Objective To observe the clinical effect of recombinant human growth hormone(rhGH) in the treatment of Turner syndrome(TS).Methods The clinical data of 40 children with TS who were treated with rhGH were retrospectively analyzed.Results After treatment for 6 months,the GV,IGF-1 levels [(9.4 ± 2.7) cm/a,(433.9 ± 164.9)μg/L] were significantly higher than those before treatment [(2.5 ± 0.4)cm/a,(186.7 ± 7.33) μg/L],the differences were statistically significant(t =45.562 5,5.061 0,all P ＜ 0.05).The treatment dose of rhGH,initial treatment age had no significant relationship with GV,the GH peak before treatment had no significant correlation with GV.There was no obvious adverse reactions in the treatment process.Conclusion For children with TS,rhGH can improve the height,improve the therapeutic effect,reduce the burden of family and society.Low dose of rhGH in the treatment of TS can achieve better curative effect,the initial treatment age and growth rate in children had no obvious relationship,the insulin growth factor is an important index during the treatment,and the changes of insulin growth should be closely monitored,and adjusting the dosage of the drug in order to achieve the best therapeutic effect.

Objective To study the effect of growth hormone on serum levels of Ghrelin, leptin(LP), insulin like growth factor-1(IGF-1) and insulin-like growth factor binding protein 3(IGFBP3) in children with idiopathic short stature(ISS).Methods 86 patients of ISS who received therapy from September 2013 to September 2015 were selected.According to random number table,those patients were divided into observation group ( n=43) and the control group (n=43).Control group was treated with routine nutrition therapy, while the observation group was added with recombinant human growth hormone.The Ghrelin, LP, IGF-1, IGFBP3 levels and height pre-and post-treatment were compared.Results After treatment, the level of Ghrelin in observation group [ ( 4.85 ±0.41 ) ng/mL ] was lower than that of control group [ ( 5.63 ±0.62 ) ng/mL ] , the level of LP [ ( 6.53 ± 0.78)mg/L] was higher than that of control group[(5.49 ±0.62)mg/L](all P<0.05); the levels of IGF-1[(387.94 ±47.38)ng/mL] and IGFBP3 [(6.74 ±0.73)μg/mL]in the observation group were higher than that of the control group[(243.54 ±36.82)ng/mL,(4.85 ±0.61)μg/mL](all P<0.05); the height in the observation group[(110.17 ±8.12)cm] was higher than that of control group[(114.58 ±9.40)cm](P<0.05).Conclusion Recombinant human growth hormone is well for ISS, which can effectively improve Ghrelin, LP, promote physical growth IGF-1, IGFBP3 levels, promote growth.

Objective To explore the role of extracellular signal-regulated kinase (ERK)in central nucleus of amygdala (CeA)in the mechanism of fentanyl-induced hyperalgesia (OIH)in rats. Methods Step 1:12 healthy male Sprague-Dawley rats,weighing 60-100 g,were randomly divided into OIH and Control group.The mechanical paw withdrawal threshold (PWT)and the thermal paw withdrawal latency (PWL)were tested at pre-and post-OIH induction.Then the level of p-ERK in the CeA was analyzed by Western blotting.Step 2:After successful induction of OIH and catheterization in CeA,another 30 SD male rats were randomly divided into 5 groups (n = 6 each):Group OIH;Group OIH+U0124;Group OIH+U0126(0.1 5 nmol);Group OIH+U0126(0.45 nmol)and Group OIH+U0126 (1.5 nmol),then 0.3 μl of DMSO,U0124 (1.5 nmol),U0126 (0.1 5 nmol,0.45 nmol,1.5 nmol)was given through the catheter separately.PWT and PWL were tested before cathe-terization,at pre-OIH induction,post-OIH induction and 0.5 h after CeA drug administration.After the last test of pain threshold,the rats were sacrificed and CeA tissues were sampled for analyzing the expression of p-ERK by western blot.Results In step 1 compared with control group,PWT and PWL of OIH group were sharply decreased post-OIH induction (P <0.05),concomitant increase of the expression of p-ERK in CeA in OIH group was also observed.In step 2,both PWT and PWL were sharply decreased post-OIH induction (P <0.05).Intra-CeA U0126 injection,but not U0124, reversed both behavioral hyperalgesia and molecular activation of ERK in CeA in a dose-dependent manner (P <0.05).Conclusion ERK plays a pivotal role in the maintenance of fentanyl-induced hy-peralgesia.Targeting inhibition of ERK activation in CeA can alleviate fentanyl-induced hyperalgesia.

Objective To investigate the role of extracellular signal-regulated kinase1/2 (ERK1/2) in the central amygdala on fentanyl-induced hyperalgesia in rats.Methods Thirty-two male SpragueDawley rats, weighing 60-100 g, were randomly divided into 4 groups (n =8 each) using a random number table: control group (group C), fentanyl-induced hyperalgesia group (group H), U0124 group (group U1) , and U0126 group (group U2).A catheter was implanted in the central amygdale.In group C, normal saline was injected subcutaneously, and 6.5 h later dimethyl sulfoxide (DMSO) was injected via the catheter.In group H, fentanyl was injected subcutaneously to induce hyperalgesia, and 6.5 h later DMSO was injected via the catheter.In group U1, hyperalgesia was induced, and 6.5 h later ERK1 inhibitor U0124 1.5 nmol was injected via the catheter.In group U2, hyperalgesia was induced, and 6.5 h later ERK1/2 inhibitor U0126 1.5 nmol was injected via the catheter.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal threshold (TWT) were measured before fentanyl injection, at 6.5 h after injection, and at 30 min after DMSO or U0124/U0126 administration via the catheter (T0-2).After the last measurement of pain threshold, the rats were sacrificed, and the amygdala tissues were sampled for detection of the expression of phosphorylated ERK1/2 (p-ERK1/2) by Western blot in groups C and H.Results Compared with group C, the MWT and TWT were significantly decreased at T1,2in H and U1 groups, and at T1in group U2 (P＜0.05) , the expression of p-ERK2 was up-regulated (P＜0.05) , and no significant change was found in the expression of p-ERK1 in group H (P＞0.05).Compared with group H,the MWT and TWT were significantly increased at T2 in group U2 (P＜0.05) , and no significant change was found in MWT, TWT in group U1 (P＞0.05).Conclusion ERK2 activation in the central amygdala is involved in the development of fentanyl-induced hyperalgesia in rats.

Objective To explore the factors influencing emergency staff′ compliance with standard precautions in the tertiary hospitals, in order to provide a theoretical basis for improving the standard precautions. Methods Using qualitative research method, eight focus groups were conducted with emergency staff in the tertiary hospitals in China from October to December in 2015. Interview data were imported into Nvivo10.0 qualitative analysis software, using content analysis method for its collation and analysis. Results Factors that affect the standard precautions compliance were grouped into five themes: availability and practicality of protective equipment, organizational climate, safety awareness, obstructive elements to the behavior, and trigger elements to the behavior. Conclusions Standard precautions compliance of emergency medical staff is affected by many factors, management departments should actively build a good professional environment and reinforce safety awareness of the emergency staff.

One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.

Objective To compare the molecular characteristics of hantanvirus strains isolated from 1980 to 2002 in Zhejiang Province by PCR and nucleotide sequencing.Methods Total RNA was extracted from hantanvirus infected Vero-E6 cells. The M segment cDNA of hantaan ZJ4 and ZJ7 strains were obtained by reverse transcription and polymerase chain reaction, subsequently cloned into pUCm-T vector and sequencing.Results Two strains ZJ4 and ZJ7 of hantanvirus isolated from Rottus in Zhejiang province were HTN viruses. HTN virus strain Z10 was isolated in 1980. Comparison ZJ4 and ZJ7 with Z10 strains indicated that there were 88.3% and 92.3% homology at the nucleotide level.Conclusion The HTN virus were very conserved. The homology of HTN virus isolated from Zhejiang Province was high nucleotide level, though they were isolated at more than 20-year intervals.

Objective To study the complete genome sequence of Hantavirus ZT71 strain gene isolated in Zhejiang province and explore its evolution. nethods The total RNA was prepared from ZT71 virus infected cells and the RT-PCR products were cloned into T vector, sequenced and analyzed. Results The L, M and S segments of the strain ZT71 genome were 6530,3651 and 1753 nucleotides in length with a single open reading frame individually encoding 2151,1133 and 429 amino acids. The sequence analysis of nucleotides showed that the homology of L, M and S segments of strain ZT71 between those of other strains of Seoul virus could reach 95.5%-99.7%, 84.1%-99.6% and 88.7%-99.5%, respectively. The analysis of the deduced amino acids showed the similar result. The source of strain ZT71 could be traced from the analysis of the phylogentic trees of nucleotides and amino acids, and it should belong to Seoul type of Hantavirus which was also verified serologically. Conclusion The nucleotide sequences and deduced amino acid sequences of L, M and S segments of strain ZT71 are similar to that of those of Seoul type of Hantavirus. And Hantaan type virus used to be prevalent primarily in Zhejiang province,and it would be an endemic area of mixed type of Hantavirus since the discoveries of the viruses of Soeul type in recent years.