Objective To explore the role of Bcl-2 and Caspase-3 in modulating apoptosis of ER-negative breast cancer cells induced by tamoxifen. Methods ER-negative breast cancer cell lines MDA-MB-231 were treated with 10.0?M tamoxifen for 12, 24, 36,48, 60 hours. The rate of cell apoptosis with or without caspase-3 inhibitor Ac-DEVD-CHO, and protein expression of Bcl-2,Bax were determined by flow cytometry, and the activity of Caspase-3 was examined with fluorophotometry. Results The expression of Bcl-2 was down-regulated, the activity of Caspase-3 and the rate of cell apoptosis were increased by TAM time-dependently, and the rate of apoptosis reached its peak at 48 hours. The expression of Bcl-2 was (negatively) correlated with activity of caspase-3. Tamoxifen, however, did not affect Bax protein expression. Ac-DEVD-CHO, a caspase-3 inhibitor, blocked the activation of caspase-3 and inhibited cell apoptosis (induced) by tamoxifen. Conclusions TAM could induce apoptosis in ER-negative breast cancer cells via (mitochondria) pathway by down-regulating Bcl-2 expression, and the activation of Caspase-3 might play an (important) role in the process of tamoxifen-induced apoptosis of ER-negative breast cancer cells.

OBJECTIVE To report the investigation on validating clinical moist sterilizer by applying wireless thermo-loggers in accordance with the advanced Euro and British standards.METHODS The validation had been implemented with microbiological tests and thermometric tests to measure the temperature distribution of the chamber of sterilizer,the temperature and time relation inside the tested package of small and challenge loads,the pressure of the chamber and the temperature beside the drainage.RESULTS The measurements presented the relation between temperature and time measurements of the spots inside the chamber of the tested sterilization loads.CONCLUSIONS The measurement results indicate directly the operation condition and the temperature-time relation inside sterilization loads.These measurements are helpful for controlling sterilization quality.The results of microbiological tests are negative,which are consistent with that of thermomeric tests.

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

Objective To explore the utilization and effectiveness of ultrasound screening for neural tube defects (NTDs),so that to provide scientific evidences for the secondary prevention of NTDs.MethodsFour hundred and fifty-nine women who delivered or gestated NTDs babies or fetuses were randomly selected from Shandong Province and Shanxi Province,and the related information was collected with structured questionnaire by trained interviewers.Results Of the 459 cases,the ultrasonography utilization rate was 98.7%,and 6 cases (1.3%) never took examinations by ultrasonography during the whole pregnancy period.The total diagnosis rate of ultrasound screening for NTDs was 85.9%,and those of anencephalus,spina bifida and encephalocele were 96.4%,79.6% and 73.8% respectively (P<0.05).The average diagnosis week of NTDs was 24.0 and those of anencephalus,spina bifida and encephalocele were 21.2,27.1 and 24.7 respectively (P<0.05).The detection rates of NTDs before 16 weeks,16-20 weeks,20-24 weeks,24-28 weeks and after 28 weeks were 14.1%,49.4%,46.3%,49.2% and 52.1% respectively (P<0.05).The detection rates of NTDs in hospitals,maternal and child care service centers and family planning centers were 46.4%,52.0% and 28.1% respectively (P<0.05).The detection rate of NTDs by two-dimensional ultrasound equipment was 41.3% and 83.3% by three-dimensional ultrasound equipment (P>0.05).Conclusions The detection rates of NTDs and the subtypes by ultrasonography are low at different pregnant periods and in different medical institutions.It is important to increase the utilization rate of ultrasound screening by pregnant women and improve the NTDs diagnostic level of primary health care institutions,so that to improve the efficacy of secondary prevention strategy for NTDs in China.

Objective:To evaluate the clinical performance of CAD/CAM zirconia all-ceramic restorations.Methods:287 all-ce-ramic Zirconia restorations in 206 patients were included in a 3-year prospectively survey with California Dental Association Standard (CDA)as a reference.The effects and the related factors such as restoration type,tooth region,fiber reinforced composite application on survival rate were analyzed.Kaplan-Meier survival analysis and Log-Rank test were used for data analysis.Results:Chipping frac-tures in 5 restorations,intense gingivitis at 4 restorations and periapical inflammation for 3 restorations were observed during the obser-vation period.The 3-year cumulative survival rate (CSR)of CAD/CAMzirconia restorations was 95.7%,The differences among the CSR of single crowns(96.3%),linked crowns (93.6%)and fixed partial dentures(95.7%)were not statistically significant(P >0.05).The difference was not statistically significant(P >0.05)between the CSR of the anterior region group(94.5%)and posterior region group(96.3%)as well as difference(P >0.05)between the CSR of fiber reinforced composite group (95.0%)and without fiber reinforced composite group(96.1%).Conclusion:The cumulative survival rate of CAD/CAM zirconia all-ceramic restorations is high.Chipping fracture is the main reason of failure.Restoration type,tooth region and fiber reinforced composite have no significant effect on the survival of zirconia restorations.

Objective To investigate the incidence of electrocoagulation syndrome after endoscopic submucosal dissection (ESD) in the colorectal laterally spreading tumors (LST) and the risk factors. Methods Data of 51 patients with coloretral LST,treated with ESD from January 2010 to May 2014 at Shengjing hospital affiliated to China Medical University,were reviewed.The incidence of electrocoagulation syndrome was analyzed and logistic regression was used to evaluate risk.Results The incidence of electro-coagulation syndrome was 9.8%(5 /51).The incidence of tumors in the rectal area(7.1%,2 /28)was lower than that of the left half colon (12.5%,1 /8),and the right colon (13.3%,2 /15).Multivariable logistic regression analysis showed that the independent risk factors for the development of electrocoagulation syndrome were LST located in non-rectum area (OR =1.655,P <1.655),lesion size larger than 25 mm (OR =1.028, P <0.05),the operation time longer than 129 min (OR =1.016,P <0.05),age older than 62 year old (OR =0.987,P <0.05).Conclusion For the patients aged over 62 year old,lesion size larger than 25 mm,the operation time longer than 129 min and LST located outside the rectum,the mucous membrane should be separated from the muscularis propria in the ESD procedure to reduce electrocoagulation time as much as possible. In the postoperative period,patients need fasting,fluid replacement support,and prevention of post endoscopic submucosal dissection electrocoagulation syndrome (PEECS).

Objective The aim of this study was to investigate the expression of MAGEA4 and EB1 proteins in lung cancer tissues and their correlation with clinicopathological features and prognosis. Methods A total of 136 patients with lung cancer in our hospital were enrolled. The expression levels of MAGEA4 and EB1 at levels of mRNA and protein were measured by real-time fluo-rescence reverse transcription and immunohistochemistry. The correlation between MAGEA4 and EB1 expression and clinical patholog-ical features,and prognosis were analyzed by χ2 test and Cox regression analysis. Results The expression of MAGEA4 and EB1 mR-NA in lung cancer tissues was higher than those in adjacent tissues(P<0. 05). The positive rates of MAGEA4 and EB1 in lung canc-er tissues were higher than those in adjacent tissues(P<0. 05). The expression of MAGEA4 and EB1 proteins in lung cancer tissues was higher than those in adjacent tissues(P<0. 05). The positive rates of MAGEA4 and EB1 proteins were not significantly correlated with age(P>0. 05),but they were related to the maximum diameter,pathological grade,TNM stage,infiltration depth,lymphatic vas-cular infiltration,lymph node metastasis and recurrence(P<0. 05). The 3-year survival rate and total survival time of MAGEA4 and EB1 negative group were significantly higher than those of MAGEA4 and EB1 positive group(P<0. 05). Lymphatic vascular infiltra-tion,lymph node metastasis,MAGEA4 positive and EB positive were independent risk factors for prognosis of patients with lung cancer (P<0. 05). Conclusion The positive expression rates of MAGEA4 and EB1 proteins in lung cancer tissues are increased,and their high expression may be related to the occurrence and development of lung cancer. Lung cancer patients with negative expression of MAGEA4 and EB1 proteins can obtain better prognosis.

Objective To understand the situation of college students' knowledge, attitude and practice (KAP) of innovation and entrepreneurship activities and analyze their influencing factors, in order to provide a scientific basis for further promotion of innovation and entrepreneurship activities. Methods A self-administered questionnaire survey using cluster sampling method was conducted among 850 undergraduates in a medical university in May, 2018. The demographic characteristics and questions pertaining to the KAP of innovation and entrepreneurship activities were collected, The weighted scores of KAP were calculated using the Delphi method. Chi-square test, multivariate logistic regression and linear regression analysis were used to analyze the data. Results A total of 850 questionnaires were sent out and 800 valid questionnaires were collected. The average scores and pass rates for the knowledge, attitude and practice of the 800 college students were (0.54 ±0.14), 38.1％; (0.56 ±0.23), 51.5％; and (0.37 ±0.19), 12.6％, respectively. Knowledge: the pass rate for the concepts related to innovation and entrepreneurship was 70.5％, and the pass rate for the policies related to innovation and entrepreneurship was 17.2％. The pass rate of senior students and excellent students were higher than those of junior students and students of lower academic competence (P=0.000, P=0.004). Attitudes: the pass rate for attitude towards the relationship between innovation and entrepreneurship and personal development was 63.6％, the pass rate for attitudes towards participation in innovation and entrepreneurship was 49.8％, and the pass rate for attitudes towards the school continuing to carry out innovation and entrepreneurship related activities was 23.8％. Practice:the pass rates for participation in scientific research and academic activities and social practice , voluntary cultivation of innovation ability and self-employment were 8.5％, 62.9％, 11.9％and 27.3％, respectively. The pass rate of senior grades was higher than that of junior grades (P=0.001). Scores for knowledge and those of attitude are positively correlated. Scores for practice and those of knowledge and attitude are positively correlated. Conclusion The higher pass rates for medical college students' attitude toward innovation and entrepreneurship activities as compared to those for knowledge and practice indicates high eagerness toward participation in innovation and entrepreneurship activities; a high rate knowledge is conducive to the cultivation of attitudes and the improvement of participation in activities . Therefore , it is necessary to strengthen the efforts in carrying out innovation and entrepreneurship activities , improve publicity and education, and promote the awareness-raising of innovation and entrepreneurship.

Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.

Objective: To investigate the role of human papillomavirus (HPV) 16 early genes E2 and E6 and heterogeneous nuclear ribonucleoprotein (hnRNP) E2 and their interaction effects in the progression of the cervical cancer. Methods: Women with normal cervix (NC), low cervical intraepithelial neoplasia (CIN Ⅰ) and high cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ) from the cervical lesions cohort in Jiexiu County of Shanxi Province from June 2014 to September 2014, and patients with cervical squamous cell carcinoma (SCC) treated at the Second Hospital of Shanxi Medical University in the same period were enrolled in this study. There were 257 participants, about 67 NC cases (26.07%), 69 CIN Ⅰ cases (26.85%), 68 CIN Ⅱ/Ⅲ cases (26.46%), and 53 SCC cases (20.62%), respectively. The information of demographic characteristics, life health habits and cervical lesions were collected by using the structured questionnaire. Cervical exfoliated cells and cervical biopsy tissues were collected to detect the infection of HPV16 and the protein expression levels of hnRNP E2, HPV16 E2 and E6. According to the median-value of the protein expression levels of hnRNP E2, HPV16 E2 and E6 and E2/E6 ratio in the NC group, the study participants were divided into the high and low expression groups/ratio groups. The multivariate logistic regression model was used to analyze the correlation between HPV16 early gene E2 and E6, hnRNP E2 and cervical cancer. The interaction effect was analyzed by using the generalized multifactor dimensionality reduction (GMDR) model. Results: The ages of NC, CIN Ⅰ, CIN Ⅱ/Ⅲ and SCC groups were (47.00±9.07), (47.64±7.35), (46.37±8.67) and (51.26±8.03) years old, respectively. The multivariate logistic regression model analysis showed that the HPV16 E2 low expression, E6 high expression and E2/E6 low ratio could increase the risk of CIN Ⅱ/Ⅲ, about OR (95%CI) values 11.11 (1.63-75.56), 8.00 (1.28-50.04), and 9.75 (1.22-77.72), respectively and SCC, about OR (95%CI) values 14.22 (2.11-95.88), 10.33 (1.67-64.00), and 12.38 (1.56-97.91), respectively. The hnRNP E2 low expression could increase the risk of CIN Ⅱ/Ⅲ and SCC, about OR (95%CI) values 3.35 (1.39-8.10) and 5.53 (1.54-19.88). The result of GMDR showed that there were interaction effects of the hnRNP E2 low expression, HPV16 E2 low expression and HPV16 E6 high expression in both CIN Ⅱ/Ⅲ and SCC groups. Conclusion The HPV16 E2 low expression, HPV16 E6 high expression and hnRNP E2 low expression could increase the risk of high-grade cervical intraepithelial neoplasia and cervical cancer, and they might have an important interaction effect in the progression of the cervical cancer.