Objective To construct the plasmid pSilencer4.1/HtrA1 and stably transfect human trophoblastic cell line HPT-8.Methods We detected the expression of HtrA1 in cell line HPT-8 with immunohistochemical SABC,constructed the plasmid pSilencer4.1/HtrA1,transfected human trophoblastic cell line HPT-8 with plasmid pSilencer4.1/HtrA1 and negative plasmid pSilencer4.1, and observed the cell expression after transfection. Results About 90% of cellular cytoplasm of HPT-8 was dyed brown while the nucleus was negatively stained. Selective concentration of G418 was 200 μg/mL for HPT-8 to be transfected pSilencer4.1/HtrA1.With RT-PCR and Western blot methods,the expression of HPT-8 transfected plasmid Psilence4.1/HtrA1 was significantly downregulated compared with that of cells with negative plasmid and normal cells (P < 0.05 ).There was no significant difference in absorbance value between blank plasmid group and HPT-8 group (P >0.05).Conclusion We successfully constructed stable trophoblastic cell line HPT-8/ pSilencer4.1-HtrA1 with silenced expression of HtrA1,which lays foundation for further research.

Cultured smooth muscle cells from bovine aortic media were incubated with hyperlipemic serum for 14 days. Lipid peroxides in the cells were higher than controls (P

Objective To assess the quality of randomized controlled trials (RCTs) of delirium prevention of elderly published in China.Methods The literatures from China National Knowledge Infrastructure(CNKI),WANFANG Data and VIP Database for Chinese Technical Periodicals were evaluated according to Cochrane collaboration's tool for assessing risk of bias and Jadad scale.Results A total of 53 RCTs were included,14 (26.4％,14/53) described radom number table used to generate the random allocation sequence,4 (7.5％,4/53) conducted experiments in a blinded manner,7 (13.2％,7/53) did not use intentionto-treat to analyse those who did not complete the study,9 (17.0％,9/53) had high risk of other bias,none described allocation concealment mechanism and blinding to participants and intercention implementers.Based on Jadad scales,the score was 1-4,average score was (2.3±0.8),19 (35.8％,19/53) were high-quality literatures.Conclusions The quality of present published literatures is not high,the further domestic studies should be designed high-quality to better improve clinical practice.

AIM: To observe the effect of microRNA-16 (miR-16) on the megakaryocytic differentiation of K562 cells, and to explore the potential mechanism.METHODS:miR-16 was over-expressed or silenced by transfection with miR-16 mimics or inhibitor in K562 cells.The level of miR-16 was detected by real-time PCR.The expression of CD41, CD42b and CD61, as megakaryocytic differentiation markers, was detected by flow cytometry.The effect of miR-16 on the expression of myeloblastosis oncogene ( MYB) was measured by Western blotting, and flow cytometry was performed to confirm whether the effect of miR-16 on expression of CD41, CD42b and CD61 was mediated by MYB.RESULTS:Transfection with miR-16 mimics dramatically elevated the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells.Transfection with miR-16 inhibitor decreased the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells (P<0.05).The expression of MYB was regulated by miR-16, and MYB silencing reversed the regulation of CD41, CD42b and CD61 induced by miR-16.CONCLUSION:miR-16 regulates the megakaryocytic dif-ferentiation of K562 cells by targeting MYB.

Objective: To explore the mechanism of damaging effect of advanced glycation end products (AGEs) on human umbilical vein endothelial cell (HUVEC) in vitro. Methods: The HUVEC was incubated with exogenous AGEs for 12 h, and ergamine (Hi) and catalase (CAT) were used as control. The activity, monolayer permeability, apoptosis rate, biochemical indexes and morphous changes were detected in HUVEC. Results: The activities of HUVEC were dose-dependently reduced by exogenous AGEs(40 mg/L, 120 mg/L and 160 mg/L), meanwhile the monolayer permeability, the malondialdehyde (MDA) content and apoptosis rate were increased，the activity of superoxide dismutase（SOD）was decreased（P < 0.05, respectively). There was no significant difference in damaging effect between exogenous AGEs and Hi(P ＞ 0.05). The damaging effect of exogenous AGEs was obviously inhibited by CAT in HUVEC. Conclusion: Exogenous AGEs induced the damaging effect on vascular endothelial cells, which may be related to the oxidative stress.

OBJECTIVE：To establish a host specific method for the qual ity evaluation of Taxillus chiueusis by evaluating the quality of T. chiueusis from different hosts sources. METHODS ：HPLC was adopted with CAPCELL PAK C 18 MGⅡ column， mobile phase consisted of methanol- 0.2％ phosphoric acid ，volume flow 1.0 mL/min（gradient elution ），detection wavelength of 254 nm，column temperature of 30 ℃. Totally of 16 batches of T. chiueusis from different hosts sources were collected as sample （4 batches from tea host ，maple host ，willow host and mulberry host respectively ）. HPLC characteristic chromatogram was established with TCM Chromatogram Fingerprint Similarity Evaluation System （2004A edition ），and similarity evaluation was performed. The peak was identified by comparison with the reference substance. IBM SPSS 19.0 software was used for cluster analysis and principal component analysis. RESULTS ：Totally 11 common peaks were demarcated ，peak No. 8，10，11 were identified as rutin ，quercetin and quercitin. The similarity of 16 batches of samples was more than 0.9. The chemical components of T. chiueusis from different hosts sources were basically the same （RSDs of relative time of commom peaks were 0.03％-0.40％）， but the contents of the same component were quite different （RSDs of relative peak area of commom peaks were 27.00％-64.20％）. By cluster analysis ，16 batches of T. chiueusis from different hosts sources were divided into 3 types. The results of principle component analysis showed that quercetin ，quercitin and rutin had significant effect on the quality of T. chiueusis ，which could be used as the quality evaluation index ，and the whole quality of maple host was the best （the highest comprehensive score ）. CONCLUSIONS： HPLC fingerprint combined with chemometrics can be used to evaluate the quality of T. chiueusis from different hosts sources.

Objective To investigate and analyse the status and influential factors of palliative care knowledge of nursing undergraduates,then provide theoretical basis for palliative care education. Methods Using random sampling method, 400 nursing undergraduates were investigated by Palliative Care Quiz for Nursing (PCQN) and palliative care attitude questionnaire. Results 366 nursing undergraduates completed the survey whose scores were 8.363 ± 4.240, and 66.9%(244/366) of them had never heard of palliative care. Choosed right rate of PCQN was 41.13% (150/366). The scores of palliative care philosophy and principles 0.298±0.248, pain and symptom control and psychological scores 0.436 ± 0.235 and social and spiritual support scores 0.499 ± 0.318 had significant difference (F=38.866, P=0.000). The scores of palliative care attitude questionnaire were 36.756 ± 3.183, and the average score of all items were over 3. The palliative care attitude scores had significant difference in different grade (F=2.737, P=0.043). Conclusions Nursing undergraduates hold positive attitudes of palliative care, but the cognition degree were at a low level. To promote the development of palliative care, it should be strengthened publicity, education and training in this area. It is necessary to carry out palliative care education in nursing undergraduate students.

ObjectiveTo investigate the diagnostic value of single balloon enteroscopy (SBE) for obscure gastrointestinal bleeding.MethodsA total of 78 SBE procedures was conducted on 72 patients with obscure gastrointestinal bleeding,with 40 via oral route and 38 via anal route.The procedure time,insertion depth and rate of positive finding were recorded.ResultsFor 40 SBE procedures performed via oral route,the mean procedure time was 60 minutes ( 15-110 minutes),and the mean insertion depth was 195 cm at the distal end of Trentz ligament (30-240 cm).For 38 SBE procedures performed via anus,the mean procedure time was 75 minuets (30-120 minutes),and the mean insertion depth was 160 cm at the proximal end of ileocecal valve (50-200 cm ).The whole diagnostic yield of obscure gastrointestinal bleeding was 62.5％.ConclusionSBE is a safe and useful tool for the diagnosis of obscure gastrointestinal bleeding.

The dynamic changes of immunoreactive dynorphin1-13 (ir-Dyn1-13) contents in 18 burned patients were studied by radioimmunoassay. It was found that ir-Dyn began to increase gradually at the onset of burn and remained at a relatively high levels by the time of preliminary wound healing. Plasma ir-Dyn increased gradually after bum and decreased abruptly before death in one severely burned patient who died of respiratory failure at the third week. The results suggest that Dyn may have a protective effect on the burned patients.

Objective To investigate the risk factors of pulmonary metastasis and prognosis of patients with rectal cancer after anterior resection of rectal cancer.Methods The retrospective case-control study was adopted.The clinical data of 421 patients with rectal cancer who underwent anterior resection at the First Hospital of China Medical University from August 2010 to December 2014 were collected.The patients were followed up by outpatient examination and telephone interview once three months in the first postoperative year,once half a year in the second postoperative year,and then once a year.The follow-up included satuses of pulmonary metastasis and survival of patients after anterior resection of rectal cancer.The end point of follow-up was death of the patients or 31 December,2014.The risk factors of pulmonary metastasis and prognosis in patients after anterior resection of rectal cancer were analyzed.Measurement data with normal distribution were presented as x-± s and measurement data with skewed distribution were presented as M(range).The pulmonary metastasis rate/curve and survival rate/curve were calculated and drawn by the Kaplan-Meier method.The comparisons of pulmonary metastasis rate and survival rate were done using the Log-rank test.The univariate analysis was done using the chi-square test and Log-rank test.The multivariate analysis was done by Logistic regression model and COX regression model.Results Of the 421patients,389 patients were successfully followed up with a median time of 34 months (range,11-53 months) and a follow-up rate of 92.40％ (389/421).Ninety-four,168 and 127 patients were detected in Ⅰ,Ⅱ and Ⅲ stages of TNM stage.There were 29 patients diagnosed with postoperative pulmonary metastasis with the diagnosis time of (21 ± 9) months.The 3-year cumulative incidence of pulmonary metastasis after anterior resection of rectal cancer was 2.2％ in patients of Ⅰ stage,3.0％ in patients of Ⅱ stage and 17.4％ in patients of Ⅲ stage,showing significantly increase trend as the increase of the TNM stage (x2=19.927,P ＜ 0.05).The 29 patients with pulmonary metastasis did not receive chemoradiotherapy including 6 patients receiving pulmonary metastatic nodule recection.Nineteen patients were survived and 10 patients were dead.The survival time of patients diagnosed with pulmonary metastasis was (13 ±9)months and the 3-year cumulative survival rate was 75.7％,whereas the survival time of patients without postoperative pulmonary metastasis was (35 ±9)months and the 3-year cumulative survival rate was 94.3％,showing significant difference between the 2 kinds of patients (x2 =25.219,P ＜ 0.05).The univariate analysis showed that the preoperative carcinoembryonic antigen (CEA) level,degree of tumor differentiation,depth of invasion and lymph node metastasis were risk factors affecting pulmonary metastasis after anterior resection of rectal cancer (x2=4.745,7.250,5.379,18.796,P ＜ 0.05),and the multivariate analysis showed that lymph node metastasis was an independent risk factor affecting postoperative pulmonary metastasis [OR =4.167,95％ confidence interval (CI):1.608-10.801,P ＜ 0.05].The univariate analysis showed that the preoperative CEA level,distribution and number of pulmonary metastatic nodule and lymph node metastasis were risk factors affecting the prognosis of patients with pulmonary metastasis after anterior resection of rectal cancer (x2=13.793,7.246,6.284,4.076,P ＜ 0.05),and the multivariate analysis showed that the preoperative CEA level ＞ 5 μg/L was an independent risk factor affecting the prognosis of patients (HR =13.489,95 ％ CI:1.407-129.297,P ＜ 0.05).Conclusions Pulmonary metastasis after anterior resection of rectal cancer is common.Lymph node metastasis is a high risk factor affecting postoperative pulmonary metastasis,and preoperative CEA level ＞ 5 μg/L is an independent risk factor affecting the prognosis of patients with postoperative pulmonary metastasis.