A case of inherited afibrinogenemia caused by an IVS7-12A>G splice mutation of FGG gene / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese pedigree affected with inherited afibrinogenemia.@*METHODS@#For the proband and his family members, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), Fibrin(ogen) degradation products (FDPs), D-dimer (D-D), plasminogen activity (PLG:A) and the TT mixed experiment with protamine sulfate were determined with a STAGO-R automatic coagulation analyzer. The activity and antigen of fibrinogen (Fg) in plasma were measured with the Clauss method and immunonephelometry method, respectively. All exons and flanking regions of the fibrinogen genes (FGA, FGB and FGG) were amplified by PCR and directly sequenced. Human Splicing Finder software was used to predict and score the change of splicing site caused by the mutation.@*RESULTS@#The proband showed normal FDPs and D-D but significantly prolonged TT, PT and APTT. The activity and antigen of fibrinogen in plasma were significantly decreased (G (g.4147A>G) mutations of the FGG gene, for which his parents and young sister were heterozygous. As predicted by Human Splicing Finder and Mutation Taster software, the variant may generate a new splicing site which can extend the sequence of exon 7 by 11 bp, with alteration of the coding sequence. PROVEAN suggested the variant to be deleterious.@*CONCLUSION@#The afibrinogenemia of the proband may be attributed to the FGG IVS7-12A>G variant, which was unreported previously.

Clinical and genetic analysis of a pedigree affected with type I hereditary antithrombin deficiency due to a g.2736dupT variant of the AT gene / 中华医学遗传学杂志

OBJECTIVE@#To analyze the phenotype and genotype of a patient affected with inherited antithrombin deficiency.@*METHODS@#All exons and exon-intron boundaries of the AT genes were subjected to PCR amplification and Sanger sequencing. The influence of variants on the disease was predicted using bioinformatic software (MutationTaster).@*RESULTS@#The results of all coagulation tests were normal, though the antithrombin activity and antigen content of the proband and his father have decreased significantly (34%, 48% and 12.97 mg/dL, 15.60 mg/dL, respectively). His mother was normal. Genetic analysis revealed that the proband and his father both carried a heterozygous g.2736dupT variant of the AT gene. Bioinformatic analysis suggested that the variant may be pathogenic.@*CONCLUSION@#The proband and his father both had type I hereditary antithrombin deficiency caused by a g.2736dupT variant of the AT gene. The variant was unreported previously.

Analysis of a pedigree affected with congenital dysfibrinogenemia due to a novel Gly31Glu mutation of FGA gene / 中华医学遗传学杂志

Objective@#To analyze the phenotype and genotype of a pedigree affected with congenital dysfibrinogenemia.@*Methods@#Liver and kidney functions of the proband and her relatives were determined. Coagulation tests including prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time(TT), fibrin(ogen) degradation products (FDPs), D-dimer(D-D) and the calibration experiment of protamine sulfate of against plasma TT were detected in the proband and her predigree members. The activity and antigen of fibrinogen (Fg) in plasma were measured by Clauss method and immunonephelometry method, respectively. All of the exons and exons-intron boundaries of the three fibrinogen genes (FGA, FGB and FGG) were subjected to PCR amplification and Sanger sequencing. Potential influence of the suspected mutations were analyzed with bioinformatics software including PolyPhen-2, SIFT and Mutation Taster.@*Results@#The proband had normal PT, APTT, FDPs, D-D and prolonged TT (31.8 s). The activity of fibrinogen (Fg) in plasma was significantly decreased but the antigen was normal. Genetic analysis revealed a heterozygous c. 92G>A (p.Gly31Glu) mutation in exon 2 of the FGA gene. Family studies revealed that the mother carried the same mutation. Bioinformatic analysis suggested that the mutation may affect the function of Fg Protein.@*Conclusion@#The dysfibrinogenemia was probably caused by the novel Gly31Glu mutation of the FGA gene.

Analysis of a pedigree affected with congenital dysfibrinogenemia due to a novel Gly31Glu mutation of FGA gene / 中华医学遗传学杂志

OBJECTIVE@#To analyze the phenotype and genotype of a pedigree affected with congenital dysfibrinogenemia.@*METHODS@#Liver and kidney functions of the proband and her relatives were determined. Coagulation tests including prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time(TT), fibrin(ogen) degradation products (FDPs), D-dimer(D-D) and the calibration experiment of protamine sulfate of against plasma TT were detected in the proband and her predigree members. The activity and antigen of fibrinogen (Fg) in plasma were measured by Clauss method and immunonephelometry method, respectively. All of the exons and exons-intron boundaries of the three fibrinogen genes (FGA, FGB and FGG) were subjected to PCR amplification and Sanger sequencing. Potential influence of the suspected mutations were analyzed with bioinformatics software including PolyPhen-2, SIFT and Mutation Taster.@*RESULTS@#The proband had normal PT, APTT, FDPs, D-D and prolonged TT (31.8 s). The activity of fibrinogen (Fg) in plasma was significantly decreased but the antigen was normal. Genetic analysis revealed a heterozygous c.92G>A (p.Gly31Glu) mutation in exon 2 of the FGA gene. Family studies revealed that the mother carried the same mutation. Bioinformatic analysis suggested that the mutation may affect the function of Fg Protein.@*CONCLUSION@#The dysfibrinogenemia was probably caused by the novel Gly31Glu mutation of the FGA gene.

Phenotypic and genetic analysis of two pedigrees affected with hereditary coagulation FXII deficiency / 中华医学遗传学杂志

OBJECTIVE@#To carry out phenotypic and genotypic analysis for two Chinese pedigrees affected with coagulation factor XII (F XII) deficiency.@*METHODS@#Plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), and blood coagulation factor VIII, IX, XI, XII activity (FVIII:C, FIX:C, FXI:C, FXII:C) were determined with one stage clotting assay on a STAGO coagulation analyzer. FXII antigen was determined with an enzyme linked immunosorbent assay (ELISA). The 14 exons and their flanking sequences of the F12 gene were subjected to PCR amplification and Sanger sequencing. The conservation and structure of mutant protein were analyzed with MegAlign software and PYMOL software.@*RESULTS@#The APTT of the probands was significantly prolonged, while their FXII:C and FXII:Ag were significantly reduced. Genetic analysis of the proband has revealed three novel mutations in the F12 gene, including g.5972G>A splice site mutation in intron 5, g.8810_8814delGTCTA in exon 14, and g.6259G>A (p.Pro182Leu) in exon 7. In addition, a previously known mutation IVS13-1G>A has been found.@*CONCLUSION@#Four mutations have been identified in the two Chinese pedigrees, among which three were novel. Above mutations probably played a role in the defect of FXII in the two pedigrees.

Genotypic and phenotypic analysis of a case with inherited coagulation factor X deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with FX deficiency.</p><p><b>METHODS</b>Prothrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen, FII activity(FII:C), FVII activity(FVII:C), FIX activity (FIX:C), FX activity(FX:C) were determined with a one-stage clotting assay. The FX antigen(FX:Ag) was detected with an enzyme linked immunosorbent assay(ELISA). The 8 exons, introns and 5' and 3' untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing. Suspected mutation was confirmed by reverse sequencing. Polymorphisms were excluded by direct sequencing of 100 healthy individuals.</p><p><b>RESULTS</b>The PT and APTT of the proband have prolonged to 16.1 s and 49.0 s, respectively. Her FX:C and FX:Ag were reduced by 27% and 56%, and her mother's PT, APTT, FX:C and FX:Ag were 14.8 s, 37.4 s, 44%, 34%, respectively. Her grandmother's PT, APTT, FX:C and FX:Ag were 15.8 s, 42.2 s, 31%, 45%, respectively. The results of her father and other family members were all within the normal range. Genetic analysis has revealed a heterozygous G to A mutation in the proband at position 28076 in exon 8 of the F10 gene, which resulted in a p.Gly363Ser substitution. The same mutation was also found in her mother and grandmother. No mutation of the F10 gene was found in her father. Gly363Ser may result in changes in the secondary structure of the FX protein and reduction of its activity.</p><p><b>CONCLUSION</b>The g.28076G to A(p.Gly363Ser) mutation of the F10 gene probably underlies the FX deficiency in this pedigree. The mutation was discovered for the first time in Chinese patients.</p>

Hyperhomocysteinemia Promotes Renal Function Impairment in Patients with Chronic Stable Coronary Artery Disease / 中国医科大学学报

Objective To investigate the possible mechanism of hyperhomocysteinemia (HHcy) in renal dysfunction in patients with chronic stable coronary artery disease (SCAD).Methods We consequentially enrolled patients diagnosed as having SCAD and,according to homocysteine (Hcy) levels,divided them into the HHcy group (Hcy≥15 μmol/L,n =53) and control group (Hcy＜15 μmol/L,n =47).We further tested the relationship among plasma Hcy level,renal function,and serum levels of interleukin (IL)-6 and adiponectin (APN).Results IL-6 was significantly higher in the HHcy group than in the control group (P ＜ 0.05).However,APN level was obviously lower in the HHcy group than in the control group (P ＜ 0.05).Correlation analysis revealed that Hcy level positively correlated with creatinine (r =0.379,P ＜ 0.001),uric acid (r =0.238,P =0.019),cystatin C (r =0513,P ＜ 0.001),and IL-6 levels (r =0.561,P ＜ 0.001) but negatively correlated with estimated glomerular filtration rate (eGFR;r =-0.288,P =0.023) and APN level (r =-0.428,P ＜ 0.001).IL-6 level showed positive correlations with creatinine (r =0.406,P =0.002),uric acid (r =0.359,P =0.007),and cystatin C levels (r =0.387,P =0.007) but a negative correlation with eGFR (r =-0.370,P =0.005).Meanwhile,APN level showed negative correlations with creatinine (r =-0.694,P ＜ 0.001),uric acid (r =-0.420,P ＜0.001),and cystatin C levels (r =-0.553,P ＜ 0.001),but a positive correlation with eGFR (r =0.251,P =0.034).Conclusion Plasma Hcy level may have a predictive value for renal dysfunction in patients with SCAD.Moreover,HHcy probably promoted renal dysfunction by inducing the imbalance of anti-inflammation and pro-inflammation in patients with SCAD.

Clinical and genetic analysis of 8 Chinese pedigrees with inherited dysfibrinogenemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze clinical manifestation and genetic mutations in 8 Chinese pedigrees featuring hereditary dysfibrinogenemia.</p><p><b>METHODS</b>Prothrombin time(PT), activated partial thromboplastin time(APTT), thrombin time(TT), calibration of plasma protamine sulfate against TT, fibrinogen (Fg) activity, coagulation factors II, V, VII, VIII, IX, X, XI and XII of all probands and their family members were detected with an automatic blood coagulation analyzer; D-dimer(D-D) and fibrin(ogen) degradation products(FDPs) were also dtected by automatic blood coagulation analyzer, Fg antigen were detected with an immunoturbidimetry method. Exons of fibrinogen genes FGA, FGB and FGG and flanking sequences were amplified by polymerase chain reaction(PCR) and sequenced.</p><p><b>RESULTS</b>All of the probands showed normal levels of FDPs, D-dimer(D-D) and activity of coagulation factor II,V,VII, VIII, IX,X,XI, XII. Plasma PT and APTT were normal or slightly prolonged. Prolonged TT was found in all of the probands, whilst TT was not significantly shortened by protamine sulfate. Fg antigen was within the normal range, but Fg activity was significantly decreased. The Fg antigen/activity ratio was greater than 2. One proband has carried a heterozygous variant of the FGA gene g.1233G>A(p.A α Arg35His). Four have carried a heterozygous mutation of the FGB gene g.9692A>G(p.Bβ Asn190Ser). The remaining 3 had heterozygous substitution of FGG gene g.10819G>A(p.γ Arg301His). In addition, 2 polymorphisms (p.A α Thr331Ala) and p.B β Arg478Lys) were identified in FGA and FGB genes.</p><p><b>CONCLUSION</b>p.A α Arg35His, p.B β Asn190Ser and p. γ Arg301His are responsible for the inherited dysfibrinogenemia in the 8 Chinese pedigrees. p.B β Asn190Ser is firstly reported in China. p.B β Asn190Ser and p. γ Arg301His may be mutation hot spot in the Chinese population.</p>

Association between CACNB2 gene polymorphisms and essential hypertension / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between single nucleotide polymorphisms (SNPs) of calcium channel β 2 subunit (CACNB2) gene and essential hypertension (EH) in ethnic Han Chinese in Wenzhou area, and to study the influence of rs7069292 alleles on gene expression with luciferase reporter technique.</p><p><b>METHODS</b>Sixty hundred and thirty seven Han Chinese with EH and 600 normal controls were enrolled. Genotypes of 6 SNP within CACNB2 gene including rs2228645, rs2357928, rs7069292, rs7099380, rs10764319 and rs11014166 were determined with matrix assisted laser desorption ionization/time of flight mass spectrometer (MALDI-TOF MS). A luciferase reporter gene plasmid containing the fragment flanking rs7069292 (-2831 bp to -2460 bp) in the 5' regulatory region of CACNB2 was constructed.</p><p><b>RESULTS</b>Compared with the control, CT and TT genotypes for the rs7069292 locus were significantly more common in EH group (5.20% vs. 2.17%, 2.59% vs. 1.08%, P< 0.05). CC genotype was not found. Promoter activity for allele C of the rs7069292 locus was significantly increased compared with allele T (P< 0.05). No significant difference was detected for other 5 SNPs in terms of genotypes and allele frequency.</p><p><b>CONCLUSION</b>The rs7069292 CT polymorphism of the CACNB2 gene is associated with EH in ethnic Han Chinese from Wenzhou area. A T>C mutation may affect the expression of CACNB2.</p>

Clinical features and treatment of type 1 diabetes mellitus in children / 中华全科医师杂志

Objective To review the clinical characteristics and treatment of type 1 diabetes mellitus (T1DM) in children. Methods The clinical data of 103 children with T1DM admitted to our hospital from Februry 2002 to August 2010 were retrospectively analyzed. Thirty one cases with diabetic ketoacidosis (KDA) were treated with continuous insulin pump (group A) or basal-bolus insulin therapy (group B). The differences in blood glucose control time, the rate of hypoglycemic episodes, glucose fluctuation, fasting blood glucose (FBG), 2 h postprandial blood glucose (2 hPBG), insulin dosage, the time of urine acetone bodies disappear and length of stay were compared in two groups. Results The age of 103 children with T1DM was from 38 d to 15. 33 y with an average of (8 ±3) y; most of them was 7 - 10 y (47, 45.6% ). Seventy eight children were first diagnosed accounting for 75.7%; boys accounted for 55.3% of total. Fifty one cases (49.5%) were diagnosed in winter and spring and 67 (65.2%) had infections, most of them were respiratory tract and gastrointestinal infections. Sixty two cases (60. 2% )presented as diabetic ketoacidosis at the first onset, including 4 cases (3.9%) with cerebral edema. Some patients were complicated by Hashimoto's thyroiditis, hyperthyroidism, SLE and other autoimmune diseases.Among 31 cases with ketoacidosis the FBG and PBG were decreased significantly after treatment, there were no significant differences between two groups (P ＞ 0. 05 ). Compared to group B the correction time of DKA and urine acetone bodies was shorter, and reached the targeted glucose levels more quickly, the frequency of blood fluctuation and the hypoglycemia was significantly lower, the length of stay was shorter, and the daily dose of insulin was lower in group A; the differences between two groups were statistically significant ( P ＜0. 05 or P ＜0. 01 ). Conclusions The clinical symptoms at first onset of T1 DM in children are not typical,and likely to be combined with DKA; infection may be one of the inducing factors for DKA. Continuous subcutaneous insulin infusion with pump can control the blood glucose more effectively and equably, and are convenient for use by children; so it is a better treatment option for type 1 diabetes mellitus in children.