OBJECTIVE: To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling. METHOD: Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan. RESULT: One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan. CONCLUSION Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.
Adolescent ; Child ; Child, Preschool ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Oligonucleotide Array Sequence Analysis ; Sulfate Transporters

Objective　To explore the relationship between hemolymph phenol oxidase and the melanization of Plasmodium yoelii oocysts in Anopheles dirus. Methods　An Anopheles dirus-Plasmodium yoelii system was used Anopheles dirus were divided into 3 groups, that is, non-blood-fedding (N), normal-blood-fedding (B) and infected-blood-fedding (I). The activities of MPO and o-DPO in hemolymph from 3 groups were determined with native polyacrylamide gel electrophoresis (PAGE) and density scanning at 5, 7, 11 and 15 d after blood feeding. Results　Both MPO and o-DPO activity were significantly higher in group I than group N and B (P<0.05). But with the melanization of Plasmodium yoelii oocysts, both MPO and o-DPO activity in group I were decreased in comparison with group N, especially on the 15 th day after infected-blood feeding. MPO and o-DPO activity in group B were significantly stronger than those of group N. Conclusion　Blood feeding and infection of Plasmodium yoelii both can activate the cascade. The heamolymph phenol oxidase may play an important role in the melanization of Plasmodium yoelii oocysts in Anopheles dirus.

OBJECTIVETo examine the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1, VEGFR2) in transdifferentiated human proximal tubular epithelial (HK-2) cell induced by transforming growth factor beta1 (TGFbeta1).

METHODSThe transdifferentiation of HK-2 cells was detected by evaluation of expression of alpha-SMA by cytoimmunochemistry and RT-PCR. The VEGF mRNA was evaluated with RT-PCR. The secreted VEGF in the culture media was measured with ELISA. The cellular VEGF, VEGFR1, and VEGFR2 were measured with Western blot.

RESULTSThe immunostain of alpha-SMA were positive in HK-2 cell induced by TGFbeta1 at the concentration of 5 and 8 ng/ml for 72 h. The expression of alpha-SMA mRNA was induced by TGFbeta1 in concentration- and time-dependent manners. The expressions of mRNA and protein of VEGF were upregulated by TGFbeta1 at the concentration of 0.1 and 1 ng/ml for 72 h and at the concentration of 8 ng/ml for 12 h and 24 h when compared with the control. But expressions of mRNA and protein of VEGF were downregulated by TGFbeta1 at the concentration of 3, 5, and 8 ng/ml for 72 h and at the concentration of 8 ng/ml for 36, 48, and 72 h, respectively. Meanwhile, Protein levels of VEGFR1 and VEGFR2 were upregulated by TGFbeta1 in concentration- and time- dependent manners.

CONCLUSIONSIncreased expression of VEGFR1 and VEGFR2 and two-phase change in VEGF expression occurred in the process of tubular epithelial transdifferentiation induced by TGFbeta1. Reduced expression of VEGF may contribute to tubular epithelial transdifferentiation in a vicious circle.

Cell Differentiation ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules, Proximal ; cytology ; RNA, Messenger ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1 ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Objective To explore the relationship of height,weight and body mass index with fists size among adults,and to compare the regression coefficient of different models.Methods Adults aged 20 -80 years were selected by stratified cluster sampling methods.T -test and univariate analysis of variance were used to compare the height,weight and fists size stratified by gender and BMI group.Regression model for fists volume was established.The regression coefficients were compared through covariance interaction analysis.Results The height,weight,BMI and fists size of male were higher than that of female (P <0.01 ).The regression model for fists size(Y)of BMI(X)for male,female and total population were Y=1 67.603 +9.445X,Y =1 1 1 .1 83 +7.779X and Y =77.1 57 +1 1 .028X(R2 =0.324,0.322,0.271 ,P <0.05).Fists size of different BMI groups were related to their height (R2 =0.501 ,0.432,0.624,0.692,P <0.05),respectively, and their regression coefficients have no statistically differences (P =0.07).The fists sizes of different BMI groups were different(P <0.01 ).Fists size of overweight or obesity groups (365.641 ±68.51 4,365.641 ±68.51 4,respectively)were larger than that of the normal and slim groups(P <0.01 ),and fists size of normal body (31 4.261 ±59.590)was larger than that of slim body(261 .081 ±59.478).Conclusion The fists sizes of male were higher than that of female.Fists size was positively related to their BMI.The height had the some impact on fists size among people in different BMI groups.

OBJECTIVE: To explore the relationship between the time-dependent level changes of microRNA and 18S rRNA and the different postmortem interval (PMI) in rat cardiac muscle. METHODS: SD rats were sacrificed by cervical dislocation and placed at ambient temperature 25 degrees C with a humidity of 50%. Total RNA was extracted from the rat cardiac muscle at different time points after death. The levels of miR-1-2 and 18S rRNA were examined using real-time PCR in rat cardiac muscle. The results were expressed by cycle threshold (Ct) value to explore relationship between PMI and Ct value, and the regression functions were established to estimate PMI. RESULTS: The miR-1-2 level in rat myocardial tissue showed no significant changes within 120 h after death, and then began to decline. The 18S rRNA level increased gradually within 96 h after death, and then declined slowly. The nonlinear relationships were established between Ct value (18S rRNA), deltaCt value (difference between 18S rRNA and miR-1-2) and PMI. The R2 of conics fitting were 0.9487 and 0.8072, respectively. CONCLUSION Ct value of 18S rRNA and deltaCt value present a good correlation with PMI, and can be markers for estimating early PMI.
Animals ; Forensic Pathology ; Male ; MicroRNAs/metabolism* ; Myocardium/pathology* ; Postmortem Changes ; RNA Stability ; RNA, Messenger/metabolism* ; RNA, Ribosomal, 18S/metabolism* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction/methods* ; Time Factors

This study is to investigate the effects of phenytoin sodium, lidocaine (sodium channel blockers), propranolol (beta-adrenergic receptor antagonist), amiodarone (drugs prolonging the action potential duration) and verapamil (calcium channel blockers) on arrhythmia of mice induced by Bufonis Venenum (Chansu) and isolated mouse hearts lethal dose of Chansu. Arrhythmia of mice were induced by Chansu and then electrocardiograms (ECGs) were recorded. The changes of P-R interval, QRS complex, Q-T interval, T wave amplitude, heart rate (HR) were observed. Moreover, arrhythmia rate, survival rate and arrhythmia score were counted. Isolated mouse hearts were prefused, and the lethal dose of Chansu was recorded. Compared with control group, after pretreatment with phenytoin sodium, broadening of QRS complex and HR were inhibited, and the incidence of ventricular arrhythmia was reduced dramatically, while survival rate was improved; the isolated mouse hearts lethal dose of Chansu was increased significantly. After pretreatment with lidocaine, the prolongation of P-R interval and broadening of QRS complex were inhibited, and the incidences of ventricular arrhythmia were reduced dramatically, while survival rate was improved; the isolated mouse hearts lethal dose of Chansu was increased significantly. After pretreatment with propranolol, prolongation of P-R interval, broadening of QRS complex, prolongation of Q-T interval and HR were inhibited, and the incidences of both supraventricular and ventricular arrhythmias were reduced dramatically, while survival rate was improved. After pretreatment with amiodarone, HR was inhibited, the incidences of ventricular tachycardia were reduced dramatically. Lastly, after pretreatment with verapamil, the prolongation of P-R interval and Q-T interval were inhibited and the incidences of both supraventricular and ventricular arrhythmias were reduced dramatically; the isolated mouse hearts lethal dose of Chansu was reduced significantly. In in vivo experiments, phenytoin sodium was most effective against the mice arrhythmias induced by Chansu while cautious use of verapamil for Chansu inducing arrhythmia should be noted. It is also concluded that mice ventricular arrhythmias induced by Chansu might be most closely related to sodium channel, supraventricular arrhythmias might be related to beta-adrenergic receptor, and calcium channel plays an important role in conduction block. In in vitro experiments, phenytoin sodium was most effective, followed by lidocaine and propranolol, and amiodarone had no obvious effect and verapamil reduced the lethal dose of Chansu.
Amiodarone ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Arrhythmias, Cardiac ; chemically induced ; physiopathology ; Bufanolides ; toxicity ; Electrocardiography ; drug effects ; Female ; Heart Rate ; drug effects ; In Vitro Techniques ; Lethal Dose 50 ; Lidocaine ; pharmacology ; Male ; Mice ; Phenytoin ; pharmacology ; Propranolol ; pharmacology ; Verapamil ; pharmacology

OBJECTIVETo evaluate the safety, effectiveness, and outcomes of holmium laser enucleation of the prostate (HoLEP) for patients with symptomatic enlarged prostate after 11 years of experience.

METHODSThe 3162 evaluable patients treated with holmium laser enucleation of the prostate at our institution between August 2001 and August 2011 were retrospectively analyzed. Study variables included International Prostate Symptom Score, quality of life, maximum urinary flow rate, and incidence of complications.

RESULTSHoLEP were performed successfully completed, not patients which occurs as electric cutting syndrome. The operation time was (60.8 ± 18.4) minutes; average resection of prostate quality was (45.4 ± 24.4) g. The hemoglobin reduce though surgery was (1.81 ± 0.93) g/L; percentage of red blood cell change was 1.24% ± 0.43%, and sodium blood drop was (1.14 ± 0.35) mmol/L. Postoperative patients of hospital stay (3.1 ± 1.1) days, average time of indwelling catheter time was (2.3 ± 0.8) days. Patients were followed up for 6-131 months time, an average of 32.4 months. Postoperative patients with international prostate symptom score progressive declined. The quality of life score was 2.2 ± 1.7, and it less than preoperative (5.7 ± 3.3, t = 2.447, P < 0.01). The time of follow-up droped further, and postoperative comparative differences have statistical significance (t = 2.179, 2.228, 2.306 and 2.365, P < 0.05). The maximum urinary flow rate also improved (P < 0.05). Postoperative complications included bladder neck contracture (4 cases), urinary tract infection (107 cases), urethral stricture (11 cases) and urinary incontinence (11 cases). The 11 patients reoperation.

CONCLUSIONSHoLEP treatment of benign prostatic hyperplasia could achieve the advantages of open surgery the same effect. It had fewer damage, faster recovery, fewer complications, and is a good treatment option.

Adult ; Aged ; Aged, 80 and over ; Humans ; Lasers, Solid-State ; Male ; Middle Aged ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome

BACKGROUNDH3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.

METHODSIn order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).

RESULTSIn this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.

CONCLUSIONThe results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.

Animals ; COS Cells ; Cercopithecus aethiops ; Dogs ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; Plasmids ; Reassortant Viruses ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, Attenuated ; immunology ; Viral Proteins ; genetics

OBJECTIVETo investigate the association between rs751141 gene polymorphisms in EPHX2 gene and essential hypertension in Kazak and Han in Xinjiang.

METHODSA total of 267 essential hypertensive patients in Kazaks, 368 essential hypertensive patients in Hans, 284 normotensive controls in Kazaks and 348 normotensive controls in Hans were enrolled in this study. TaqMan assay was used to detect the rs751141 G/A gene polymorphisms of EPHX2 gene.

RESULTSThe rs751141 G/A genotype frequencies for GA + AA genotypes was 40.2 percent in essential hypertensive subjects and 52.0 percent in control subjects in Hans, respectively. The genotype frequencies were significant difference between the two groups in Hans in Xinjiang (P < 0.01). The rs751141G/A gene polymorphism had no significant difference between essential hypertensive patients and normotensive controls in Kazaks in Xinjiang (P > 0.05).

CONCLUSIONThe essential hypertension in Kazaks in Xinjiang is not associated with rs751141G/A gene polymorphism of EPHX2 gene, but the essential hypertension in Hans in Xinjiang is associated with rs751141G/A allele gene polymorphism of EPHX2 gene. A type of rs751141 allele gene polymorphism may be the independent protective factor of essential hypertension in Hans in Xinjiang.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; epidemiology ; Epoxide Hydrolases ; genetics ; Gene Frequency ; Genotype ; Humans ; Hypertension ; epidemiology ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide

In order to explore the feasibility of inducing the human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) to differentiate into insulin-secreting cells with biological products alone, hUC-MSCs were separated and purified from the whole umbilical cord by the sequent digestion of collagenase II and trypsin followed by two-step centrifugation. hUC-MSCs were induced with IMDM culture medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and Ginkgo biloba extract (GBE). Before and after the induction, the morphological changes were observed under inverse microscope; the islet-related genes were detected by RT-PCR; islet-like clusters (ILCs) were identified by dithizone (DTZ) staining; PDX-1 and immunoreactive insulin (IRI) were examined by immunofluorescence method; the quantity and quality of IRI secretion were assayed by chemiluminescence immunoassay and Western blot respectively. The results showed that the purified hUC-MSCs presented long spindle-like shape and parallel or spiral arrangement which are typical morphological features of MSCs. After the induction, hUC-MSCs changed gradually into round or oval shape and gathered together to form ILCs; there were more than one hundred clusters on the growth surface of a flask of T25; ILCs were stained into positive mauve by DTZ and positive for PDX-1 and IRI; Western blot displayed that most of the IRI was proinsulin (PI). Therefore, hUC-MSCs can rapidly differentiate into insulin-secreting cells under the sole induction of EGF, bFGF, GBE and IMDM, but ILCs are not mature enough to produce sufficient true insulin.
Cell Differentiation ; Cells, Cultured ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Ginkgo biloba ; chemistry ; Humans ; Insulin-Secreting Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Plant Extracts ; pharmacology ; Umbilical Cord ; cytology