequência do Gene , Antígenos HLA-A , Antígenos HLA-B , Antígenos HLA-C , Cadeias beta de HLA-DQ , Cadeias HLA-DRB1 , Haplótipos , Estudos RetrospectivosRESUMO
@#In order to provide enlightening experience for the establishment of reasonable and diversified orphan drugs into the reimbursement system in China, the official websites of the National Institute for Health and Care Excellence(NICE)and the National Health Service(NHS)in the UK were inspected, collecting and summarizing the relevant documents, on the inclusion of orphan drugs into the reimbursement system. Relevant literatures were analyzed with theoretical studies on the accessibility of medicines for patients with rare diseases. In the NHS system, the inclusion of orphan drugs into the reimbursement system in the UK can be achieved mainly through seven routes, with three routes that are evaluated by NICE(MTA, STA and HST)and four are directly managed by the NHS(specialized commissioning, CDF, IFRs, CtE). Through the analysis of the inclusion of orphan drugs into the reimbursement system and the various problems in the UK, we have found some enlightening experience for the establishment of the reimbursement system for orphan drugs in China.
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<p><b>OBJECTIVE</b>To investigate the protective effects of oxymatrine on chronic heart failure induced by isoproterenol (ISO) and to observe its effects on ADMA metabolism pathway in ISO-induced chronic heart failure in rats.</p><p><b>METHOD</b>Male Sprague-Dawley rats were given oxymatrine (100,50 mg kg-1) orally for 14 days. Heart failure was induced in rats by subcutaneous injection of isoproterenol (5 mg kg-1 d-1 ) at the 8th day for 1 week. Serum parameters, haemodynamic parameters, Heart weight, and histopathological variables were analysed. Expression of protein levels were measured by Western blot.</p><p><b>RESULT</b>Oxymatrine (100,50 mg kg-1) significantly attenuated serum content of cTn I, improved left ventricle systolic and diastolic function and left ventricular remodeling, reduced the ISO-induced myocardial pathological changes compared with ISO group. In addition, oxymatrine (100,50 mg kg-1) significantly reduced serum level of ADMA (P <0. 01), normalize the reduced dimethylarginine dimethylaminohydrolase 2 (DDAH2) expression (P <0. 01) , but had no effect on the isoproterenol-induced upregulated protein arginine methyltransferases 1 expression.</p><p><b>CONCLUSION</b>Oxymatrine could ameliorate the experimental ventricular remodeling in ISO-induced chronic heart failure in rats and the mechanism involved in reducing serum content of ADMA and increased DDAH2 expression.</p>
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Animais , Masculino , Ratos , Alcaloides , Farmacologia , Usos Terapêuticos , Amidoidrolases , Metabolismo , Arginina , Sangue , Metabolismo , Doença Crônica , Regulação Enzimológica da Expressão Gênica , Insuficiência Cardíaca , Tratamento Farmacológico , Metabolismo , Patologia , Hemodinâmica , Isoproterenol , Tamanho do Órgão , Quinolizinas , Farmacologia , Usos Terapêuticos , Ratos Sprague-Dawley , Troponina I , MetabolismoRESUMO
This study was purposed to investigate the clinical value of HLA matching(low and high resolution) and its effect on outcome of the patients received umbilical cord blood transplantation(UCBT). Sequence-specific oligonucleotide probe (SSOP) , sequence-based typing (SBT) and sequence-specific primers(SSP) were used to perform high resolution HLA matching for HLA-A, -B, -Cw, -DRB1, -DQB1 and low resolution for HLA-A, B, DRB1 among 34 patients with hematologic malignancies who received unrelated UCB transplantation and grafts. The effects of HLA matching (low or high resolution ) on leading engraftment, hematopoietic reconstitution, graft-versus-host disease (GVHD) and infection after UCB transplantation were analyzed by comparison. The results showed that the median of total nucleated cells (TNC) of transplanted cord blood was 6.0×10(7)/kg, The time of neutrophil recovery was significantly shortened when more than 5×10(7)/kg TNC were transplanted (P < 0.05). The HLA-(6-10)/10 group of high resolution HLA matching was better than the HLA (4-5)/10 group in the respect of leading engraftment, the time of platelet recovery and the rate of acute GVHD (P < 0.05). In contrast, HLA-I+II locus, HLA-DRB1 or HLA-DQB1 locus mismatch could prolong the platelet engraftment time (P < 0.05). There was statistical difference in the time of platelet recovery, the rate of acute GVHD between the HLA (5-6)/6 group of low resolution HLA matching and the HLA (3-4)/6 group after UCB transplantation (P < 0.05), but the mismatch locus of HLA with low resolution did not correlate with the time of platelet recovery (P > 0.05). It is concluded that the high resolution HLA matching between patients received unrelated UCB transplantation and grafts may contribute to select the better UCB, that has important clinical value to promote hematopoietic reconstitution and to reduce the complications after UCB transplantation.
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Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Adulto Jovem , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Métodos , Neoplasias Hematológicas , Terapêutica , Teste de Histocompatibilidade , MétodosRESUMO
<p><b>BACKGROUND</b>Humoral immunity is an important factor for long-term survival of renal allograft. Here we performed a four-year follow-up to explore the clinical significance of monitoring anti-human leukocyte antigens (HLA) and anti-major histocompatibility complex class I-related chain A (MICA) antibody expression after kidney transplantation.</p><p><b>METHODS</b>We obtained serial serum samples from 84 kidney transplant patients over a four-year period. All patients were followed up at least 6 months after transplantation and had at least two follow-up points. Anti-HLA and anti-MICA antibody titres and serum creatinine (SCr) levels were evaluated at each follow-up. Patients were divided into 4 groups: HLA(+) MICA(-), HLA(-)MICA(+), HLA(+)MICA(+) and HLA(-)MICA(-). The impact of post-transplant antibody level on kidney allograft function was evaluated.</p><p><b>RESULTS</b>Antibodies were detected in 38.1% (32/84) of the renal allograft recipients. HLA, MICA and HLA+MICA expression was observed in 18.89%, 14.44% and 5.93% of the recipients respectively. The most frequent anti-HLA and anti-MICA specific antibodies identified were A11, A24, A29, A32, A33, A80; B7, B13, B37; DR17, DR12, DR18, DR52, DR53, DR1, DR4, DR9, DR51; DQ7, DQ4, DQ8, DQ2, DQ9, DQ5, DQ6 and MICA02, MICA18, MICA19, MICA07, MICA27. As the time after transplantation elapsed, more recipients developed de novo antibody expression. Total 11.91% (10/84) of the recipients had de novo antibody expression during the follow up. The average level of SCr and the percentage of recipients with abnormal allograft function were significantly higher in recipients with anti-HLA and/or anti-MICA antibody expression than those without. The appearance of anti-HLA and anti-MICA antibody expression always preceded the increase in SCr value.</p><p><b>CONCLUSIONS</b>Anti-HLA and anti-MICA antibody expression has predictive value for early and late allograft dysfunction. The presence of donor specific antibody is detrimental to graft function and graft survival.</p>
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Feminino , Humanos , Masculino , Seguimentos , Sobrevivência de Enxerto , Antígenos HLA , Alergia e Imunologia , Antígenos de Histocompatibilidade Classe I , Alergia e Imunologia , Isoanticorpos , Transplante de Rim , Antígenos de Histocompatibilidade MenorRESUMO
<p><b>OBJECTIVE</b>To find out the distributed characteristics of KIR2DL1 alleles frequencies and the recognition HLA-C ligand in the Chinese Han population.</p><p><b>METHODS</b>The 111 patients and 116 donors from CMDP were performed the KIR2DL1 high-resolution typing and KIR genotyping using sequence-based testing (SBT) and PCR-SSP methods.</p><p><b>RESULTS</b>A total of 224 individuals with KIR2DL1 locus was predominantly observed and accounted for 98.68% (224/227). There were 3 different KIR2DL1 alleles, including KIR2DL1*00302, *00201 and *00401 alleles polymorphism. The most common phenotype observed were KIR2DL1*00302 (84.82%, 380/448), KIR2DL1*00201 (12.05%, 54/448) and KIR2DL1*00401(3.13%,14/448), present at allele genotype frequencies of 61.04%,6.22% and 1.58% respectively. The allele homozygotic types of KIR2DL1*00302 and KIR2DL1*00302 were the most frequent in 6 KIR2DL1 allele by high resolution typing. The allele heterozygous types of KIR2DL1*00302 and KIR2DL1*00401 presented statistically different in haplotypes A/A and B/x (P=0.001), and KIR2DL1*00401 lacked of all A/A haplotype. The KIR2DL1*00302 and KIR2DL1*00201 allele had significant positive associations between different KIR pairs of KIR2DS1, KIR2DL3, KIR2DS4 and KIR3DL1/S1 using linkage disequilibrium analysis (P<0.01), respectively. In the receptor-ligand of KIR/HLA model after allo-HSCT, KIR2DL1*00302 alleles correlated with their HLA-C2 group ligands. KIR2DL1*00302 and HLA-C*06:02 was the most common combination ligand model, but KIR2DL1*00302 and HLA-C*01:02 was the most frequent mismatch ligand model with the development of NK cell-induced alloreactivity, meanwhile there was statistically significant difference of frequency distribution (P<0.05).</p><p><b>CONCLUSION</b>The KIR2DL1*00302 was the most frequent allele in Chinese Han population. The KIR2DL1 high resolution typing would be beneficial for predicting donor NK cells all activity after hematopoietic stem cell transplantation and selecting suitable donors.</p>
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Humanos , Alelos , Povo Asiático , Genética , Frequência do Gene , Genótipo , Antígenos HLA-C , Genética , Haplótipos , Teste de Histocompatibilidade , Ligantes , Polimorfismo Genético , Receptores KIR2DL1 , GenéticaRESUMO
<p><b>OBJECTIVE</b>To study the impact of various human leukocyte antigen (HLA) high resolution typing mismatching of donor-recipient pairs on prognosis of unrelated donor hematopoietic stem cell transplantation.</p><p><b>METHODS</b>835 donor-recipient pairs of CMDP data from 2005 to 2010 were analyzed retrospectively. HLA-A, B, C, DRB1 and DQB1 typing were performed using SBT, SSOP and SSP methods. The diseases involved in acute myeloid leukemia (AML) (n = 288), acute lymphoid leukemia (ALL) (n = 227), chronic myeloid leukemia (CML) (n = 187), myelodysplastic syndrome (MDS) (n = 52), non-hodgkin's lymphoma(NHL) (n = 25), aplastic anemia(AA) (n = 42) and thalassemia (n = 14). Of 835 donor-recipient pairs, 362 were completely matched, 159 had a mismatch for a single allele, 125 had a mismatch for a single antigen, 95 had mismatched for both single allele and single antigen, 29 were mismatched at double allele, 20 at double antigen, 45 at multiple allele and antigen. The follow-up assessment was completed before March 2011.</p><p><b>RESULTS</b>HLA-matched pairs had higher overall survival (OS) than HLA-mismatched pairs (79.83% vs 73.15%), but there was no statistically significant differences (P > 0.05). HLA mismatch for a single allele plus a single antigen was a significantly risk factor for OS, disease free survival (DFS) and transplant-related mortality (TRM). The OS from high to low in different diseases were thalassemia, AA, CML, MDS, AML, NHL, and ALL. OS of HLA locus mismatch were DRB1 (94.4%), DQB1 (83.3%), B (75%), A (74.4%) and C (71.4%), respectively. OS of single allele mismatch at HLA locus from high to low were DRB1, C, A, B and DQB1.HLA-A, B, C locus mismatch were statistically significantly associated with lower OS and grade II-IV acute GVHD compared with HLA-matched pairs (P < 0.05). The donor-recipient pairs with HLA-B*15:01/B*15:05, DRB1*12:01/DRB1*12:02, C*04:01/C*03:04, DQB1*03:02/DQB1*03:03 alleles mismatch were given priority. But the donor-recipient pairs with HLA-B*39:01/B*39:05, C*15:02/C*14:02, C*08:01/C*03:04, C*07:02/C*15:02 alleles mismatch were risk factors for influence of OS and aGVHD.</p><p><b>CONCLUSION</b>The high resolution typing for HLA-A, B, C, DRB1, DQB1 can be identified nonpermissive mismatch, which is beneficial for the selection of a suitable donor improves survival on unrelated donor HSCT.</p>
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Humanos , Antígenos HLA , Genética , Alergia e Imunologia , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Leucemia Mieloide Aguda , Alergia e Imunologia , Cirurgia Geral , Linfoma não Hodgkin , Alergia e Imunologia , Cirurgia Geral , Síndromes Mielodisplásicas , Alergia e Imunologia , Cirurgia Geral , Prognóstico , Estudos Retrospectivos , Doadores não RelacionadosRESUMO
<p><b>OBJECTIVE</b>To analyze the allele frequencies and polymorphism of human leukocyte antigens (HLA) -A, B, Cw, DRB1 and DQB1 between donors-recipients on high-resolution typing; and to analyze the matching and mismatching proportion between donors and recipients.</p><p><b>METHODS</b>HLA high-resolution types were determined by sequence based typing (SBT), sequence specific oligonucleotide probe (SSOP) and sequence specific primer (SSP) on 2540 unrelated Chinese Han individuals including 1168 recipients and 1372 donors, then statistical analyses were carried out.</p><p><b>RESULTS</b>Forty-four HLA-A alleles were detected, and among them the frequencies of A*1101, A*2402, A*0201, A*0207, A*3303, A*0206 and A*3001 exceeded 0.05, and accounted for 80.4%. Eighty-one HLA-B alleles were detected, and the frequencies of B*4001, B*4601, B*5801, B*1302 and B*5101 exceeded 0.05, and accounted for 43.0% of total. There were 44 HLA-Cw alleles, among them the frequencies of Cw*0702, Cw*0102, Cw*0304, Cw*0801, Cw*0602, Cw*0303, Cw*0302 and Cw*0401 exceeded 0.05, and were 80.3% of total. There were 61 HLA-DRB1 alleles, the frequencies of DRB1*0901, DRB1*1501, DRB1*1202, DRB1*0803, DRB1*0701, DRB1*0405, DRB1*0301 and DRB1*1101 exceeded 0.05, and were 70.1% of total. Finally, 22 HLA-DQB1 alleles were detected, the frequencies of DQB1*0301, DQB1*0303, DQB1*0601, DQB1*0602, DQB1*0202, DQB1*0302, DQB1*0401, DQB1*0502 and DQB1*0201 exceeded 0.05, and they were 87.4% of total. All the five loci were of heterozygote deficiency. The HLA-A, B and DRB1 loci conformed to Hardy-Weinberg equilibrium (HWE) (P > 0.05); but HLA-Cw and HLA-DQB1 loci did not (P < 0.05). Except several particular genotypes, all the five loci conformed to HWE. After comparing data between donors and recipients, only 22.4% of recipients found HLA matched donors (10/10); 24.6% of recipients found single HLA allele mismatched donors (9/10); 26.3% of recipients had two HLA alleles mismatched donors (8/10).</p><p><b>CONCLUSION</b>The characteristics of allele frequencies and polymorphism of HLA-A, B, Cw, DRB1 and DQB1 on high-resolution typing in Chinese Han population is valuable for donor searching in unrelated hematopoietic stem cell transplantation, and it provides genetic basis for donor registry and usage of donor resource for Chinese Marrow Donor Program.</p>
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Humanos , China , Etnologia , Frequência do Gene , Genética Populacional , Antígenos HLA-A , Genética , Antígenos HLA-B , Genética , Antígenos HLA-C , Genética , Antígenos HLA-D , Genética , Antígenos HLA-DQ , Genética , Cadeias beta de HLA-DQ , Antígenos HLA-DR , Genética , Cadeias HLA-DRB1 , Transplante de Células-Tronco Hematopoéticas , Antígenos de Histocompatibilidade Classe I , Genética , Teste de Histocompatibilidade , Doadores de TecidosRESUMO
<p><b>OBJECTIVE</b>To detect the frequencies of KIR2DS4 alleles in Chinese Han population and to study the impact of KR2DS4 alleles on clinical outcomes of HLA identical unrelated allo-HSCT.</p><p><b>METHODS</b>A Sequence-Based Testing (SBT) and TOPO TA cloning system for identifying and distinguishing alleles of the KIR2DS4 gene were established. A total of 150 Chinese-Han individuals, including 75 leukemia patients who received allo-HSCT and their HLA high-resolution typing identical unrelated donors (URD) were entered this study. The patients underwent transplantation for CML (n = 24), AML (n = 19), ALL (n = 29) and other malignancies (n = 3).</p><p><b>RESULTS</b>The majority (139) of the 150 samples (92.7%) were positive for KIR2DS4. Sequencing of the whole length coding region of this gene identified four of the 12 known KIR2DS4 alleles, KIR2DS4*00101, *003, *004, and *007. 2DS4*00101 was the most frequent, being found in 109 of the 139 individuals (78.4%). The ratio of deleted to non-deleted versions of KIR2DS4 was approximately 1:2. Three novel KIR2DS4 alleles were identified. Transplantations from KIR haplotype B/x donors showed significantly higher overall survival rates than those from KIR haplotype A/A donors [RR 3.1 (95%CI 1.1 - 8.6), P = 0.007]. There was a lower overall survival rates in recipients when their donors carried two 2DS4 full-length allele (2DS4*001) than those carried less (0 or 1) 2DS4*001 allele (P = 0.031). In the haplotype A/A group, a higher risk of acute GVHD (aGVHD) [RR 9.0 (95%CI 1.2 - 66.9), P = 0.01], especially grade III-IV aGVHD (P = 0.006), was seen when the donor was homozygous for the full-length KIR2DS4*00101 allele.</p><p><b>CONCLUSION</b>The development and application of the described SBT 2DS4 allele typing method highlights the diverse nature of the KIR gene family and displays the existence of KIR polymorphism that remains uncharacterized. Our findings suggest that KIR typing for 2DS4 be beneficial for selecting suitable donors.</p>
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Humanos , Alelos , Doença Enxerto-Hospedeiro , Genética , Haplótipos , Transplante de Células-Tronco Hematopoéticas , Receptores KIR , GenéticaRESUMO
<p><b>OBJECTIVE</b>To explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>From January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.</p><p><b>RESULTS</b>Of 48 patients, 15 were transplanted from unrelated donors with an antigen mismatch for HLA Cw and 33 patient-donor pairs were matched for HLA-Cw. The CMV reaction rate was 66.7% for HLA-Cw mismatch group and 48.5% for HLA-Cw match group (chi(2) = 1.39, P = 0.2375). Thirty-seven donor-patients pairs belonged to group C1 and 11 to group C2, and CMV reaction rate was 64.9% in group C1 and 18.2% in group C2 (chi(2) = 18.13, P < 0.0001). Twenty-six patients received graft from KIR haplotype A (group A donor) and 22 from KIR haplotype B donors (group B donor) and CMV reaction rate was 57.7% in group A donor and 50.0% in group B donor (chi(2) = 0.28, P = 0.5941). The number of donor activating KIRs (aKIRs) was less than that of recipient aKIRs in 34 patient-donor pairs in which the CMV reaction rate was 70.6%, and the number of donor aKIRs was more than that of recipient aKIRs in 14 patient-donor pairs in which the CMV reactivation was 14.3%. There was a significan difference between the two group (chi(2) = 12.44, P = 0.0004).</p><p><b>CONCLUSION</b>KIR and HLA-Cw genotypes influence the rate of CMV reactivation following non-T cell deleted unrelated donor hematopoietic cell transplantation.</p>
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Humanos , Genótipo , Antígenos HLA-C , Genética , Haplótipos , Transplante de Células-Tronco Hematopoéticas , Receptores KIR , GenéticaRESUMO
<p><b>OBJECTIVE</b>To establish a real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) for quantitative detection of the common molecular markers that have affirmative clinical significance in the acute and chronic leukemia patients, and evaluate its significance in diagnosing leukemias and monitoring minimal residual disease (MRD).</p><p><b>METHODS</b>Primers and TaqMan probes were designed for detecting various fusion transcripts and normal abl gene was used as the internal control. The expression level of fusion transcripts in 202 newly diagnosed leukemias were determined.</p><p><b>RESULTS</b>In absolute quantity, expression level of the fusion transcripts in various leukemias was b3a2 (b2a2) 47614.63, e1a2 98847.53, AML1-ETO 300029.51, PML-RAR alpha 25506.28, respectively, while in relative quantity to abl, the levels were 1.05, 0.91, 5.33 and 0.55, respectively.</p><p><b>CONCLUSION</b>The relative quantification of gene expression level by using RQ-RT-PCR to abl control gene is more accurate and direct viewing. Different levels of transcription of corresponding fusion genes are found in various subtypes of leukemias at diagnosis, among which the level of AML1-ETO was higher and PML-RAR alpha lower.</p>
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Humanos , Biomarcadores Tumorais , Genética , Metabolismo , Subunidade alfa 2 de Fator de Ligação ao Core , Genética , Metabolismo , Proteínas de Fusão bcr-abl , Genética , Metabolismo , Leucemia , Diagnóstico , Genética , Metabolismo , Proteínas de Fusão Oncogênica , Genética , Metabolismo , Proteína 1 Parceira de Translocação de RUNX1 , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição GênicaRESUMO
<p><b>OBJECTIVE</b>To study the biological function of killer cell immunoglobulin-like receptor (KIR) and the role of donor inhibitory KIR and recipient genetic background in HLA matched unrelated hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>HLA genotype of 51 patients (ALL 18 cases, CML 15 cases, AML 10 cases and others 8 cases) and their respective matched unrelated donors from Database of China Marrow Registration was determined by polymerase chain reaction sequence oligonucleotide probes (PCR-SSOP) and sequence specific primers (PCR-SSP). The KIR genotype was determined by PCR-SSP.</p><p><b>RESULTS</b>All the patients and the donors expressed KIR2DL1, KIR2DL2/L3, KIR2DL4, KIR3DL2 and KIR3DL3. 96.7% individuals expressed KIR3DL1. Among them, 21.57% of KIR was completely identical, while 78.43% was not. Of the non-identical KIRs, 25.49% were the recipient's KIR genotype containing the donor's ones, and 27.45% was the donor's containing the recipient's. 74.62% of donor's KIR2DL1 lacked recipient's C2 ligand, 5.91% of donor's KIR2DL2/L3 lacked recipient's C1 ligand, 19.74% of donor's KIR3DL1 lacked recipient's Bw4 ligand and 54.91% of donor's KIR3DL2 lacked recipient's A3, A11 ligand.</p><p><b>CONCLUSION</b>KIR genotype and HLA class I antigen are inherited independently. KIR2DLI and KIR3DL2 of donors may cause alloreactivity of NK cell. The mismatch of KIR/HLA in donor-recipient plays a very important role in matched unrelated allo-HSCT. The outcome of HSCT can be better predicted by the model of the presence of KIRs on the donor' sNK cells and the absence of corresponding KIR ligand in the recipient's HLA.</p>