Clinical characteristics and genetic analysis of three pediatric patients with idiopathic restrictive cardiomyopathy / 中华心血管病杂志

<p><b>OBJECTIVE</b>Restrictive cardiomyopathy (RCM) is rare in children, and little is known about the molecular basis of RCM. The aim of this study was to investigate the clinical and myopathological characteristics and to detect mutations on cardiac sarcomere protein genes in three idiopathic pediatric RCMs.</p><p><b>METHODS</b>Detailed clinical characteristics and familiar history were obtained in three idiopathic pediatric RCMs. One hundred healthy pediatric individuals were recruited as controls. Histological evaluation was performed with heart tissue retrieved at catheterization in case-1 and case-2. The entire coding sequences of four cardiac sarcomere protein genes, including cardiac troponin T (TNNT2), cardiac troponin I(TNNI3), β-myosin heavy chain (MYH7), and α-actin (ACTC)were screened for mutations. Sequence variants were then tested in the family as well as in 100 healthy control DNAs.</p><p><b>RESULTS</b>All three index cases were diagnosed as primary RCMs without family history, and their clinical conditions deteriorated rapidly. Case-1 was in combination with ventricular septal defect. Case-2 was in combination with mid- and inferoseptal hypertrophy. In case-1, myocardial biopsies displayed extensive an isomorphism and disarray of cardiomyocytes; electron microscopy showed large stacks of severely dysmorphic megamitochondria and focal Z-disc streaming. In case-2, endomyocardial biopsy revealed moderate myocyte hypertrophy with mild interstitial fibrosis; transmission electron microscopy showed misalignment of Z-bands and unequal Z-Z band distances. Genetic analysis identified two heterozygous missense mutations in TNNI3, with R204H in case-1 and R192H in case-3 respectively. A de novo heterozygous deletion in TNNT2 (p. Asn100_Glu101del) was identified in case-2. Sequence analysis shows that all three mutations are located in a position highly conserved across many species. The three mutations were negative for their parents and controls.</p><p><b>CONCLUSION</b>The clinical conditions in all three index cases are deteriorated rapidly after diagnosed as primary RCM. Three heterozygous mutations including two in TNNI3 and one in TNNT2 gene are identified in the three RCMs respectively, which are considered as causative mutations. These findings provide new insights into the molecular etiology responsible for pediatric RCM.</p>

Risk factors for acute fulminant myocarditis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the risk factors for fulminant myocarditis by analyzing clinical symptoms/signs or laboratory findings in children with viral myocarditis.</p><p><b>METHODS</b>The medical data of 71 children with acute viral myocarditis from March 2005 to September 2008 were retrospectively studied. They were classified into fulminant (n=16) and non-fulminant myocarditis groups (n=55). Chi-square and Student's t-test were used to analyze the clinical presentations, laboratory data, EEG and cardiac ultrasound findings on admission. The multiple regression analysis was used to identify the independent risk factors for fulminant myocarditis.</p><p><b>RESULTS</b>Eight children (50%) died in the fulminant myocarditis group, but none in the non-fulminant group. The following factors were closely related to the fulminant course of myocarditis: lower blood pressure, higher serum CK-MB level, positive cTnI, complete atrioventricular block and left bundle branch block, ST segment alterations, prolonged QRS complex, and decreased left ventricular ejection fraction and short axis fractional shortening. Multiple regression analysis revealed that prolonged QRS complex (OR=1.139; CI=1.014-1.279, P<0.05) and decreased left ventricular ejection fraction (OR=0.711; CI=0.533-0.949, P<0.05) were independent risk factors for fulminant myocarditis.</p><p><b>CONCLUSIONS</b>The mortality of fulminant myocarditis is high in children. Prolonged QRS complex and decreased left ventricular ejection fraction on admission are independent risk factors for fulminant myocarditis in children.</p>

Association of Toll-Like Receptor 4 and CD_(14) Gene Polymorphisms with Kawasaki Disease Susceptibility / 实用儿科临床杂志

Objective To explore the association of Toll-like receptor 4 TLR4 and lipopolysaccharide receptor CD14 gene polymorphisms with Kawasaki disease (KD) susceptibility.Methods Three-color fluorescent staining flow-cytometry was used to detect the expression of TLR4 in peripheral blood white blood cell of 76 KD children and 118 healthy control group.The gene of TLR4 (-896A/G), (-1196C/T) and CD14 (-260C/T) polymorphisms was identified by polymerase chain reaction-restriction fragment length polymorphisms; and the relationship between genotype and KD was analyzed.Results 1.The values of mean fluorescence intensity (MFI) of TLR4 in peripheral blood white blood cell of the KD groups and the healthy control groups were 2.87?0.96, 10.55?4.87, 23.36?8.28 and 3.26?0.65, 7.55?1.21, 25.41?6.97, respectively; There was a gradual increase of these values on lymphocyte, neutrophilic leukocyte and mononuclear cell in both groups.2.(-896A/G), (-1196C/T) polymorphisms of TLR4 gene were not found in both groups.3.The frequency of each genotype of CD14 gene (-260C/T) was 35.5%CC, 30.3%CT, 34.2%TT in KD group and 38.1%CC, 47.5%CT, 14.4%TT in healthy control group.The frequency of each genotype was significantly different in 2 groups(?2=11.62 P

Temporal and Spatial Expression Pattern of Tenascin in Zebrafish Early Development / 实用儿科临床杂志

Objective To explore the temporal and spatial expression pattern of tenascin-c(tnc) and tenascin-w(tnw) in zebrafish early development,to further explore the role of tenacsin in zebrafish embryo development,and the association between them.Methods Zebrafish embryos at 2 hours post fertilization(hpf),4 hpt,8 hpt,10 hpf,24 hpt,48 hpr,72 hpf and 7 days post fertilization(dpf) were collected to extract RNA for reverse transcription-polymerase chain reaction(RT-PCR) and fix the embryos at different stages for in situ hybridization.Temporal and spatial expression pattern of tnc and tnw on different stages of zebrafish early development was observed.Results tnc and tnw all expressed in zebrafish from 24 hpf to 7 dpf,but did not expressed from 2 hpf to 10 hpf.Tnc expressed at pharyngeal arch,notochord,somite in 24 hpf,then weakly expressed at somite,but highly expressed at otic vesicle,pectoral fin and hindbrain in 48 hpf,and tnc was expressed at hindbrain,pharyngeal and notochord and disappeared at somite and pectoral.tnw expressed at hindbrain,midbrain and otic vesicle in 24 hpf,expressed at somite,notochord,hindbrain,otic vesicle and pharyngeal in 48 hpf.In 72 hpf,tnw expressed weakly at somite and notochord.Conclusions Zebrafish tnc and tnw have special temporal and spatial expression pattern,and share partial overlapping expression pattern.

Percutaneous Closure of Patent Ductus Arteriosus on Plasma N-Terminal Brain Natriuretic Peptide Level and Left Ventricular Function / 实用儿科临床杂志

Objective To evaluate the effect of percutaneous closure of patent ductus arteriosus(PDA)on left ventricular size and function by measuring plasma N-terminal brain natriuretic peptide(NT-proBNP)level and using two-dimensional echocardiography.Methods According to the modified Ross score,55 children with PDA were divided into 3 groups,no congestive heart failure(CHF)group(31 cases),mild CHF group(14 cases)and moderate-severe CHF group(10 cases).Fifteen age-matched and weight-matched normal children were used as controls.Plasma NT-proBNP was measured using enzyme-linked immunosorbent assay(ELISA).All patients had complete echocardiographic study,including measurement of left ventricular end diastolic volume index(LVEDVI),left ventricular end systolic volume index(LVESVI),and left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS).The correlation between plasma NT-proBNP level and echocardiographic cardiac functional indexes was determined.Results 1.Before operation LVEDVI and LVESVI in PDA patients were obviously higher than those in normal controls(Pa0.05).3.Three months after operation plasma NT-proBNP level,LVEDVI and LVESVI were significantly decreased(P

Matrix metalloproteinase-1 expression in the circulation of patients with Kawasaki disease and its role in the pathogenesis of coronary artery lesions / 中华儿科杂志

<p><b>OBJECTIVE</b>Kawasaki disease (KD) is an acute and self-limited systemic vasculitis syndrome of unknown origin that mainly affects small and medium-sized arteries, particularly the coronary arteries, which is followed by aneurysm formation. Increased levels of matrix metalloproteinase-1 (MMP-1) have been detected in aortic aneurysms in adults, suggesting an important role of MMP-1 in arterial wall destruction and resultant aneurysm formation. The aim of this study was to investigate the potential role of MMP-1 in the pathogenesis of coronary artery lesions in patients with KD.</p><p><b>METHODS</b>Forty patients with KD, including 23 patients without coronary artery lesions (CAL) and 17 patients with CAL, as well as age-matched 10 febrile and 10 healthy afebrile controls were studied. The duration of KD was divided into three phases: the acute phase, the subacute phase and the convalescent phase. Enzyme-linked immunosorbent assay was used to detect the protein levels of MMP-1 in the sera. MMP-1 mRNA expression in the circulating leucocytes was studied using reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>Levels of MMP-1 protein in serum and MMP-1 mRNA expression in the leucocytes were significantly elevated at the acute phase in the two groups of KD patients (CAL group: 14.91 +/- 3.88 ng/ml and 0.89 +/- 0.15 ng/ml; NO-CAL group: 11.27 +/- 3.28 ng/ml and 0.77 +/- 0.14, respectively), compared with febrile (7.05 +/- 1.98 ng/ml and 0.45 +/- 0.12 ng/ml, respectively) and afebrile (5.13 +/- 1.20 ng/ml and 0.29 +/- 0.12 ng/ml, respectively) controls (P < 0.01). Furthermore, MMP-1 protein and MMP-1 mRNA levels were significantly higher in KD patients with CAL than in KD patients without CAL (P < 0.05). There was a significantly positive correlation between the serum protein level of MMP-1 at the acute phase of KD and the circulating leucocytes counts (r = 0.750, P < 0.01). The MMP-1 serum protein level and mRNA expression in the leucocytes at the acute phase of the two KD groups decreased obviously from the subacute through the convalescent phases (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>The expression of MMP-1 at the acute phase of KD was significantly elevated, especially in KD patients with CAL. MMP-1 might be involved in the formation of coronary artery lesions and pathogenesis of KD.</p>

Clinical Significance of Serum Cardiac Troponin in Patients with Dilated Cardiomyopathy / 实用儿科临床杂志

Objective To observe the serum cardiac troponin I (cTnI) level and to analysis the relations between the serum cTnI level and prognosis in patients with dilated cardiomyopathy (DCM) and Cardiac Dysfunction.Methods Serum cardiac cTnI level was measured by enzyme-linked immunosorbant assay. Results The serum cTnI level in DCM patients with class IV cardiac function (0.53 ?0.31) ?g/L was significantly higher than in DCM patients with class Ⅲ cardiac function (0.45?0.27) ?g/L.There was significantly difference in serum cTnI levels between DCM patients with class Ⅲ cardiac function and DCM patients with class Ⅱ cardiac function(0.29?0.27) ?g/L.Conclusion The higher serum cTnI level is correlated with the severity of cardiac function and may be useful for evaluating prognosis in patients with DCM.

Differentiation Between Vaccine Strain and Field Isolates of Classical Swine Fever Virus Using Polymerase Chain Reaction and Restriction Test / 微生物学通报

A nest RT-PCR/restriction test has been developed in order to distinguish the lapinised vaccine strain from field isolates of classical swine fever virus. The restriction enzyme cut sites mapping of the major coding sequence of E2 gene lapinised vaccine strain and ShiMen strain of classical swine fever virus have been compared. Ten and sixteen unique restriction markers have been found in the lapinised vaccine strain and ShiMen strain. The restriction enzyme cut sites mapping of the twenty six unique restriction marker in the major coding sequence of E2 gene of 17 classical swine fever field isolates have been analyzed. Only 3 sites (HgaI、Hin8I及Hsp92I) are present in the lapinised vaccine strain sequence. Two pans of nested primers and a criteria of analysis have been designed for HgaI restriction marker site. The tests have been conducted first on the lapinised vaccine strain and ShiMen strain of classical swine fever virus resulting in predicted restrection patterns. Finally, the tests have been applied to 5 field isolates of different gene group analyzed by phylogenetic study. The result showed that only HCLV strain gene can be cut to 2 fragment by Hgal , and ShiMen strain and 5 field isolates cant be cut At the same time the sensitivity and specificity of nest RT-PCR have been tested. The sensitivity is 0. 2MLD. The specific fragment of BDV and BVDV were not obtained by the nest RT-PCR. These results showed that the development of the nest RT-PCR/restriction tests is very important for the control and perish of classical swine fever in china.

RESEARCH ON THE TESTING OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS BY NESTRT-PCR / 微生物学通报

Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.

Significance of Expression of Tenascin-c Gene in Zebrafish Embryo Development Induced by Ethanol / 实用儿科临床杂志

Objective To explore the expression pattern of tenascin-c(tnc)gene in zebrafish embryo abnormal development which was induced by ethanol,and to further understand the function of tnc gene in embryo develepment.Methods Zebrafish were treated with ethanol at different concentration from 100 to 500 mmol/L,and embryos at 24 and 48 hours were collected and fixed,then tnc expression pattern was observed by in situ hybridization and reverse transcriptase-polymerase chain reaction(RT-PCR).Results The result of RT-PCR showed that ethanol at 100 and 200 mmol/L could increase the expression of tnc,while the result of in situ hybridization showed that,while ethanol at 300 mmol/L and above decrease the expression of tnc in presumptive position at 24 hours,and ethanol at 100 mmol/L and above caused increase expression of tnc in zebrafish heart.Conclusions tnc is increased when treated with 100 and 200 mmol/L ethanol and is presented in the abnormal development of hearts of zebrafish,which can promote the normal development of embryos in some degrees.The expression pattern of tnc in pathologic state is highly conserved in all vertebrate,and in adult and embryos as well.