Objective To evaluate the clinical value of cardiovascular dual-phase scan of 256-slice spiral CT in diagnosis of left atrial appendage (LAA) thrombus before radiofrequency ablation in patients with atrial fibrillation. Methods A prospective study was conducted. Thirty-six patients with atrial fibrillation being prepared to undergo radiofrequency ablation admitted to the Fifth Central Hospital of Tianjin from October 2015 toJuly 2016 were enrolled, they were scanned using dual-phase cardiovascular protocol of 256-slice spiral CT, and then trans-esophageal echocardiography (TEE) was performed for the definite diagnose of thrombus. In the first phase of cardiac CT, the intelligent tracking method was used to determine the delayed time; in the second phase cardiac CT scan, 85 seconds was confirmed as the delayed time; TEE as the golden standard was used to evaluate the value of dual-phase CT in definite diagnosis of LAA thrombus.Results LAA low density filling defect was discovered in 5 patients in the first phase CT scan, the CT scan in the second phase, the filling defect still existed, and the diagnosis of LAA thrombus in 3 patients was made (of them 2 cases after TEE examination were diagnosed definitely as LAA thrombus, and the echo in 1 case was smoke-like on TEE, being at pre-thrombus status), 2 cases were confirmed as pseudo-filling defects (afterwards, their diagnosis was confirmed as pre-thrombus status because the echo shown on TEE was smoke-like). Two patients were confirmed as true thrombi on TEE, and there were 3 patients diagnosed as pre-thrombus state by TEE because of their echo smoke-like. TEE was used as the golden standard for diagnosis of thrombus, the following indexes could be calculated: in the first phase, the sensitivity of using CT scan to diagnose LAA thrombus was 100.0%, the specificity 91.2%, positive predictive value (PPV) 40.0%, and negative predictive value (NPV) 100.0%; while in the second phase of using CT scan for diagnosis of LAA thrombus, the above indexes were 100.0%, 97.1%, 66.7%, 100.0% respectively; the CT Kappa coefficient of the second-phase was larger than that of the first-phase CT (0.898 vs. 0.739), the difference being statistically significant (P < 0.05).Conclusions Dual-phase cardiovascular protocol of CT can detecte of LAA thrombus/pre-thrombus state, the PPV is significantly elevated after the second phase of CT scan for diagnosis of thrombus, and the consistency between the second phase CT diagnosis of thrombus and TEE diagnosis is higher than that between the first phase CT and TEE, therefore, using dual-phase cardiovascular protocol of 256-slice spiral CT in diagnosis of LAA thrombus in patients with atrial fibrillation before radiofrequency ablation has relatively high application value.

Objective To design and screen small interefere RNA (siRNA) targeting of HOXA7,and to investigate the effect of the siRNA on human lung cancer LETP-a-2 cells proliferation and apoptosis in vitro.Methods Three pairs of siRNA targeting of HOXA7 and one pair of siRNA for negative control were transfected respectively into LETP-a-2 cells through cationic liposome.The mRNA and proteion expression levels of HOXA7 were observed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The effect of HOXA7 siRNA on growth and apoptosis of LETP-a-2 cells were measured by MTT and flow cytometry.Results All the three pairs of siRNA could inhibit HOXA7 expression effectively,among which siRNA2 got the best effects,the silence rates were (57.344±4.743) ％ on mRNA level and (52.219±0.550) ％ on protein leval.The proliferation was inhibited and the apoptosis was promoted by the siRNA targeting HOXA7 in LETP-a-2 cells,among which siRNA2 got the favourite results,the inhibitory rate was (48.144±4.992) ％ and the apoptosis rate was (26.613±0.612) ％.Conclusion The siRNA2 targeting of HOXA7 enrolls in promoting apoptosis and inhibiting grows of LETP-a-2 cells,indicating that manipulation of HOXA7 expression may be a potential therapeutic strategy for cancer.

Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase-9, Bax mRNA and protein were tested by RT-PCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase-9 and Bax mRNA and protein were significantly higher than those of other two groups (P<0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the down-regulation of BCR-ABL and the up-regulation of caspase-9 and Bax.

Objective To screen siRNAs that can effectively inhibit Apollon gene expression and determine the cellular functions of those siRNAs.Methods A chemical synthesis method was used to synthesize 3 siRNA sequences against different sites of Apollon.They were transfected into the human breast cancer MCF-7 cells by using Lipofectamine 2000.mRNA level of Apollon was determined by reverse transcription-polymerase chain reaction (RT-PCR).Cellular immunity fluorescence quantitative analysis combined with confocal laser technology was used to determine the protein level of Apollon.Methyl thiazolyl tetrazolium bromide (MTT) assay and flow cytometry were used to determine the effects of siRNA targeting Apollon on proliferation and apoptosis of MCF-7 cells,respectively.Results Three pairs of siRNA could significantly inhibit Apollon mRNA expression,at the inhibition rates of (36.201±11.629) ％,(67.308±7.686) ％and (47.123±12.000) ％,respectively (P ＜ 0.05).After tranfection by siRNA2,Apollon protein fluorescence intensity was (14.97±2.08) ％ compared with control cells.The cell proliferation MCF-7 was inhibited by (73.361±2.118) ％and apoptosis was increased by (28.793±0.743) ％.Conclusions Screened siRNA2 effectively silences Apollon gene expression,effectively inhibits the proliferation and increases the apoptosis of MCF-7 cells.This provids the foundation for its clinical application in cancer therapy.

Background and purpose:MicroRNA-486-5p (miR-486-5p) has been demonstrated to play an important role in many kinds of tumor, however, there are few reports about the relationship between miRNA-486-5p in gastric carcinoma. This study was aimed to explore the effect of miR-486-5p on the proliferation, apoptosis and migration abilities of the human gastric cancer cell line SGC7901.Methods:Quantitative real-time PCR (qRT-PCR) analysis was performed to detect the expression of miR-486-5p in the SGC7901 and GES-1 cells, miR-486-5p over-expressing plasmid was constructed and transfected into the human gastric carcinoma cell line SGC7901 using LipofectamineTM2000. The expression of miR-486-5p of the transfected cells was measured by qRT-PCR, the proliferation level of SGC7901 cells was detected by MTT method, the apoptosis rate of the cells was measured by lfow cytometry and the in vitro migration abilities of SGC7901 cells by transwell test. Results:The miR-486-5p expression in SGC7901 cells was down-regulated compared with GES-1 cells. The expression of miR-486-5p in SGC7901 cells that was transfected miR-486-5p over-expressing plasmid was obviously up-regulated. The proliferation and migration abilities of SGC7901 cells were inhibited signiifcantly, and the apoptosis rate of the cells increased. Conclusion:miR-486-5p can effectively suppress the proliferation and in vitro migration abilities of SGC7901 cells, indicating that miR-486-5p might be used as a target for molecular therapy of gastric cancer.

Purpose:To investigate the adaptation of ovarian cancer cell line SKOV 3ipl cells to hypoxia and its correlation with the expressions of VEGF and Bcl 2 protein.Methods:The monolayer SKOV 3ipl cells plated in culture bottles were placed in an anaerobic system consisting of 85%N 2, 5%CO 2, 10%H 2 with very low oxygen concentration (

Objective To investigate the expression of macrophage migration inhibitory factor (MIF) in serum and renal tissue of septic rats with actue kidney injury (AKI), and to explore the effect of Chinese traditional medicine-Xuebijing injection on MIF expression as well as on acute kidney injury in rats with sepsis. Methods Sepsis model was reproduced in rats with cecal ligation and puncture (CLP).Eighty healthy SD rats were randomly divided into three groups:sham operation group(n=16), CLP model group (n=32), and xuebijing group(n=32). All rats were sacrificed at either 2, or 8, or 24 and or 48 hours after operations.MIF mRNA levels in renal tissues of septic rats were semi-quantified by Real-time PCR.The content of MIF in serum was determined by enzyme linked immunosorbent assay (ELISA). Serum creatinine (Cr) contents were measured by automatic biochemistry analyze. Results Compared with sham operation group, transcription of MIF mRNA in renal tissues of model group were significantly enhanced at 8, 24 and 48 hours after operations (P<0.01). Both contents of MIF and creatinine level in serum of model group rose obviously at 24 and 48 hours after operation (P<0.01);Compared with model group, the transcription of MIF mRNA in renal tissues of xuebijing group decrease obviously at 2, 8, 24 and 48 hours (P<0.01) and both contents of MIF and creatinine in serum of xuebijing group drop remarkably at 24 and 48 hours (P<0.01). Conclusion MIF is a kind of late cytokine which might participate in the pathogenesis of AKI in rats with sepsis.Xuebijing injection can inhibit MIF expression, and possess the protective effects on the kidney in rats with sep-sis.

Objective ①Observing urinary neutrophil gelatinase-associated lipocalin (uNGAL)'s concentration variation under the intervention of sepsis; ②Evaluatingu NGAL' s diagnostic value for early acute kidney injury (AKI).Method Fifty-six SD (Sprague Dawley) rats were randomly (random number) divided into four groups,including 16 rats in model group (CLG),16 rats in Xuebijing group (XBG),16 rats in Huangqi and Chaihu injection jointly applied group (HCG),and 8 rats in sham operation group (SOG).The septic models in CLG group,HCG group and XBG group were established by cecal ligation and puncture (CLP).Then,the rats in HCG group was treated with intraperitoneal injectionby Huangqi and Chaihu injections; the XBG group was treated with intravenous injection by Xuebijing injection; the SOG group was treated with open surgery without CLP.After the CLP,serial urine and serum samples were obtained at baseline (just prior to operation),6 h,12 h,18 h,24 h,36 h,48 h,and 72 h,and were measured by sCr,uCr,uNa,and uNGAL.The line graph of uNGAL' s concentration variation was plotted,based on the time.Diagnostic characteristics of urinary NGAL in predicting AKI were assessed by calculating the area under the receiver operating characteristic curve (AUC).Results After the CLP,the uNGAL of sepsis model rats increased quickly within 6 hours.The time points of each group model reaching their peak were 6 hours after CLP in CLG groupand 24 hours after CLP in HCG group and XBG group.These groups' uNGAL all decreased quickly after the peak.The cuNGAL of sepsis model rats was increased quickly within 6 hours after CLP,reached its peak at 24 hours after CLP.In CLG group,the line graphs of uNGAL or cuNGAL were almost overlapped.There is little difference in the concentration of uNGAL or cuNGAL at each time point (uNGAL:6h,t=0.691; 12h,t=1.627; 18 h,t=0.511,cuNGAL:6h,t =0.371 ; 12 h,t =0.474; 18 h,t =-1.187.Statistical significance of all above value was P ＞0.05).InXBG group,the line graph of uNGAL and cuNGAL were not overlapped,but difference between uNGAL and cuNGAL concentration at each time point was not significant (uNGAL:6 h,t =1.222 ; 12 h,t =1.178 ; 18h,t=1.272; 24h,t=0.918; 36h,t =0.442.cuNGAL:6 h,t =1.482; 12 h,t =1.314; 18 h,t=1.280; 24 h,t =0.280; 36 h,t =0.467.Statistical significance of all above value was P ＞ 0.05).In HCG group,uNGAL of AKI rats were higher than non-AKI rats at each time points since 6 hours later (6 h,t =2.351,P＜0.05; 12h,t=3.086,P＜0.01; 18h,t=2.535,P＜0.05;24h,t=2.150,P＜0.05;36h,t =2.485,P ＜ 0.05),The average cuNGAL of AKI rats and non-AKI rats have statistical significance at 6h,18 h,and 24 h (6 h,t=3.013.P＜0.01; 18 h,t =4.804,P＜0.01; 24 h,t=2.682,P＜0.05).At 6 h,Uout can increase cuNGAL' s ability of predicting AKI' s occurrence in 24 hours (AUC increased from 0.839 to 0.900,P ＜ 0.05).Conclusions The intervention to the sepsis rats have influence on the secretion volume and secretion sequence of NGAL in rat urine.uNGAL and cuNGAL are good predictor of AKI occurrence in sepsis rats.

Background and purpose:miR-196a2 functions as an oncogene during tumor initiation and pro-gression. The up-regulation promotes tumor cell proliferation, invasion and metastasis. Therefore, it is promising to be an important tumor biomarker. The aim of this study was to investigate whether rs11614913, a gene polymorphic site ofmiR-196a2, is associated with the risk of leukemia.Methods:A case-control analysis was employed. Bone marrow or periph-eral blood was collected from 210 leukemia patients diagnosed from Jan. 2009 to Jul. 2015 in Yantaishan Hospital (case group) as well as 250 healthy people who were physically examined during the same period (control group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) was used to detect the genotype of rs11614913. Application test was used to compare the difference in the frequency of each genotype between case group and control group. The odds ratio (OR) of SNP allelic genes was calculated using logistic regression analysis and 95%CI represented the risk of leukemia for each genotype.Results:The distribution differences in the frequency of T/T, C/C, C/T genotype of miR-196a2 rs11614913 between case group and control group were statistically significant (P<0.05). The risk of leukemia for individuals who carried mutant homozygous C/C was 2.661-fold higher than those carried wild-type homozygous T/T, and the difference was statistically significant (P<0.05).Conclusion:ThemiR-196a2 gene polymorphic site rs11614913 was associated with the risk of leukemia. Mutant homozygous C/C or C allelic gene carrying was probably a risk factor for leukemia.

Objective: To investigate the effects of traditional Chinese medicine Xuebijing injection (血必净注射液) on tissue tumor necrosis factor-?(TNF-?) expression and blood coagulation parameters in septic rats.Methods: Wistar rats were subjected to sepsis induced by cecal ligation and puncture(CLP).Ninety-six healthy rats were randomly divided into four groups: normal group,sham operation group,CLP model group,and Xuebijing-treated group.The two latter groups were given respectively intravenous injection of normal saline or Xuebijing injection with the dose of 4 ml/kg at 0.5,12,24,36,48 and 60 hours after the establishment of CLP model.Eight rats were sacrificed at 2,8,24,48 and 72 hours postCLP in the two latter groups.Prothrombin time(PT),activated partial thromboplastin time (APTT),and fibrinogen(Fbg) levels were determined.Tissue TNF-? protein levels in liver and lung tissues were also measured at various intervals.Results: TNF-? protein levels in liver and lung tissues were significantly increased at 2 hours after establishment of CLP model compared with those of the normal group(both P