This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.

Objective To observe the effect of hydrogen on ultraviolet B (UVB)-induced oxidative damage to skin fibroblasts.Methods Primary human skin fibroblasts from foreskin tissues were divided into five groups:normal control group receiving no treatment,hydrogen control group treated with hydrogen-rich saline,UVB group receiving irradiation only,post-treatment group irradiated with UVB followed by hydrogen-rich saline treatment,and pre-treatment group treated with hydrogen-rich saline followed by UVB irradiation.The dose of UVB was 30,60 and 90 mJ/cm2 in the cell proliferation assay and 90 mJ/cm2 in the other experiments.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of fibroblasts,a chemiluminescence method to estimate the activity of superoxide dismutase (SOD) and catalase as well as to determine the level of malondialdehyde in the culture supernatant of fibroblasts,enzyme linked immunosorbent assay (ELISA) to determine the supernatant level of 8-isoprostane-prostaglandin F2α (8-iso-PGF2α),Western blot to detect the expression of heme oxygenase-1 (HO-1) in fibroblasts.One-factor analysis of variance was conducted to assess differences in these parameters among these groups.Results UVB irradiation decreased the proliferative activity (absorbence value at 490 nm) of fibroblasts in a dose-dependent manner.Both the pre-treatment group and post-treatment group showed a statistical increase in proliferative activity of cells compared with the corresponding UVB control groups (all P ＜ 0.05).The activity of SOD and catalase as well as the protein expression of HO-1 were significantly higher (all P ＜ 0.05),whereas the supernatant levels of malondialdehyde and 8-iso-PGF2α were statistically lower (both P ＜ 0.05) in the pre-treatment group and post-treatment group than in the UVB control group.Conclusion Hydrogen may mitigate UVB-induced oxidative damage to skin fibroblasts.

The United Kingdom is highly experienced in the career development and deployment of health care assistants ( HCA) to cope with shortage of nurses. This paper introduced the HCA entry requirements and career development in UK. The entry requirements include quality requirements, basic training and education requirements, practice certificate training and effect evaluation. Career development includes job promotion and transfer to clinical nurses. In view of the current situation of nursing assistant position in China, this paper made valuable recommendations.

OBJECTIVETo investigate the maturation of individual bones on the hand and wrist in children with central precocious puberty (CPP) and idiopathic short stature (ISS).

METHODSHand and wrist films of 25 children with CPP, 29 children with ISS and 21 normal controls were evaluated by conventional Greulich-Pyle (GP) atlas method and individual bone assessment method, in which all twenty bones of the hand and wrist were evaluated based on GP atlas, including 2 radius and ulna, 7 carpal bones, 11 metacarpal and phalangeal bones, the average bone age (BA) was calculated. The differences in groups were analyzed by independent samples t test. The differences between the two methods were analyzed by paired sample t test. The differences between BA and chronological age (CA) were analyzed by ROC with SPSS 17.0.

RESULTSCompared with the normal control group, the advance of BA in the CPP group was 0.70-2.26 y (1.48 ±0.78) by the GP atlas method, while that was 0.28-2.00 y(1.14 ±0.86) by the individual bone evaluation method. In all twenty bones, the advance of metacarpal and phalangeal BA was the greatest [0.34-2.06 y(1.2±0.86)]. In the ISS group,the delay of BA was 0.47-2.91 y(-1.69±1.22) by the GP atlas method, while that was 0.48-2.50 y (-1.49±1.01) by individual bone evaluation method.The delay of carpal BA was the greatest [0.59-2.73 y(-1.66±1.07)] in all twenty bones. In the ISS group and the normal control group, there were no statistic differences between the two methods. In the CPP group, statistic difference was found between two methods. There were no statistic differences for the areas under ROC curves between two methods.

CONCLUSIONThe advance of metacarpal and phalangeal BA is the greatest in CPP group and the delay of carpal BA is the greatest in ISS group.Both methods provide diagnostic information for bone age in CPP and ISS children.

Age Determination by Skeleton ; methods ; Carpal Bones ; diagnostic imaging ; Case-Control Studies ; Child ; Dwarfism ; diagnostic imaging ; Female ; Hand ; diagnostic imaging ; Humans ; Male ; Puberty, Precocious ; diagnostic imaging

AIM: To study the characterization of thyrotropin receptor (TSHR) and its active fragment TSHR aa352-366 as immunogens in BALB/c mice. METHODS: BALB/c mice were injected peritoneally with TSHR aa352-366-KLH (hemocyanin from keyhole limpets) and the mixture of TSHR aa352-366-KLH and guinea pig TSHR every 15 days, respectively. The levels of thyroid hormones and TSHR antibodies and TSHR mRNA were measured, and the pathological changes of thyroid tissue were observed. RESULTS: In the group injected with TSHR aa352-366-KLH, the serum levels of TT_3 and TT_4 decreased (P

Objective: To investigate the current status of job burnout in clinical nurses in a grade A tertiary hospitalin Shaoxing,China and related influencing factors. Methods: In October 2016, the Nursing Burnout Scale (NBS)was used for the investigation of 304 clinical nurses in a grade A tertiary hospital.The contents of the investigation included general data(including age,education background,working years,marital status, frequency of night shifts,professional title, and way of employment), characteristics of working environment,burnout, personality characteristics,coping strategy,and psychosomatic symptoms.SPSS 18.0 was used to conduct Pearson correlation analysis of the scores of each dimension of NBS. A multivariate regression analysis was performed with the demographic features of clinical nurses as the independent variable and the scores of each dimension of NBS as the dependent variable. Results: Among the clinical nurses in this grade A tertiary hospital, the incidence rate of severe burnout was 74%.The Pearson correlation analysis showed that burnout,pessimistic personality,negative coping,and psychosomatic symptoms were positively correlated with working environment(r=0.530,0.316,0.116,and 0.502); pessimistic personality and psychosomatic symptoms were positively correlated with burnout(r=0.618 and 0.675); psychosomatic symptoms were positively correlated withpessimistic personality(r=0.540); negative coping was negatively correlated with pessimistic personality(r=-0.145).The multivariate linear regression analysis showed that department(Department of Internal Medicine or Department of Surgery,B=-0.364 and -0.428)and frequency of night shifts(<6 times/month and 6-10 times/month,B=0.199 and 0.256)were influencing factors for the score of working environment; department(Department of Internal Medicine or Department of Surgery, B=-0.350 and -0.360)was an influencing factor for the score of burnout; 1-3 working years(B=-0.238)was an influencing factor for the score of pessimistic personality; married state,1-3 working years,and department (Department of Internal Medicine or Department of Surgery)were influencing factors for the score of psychosomatic symptoms(B=0.263,-0.301,-0.322,and -0.391). Conclusion There is a high incidence rate of job burnout among clinical nurses in this grade A tertiary hospital,which is associated with burnout,working environment, pessimistic personality,and psychosomatic symptoms.Marital status,working years,department,and frequency of night shifts are major influencing factors for job burnout.