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.

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.

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 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.

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 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.

Objective To investigate the number and activation status of circulating mucosal associated invariant T (MAIT) cells in patients with sepsis.Methods Flow cytometric method was used to determine the number and percentage of MAIT cells and their expression of activation molecule CD69.Results The circulating MAIT cells dropped significantly at early stage of sepsis in septic patients.The MAIT cells in the sepsis group continued to express high levels of CD69 on day 1 and day 3 after admission to ICU.Conclusions Remarkable decrease of circulating MAIT cells may portend the possibility of sepsis in patients with severe infection.

Objective To study the application value of MSCT in preoperative evaluation of rectal cancer before surgery.Methods Clinical materials of 146 patients with colonoscopy and clinical proved rectal cancer were recruited.MSCT were performed with plain and triphasic dynamic contrast enhancement before surgery.The value of MSCT was evaluated by comparing the results of pre-operative evaluation with the surgical results.Results 130 cases accepted surgical treatment:3 cases of transanal local excision,28 cases of Miles operation,3 cases of Hartmann operation,96 cases of Dixon.Preoperative evaluation of MSCT:33 cases of Miles,4 cases of Hartmann,88 cases of Dixon,5 cases of local excision of the anus.The accuracy on MSCT preoperative evaluation was 84.62%. MSCT preoperative evaluation consistency was consistent with postoperative outcome(k=0.653,χ2=225.352,P=0.000).Conclu-sion It is high accuracy for MSCT in preoperative evaluation of rectal cancer,and it can provide important significance for preopera-tive selection of surgical procedures.

Biological mass spectrometry has been developed for the largE-scale protein identification. The successful identification of protein in proteomic study is based on an effective match of MS data to the sequence in database. Because of the diversity and heterogeneity of protein modification, the experimental data obtained by mass spectrometry does not match the theoretical value sometimes, which makes about 90 percent or more of the tandem mass spectra not be effectively identified. This has become one of the most important technique issues to be resolved in current proteome research. The N-terminal cyclization of peptides, as one of a variety of modification introduced in sample preparation, has been preliminarily studied in this work. The result showed that N-terminal cyclization occurred at the most of the glutamine(Q) or carbamoylmethyl-cysteine(CAM_C) residues and the reaction is often incomplete or partial, both types of peptides could often exist in its respective state at the same time, and the behavior of modified peptides in revered phase chromatography is also changed. The success rate of protein identification could be obviously improved if adding the N-terminal cyclization modification in the database searching. These results will be very helpful in the mass spectrometric data analysis of proteomic study.