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1.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-776756

RESUMO

OBJECTIVE@#To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis.@*METHODS@#The FⅦ antigen (FⅦ:Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ:C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank.@*RESULTS@#The propositus had prolonged PT (36.3 s), with FⅦ:C and FⅦ:Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ:C (86%-120%). The FⅦ:Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein.@*CONCLUSION@#The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.


Assuntos
Fator VII , Genética , Deficiência do Fator VII , Genética , Feminino , Testes Genéticos , Heterozigoto , Humanos , Masculino , Mutação , Linhagem
2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-776737

RESUMO

OBJECTIVE@#To analyze the phenotype and F5 gene variant in a pedigree affected with hereditary coagulation factor V (FV) deficiency.@*METHODS@#All of the exons, flanking sequences, and 5' and 3' untranslated regions of the F5 gene were subjected to PCR and direct sequencing. Suspected variant sites were confirmed by clone sequencing. Influence of the variants was predicted by using software including ClustalX and Mutation Taster.@*RESULTS@#The prothrombin time (PT) and activated partial thromboplastin time (APTT) of the proband were prolonged to 20.3 s and 59.2 s, respectively, while FV activity (FV:C) and FV antigen (FV:Ag) were reduced by 13% and 17%, respectively. The FV:C and FV:Ag of his father, sister and daughter were decreased to 35%, 37%, 29% and 42%, 46%, 35%, respectively. The proband was found to carry a heterozygous c.2851delT variant in exon 13 of the F5 gene, which caused a frameshift and resulted in a truncated protein (p.Ser923LeufsX8). In addition, a heterozygous c.1538G to A (p.Arg485Lys) variant was found in exon 10. The father, sister and daughter of the proband all carried the p.Ser923LeufsX8 variant, while his mother and son carried the heterozygous p.Arg485Lys polymorphism. His younger brother and wife were of the wild type. Bioinformatic analysis showed that p.Ser923 was highly conserved across various species. Mutation Taster scored 1.00 for the p.Ser923LeufsX8 variant, and the result has predicted a corresponding disease.@*CONCLUSION@#A heterozygous deletional mutation c.2851delT in exon 13 of the F5 gene and a heterozygous c.1538G to A polymorphism harbored by the proband may be associated with the decreased FV level in this pedigree.


Assuntos
Fator V , Genética , Deficiência do Fator V , Genética , Feminino , Deleção de Genes , Testes Genéticos , Heterozigoto , Humanos , Masculino , Mutação , Linhagem , Fenótipo
3.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-800863

RESUMO

Objective@#To analyze the phenotype and F5 gene variant in a pedigree affected with hereditary coagulation factor Ⅴ (FⅤ) deficiency.@*Methods@#All of the exons, flanking sequences, and 5′ and 3′ untranslated regions of the F5 gene were subjected to PCR and direct sequencing. Suspected variant sites were confirmed by clone sequencing. Influence of the variants was predicted by using software including ClustalX and Mutation Taster.@*Results@#The prothrombin time (PT) and activated partial thromboplastin time (APTT) of the proband were prolonged to 20.3 s and 59.2 s, respectively, while FⅤ activity (FⅤ∶C) and FⅤ antigen (FⅤ∶Ag) were reduced by 13% and 17%, respectively. The FⅤ∶C and FⅤ∶Ag of his father, sister and daughter were decreased to 35%, 37%, 29% and 42%, 46%, 35%, respectively. The proband was found to carry a heterozygous c. 2851delT variant in exon 13 of the F5 gene, which caused a frameshift and resulted in a truncated protein (p.Ser923LeufsX8). In addition, a heterozygous c. 1538G>A (p.Arg485Lys) variant was found in exon 10. The father, sister and daughter of the proband all carried the p. Ser923LeufsX8 variant, while his mother and son carried the heterozygous p. Arg485Lys polymorphism. His younger brother and wife were of the wild type. Bioinformatic analysis showed that p. Ser923 was highly conserved across various species. Mutation Taster scored 1.00 for the p. Ser923LeufsX8 variant, and the result has predicted a corresponding disease.@*Conclusion@#A heterozygous deletional mutation c. 2851delT in exon 13 of the F5 gene and a heterozygous c. 1538G>A polymorphism harbored by the proband may be associated with the decreased FⅤ level in this pedigree.

4.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-796468

RESUMO

Objective@#To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis.@*Methods@#The FⅦ antigen (FⅦ∶Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ∶C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank.@*Results@#The propositus had prolonged PT (36.3 s), with FⅦ∶C and FⅦ∶Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ∶C (86%-120%). The FⅦ∶Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein.@*Conclusion@#The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.

5.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-756409

RESUMO

Objective To observe confocal scanning laser ophthalmoscope (cSLO) based retinal imaging and color fundus camera in pigment epithelial detachment (PED) of polypoidal choroidal vasculopathy (PCV).Methods PED of 30 patients (32 eyes) were recruited from June 2016 to June 2017 in the Beijing Tongren Hospital who were detected in high-definition OCT (HD-OCT) and diagnosed as PCV by FFA and ICGA.There were 16 males (17 eyes) and 14 females (15 eyes);aged from 50-83 years,with the mean age of 66.59 years.The photographs of ocular fundus including color fundus camera,cSLO imaging,HD-OCT,FFA and ICGA were analyzed.Multimodal imaging results were regarded as gold standard.Sensitivity and specificity were calculated in serous and hemorrhagic PED diagnosis using color fundus camera and cSLO imaging.The positive number of PED was used to compare between two modes fundus imaging by using x2 test.Results Twenty serous PED eyes,3 hemorrhagic PED eyes and 9 serous/hemorrhagic PED eyes were determined using multimodal imaging.The sensitivity and specificity of color fundus camera were 45% and 100% in detecting serous PED and 100% and 91% in detecting hemorrhagic PED.The sensitivity and specificity of cSLO imaging were 83% and 100% in detecting serous PED and 50% and 86% in detecting hemorrhagic PED.The positive number of serous PED in cSLO imaging was significantly higher than color fundus camera (x2=7.752,P=0.011).The positive number of hemorrhagic PED in cSLO imaging shows no obvious difference compared with color fundus camera (x2=1.164,P=0.419).Conclusion The sensitivity and positive number of detecting serous PED with PCV in cSLO fundus imaging were higher than the color fundus camera technology.

6.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-772006

RESUMO

OBJECTIVE@#To identify potential mutations of F11 gene in a pedigree affected with hereditary coagulation factor XI (FXI) deficiency and explore its molecular pathogenesis.@*METHODS@#Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), coagulation factor VIII activity (FVIIIC), coagulation factor IX activity (FIXC), coagulation factor XI activity (FXIC), coagulation factor XII activity (FXIIC) and lupus anticoagulation (LA) of the proband and eight family members were determined. FXI antigen (FXIAg) was determined by enzyme-linked immunosorbent assay (ELISA). For the proband, potential mutations in the exons, flanking introns and 5'-, 3'-untranslated regions of the F11 gene were screened by direct DNA sequencing. The results were confirmed by reverse sequencing. Suspected mutations were detected in other family members. ClustalX-2.1-win and four online bioinformatic tools (PolyPhen-2, PROVEAN, SIFT, and Mutation Taster) were used to study the conservation and possible impact of the mutations. The structure of the mutational sites was processed with Swiss-PdbViewer.@*RESULTS@#The propositus had prolonged APTT (69.6 s), whose FXIC and FXIAg were reduced to 6.0% and 10.7%, respectively. Her mother, elder sister, one younger sister, little brother, daughter and son showed slightly prolonged APTT and moderate FXIC and FXIAg levels. Gene sequencing revealed that the propositus carried a heterozygous nonsense mutation c.738G>A (p.Trp228stop) in exon 7 and a heterozygous mutation c.1556G>C (p.Trp501Ser) in exon 13. Her mother, elder sister and daughter were heterozygous for the p.Trp228stop mutation, while one younger sister and little brother and son were heterozygous for p.Trp501Ser. Her husband and the youngest sister were of the wild type. Phylogenetic analysis suggested that Trp501 was highly conserved among all homologous species. The p.Trp501Ser was predicted to be "probably damaging","deleterious", "affect protein function" and "disease causing" corresponding to PolyPhen-2, PROVEAN, SIFT and Mutation Taster. Model analysis demonstrated that the non-polar Trp501 has two benzene rings, forming a hydrogen bond with Gln512 in the wild type. Once substituted by Ser501, the side chain may form another hydrogen bond with the benzene of His396. This may affect the normal space conformation and stability of FXI protein.@*CONCLUSION@#The compound heterozygous mutations of the F11 gene probably accounted for the low FXI concentration in this pedigree.


Assuntos
Fator XI , Genética , Deficiência do Fator XI , Genética , Feminino , Heterozigoto , Humanos , Masculino , Mutação , Linhagem , Filogenia
7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-745139

RESUMO

Objective To explore the application value of IL-8 monoclonal antibody microbubble combined with ultrsound targeted microbubble destruction ( UTMD) on alleviating myocardial ischemia reperfusion/injury ( MIRI) in rabbits .Methods Forty-two rabbits were randomly divided into closed chest group ( n =7) ,open chest control group ( n = 7) and ischemia-reperfusion ( I/R) group ( n = 28) .I/R group were randomly divided into 30 min reperfusion group( n =7) ,60 min reperfusion group( n =7) ,120 min reperfusion group ( n = 7 ) and 180 min reperfusion group ( n = 7 ) .All rabbits were examined by electrocardiogram , echocardiography and HE staining after MIRI . Targeted myocardial contrast echocardiography ( MCE) was performed and ELISA was used to detect IL-8 content in rabbit myocardium before and after UTMD . Results Electrocardiogram and wall motion returned to normal at 60 min after reperfusion .Targeted MCE showed that with the prolongation of reperfusion after I/R ,the video intensity of myocardium in reperfusion area increased gradually , reaching its peak at 120 min and 180 min after reperfusion .After UTMD ,the video intensity decreased ,and the change rate of video intensity in 30 min reperfusion group was higher than those in other reperfusion groups(all P<0 .05) .The content of IL-8 and its neutralization rate in the ELISA results were consistent with the video intensity and rate of change of targeted MCE .HE staining and scanning electron microscopy showed that myocardial injury was found in I/R group .With the prolongation of reperfusion time ,the degree of myocardial injury was gradually aggravated ,and the injury was alleviated after irradiation .Conclusions IL-8 monoclonal antibody combined with UTMD has the advantages of non-invasive and highly effective in alleviating MIRI .It provides a new way to treat MIRI .

8.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-807800

RESUMO

Objective@#To investigate the effect of hypertensive disorder complicating pregnancy (HDCP) on the mortality and early complications of premature infants.@*Methods@#The general clinical data of preterm infants with gestational age 24-36+ 6 weeks were collected from the cooperative units in the task group from January 1, 2013 to December 31, 2014.According to the severity of HDCP, the infants were divided into 4 groups: HDCP group, preeclampsia group, eclampsia group and non HDCP group, the mortality and major complications of preterm infants were compared, and the influencing factors were analyzed.@*Results@#The mortality rate of preterm in the HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ2=9.970, P=0.019). Eclampsia had a highest fatality rate (4.8%) in the early stage, compared with non HDCP group (2.2%), and the difference was statistically significant.Comparison of HDCP group (1.8%) and eclampsia group (3.2%) suggested that there was no statistically significant difference.The incidence of respiratory distress syndrome (RDS) in preterm in HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ2=13.241, P=0.004). Eclampsia group showed the highest incidence (35.4%), compared with non HDCP group (16.2%), the difference was statistically significant, but compared with HDCP group (19.9%), preeclampsia group (17.1%), there was no significant diffe-rence.The incidence of bronchopulmonary dysplasia (BPD) in preterm in HDCP group was significantly higher than that of non HDCP group (χ2=9.592, P=0.022), the highest incidence showed up in eclampsia group (9.7%), compared with non HDCP group (2.0%) and HDCP group (1.7%), the difference was statistically significant.But there was no statistically significant difference, compared with preeclampsia group.As the degree of HDCP aggravated, the incidence of BPD gradually rose.There was no significant impact on necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH) and sepsis of HDCP (χ2=7.054, 7.214, 0.358, 3.852; P=0.070, 0.065, 0.949, 0.278). Considering the overall outcome of the child, that was, whether the child died or survived, he had at least one complication, and HDCP had an effect on it (χ2=15.697, P=0.001), so the incidence increased while the degree of HDCP rose gradually.After adjusting gestational age, birth weight, sex, way of delivery, placental abruption and front placenta, prenatal hormonal, gestational diabetes, neonatal asphyxia and other factors, the results displayed that HDCP was the factor leading to the death of premature baby (OR=2.159, 95%CI: 1.093-4.266), and comparison between preeclampsia and eclampsia showed no statistical difference (P=0.714, 0.389); HDCP had no significant influence on RDS, BDP, ICH, NEC, ROP and sepsis.@*Conclusions@#HDCP leads to increased risk of premature death, but also leads to the increased incidence of RDS and BPD, but it had no obvious effect on NEC, ROP, IVH, sepsis and other complications.

9.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-699744

RESUMO

Objective This study was to evaluate the safety of 640 corneal donors by analysing the serological testing results.Methods We retrospectively analyzed the serological testing results from Changsha Aier Eye Bank and Chengdu Kangqiao Aier Eye Bank from January 2011 to December 2015,hepatitis B virus surface antigen (HBsAg),hepatitis C virus (HCV),treponema pallidum (TP) and human immunodeficiency virus (HIV) were detected by colloidal gold or enzyme-linked immunosorbent assay (ELISA).Results There were 83 out of 640 serum samples showed positive immuno-reaction assayed markers,the positive rate was 12.97%,including HBsAg(n=60,9.38%),HCV(n=3,0.47%),TP(n=11,1.72%) and HIV(n=2,0.31%).Moreover,3 corneal donors were both positive against HBsAg and HCV,2 donors positive against HCV and TP,1 donor positive against HBsAg and HIV,1 donor positive against HBsAg and TP.Conclusions There is a high proportion of positive results of blood-borne diseases in cornea donors,which is a potential threat to corneal receptors and eye bank workers.Therefore,it is very important to detect serological test strictly for corneal donors.

10.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-712148

RESUMO

Objective To analyze the phenotype and genotype of inherited dysfibrinogenemia pedigree associated with a novel heterozygous and deletion mutation in the FGG gene,and to investigate its molecular mechanism.Methods The clinical data were collected from the proband found at our hospital and her family members in April 2016.The activity plasma fibrinogen(Fg:C), activated partial thromboplastin time(APTT),prothrombin time(PT), thrombin time(TT)were detected by coagulation method and the antigen plasma fibrinogen(Fg:Ag), D-Dimer(D-D), fibrinogen degradation products (FDPs)were analyzed by immunoturbidimetry method.All of the exons and exon-intron boundaries of the genes of FGA, FGB and FGG with the fibrinogen(Fg)were amplified by PCR and followed by direct sequencing.And further verification were performed by cloning sequence and non-denatured polyacrylamide gel electrophoresis and silver staining.The conservatism of mutated gene locus were analyzed by ClustalX-2.1-win.The change of the protein spatial structure and the intermolecular forces with mutation were analyzed by Pymol.Results The Fg:C of the proband was significantly reduced(0.30 g/L)and the Fg:Ag of the proband was normal(2.00 g/L).Their Fg:C were both significantly reduced and the Fg:Ag were both normal(0.42 g/L,2.09 g/L & 0.47 g/L,2.42 g/L, respectively), these were found in her mother and grandma.Genetic analysis revealed a novel heterozygous and deletion mutation with c.944 _c.952 delCCTTTGATG in exon 8 of FGG gene in the proband,predicting a heterozygous 289_291delAla,Phe,Asp mutation.The same mutations were carried by her mother and grandma, but her father and grandpa were normal.Homology analysis indicated that the Ala 289,Phe290 and Asp291 were maintained highly conservative in homogenous species.Protein model analysis found that the original hydrogen bonds were disappeared when the deletion mutation happened with the Ala 289,Phe290 and Asp291.Conclusion The heterozygous and deletion mutation with 289_291delAla,Phe,Asp in the γchain of fibrinogen were identified that could cause the rearrangement of the Fg molecular space structure and the reduction of the structure stability,so the mutation probably underly the dysfibrinogenemia in this pedigree.(Chin J Lab Med,2018, 41:305-311)

11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-344126

RESUMO

OBJECTIVE To analyze the laboratory phenotype and FXII gene mutation in a consanguineous Chinese pedigree affected with factor XII (FXII) deficiency. METHODS Activated partial thromboplastin time (APTT), FXII activity (FXII:C) and FXII antigen (FXII:Ag) of the proband and her family members (10 individuals from 3 generations) were determined. Sanger sequencing was used to detect potential mutation within the 14 exons, their flanking regions and 5',3'-untranslated regions of the FXII gene. Suspected mutations were verified by backward sequencing. Conservation of the amino acids were analyzed with ClustalX-2.1-win. Four online bioinformatics software (PolyPhen-2, PROVEAN, SIFT and MutationTaster) were used to assess the impact of the mutations on the protein function. RESULTS The APTT of the proband and her elder brother have prolonged to 61.6 s and 68.6 s,and their FXII:C and FXII:Ag have decreased to 12%, 10% and 11%, 10%, respectively. The APTT of the paternal grandmother, maternal grandmother, father, mother, elder paternal aunt and elder maternal aunt were all normal, but their FXII:C and FXII:Ag have reduced to half of the normal value. Gene sequencing found that the proband and her elder brother have both carried a homozygous missense c.1078G>A (p.Gly341Arg) mutation in exon 10 of the FXII gene, for which the paternal grandmother, maternal grandmother, father, mother, elder paternal aunt and elder maternal aunt were heterozygous. Bioinformatic analysis suggested that the Gly341 is highly conserved, while p.Gly341Arg is a harmful mutation which may cause disease by affecting the function of FXII protein. CONCLUSION Homozygous p.Gly341Arg mutation, caused by consanguineous marriage, probably underlies the congenital FXII deficiency in this pedigree.

12.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-638259

RESUMO

Background Cystoid macular edema (CME) is caused by many fundus diseases.The noninvasive clinical diagnosis methods for CME are conventional color fundus photography up to now.However,these images can not display the CME range well.Confocal scanning laser ophthalmoscope (cSLO) based retinal imaging can provide clear picture with high contrast.However,whether cSLO imaging is feasible in the quantitative assessment of CME remains unclear.Objective This study was to image the boundary of CME and assess the quantification of CME image from cSLO imaging technology.Methods A series case-observational study was designed.This study protocal was approved by Ethic Committee of Beijing Tongren Hospital.cSLO based retinal imaging technology was carried out on consecutive 24 eyes of 24 patients with clinically diagnosed and OCT confirmed CME in Beijing Tongren Eye Center from August to December 2015 under the informed consent of each individual.The radial scan range was 45°× 45 ° and the line scan level was 49 at macula area.The pseudocolar image,green light reflective image (532 nm) and infrared reflective image (785 nm) were collected.The imaging was analyzed by EasyScan software (version 1.2.2).Fundus color photography and SD-OCT were carried out in each patient.The images were graded by specialists according to the SD-OCT cross sectional results.Results The primary causes of CME included epiretinal membrane (10 eyes),branch retinal vein occlusion (BRVO) (6 eyes),central retinal vein occlusion (CRVO) (4 eyes),diabetic retinopathy (DR) (3 eyes) and CRVO with BRVO (1 eye).A CME image was exhibited on the fundus color photogram with the obscure boundary;while the clear range of CME was displayed by the cSLO imaging.The mean score of CME from pseudocolar image,green light reflective image and infrared reflective image was 3.21±0.78,2.67±0.96 and 2.54±0.83,respectively,which was significantly higher than 1.33±0.82 from the fundus color photography (all at P<0.01).Conclusions In CME patients,the imaging quality from cSLO-based retinal imaging technology is better than that from traditional fundus color photography.Combined with SD-OCT sectional scan analysis,cSLO-based retinal imaging technology may offer a method to observe and record more fundus details for CME diagnosis.

13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-508303

RESUMO

Hydrogen spectrum nuclear magnetic resonance (1 H-NMR ) is a commonly used method for metabolomics and has been applied in the study of liver cirrhosis and liver cancer,however,study on serum metabolites in patients with primary biliary cholangitis (PBC)is rare.Aims:To investigate the capability of 1 H-NMR for screening serum metabolites of PBC.Methods:Twenty PBC patients,20 HBV-related cirrhosis patients and 20 healthy controls were detected by 1 H-NMR.The 1 H-NMR spectra data were processed by principal component analysis (PCA)and orthogonal partial least squares-discriminant analysis (OPLS-DA)so as to create the diagnostic model.Based on the correlation coefficient P (corr),VIP value and non-paired t test of OPLS-DA model,the differential metabolites between normal group and the two cirrhosis groups were screened.Results:OPLS-DA model could effectively distinguish healthy controls and PBC patients,model interpretation ability and prediction ability were 81.9% and 44.8%,respectively (P=0.0293), glutamine,folic acid,urocanic acid,4-ethylbenzoic acid were differential metabolites.OPLS-DA model could also effectively distinguish healthy controls and HBV-related cirrhosis patients, urocanic acid, 1-methylhistamine, 1-methyladenine,glucose,L-acetylcarnitine were differential metabolites.OPLS-DA model could not effectively distinguish PBC patients and HBV-related cirrhosis patients.Conclusions:Serum glutamine and folic acid may be the potential biomarkers of PBC,which may be closely related to the immune damage mechanism and prognosis of PBC;1 H-NMR combined with OPLS-DA diagnostic model are expected to become a new method for studying liver cirrhosis.

14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-345335

RESUMO

<p><b>OBJECTIVE</b>To explore the pathogenesis of protein C deficiency in two pedigrees through mutation detection and model analysis.</p><p><b>METHODS</b>Chromogenic substrate method and enzyme linked immunosorbent assay (ELISA) were used to determine the plasma protein C activity (PC: A) and protein C antigen (PC: Ag) in the two probands and their family members. All of the 9 exons and intron-exon boundaries of the PROC gene were amplified by PCR and analyzed with Sanger sequencing after purification. Corresponding mutate sites of the family members were also amplified and sequenced. The PolyPhen-2 software was used to analyze the perniciousness of the mutations and Clustal X was to analyze the conservatism. The protein model and amino acids interaction of the mutations were analyzed by Swiss-PdbViewer software.</p><p><b>RESULTS</b>The PC: A and PC: Ag of proband 1 was 30% and 35%, while PC:A of his father, mother and aunt were all slightly under the reference range. Two heterozygous missense mutations were found in exons 7 and 5 of the PROC gene, namely c.565 C>T (p.Arg147Trp) and c.383 G>A (p.Gly86Asp). His father and aunt were carriers for c.565 C>T, while his mother had carried c.383 G>A. The PC: A of proband 2 and his son were 50% and 64%, respectively. And they were both positive for p.Arg147Trp. Analysis of PolyPhen-2 indicated that p.Arg147Trp was benign, while p.Gly86Asp was damaging. Clustal X analysis indicated that the p.Arg147Trp was non-conservative, while the p.Gly86Asp was highly conservative. Modeling for the mutant proteins revealed that the simple aromatic ring of Trp147 in p.Arg147Trp destroyed the two hydrogen bonds between Arg147-Lys146 and Arg147-Lys151, and steric hindranted with Arg178. The side chain of Asp86 extended and generated steric clash with Gln90 with the occurrence of p.Gly86Asp. The change of hydrogen bonds and steric effects has altered the spatial configuration of amino acids, which led to unstable mutate proteins and interfered with the secretion.</p><p><b>CONCLUSION</b>Both probands had hereditary protein C deficiencies, for which their parents were all carriers. The heterozygous mutations p.Arg147Trp and p.Gly86Asp were the main cause for PC: A activity decrease. Among these, p.Gly86Asp was discovered for the first time.</p>


Assuntos
Sequência de Bases , Criança , Análise Mutacional de DNA , Métodos , Saúde da Família , Feminino , Heterozigoto , Humanos , Ligação de Hidrogênio , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Mutação , Linhagem , Fenótipo , Proteína C , Química , Genética , Metabolismo , Deficiência de Proteína C , Sangue , Genética , Domínios Proteicos
15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-497149

RESUMO

Objective To cpmpare the assessment of retinal and choroidal disease using confocal scanning laser ophthalmoscope (cSLO) imaging and color fundus camera.Methods Sixty-seven patients (90 eyes) with fundus diseases were included in this study.There were 35 males (51 eyes) and 32 female (39 eyes),mean age was 51.32 years.All subjects underwent fundus imaging using cSLO technology and traditional color fundus camera,positive numbers of every retinal pathological change were calculated and compared.Spectral domain-optical coherence tomography (SD-OCT) was also done to compare the accordance rate between two modes of fundus imaging (cSLO technology and traditional color fundus camera) and SD-OCT in choroidal changes.Results The positive numbers of retinal microaneurysm (x2 =4.157,P< 0.05) and epiretinal membrane (x2 =5.428,P < 0.05) using cSLO fundus imaging were significantly higher than traditional color fundus camera,while the positive numbers of cotton wool spots (x2 =0.523),retinal hemorrhage (x2 =0.117),hard exudates (x2 =0.325) and macular hole (x2 =0.070)were no significant different (P> 0.05).The SD-OCT accordance rate of choroidal pathological changes using cSLO technology was higher than traditional color fundus camera (x2 =9.143,P=0.007).Conclusion In retinal and choroidal diseases,the imaging quality of cSLO fundus imaging technology is better than the traditional color fundus camera technology.

16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-494288

RESUMO

Objective To detectthe phenotype and gene mutations underlying aninherited dysplasminogenemia pedigree and search the virulence gene.Methods The peripheral venous blood samples of the proband and his family members (fourteen subjects of three generations in total) were collected,and their prothrombin time(PT),activated partial thromboplastin time(APTF),thrombin time(TT),fibrinogen (FIB),fibrinogen degradation products (FDP),D-dimmer (D-D)weretested on a STAGO analyzer,the plasminogen activity (PLG:A) and plasminogen antigen (PLG:Ag) were analyzedby thechromogenic substrate assay and rocket immunoelectrophoresis,respectively.All 19 exons,5' and 3' untranslated regions of PLGwere amplified with PCR.Direct DNA sequencing was used to analyze the amplified products,which wereconfirmed by backward sequencing.Three bioinformatics online softwares (SIFT,PolyPhen-2 andMutationTaster) were used to forecast the possible impact of the mutations on the protein function.At last,themodel analysis of mutate site was taken on a Swiss-Pdb Viewer software.Results The PLG:Avalue of theproband and other 6 family members were decreased to the half,while the PLG:Ag was normal.The D-Dand FDP value of the proband,his grandma and father were slightly higher.DNA sequencing has revealedthat the proband and the other 6 members of this family had the same mutation of g.38829G > A in exon 15,leading to the missense mutationp.Ala601Thr.The results of bioinformatics softwares showed that themutation could affect the thePLGfunction.Protein model analysis indicated that the hydrophobic interaction force and hydrogen bond between the amino acids were changed,which might affect the stability of the PLG.In addition,all the members of this family take the heterozygous SNP of g.2501C > A in the 5 'UTR.Conclusions The p.Ala601Thr found in the inherited dysplasminogenemia pedigree in the exon 15 was responsible for the reduced PLG:A of the family,the dysplasminogenemia and this mutation were both reported for the first time in China.

17.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-491111

RESUMO

Objective To analyze the CRE and UREA result of university students,and establish the refer-ence interval for CRE,UREA adapted to this university.Methods Performing serum analysis of 2 926 freshmen for CRE,UREA with the automatic biochemical analyzer ( Olympus AU400 ) .Results Reference intervals were get among freshmen as CRE:74.00-109.00μmol/L (M),59.00-85.00 μmol/L (F),and it was different between male and female(Z =-27.27,P 24-45 years),1.94-5.11mmol/L ( female aged 25-45 years) .Conclusion The reference intervals disaggregated by sex and age are initially estab-lished.

18.
Chongqing Medicine ; (36): 915-917, 2016.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-490951

RESUMO

Objective To explore the clinical characteristics of patients with transjugular intrahepatic portosystemic shunt (TIPS) and literature review in patients with clinical features ,and provide clinical reference for carrying out the TIPS .Methods Totally 31 patients in our hospital from January 2009 to May 2014 who received TIPS treatment and strict follow‐up were retro‐spectively analyzed ,the preoperative basic situation ,laboratory index ,the incidence of postoperative bleeding again ,surgical compli‐cations ,the use of anticoagulant drugs and thrombosis ,dissolved ,etc .were statistical analyzed .Results In all patients with TIPS in the diagnosis of cirrhosis and portal hypertension ,hepatitis B ,hepatitis C cirrhosis and portal hypertension ,alcoholic liver cirrhosis and portal hypertension ,unknown cause of liver cirrhosis and portal hypertension ,Budd Chiari syndrome ,hepatitis B and hepatitis C cirrhosis and portal hypertension ,primary biliary cirrhosis and portal hypertension in proportion of 45 .16% ,16 .13% ,12 .90% , 12 .90% ,6 .45% ,3 .22% ,3 .22% respectively ;the incidence of postoperative bleeding again within six months was 9 .68% ;the Child‐Puhg score of preoperative and postoperative 1 week and 3 months ,6 months was (8 .35 ± 2 .52) ,(8 .32 ± 1 .76) ,(9 .29 ± 2 .55) ,(8 .10 ± 1 .85) respectively .Statistical results showed in postoperative 1 week and 3 months ,6 months ,there was no statisti‐cally significant difference compared with preoperative respectively (P>0 .05) ,postoperative 3 months liver function score of Child‐Puhg was higher than that of postoperative 1 week and 6 months (P<0 .05) operation;the rate of abdominal hemorrhage ,hepatic encephalopathy ,stent stenosis were 3 .22% ,22 .58% ,12 .90% ;the proportion of no postoperative taking anticoagulants ,taking as‐pirin ,clopidogrel ,and warfarin were 9 .68% ,38 .71% ,41 .94% ,9 .68% ,respectively ;the formation of portal vein thrombosis (inclu‐ding thrombosis increased) rate was 12 .90% ,thrombus dissolution rate was 100% .Conclusion In China ,liver cirrhosis and portal hypertension is the main source of TIPS and hepatitis B is a major cause of liver cirrhosis ;TIPS have no effect on liver function in Child‐Puhg score;hepatic encephalopathy ,stent restenosis is still the main postoperative complications of TIPS ;rules taking antico‐agulant drugs can dissolve thrombus of the portal vein and prevent thrombosis .

19.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-247720

RESUMO

<p><b>OBJECTIVE</b>To explore the phenotype, genotype and molecular mechanism for two pedigrees affected with hereditary antithrombin (AT) deficiency.</p><p><b>METHODS</b>Clinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, antithrombin activity (AT:A) and specific antigen (AT:Ag), protein C activity, as well as protein S activity. To detect potential mutations in the probands, all exons, exon-intron boundaries and the 3', 5' untranslated regions were amplified by PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and silver staining. The effect of mutations on the AT protein was analyzed with bioinformatics software.</p><p><b>RESULTS</b>The AT:Ag of pedigree 1 was normal, but its AT:A has reduced to 30%. A heterozygous c.235C>T mutation in exon 2 causing p.Arg47Cys, in addition with two single nucleotide polymorphisms (c.981G>A, c.1011G>A) in exon 5 were identified in the patient. His four children, except for the elder daughter, were heterozygous for the mutations. The plasma levels of AT:A and AT:Ag in proband 2 have decreased to 39% and 103 mg/L, respectively. A heterozygous deletion (g.5890-5892delCTT) leading to loss of p.Phe121 was also detected in his father. Bioinformatic analysis suggested that the missense mutation Arg47Cys can affect the functions of AT protein. Meanwhile, lacking of Phe121 will result in loss of hydrogen bonds with Ala124, Lys125 and the cation π interactions with Lys125, Arg47, which may jepordize the stability of the protein.</p><p><b>CONCLUSION</b>The proband 1 had type II AT deficiency, while proband 2 had type I AT deficiency. The p.Arg47Cys and g.5890-5892delCTT mutations of the AT gene are significantly correlated with the levels of AT in the two probands, respectively.</p>


Assuntos
Adulto , Idoso de 80 Anos ou mais , Antitrombina III , Genética , Metabolismo , Deficiência de Antitrombina III , Genética , Éxons , Feminino , Testes Genéticos , Genótipo , Humanos , Masculino , Mutação , Tempo de Tromboplastina Parcial , Linhagem , Fenótipo , Proteína C , Genética , Metabolismo , Proteína S , Genética , Metabolismo
20.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-247687

RESUMO

<p><b>OBJECTIVE</b>To identify potential mutation underlying coagulation factor X (FX) deficiency in a consanguineous Chinese pedigree.</p><p><b>METHODS</b>Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FX activity (FX:C) and other coagulant parameters were determined with a one-stage clotting assay. The FX antigen (FX:Ag) was determined with an ELISA assay. All coding exons and exon-intron boundaries of the F10 gene were amplified with PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and analyzed with CLC Genomics Workbench 7.5 software.</p><p><b>RESULTS</b>The PT and APTT in the proband were prolonged to 67.2 s and 102.9 s, respectively. Further study showed that her FX:C and FX:Ag were reduced by 1% and 8%, respectively. The PT of her father, mother, and little brother were slightly prolonged to 14.5 s, 14.4 s and 14.4 s, respectively. The FX:C and FX:Ag in her father, mother and little brother were all slightly reduced. Genetic analysis of the proband has revealed a homozygous G>A change at nucleotide 27881 in exon 8 of the F10 gene, which predicted a p.Val298Met substitution. The proband's father, mother, and little brother were all heterozygous for the p.Val298Met mutation. The proband has inherited the homozygous mutation from her parents by consanguineous marriage. Other family members were all normal. Bioinformatics analysis has indicated that this mutation may result in changes in the secondary structure of the FX protein.</p><p><b>CONCLUSION</b>A homozygous mutation g.27881G>A(p.Val298Met) of the F10 gene has been identified, which probably accounts for the low FX concentrations in this pedigree.</p>


Assuntos
Adulto , Sequência de Aminoácidos , Consanguinidade , Fator X , Genética , Deficiência do Fator X , Genética , Feminino , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Linhagem , Tempo de Protrombina
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