Objective To study the relationship of HPV pseudo-neutralizing titers detected by two different reporter genes: Zoanthus sp. green fluorescent protein (ZsGreen) and secreted alkaline phosphatase (SEAP) , and the relationship between HPV the pseudovirus-neutralizing antibody titer and the antibody titer determined by ELISA method. Methods The plasmids with expression cassettes of the HPV capsid protein L1 and L2 genes after codon optimization and the plasmid with reporter gene (ZsGreen or SEAP) were co-transfected into 293FT cells. The cell lysate supernatants were collected after 48 h culture, then the pseudovirus was purified through POROS column chromatography from the supernatants. After the titer of pseudovirus bulk were measured, HPV-16 and HPV-18 pseudovirus-neutralization assays were carried out for determining the titer of sera collected from immunized mice with HPV candidate vaccine and Gardasil HPV vaccine. Results In statistical analysis, the two reporter gene systems for the detection of the pseudovirus neutralizing antibody titer are highly relevant to each other (Spearman coefficient; r = 0. 760). And their neutralizing antibody titers bear a high degree of correlation with the antibody titer (Spearman coefficient: r= 0.577 and r =0. 741). Conclusion ZsGreen and SEAP pseudovirus neutralizing antibody titers are highly relevant to each other. The neutralizing antibody and the antibody titer are also relevant. These results reveal some mechanism of HPV vaccines to prevent the virus from invading the host cells, and are absolutely useful in the protection efficiency evaluation of the HPV-16 and HPV-18 candidate vaccines.

Objective To establish a method to quantitatively assess melanosome transfer with incorporation of 14C-thiouracil (TU) into nascent melanin. Methods To characterize whether 14C-TU was exclusively incorporated into melanin-producing cells, the same number of mouse melan-a or SP-1 keratinocytes were labeled with 14C-TU for 12 hours and 48 hours, respectively, followed by the measurement of radioactivity. Mouse melan-a melanocytes were pre-labeled with 1 Ci/mL 14C-TU, and cocultured with mouse SP1 keratinocytes to develop an assay system for melanosome transfer to keratinocytes. Following co-culture, the keratinocytes with transferred radioactivity were separated from melanocytes at different time points via two times of differential trypsinization. Transferred radioactivity in keratinocytes, denoting the amount of melanosome transfer, was measured with liquid scintillation counting. Meanwhile, the effects of forskolin, a PKA activator, and nicotinamide on melanosome transfer were also investigated with this assay system.Results The incorporated radioactivity in melan-a cells was 66- or 80-fold as high as that in SP-1 cells,indicating that 14C-TU would be a suitable tracer for melanosome transfer in co-culture with keratinocytes. A purity of 84.5% was achieved for keratinocytes with transferred radioactivity by twice differential trypsinization.As shown by this assay, there was an approximately 0.67-fold decrease in melanosome transfer with the treatment of 1 g/L nicotinamide and 2.3-fold increase with 20μmol/L forskolin treatment. After coculture with SP1 cells for 8-12 hours, melan-a cells developed well-extending dendrites with detectable melanosome transfer, while no proliferation of melan-a cells induced by forskolin was seen. Conclusion An optimized protocol for selective incorporation of 14C-TU into nascent melanin has been successfully applied to the quantitative measurement of melanosome transfer from melanocytes to keratinocytes induced by forskolin or nicotinamide.

Objective　To explore the therapeutic effect of treatment of uterine leiomyoma with different dosages of mifepristone and the effects of mifepristone on sex hormones.Methods　The patients of uterine leiomyoma were divided into two groups,group A included 45 patients who were mifepristone orally given neoplasma,25mg per day for 3～6 months,group B included 30 patients who took mifepristone 12.5mg per day for 3～6 months.The size of uterus and uterine leicomyoma were detected with type B sonarography and serum sex hormones level were tested with radioimmunoassay before the treatment.Results　Amenorrhea was found during the treatment of two groups and after tree months' the symptoms were improved,the size of uterine leiomyoma in group A was meanly decreased by 52.56% and 61.69% respectively,after six months' treatment;The size of uterine leiomyoma in group B was averagely decreased by 48.61% after three months' treatment and by 52.12% after six months' treatment.There was no significant difference between A and B group after three months' treatment(P＞0.05),But there was significant difference between the two groups after six months' treatment and it dependent on the time of the treatment.Contents of serum estradiol and progestagen of the two groups were droped dwon significantly after treatment(P＜0.01).Conclusions　Mifepristone applied to treat the uterine leiomyoma have definitve therapeutic effect,and less side effects.Therefore,mifepristone provides a new avenue to treat uterine leiomyoma and suggesting its best dosage is 25mg every day.

Objective To explore the effects of wound healing and histological changes by utilizing ultrapulse CO2 fractional laser for the treatment of superficial scar in dermal-epidermal junction (DEJ) in rabbit ears.Methods We adopted traditional iodophor treatment and moist exposed burn ointment treatment repairing the wound and utilizing ultra pulse CO2 fractional laser for the treatment of rabbit ears superficial scar.The superficial scars in rabbit ear were induced with ultra pulse CO2 fractional laser.In 1,2,4,8 weeks tissue samples would be taken.Using electron microscopy we observed DEJ ultrastructure and light microscopy to observe the histological changes of the dermal papilla and PAS staining for epidermis basement membrane.Results The wound healing time of moist exposed burn ointment group was shorter than that of iodophor group (P＜0.05).The number of hemidesmosomes and anchoring fibers in the moist exposed burn ointment group on the 8th week was more than the 1th week,The number of hemidesmosomes and anchoring fibers in the iodophor group on 8th week was more than 1th week (P ＜0.05).The number of hemidesmosomes and anchoring fibers in the iodophor group was less than the moist exposed burn ointment group (P＜0.05).Conclusions U1trapulse CO2 fractional laser can cause ultra structural changes in scarring dermal-epidermal junction area,wet wound healing environment promotes the reconstruction of DEJ district organization,and rebuilding the function of the scar skin has the positive significance.

0 05),But there was significant difference between the two groups after six months' treatment and it dependent on the time of the treatment.Contents of serum estradiol and progestagen of the two groups were droped dwon significantly after treatment(P

In order to reinforce the development and sharing of elaborate educational resources and improve the educational quality, the course team completed the transformation and upgrading of the original fine course for the construction of national high-quality resource sharing class. In the con-struction of courses the organic combination of course content, method, skills, quality was focused on, and the transformation and upgrading of the course achieved further optimization of the excellent resources and full sharing.

Objective To explore the efficacy of multiplex ligation-dependent probe amplification (MLPA)combined with fluorescence in situ hybridization(FISH)and comparative genomie hybridization (CGH)combined with FISH in genetic analysis of chorionic villi specimen(CVS)of spontaneous abortion.Methods CGH+FISH and MLPA+FISH were used for genetic analysis of 29 CVS from spontaneous abortion and 6 normal CVS from selective abortion,in the mean time,those results were compared with conventional eytogenetic karyotyping.Results The report time were 40 hours in MLPA+FISH and 120 houm in CGH+FISH.The mean time of chorionic villi culture was(240±72)hours.The successful rate of specimen analysis were 97%(34/35)in CGH,100%(35/35)in MLPA,100%(35/35)in FISH and 91%(32/35)in conventional cytogenetic karyotyping.Apart from 1 case failed in CGH analysis,the results from MLPA+FISH were almost similar to that from CGH+FISH,however,that 1 specimen failed in CGH were detected successfully by MLPA+FISH.The discrepancy rate were 13%(4/31)in CGH+FISH and 12%(4/32)in MLPA+FISH respectively when compared with conventional cytogenetic analysis.Conclusions MLPA+FISH analysis present shorter detecting time and achieve 100%tale of successful report.This combined method was an important adjuvant approach to conventional cytogenetic karyotyping in CVS from spontaneous abortion.

The herpes simplex virus type 1 (HSV-1) infected-cell protein 27 (ICP27) is an essential,highly conserved protein involved in various steps of HSV-1 gene regulation as well as in the shut-off of host gene expression during infection.It functions primarily at the post-transcriptional level in inhibiting precursor mRNA splicing and in promoting nuclear export of viral transcripts.Recently,many novel functions performed by the HSV-1 ICP27 protein were shown,including leptomycin B resistance,inhibition of the type I interferon signaling,regulation of the viral mRNA translation and determining the composition of HSV-1 virions.

Objective The aim of this study was to investigate the impact of constructing national key clinical specialty on high quality nursing service.Methods Using the Colaizzi 7-step analysis method to analyze the information,which through using the phenomenology research methods by making interviews with 14 clinicians,nurses and nursing administrators.Results The clinical medical staffs experienced the construction of key clinical specialist,to some extent,that would promote the development of nursing discipline,enhancing the environment and equipment in hospital,having improvement on the ability of nursing service and quality simultaneously,and then the cooperation between medical staffs and patients were more closely,while nurses bearing increased work pressure,with obviously job burnout.Conclusions The national key clinical specialist construction would effectively launching nursing quality care in depth and improving patient satisfaction with medical care,while the working pressure of nurses and the imperfection of post performance management restricted the development of nursing discipline.

Objective To investigate the clinical efficacy of laparoscopic radical gastrectomy in elderly patients with advanced gastric cancer.Methods The clinical data of 85 elderly patients with advanced gastric cancer who were admitted to the Ningbo First Hospital from January 2012 to June 2014 were retrospectively analyzed.Laparoscopic radical gastrectomy was performed on 46 patients (LRG group) and open radical gastrectomy on 39 patients (ORG group).All the patients underwent primary tumor resection for gastric cancer + D2 lymph node dissection,and the postoperative recovery plans were done according to enhanced recovery program.The volume of blood loss,number of lymph node dissected,operation time,intraoperative arterial partial pressure of carbon dioxid (PaCO2),time to anal exsufflation,indwelling time of gastric tube,time for out-off-bed activity,time for fluid diet intake,postoperative hemoglobin,duration of hospital stay and occurrence of complications in the 2 groups were analyzed.The follow-up by outpatient examination and telephone interview was carried out on patients up to August 2014.The count data were analyzed by the chi-square test and Fisher exact probability.The measurement data with normal distribution were presented as x ± s and analyzed using the t test.The t' test was used if the data were deficient.Results Surgical procedures in the 2 groups were successfully carried out and no perioperative death occurred.There was no conversion to open surgery in the LRG group.The resection margins in all the patients were negative.The operation time and number of lymph node dissection in the LRG group were (239 ±68)minutes and 27 ± 10,compared with (227 ±50)minutes and 26 ± 10 in the ORG group,with significant differences (t =0.919,0.179,P ＞ O.05).PaCO2 in the LRG group was (41 ± 5) mmHg (1 mmHg =0.133 kPa),which was significantly higher than(36 ± 5) mmHg in the ORG group (t =4.745,P ＜ 0.05).The volume of blood loss was (102 ± 44)mL in the LRG group,which was significantly less than (200 ± 120) mL in the OPG group (t' =-4.807,P ＜ 0.05).The postoperative level of hemoglobin in the LRG was (110 ± 15) g/L,which was significantly higher than (98 ± 27)g/L in the ORG group (t' =2.471,P ＜ 0.05).The time to anal exsufflation,indwelling time of gastric tube,time for out-off-bed activity,time for fluid diet intake,duration of hospital stay in the LRG group were (2.6 ± 0.7) days,(2.1 ± 0.7) days,(1.1 ± 0.3) days,(4.1 ± 0.7) days and (11 ± 4) days,which were significantly different from (4.8 ± 1.5) days,(4.0 ± 1.8) days,(4.5 ± 0.6) days,(5.9 ± 1.8) days and (18 ± 3) days in the OR G group (t' =-8.415,-6.206,-33.831,-5.879,t =9.632,P＜0.05).Eight patients in the LRG group and 15 patients in the ORG group had complications,with the incidence of complications of 17.4％ (8/46) and 38.5％ (15/39),respectively,showing a significant difference (x2 =4.748,P ＜ 0.05).Forty-four patients in the LRG group and 36 patients in the ORG gorup were followed up for 2-25 months,1 patient in the LRG group and 2 patients in the ORG group died and others had full recovery.Conclusions Laparoscopic radical gastrectomy could provide a safe and complete tumor resection for elderly patients with advanced gastric cancer compared with open radical gastrectomy,meanwhile,it can improve postoperative recovery and reduce postoperative complications in elderly patients with advanced gastric cancer.