OBJECTIVETo investigate genetic etiology of fetal urinary abnormalities with array-based comparative genomic hycridization(array-CGH).

METHODSThirty-two fetuses with variable urinary abnormalities but normal karyotyping by conventional cytogenetic technique were selected. DNA from the fetuses and their parents samples were prepared and hybridization with Affymetrix cytogenetic 2.7M arrays by follwing the manufacture's standard protocol. The data were analyzed by special CHAS software packages.

RESULTSBy using array-CGH detection, genomic imbalanced copy number variations (CNVs) were identified in night fetuses(28%), four out of night CNVs were inherited from parental samples; two were indicated to be benign variants(6%) in the database; and the other three CNVs (9%) were all de novo adjacent microdeletions and microduplication mapping on to common chromosome 1q21.1 region, within which was genitourinaty system function associated gene PDZK1.

CONCLUSIONThe incidence of genomic unbalanced variations in fetuses with congenital urinary malformations is approximately 28%, including about 9% pathogenic variations. Copy number variations (CNVs) of chromosome 1q21.1 region are associated with congenital urinary malformations which may be due to haploinsufficiency or overexpression of PDZK1 gene.

Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Comparative Genomic Hybridization ; DNA Copy Number Variations ; Female ; Humans ; Kidney ; abnormalities ; Pregnancy ; Prenatal Diagnosis

Objective To investigate the concentration of β-amyloid peptide 42 (Aβ42) in cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) and its first clinical event-clinically isolated syndrome (CIS), and explore its associations with duration, disability severity and total T2-hyperintense lesion numbers in MRI. Methods Thirty-three patients with MS, 23 patients with CIS and 13 controls were investigated in this study. The disability severity of patients with MS and CIS in attack period was assessed by Expanded Disability Status Scale (EDSS). MRI scanning of brain, spinal cord or optic nerve was performed. And Aβ42 concentration in CSF was assessed by liquid chip assay. Results No significant differences of Aβ42 concentrations in CSF from patients with MS and CIS in attack period were noted as compared with those from controls ([104.78±13.73]pg/mL, [134.13±25.06] pg/mL vs. [137.02±23.35]pg/mL, P>0.05). ButAβ42 concentration in CSF from patients with secondary progressive MS (SPMS, [167.99±36.39]pg/mL) was significantly higher than that from patients with relapsing-remitting MS (RRMS, [92.74±13.64] pg/mL, P=0.042). No correlations of Aβ42 concentration in CSF with the duration of MS and CIS and scores of EDSS were noted in patients with MS and CIS (P> 0.05). The concentration of Aβ42 in CSF from patients with MS with a duration for more than one year lower than the ones with a duration for less than one year, but the difference was not significant (P>0.05). Total T2-hyperintense lesion numbers in MRI of patients with MS and CIS were positively correlated with Aβ42 concentration in CSF (MS patients: r=0.507, P=0.038; CIS patients:r=0.485,P=0.049). Aβ42 concentration in CSF from patients with MS with total T2-hyperintense lesions ≥4 (129.34±19.96) was significantly higher than that from the ones with total T2-hyperintense lesions <4 (73.51±12.60, P=0.049). Conclusion Axonal damage in patients with SPMS is more severe than that in patients with RRMS.Increased CSF Aβ42-level in patients with MS is a feature of disease progression. There is a possible relation between T2-hyperintense lesion load and axonal damage in patients with MS.

OBJECTIVETo clone human bone morphogenetic protein-2 (hBMP2) gene and construct its eukaryotic expression vector pcDNA3.1 -hBMP2.

METHODSHuman BMP2 gene was amplified by RT-PCR method from human osteosarcoma cells and constructed into eukaryotic expression vector pcDNA3.1-hBMP2. The gene in the vector pcDNA3.1-hBMP2 was identified by PCR amplification, enzyme digestion and DNA sequencing.

RESULTSThe cloned DNA was confirmed to be hBMP-2 gene.

CONCLUSIONIn this study, hBMP2 gene is successfully cloned and its eukaryotic expression vector pcDNA3.1-hBMP2 is constructed, which provides the foundation of using BMP2 gene therapy to accelerate new bone formation in distraction osteogenesis.

Bone Morphogenetic Protein 2 ; Cloning, Organism ; Genetic Therapy ; Genetic Vectors ; Humans ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Transfection

OBJECTIVETo observe the effect of Cornus Officinalis total glycosides (COTG) and Cornus polysaccharides (CP) on myocardial mitochondria and expression levels of glycogen synthase kinase-3β (GSK-3β) of acute myocardial infarction (AMI) rats.

METHODSThe AMI rat model was established by ligating the left anterior descending branch of coronary artery. Rats were divided into 5 groups according to random digit table, i.e., the sham-operation group, the model group, the COTG prevention group, the CP treatment group, the COTG treatment group, 12 in each group. Normal saline was administered to rats in the normal control group and the model group by gastrogavage. Corresponding medication was respectively administered to rats in the rest 3 groups by gastrogavage. The cardiac function was detected by echocardiography and hemodynamics. The infarct size was determined by Masson trichrome staining. The expression of mitochondrial biogenesis genes such as a subunit of peroxisome proliferators-activated receptor-γ coactivator-1 (PGC-1α), PGC-1β, nuclear respiratory factor-1 (NRF-1), and GSK-3P mRNA were detected by Real-time PCR.

RESULTSCompared with the sham-operation group, the myocardial infarction size increased, cardiac function decreased, the expression of PGC-1α, PGC-1β, and NRF-1 mRNA decreased, and the expression of GSK-3β mRNA increased (all P <0. 05). Compared with the model group, myocardial infarction sizes were reduced, cardiac function was improved, the expression of NRF-1 mRNA was elevated in the COTG prevention group, the CP treatment group, the COTG treatment group; the expression of the PGC-1α and PGC-1β mRNA was elevated in the COTG prevention group and the CP treatment group; the expression of GSK-3β mRNA was reduced in the CP treatment group (all P <0. 05). Compared with the CP prevention group, fractional shortening (FS) and aortic systolic blood pressure (SBP) increased in the CP treatment group; ejection fraction (EF) decreased in the CP treatment group; the expression of PGC-1α, PGC-1β, NRF-1 mRNA were reduced in the the CP treatment group and the COTG treatment group; the expression of GSK-3β mRNA decreased in the CP treatment group (all P <0. 05). Compared with the COTG treatment group, FS, EF, left ventricular end systolic pressure (LVESP), SBP, and the expression of GSK-3β mRNA were reduced in the CP treatment group (P <0. 05).

CONCLUSIONSCOTG and CP could improve cardiac function, reduce the myocardial infarction area, and promote biogenesis of myocardial mitochondria. Their protective effects on the mitochondria of cadiocytes might be achieved by GSK-3β signalina pathway.

Animals ; Cornus ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Glycosides ; Heat-Shock Proteins ; Mitochondria, Heart ; physiology ; Myocardial Infarction ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Polysaccharides ; Protective Agents ; pharmacology ; therapeutic use ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Transcription Factors

Objective The clinical data of dilated cardiomyopathy (DCM) patients with or without pulmonary hypertension (PH) diagnosed by echocardiography were compared.Methods During January 2007 to December 2009,61 cases of DCM with PH and 51 cases of DCM without PH were admitted in our department.The demographic and clinical data,heart function,echocardiography and serum total bilirubin and creatinine levels of all patients were analyzed.Results Sex,age,vital signs,combined diseases and arrhythmias as well as the serum creatinine level [ ( 103.5 ± 49.7 ) μmol/L vs.( 90.3 ± 37.3 ) μmol/L,P ＞0.05] were similar between the two groups,while the incidence of NYHA Ⅲ and IV (95％ vs.65％ ),the left ventricle end-systolic dimension[ (71.0 ±9.6) mm vs.(65.5 ±7.2) mm],dimension of the left atrium [ (52.8 ±8.93) mm vs.(43.9 ±6.3) mm],right ventricular outflow tract [ (29.1 ±5.3) mm vs.(22.1 ±3.3) mm] incidence of pericardial effusion (29/61 vs.7/51 ) and the serum total bilirubin level [ (45.3 ±31.8) μmol/L vs.(19.5 ±9.08) μmol/L] were significantly higher while ejective fraction was significantly lower in DCM with PH than those in DCM without PH (0.28 ±0.10 vs.0.36 ±0.10,all P ＜0.05 ).Conclusion DCM patients with PH is linked with worse clinical features than DCM patients without PH.

Background Infantile hemangioma (IH) is the most common benign tumor in children with prevalence in the face and neck.Various treatment options including oral propranolol have been described for IH,but the mechanism of drugs remains enigmatic.The aim of this study was to investigate the pathogenesis and establish a reliable in vivo model of IH which can provide platform for drug exploration.Methods Stem cells from the proliferating hemangiomas (HemSCs) were isolated by CD133-tagged immunomagnetic beads.Their phenotype and angiogenic property were investigated by flow cytometry,culturing on Matrigel,real-time polymerase chain reaction (PCR),immunofluorescent staining and injection into BALB/c-nu mice.Results HemSCs had robust ability of proliferating and cloning.The time of cells doubling in proliferative phase was 16 hours.Flow cytometry showed that HemSCs expressed mesenchymal markers CD29,CD44,but not endothelial/hematopoietic marker of CD34 and hematopoietic marker CD45.The expression of CD105 was much lower than that of the reported hemangioma derived or normal mesenchymal stem cell (MSC).Real-time PCR showed that the mRNA levels of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF) and matrix metalloproteinase-1 (MMP-1) of HemSCs were higher than that of neonatal human dermal fibroblasts (NHDFs) and human umbilical vein endothelial cells (HUVECs).After HemSCs were cultured on Matrigel in vitro,they formed tube-like structure in a short time (16 hours) and differentiated into endothelial cells in 7 days.After 1-2 weeks of implantation into immunodeficient mice,HemSCs generated glucose transporter 1 positive blood vessels.When co-injected with HUVECs,the vascularization of HemSCs was greatly enhanced.However,the single implantation of HUVECs hardly formed blood vessels in BALB/c-nu mice (P ＜0.05).Conclusions HemSCs may be some kinds of primitive mesoderm derived stem cells with powerful angiogenic ability,which can recapitulate human hemangioma by co-injecting into immunodeficient mice with HUVECs.

OBJECTIVETo investigate the effect of bilateral mandibular distraction osteogenesis in the condyles.

METHODS16 adult hybrid dogs were randomly divided into normal control group and experiment group. Experimental dogs underwent bilateral mandibular osteodistraction at a rate of 1 min/day. 4 dogs were killed respectively in distraction period, 2 and 8 weeks after completion of 10 days distraction. The bilateral condyles specimens were harvested and examined with histological and immunohistochemical methods.

RESULTSCompared with normal control group, various degrees of irregularities and erosion were found in fibrocartilage of condyle in experiment group, including damage in fibrous layer, hyperplasia layer and proliferative layer and osteogenic activity in cartilage layer. A significant increase of TGF-beta1 expression was also found in experiment groups. TGF-beta1 positive staining was noted in hypertrophic cell, matrix and chondroblast, osteoblast and matrix in osteogenic activity areas. These changes were the most obvious in 2 weeks after completion of distraction.

CONCLUSIONGradual bilateral mandibular distraction at a rate of 1 mm/day brought degenerative changes of condyle, but the changes are reversible.

Animals ; Dogs ; Mandible ; Osteoblasts ; Osteogenesis, Distraction ; Temporomandibular Joint ; Transforming Growth Factor beta1

OBJECTIVEIn contrast to CD(4)(+) helper T-lymphocytes (T(H)), little is known about the transcriptional regulation of CD(8)(+) cytotoxic T-lymphocytes (Tc) and its role in the pathogenesis of asthma is unclear. This study was conducted to investigate the effect of T-bet and GATA-3 mRNA expression on profiles of type 1 and type 2 cytotoxic T lymphocytes in asthmatic children.

METHODTotally 38 asthmatic children, including acute attack group composed of 20 cases (age 3 - 13 years, mean 6.2 +/- 2.9), remission group with 18 cases (age 3 - 12 years, mean 6.1 +/- 2.5) and 20 healthy control children (age 3 - 12, 6.9 +/- 2.7) were recruited in this study from Sep. 2005 to Mar. 2006. The mRNA expression of T-bet and GATA-3 in the peripheral blood mononuclear cells were detected by using semi-quantitative PCR and Tc1, Tc2 cell numbers by flow cytometry analysis system.

RESULTT-bet mRNA in asthmatic children was lower than that in control group and lower in attack stage than in remission stage (0.14 +/- 0.04, 0.21 +/- 0.03, 0.28 +/- 0.03, P < 0.05). In contrast, GATA-3 mRNA was higher in asthmatic children than in control group and higher in attack stage than in remission stage (0.49 +/- 0.09, 0.44 +/- 0.08, 0.37 +/- 0.04, P < 0.05). It was shown that Tc1 percentage was lower in asthmatic children than those of control group and lower in attack stage than those of remission stage (6.6 +/- 2.4, 14.2 +/- 4.3, 31.2 +/- 3.8, P < 0.05). Tc2 percentage in asthmatic children was higher than that of control group and higher in attack stage than that of remission stage (10.0 +/- 4.2, 5.4 +/- 2.2, 3.5 +/- 1.1, P < 0.05). Spearman correlation analysis revealed that T-bet mRNA was positively correlated with Tc1 percentage (r = 0.704) and negatively correlated with Tc2 percentage (r = -0.629). GATA3 mRNA was negatively correlated with Tc1 percentage (r = -0.612) and positively correlated with Tc2 percentage (r = 0.673). The T-bet/GATA-3 mRNA ratio was positively correlated with Tc1 percentage (r = 0.731) and Tc1/Tc2 (r = 0.773), while negatively correlated with Tc2 percentage (r = -0.642).

CONCLUSIONThe imbalance of T-bet/GATA-3 mRNA expression is closely correlated with skewed Tc2 dominance in asthmatic children.

Adolescent ; Asthma ; genetics ; immunology ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Male ; RNA, Messenger ; genetics ; metabolism ; T-Box Domain Proteins ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the risk factors of the serious complications related with double-J ureteral stent placement following percutaneous nephrolithotomy (PCNL).

METHODSClinical data were reviewed for 272 patients treated with PCNL and indwelling double-J stents between January, 2014 and April, 2016. The risk factors of serious complications were identified using univariate and multivariate logistic regression analysis.

RESULTSSerious complications of double-J ureteral stenting occurred in 63 patients (23.1%). Univariate and multivariate logistic regression analysis indicated that the ureter abnormalities (β=1.735, P=0.000, OR=5.670), stent indwelling duration (β=1.206, P=0.028, OR=3.340), gender (β=0.895, P=0.016, OR=2.446), preoperative urinary tract infection (β=0.849, P=0.020 , OR=2.338) and stent size (β=0.847, P=0.011, OR=2.333) were all risk factors of serious complications related with the procedure.

CONCLUSIONMale patients are exposed to a higher risk of serious complications following PCNL. Effective management of urinary tract infection and choice of appropriate stent size in cases of ureteral abnormalities help to reduce these complications. The double-J stent should be withdrawn as soon as possible in patients with good postoperative recovery.

Female ; Humans ; Kidney Pelvis ; Logistic Models ; Male ; Nephrostomy, Percutaneous ; Postoperative Period ; Risk Factors ; Stents ; adverse effects ; Ureter ; surgery ; Ureteral Obstruction ; surgery

OBJECTIVETo detect the expression of lung aquaporin 5 (AQP5) in mice with acute allergic asthma and the effect of dexamethasone (DEX) treatment on AQP5 expression, and investigate the role of AOP5 in asthma pathogenesis.

METHODSMouse models of acute allergic asthma were randomly divided into acute asthma group, normal control group and DEX treatment group. The total number of white blood cells, the subpopulations, and the levels of IL-5 and IFN-gamma were detected in the bronchoalveolar larvage fluid (BALF). The lung tissue AQP5 mRNA expression was detected by RT-PCR, and AQP5 distribution by immunohistochemical method.

RESULTSIn asthma group, the total white blood cells, eosinophils and IL-5 levels were all significantly higher (P<0.01) and IFN-gamma levels lower than those of the control group (P<0.01). After DEX treatment, the levels underwent a significant reverse change (P<0.05, P<0.01, P<0.01, and P<0.01, respectively). AQP5 mRNA expression in the asthma group was significantly higher than that in the control group (P<0.01), and was significantly lowered with DEX treatment (P<0.01). Extensive inflammatory changes, mucus hypersecrection, several edema and inflammatory cell infitration around the blood vessels were observed in the lung tissue of the mice in the asthma group. The morphological changes of the treatment group were significantly ameliorated. AQP5 protein was detected in the type I alveolar epithelial cells, the airway columnar epithelial cells and the apical membranes of the submucosal gland acinar cells in the control group. Stronger AQP5 protein expression was found in the asthma group.

CONCLUSIONAQP5 is over-expressed in mice with acute asthma which is possibly associated with mucus hypersecrection. DEX can inhibit AQP5 expression and ameliorate allergic airway inflammation, edema and mucus hypersecrection.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Aquaporin 5 ; biosynthesis ; genetics ; Asthma ; genetics ; metabolism ; prevention & control ; Dexamethasone ; pharmacology ; Female ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction