ABSTRACT
To analyze the mutations of F12 genein one pedigree with congenital factor FXII (FXII) deficiency , and investigatethe molecular mechanisms of FXII deficiency . Methods Activated partial thromboplastin time(APTT),Prothrombin time(PT), FXII activity(FXII:C), FXII antigen(FXII:Ag) and other coagulant parameters were tested in the proband and his family members .5'and 3'UTR,all exons and their exon-intron boundaries of F12 gene were analyzed by direct sequencing .The detected mutations were confirmed by reverse sequencing .100 healthy persons were as normal controls .Results The proband showed a markedly prolonged APTT (106.4s), the FXII:C and FXII:Ag were 2.0% and 1.0%, respectively .Hissecond daughter and granddaughter had slightly prolonged APTT , and other family members are normal.The FXII:C and FXII:Ag of family members were also decreased ( his son, 23.0% and 21. 0%;his elder daughter , 23.0%and 23.0%;his second daughter ,24.0%and 23.0%;hisgranddaughter , 23.0%and 23.0%).The phenotype of all members is consistent with cross -reactive material negative . Nucleotide sequencing analysis showed that the proband had missense mutations in the F 12 gene, including one homozygous mutationc.1556T >G ( p.Leu519Arg) and a commonly reported single nucleotide polymorphism site within the promoter region of the F 12 gene (46T/T) .Sequencing results from the proband 'children demonstrate them as carriers of a heterozygous missense mutation .The proband 's wife is normal and with 46C/C in the promoter region .Conclusion The c.1556T>G in exon 13 is a novel mutation .This mutation affects FXIIcatalytic function , associated with a reduced level of FXII .
ABSTRACT
To control and prevent the Echinococcus in the place ,we established a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Echinococcus species specific DNA from dog faeces .Four primers which recognizing 6 dis-tinct regions on the NADH dehydrogenase subunit 2 (ND2) gene of Echinococcus granulosus were designed and used for LAMP assay .The specificity of LAMP assay was evaluated using DNA extracted from Echinococcus granulosus , Taenia saginata , and other dog intestinal parasites .In addition ,the sensitivity of LAMP assay was compared with that of conventional PCR using recombinant plasmid carrying Echinococcus granulosus ND2 gene fragment as standard template DNA after 10-fold serial dilution .Furthermore ,we extracted DNA from 46 canine fecal samples collected from endemic areas ,and tested the copro-DNA samples using LAMP and necropsy method .Results showed that E .g ND2 primer sets could differentiate Echinococcus granulosus from Echinococcus multilocularis without cross reaction among other parasites detected .Furthermore ,the LAMP assay with primer sets to the ND2 gene could detect 4 × 101 copies of target gene ,demonstrating 103 times higher sensitivity than that of conventional PCR methods .The LAMP assay with primer set to ND2 gene showed good sensitivity and specificity to detect copro-DNA samples extracted from fecal samples of 46 dogs tested in endemic areas .There was no statistically signifi-cant difference among LAMP and necropsy .In this study ,a sensitive ,specific and rapid copro-DNA detection LAMP assay was developed successfully for diagnosis of dogs infected with Echinococcus granulosus .Due to its rapidity ,simplicity ,speci-ficity and sensitivity ,the LAMP assay is a promising new tool for rapid detection of dogs infected with Echinococcus spp .dur-ing the field survey or in poor-equipped laboratories .
ABSTRACT
Objective To explore the changes and clinical significance of serum human cartilage glycoprotein-39(HC gP39),osteopontin (OPN) and rheumatoid factor (RF) in patients with rheumatoid arthritis (RA).Methods Serum HC gP39,OPN levels were measured by ELISA in 98 patients with RA and 98 healthy controls.Serum RF was detected by nephelometric immunoassay.Results The serum HC gP39,OPN and RF levels of RA group were significantly higher than the healthy control group (t =20.25,32.71,36.34,all P ＜ 0.01),and serum HC gP39,OPN and RF levels in active phase patients were higher than the inactive phase patients (t =24.22,45.62,50.15,all P ＜0.01).The levels of serum HC gP39,OPN increased gradually with the increase of the staging severity(F =18.48,12.36,all P ＜0.05).The serum level of RF was positively correlated with the levels of HC gP39,OPN in patients with RA (r =0.682,0.656,all P ＜ 0.01),the serum level of HC gP39 was positively correlated with the level of OPN (r =0.608,P ＜ 0.01).Conclusion The HC gP39,OPN and RF reflect situation of RA patients and have close relationship with the clinical progression of RA,which can be used as one of important indexes to judge RA activity and different staging.
ABSTRACT
Objective To comprehend etiology and clinical manifestation changes of infant pneumonia in this locality.Methods Indirect immunofluorescence (IIF) assay was applied in children with acute pneumonia to detect serum 11 kinds of viruses[respiratory syncytial virus (RSV),adenovirus (ADV),influenza virus (IFV-A+B),parain fluenza virus(PIV14) ,coxsackie B,virus(CB1V),Coxsackie A7 virus (CA7V) ,ECHO virus]specific antibody IgM,according to the serum virus-specific IgM positive,C-reactive protein(CRP)<8mg/L and no other pathogenic infection and laboratory evidence for the conditions of 436 cases detected in children with pneumonia.Results Detected a total 125 cases of antibody-positive,the positive detection rate is 37.99%.Of which 103 cases of single virus infection .accounting for 82.4% ,22 cases of mixed infection,accounting for 17.6%.RSV infection on top of the list followed by the rest of IFV,ADV and PIV.Infants of different ages,different seasons of the different types of virus susceptibility.Conclusion Pneumonia in infants were caused by pathogenic bacteria in addition to the virus of a wide range,and the incidence of age,the peak seasons and the clinical manifestations were vary.From an early stage of infection pathogen detection,clearing pathogen type,making the correct diagnosis of pneumonia in the treatment of infants had an important guiding significance.