Adrenal Hyperplasia, Congenital ; complications ; metabolism ; Body Height ; Growth Disorders ; etiology ; Humans ; Risk Factors ; Steroid 21-Hydroxylase ; metabolism

Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P

OBJECTIVETo evaluate the performance of AB-8 macroporous adsorption resin for adsorption and desorption of flavones in liquorice.

METHODSThe concentration of flavones in liquorice was determined by ultraviolet spectrophotometry, and the adsorption behavior of AB-8 macroporous adsorption resin to flavones in liquorice was examined for the adsorption capacity and the volume of solution loaded.

RESULTSOptimal adsorption of flavones was achieved with the sample pH of 5, total flavones concentration in the solution of 0.85 mg/ml, sample flow velocity of 3 BV/h, and washing with 60% ethanol at the flow velocity of 3 BV/h.

CONCLUSIONAB-8 macroporous adsorption resin can be well applicable for enrichment of flavones in liquorice.

Adsorption ; Flavones ; chemistry ; isolation & purification ; Glycyrrhiza ; chemistry ; Macromolecular Substances ; chemistry ; Porosity ; Resins, Synthetic ; chemistry ; Spectrophotometry, Ultraviolet

AIMTo observe the effects of inulicin on the neointimal hyperplasia and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) after balloon injury in rat aorta.

METHODSThe intimal hyperplasia model was replicated by balloon injury. The histological changes in vascular wall were observed by HE staining. MMP-2 activation was tested by gelatin zymogram analysis; MMP-2 and TIMP-2 expression was detected by Western blot and immunohistochemistry analysis.

RESULTSNeointimal hyperplasia induced by balloon injury was significantly inhibited by inulicin. The proteolytic activity and the expression of MMP-2 and ratio of MMP-2 and TIMP-2 were also returned to control level by inulicin after balloon injury.

CONCLUSIONInulicin inhibits hyperplasia of neointima by modulating the balance of MMP-2 and TIMP-2.

Animals ; Endothelium, Vascular ; drug effects ; enzymology ; metabolism ; Hyperplasia ; prevention & control ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Neointima ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Sesquiterpenes ; pharmacology ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

Objectives To investigate the relationship between single nucleotide polymorphism (SNP)H558R in SCN5A gene and chronic Keshan disease (KSD) complicated with hypertension,and the relationship between H558R and occurrence of arrythmia in chronic KSD complicated with hypertension.MethodsThirty nine patients with chronic KSD complicated with hypertension and 63 geographical region matched hypertension control subjects were recruited in our study in Fuyu county,Qiqihaer city,Heilongjiang province between 2006 and 2010.H558R polymorphism in case and control groups was genotyped using the polymerase chain reaction single-strand conformation polymorphism(PCR-SSCP) and sequenced,and electrocardiography(ECG) characteristics were examined in the two groups.Case-control study analytical methods were applied to analyze the relationship between H558R and chronic KSD complicated with hypertension,and the relationship between H558R and occurrence of arrythmia in chronic KSD patients complicated with hypertension.Results Subjects of genotype 558 TC in the case group had a decreased risk of chronic KSD complicated with hypertension with odds ratio of 0.288[95％ confidence interval (CI):0.104 - 0.794],and subjects of genotype TC in chronic KSD complicated hypertension patients had a decreased risk of QRS prolongation with odds ratio of 0.061 (95％CI:0.006 - 0.612).Conclusions Polymorphism H558R in SCN5A gene may be a predisposition factor of chronic KSD complicated with hypertension and occurrence of arrythmia in chronic KSD complicated with hypertension.

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism

OBJECTIVETo study the clinicopathologic features, immunohistochemical profiles and prognosis of chromophobe renal cell carcinoma (ChRCC).

METHODSForty-two cases of ChRCC were retrieved from the archival files of the Affiliated Hospital of Qingdao University, 401 Hospital of PLA and Qingdao Municipal Hospital from 2003 to 2011. The clinical and pathologic features of the tumors were reviewed. Hale colloidal iron staining was performed and EnVision immunohistochemistry was used to detect the expression of a series of immunologic markers. Forty cases of clear cell renal cell carcinoma and 10 cases of renal oncocytoma were selected as controls.

RESULTSThe patients included 17 males and 25 females. The age of patients ranged from 39 years to 78 years (median age = 57 years). On gross examination, the tumors ranged from 2 cm to 19 cm in greatest dimension (mean size = 7.3 cm). Histologically, the tumors were mainly composed of solid sheets, acini or tubules of malignant cells. The tumor cells contained clear finely reticular ("chromophobe") and eosinophilic cytoplasm with perinuclear clearing. The nuclear outline was irregular and wrinkled. Nucleoli were inconspicuous and mitotic figures were barely seen. Hale colloidal iron stain was positive in all cases. Immunohistochemically, the tumor cells were variably positive for EMA (100%, 42/42), CK7 (95.2%, 40/42), Ksp-cad (92.9%, 39/42), CK18 (88.1%, 37/42), CD117 (61.9, 26/42), CD10 (31.0%, 13/42) and PAX2 (28.6%, 12/42). They were negative for vimentin, CA IX and TFE3. The follow-up period in 31 patients ranged from 2 to 77 months (average duration = 29 months). Three patients died of tumor metastasis 3, 8, 13 months respectively after the operation. Twenty-eight patients were still alive without evidence of tumor recurrence.

CONCLUSIONSChRCC predominantly occurs in middle-aged and elderly patients. It often carries a favorable prognosis. The presence of plant cell-like morphology, pale cells with uniform reticular microvesicular appearance and perinuclear clearing are characteristic histologic features. The diffuse positivity for Hale colloidal iron stain and EMA/CK7/Ksp-cadherin/CD117-positive immunoprofiles are also useful in differential diagnosis.

Adenoma, Oxyphilic ; metabolism ; pathology ; Adult ; Aged ; Cadherins ; metabolism ; Carcinoma, Renal Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Immunophenotyping ; Keratin-18 ; metabolism ; Keratin-7 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Lung Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Nephrectomy ; methods ; Treatment Outcome

OBJECTIVEMany children were found to have low free carnitine level in blood by tandem mass spectrometry technology. In some of the cases the problems occurred secondary to malnutrition, organic acidemia and other fatty acid oxidation metabolic diseases, and some of cases had primary carnitine deficiency (PCD). In the present article, we discuss the diagnosis of PCD and evaluate the efficacy of carnitine in the treatment of PCD.

METHODWe measured the free carnitine (C0) and acylcarnitine levels in the blood of 270 000 neonates from newborns screening program and 12 000 children with suspected clinical inherited metabolic diseases by tandem mass spectrometry. The mutations of carnitine transporter protein were tested to the children with low C0 level and the diagnosis was made. The children with PCD were treated with 100 - 300 mg/kg of carnitine.

RESULTSeventeen children were diagnosed with PCD, 6 from newborn screening program and 11 from clinical patients. Mutations were found in all of them. The average C0 level [(2.9 ± 2.0) µmol/L] in patients was lower than the reference value (10 µmol/L), along with decreased level of different acylcarnitines. The clinical manifestations were diverse. For the 6 patients from newborn screening, 4 were asymptomatic, 1 showed hypoglycaemia and 1 showed movement intolerance from 2 years of age. For the 11 clinical patients, 8 showed hepatomegaly, 7 showed myasthenia, 6 showed cardiomyopathy, 1 showed chronic abdominal pain, and 1 showed restlessness and learning difficulty. Among these patients, 14 cases were treated with carnitine. Their clinical symptoms disappeared 1 to 3 months later. The C0 level in the blood rose to normal, with the average from (4.0 ± 2.7) µmol/L to (20.6 ± 8.3) µmol/L (P < 0.01). However, the level was still lower than the average level of healthy children [(27.1 ± 4.5) µmol/L, P < 0.01].

CONCLUSIONSeventeen patients were diagnosed with PCD by the test levels of free carnitine and acylcarnitines in blood with tandem mass spectrometry, and gene mutation test. Large dose of carnitine had a good effect in treatment of the PCD patients.

Cardiomyopathies ; diagnosis ; drug therapy ; genetics ; Carnitine ; analogs & derivatives ; blood ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Female ; Follow-Up Studies ; Humans ; Hyperammonemia ; diagnosis ; drug therapy ; genetics ; Infant ; Infant, Newborn ; Male ; Muscular Diseases ; diagnosis ; drug therapy ; genetics ; Mutation ; Neonatal Screening ; methods ; Organic Cation Transport Proteins ; deficiency ; genetics ; Reference Values ; Tandem Mass Spectrometry

OBJECTIVESurvey of the coastal city of Yantai, from human and swine hepatitis E virus (HEV) genotype correlation.

METHODApplication of reverse transcription nested polymerase chain reaction (RT-nPCR) method for local acute sporadic hepatitis E patients,normal population of HEV-IgM positive and local pig farm pigs were HEV RNA detection. And HEV RNA positive samples for cloning sequencing and sequence analysis.

RESULTSIn 16 patients with acute sporadic hepatitis E in 7 cases of RNA positive stool specimens of HEV; 51 IgM positive sera of normal people in specimens with 1 HEV RNA positive; 34 pig bile specimens with 1 HEV RNA positive. Sequence analysis revealed the region HEV strains and swine strains in the ORF2 region of nucleotide sequence homology is 87%-98.1%. 7 strains of hepatitis E virus genotype in patients and 1 strains of swine hepatitis E virus genotypes are type IV, gene sequence homology between the 87%-98.1%; there were 6 patients and porcine gene sequence homology in 93.9%-98.1% between,for type a subtype; 1 patients and porcine gene sequence homology in 87%, for the type D subtype. Normal population of 1 cases of hepatitis E virus genotype for I type D subtype. Human and porcine HEV ORF2 gene fragment and HEV part I-IV representative strains were compared, and the nucleotide sequence homology were 82.5%-100%, 81.7%-92.9%, 81.4%-93.9%, 84.9%-100%.

CONCLUSIONThe area population prevalence of HEV in the presence of 2 genotype 3 subtype genes, mainly to IV A, in pigs with popular HEV gene with a high homology; HEV type I in the crowd disperses in the presence of.

Animals ; Genotype ; Hepatitis E ; diagnosis ; epidemiology ; Hepatitis E virus ; classification ; genetics ; Humans ; Phylogeny ; RNA, Viral ; analysis ; Sequence Analysis, DNA ; Swine

AIMTo observe the change of potassium current on cultured neurons differentiated from hippocampus neural stem cells of the newborn rat.

METHODSNeural stem cells from newborn rat hippocampus were cultured in vitro and passaged continuously. Differentiation of the cell was induced by serum and removing mitogens. After differentiation cells were plated on plastic dishes and cultured for 1 d, 7 d, 14 d and 21 d. Whole-cell voltage patch clamp recording was used respectively to detect voltage-dependent K+ current.

RESULTSAfter 1 d culture, no current was detected, and on the 7th d, 14th d, 21st d after differentiation, the amplitude of K+ currents was (18.077 +/- 2.789)pA/pF, (13.099 +/- 2.742)pA/pF, (34.045 +/- 8.067)pA/pF at +50 mV. The recorded K+ current included two components that could be blocked by TEA and 4-AP separately, assumed the slowly inactivating delayed rectifier K+ current (IK) and the fast inactivating transient outward K+ current (IA).

CONCLUSIONThe function of potassium channels on the hippocampus neural stem cells of the newborn rat approaches mature gradually when the time of differentiation becomes longer in vitro.

Animals ; Animals, Newborn ; Cells, Cultured ; Delayed Rectifier Potassium Channels ; physiology ; Hippocampus ; cytology ; Neural Stem Cells ; cytology ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; physiology ; Potassium Channels, Inwardly Rectifying ; physiology ; Rats ; Rats, Sprague-Dawley