Objective:To investigate the inhibitory effect and its possible molecular mechanisms of MicroRNA-34a(miR-34a) on the human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice.Methods: The human nasopharyngeal carcinoma CNE-2 cell line was cultured in vitro.miR-34a and Scrambled miRNA recombinant plasmids were successfully established and stably transfected into CNE-2 cells.Fifteen six-week-old male nude mice were divided randomly into three groups:miR-34a group(5 mice) ,Scrambled miRNA group(5 mice) ,Blank control group(5 mice).Different CNE-2 cells were subcuta-neously injected on the back near right lower limb.Tumor volumes were examined every 7 days.Mice were executed on the 35 days,and the eventual average tumor volumes and weights were examined.Total RNA and protein were isolated from tumors,and the expression of miR-34a,CDK6,and Bcl-2 mRNA and protein were determined by qRT-PCR and western blot,respectively.Results: The relative expressions of miR-34a was significantly up-regulated in miR-34a transfected group compared to Scrambled miRNA transfected group (P<0.05).Eight subcutaneous xenograft tumor models were successfully established.The nude mice of miR-34a transfected group appeared a smaller tumor volume compared to the other two groups at the beginning of 21th day( P<0.05).The eventual average tumor volumes in miR-34a group,Scrambled miRNA group and blank control group were(351.37±98.19)mm3,(798.75±91.04)mm3 and
(849.62±101.32) mm3 ,respectively,and the eventual average tumor weights in miR-34a group,Scrambled miRNA group and blank control group were(0.81±0.13)g,(1.47±0.21)g and(1.58±0.37)g,respectively.Both the eventual average tumor volumes and weights in miR-34a group were lower compared to the other two groups(P<0.05).qRT-PCR results revealed that the expression of miR-34a in miR-34a transfected group was significantly higher than in the other two groups,while the mRNA and protein expression of CDK6 and Bcl-2 were lower than the other two groups ( P<0.05 ) .Conclusion: miR-34a may inhibit the growth of human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice by down-regulating CDK6 and Bcl-2.

Objective To determine the content of indirubin in Kangbingan Oral Liquid by UV spectrophotometer methods.Methods The content of indirubin was determined by Heltos Gamma ultravioletvisible spectrophotometer with chloroformic solution of 2～8 ?g/mL confected by standard sample of indigotin and indirubin.The sample of negative blank solution was confected by the prescription without Radix Isatidis.The standard sample and the blank sample were determined separately with the wavelength at 500～700 nm.Results Indirubin had the maximum absorption at wavelength of 540.0 nm and indigotin had the maximum absorption at wavelength of 634.5 nm.The linear correlation was available at the range of 0.20～1.40 ?g/mL,and the correct coefficient was 0.999 4.The average recovery rate was 99.05% and RSD was 0.87%(n=6).Conclusion This method is simple and accurate,and can be used to control the quality of Kangbingan Oral Liquid.

Objective:To study the effect of Qingyi Lidan Granule on the serum levels of high mobility group box 1 (HMGB 1),heat shock protein 70 (HSP70),heat shock protein 27 (HSP27) and interleukin-8 (IL-8) of patients with severe acute pancreatitis.Methods:From August 2015 to July 2016,84 patients with severe acute pancreatitis in our hospital were selected and randomly divided into the observation group and the control group according to random number,42 cases in each group.The control group was given routine treatment,and the observation group was treated by Qingyi Lidan Granule on the basis of control group.The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain,serum levels of HMGB1,HSP70,HSP72 and IL-8 in both groups were observed and compared before and after treatment.Results:After treatment,the total clinical effective rate of observation group was significantly higher than that of the control group[92.86％ (39/42) vs 71.43％ (30/42)] (P＜0.05).The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain in the observation group were significantly shorter than those of the control group (P ＜0.05).Before treatment,no significant difference was found in the serum levels of HMGB 1,HSP70,HSP72,IL-8 between groups (P＞0.05).After treatment,the serum levels ofHMGB1,HSP70,HSP72 and IL-8 in both groups were significantly lower than those before treatment (P＜0.05).The levels ofHMGB1,HSP70,HSP72 and IL-8 in the observation group were lower than those in the control group (P＜0.05).Conclusion:Qingyi Lidan Granule could effectively reduce the levels of serum HMGB 1,H SP70,HSP27 and IL-8 and enhance the clinical curative effect of patients with severe acute pancreatitis.

Problem-based Learning (PBL) is a teaching model originating from the construc-tivism theory, which stressed the meaning construction in problem situation by collaborative learning, conversation and completing knowledge. Based on the times background, development of learning theory, teaching situation, cognitive strategies, ideological connotation about constructivism, PBL's practical significance, its teaching forms, teaching resources, internal motivation, and the teaching evaluation were analyzed deeply, which was beneficial to carrying out PBL normatively in medical education, so as to provide the theoretical support for further exploration and innovation of PBL.

Objective To evaluate the application of flow-through rapid hybridization technique and gene chip (HybriMax) on human papiUomavions (HPV) subtype in Chongqing.Methods Cervical tissue samples were taken under the colpnscope form 473 females who had cervical lesion for pathological analysis.The predictive value of HybriMax in cervical abnormality was compared with pathological results,which were used as golden standard.Resuits 13 different subtypes were found and total HPV positive rate was 63.0% (284/473) Among the 17 different subtypes ,the higher positive rote HPV subtypes were HPV16 (23.7%,112/473),HPV58 (12.7% ,60/473),HPV53(7.4% ,35/473).The HPV infection rates were higher with the worse d cervical lesion(X2=77.06,P＜0.01).Conclusions The most frequent subtypes of HPV infection in Chongqing cervical lesion were HPV 16,58.HybriMax was an effective method to detect HPV subtype in clinical.

Objective To evaluate the effect of non-linear blending function for dual-energy CT on image of pulmonary angiogra-phy.Methods 27 patients underwent dual energy CT pulmonary angiography(CTPA).Data obtained with 100 kVp,140kVp,and non-linear blending were divided into group A,B,and C respectively.CT value of emboli were measured.Corresponding signal to noise ratio(SNR )and contrast to noise ratio(CNR)were calculated.One-way Anova analysis and Friedman test were used to ana-lyze statistical significance among these values.Results On quantitative analysis of 27 patients,there was no statistical difference for CT value of CTPA among the three groups(P >0.05).For the noise,CNR and SNR,there were statistically significant among the three groups(P <0.05).Conclusion The non-linear blending function has certain advantages in improved image signal-to-noise ratio and it can be used in CT pulmonary angiography examination for patients suspected pulmonary embolism clinically .

Objective To evaluate the effects of tumor-associated macrophages on the proliferation,invasion and migration of human cutaneous malignant melanoma cells.Methods Cultured U937 human monocytic cells at logarithmic phase were classified into three groups to be pretreated with phorbol ester for 48 hours followed by 48-hour activation by phorbol ester (M polarization),lipopolysaccharide (LPS) at 25 mg/L (M1 polarization),and interleukin (IL)-4 at 15 μg/L (M2 polarization) respectively.Then,enzyme-linked immunosorbent assay (ELISA) was performed to determine the levels of IL-12p70 and IL-10 in the supernatant of these activated cells.A375 human malignant melanoma cells were divided into four groups to be cultured alone or with M-,M1-and M2-polarized macrophages respectively.After additional culture for different durations (24,48 and 72 hours),methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate the proliferative activity,and Transwell assay to evaluate the invasion and migration activity,of the A375 cells.Results The proliferation of A375 cells was accelerated by coculture with M-and M2-polarized macrophages,but inhibited by that with M1-polarized macrophages,with significant differences among the four groups in the proliferative activity at 48 and 72 hours (all P ＜ 0.05),but not at 24 hours (P ＞ 0.05).Invasion assay showed that the number of A375 cells that migrated through Transwell chambers was significantly larger in M2 and M groups (147.00 ± 7.92 and 113.22 ± 8.15 respectively),but smaller in the M1 group (56.44 ± 7.55),than in the control group (84.11 ± 6.07,all P ＜ 0.05).Similarly,migration assay revealed a significant increase in the number of A375 cells that migrated through Transwell chambers in the M2 and M(p) groups (198.33 ± 8.22 and 156.00 ± 8.83 respectively),but a significant decrease in the M1 group (97.11 ± 6.75) as compared with the control group (123.89 ± 7.01,all P＜ 0.05).Conclusions The proliferation,invasion and migration of A375 cells can be accelerated by IL-4-activated M2-polarized macrophages,but decelerated by LPS-activated M1-polarized macrophages.Phorbol ester tends to induce monocytic cells to differentiate into M2-polarized macrophages.

Objective To evaluate the peripheral dose (PD) from megavoltage cone-beam CT (MVCBCT) imaging for head-and-neck region image-guided radiation therapy,to determine the correlation of PD with monitor unit (MU),and to investigate the impact of imaging field size on the PD.Methods Measurements of PD from MVCBCT were made with a 0.65 cm3 ionization chamber placed in a specially designed phantom at various depths and distances from the field edges.The PD at reference point inside the phantom was measured with the same ionization chamber to investigate the linearity between MU used for MVCBCT and the PD.The homogeneity of PD in the axial plane of the phantom were measured.Results PD from MVCBCT increased with increasing number of MU used for imaging and with increasing the field size.The measured PD in the phantom decreased exponentially as distance from the field edges increased.PD also decreased as the depth from the phantom surface increased.There was a strong linear relationship between PD and MUs used for MVCBCT.The PD was heterogeneous,with higher dose at the anterior than the posterior.Conclusions The PD from MVCBCT depend much on the MVCBCT delivery MU and the scan field size.In clinic,using the smallest number of MU allowable and reducing MVCBCT scanning field size without compromising acquired image quality is an effective method of reducing the PD.

ObjectiveTo explore the relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects (NTDs) induced by all-trans retinoic acid (RA)in Kunming mouse. Methods Fifty pregnant mice were randomly divided into control and RA-treated groups.RA-treated mice were fed with 30mg/kg RA dissolved with peanut oil on embryo 7.75 days, while the mice of control group were administrated with an equal volume of peanut oil on the same time. Then all the embryos were sampled from pregnant mice at the 4th, 18th, 42nd, 66th and 90th hour after treatment. In situ hybridization and immunohistochemical staining technique were used to detect the expression of Dishevelled2 and Vangl2 in embryonic neural tube. Results The two proteins both existed in the epithelial tissue of the mouse embryonic neural tube and displayed different expression modes at various developmental stages.Compared with the control group, the RA treated group showed a significant decrease (P≤0.05) at the 18th and 42nd hour and a significant increase (P≤0.05) at the 66th hour in Dishevelled2 protein after maternal treatment, and no significant difference was found at the 90th hour. Compared with the control group, the Vangl2 mRNA expression in the RA treated group displayed a significant decrease (P≤0.05) at the 4th and 18th hour and a significant increase (P≤0.05) at the 66th hour after RA treatment, and no difference was found at the 42nd hour. Compared with the control group, the expression of Vangl2 protein in the RA treated group decreased (P≤0.05) at the 18th and 42nd hour, and increased (P≤0.05) at the 90th hour after RA treatment, no difference was found at the 66th hour. Conclusion Excessive RA may interfere with the normal embryonic neural tube closure by regulating the expression of Dishevelled2 and Vangl2.

Aim To determine the effects of Qiliqian-gxin injected into lateral ventricle on Cardiac function, angiotensin Ⅱ( Ang Ⅱ) , angiotensin converting en-zyme(ACE), angiotensin type 1 receptor(AT1R) and the sympathetic nervous system in the hypothalamic pa-raventricular nucleus of rats with chronic heart failure. Methods Rat model of heart failure was prepared by acute myocardial infarction that was induced by ligation of the left anterior descending coronary artery. Four weeks after heart failure, Qiliqiangxin and Losartan were continuously administered via a syringe pump in-jector connected to lateral ventricle. After four weeks, echocardiogram was used to evaluate the cardiac func-tion and HE was used to observe myocardial tissue morphology, and enzyme linked immunosorbent assay was used to measure plasma norepinephrine( NE) , ser-um NT-proBNP and Ang Ⅱ in the paraventricular nu-cleus. The expression of ACE and AT1 R at mRNA and protein levels in the paraventricular nucleus was deter-mined by Real-time PCR and Western blot, and the RSNA was measured by PowerLab in anesthetized rats. Results Compared with the sham control, the cardiac function was significantly lower while the AngII, ACE, AT1 R expression in the paraventricular nucleus and RSNA were significantly increased in rats with heart failure. Compared with heart failure control, Qiliqian-gxin and Losartan decreased the RSNA and the AngII, ACE, AT1 R expression in the paraventricular nucleus. Conclusion Giving traditional Chinese medicine to the lateral ventricles can decrease the activation of the RAS system, reduce the renal sympathetic nerve activi-ty and improve cardiac function.