Purpose To establish a new method for detecting p16INK4a in cervical tissues with time-resolved fluoroimmunoassay (TRFIA).Methods 126 cases of paraffin imbedding tissues of cervix were selected for immunohistochemistry (IHC) of EnVision two-step and TRFIA.Results There were 20 cases of no intraepithelial lesion or malignancy,24 cases of low-grade squamous intraepithelial lesion (LSIL),53 cases of high-grade squamous intraepithelial lesion (HSIL) and 29 cases of squamous cell carcinoma (SCC).In the groups of no intraepithelial lesion or malignancy,LSIL,HSIL and SCC,p161NK4a positive was seen in 1,19,53 and 28,respectively.TRFIA test results displayed p16INK4a positive in 3,17,50 and 27 cases,respectively.Positive of p16 using by TRFIA in no intraepithelial lesion or malignancy,LSIL,above HSIL was 15.00％,70.83％ and 93.90％,respectively (P ＜ 0.01).Conclusion TRFIA is suitable for detecting of p16INK4a protein and demand low detection equipment,p16INK4a expression detected by TRFIA may helpful for large scale detection in various clinical institution.

Objective:To observe influence of mitochondrial manganese superoxide dismutase (MnSOD ) on vascular alkaline phosphatase (ALP) activity ,transcription factor :Runx2 and Osterix expression levels in MnSOD‐Tg mice model of vascular calcification .Methods :A total of 20 wild type (WT) mice were selected as WT group and 20 Mn‐SOD‐Tg mice were enrolled as MnSOD‐Tg group ,then mice model of vascular calcification was established .Vascu‐lar ALP activity ,expression levels of transcription factor Runx2 and Osterix were compared and analyzed between two groups .Results:After successful model establishment ,compared with WT group ,there were significant reduc‐tions in ALP activity [ (75.89 ± 4.17) vs .(61.32 ± 3.12)] , expression levels of transcription factor Runx2 [ (0.928 ± 0.016) vs .(0.694 ± 0.007)] and Osterix [ (0.472 ± 0.036) vs .(0.257 ± 0.013)] in MnSOD‐Tg group , P<0.01 all .Conclusion:Manganese superoxide dismutase may be improving atherosclerosis via reducing activity of vascular alkaline phosphatase ,expression levels of transcription factor Runx2 and Osterix .

OBJECTIVE:To improve the quality of ADR reports. METHODS:By using intensive hospital monitoring mode, ADR reports before and after the application of the system would be collected and factors as report quantity,type,time and quality of reports were analyzed statistically. RESULTS:It provided rapid reporting function after the implementotion of the system. Total quantity of reported ADR cases increased from 589 to 748,and the proportion of all the serious ADR reports increased from 62.8%to 11.76%. The score of repert quality increased from 93.64 to 98.36,the proportion of time-out reports increased from 94.05% to 97.33%,with statistical significance (P<0.05). CONCLUSIONS:Rapid reporting system of ADR in the hospital is beneficial to improve the efficiency and guarantee the quality of the reports. It also can expand the coverage of the monitoring network,and can lay the foundation for drug safety scientific evaluation and monitoring.

AIM:To observe the expression of TLR2 and TLR4 on mast cells (MCs) in the periapical tissues from different types of human chronic periapical diseases , and to analyze the role of TLR 2 and TLR4 on tryptase-positive MCs in the immunopathogenesis of human chronic periapical diseases .METHODS: A total of 60 donors, including healthy control group , periapical granuloma group and periapical cyst group , were enrolled in the study .The periapical tis-sue specimens were fixed in 10%buffered formalin and stained with hematoxylin and eosin for histopathology , or stained with double-immunofluorescence for identification of TLR 2-tryptase and TLR4-tryptase double-positive MCs in the periapical tissues.RESULTS:Compared with the healthy control , the densities of TLR2-tryptase and TLR4-tryptase double-positive MCs in periapical tissues were significantly increased in human chronic periapical diseases (P<0.01).The densities of TLR2-tryptase and TLR4-tryptase double-positive MCs in periapical cyst group were significantly higher than those in peria-pical granuloma group (P<0.01).CONCLUSION:TLR2 and TLR4 were expressed on the MCs in the periapical tissues of human chronic periapical diseases .TLR2-tryptase and TLR4-tryptase double-positive MCs may participate in the patho-genesis of chronic periapical diseases .

A novel flow-injection chemiluminescence (FI-CL) method free of CL reagent was developed for the determination of captopril based on its enhancing effect on the CL derived from diperiodatoargentate(III)-sulfuric acid system. Compared with the conventional CL system, the CL system based on trivalent silver was characterized of good selectivity for the absence of CL reagent. The CL mechanism was discussed through CL spectra and UV–vis absorption spectra. The conditions of the FI-CL system were investigated and optimized. Under the optimal conditions, the relative CL intensity was linear with the captopril concentration in the range of 0.3–15.0 μg/mL. The detection limit for captopril was 0.05 μg/mL, and the relative standard deviation (n=11) was 2.0% for 5.0 μg/mL captopril. The proposed method was applied to the analysis of captopril in tablet and human urine with the recoveries of 83.1%–112.5%, and the relative standard deviations of 0.5%–4.4%. The results obtained by the proposed method agreed well with those obtained from HPLC method. The proposed method is fast, convenient, and cost-effective for the determination of captopril in medicine and biological samples.

Aim To investigate the effect of genistein on apoptosis in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods MTT assay was used to detect the inhibition rate on breast cancer MDA-MB-231 cells of genistein.Hoechst 33258 staining was applied to determine the effect of genistein on morphology of MDA-MB-231 cells.qRT-PCR was employed to detect the mRNA expression of EGFR in MDA-MB-231 cells.Western blot was utilized to determine the expression of Bcl-2, Bax, caspase-3, EGFR, Akt, and p-Akt.The expressions of Akt and p-Akt proteins in breast cancer MDA-MB-231 cells were detected after treated with Akt activator insulin, genistein and in combination with insulin.Results Genistein inhibited the viability of breast cancer MDA-MB-231 cells in a time-dependent manner.The results of Hoechst 33258 staining showed a typical apoptotic morphological changes of MDA-MB-231 cells after treatment of genistein for 36 h.qRT-PCR showed that the mRNA expression of EGFR in MDA-MB-231 cells decreased after treated with genistein for 36 h.The expression levels of Bcl-2, EGFR, Akt, p-Akt, ERK, p-ERK were significantly down-regulated(P<0.01) compared with control.While, the expression of Bax, caspase-3 was significantly up-regulated (P<0.01).It was observed that p-Akt was significantly activated after the treatment of Akt activator insulin (P<0.01), however, significantly down-regulated (P<0.01) when treated with genistein.Conclusion Genistein could inhibit the growth of triple-negative breast cancer MDA-MB-231 cells and induce apoptosis, which probably involves regulating EGFR/PI3K/Akt signaling pathway.

OBJECTIVE To explore the outcomes of CO2 laser assisted stapedotomy with artificial stapes prostheses in the treatment of advanced otosclerosis. METHODS Between January 2010 and January 2014, 15 patients (16 ears) diagnosed as advanced otosclerosis accepted CO2 laser assisted stapedotomy with artificial stapes implantation in our department. The averaged preoperative air conduction threshold of the speech frequency was 70.21 dB HL, the averaged bone conduction threshold was 38.49 dB HL, the averaged air-bone gap (ABG) was 31.72 dB HL. All cases were followed up for more than 6 months after operation. RESULTS All cases accepted auditory follow up after 6 months postoperatively. The speech frequency average air conduction threshold was 43.7 dB HL, the average bone conduction threshold was 28.95 dB HL, the average ABG was 14.75 dB HL. The ABG≤20 dB was achieved in 9 ears (56.3%) and ABG closure (≤10 dB) was achieved in 6 ears (37.5%). No cases appeared intractable vertigo, sensorineural hearing loss, secondary facial paralysis and other serious complications. CONCLUSION CO2 laser assisted technique reduced the probability of serious complications of stapedotomy, most patients with hearing level improved significantly. It's a safe, practical, relatively economical choice for advanced otosclerosis.

ObjectiveTo investigate the efficiency of autologous transplantation of peripheral blood stem cells for treatment of patients with diabetic lower limb ischemia.MethodsEighteen patients of type 2 diabetes with diabetic lower limb ischemia (30 legs) were treated by autologous transplantation of peripheral blood stem cells.ResultsThe limb pain, cool feeling and numbness feeling improved significantly after PBSC transplantation,the improvement rate were 96.7％, 100.0％ and 95.8％ respectively.Intermittent claudication was also relieved significantly, total remission rate was 76.9％.The ABI and TcPO2 of patients increased significantly at 3 months after transplantation.After the transplantation ABI raised from 0.60 ± 0.11 to 0.71 ±0.12(t =-6.882, P ＜ 0.01) .93.3％ of patients' TcPO2 raised in different degrees.The foot infections were well controlled.Ulcer or toes gangrene got better or healed.No obvious complications or adverse reaction were observed after the transplantation.ConclusionAutologous transplantation of peripheral blood stem cells shows to be a simple, safe and effective method in treating patients with diabetic lower limb ischemia.

Objective To establish intellectual critique of nursing thesis course in postgraduate nurse,and to evaluate teaching effectiveness and the reasonableness of the course.Methods A total of 44 postgraduate nurses were involved in the study,which were carried out through action research of planning,acting and observing,and reflecting.After the course,researchers evaluated the reasonableness of the course and teaching effectiveness through the self-control.Results There was significant difference in the scientific research ability score of research object before and after the course:(60.09±16.26) scores vs.(71.27±13.22) scores,t=-3.264,P＜0.01.But there was no significant difference in the critical thinking ability scores of research object before and after the course:(295.55±29.71) scores vs.(292.71±26.67) scores,t=0.478,P＞0.05.Conclusions The course of intellectual critique of nursing thesis can improve the research ability of postgraduate nurse,but the role of the critical thinking ability remains to be discussed.And the course needs to be further improved.

Objective To investigate the temporal and spatial distribution of Schistosoma infection of population and its risk factors in Eastern Dongting Lake area in 2012 and 2014,so as to provide the reference for formulating effective intervention mea-sures. Methods Junshan District was selected as a study field in Eastern Dongting Lake area. The method of spatial autocorre-lation analysis was applied to analyze the change of spatial distribution of Schistosoma infection in Junshan District in 2012 and 2014. The spatial regression model was fitted to detect the risk factors for human infection. Results The livestock infection rate in 2013 was lower than that in 2011. The average infection rate of schistosome was reduced to 0.55%in 2014. The spatial auto-correlation existed on the distribution of schistosomiasis in Junshan District in both 2012 and 2014 and 4 high incidence villages were identified. The results of the spatial error model showed that the prevalence of human infection was positively correlated with the infection rate of the livestock and the area of the susceptible environment in 2012. The spatial lag model showed that the prevalence of human schistosomiasis was positively correlated with the area of the susceptible environment ,but not with the in-fection rate of livestock. Conclusion The measures involving grazing prohibition and phasing out cattle and sheep are remark-ably effective and should continue on the basis of the current spatial distribution of schistosomiasis in this area.