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.

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.

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.

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 analyze the Envelope (E) gene of type 1,2,3 dengue virus isolated fromGuangzhouin2010, andtoinvestigatetheinfectionsourceandvirusgenotypes.MethodsEighty-five serum samples were collected from 85 patients in acute phase of dengue fever.Dengue virus was cultured and isolated by C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced.The phylogenetic tree was drawn by neighbor-joining method.The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiologic data.ResultsSix strains of type 1 dengue virus,two strains of type 2 dengue virus and six strains of type 3 dengue virus were isolated from 85 samples.The E gene sequence of these strains was obtained by sequencing.The phylogenetic analysis showed that type 1 and 3 dengue virus belonged to two genotypes (Asian and South Pacific ocean,India subcontinent and Southeast Asia/South Pacific ocean,respectively),and type 2 dengue virus belonged to one genotype (Malaysia/India subcontinent).ConclusionIt's presumed that all strains of type 2 dengue virus are imported,four strains of type 1 dengue virus are imported and four strains of type 3 dengue virus arc imported,the remaining two stains of type 1 and two stains of type 3 dengue virus need mosquito intermediary research further to prove their origins.

Objective To evaluate the feasibility of perfusion weighted imaging (PWI) in the differentiation of recurrent glioma and radiation-induced brain injuries. Methods Fifteen patients with previously resected and irradiated glioma, presenting newly developed abnormal enhancement, were included in the study. The final diagnosis was determined either histologically or clinicoradiologically. PWI was obtained with a gradient echo echo-planar-imaging (GRE-EPI) sequence. The normalized rCBV ratio[CBV(abnormal enhancement)/CBV(contralateral tissue)], rCBF ratio[CBF(abnormal enhancement)/CBF(contralateral tissue)]and rMTT ratio[(MTT abnormal enhancement)/MTT(contralateral tissue)]were calculated, respectively. The regions of interest (ROIs) consisting of 20-40 mm2 were placed in the abnormal enhanced areas on postcontrast T1-weighted images. Ten to fifteen ROIs measurements were performed in each lesion and the mean value was obtained. Mann-Whitney test was used to determine whether there was a difference in the rCBV/rCBF/MTT ratios between glioma recurrence and radiated injuries. Results Nine of the 15 patients were proved recurrent glioma,6 were proved radiation-induced brain injuries. The mean rCBV ratio[2.87(0.70-4.91)]in glioma recurrence was markedly higher than that[0.70(0.12-1.62)]in radiation injuries (Z=-2.55,P＜0.05). The mean rCBF ratio[1.89(0.64-3.96)]in glioma recurrence was markedly higher than that[0.56(0.12-2.08)]in radiation injuries (Z=-2.08,P＜0.05). The areas under rCBV and rCBF ROC curve were 0.893 and 0.821. If the rCBV ratio ≤0.77, the diagnosis sensitivity of radiation-induced brain injuries was 100.0%;If ≥2.44, the diagnosis specificity of recurrent glioma was 100.0%. Conclusion PWI was an effective technique in distinguishing glioma recurrence from radiation injuries and rCBV and rCBF ratios were of great value in the differentiation.

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.