Identification of novel compound heterozygous variants in a pedigree affected with hereditary coagulation factor XI deficiency / 中华医学遗传学杂志

OBJECTIVE@#To analyze the phenotype and genetic basis for a pedigree affected with hereditary coagulation factor XI deficiency.@*METHODS@#Activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), FXI activity (FXI:C) and the antigen of FXI (FXI:Ag) were determined for the proband and members from his pedigree. Sanger sequencing was used to analyze all exons, exon-intronic boundaries, as well as the 5'- and 3'- untranslated regions of the F11 gene. Suspected variants were verified in her family members and confirmed by reverse sequencing. The impact of the variants on the protein function was predicted by using PolyPhen-2 and SIFT software. The protein structure and amino acid interaction were analyzed by using Swiss-PdbViewer.@*RESULTS@#The APTT, FXI:C and FXI:Ag of the proband and her sister were significantly reduced to 73.0 s, 10.0%, 15.0% and 87.1 s, 2.0% and 11.5%, respectively. APTT of some family members was slightly prolonged, and FXI:C and FXI:Ag also decreased to various extents. DNA sequencing revealed that the proband and her sister have carried compound heterozygous variants of c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) respectively in exons 7 and 9 of the F11 gene. Her father, sister and daughter were heterozygous for the c.738G>A (p.Trp228stop) variant, while her mother and nephew were heterozygous for the c.938G>T (p.Ser295Ile). Both PolyPhen-2 and SIFT predicted that the p.Ser295Ile variant is likely to be deleterious and can affect the protein function. Modeling analysis indicated that the p.Ser295Ile variant may lead to disruption of a hydrogen bond, resulting in alteration of protein structure and instability.@*CONCLUSION@#The compound heterozygous c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) variants of the F11 gene probably underlie the decreased FXI level in this pedigree.

Analysis of a pedigree affected with congenital hypofibrinogenemia due to heterozygous Ser313Ile mutation of fibrinogen γ chain gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic basis for a Chinese pedigree affected with congenital hypofibrinogenamia.</p><p><b>METHODS</b>Peripheral blood samples were collected from 9 members from the pedigree. Routine coagulation tests including activated partial thromboplastin time (APTT), thrombin time (TT), the prothrombin time (PT) were carried out. The activity of fibrinogen (Fg: C) was measured using Clauss method, and fibrinogen antigen (Fg: Ag) was measured with immunoturbidimetry. All exons and exon-intron boundaries of the fibrinogen Aα, Bβ and γ chain genes were amplified using PCR, which was followed by direct sequencing. Suspected mutation was confirmed by reverse sequencing. The mutant fibrinogen was analyzed with Swiss-PdbViewer.</p><p><b>RESULTS</b>The proband showed prolonged APTT, PT and TT. Her functional fibrinogen (Fg: C) and antigen fibrinogen (Fg: Ag) levels were reduced to 0.69 g/L and 0.72 g/L, respectively. Her mother and grandmother also had a low levels of fibrinogen, which were 0.99 g/L and 0.83 g/L for Fg: C, 1.02 g/L and 0.87 g/L for Fg: Ag, respectively. The results of other members from the pedigree were all within the normal range. Genetic analysis reveled a heterozygous G>T mutation at nucleotide 7590 in exon 8 of γ gene in the proband, which was predicted to be a novel Ser313Ile mutation. The mutation was also found in her mother and grandmother. Model analysis showed that the Ser313Ile mutation disturbed the hydrogen bonds between Ser313, Asn319 and Asp320. Moreover, the mutation also altered the mutual electrostatic force and affected the folding and instability of the mutant fibrinogen.</p><p><b>CONCLUSION</b>The heterozygous Ser313Ile mutation probably underlies the hypofibrinogenemia in this pedigree.</p>

Curative effect of sclerotherapy injection combined with somatostatin on esophageal variceal bleeding / 中国基层医药

Objective To explore the effect,safety and prognosis of sclerotherapy combined with somatostatin in the treatment of esophageal varicose hemorrhage.Methods From January 2014 to January 2017, a total of 100 patients with esophageal varicose hemorrhage in the Central Hospital of Yiwu were selected ,and they were divided into control group and observation group according to the digital table ,with 50cases in each group.The control group was treated with somatostatin alone , the observation group was treated with somatostatin combined with hardener injection.The clinical efficacy, safety and prognosis were compared between the two groups.Results Before treatment,the esophageal varices width had no statistically significant difference between the two groups ( t＝0.86, P＞0.86).After treatment,the esophageal varices width of the two groups were improved ( t＝2.64,4.82,all P＜0.05),the esophageal varices width of the observation group [(0.74 ±0.32)cm] was significantly better than (0.93 ± 0.32)cm of the control group (t＝2.31,P＜0.05).The blood transfusion volume of the observation group [(324.94 ± 37.52)mL] was significantly lower than (512.46 ±62.48)mL of the control group (t＝13.86,P＜0.05).The hemostatic time of the observation group [(15.24 ±2.36)h] was significantly shorter than (21.31 ±3.29)h of the control group (t＝4.65,P＜0.05).The hospitalization time of the observation group [(11.73 ±2.39) d] was significantly shorter than (20.75 ±3.46)d of the control group (t＝6.97,P＜6.97).The hemostasis of the observation group was 38 cases,and the hemostatic rate was 76%,which was significantly higher than 52% of the control group (χ2＝5.63,P＜0.05).The number of patients with postoperative fever ,poststernal pain,swallowing obstruction,sick and vomit,palpitation in the observation group was lower than those in the control group ,but there was no statistically significant difference (χ2＝0.95,1.03,0.86,1.25,0.92,all P＞0 0.05).The incidence rate of adverse reaction of the observation group was 22%,which was lower than 40%of the control group (χ2＝6.43,P＜0.05).The rebleeding rate of the observation group was 12%,which was significantly lower than 34%of the control group (χ2＝5.63,P＜0.05).Conclusion The injection of sclerosing agent in the treatment of esophageal varicose hemorrhage has good clinical efficacy, safety, less complications and better prognosis , which is worthy of application and widespread promotion.

Correlation between B lymphocyte activation and CD154 expression and the generation of lupus anticoagulants in patients with systemic lupus erythematosus / 中国医师进修杂志

Objective To explore the correlation between B lymphocyte activation and CD154 expression and lupus anticoagulants (LAC) in patients with systemic lupus erythematosus (SLE). Methods Sixty newly diagnosed SLE patients (SLE group) and 32 healthy controls (control group) were involved. The expressions of CD27 and CD154 in peripheral blood were determined by flow cytometry, and the positive expression rates were computed. The LAC was determined by activated partial thromboplastin time, and the LAC ratio > 1.20 was positive. Results The positive rate of CD27, expression intensity of CD27, positive rate of CD154 and expression intensity of CD154 in SLE group were significantly higher than those in control group: 0.047 ± 0.021 vs. 0.035 ± 0.014, 0.387 ± 0.185 vs. 0.214 ± 0.091, 0.191 ± 0.108 vs. 0.101 ± 0.081 and 0.049 ± 0.018 vs. 0.022 ± 0.012, and there were statistical difference (P<0.05 or <0.01). Among patients with SLE, the LAC positive was in 28 cases, and the LAC negative was in 32 cases. The positive rate of CD27, expression intensity of CD27, positive rate of CD154 and expression intensity of CD154 in SLE patients with LAC positive were significantly higher than those in SLE patients with LAC negative: 0.055 ± 0.023 vs. 0.037 ± 0.014, 0.444 ± 0.179 vs. 0.329 ± 0.123, 0.218 ± 0.101 vs. 0.158 ± 0.044 and 0.058 ± 0.035 vs. 0.020 ± 0.009, and there were statistical differences (P<0.05). Conclusions The B lymphocyte activation and CD154 abnormal expression correlates with generation of LAC in patients with SLE. It provides a basis for the further study on intervening the generation of LAC and reducing the risk of thromboembolism.

Analysis of phenotypes and genetic mutations in two pedigrees affected with hereditary protein C deficiency / 中华医学遗传学杂志

<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>

Phenotype and genotype analysis of in herited dysplasminogenemia caused by plasminogen Ala601Thr mutation / 中华检验医学杂志

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.

Val247Leu and Trp316Ser poly morphisms of β2-glycoprotein Ⅰ in systemic lupus erythematosus patients and their associations with antiphospholipid antibodies and thrombotic complications / 中华风湿病学杂志

Objective To study the Val247Leu and Trp316Ser polymorphisms of β2-glycoprotein Ⅰ(β2GPⅠ) in systemic lupus erythematosus (SLE) patients and their associations with antiphospholipid antibodies and thrombotic complications.Methods We used DNA sequencing to detect the polymorphisms of Val247Leu and Trp316Ser in 378 SLE patients and 240 normal controls.Anti-β2GP Ⅰ antibodies and anticardiolipin (ACA) were tested by enzyme linked immunosorbent assay (ELISA).Lupus-type anticoagulants(LAC) was performed by diluted Russell's Viper Venom Test.Then the patient group was further analyzed according to APLs (Anti-β2GP Ⅰ antibody,LAC and ACA),thrombosis and obstetrical complications using Logistic regression analysis to confirm whether there are associations between β2GPⅠpolymorphism and those factors.Results For Va1247Leu,the predominant genotype was LL in healthy controls which accounted for 57.08％,while it was VL in SLE patients which accounted for 59.5％ (x2=45.01,P=0.000).Frequency of VV genotype was significantly higher in patients with thrombosis,anti-β2GP Ⅰ,ACA and obstetrical complications (OR=6.79,3.75,2.12 and 3.85,respectively;P=0.000,0.001,0.044 and 0.017,respectively).Those patients with VL genotype tended to have positive anti-β2GPI,LAC,ACA,thrombosis and also obstetrical complications (OR=2.95,1.88,2.47,2.97 and 2.74,respectively;P=0.000,0.007,0.000,0.001 and 0.016,respectively) than those negative ones.The predominant genotype of Trp316Ser was TT,then TS.No correlations could be found between Trp316Ser polymorphism and APLs,neither relation to thrombosis complications.Conclusion The polymorphism of Val247Leu is significantly associated with the presence of APLs,thrombosis and obstetrical complications.Both VV and VL genotype are risk factors for the generation of APLs,occurrence of thrombosis and obstetrical complications.The VV genotype is a high risk factor for thrombosis.Trp316Ser polymorphism does not contribute to the APLs production and also have no correlations with thrombotic complication.

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

<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>

Fifty-two Cases Analysis of Cliff Suicide / 法医学杂志

Objective To analyze suicide scene, damage morphology and case investigation of mountain type scenic spot for giving the references to determine the nature of cliff suicide cases. Methods The suicide cases collected from 2002 to 2012 in scenic spot of Mount Huangshan. The age, gender, native place, case location, damage inspection and behavior were analyzed. Results In the 52 suicide cases, the suicide rate of male was higher than that of female. The numbers from other provinces were higher than that of local province. The age was mainly range from 19 to 50. The time of suicide cases mostly happened between 16:00 to 24:00. The major damage was compound injury with varying degrees of traumatic brain injury, organic injury of pleuroperitoneal cavity and surface bruise and scratch. Conclu-sion In order to determine the nature of cliff suicide cases, it needs to work synthetically in the investi-gation of crime scene and interview.

Analysis of molecular pathogenesis and clinical phenotypes in 10 probands with inherited fibrinogen deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the molecular pathogenesis and clinical phenotypes in 10 probands with inherited fibrinogen (Fg) deficiency.</p><p><b>METHODS</b>The diagnosis of hereditary Fg deficiency was validated by prothrombin time (PT), thrombin time (TT), Fg activity (Fg:C) and Fg antigen (Fg:Ag) in plasma. All of the exons and their flanking sequences of the Fg gene were analyzed by direct sequencing. Detected mutations were confirmed by reverse sequencing.</p><p><b>RESULTS</b>The ranges of Fg:C and Fg:Ag in the 10 probands were 0.52-0.91 g/L and 0.62-2.98 g/L, respectively. Five of the probands had type I disorders, and 5 had type II disorders. Seven point mutations were identified, among which 6 have located in the D region. γThr277Arg, γAsp316His, γTrp208Leu and Lys232Thr were novel mutations, and αArg19Ser was first reported in Chinese. Four probands had the same mutation site (γArg275). As to the clinical manifestation, probands with type I disorders were asymptomatic or with mild or medium symptoms, while those belonged to type II disorders had moderate or serious symptoms. Two probands have carried an Arg275Cys mutation but had different clinical manifestations.</p><p><b>CONCLUSION</b>Mutations of the Fg gene seem to aggregate to the D region of FGG in our region, and Arg275 is a common mutation. However, no correlation has been found between the mutation site and clinical manifestations.</p>

Expression of heme oxygenase-1 in lung tissue of paraquat poisoned mice and its significance / 中华危重病急救医学

ObjectiveTo investigate the expression of heme oxygenase-1 (HO-1) in lung tissue of mice with acute paraquat poisoning, and discuss its pathological mechanism.Methods Fifty-eight healthy male mice were randomly divided into control group (n = 8) and poisoned group (n = 50). The mice in poisoned group were lavaged with 20% paraquat (50 mg/kg), and those in control group with equal amount of normal saline. The mice were sacrificed on the day of experiment in control group, and those in poisoned group at 6 hours and 1, 3, 7, 14 days after poisoning. The lung tissue was harvested to observe the changes in pathology of lung with hematoxylin and eosin (HE) staining. The positive expression of HO-1 was determined with immunohistochemistry, and the protein expression of HO-1 was determined with Western Blot. The contents of superoxide dismutase (SOD) and malonaldehyde (MDA) were determined.Results The mice showed shortness of breath and signs of exhaustion 1 hour after poisoning, getting worse on 3-5 days, but returned to normal 14 days after poisoning. Under the light microscope, it showed that the control group had no significant pathological changes in lung tissue. One day after the ingestion, pulmonary alveolar structure disorder, obvious hemorrhage, edema and infiltration of inflammatory cells were found. At 3 days, the pathological changes in the lung tissue were more pronounced. They were less pronounced on 7 days, and inflammatory changes disappeared on 14th day, but alveolar structure disorder remained. Immunohistochemical test showed that HO-1 was seldom expressed in the lung tissue, and a little amount was expressed in the mucosal epithelial cells of the airway in control group. It was shown that inflammatory cell and endothelial were mainly distributed in the mucosal epithelial cells of airway 1 day after poisoning followed by a gradually decrease tendence, and came to normal level of control group 7 days after poisoning. It was shown by Western Blot that HO-1 (gray value) in lung tissue increased 6 hours after poisoning (2.438±0.467 vs. 0.475±0.167,P 0.05). The SOD activity (μU/L) in lung tissue was lowered 6 hours after poisoning, and it was significantly lower than that of control group (649.681±13.951 vs. 1 167.051±15.744,P 0.05).Conclusion HO-1 expression was increased significantly in lung tissue of mice with acute paraquat poisoning, which may be considered as an important protection mechanism against paraquat poisoning.

Genetic analysis of a pedigree with hereditary coagulation factor Ⅶ deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential mutations in a family affected with inherited factor Ⅶ (FⅦ) deficiency.</p><p><b>METHODS</b>Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FⅦ activity (FⅦ:C) and other coagulant parameters of the proband and 15 family members were measured. Potential mutations were screened in the pedigree by polymerase chain reaction and direct DNA sequencing.</p><p><b>RESULTS</b>The PT of the proband and his younger brother was significantly prolonged to 39.0 s and 30.1 s, respectively. FⅦ:C of the proband and his younger brother was obviously reduced to 2% and 3%, respectively. FⅦ:C of his grandmother, maternal grandmother, aunt, father, mother, maternal uncle and maternal aunt was all below the normal range (80%-108%), which measured 68%, 54%, 71%, 73%, 62%, 72% and 59%, respectively. The other coagulant parameters were in the normal range. Two heterozygous mutations, g.11349G>A and g.11482T>G, both reside in exon 8 of the F7 gene, have resulted in p.Arg304Gln and p.His348Gln substitutions, were identified in the proband. The same mutations were also found in the proband's younger brother. Four maternal members in this family (grandmother, mother, maternal uncle and maternal aunt of the proband) were heterozygous for the p.Arg304Gln mutation, while three paternal members (grandmother, aunt and father of the proband) were heterozygous for the p.His348Gln mutation.</p><p><b>CONCLUSION</b>The proband had inherited two independent mutations of the F7 gene including g.11349G>A and g.11482T>G from his mother and father, respectively. The compound heterozygous mutation probably explains the low FⅦ concentrations in this pedigree.</p>

Congenital hypofibrinogenemia associated with a novel mutation in FGG gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the genetic mutation underlying congenital hypofibrinogenamia in a Chinese pedigree.</p><p><b>METHODS</b>Standard coagulation tests including the prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), plasminogen activity (PLG:A), D-Dimer (DD) and fibrin degradation products (FDP) were tested with fresh plasma using a STA-R analyzer. The activity of fibrinogen (Fg:C) and fibrinogen antigen (Fg:Ag) were measured respectively with the Clauss method and immunoturbidimetry. All exons and exon-intron boundaries of the fibrinogen Aα-, Bβ-, and γ-chain genes (FGA, FGB and FGG) were amplified by PCR followed by direct sequencing. Suspected mutation was confirmed by reverse sequencing and analyzed with a Swiss-PdbViewer.</p><p><b>RESULTS</b>The PT level in the proband was normal, while the APTT and TT were slightly prolonged. The functional and antigen fibrinogen levels were both significantly reduced (0.91 g/L and 0.95 g/L, respectively). Similar abnormalities were also found in her father, elder sister, son and niece. The coagulant parameters of her mother were all within the normal range. Genetic analysis has reveled a heterozygous A>C change at nucleotide 5864 in exon 7 of γ gene in the proband, predicting a novel Lys232Thr mutation. The proband's father, elder sister, son and niece were all carriers of the same mutation. Protein model analysis indicated that the Lys232Thr mutation did not disrupt the native network of hydrogen bonds, but has changed the mutual electrostatic forces, resulting in increased instability of the protein.</p><p><b>CONCLUSION</b>The heterozygous Lys232Thr mutation identified in the FGG gene probably underlies the hypofibrinogenemia in this pedigree.</p>

Identification of a novel heterozygous mutation in a pedigree with hereditary coagulation factor XII deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential mutation underlying hereditary coagulation factor XII (FXII) deficiency in a pedigree and explore its molecular pathogenesis.</p><p><b>METHODS</b>Activated partial thromboplastin time (APTT), FXII activity (FXII:C) and FXII antigen(FXII:Ag) and other coagulant parameters of the proband and 5 family members were measured. Potential mutations in the 14 exons and intron-exon boundaries of the FXII gene were screened with polymerase chain reaction (PCR) and direct DNA sequencing. Suspected mutations were confirmed with reverse sequencing. Corresponding PCR fragments from other family members were also sequenced.</p><p><b>RESULTS</b>APPT of the proband and his son were significantly prolonged to 121.5 s and 98.5 s, respectively. FXII:C and FXII:Ag of the proband and his son have reduced to 5%, 6.8% and 9%, 12.2%, respectively. Plasma plasminogen activity (PLG:A) in both individuals was slightly higher than the normal reference range. FXII:C of his second daughter and grandson were slightly reduced to 64% and 60%. FXII:C of the other family members were all in the normal range (72%-113%). A heterozygous missense mutation, g.8597G>A, was identified in exon 13 of the FXII gene in the proband, which resulted in an p.Asp538Asn substitution. For the promoter regions of the FXII gene, the genotype of the proband was 46TT. The same mutations and 46T/T were also found in the proband's son but not in other members of the family. The genotypes of the proband's spouse, eldest daughter and grandson were 46CT, and his second daughter was 46TT.</p><p><b>CONCLUSION</b>The heterozygous mutation of g.8597G>A identified in exon 13 of FXII gene is a novel mutation. Heterozygous p.Asp538Asn mutation and 46TT in the FXII gene can cause hereditary FXII deficiency, which was probably responsible for the low FXII concentrations in this pedigree.</p>

Phenotype and genotype analysis for a consanguineous pedigree with combined coagulation factor VII and X deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential mutations and explore the molecular mechanism underlying combined inherited coagulation factors VII(FVII) and X(FX) deficiency for a family featuring consanguineous marriage between maternal cousins.</p><p><b>METHODS</b>Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FVII activity (FVII:C), FX activity (FX:C), FVII antigen (FVII:Ag), FX antigen (FX:Ag) and other coagulant parameters of the proband and 5 family members were measured. Potential mutations in exons, exon-intron boundaries and 5', 3' untranslated sequences of F7 and F10 genes were screened by polymerase chain reaction and direct sequencing. Suspected mutations were confirmed by sequencing the opposite strand.</p><p><b>RESULTS</b>PT and APTT of the proband were obviously prolonged to become 76.4 s and 60.2 s, respectively. FVII:C, FVII:Ag,FX:C and FX:Ag of the proband were obviously reduced to become 4%, 6%, 6% and 33%, respectively. Both PT and APTT of her grandmother, father, mother and daughter were slightly prolonged, which have measured 16.4 s, 15.8 s,16.9 s, 16.5 s, and 44.0 s, 42.1 s, 41.1 s, 43.5 s, respectively. And their FVII:C (34%, 39%, 31%, 40%, respectively), FX:C (50%, 58%, 47%, 42%, respectively) and FX:Ag (51%, 54%, 58%, 47%, respectively) were slightly reduced, while FVII:Ag was in the normal range. The coagulant parameters of her younger brother were within normal range. Two homozygous mutations, g.11267C to T in exon 8 of F7 gene, which resulted in an Arg277Cys substitution, and g.28139G to T in exon 8 of F10 gene which led to a Val384Phe substitution, were identified in the proband. The proband's grandmother, parents and daughter were heterozygous for both Arg277Cys and Val384Phe mutationss. Wild-type alleles of both F7 and F10 genes were also found in the younger brother.</p><p><b>CONCLUSION</b>A homozygous Arg277Cys mutation and a Val384Phe mutation have been respectively identified in the F7 and F10 genes, which can explain the low levels of FVII and FX in this family. The former has been inherited from the consanguineous parents.</p>

Relationship between the valine/leucine247 polymorphism of apolipoprotein H gene and generation of antiphospholipid antibodies in patients of Han nationality with systemic lupus erythematosus in Wenzhou region / 中华皮肤科杂志

Objective To study the relationship between the valine/leucine247 (817G/T) polymorphism in exon 7 of apolipoprotein H (apoH) gene and the generation of antiphospholipid (APL)antibodies in patients of Han nationality with systemic lupus erythematosus (SLE) in Wenzhou region.Methods This study included 165 patients with SLE and 160 healthy controls of Han nationality in Wenzhou region.Venous blood samples were obtained from all of the subjects followed by the isolation of blood plasma,sera and white blood cells.PCR and DNA sequencing were carried out to assess the Leu/Va1247 polymorphism in apoH gene.Lupus anticoagulant (LAC) was detected by Russell viper venom time (RVVT) assay.Enzyme-linked immunosorbent assay (ELISA) was carried out to quantify the serum levels of anti-β2-glycoprotein Ⅰ (GPI) antibodies and anticardiolipin antibodies (ACA).Chi-square test was carried out to compare the 817G/T polymorphism between the patients and controls,and Logistic regression analysis to evaluate the correlation between the 817G/T polymorphism and production of antiphospholipid antibodies.Results There were significant differences between the patients and controls in the genotype distribution and allele frequency at position 817 of apoH gene (both P ＜ 0.01).The TT,GT genotypes and T allele were more frequent,while GG genotype and G allele were less frequent,in the patients than in the controls.The GT genotype at position 817 was a risk factor for the production of LAC (P＜ 0.05,OR =2.33,95％CI =1.18-4.59),anti-β2GPl antibodies(P＜ 0.01,OR =5.92,95％CI =2.61-13.46) and ACA(P＜ 0.05,OR =2.52,95％CI =1.22-5.24),and the TT genotype was associated with an increased frequency of anti-β2GPI antibodies (P ＜ 0.01,OR =5.84,95％CI =1.69-20.20).Conclusions The 817G/T(Leu/Va1247) polymorphism in exon 7 of apoH gene is associated with the generation of APL antibodies in patients of Han nationality with SLE in Wenzhou region.The TT and GT genotypes at position 817 of apoH gene appear to be a risk factor for the production of APL antibodies.

Identification of a novel mutation of factor XII gene in a family with coagulation FXII deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze genetic mutation and molecular pathogenesis in a family affected with inherited coagulation factor XII(FXII) deficiency.</p><p><b>METHODS</b>Activated partial thromboplastin time (APTT), FXII procoagulant activity (FXII:C), FXII antigen (FXII:Ag) and other coagulants were measured. For affected members of the family, exons 1-14 and flanking intronic regions of the FXII gene were amplified with polymerase chain reaction (PCR) and sequenced thereafter. Expression plasmids containing mutant FXII cDNA was constructed and transfected into COS7 cells transiently. Expressions of FXII:Ag and FXII:C were analyzed.</p><p><b>RESULTS</b>The proband has manifested a prolonged APTT of 108.1 s (reference range: 27.0-41.0 s). Her husband has a normal APTT. Other members of the family had slightly increased APTT. The FXII:C and FXII:Ag of the proband have both dropped to about 0.01 (reference range: 0.72-1.13). The FXII:C levels of her husband, son, daughter and grandchild were 0.57, 0.24, 0.14, 0.16, respectively. And the FXII:Ag levels in her husband, son, daughter and grandchild were 0.55, 0.27, 0.15, 0.21, respectively. The proband and her daughter have both carried a heterozygous deletional mutation 6800-6808delAGCTGGGAG (6800-6808del9bp) in exon 9. For the promoter region of the FXII gene, the genotypes of the proband, her son, daughter and grandchild was TT, whilst that of her husband was CT. Expression study has shown that, whilst the mutant FXII protein has accumulated in the cells similar to wild-type protein, its secretion has reduced approximately by half.</p><p><b>CONCLUSION</b>A novel deletional mutation 6800-6808del9bp has been identified in the FXII gene. Although mutant FXII protein can still accumulate in cells, its secretion has become insufficient. The 6800-6808del9bp mutation and 46T/T have both contributed to the pathogenesis of FXII deficiency in the family, but may have not been the sole cause.</p>

Surgical anatomy and occurrence of injury in choledocho-pancreatico-duodenal junction / 中华消化外科杂志

The choledocho-pancreatico-duodenal junc-tion is located at the central part of choledocho-pancreatico-duodenal region. During early embryogenetic stage, the primary duodenum develops from the end of foregut and the beginning part of the midgut. The dorsal pancreas, hepatic diverticulum and the ventral pancreas which arises from the basic part of hepatic diverticulum are growing and rotating following the duodenum. During the course, the formations of the choledocho-pancreatico-duodenal region and the central part of choledocho-pancreatico-duodenal junction are complete. The injuries in cho-ledocho-pancreatico-duedenal junction may be caused by metal probe or lithotomy forceps for exploring, dilatating the distal bile duct or taking out the stones from the bile duct. Even if the injuries of choledocho-pancreatico-duodenal junction are deve-loped in a limited scope of several centimeters, several adjacent organs may be involved. Injuries in choledocho-pancreatico-duo-denal junction are hard to be identified during operation and may develop into serious pathological procedures.

Mupirocin resistance in Staphylococcus aureus / 中华临床感染病杂志

Objective To investigate mupiroein resistance in Staphylococcus aureus (SAU) and the resistance to commonly used antibiotics in mupirocin-resistant strains. Methods Four hundred and ninety clinically isolated SAU strains froin January 2005 to May 2007 in the First Affiliated Hospital,Wenzhou Medical College were screened by mupirocin(5μg)disc diffusion method.Minimum inhibition concentration(MIC)and the amplification of mupA gene were performed to determine the resistance to mupirocin.Resistance to cefoxitin,gentamycin, levofloxacin, trimethoprim/sulfamethoxazole, rifampin, erythromycin, clindamycin, tetracycline and vancomycin in mupirocin-resistant strains was detected by disc diffusion method, and the amplification of mecA gene was performed to confirm the methieillin resistance among mupiroein-resistant strains.Results Twenty-seven mupirocin-resistant strains were obtained,in which 22(81.5%)were hish-level mupirocin resistant(MuH)and the rest were low-level mupirocin resistant(MuL).Among 27 mupirocin-resistant strains,24 were methicillin-resistant Staphylococcus aureus (MRSA)in which 21 were MuH and 3 were MuL strains.Drug sensitivity tests showed that the resistance to gentamycin,levofloxacin,trimethoprim/sulfamethoxazole,rifampin,erythromycin,elindamycin and tetracycline were hish among MuH and MuL strains,and most of these strains were multi-drug resistant.All strains were susceptible to vaneomycin.Conclusions Most of the clinical emerged mupirocin-resistant SAU strains are MuH and show hish resistance to commonly used antibiotics.Therefore,detection and drug sensitivity test of mupirocin-resistant strains should be strengthened in clinic practice in order to prevent it from dissemination.