BACKGROUND: It has been widely reported that the most types of cancer are probably originated from cancer stem cells (CSCs) which are subpopulations of tumor cells. Recent studies also suggested that lung cancer could arise from CSCs. However, the phenotypic characteristics of CSCs in lung cancer have not been precisely described.OBJECTIVE: To systematically analyze the expression of the most common CSC markers in cell line A549.METHODS: A549 cells were cultured for 1 week under the condition of conventional high glucose. After that, flow cytometry was used to assess the expression of putative stem cell markers, including CD133, CD24, CD44 and ABCG2.Cells were then sorted according to the expression of CD44 and CD24 markers by fluorescence-activated cell sorting (FACS) and characterized using their sphere-forming capacity in serum free medium supplemented with several growth factors.RESULTS AND CONCLUSION: A549 cells expressed the CSC markers CD44 and CD24 at 64.23％ and 58.62％,whereas the expression of both ABCG2 and CD133 was around 0.9％. Double-positive CD44/133 populations were rare.CD44+/CD24+ and CD44+/CD24- subpopulations respectively exhibited 54.64％ and 23.38％ expression. CD44+/CD24+ and CD44+/CD24- subsets were sorted by FACS. Both isolated subpopulations formed spheres in the serum free medium supplemented with basic fibroblast growth factor and epidermal growth factor. However, there was no significant difference in the sphere formation efficiency among CD44+/CD24+ and CD44+/CD24- subsets as well as A549 cells. Our findings suggest that CD44 and CD24 cannot be considered as potential markers for isolating lung CSCs in cell line A549, and further investigation using in vivo assays is required.

Objective To investigate the correlation between methylation status and death-associated protein kinase 1 (DAPK1) in SiHa and Hela cell line of cervical carcinoma and intervention of DNA methyltransferase inhibitor 5-azacytidine on the expression of DAPK1 and the proliferation of the cells. Methods DAPK1 methylation status was analyzed using methylation-specific PCR methods. The expression of mRNA and protein of DAPK1 were analyzed by RT-PCR and SABC methods after the treatment with 5-azacytidine. MTT assay was used to observe the changes of proliferation activity of the cells after 5-azacytidine treatment. Results DAPK1 genes were methylated and did not express in SiHa cells in the cervical carcinoma. Its expression could be restored by 5-azacytidine. MTT assay showed 5-azacytidine could weaken the proliferation of cancerous cells. Conclusion DAPK1 methylation plays an important role in the carcinogenesis of cervical cells and can reexpress after the treatment with 5-azacytidine which also restored its inhibitory function on carcinoma.

Objective To observe the structural characteristic of Marshall ligament and coronary sinus of 40 dog hearts. Methods The Marshall ligament and coronary sinus of 40 dog hearts fixed with 10% formaldehyde were observed in gross anatomy. Results The Marshall ligament begins at the right external side edge of left atrium appendage, via the left inferior pulmonary vein to the inner surface of pericardium. The length and the width of Marshall ligament of dog hearts were (2.61?0.95) cm, (0.18?0.04) cm respectively. The angle between Marshall ligament and the coronary sinus was (116?11), and the shortest length between Marshall ligament and left atrium was (0.58?0.52) cm. The length and the width of coronary sinus was (2.84?0.85) cm, (0.28?0.07) cm respectively. There are lots of cardiac fibers of ring (47.5%), vertical (37.5%) and diagonal (15%) shape on the surface of coronary sinus. Conclusion A lot of muscle fibers attaching to the surface of Marshall ligament and coronary sinus in dog hearts are abnormal atrial channels. Dog is the favorable animal for research of focal atrial fibrillation.

Objective To study the feasibility of dual-source CT with dual-energy imaging based on single contrast enhancement. Methods The clinical data of 60 cases of patients who underwent dual-energy CT (DECT)examination of the heart in our hospital were analyzed.statistically.Results The success rate of 60 cases were 100% for displaying all 600 coronary artery segments at all stages.The total mean score of image quality was 4.68 ±0.57,and the good image rate reached 95%.6 cases of patients who ac-cepted DSA examination were unanimous with DECT findings;In 6 cases of patients who accepted SPECT examination,60 segments were no perfusion abnormalities,but DECT iodine figures of 54 segments were normal,and 6 segments showed perfusion defect. Conclusion DECT with dual-energy imaging based on single contrast enhancement can get excellent coronary and myocardial perfu-sion imaging in the circumstance of appropriate heart rate,which has some potentials for clinical application.

Objective To investigate the diversity of mosquito-borne viruses in Xinjiang , China, and to identify mosquitos-borne viruses of medical importance rapidly .Methods The virus-derived RNAs in mosquitos captured in wild were screened and confirmed by using Illumina deep sequencing approach and reverse transcription PCR , respectively .The alignment analysis was performed by using gene sequences from GenBank.Results One hundred and forty-four Culex Flavivirus ( CxFV, Flavivirus genus, Flaviviridae) specific sequences were identified .The overlapping reads were assembled into 7 uncontinuous viral genomic contigs.The gaps between the contigs were further filled by RT-PCR products, which resulted in reconstruc-tion of viral genomic 5′and 3′terminus (687 nt and 411 nt).Phylogenetic analysis showed that the newly identified CxFV belonged to America/Asian genotype , which specifically clustered into a clade with other CxFV strains from China mainland ,sharing 98.2%-99.5%homologies in nucleotide sequences and 99.5%in amino acids sequences among them .Conclusion Illumina deep sequencing approach was successfully applied to arthropod-borne virus surveillance .The recently emerged Culex Flavivirus was detected for the first time in Xinjiang, China.

Objective To investigate the effect of depression on morning blood pressure surge ( MBPS) in maintenance hemodialysis( MHD) patients with hypertension. Methods 67 MHD patients with hypertension were included in this study.All clinical data were collected,and ambulatory blood pressure mo-nitor was used to monitor patient''''''''s blood pressure in 24 hours.Self-rating depression scale(SDS) was used to survey 67 MHD patients,and multivariable logistic regression was conducted to determine independent risk factors for morning hypertension in MHD patients with hypertension. Results There were 14 depression pa-tients in non-morning hypertension group ( 35. 0％) , 18 depression patients in morning hypertension group (66.7％),and the difference was significant(χ2=6.479,P=0.011).In addition correlation analysis showed that age(r=0.027,P=0.044),24 h systolic pressure(r=0.284,P=0.020),24 h diastolic pressure(r=0.278,P=0.023),fasting blood-glucose (r=0.293,P=0.039),low-density lipoprotein cholesterol (r=0.345,P=0.016),and the standard score of SDS(r=0. 276,P=0.024)were significantly correlated with MBPS .Furthermore,multivariable logistic regression showed that the standard score of SDS(OR=1.101,95％CI=1.018-1.191,P=0.016) ,24 h diastolic pressure ( OR=1.070,95％CI=1.007-1.136,P=0.028) and fast-ing blood-glucose (OR=2.127,95％CI=1.118-4.049,P=0.022)were independent risk factors for morning hypertension in MHD patients with hypertension ( OR=1. 101, 95％CI:1. 018-1. 191, P=0. 016 ) . Conclu-sion Depression,24 h diastolic pressure and fasting blood-glucose can increase the risk of morning blood pressure surge in patients with MHD hypertension.

Objective To investigate the relationship between blood uric acid levels and total burden scores(TBS)of carotid plaques in elderly patients with non-dialysis dependent chronic kidney disease(CKD). Methods Seventy-seven elderly patients with non-dialysis dependent CKD were included.All clinical data were collected,and stepwise multiple linear regression was conducted to determine independent factors for TBS of carotid plaques in elderly patients with non-dialysis dependent CKD. Results Compared with the control group,TBS was significantly higher in the smoking group(P= 0.035),hypertension group(P= 0.014)and diabetic group(P= 0.041).In addition,correlation analysis showed that hemoglobin,low-density lipoprotein cholesterol,uric acid,C-react protein and estimated glomerular filtration rate were significantly correlated with TBS(all P<0.05).Furthermore,stepwise multiple linear regression analysis showed that TBS was markedly correlated with low-density lipoprotein cholesterol(P = 0.007,)and uric acid(P = 0.001). Conclusions Uric acid is an independent factor for TBS of carotid plaques in elderly patients with non-dialysis dependent CKD.

Objective To investigate the effect of silent information regulator 2 homologue 1 (SIRT1) activity on angiogenesis in cerebral ischemia rats and its related mechanisms .Methods One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into four groups by random numbers:control group , the cerebral ischemia-reperfusion injury model group ( model group ) , the SIRT1 agonist group (agonist group) and the SIRT1 inhibitor group (inhibitor group), with 30 rats in each group. A model of transient middle cerebral artery occlusion was performed by the suture method .After reperfusion for 24 h, neurological deficit scores were evaluated . Cerebral infarct area after middle cerebral artery occlusion ( MCAO) in rats was determined by staining with triphenyltetrazolium chloride .SIRT1 deacetylase activity was detected by ELISA in ischemic brain tissue.By immunohistochemistry, we observed CD34 expression and detected microvascular density ( MVD) in ischemic cerebral cortex.Immunoblotting was carried out to evaluate protein levels of SIRT 1, vascular endothelial growth factor (VEGF) and erythropoietin (EPO) in ischemic brain tissue.Results Compared with the control group ((13.828 ±0.828) U/L), SIRT1 deacetylase activity was significantly reduced in ischemic brain in the model group ((7.721 ±0.581) U/L,t=8.650,P<0.01).Compared with the model group , SIRT1 deacetylase activity was significantly increased in ischemic brain in the agonist group ((26.165 ±0.971) U/L,t=-26.123,P<0.01). Compared with the agonist group , SIRT1 deacetylase activity was significantly reduced in ischemic brain in the inhibitor group ((17.094 ±1.012)U/L,t=12.848,P<0.01).Neurological deficit score was 2.667 ± 0.516 in the model group.When SIRT1 was activated in ischemic brain , neurological deficit score was significantly lower (1.333 ±0.516,t=4.822,P<0.01) than that of the model group .When SIRT1 activity was inhibited, the neurological deficit score increased significantly ( 2.500 ±0.548, t=-4.147, P<0.01).The cerebral infarction area was 15.473％ ±3.049％ in the model group.When SIRT1 was activated in ischemic brain , the cerebral infarction area was significantly reduced ( 9.152％±1.803％,t=3.188,P<0.05).Immunohistochemical results showed that the brown cells in the ischemic cortex were CD34 staining positive cells.When SIRT1 was activated in ischemic brain , the MVD count significantly increased in ischemic cerebral cortex ( the agonist group:8.167 ±1.941/high power lens ,the model group:3.167 ±0.753/high power lens,t=-6.864,P<0.01).Immunoblotting demonstrated that the activation of SIRT1 increased the protein expressions of VEGF (the agonist group:0.568 ±0.012,the model group:0.468 ± 0.008,t=-11.034,P<0.01) and EPO (the agonist group:0.646 ±0.010,the model group:0.471 ± 0.013,t=-20.952,P<0.01) in ischemic brain.Conclusions Activation of SIRT1 has neuroprotective effect on cerebral ischemia-reperfusion injury rats.SIRT1 can promote angiogenesis in the early stage of cerebral ischemia reperfusion via directly increasing the expressions of VEGF and EPO in ischemic brain .

Objective: To summarize the key points of high SMAS facelift. Methods: Forty-six patients who had received high SMAS facelift were included in this study. All the patients had subcutaneous dissection by classic facelift incision. The temporal region dissection was performed superficial to middle temporal fascia to avoid temporal branches injury. The zygomatic ligaments were released by sharp and blunt dissection at malar prominence and zygomatic arch. After sub-SMAS dissection, a complete SMAS-platysma flap was raised including temporal, malar, lower face, neck, and one third or half of the midface. Results: Forty patients had postoprative follow-up for more than one year. Twenty patients showed " excellent" results, eighteen patients had " good" results , two patients " common" . The good and excellent rate was 95%. Conclusions The key points of high SMAS facelift to achieve ideal outcome depend on the safe dissection of superficial temporal fascia and the release of zygomatic ligaments.

Objective To investigate the feasibility and effect of visualization teaching method using HC visual laryngoscope for the standardized training of endotracheal intubation in emergency department.Methods Forty standard-trainee doctors were divided into two groups randomly.20 standard-trainee doctors in team N were taught using the conventional teaching method of emergency medicine for the standardized training of endotracheal intubation,and the other 20 standard-trainee doctors in team HC were taught by visualization teaching method using HC visual laryngoscope.After 1 month training,comparisons of the success rate of tracheal intubation in airway management simulator,the time consumed for intubation and the number of attempt on successful intubation cases,and the satisfaction score with the teaching mode scored by standard-trainee doctors between the two groups were done.Results The one-attempt success rate and the success rate of tracheal intubation in airway management simulator by the standard-trainee doctors in group HC were significantly higher than those in group N (40％ vs.10％,85％ vs.55％),while the number of attempt and the time consumed for successful intubation were significantly less than those in group N[(1.65±0.61) vs.(2.27±0.66),(79.00±8.35) s vs.(89.36±12.03) s).The satisfaction score with this teaching mode and teaching effect of standard-trainee doctors in group HC were significantly higher than those in group N (8.10±0.74 vs.6.55±1.05,8.15±0.80 vs.6.85±0.91).Conclusions The visualization teaching method with application of HC visual laryngoscope for standardized training of endotracheal intubation in emergency department could improve the success rate and efficiency of tracheal intubation in airway management simulator for standard-trainee doctors,and this teaching method may be better than the conventional teaching method on teaching mode presenting some feasibility.