RESUMO
OBJECTIVE: To evaluate the value of a new platelet function test PFA P2Y (PFA-200) in monitoring clopidogrel treatment for cardiovascular disease in elderly patients. METHODS: Fifty-six elderly patients receiving clopidogrel therapy in the Department of Cardiology of General Hospital of PLA from March to August in 2016 and 85 healthy volunteers were recruited for analysis. All the subjects underwent PFA P2Y, LTA (light transmittance aggregometry) and TEG (Thromboelastograph) tests, and Spearman correlation coefficients were used to test the associations between test results. The agreement among the 3 platelet function test methods was assessed using Cohen's kappa coefficient. RESULTS: Correlation coefficient (r) was -0.701 (P<0.001) between PFA P2Y and LTA, and 0.475 (P<0.001) between PFA P2Y and TEG. The agreement was 75% between PFA P2Y and LTA and 67.9% between PFA P2Y and TEG. The κ value was 0.434 (P=0.001) between PFA P2Y and LTA and 0.242 (P=0.046) between PFA P2Y and TEG. With ADP-induced maximum platelet aggregation rate of LTA >50% as the laboratory clopidogrel resistance, the cut-off value of PFA P2Y was 119 s (AUC=0.733) with a sensitivity of 75.6% and a specificity of 73.3%. CONCLUSION: PFA P2Y has a moderate correlation and agreement with LTA, but has a poor correlation and agreement with TEG. PFA P2Y can be useful for assessing the effects of clopidogrel therapy and the association of the cut-off value (119 s) with the long-term clinical ischemic events needs be confirmed in further study.
Assuntos
Plaquetas , Doenças Cardiovasculares/tratamento farmacológico , Inibidores da Agregação Plaquetária/uso terapêutico , Testes de Função Plaquetária , Ticlopidina/análogos & derivados , Bioensaio , Testes de Coagulação Sanguínea , Clopidogrel , Humanos , Agregação Plaquetária , Sensibilidade e Especificidade , Ticlopidina/uso terapêuticoRESUMO
OBJECTIVE: In this study, we aimed to investigate the expressions of adhesion molecules on human bronchial epithelial cells and neutrophils in co-culture system, assess the effects of puerarin on suppressing these adhesion molecules expressions, and explore the roles of two crucial signal-transduction elements p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor kappa B (NF-κB) in modulating adhesion molecules expressions. METHODS: Neutrophils and BEAS-2B cells (one human bronchial epithelial cell line) were co-cultured, and adhesion molecules expressions on cell surface were detected using flow cytometry. The mRNA levels of adhesion molecules were assessed by real-time quantitative polymerase chain reaction (real-time qPCR). Phosphorylated p38 MAPK and inhibitor κB were analyzed by Western blot. RESULTS: In co-culture system, adhesion molecules expressions on BEAS-2B cells and neutrophils were enhanced significantly (P<0.05). Correspondingly, the mRNA levels of adhesion molecules were also increased greatly. Moreover, the pretreatment of peurarin obviously suppressed adhesion molecules expressions on cell surface. Furthermore, phosphorylated p38 MAPK and inhibitor κB in BEAS-2B cells and neutrophils were elevated in co-culture system, but decreased significantly after upon the treatment of peurarin (P<0.05). CONCLUSIONS: Coculture boosted the interactions between human bronchial epithelial cells and neutrophils mimicking airway inflflammation, whereas peurarin decreased the expression of adhesion molecules on cell surface by suppressing the activities of p38 MAPK and NF-κB pathways, and exhibiting its anti-inflflammation activity.
Assuntos
Brônquios/metabolismo , Moléculas de Adesão Celular/metabolismo , Isoflavonas/farmacologia , NF-kappa B/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Sequência de Bases , Brônquios/citologia , Brônquios/enzimologia , Bovinos , Linhagem Celular , Técnicas de Cocultura , Primers do DNA , Regulação para Baixo/efeitos dos fármacos , Células Epiteliais/enzimologia , Células Epiteliais/metabolismo , Neutrófilos/enzimologia , Neutrófilos/metabolismo , Fosforilação , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: Chronic kidney disease (CKD) is an established predictor of recurrent ischemic events in patients with coronary artery disease (CAD). This association has been partially ascribed to high post-treatment platelet reactivity (HPPR) according to platelet function testing. However, the influencing factors of HPPR are assay-dependent, and the relevant data of elderly patients with stable CAD are absent. PATIENTS AND METHODS: 310 elderly patients (>80years of age) with stable CAD taking prolonged maintenance clopidogrel (75mg/day) were studied. Maximal platelet aggregation rate (MPA%) with light transmittance aggregometry and Platelet Reactive Units (PRU) with VerifyNow (VN) P2Y12 system were obtained. Markers of platelet activation, including PAC-1 and CD62P, were also determined. RESULTS: Patients on different stages of CKD presented similar MPA% and expression of PAC-1 and CD62P. Although severe CKD patients were more likely to present HPPR identified by VNP2Y12 (odds ratio: 1.85, p=0.038), multiple logistic regression diminished this effect (adjusted odds ratio: 1.19, p=0.642), and revealed anemia as a possible predictor of HPPR (adjusted odds ratio: 5.92, p=0.001). However, in a parallel way, hemoglobin correlated with baseline PRU values as well as with post-treatment values (r=-0.624 and r=-0.463, respectively, p<0.001). Association between hemoglobin and PRU inhibition rate was not found. Moreover, hemoglobin exerted no influence on MPA% at all. CONCLUSION: CKD is not necessarily associated with reduced antiplatelet effects of clopidogrel in elderly patients with stable CAD taking prolonged maintenance clopidogrel, and the seemingly influence of CKD on HPPR assessed by VNP2Y12 assay may be due to the artifactual effect of hemoglobin on VNP2Y12.
Assuntos
Doença da Artéria Coronariana/diagnóstico , Doença da Artéria Coronariana/tratamento farmacológico , Inibidores da Agregação Plaquetária/uso terapêutico , Insuficiência Renal Crônica/diagnóstico , Ticlopidina/análogos & derivados , Idoso de 80 Anos ou mais , Plaquetas/efeitos dos fármacos , Clopidogrel , Doença da Artéria Coronariana/complicações , Creatinina/metabolismo , Feminino , Hemoglobinas/metabolismo , Humanos , Masculino , Razão de Chances , Selectina-P/metabolismo , Ativação Plaquetária , Agregação Plaquetária/efeitos dos fármacos , Receptores Purinérgicos P2Y12/metabolismo , Análise de Regressão , Insuficiência Renal Crônica/complicações , Fatores de Risco , Ticlopidina/uso terapêutico , Resultado do TratamentoRESUMO
OBJECTIVE: To study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection. METHODS: E. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast. RESULTS: A total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79. CONCLUSION: CTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.
Assuntos
Bacteriemia/microbiologia , Infecção Hospitalar/microbiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Escherichia coli/enzimologia , Escherichia coli/isolamento & purificação , Cirrose Hepática/terapia , beta-Lactamases/genética , Farmacorresistência Bacteriana , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Genótipo , Hospitalização/estatística & dados numéricos , Humanos , Testes de Sensibilidade Microbiana , beta-Lactamases/metabolismoRESUMO
OBJECTIVE: To investigate the correlation between pro coagulation factors and anti-coagulation factors synthesized by the liver, and the correlation between fibrin degradation products (FDP) and D-dimer (D-D) concentration and coagulation proteins synthesized by extra-hepatic tissues, in different liver diseases; to explore the relationship between coagulation and bleeding in hepatic diseases. METHODS: Chronic hepatitis B (CHB) patients, CHB-related liver cirrhosis patients, CHB-related liver failure patients and healthy (normal) controls were selected for study and provided blood samples for analysis. The activity of coagulation factors (F) II, V, VII, VIII, IX, X, XI, and XII was detected using the one-stage clotting method. Coagulogram analysis, including activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT), was conducted by the solidification method. Antithrombin III (AT-III) and protein C (PC) activities were measured by chromogenic substrate assay. FDP concentration was detected using immunoturbidimetry. Tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), von Willebrand factor (vWF), and tissue factor (TF) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: With the exception of FVIII, coagulation factors and anticoagulant proteins synthesized by the liver were decreased and the coagulogram was extended for all patients. Likewise, the FDP and D-D concentrations were increased in blood. CHB patients, however, presented with increased levels of FVIII, TFPI, TM, vWF, and TF. Pairwise comparison indicated statistical differences existed among CHB, CHB-related liver cirrhosis, and liver failure patients: TFPI: 239.3+/-206.4, 315.0+/-258.6, and 319.5+/-298.1 -- higher than normal control: 104.0+/-87.1, F = 5.453, P less than 0.05; vWF: 70.3+/-29.5, 105.5+/-58.0, and 179.3+/-61.7 -- higher than normal control: 21.9+/-7.2, F = 20.104, P less than 0.05; TF: 85.9+/-85.7, 234.2+/-202.9, and 344.7+/-214.6 -- higher than normal control: 12.8+/-8.1, F = 8.619, P less than 0.05; FVIII: 157.2+/-53.4, 206.9+/-86.9, and 335.7+/-117.7 -- higher than normal control: 105.5+/-46.2, F = 13.418, P less than 0.05. CONCLUSION: In parallel to the progression of liver diseases, pro coagulation and anti-coagulation elements synthesized by the liver were reduced. In contrast, fibrinolysis activity was enhanced, which is expected to lead to an imbalance between blood clotting and anti-clotting factors. This may be an important cause for the bleeding that occurs in end-stage liver disease. Expressions of TFPI, TM, vWF, and TF significantly change in the early stage of liver diseases, as compared to normal (healthy) levels, and may represent a sensitive indicator of vascular injury.
Assuntos
Fatores de Coagulação Sanguínea/metabolismo , Insuficiência Hepática/fisiopatologia , Hepatite B Crônica/fisiopatologia , Adulto , Idoso , Antitrombina III/metabolismo , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , Insuficiência Hepática/sangue , Hepatite B Crônica/sangue , Humanos , Hidrocarbonetos Clorados/metabolismo , Lipoproteínas/metabolismo , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Fator de von Willebrand/metabolismoRESUMO
OBJECTIVE: To detect the effect of extracellular Ca2+ concentrations on test results of coagulation-related parameters. METHODS: Blood samples of outpatient medical volunteers were collected and then different doses of calcium chloride added. The rate of platelet aggregation (n = 42), prothrombin time (PT), thrombin time (TT) and activated partial thromboplastin time (APTT) (n = 21) and parameters of thromboelastography (n = 30) were detected according to the standard protocols by plasma turbidimetry, coagulation and recalcification respectively. RESULTS: When the plasma Ca2+ concentration was in the range of 0.1 - 33.7 mmol/L, the rate of platelet aggregation gradually increased with a increasing concentration of Ca2+. And the rates induced by adenosine diphosphate (ADP) and arachidonic acid (AA) were (51.8 +/- 9.6)% - (94.7 +/- 4.8)% and (64.4 +/- 12.2)% - (93.2 +/- 5.5)% respectively. When the Ca2+ concentration was 39.0 mmol/L, the rate decreased markedly [ADP (9.1 +/- 5.3)%, AA (11.1 +/- 4.5)%, both P < 0.01]. When the Ca2+ concentration was in the range of 0.1 - 33.7 mmol/L, the values of PT gradually increased with a increasing concentration of Ca2+. The values of TT changed in "V"-type and became minimum when the calcium concentration was 4.4 mmol/L. The values of APTT decreased with higher calcium concentrations and could not be determined when the concentration increased above 0.5 mmol/L. When the Ca2+ concentration was in the range of 0.4 - 27.3 mmol/L, the values of reaction time and coagulation time of thromboelastography changed in "V"-type and became nearly minimal at the Ca2+ concentration of about 2.1 mmol/L. The values of alpha angle and maximum amplitude changed in "V"-type and became maximal at the Ca2+ concentration of 2.1 mmol/L. CONCLUSIONS: The effect of Ca2+ concentration on the testing results of coagulation-related parameters is significant. A high calcium ( > or = 39 mmol/L) can inhibit the platelet aggregation, coagulation factor activity and blood coagulation. The Ca2+ concentration of 2.1 mmol/L seems to be the optimal concentration for thromboelastography by recalcification method.
Assuntos
Cálcio/sangue , Agregação Plaquetária , Tempo de Protrombina , Tromboelastografia , Idoso , Coagulação Sanguínea , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tempo de Tromboplastina Parcial , Tempo de TrombinaRESUMO
OBJECTIVE: To identify asthenozoospermia-associated proteins in seminal plasma by the shotgun proteomic strategy. METHODS: Six seminal plasma samples were collected by Percoll respectively from healthy fertile and asthenozoospermia volunteers, balanced, mixed, and then the mixture was separated by SDS-PAGE. The proteins in the gel were enzymolyzed, extracted and identified by the shotgun proteomic strategy. The identified proteins with the unique peptide count > or =2 or the unique peptide count=1 but the total count > or =4 were compared between the two groups. RESULTS: A total of 172 differential proteins were identified, of which, 89 were exclusively from the asthenozoospermia and 83 exclusively from the healthy fertile men. According to the molecular function, these differential proteins were mainly the types of signal transduction and catalytic activity. CONCLUSION: Functionally, 10 of the proteins are particularly important, which include annexin VI isoform 2, isoform 1 of interleukin-6 receptor subunit beta precursor, Mr 400,000 protein, cytosolic dynein heavy chain, alpha-actinin-4, receptor-type tyrosine-protein phosphatase eta precursor, vitamin D-binding protein precursor, protein S100-A11, protein S100-A9 and ANXA4.
Assuntos
Astenozoospermia/fisiopatologia , Proteômica , Sêmen/química , Proteínas de Plasma Seminal/isolamento & purificação , Adulto , Eletroforese em Gel de Poliacrilamida , Humanos , Masculino , Proteína de Ligação a Vitamina D/isolamento & purificaçãoRESUMO
OBJECTIVE: To identify differential proteins in the seminal plasma of healthy fertile men and non-obstructive azoospermia patients by the shotgun proteomic strategy. METHODS: Six seminal plasma samples from 3 healthy fertile and 3 non-obstructive azoospermia volunteers were collected by Percoll isolation, balanced-mixed, and followed by separation of the mixture by SDS-PAGE. The proteins were subjected to in-gel enzymolysis and isolation of peptide fragments, and then identified by the shotgun proteomic strategy. Then comparative analyses were made between the two groups on the identified proteins with the unique peptide count > or = 2 and = 1 but with the peptide count > or = 4. RESULTS: A total of 213 differential proteins were identified, 133 in the non-obstructive azoospermia patients and 80 in the healthy fertile men. According to the molecular function, these differential proteins mainly fell into the types of signal transduction, cytoskeleton and catalytic activity, especially oxidoreductase activity in the latter type. Eighteen of the differential proteins were found to be of particular significance, including dynein heavy chain, fatty acid synthase, and tubulin alpha-6 chain. CONCLUSION: The differential proteins identified in this study were many in number and various in function, which not only demonstrated the value of the shotgun proteomic strategy in protein identification, but also suggested the complicated pathogenesis and varied types of non-obstructive azoospermia. The samples must be selected strictly based on their gene and histological types. Non-obstructive azoospermia was shown to be related with the M phase of the mitotic cell cycle at the protein level, but its specific mechanism remains unknown.
Assuntos
Azoospermia/metabolismo , Proteoma/análise , Sêmen/química , Azoospermia/fisiopatologia , Estudos de Casos e Controles , Humanos , Masculino , Proteômica/métodos , Motilidade dos EspermatozoidesRESUMO
OBJECTIVE: To identify proteins in the seminal plasma of healthy fertile men. METHODS: Three seminal plasma samples were collected from healthy fertile volunteers by Percoll isolation, and then the balanced mixture of the seminal plasma was separated by SDS-PAGE. The proteins in the gel band underwent enzymoloysis, and was extracted and identified by shotgun proteomic strategy. RESULTS: A total of 331 proteins were identified, with the molecular weight (MW) ranging from 8 000 to 572 068 and the isoelectric point (pI) from 4.36 to 11.05. Based on the molecular function and biological process of the proteins, 51 (15.4%) were classified as transport proteins, 11 (3.32%) as cell movement proteins, 63 (19.03%) as signal transduction proteins, 147 (44.4%) as proteases, 38 (11.5%) as enzyme regulator proteins, 21 (6.3%) as programmed cell death proteins, 12 (3.62%) as structural proteins and 59 (17.8%) as proteins with unknown molecular function. CONCLUSION: Shotgun proteomic strategy is a good method for protein identification. Annexin A, Annexin-associated proteins and the Ras-related protein Rab were the major members of the signal transducer proteins identified. Ca2+ and G protein signal pathways may play a most important role in the extracellular signal transduction into cells, but the interactions between these proteins remain unknown. The great quantity of enzymes and enzyme regulator proteins identified in the seminal plasma may be closely related with the maintenance of sperm motility and metabolism.
Assuntos
Fertilidade , Proteômica/métodos , Sêmen/química , Proteínas de Plasma Seminal/isolamento & purificação , Adulto , Humanos , Masculino , Motilidade dos EspermatozoidesRESUMO
OBJECTIVE: To analyse the variability of proteins in the seminal plasma of severe oligospermic and healthy fertile men. METHODS: Spermatic fluid samples were collected from 11 healthy fertile men and 6 severe oligospermic male volunteers and tested by SELDI-TOF-MS with the CM10 protein chip to get the protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc. RESULTS: The mean peak heights of 2 lower-abundance proteins expressed in the seminal plasma of the severe oligospermic men were statistically different from the healthy fertile males (P<0.05). Fifteen different proteins existed between the nonobstructive azoospermic and the severe oligospermic group, 7 of which, with m/z of 7,196.058, 7,547.610, 5,780.493, 7,059.844, 7,409.589, 5,379.173 and 10,778.810, also between the non-obstructive azoospermic and the healthy fertile males (P<0.05). Except the latter two, the contents of the other 5 proteins were decreased in the non-obstructive azoospermic men (P<0.05). CONCLUSION: The finger prints of the seminal plasma proteins of the severe oligospermic group were similar to those of the healthy fertile males, both significantly different from the non-obstructive azoospermic men. It is suggested that pathogenesis mechanisms differ exist between non-obstructive azoospermia and severe oligospermia but are not the simple accumulation of genetic factors.
Assuntos
Oligospermia/metabolismo , Sêmen/metabolismo , Proteínas de Plasma Seminal/análise , Adulto , Humanos , Masculino , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
OBJECTIVE: To improve the diagnostic ability of routine laboratory items in liver diseases associated with viral hepatitis through constructing assessment models consisting of these items. METHODS: (1) Assessment of routine items and formulation of models. Data of 447 patients seen between May 1997 and August 2003 were collected as the training set and serum specimens of 213 patients taken between June 2004 and March 2005 were examined and used as the validation set. Eleven items (TP, ALB, TBIL, DBIL, ALT, AST, ALP, GGT, TBA, LDH, CHE) were examined with an automated biochemical analyzer. Logistic regression was applied to construct the model for discriminating between chronic hepatitis and liver cirrhosis. The diagnostic value of items and models was assessed by the area under the receiver-operating characteristic (ROC) curve. RESULTS: The model to discrimination between chronic hepatitis and liver cirrhosis consists of five items (CHE, DBIL, ALB, ALT, GLO). The AUCs of model were 0.87 in the training set and 0.83 in validation set, respectively. CONCLUSION: (1) The model consisting of CHE, DBIL, ALB, ALT, GLO improves the diagnostic value of routine laboratory items in discriminating chronic hepatitis from liver cirrhosis.
Assuntos
Hepatite B Crônica/diagnóstico , Cirrose Hepática/diagnóstico , Adolescente , Adulto , Algoritmos , Diagnóstico Diferencial , Feminino , Humanos , Fígado/patologia , Fígado/fisiopatologia , Fígado/virologia , Cirrose Hepática/virologia , Testes de Função Hepática , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
OBJECTIVE: To investigate the difference in coagulation screening in heparin anticoagulant therapy between dry-chemistry point of care tests (POCT) and automatic analyzer. METHODS: Blood samples were collected from 44 patients with myocardial infarction or intracranial hemorrhage, 34 males aged 78 +/- 10 and 10 females aged 73 +/- 8, to undergo examination of prothrombin time (PT), international normalized ratio (INR), activated partial thromboplastin time (APTT), and fibrinogen (Fbg) by dry-chemistry methods with coagulation analyzers of the models CG02 and Hemochron Jr II. The levels of APTT of 30 patients were measured by the bedside with analyzers model Hemochron Jr II, after their anticoagulation samples underwent measurement of the plasma heparin activity with ACL Advance automatic coagulation analyzer; the APTT was determined with STA-R. RESULTS: The correlation coefficients of PT, INR, APTT, and Fbg between the CG02 and STA-R results were 0.945, 0.966, 0.662, and 0.977 respectively (all P < 0.05). However, t test showed that the levels of PT and INR between these 2 groups were not significant, and the differences in the levels of APTT and Fbg were significant (both P < 0.05). ACL Advance analyzer showed that the heparin activities of 30 samples were 172 approximately 1465 U/L, and the APTT values measured by Hemochron Jr II and STA-R were 87 s +/- 32 s and 90 s +/- 46 s (P < 0.01). CONCLUSION: Dry-chemistry POCT is not suitable to report the results of the same patient simultaneously with the automatic analyzer.
Assuntos
Anticoagulantes/uso terapêutico , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/tratamento farmacológico , Heparina/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , Testes de Coagulação Sanguínea/normas , Feminino , Humanos , Masculino , Sistemas Automatizados de Assistência Junto ao Leito/normas , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To analyse protein alterations in the seminal plasma of non-obstructive azoospermia patients. METHODS: Semen samples were collected from 11 healthy fertile and 6 azoospermia male volunteers respectively and tested by SELDI-TOF-MS with CM10 protein chip to get protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc. RESULTS: The mean peak heights of 28 proteins expressed in the seminal plasma of the azoospermia patients were statistically different from those of the healthy fertile males (P < 0.05 ), of which 24 were of lower contents than in the normal controls, 4 with remarkably significant difference, M/Z 7 196.058, 7 630.573, 7 547.610 and 7 709.833 (P < 0.01). CONCLUSION: The seminal plasma proteins of the azoospermia patients were significantly different from those of the healthy fertile males, with decreased contents of most of the different proteins, which might be significantly correlated with the development of azoospermia.
Assuntos
Azoospermia/metabolismo , Proteínas/análise , Proteômica/métodos , Sêmen/química , Adulto , Humanos , Masculino , Sêmen/citologia , Espectrometria de Massas por Ionização por Electrospray , Contagem de Espermatozoides , Motilidade dos EspermatozoidesRESUMO
OBJECTIVE: To analyse clinical feasibility of two electrophoresis procedures of seminal plasma proteins, agarose gel electrophoresis and SDS-agarose gel electrophoresis. METHODS: Sixty-nine semen samples were examined and classified into three groups: the asthenozoospermia (n = 22), the asthenoteratozoospermia (n = 19), and the relative normal group (n = 28) with normal routine and special test results, according to WHO routine and special test criterion. Then, the seminal plasma protenis were separated by two different electrophoresis, with SDS-agarose and agarose support medium, the buffer pH 7.0 and 9.2 respectively. The agarose gel electrophoresis was done under various sample loading time, motion power and staining modules. The completed gels were scanned and compared the each other statistically. RESULTS: Seminal plasma proteins can be separated into 4 strips by SDS-agarose gel electrophoresis with acid crystal violet, and the strips were diffusion and with dark background. However, 6 clear strips named A, B, C, D, E, and F can be obtained by agarose gel electrophoresis with 6 min. After samples were loaded and stained by amidoblack, there showed appropriate spaces among strips, and it was very easy to scan the drying gel by a densitometer. Using agarose gel electrophoresis, the statistical difference in strip C and E was shown between the asthenozoospermia and the relative normal group, and between the asthenozoospermia and the asthenoteratozoospermia, however, not between the relative normal and the asthenoteratozoospermia group. Moreover, the samples in the relative normal group with normal routine and special test results were in different electrophoresis maps. CONCLUSION: Agarose gel electrophoresis of seminal plasma proteins with buffer pH 9.2, 6 min. sample loading and amidoblack stain was a simple, fast and fit technique for clinic.
Assuntos
Eletroforese em Gel de Ágar/métodos , Eletroforese em Gel de Poliacrilamida/métodos , Proteínas/análise , Sêmen/química , Adulto , Humanos , Masculino , Coloração e RotulagemRESUMO
OBJECTIVE: To investigate the diagnostic value of coagulation factors in assessing the severity degree of liver cirrhosis caused by hepatitis B. METHODS: Fifty-eight patients with liver cirrhosis and twenty healthy persons as control were enrolled. Prothrombin time activity percentage (PTA), activated partial thromboplastin time, coagulation activity of factor II, V, VII, VIII, IX and X were detected with clotting assay. Antithrombin-III (AT-III) was detected with colorimetric assay. The biochemical markers were also detected. RESULTS: The differences of PTA, factor II, VII and AT-III among Child-Pugh A, B, C in patients with liver cirrhosis were statistically significant (P < 0.01). Through receiver operating characteristic curve analysis, when 64% and 50% were used as cut-off values for PTA and factor VII in diagnosing Child-Pugh B, the area under the curve (AUC) was 0.689 and 0.610, the sensitivity was 76.9% and 61.5%, the specificity was 62.2% and 55.6%; when 54% and 39% were used as cut-off values for PTA and factor VII in diagnosing Child-Pugh C, the AUC was 0.924 and 0.942, the sensitivity was 80.0% and 86.7%, the specificity was 88.4% and 90.7%. Stepwise linear regression was done between Child-Pugh grade and coagulation factors. PTA, cholinesterase (Che), total bilirubin (TBil), albumin (Alb), factor VII were included in regression equation, Y = 15.008 - 0.018 x PTA - 0.288 x Che + 0.264 x TBil - 0.988 x Alb - 0.034 x VII, R(2) = 0.871. Patients whose Y was less than 8 were classified as grade "a", between 8 - 10 as grade "b", more than 10 as grade "c", the diagnostic accuracy was 84.5%. CONCLUSION: Coagulation factor VII may serve as a helpful marker in diagnosing the severity degree of liver cirrhosis.
Assuntos
Fatores de Coagulação Sanguínea/análise , Hepatite B Crônica/complicações , Cirrose Hepática/sangue , Cirrose Hepática/diagnóstico , Adulto , Idoso , Antitrombina III/análise , Feminino , Humanos , Cirrose Hepática/etiologia , Masculino , Pessoa de Meia-Idade , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Sensibilidade e Especificidade , Índice de Gravidade de DoençaRESUMO
OBJECTIVE: To determine which expression mode of prothrombin time (PT) might achieve PT standardization in patients with advanced liver diseases. METHODS: PT was measured with six thromboplastins with different ISI values in 16 severe chronic hepatitis patients, 50 decompensated liver cirrhosis patients and 30 patients on oral anticoagulation therapy. The results were expressed in PT (second), PTA (%), PTR and INR. RESULTS: In chronic hepatitis patients, the means of the six group's PTAs ranged from 24% to 34%, while their upper limits ranged from 47% to 61%. The means of the INRs ranged from 2.55 to 5.13, while their upper limits ranged from 4.65 to 12.77. Through one-way ANOVA of repeated measures, PPTA (0.489) was > PINR (0.120). In patients with liver cirrhosis, the means of the PTA in six groups ranged from 50% to 59%, while their upper limits ranged from 82% to 90%. The means of the INR ranged from 1.40 to 1.80, while their upper limits ranged from 1.97 to 3.69. Through one-way ANOVA of repeated measures, PPTA (0.102) was > PINR (0.01). In patients on oral coagulation therapy, the means of PTA ranged from 26% to 37%, while their upper limits ranged from 39% to 49%. The means of INR ranged from 2.35 to 2.66, while their upper limits ranged from 3.16 to 4.26. Through one-way ANOVA of repeated measures, PPTA (0.01) was less than PINR (0.102). The correlation between the results detected by Neoplastine and by other reagents were analyzed. They correlated well with each other when PTA was used as the expression mode of PT in patients with advanced liver disease. But in patients on oral anticoagulation therapy, when only the INR was used as the expression mode of PT, the correlation was well with each other. CONCLUSION: The use of INR provides inadequate standardization. Only when the PT is expressed in PTA, then it may provide a standardization mode in patients with advanced liver diseases.