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.

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 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 .

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.

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.

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 assess the efficacy of trastuzumab combined with neoadjuvant chemotherapy in the treatment of human epidermal growth factor receptor-2 (Her-2)positive breast cancer patients,and to explore its influencing factors of prognosis of breast cancer.Methods:The clinical materials of 112 Her-2 positive breast cancer patients were collected and they were divided into combined treatment group (trastuzumab + neoadj uvant chemotherapy)and single chemotherapy group (without trastuzumab ). SPSS 1 9.0 software was employed to calculate and analyze their clinical characteristics.The survival rate and prognosis were analyzed by Kaplan-Meier method,Log-rank test, and Cox regression. Results:Among the 112 Her-2 positive breast cancer patients, 23 cases were treated with trastuzumab,89 cases were treated without trastuzumab.The disease-free survival (DFS)in combined treatment group and single chemotherapy group had significant difference (P=0.012).And there was no significant difference of overall survival (OS)between two groups (P=0.064).The HR negative group had 18 (32.7%)patients with 5-level of Miller and Payne (MP)classification which was higher than HR positive group (5/46,10.9%)(P=0.009).In univariate analysis,the tumor size,node status at diagnosis and node status after operation were the influencing factors of DFS in the Her-2 positive breast cancer patients;the tumor size,node status at diagnosis,MP classification and node status after operation were the influencing factors of OS in the Her-2 positive breast cancer patients.The result of multivariate analysis indicated that the tumor size, node status at diagnosis,and with or without radiotherapy were the independent influencing factors of DFS and OS in the Her-2 positive breast cancer patients. Conclusion:The breast lesions in the Her-2 positive breast cancer patients with negative HR are relieved after treated with trastuzumab combined with neoadjuvant chemotherapy. The combination of trastuzumab and neoadj uvant chemotherapy can significantly improve the DFS and has a positive influence in the prognosis of Her-2 positive breast cancer patients.

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 analyze the correlation between critical thinking ability and self- efficacy for nursing students in university of traditional Chinese medicine, so as to provide references and evidences for nursing quality teaching. Methods A total of 291 nursing students in university of traditional Chinese medicine were investigated with Critical Thinking Disposition Inventory- Chinese Version (CTDI- CV) and General Self- Efficacy Scale (GSES). Results The total scores of critical thinking ability and self- efficacy were (288.24±32.95) and (25.44±5.74). There were consistence between critical thinking ability and self- efficacy (r=0.348, P<0.01). Regression analysis revealed that self- efficacy and interpersonal relations were predictors of critical thinking ability. Conclusions There were consistence between critical thinking ability and self- efficacy. Self- efficacy and interpersonal relations were predictors of critical thinking ability. It is suggested to promote critical thinking ability by enhancing self- efficacy and interpersonal relations, which contributes to improvement of comprehensive quality.