Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

OBJECTIVESTry to induce liver-derived dendritic cells proliferation by plasmid-IL-3 and CD40L genes in mice in vivo.

METHODSRapid tail-vein injection of large amounts of plasmid-carrying genes was performed to transfect genes in mice livers. Liver nonparenchymal cells were isolated by Percoll gradient centrifugation. Dendritic cell markers were detected and dendritic cells were sorted out by flow cytometry. Morphology of dendritic cells was studied microscopically (with Giemsa staining) and under scanning electron microscopy.

RESULTSLiver nonparenchymal cells dramatically increased in the liver lobules, portal and periportal areas in the treated group, but not in the control group. B220+/DEC205+ dendritic cells were detected and sorted by flow cytometry. There were more B220+/DEC205+ dendritic cells (16.0%) in the experiment group than those in the control group (1.1%). Morphologically, the sorted B220+/DEC205+ cells showed irregular shaped nuclei, paucity of cytoplasmic granules and extensive cytoplasmic processes.

CONCLUSIONB220+/DEC205+ dendritic cells were expanded in vivo in mice livers by rapid tail-vein injection of plasmid-carrying genes encoding IL-3 and CD40L in a large amount. Inducing liver dendritic cell proliferation in vivo provides a more convenient way for studying the biology of these cells.

Animals ; CD40 Ligand ; genetics ; Dendritic Cells ; cytology ; Flow Cytometry ; Hepatocytes ; cytology ; Interleukin-3 ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Transfection

Objective To explore the molecular basis of familial hypercholesteraemia(FH)by analyzing the phenotype and genotype relationship through identify the low density liporotein receptor(LDL-r)gene mutation in a FH kindred.Methods A male patient of 15 years old was selected to examine the electrocardiogram,lipid.Color Doppler was used to examine heart and great vessels.The promoter region and the 18 exons of the LDL-r gene were screened by touch-down polymerase chain reaction(PCR)and DNA sequencing.Results The caro-tid intima-media thickness(IMT)was increased to 0.23 cm,while coronary flow velocity reserve(CFVR)was decreased to 1.57,and mode-rate mitral regurgitation was found in the proband.The genetic alteration G→A change at 1 448 of exon 10 causing premature stop codon(W462X).The same heterozygous nonsense mutation was also found in his father.The mutation had been reported in other Chinese patients.In vitro experiments showed that W462X mutation leads to low LDL binding and internalization ability.Conclusions The homozygous mutation(W462X)in exon 10 of the LDL-r gene were identified in the clinically heterozygous FH proband.The W462X mutation is the underl-ying cause of hypercholesterolaemia and clinical AS manifestations.W462X is recurrent mutation among Chinese FH patients.It might be a hot spot mutation in LDL-r in Chinese FH.J Appl Clin Pediatr,2009,24(1):18-20

OBJECTIVETo analyze the correlation between myocardial ischemia and carotid atherosclerosis in hypertensive patients.

METHODSThe clinical data were collected from 85 hospitalized hypertensive patients admitted between May 2005 and September 2008 without the complication of coronary artery disease as confirmed by cardiac computed tomographic angiography (CTA). According to the results of treadmill exercise test, the patients were divided myocardial ischemia group and ischemia-free group. Univariate and multivariate analyses were used to screen the risk factors of myocardial ischemia. The correlations were analyzed between myocardial ischemia, common carotid artery intima-media thickness (IMT), Crouse score of the carotid plaque, thickness of the intraventricular septum and left artrium. The receiver operating characteristic (ROC) curves were used to evaluate the sensitivity and specificity of IMT and Crouse score in predicting the presence of myocardial ischemia in hypertensive patients.

RESULTSCarotid plaque formation was identified as the major risk factor of myocardial ischemia in hypertensive patients (OR=4.982, P=0.004). The incidence of myocardial ischemia in the hypertensive patients with carotid plaques was significantly higher than that in the patients without the plaque (Chi2=9.317, P=0.002). Myocardial ischemia in hypertensive patients was positively correlated to the thickness of the intraventricular septum (r=0.362, P=0.001) and left artrium (r=0.298, P=0.009), and the IMT of the common carotid artery was positively correlated to the thickness of the intraventricular septum (r=0.231, P=0.045). The area under cure (AUC) of the ROC curve of Crouse score was 0.726-/+0.061 in predicting the presence of myocardial ischemia in the hypertensive patients (P=0.001), and that of IMT was 0.682-/+0.061 (P=0.006).

CONCLUSIONCarotid plaque formation is the major risk factor of myocardial ischemia in hypertensive patients and shows a positive correlation to the onset of myocardial ischemia, but both the common carotid artery IMT and the Crouse score of the carotid plaque are not accurate markers for predicting myocardial ischemia in patients with hypertension.

Adult ; Aged ; Aged, 80 and over ; Atherosclerosis ; complications ; pathology ; Carotid Artery, Common ; diagnostic imaging ; pathology ; Coronary Angiography ; methods ; Female ; Humans ; Hypertension ; complications ; pathology ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Ischemia ; etiology ; pathology ; Risk Factors ; Tomography, X-Ray Computed ; Ultrasonography

BACKGROUND: In orthodontics, micro-implant anchorage in the infrazygomatic crest that cannot damage the tooth root can achieve an unobstructed overall movement of the upper dentition. However, little is reported on the stress and strain of the tooth and alveolar bone during the distal movement of the upper dentition. OBJECTIVE: To set up a three-dimensional finite element model to perform a biomechanical analysis of micro-implant anchorage in the infrazygomatic crest for the distal displacement of the upper dentition at different heights. METHODS: Cone-beam CT data from a female patient admitted for orthodontic treatment was saved in Dicom format, and imported into Mimics 16.01 software. Then, a three-dimensional model of the right maxilla and tooth dentition was made by automatically and manually selecting boundaries. The model was imported into Geomagic8.0 for removal of noise dots and smooth processing, and then it was imported into the Mimics16.01 software and meshed for the surface/body through 3 Matics software. Afterwards, three-dimensional models maxillary denture, archwires and traction hooks and implants were established by ProE5.0, and all the models were imported into ANSYS13.0 and assembled and analyzed for stress and strain analysis. RESULTS AND CONCLUSION: We successfully established the three-dimensional finite element model for biomechanical analysis of micro-implant anchorage in the infrazygomatic crest for the distal displacement of the upper dentition at different heights, and this model conformed to the anatomic features. With the increase of the height of traction hooks (1, 4, 7, 10 mm), the vertical stress of the maxillary teeth increased gradually, and had no correlation with the change of the horizontal stress. With the increase of the height of traction hooks, at the sagittal axis, the strain at midpoints of middle incisors, canine teeth, and first molars decreased gradually and the strain at the root of middle incisors and canine teeth also decreased gradually, but there was no change in the strain at the root of first molars. With the increase of the height of traction hooks, at the vertical axis, the strain at the midpoints and tooth root of middle incisors increased, while the strain of canine crown increased gradually and that of the canine root decreased; the strain at the midpoint of first molars changed a little, and the strain of the tooth root decreased gradually. The dentition rotated from clockwise to counterclockwise. To conclude, the three-dimensional finite element model made in the study is consistent with the anatomic structure, which provides a basis for biomechanical analysis of micro-implant anchorage in the infrazygomatic crest for the distal displacement of the upper dentition. The upper dentition impedance center located in the position of 4 to 7 mm of the arch wire can be used as the microimplant support site in the infrazygomatic crest.

Objective To explore the Electrocardiography (ECG) changes of residents in Keshan disease area and the status of growth and decline of Keshan disease in Shaanxi Province. Methods Using stratified randomized sampling method,2 mild,2 moderate and 2 serious disease counties were selected respectively in 2005 and 2006. A total of 6 counties were sampled,2 villages,one with severe disease and one with mild,were selected from each sampled county. A total of 12 villages were selected. The clinical examination and ECG were conducted in 3-year old children of agricultural population of the selected villages. Results ECG of 5692 cases were performed in the selected 12 village in the 6 counties,in which 4917 cases showed normal electrocardiogram,up to 86.38% (4917/5692). Two hundred and fifty-two cases showed roughly normal electrocardiograms,up to 4.43%(252/5692). Five hundred and twenty-three cases had abnormal electrocardiogram,accounting for 9.19% (523/5692). Among them,the abnormal electrocardiogram rates in mild,moderate and serious disease areas were 7.07% (144/2036), 11.41%(167/1646) and 10.54%(212/2010),respectively. Atrioventricular block was the major abnormal electrocardiogram change,followed by arrhythmia,ST-T changes,and low voltage. One hundred and thirty-nine cases were confirmed as latent and chronic Keshan diseases. One hundred and thirty-one cases were latent Keshan, and detection rate was 2.30%(131/5692). Eight cases were chronic Keshan,and the detective rate was 0.14% (8/5692). Complete right bundle branch block [37.06% (63/170) ],ST-T changes [22.35% (38/170) ],multiple premature ventricular beats [12.94% (22/170)] were the major abnormal electrocardiogram change of Keshan patients. Conclusions Atrioventricular block and arrhythmia are the major abnormal electrocardiogram changes. Keshan disease incidences are controlled under a stable condition.

OBJECTIVETo investigate the pathogen of children with acute respiratory infection (ARI) in Suzhou and to provide some evidences for clinical diagnosis and treatment.

METHODSThe nasopharyngeal secretion samples from 2492 inpatient children with ARI, during the period of November 2005 to May 2007, were investigated for respiratory syncycial virus (RSV), influenza virus A and B, parainfluenza virus type 1, 2, 3 and adenovirus by both the indirect immunofluorescence assay and virus isolation. Human metapneumovirus (hMPV) were examined by reverse-transcription polymerase chain reaction (RT-PCR) at the same time.

RESULTSOf 2492 samples tested, 961 (38.6%) were positive. The total positive rate of virus pathogens in children with ARI was found related to age, season and respiratory disease. The detection rates by age were: 50.0% (412/824), 43.4% (190/438), 30.5% (207/679)and 27.6% (152/551), chi(2) = 96.5002, P < 0.01; the detection rates by season were : 46.7% (366/784), 13.8% (66/478), 13.8% (59/428) and 58.6% (470/802), chi(2) = 392.3279, P < 0.01; the detection rates by disease were (acute upper respiratory infection, acute laryngitis, throat-trachea-bronchitis, bronchial pneumonia, pneumonia genuine, bronchiolitis, bronchial asthma): 21.4% (30/140), 73.7% (14/19), 32.0% (8/25), 36.9% (598/1620), 13.1% (8/61), 66.1% (216/327) and 29.0% (87/300), chi(2) = 162.1276, P < 0.01. There was no association between the total positive rate and sex. The detection rates by sex were: 39.0% (588/1508) for male and 37.9% (373/984) for female, chi(2) = 0.2962, P > 0.05. The peak of RSV appeared from December to March. There was the highest RSV detection rate 50.2% (164/327) with bronchiolitis. The hMPV can be detected all year around. The peak of hMPV appeared in winter and the detection rate was 13.2% (106/802).

CONCLUSIONRSV and hMPV are the main respiratory viral pathogens in Suzhou. Detection of viral pathogens in children with respiratory infection could give fast, accurate diagnostic evidence, and help avoid antibiotics abuse.

Acute Disease ; Adenoviridae ; Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; isolation & purification ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; etiology ; virology ; Sequence Analysis, DNA

OBJECTIVETo determine the value of fluorescence in situ hybridization (FISH) to the diagnosis of chromosome abnormality in genetic diseases and prenatal diagnosis.

METHODSFISH was performed using appropriate probes, including alpha-satellite DNA probe, chromosome sequence specific probe and whole chromosome painting probe, to examine the blood samples from 36 patients who were suspected of having chromosome abnormality by conventional cytogenetics, and to examine the amniocytes from 45 pregnant women who were in need of prenatal diagnosis.

RESULTSAmong 36 patients, the following karyotypes 45, X; 45, X/46, XX; 45, X/46, Xr(X); 46, X, i(Xq); 47, XXY; 46, XX, t(4;7); 47, XYY; 47, XXX; 47, XXY, inv(7); 46, XY, inv(7); 47, XX, +21 were detected by FISH. Of the fetuses of the 45 pregnant women, two fetuses with chromosomal abnormalities were diagnosed by FISH; the karyotypes were 47, XX, +18 and 46, XY, der(15) t(Y;15) respectively.

CONCLUSIONFISH can precisely and rapidly detect the chromosome abnormalities. It is a complement to the conventional cytogenetics and can be widely used in the diagnosis of genetic diseases and prenatal diagnosis.

Adult ; Amniocentesis ; Chromosome Aberrations ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Male ; Prenatal Diagnosis ; Sex Chromosome Aberrations ; Turner Syndrome ; diagnosis

OBJECTIVETo investigate the effect of local application of bFGF combined with sucralfate on the cell proliferation during continuous tissue expansion (CTE).

METHODSNine white pigs were selected to undergo the continuous tissue expansion in this study and treated with bFGF and sucralfate, respectively as the following groups: group 1 with both bFGF and sucralfate, group 2 only with bFGF, group 3 with only sucralfate, and group 4 with saline as control. Fifteen samples were taken in each pig for immunohistochemistry analysis 1-14 days and 6 weeks after the operation.

RESULTSIn the group with both bFGF and sucralfate, the epidermic basal cells proliferated significantly after the operation and reached top level in 3 days, which was statistical higher than the control group, but the multiplication of basal cell was the lowest 14 days after the operation, still more than the control group. In dermal layer, proliferation of fibroblasts, vessel endothelial cells, hair follicles epidermic cells and sweat gland epicytes was also significant higher in the group with both bFGF and sucralfate than that the control group and reached top level 7 day after the operation, but the proliferation of cells decreased obviously 14 days after the operation, still higher than the control group. The mitotic activity of cells returned to the basal level in 42 days. There were no significant differences among the group 2, group 3 and group 4.

CONCLUSIONLocal application of both bFGF and sucralfate could be more effect to induce cells multiplication during early skin expansion to facilitate the growth of neoformed skin soft tissue.

Animals ; Cell Proliferation ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Fibroblast Growth Factors ; pharmacology ; Fibroblasts ; cytology ; drug effects ; Sucralfate ; pharmacology ; Swine ; Time Factors ; Tissue Expansion ; Tissue Expansion Devices

AIMTo observe the expression of Presenilin-1 (PS-1) and production of amyloid beta-protein (Abeta) in hippocampus of female senile rats and to investigate the effect of vitamin E(VE) on preventing Alzheimer's disease after menopause.

METHODSThe animal model was established using female senile rats. Experimental groups (n=8) were respectively given different doses of VE(5 mg/kg, 15 mg/kg, 60 mg/kg) per day. The expression of PS-1 in hippocampus was detected by immunohistochemistry, the level of Abeta in hippocampus was measured by Radioimmunoassay, and neuronal ultrastructure in hippocampal DG area was observed using transmission electron microscope.

RESULTSThe expression of PS-1 in rat hippocampus of senile control group was stronger than that of adult control group. PS-1 expressed weakly in three medication groups along with augmentation of dosage. The levels of Abeta were found to correlate statistically with the expression of PS-1. The content of Abeta in VE groups was significantly decreased compared to that in senile control group (P < 0.01). There were some changes in the neuronal ultrastructure of senile rats. Neurons were gradually recovered in VE groups.

CONCLUSIONVE may depress the production of Abeta by regulating the expression of PS-1, reducing neuronal injuries. VE may play a role in neuronal protection.

Aging ; Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Female ; Hippocampus ; drug effects ; metabolism ; Presenilin-1 ; metabolism ; Rats ; Rats, Wistar ; Vitamin E ; pharmacology