Objective To investigate the characteristics of family environment in children with attention deficit hyperactivity disorder (ADHD) in clinics, and analyse the risk factors for ADHD. Methods Two thousand two hundred and ninety-six children with inattention, hyperactivity or unfavourable school performance were subjected to diagnosis with DSM-Ⅳ criteria in clinics. The characteristics of family environment were investigated by self-prepared questionnaires. The risk factors for ADHD were explored by univariate analysis and noneonditioned multivariate Logistic regression analysis. Results Seven hundred and twenty children were diagnosed with ADHD. There were significant differences in family environment between children with ADHD and those without(P<0.05 or P<0.01). The risk factors for ADHD included discord between parents, parental smoking and maternal depression during pregnancy and after delivery, while older age, female, paternal higher educational background were protective factors for ADHD. Conclusion Unfavourable family environment may be associated with the prevalence of ADHD, and special attention should be paid to the family environment in the treatment of ADHD.

Cell viability of Pichia pastoris was detected by flow cytometry (FCM) with two reagents fluorescein diacetate (FDA) and propidium iodide (PI). Compared with FDA/PI double-stained dot plots and PI single-stained dot plots,the latter could divide dead and living cells into two separate zones,and get the correct proportion. Then PI single-stained method was used to detect the change of cell viability in Pichia patoris fermentation. At glycerol batch and fed-batch phase,little dead cells were detected. At methanol fed-batch phase,cell viability decreased when cell weight increased,and was only 73.8% at 88 h.

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.

The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.

Objective:To find a short, neutralizing antibody-inducible, ORF2-encoded protein by means of comparing the immunogenicity of pN472-C617 and pN477-C613 which represent amino acids 472-617 and 477-613 of HEV ORF2-encoded protein of hepatitis E virus(HEV) genotype 4, respectively.Methods:The two recombinant proteins were expressed, purified, and then used to immunize BALB/c mice. Anti-HEV titers in the immune sera were detected by ELISA. Anti-HEV neutralizing activity was tested by a PCR-based in vitro neutralization assay.Results:Both of the two recombinant proteins were efficiently expressed in E.coli in soluble forms. The purified proteins induced mice to develop high levels of anti-HEV specific antibodies. However, only the immune sera obtained from the mice immunized with pN472-C617 showed the neutralizing activity to the homologous HEV strain by preventing the virus from absorption on PLC/PRF/5 cells surfaces and replication in the cells. The immune sera against pN477-C613, which was truncated five amino acids from both N- and C-terminal of pN472-C617, had no HEV neutralizing activity.Conclusion:The pN472-C617 is the shortest neutralizing antibody-inducible ORF2-encoded protein of HEV reported in literatures so far. It may be considered as a potential candidate for a novel HEV subunit vaccine in our future study.

Objective To explore the improvement of core symptoms and detailed function in children with attention deficit hyperactivity disorder (ADHD) after treatment with methylphenidate extended-release tablets, and analyse the relationship between core symptoms reduction and detailed function improvement. Methods One hundred and fifty-six children with confirmed ADHD were rated with Swanson, Nolan, and Pelham, Version Ⅳ (SNAP-Ⅳ)Scale before treatment, then methylphenidate extended-release tablets were orally administered 18 mg once daily for 1 month, and children were rated again by means of SNAP-Ⅳ Scale and detailed function improvement questionnaire. The core symptoms reduction and detailed function improvement were observed, and their relationship was analysed. Results The primary mean scores of each factor in SNAP-Ⅳ Scale decreased significantly after treatment with methylphenidate extended-release tablets(P< 0.001). The reduction rate of factor IOWA/I/O was the most significant (>50%), followed by ADHD-H/Im and ADHD-In. The performance of school study, homework doing and social behavioral function was improved, and the detailed function was significantly improved. The reduction rate in ADHD-In factor was significantly correlated with improvement of school study and homework doing (P<0.01). The reduction rate in ADHD-H/Im factor was significantly correlated with improvement of social behavioral function(P<0.05). Conclusion Methylphenidate extended-release tablets play a role in both core symptoms reduction and detailed function improvement in children with ADHD, and core symptoms reduction is related to detailed function improvement to some degree. Methylphenidate extended-release tablets exert different effects on different detailed function.

Glucose was transported by the large number of hexose transporters in yeast cells. There were 18 hexose transporter genes had been identified in Saccharomyces cerevisiae. However,as an excellent expression system,there was no information of these genes had been reported in Pichia pastoris. Based on high homologous recombination efficiency in yeast,we chose G418 resistance for screening,200 bp were cloned from the up and down sequences of HXT1 ORF respectively,then ligated to the 5′ and 3′ end of G418 resis-tance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates,finally we obtained one HXT1 gene deletion mutant named GS115?HXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.

OBJECTIVEThis study aimed to investigate the value of child oral health for Chengdu parents, their intentions, and factors influencing their decision to acquire oral insurance coverage for their childrens.

METHODSA total of 562 Chengdu parents were interviewed using questionnaires by convenient sampling, and the results were analyzed using SPSS 20.0.

RESULTSThe age of children (B = -1.741, P = 0.004), age of parents (B = 2.031, P = 0.003), level of oral discomfort (B = 0.569, P = 0.000), incurring/not incurring oral care expenses in the previous year (B = 1.897, P = 0.014), the last time parents' had teeth cleaned (B = 0.777, P = 0.006), and acquiring/not acquiring commercial insurance coverage (B = 1.632, P = 0.031) significantly influenced the intention of acquiring child oral insurance.

CONCLUSIONChild oral health, health and insurance awareness of parents, and other factors influenced the intention of parents to purchase oral insurance coverage for their children, which were significant to establish pediatric dental insurance.

Child ; China ; Dental Care ; Health Expenditures ; Humans ; Insurance, Dental ; economics ; Oral Health ; economics ; Parents ; Surveys and Questionnaires