RESUMO
OBJECTIVE: To investigate the relationship between microRNA-34b/c single nucleotide polymorphism (SNP) rs4938723 and the risk of male infertility. METHODS: This case-control study included 553 males aged 19ï¼40 (29.42 ± 5.09) years with idiopathic infertility, 153 with azoospermia and 400 with oligoasthenospermia, and another 332 normal fertile men aged 19ï¼40 (28.5 4 ± 4.63) years as controls. We collected the clinical data and peripheral blood samples from the subjects, genotyped microRNA-34b/c rs4938723 by Sequenom MassARRAY, and analyzed the relationship between the genotypes of microRNA-34b/c rs4938723 and the risk of male infertility using the logistic regression model. RESULTS: Statistically significant differences were observed between the infertility patients and normal controls in sperm concentration (ï¼»18.71 ± 15.19ï¼½ vs ï¼»79.91 ± 43.96ï¼½ × 106/ml, P < 0.01), the percentage of progressively motile sperm (ï¼»13.27 ± 24.52ï¼½% vs ï¼»42.82 ± 8.86ï¼½%, P < 0.01) and the level of follicle stimulating hormone (ï¼»16.09 ± 17.37ï¼½ vs ï¼»12.20 ± 4.73ï¼½ IU/L, P < 0.01). Compared with the TT genotype, the TC and CC genotypes showed no correlation with male infertility, nor did the genetic locus in the subgroup analysis. CONCLUSIONS: No correlation was found between microRNA-34b/c SNP rs4938723 and male infertility, which, however, needs to be further verified by larger-sized samples.
Assuntos
Infertilidade Masculina , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único , Adulto , Estudos de Casos e Controles , Predisposição Genética para Doença , Genótipo , Humanos , Infertilidade Masculina/genética , Masculino , Adulto JovemRESUMO
OBJECTIVE: To investigate the mutation of the DPY19L2 gene in patients with globozoospermia. METHODS: We collected the clinical data and peripheral blood from 2 patients with globozoospermia and screened for mutation of the DPY19L2 gene by PCR amplification and DNA sequencing technology. RESULTS: The sperm from the 2 globozoospermia patients were round morphologically under the light microscope, with deeply stained nuclei but no acrosome. Electron microscopy showed the sperm with a large round head but no acrosomal structure, the nuclei enveloped by a single layer of membrane and the cytoplasm dispersed. PCR amplification revealed homozygous deletion of Exon 5, Exon6 and Exon15 in the DPY19L2 gene in both the patients. CONCLUSIONS: This study proved that the homozygous mutation of DPY19L2 could lead to globozoospermia, which has an important significance for researches on the molecular mechanisms and gene diagnosis of the disease as well as for clinicians in genetic counseling and treatment.
Assuntos
Proteínas de Membrana/genética , Teratozoospermia , Homozigoto , Humanos , Masculino , Mutação , Deleção de Sequência , Espermatozoides , Teratozoospermia/genéticaRESUMO
OBJECTIVE: Detection of azoospermia factor (AZF) microdeletions on the Y chromosome is one of the auxiliary strategies recognized at home and abroad for the examination of male infertility. Traditional PCR gel electrophoresis fails to meet the clinical needs due to its shortcomings. The purpose of this study was to explore the feasibility of multiplex fluorescence PCR in the detection of AZF microdeletions. METHODS: We collected samples of Y chromosomal AZF microdeletions from 238 patients with azoospermia or oligozoospermia and 62 normal males, identified the 14 short tandem repeat (STR) loci in the AZF region of the Y chromosome by multiplex PCR gel electrophoresis and multiplex fluorescence PCR, and analyzed the consistency in the results of the two methods by Kappa test. RESULTS: There was a perfect consistency between multiplex PCR gel electrophoresis and multiplex fluorescence PCR in the detection rate of the STR loci in the 300 samples. Kappa test showed both P and Kappa values to be 1 for the 6 loci in the AZFa, AZFb and AZFc regions of the Y chromosome, with no statistically significant difference between the two methods. CONCLUSIONS: Multiplex fluorescence PCR can save a lot of time, reduce workload and improve laboratory efficiency and therefore is preferable to multiplex PCR gel electrophoresis in detecting Y chromosome microdeletions.
Assuntos
Azoospermia , Deleção Cromossômica , Cromossomos Humanos Y , Infertilidade Masculina , Reação em Cadeia da Polimerase Multiplex , Azoospermia/genética , Cromossomos Humanos Y/genética , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/genética , Masculino , Oligospermia/genéticaRESUMO
Objective: To investigate the correlation between the single nucleotide polymorphism (SNP) rs662 of the paraoxonase 1 gene (PON1) and the risk of male infertility. METHODS: This case-control study included 403 male idiopathic infertility patients aged 29.00 ± 4.48 years in the case group and 329 normal fertile men aged 28.28 ± 4.08 years as healthy controls. We obtained DNA from the peripheral venous blood of the subjects, genotyped the SNP rs662 of PON1 by Sequenom MassArray, and analyzed the association between different genotypes of PON1 rs662 and male infertility using the logistic regression model. RESULTS: Compared with the normal controls, the infertility patients showed a significantly increased level of follicle-stimulating hormone (FSH) (ï¼»16.30 ± 17.76ï¼½ vs ï¼»4.72 ± 2.51ï¼½ U/L, P < 0.01) but a decreased percentage of progressively motile sperm (PMS) (ï¼»7.40 ± 14.17ï¼½ % vs ï¼»41.93 ± 9.06ï¼½ %, P < 0.01) and sperm concentration (ï¼»2.74 ± 3.64ï¼½ vs ï¼»75.83 ± 63.66ï¼½ ×106/ml, P < 0.01). Statistically significant differences were not found in the other parameters between the two groups of subjects, nor in the correlation of male infertility with the heterozygous genotype GA versus the wild homozygous genotype GG (OR = 0.98, 95% CI: 0.63ï¼1.53, P = 0.923) or the homozygous genotype AA versus the wild homozygous genotype GG (OR = 0.87, 95% CI: 0.56ï¼1.34, P = 0.525). CONCLUSIONS: The SNP rs662 of PON1 was not correlated with male infertility, which, however, needs to be confirmed by further studies with larger samples from a larger area.
Assuntos
Arildialquilfosfatase/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Predisposição Genética para Doença , Genótipo , Heterozigoto , Homozigoto , Humanos , Infertilidade Masculina/sangue , Modelos Logísticos , Masculino , Contagem de Espermatozoides , Adulto JovemRESUMO
OBJECTIVE: To investigate the correlation of the single nucleotide polymorphism (SNP) rs1042522 of the tumor protein p53 (TP53) gene with the risk of male infertility. METHODS: This caseîcontrol study included 380 male patients with idiopathic infertility and 398 normal fertile men as controls from the Nanjing area. We genotyped the SNP rs1042522 of the TP53 gene by Sequence Mass Array and analyzed the correlation of the SNP with male infertility using the logistic regression model. RESULTS: Compared with the normal controls, the patients with idiopathic infertility showed significantly decreased sperm concentration (ï¼»77.34±49.24ï¼½ vs ï¼»13.13±24.96ï¼½ ×106/ml), percentage of progressively motile sperm (ï¼»42.55±9.57ï¼½ vs ï¼»10.38±5.57ï¼½%), serum testosterone level (ï¼»14.07±5.36ï¼½ vs ï¼»11.89±4.50ï¼½ nmol/L), and follicleîstimulating hormone level (ï¼»16.80±18.20ï¼½ vs ï¼»4.55±7.17ï¼½ U/L) (P < 0.05) but no statistically significant differences in other parameters. No correlation was observed between the SNP frequencies and male infertility and similar results were found in the subgroups of the cases. CONCLUSIONS: SNP rs1042522 of the TP53 gene is not significantly correlated with the risk of male infertility.
Assuntos
Genes p53/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Contagem de Espermatozoides , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Infertilidade Masculina/sangue , Modelos Logísticos , Masculino , Motilidade dos Espermatozoides , Testosterona/análogos & derivados , Testosterona/sangueRESUMO
OBJECTIVE: To investigate the correlation of the single nucleotide polymorphism (SNP) rs4880 of the superoxide dismutase 2 (SOD2) gene with the risk of male infertility. METHODS: This caseîcontrol study included 519 male patients with idiopathic infertility (aged 19ï¼40 ï¼»28.93±4.93ï¼½ years) in the case group and 338 fertile men (aged 19ï¼40 ï¼»28.40±4.25ï¼½ years) in the control group. We collected the clinical data, genotyped the SNP rs4880 of the SOD2 gene by Sequenom Mass Array, and analyzed the association of different genotypes with male infertility using the logistic regression model. RESULTS: Statically significant differences were observed between the case and control groups in the level of follicleîstimulating hormone (FSH) (ï¼»4.72±2.51ï¼½ vs ï¼»15.65±17.24ï¼½ U/L, P< 0.01), the percentage of progressively mobile sperm (ï¼»9.12±13.5ï¼½ vs ï¼»41.95±9.03ï¼½%, P< 0.01), and sperm concentration (ï¼»12.95±24.38ï¼½ vs ï¼»72.88±45.60ï¼½ ×106/ml, P< 0.01), but not in other parameters. No correlation was found between male infertility and the heterozygous genotype TC (OR = 0.90, 95% CI: 0.65ï¼1.25, P = 0.516) or the homozygous genotype CC (OR=1.49, 95% CI: 0.38ï¼5.81, P = 0.566) as compared with the wild genotype TT, and similar results were obtained in the analysis of the subgroups. CONCLUSIONS: The SNP rs4880 of the SOD2 gene was not correlated with male infertility, which, however, is to be supported by further studies with larger samples from more areas.
Assuntos
Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Superóxido Dismutase/genética , Adulto , Estudos de Casos e Controles , Hormônio Foliculoestimulante/sangue , Predisposição Genética para Doença , Genótipo , Heterozigoto , Humanos , Modelos Logísticos , Masculino , Nucleotídeos/genética , Motilidade dos Espermatozoides , Adulto JovemRESUMO
Objective: To investigate the correlation of the single nucleotide polymorphismsï¼SNPsï¼ rs1799930 and rs1799931 of the N-acetyltransferase 2 geneï¼ NAT2ï¼ with the risk of male infertility in Nanjing area. Methods: We made a case-control study of 636 cases of male idiopathic infertility and 442 normal fertile men as controls. We genotyped the two SNPs by Sequenom Mass Array, analyzed the correlation of different genotypes with male infertility using the logistic regression model, and determined the association of the linkage effect of the two SNPs with male infertility by haplotype analysis. Results: Statistically significant differences were found between the case and control groups in sperm concentrationï¼[32. 32 ± 45. 49] vs [72. 77 ± 45. 21] × 106/ ml, P < 0. 01ï¼,the percentage of progressively motile spermï¼[15. 29 ± 5. 06] vs [42. 02 ± 9. 04]%,P < 0. 01ï¼,and the level of follicle-stimulating hormoneï¼[14. 69 ± 12. 37] vs [4. 72 ± 2. 51] U / L,P < 0. 01), but not in other parameters. No correlation was observed between the frequencies of the two SNPs or alleles in different models and male infertility. Haplotype analysis suggested a linkage effect within rs1799930 and rs1799931ï¼D' = 0. 998,r2= 0. 05ï¼ but no evident correlation between male infertility and genotype combination. Conclusion: The SNPs rs1799930 and rs1799931 of the NAT2 gene were not found to be correlated with the risk of idiopathic infertility in men.
Assuntos
Arilamina N-Acetiltransferase/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Alelos , Estudos de Casos e Controles , Genótipo , Haplótipos , Humanos , Modelos Logísticos , MasculinoRESUMO
OBJECTIVE: To determine the correlation of the CYP1A1 (rs4646422) gene polymorphisms with male infertility in the Chinese Han population. METHODS: Using the Mass ARRAY iPLEX GOLD technique, we conducted a case-control study on theCYPlA1 (rs4646422) gene polymorphisms in 636 infertile males aged 21-49 years (case group) and 442 normal healthy men aged 23-47 years (control group) of the Chinese Han population. We analyzed the genotypes and allele frequencies in the two groups ofsubjects with the SPSS 20.0 software. RESULTS: Compared with the wild homozygous genotype GG, the heterozygous genotype AG (OR = 1.06, 95% CI 0.81-1.38) and homozygous genotype AA (OR = 1.11, 95% CI 0.56-2.21) showed no correlation with male infertility, nor did the mutant allele A (OR = 1.06, 95% CI 0.85-1.32) in comparison with the wild allele G. CONCLUSION: The CYP1A1 (rs4646422) gene polymorphisms might not be correlated with male infertility in the Chinese Han population.
Assuntos
Citocromo P-450 CYP1A1/genética , Infertilidade Masculina/genética , Polimorfismo Genético , Adulto , Alelos , Estudos de Casos e Controles , China , Frequência do Gene , Genótipo , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Protamine (PRM) is one of the most abundant arginine-rich nucleoproteins in sperm and plays an important role in spermatogenesis. In the late stage of spermatogenesis, the replacement of PRM by histone prompts the closer combination between the nuclear matrix of sperm and nucleoprotein in order for high enrichment and condensation of nuclear chromatin in addition to preventing the sperm genome from mutation induced by internal and external factors. With the development of DNA sequencing techniques, researches on the association between PRM polymorphisms and male fertility are surfacing as a hot field. Many studies show that rs2301365 polymorphism is a risk factor for male infertility and increases the risk of male infertility by 27 - 66%, that rs737008 polymorphism of PRM1 and rs1646022 polymorphism of PRM2 are protective factors against Asian infertility, and that the ratio of PRM1 to PRM2 is intensively associated with male infertility. This review presents an update on the association between PRM gene polymorphisms and male infertility.
Assuntos
Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Protaminas/genética , Povo Asiático , Humanos , Masculino , Mutação , Fatores de Risco , Espermatogênese , EspermatozoidesRESUMO
BACKGROUND: 46,XX testicular disorder of sex development is a rare genetic syndrome, characterized by a complete or partial mismatch between genetic sex and phenotypic sex, which results in infertility because of the absence of the azoospermia factor region in the long arm of Y chromosome. CASE PRESENTATION: We report a case of a 14-year-old male with microorchidism and mild bilateral gynecomastia who referred to our hospital because of abnormal gender characteristics. The patient was treated for congenital scrotal type hypospadias at the age of 4 years. Semen analysis indicated azoospermia by centrifugation of ejaculate. Levels of follicle-stimulating hormone and luteinizing hormone were elevated, while that of testosterone was low and those of estradiol and prolactin were normal. The results of gonadal biopsy showed hyalinization of the seminiferous tubules, but there was no evidence of spermatogenic cells. Karyotype analysis of the patient confirmed 46,XX karyotype and fluorescent in situ hybridization analysis of the sex-determining region Y (SRY) gene was negative. Molecular analysis revealed that the SRY gene and the AZFa, AZFb and AZFc regions were absent. No mutation was detected in the coding region and exon/intron boundaries of the RSPO1, DAX1, SOX9, SOX3, SOX10, ROCK1, and DMRT genes, and no copy number variation in the whole genome sequence was found. CONCLUSION: This study adds a new case of SRY-negative 46,XX testicular disorder of sex development and further verifies the view that the absence of major regions from the Y chromosome leads to an incomplete masculine phenotype, abnormal hormone levels and infertility. To date, the mechanisms for induction of testicular tissue in 46,XX SRY-negative patients remain unknown, although other genetic or environmental factors play a significant role in the regulation of sex determination and differentiation.
Assuntos
Transtornos Testiculares 46, XX do Desenvolvimento Sexual/genética , Genes sry/genética , Transtornos Testiculares 46, XX do Desenvolvimento Sexual/patologia , Adolescente , Deleção de Genes , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Infertilidade Masculina/genética , Infertilidade Masculina/patologia , Inibinas/análise , Cariotipagem , Masculino , Fenótipo , Testículo/patologia , Vimentina/análiseRESUMO
OBJECTIVE: To study the relationship between metastasis or recurrence of hepatocellular carcinoma (HCC) and hepatitis B virus (HBV) DNA load or the presence of double mutation at 1762/1764 in the basic core promoter (BCP). METHODS: One-hundred-and-fifty-seven patients with HCC were included in the study. Events of tumor metastasis or recurrence were recorded during 120 weeks of clinical follow-up after treatment by surgery or transarterial chemoembolization (TACE). The 1-year follow-up included monthly alpha fetoprotein (AFP) measurement and abdominal ultrasonography (US), as well as helical computed tomographic (CT) scan performed every 3 months. Follow-up beyond 1-year (surveillance) included AFP measurement and abdominal US every 2 months and helical CT scan every 6 months. Suspected metastasis or recurrence was investigated by hepatic angiography and confirmed according to the combined imaging findings. Serum HBV DNA level was measured by real-time PCR. HBV genotypes were determined by PCR-restriction fragment length polymorphism analysis. RESULTS: Of the 157 HCC cases 110 experienced tumor metastasis or recurrence; the cumulative probability of post-treatment HCC metastasis or recurrence was 4 (2.55%) at week 12, 14 (8.92%) at week 24, 28 (17.83%) at week 48, 64 (40.76%) at week 72, 92 (58.60%) at week 96, and 110 (70.06%) at week 120. Multivariate analysis indicated that both the BCP 1762/1764 double mutations and HBV DNA levels were risk factors for HCC recurrence or metastasis. In particular, the incidence of HCC recurrence or metastasis increased with baseline serum HBV DNA levels in a dose-response manner, ranging from 8/19 (42.1%) for less than 3 log10 copies/ml HBV DNA to 35/61 (57.3%) for 3-5 log10 copies/ml and 67/77 (87.0%) for more than 5 log10 copies/ml. After adjusting for potential confounders, serum HBV DNA level remained independently associated with HCC metastasis or recurrence. HCC recurrence or metastasis occurred in 22/43 (51.2%) of patients without BCP 1762/1764 mutations and 88/114 (77.2%) of patients with BCP 1762/1764 mutations. The adjusted odds ratio for patients infected with BCP 1762/1764 double mutation HBV, compared with those infected with non-BCP 1762/1764 mutation HBV, was 5.264 (95% CI: 1.436-12.574, P less than 0.05). CONCLUSION: Infection with HBV carrying the BCP 1762/1764 double mutation and presence of high HBV DNA load are independent risk factors for developing HCC metastasis or recurrence after surgery or TACE.
Assuntos
Carcinoma Hepatocelular/patologia , Vírus da Hepatite B/genética , Neoplasias Hepáticas/patologia , Mutação , Adulto , Idoso , Carcinoma Hepatocelular/virologia , DNA Viral/sangue , Feminino , Genótipo , Antígenos do Núcleo do Vírus da Hepatite B/genética , Humanos , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Recidiva Local de Neoplasia , Regiões Promotoras Genéticas , Carga ViralRESUMO
AIM: To investigate the long-term prognosis in peptic ulcer patients continuing taking antithrombotics after ulcer bleeding, and to determine the risk factors that influence the prognosis. METHODS: All clinical data of peptic ulcer patients treated from January 1, 2009 to January 1, 2014 were retrospectively collected and analyzed. Patients were divided into either a continuing group to continue taking antithrombotic drugs after ulcer bleeding or a discontinuing group to discontinue antithrombotic drugs. The primary outcome of follow-up in peptic ulcer bleeding patients was recurrent bleeding, and secondary outcome was death or acute cardiovascular disease occurrence. The final date of follow-up was December 31, 2014. Basic demographic data, complications, and disease classifications were analyzed and compared by t- or χ2-test. The number of patients that achieved various outcomes was counted and analyzed statistically. A survival curve was drawn using the Kaplan-Meier method, and the difference was compared using the log-rank test. COX regression multivariate analysis was applied to analyze risk factors for the prognosis of peptic ulcer patients. RESULTS: A total of 167 patients were enrolled into this study. As for the baseline information, differences in age, smoking, alcohol abuse, and acute cardiovascular diseases were statistically significant between the continuing and discontinuing groups (70.8 ± 11.4 vs 62.4 ± 12.0, P < 0.001; 8 (8.2%) vs 15 (21.7%), P < 0.05; 65 (66.3%) vs 13 (18.8%), P < 0.001). At the end of the study, 18 patients had recurrent bleeding and three patients died or had acute cardiovascular disease in the continuing group, while four patients had recurrent bleeding and 15 patients died or had acute cardiovascular disease in the discontinuing group. The differences in these results were statistically significant (P = 0.022, P = 0.000). The Kaplan-Meier survival curve indicated that the incidence of recurrent bleeding was higher in patients in the continuing group, and the risk of death and developing acute cardiovascular disease was higher in patients in the discontinuing group (log-rank test, P = 0.000 for both). Furthermore, COX regression multivariate analysis revealed that the hazard ratio (HR) for recurrent bleeding was 2.986 (95%CI: 067-8.356, P = 0.015) in the continuing group, while HR for death or acute cardiovascular disease was 5.216 (95%CI: 1.035-26.278, P = 0.028). CONCLUSION: After the occurrence of peptic ulcer bleeding, continuing antithrombotics increases the risk of recurrent bleeding events, while discontinuing antithrombotics would increase the risk of death and developing cardiovascular disease. This suggests that clinicians should comprehensively consider the use of antithrombotics after peptic ulcer bleeding.
Assuntos
Fibrinolíticos/efeitos adversos , Úlcera Péptica Hemorrágica/tratamento farmacológico , Idoso , Doenças Cardiovasculares/complicações , Feminino , Seguimentos , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Úlcera Péptica/tratamento farmacológico , Úlcera Péptica Hemorrágica/mortalidade , Prognóstico , Modelos de Riscos Proporcionais , Recidiva , Estudos Retrospectivos , Fatores de Risco , Resultado do TratamentoRESUMO
Dyschromatosis symmetrica hereditaria (DSH) is a rare autosomal dominant pigmentary genodermatosis, which is characterized by a mixture of hyperpigmented and hypopigmented macules on the dorsal of the hands and feet, and on the face presented like freckle. Identification of RNA-specific adenosine deaminase 1 (ADAR1) gene results in DSH. This study was mainly to explore the pathogenic mutation of ADAR1 gene and provide genetics counselling and prenatal diagnostic testing for childbearing individuals.Mutational analysis of ADAR1 gene was performed by polymerase chain reaction (PCR) and electrophoretic separation of PCR products by 1.5% agarose gel electrophoresis. The coding exons and intron/exon flanking regions followed by bidirectional sequencing was performed on all participants. In this study, we found that a 28 year-old male patient harbouring a deleterious substitution of Leu1052Pro in the ADAR1 gene in a typical DSH family. His mother suffered from the DSH also owns the same mutation. This mutation, however, is not identified in the unaffected members in this family and those 200 normal controls. In summary, this new mutation Leu1052Pro reported here is pathogenic and detrimental for DSH. Our finding not only enriches mutation database and contributes to dissecting further the correlation between mutation position and phenotypical features of DSH, but also provides genetics counselling and prenatal diagnostic testing for childbearing couple.
Assuntos
Adenosina Desaminase/genética , Predisposição Genética para Doença , Transtornos da Pigmentação/congênito , Pigmentação/genética , Proteínas de Ligação a RNA/genética , Adulto , Análise Mutacional de DNA , Éxons/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Transtornos da Pigmentação/genética , Transtornos da Pigmentação/fisiopatologiaRESUMO
BACKGROUND: OCI-5, the rat homologene of human glypican 3 (GPC3), is confirmed upregulated in hepatocellular carcinoma (HCC). The present study was undertaken to detect gene expression change of OCI-5 during occurrence and progression of rat HCC. METHODS: Male Sprague-Dawley rats were given diethylnitrosamine (DENA) to induce HCC. Three DENA-induced rats and one control rat were sacrificed every week for 18 weeks during the development of HCC. Tissues specimens were snap-frozen in liquid nitrogen and total RNA was isolated. Sk-Hep1 cells were treated with DENA at different concentrations. The gene expression levels of OCI-5 and GPC3 were detected with the RT-PCR method. RESULTS: OCI-5 was not expressed in normal rat liver tissues. When HCC occurred and aggravated, OCI-5 expression was gradually elevated to a very high level. GPC3 was not expressed in the DENA-treated Sk-Hep1 cells. CONCLUSIONS: OCI-5 was not expressed in normal rat liver tissues but in rat HCC tissues. High-expression of OCI-5 in DENA-induced rat HCC model was the gene expression change of HCC not the DENA-induced gene expression. The expression level of OCI-5 was not only elevated in rat HCC but also gradually along the occurrence and progression of HCC, indicating that GPC3 might serve as a sensitive marker of early stage HCC.
Assuntos
Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Proteoglicanas de Heparan Sulfato/genética , Animais , Sequência de Bases , Carcinoma Hepatocelular/patologia , Modelos Animais de Doenças , Progressão da Doença , Glipicanas , Neoplasias Hepáticas Experimentais , Masculino , Dados de Sequência Molecular , RNA/análise , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Células Tumorais Cultivadas , Regulação para CimaRESUMO
OBJECTIVE: To evaluate the diagnostic value for pancreatic cancer of four serum tumor markers, carbohydrate antigen (CA) 199, CA242, CA50 and carcino-embryonic antigen (CEA), and fecal k-ras and p53 gene mutations. METHODS: From February 2002 to March 2004, 136 patients were consecutively diagnosed with pancreatic cancer in the three participating medical centers. The diagnosis was confirmed by pathology in 53 patients, of whom five were excluded because they did not have measurement of serum tumor marker. The remaining 48 patients comprised the case group in the study. Ninety-six patients with benign digestive diseases diagnosed during the same period were recruited as control subjects. They were matched by sex and age. In both groups, serum CA199, CA242, CA50 and CEA were measured by ELISA, and fecal k-ras and p53 gene mutations were measured by PCR-restriction fragment length polymorphism and PCR-single strand conformational polymorphism, respectively. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to compare their diagnostic value, as well as the sensitivity, specificity and likelihood ratio. Moreover, independent and sensitive tests from these non-invasive approaches were selected to form a parallel test that may have further improved sensitivity for diagnosis of pancreatic cancer. RESULTS: The AUC of serum CA199 and CA242 were 0.821 (95%CI 0.725-0.917) and 0.821 (95%CI 0.723-0.919), respectively. The optimal diagnostic value of serum CA199 for pancreatic cancer was 93 U/mL, with a sensitivity of 73.7% and specificity of 91.4%. The positive likelihood ratio of CA199 was 8.57, and the negative likelihood ratio was 0.29. The optimal diagnostic value of serum CA242 was 25 U/mL, with a sensitivity of 71.1% and specificity of 93.5%. The positive likelihood ratio of CA242 was 10.94, and the negative likelihood ratio was 0.31. The sensitivity of fecal k-ras gene mutation for diagnosis of pancreatic cancer was 77.4%, and the specificity was 81.2%. The positive and negative likelihood ratios of fecal k-ras gene mutation were 4.12 and 0.28, respectively. The sensitivity and specificity of fecal p53 gene mutation were 25.8% and 95.3%, respectively, and its positive and negative likelihood ratios were 5.49 and 0.78. The rate of fecal k-ras mutation was higher in patients with benign pancreatic diseases (57.14%) than that of controls with non-pancreatic disorders. The values of serum tumor markers and fecal k-ras and p53 gene mutation rates were not significantly different in subgroups according to site or stage of pancreatic cancer. The sensitivity and specificity of the parallel test of serum CA199 and fecal k-ras gene mutation were 94.06% and 74.22%, respectively, while the sensitivity and specificity of the parallel test of serum CA242 and fecal k-ras were 93.47% and 75.92%, respectively. CONCLUSIONS: Serum CA199 and CA242 are valuable diagnostic tools for pancreatic cancer. The diagnostic value is further improved when they are combined with fecal k-ras gene mutation measurement.
Assuntos
Biomarcadores Tumorais/sangue , Genes p53 , Genes ras , Neoplasias Pancreáticas/diagnóstico , Idoso , Antígenos Glicosídicos Associados a Tumores/sangue , Antígeno CA-19-9/sangue , Antígeno Carcinoembrionário/sangue , Fezes/química , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To determine the normal value of serum elastase 1 in Chinese adults and evaluate its diagnostic value for pancreatic cancer. METHODS: Serum elastase 1 and CA19-9 were measured in 132 samples, including 39 patients with pancreatic cancer, 48 with other gastrointestinal malignancy, 24 with gastrointestinal benign disease and 21 healthy adults as normal control. Multiple statistical methods including receiver operating characteristics curve and discriminant analysis were employed. RESULTS: The established normal range of serum elastase 1 in Chinese adults was found to be under 4.36 mg/L. Serum elastase 1 increased markedly in patients with pancreatic carcinoma of smaller size and/or located in the pancreatic head. The sensitivity, specificity and overall accuracy of elastase 1 for diagnosis of pancreatic cancer were 61.5%, 75.3% and 71.2%, respectively, as compared with 71.8%, 73.1% and 72.7% for CA19-9. Discriminant analysis can improve the sensitivity and overall accuracy of elastase 1 to 82.0% and 74.2%, respectively, with a slight decline in the specificity to 71.0%. CONCLUSIONS: The cutoff value of serum elastase 1 in normal Chinese adults is 4.36 mg/L. Serum elastase 1 is effective in the diagnosis of pancreatic cancer, especially for those of smaller size or in the pancreatic head. Use of appropriate statistical methods can help to make the diagnosis more accurate.
Assuntos
Biomarcadores Tumorais/sangue , Elastase Pancreática/sangue , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/enzimologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno CA-19-9/sangue , Estudos de Casos e Controles , Ensaios Enzimáticos Clínicos , Análise Discriminante , Feminino , Neoplasias Gastrointestinais/diagnóstico , Neoplasias Gastrointestinais/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To evaluate the diagnostic value of serum carcinoma markers CA19-9, CA242, CA50 and carcinoembryonic antigens (CEA), fecal K-ras and p53 gene mutation in pancreatic cancer. METHODS: We collected 136 new cases of pancreatic cancer and 240 patients with benign digestive diseases including 49 patients with benign pancreatic diseases diagnosed in Peking Union Medical College Hospital, Chinese Academy of Medical Sciences Tumor Hospital and Shenyang PLA General Hospital from February 2002 to March 2004. Blood samples were collected and serum carcinoma markers CA19-9, CA242, CA50 and CEA were measured. Fecal K-ras and p53 gene mutation were also measured. We decided the optimal cut-off points with receiver operating characteristic curves and calculated the areas under the curve (AUC). RESULTS: The AUC of serum CA19-9 and CA242 were 0.855 +/- 0.031 (95% CI 0.794 - 0.916) and 0.859 +/- 0.031 (95% CI 0.799 - 0.920) respectively. The optimal cut-off point for serum CA19-9 was 68 U/ml, with the sensitivity of 84.4% (98/116) and the specificity of 84.3% (145/172) for the diagnosis of pancreatic cancer. The optimal cut-off point for serum CA242 was 25 U/ml, with the sensitivity of 88.4% (84/95) and the specificity of 79.1% (144/182). The sensitivity of fecal K-ras mutation was 77.8%, and the specificity was 82.2%. The sensitivity and specificity of fecal p53 gene mutation were 27.8% and 95.2% respectively. The diagnostic scale of pancreatic cancer was calculated by four variables: serum CA19-9, CA242, fecal K-ras and p53, of which each variable deserved one point if measured positive. The optimal cut-off point for the scale was 2, and the AUC of the diagnostic scale was 0.946 +/- 0.017 (95% CI 0.912 - 0.980). CONCLUSIONS: Serum CA19-9 and CA242 are valuable diagnostic tools for pancreatic cancer. The diagnostic value will be further improved when they are combined with the measurement of fecal K-ras and p53 gene mutation.
Assuntos
Biomarcadores Tumorais/sangue , Genes p53 , Genes ras , Neoplasias Pancreáticas/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno Carcinoembrionário/sangue , Humanos , Pessoa de Meia-Idade , Mutação , Neoplasias Pancreáticas/genética , Curva ROC , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To develop a high risk scoring model and screening strategy to improve the diagnosis of symptomatic pancreatic cancer. METHODS: A hospital-based case-control study was undertaken among a cohort comprising 136 pancreatic cancer patients and 191 patients with benign gastrointestinal diseases who were hospitalized between Feb, 2002 and Mar, 2004. All patients were consulted with an epidemiological questionnaire. Risk factors and symptoms described in the questionnaire were compared between these two groups. Significant and borderline risk factors and symptoms were selected to undergo multivariate logistic regression. A high risk scoring model was constructed according to the weighted numerical scores of every variable. The diagnostic values of 4 tumor markers of pancreatic cancers (serum CA19-9, CA242, stool K-ras and p53 mutation) and 2 imaging tests (abdominal spiral CT and ultrasonography) were evaluated to provide evidence for establishing the diagnostic strategy. RESULTS: The average score was significantly higher for the pancreatic cancer patients than for the control patients [mean 49.6 (95% CI: 45.6-53.7) vs 21.6 (95% CI: 19.3-23.9); P < 0.01]. With a cutoff value of 27 points, the sensitivity and specificity of the scoring model was 87.0% and 68.1% respectively. CT had the highest sensitivity (94.7%) among the 4 tumor markers and 2 imaging tests. Combination of the two tumor markers (CA19-9 and stool K-ras) with CT or ultrasonography could improve the sensitivity to 100% with a specificity of 67.5%-73.0%. It was suggested that for high risk patients with a risk score more than 27, the combination test be recommended as the primary test, endoscopic ultrasonography (EUS) and/or endoscopic retrograde cholangipancreatography (ERCP) be considered for patients with inconclusive CT studies when risk score and tumor markers nevertheless suggest pancreatic cancer. CONCLUSION: The high risk scoring model provides a simple and feasible way to screen pancreatic patients in hospitals at all levels. Once high risk patients are identified, they can be transferred to higher level hospitals to receive further examinations. This screening strategy may help detect more resectable pancreatic cancers.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Pancreáticas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos Glicosídicos Associados a Tumores/sangue , Antígeno CA-19-9/sangue , Estudos de Casos e Controles , Colangiopancreatografia Retrógrada Endoscópica , Estudos de Coortes , Endossonografia , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Inquéritos e Questionários , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: To detecte whether loss of heterozygosity (LOH) at the MEN-1 locus as well as 22q occurs in sporadic insulinoma and if LOH can be used as a genetic marker to differentiate malignant and benign insulinomas. METHODS: MEN-1 gene and 22q allelotyping were performed by PCR with microsatallite markers in DNA from microdissected normal and tumor tissues from archived or frozen insulinomas (8 malignant and 32 benign, from 38 patients). The significance was calculated using t test and Cochran-Mantel-Haenszel Statistics, P < 0.05 was considered significant. RESULTS: Sixteen of the 40 insulinomas (40.0%) had MEN-1 LOH and 22q LOH was shown in 30 of the 40 tumors (75.0%). Eleven of the 30 tumors (37.0%) with 22q LOH had 22q12 LOH over a 3 cm region, whereas LOH in 14 tumors (47.0%) occurred at 22q13.3. Eight tumors with D22S 280 locus (22q12) LOH were shown without MEN-1 LOH while 14 of the 30 tumors without D22S280 LOH had MEN-1 LOH (0% vs 47%, P = 0.016). Six of the 14 tumors (43.0%) with 22q13.3 LOH were malignant, whereas 2 of 26 tumors without 22q13.3 LOH (8.0%) are malignant (P = 0.0088). CONCLUSION: MEN-1 gene LOH may contribute to a proportion of insulinomas, and 22q LOH occurs frequently in insulinomas. D22S 280 (22q12) LOH may independently contribute to the tumorogenesis of sporadic insulinoams, whereas detecting 22q13.3 LOH in insulinomas can be a potential genetic marker to distinguish malignancy from benign tumors.
Assuntos
Cromossomos Humanos Par 22/genética , Insulinoma/genética , Perda de Heterozigosidade , Neoplasias Pancreáticas/genética , Proteínas Proto-Oncogênicas/genética , Biomarcadores Tumorais/genética , Feminino , Marcadores Genéticos/genética , Humanos , Masculino , Neoplasia Endócrina Múltipla Tipo 1/genéticaRESUMO
OBJECTIVES: To investigate the differences in morphology, immunohistochemistry, DNA ploidy status, LOH and MSI of 11q13 and 1p between benign and malignant pheochromocytomas, and to find the marker or markers useful in distinction between benign and malignant pheochromocytoma or for predicting the malignant potential of this tumor. METHODS: Twenty-two cases of clinically documented benign and malignant pheochromocytomas from the files of Peking Union Medical College Hospital were analyzed. Aside from histological study, Ki-67, p53, CgA, S-100, PCNA and survivin immunohistochemistry studies were performed. DNA ploidy status was assessed by flow cytometry on cell suspensions prepared from formalin-fixed, paraffin-embedded sections. Twelve tumors (7 benign and 5 malignant) with paired normal tissues were microdissected. Tumor and normal tissue DNA were extracted. The obtained DNAs and 8 microsatellite markers related to 11q13 and 1q were subjected to PCR amplification for analysis of LOH and MSI. RESULTS: None of the tumors showed atypical mitosis, only 1 malignant tumor had a mitotic count > 1/10 HPF (2.3/10 HPF). Two malignant tumors exhibited confluent necrosis. Ki-67 index was low in benign tumors (average 0.73%), and high in malignant tumors (average 2.4%). The difference of Ki-67 index between benign and malignant tumors was statistically significant. DNA ploidy status did not correlate with malignancy. Although LOH and/or MSI of 11q13 and 1p were observed in several tumors, a statistically significant difference could not be reached due to the small number of tumors analyzed. CONCLUSION: Only Ki-67 index (> 3%) is an useful marker for distinguishing benign from malignant or for predicting the malignant potential of pheochromocytoma.