Enterovirus 71 (EV71) is the main causative agent of hand, foot, and mouth disease (HFMD). This article presented the inhibitory activity of pentapeptides on the EV71 infection in rhabdomyosarcoma (RD) and suckling mice. The EV71 VP1 capsid protein expression levels and mRNA levels were analyzed by Western blotting and real-time PCR. The antiviral activity of pentapeptides in vivo was evaluated by weight changes and EV71 VP1 protein expression levels in intestines of suckling mice. Results revealed that the pentapeptide P010157 was able to inhibit EV71 replication in RD cells. After being incubated with the P010157 at a concentration of 100 microg x mL(-1) for 48 h, the level of EV71 vp1 mRNA in RD cells decreased by (92.0 +/- 6.3)%. The estimated EC50 was 2.2 microg x mL(-1). P010157 was able to inhibit EV 71-induced cytopathic effect (CPE) in RD cells. The cytotoxic activity of the compound was evaluated against RD cells by MTS assay. The results showed that P010157 had no obvious toxicity. In addition, the treated mice with P010157 did not exhibit weight loss, as was observed in untreated mice. EV71 replication reduced significantly as revealed by Western blotting. These findings suggest that P010157 could prevent EV71 proliferation in vitro and in vivo. P010157 is a novel compound for antiviral therapies against EV71, which merited further investigation.
Animals ; Antiviral Agents ; pharmacology ; Capsid Proteins ; genetics ; metabolism ; Enterovirus A, Human ; physiology ; Enterovirus Infections ; metabolism ; Humans ; Mice ; Oligopeptides ; pharmacology ; RNA, Messenger ; metabolism ; Rhabdomyosarcoma ; metabolism ; pathology ; virology ; Tumor Cells, Cultured ; Virus Replication ; drug effects

Enterovirus 71 (EV71) is the main causative agent of hand, foot, and mouth disease (HFMD). This article presented the inhibitory activity of pentapeptides on the EV71 infection in rhabdomyosarcoma (RD) and suckling mice. The EV71 VP1 capsid protein expression levels and mRNA levels were analyzed by Western blotting and real-time PCR. The antiviral activity of pentapeptides in vivo was evaluated by weight changes and EV71 VP1 protein expression levels in intestines of suckling mice. Results revealed that the pentapeptide P010157 was able to inhibit EV71 replication in RD cells. After being incubated with the P010157 at a concentration of 100 microg x mL(-1) for 48 h, the level of EV71 vp1 mRNA in RD cells decreased by (92.0 +/- 6.3)%. The estimated EC50 was 2.2 microg x mL(-1). P010157 was able to inhibit EV 71-induced cytopathic effect (CPE) in RD cells. The cytotoxic activity of the compound was evaluated against RD cells by MTS assay. The results showed that P010157 had no obvious toxicity. In addition, the treated mice with P010157 did not exhibit weight loss, as was observed in untreated mice. EV71 replication reduced significantly as revealed by Western blotting. These findings suggest that P010157 could prevent EV71 proliferation in vitro and in vivo. P010157 is a novel compound for antiviral therapies against EV71, which merited further investigation.

Objective To investigate the effect of elemene on the radiosensitivity of A549 cells and its possible molecular mechanism.Methods The effect of radiosensitivity was detected by colony forming assay.The protein expressions of DNA-PKcs,Bcl-2 and P53 were detected with Western blot.The correlation between the protein expression of DNA-PKcs and Bcl-2,DNA-PKcs and P53 was analyzed.Results Elemene had radiosensitizing effect on A549 cells,with the SERDo and SERDq 1.54 ± 0.20 and 1.43±0.15,respectively for 10 μg/ml elemene,and 1.63 ±0.32 and 1.75 ±0.19,respectively for 20 μg/ml elemene.Compared with irradiation group,the expression of DNA-PKcs was reduced significantly in 10,20 μg/ml elemene combined with radiation group ( t =7.52,8.33,P ＜ 0.05 ),so was for Bcl-2(t =10.74,11.33,P ＜0.05).The expression of P53 protein increased significantly (t =-9.25,7.66,P ＜0.05).There was a remarkable negative correlation between the expression of DNA-PKcs and P53(r =-0.569,P ＜0.05),and a remarkable positive correlation between DNA-PKcs and Bcl-2 (r =0.755,P ＜0.05 ).Conclusions Elemene has radiosensitizing effect on A549 cells,which might be related to down-regulation of DNA-PKcs gene expression,up-regulation of P53 and down-regulation of Bcl-2.

Objective To screen radiosensitizing-related genes mediated by elemene in lung adenocarcinoma A549 cells by using gene chip. Methods MTT test was used to calculate the IC50 of elemene. ①The effect of radiosensitivity was detected by colony forming assay. A549 cells were divided into 2 groups: radiation group and radiation + elemene group. Oligonucleotide chip was used to screen the gene expression changes of A549 cells from these 2 groups. The up-regulated gene Egr-1 and the down-regulated gene CyclinDl were selected to undergo RT-PCR so as to confirm the reliability of the result. Results MTT test showed the elemene inhibited the proliferation of the A549 cells dose-dependently. The IC50 value of elemene on the A549 cells was 120 mg/L. ②10 mg/L elemene had radiosensiting effect on A549 cells.The values of SERDO and SERDq obtained from the survival curve were (1.54±0. 20) and (1.43±0. 15 )respectively. Gene chip screened 122 differentially-expressed genes, including 89 up-regulated genes and33 down-regulated genes. ③These altered genes could be related to cell structure, substance metabolism,cell proliferation, cell differentiation, signal transduction, material transport, DNA repair, apoptosis,immune response and so forth. The RT-PCR results of Egr-1 and Cyclin D1 were consistent with the genenchip analysis.Conclusions The mechanism of elemene enhancing the radiosensitivity of lung adenoearcinoma A549 cells is the result of participation and collaboration of multiple genes. Further study of the newly-discovered differentially-expressed gene helps find out new radiosensitizational targets of elemene.

Objective: To study the effect of β-elemene combined with radiotherapy on the expression of KU70 mRNA and apoptosis of lung adenocarcinoma cell line A549. Methods: A549 cells were divided into the control group (c), irridation group (IR), β-elemene group (0.1×IC50 and 0.2×IC50 I.e. 10 μg/mL and 20pg/mL β-elemene) and β-elemene combined with irridation group (0.1×IC50 + IR and 0.2×IC50 + IR). IC50 was obtained through MTT method and cell survival rate was analyzed by colony formation test. Cell cycle and apoptosis were detected with flow cytometry. The expression level of KU70 mRNA was detected by RT-PCR. Results: MTT assay showed that IC50 value of A549 cells was 120 μg/mL. The number of cell clones and survival rate of β-elemene groups were declined significantly. The radiosensitivity of A549 cells can be enhanced by β-elemene. The flow cytometry confirmed that the ratio of G_2/M and apoptosis were significantly in-creased under the effect of β-elemene, statistically different from those in the control group (P <0.05). The expression level of KU70 mRNA in β-elemene with radiotherapy group was declined significantly, statistically different from those in the con-trol group (P<0.05). Cendusion: The rediosensitization effect of β-elemene on A549 cells is associated with induction of cell apoptosis and down-regulation of KU70 mRNA expression.

Objective To study the influence of Dahuang granule on the level of motilin in plasma and gastrin in serum of patients with severe acute pancreatitis (SAP). Methods Sixty-four patients with SAP were divided into Dahuang granule group (32 cases) and control group (32 cases), treated with Dahuang granule and routine therapy respectively. The level of motilin in plasma and gastrin in serum of the two groups were detected by radioimmunoassay method. Results After treated for one week, the level of motilin in Dahuang granule group was significantly higher than that in the control group (P

Objective To study the value of echocardiography in the diagnosis of myocardial infarction(MI) and its complica tions.Methods The examination data in 140 cases of MI were collected from January 2012 to January 2016 and performed the analysis and comparison.Results In the ultrasound examination results in 140 cases,120 cases appeared varying degrees of segmental ventricular wall motion abnormalities,its diagnostic rate to MI was 85.71％,which of EKG was 78.57％,the MI detection rate of echocardiography and ECG combined examination was 90.71％,which was higher than that of single use of echocardiography or ECG.With the coronary angiogra-phy results as the control,the detection rate of echocardiography for the coronary arterial lesion sits was 82.86 ％.Among 140 cases of MI,22 cases of complications(15.71 ％) were de-tected out by echocardiography,including 7 cases of true ventricular aneurysm(5.00 ％),4 cases of left ventricular mural thrombosis(2.86 ％),1 case of ventricular septal perforation(0.71％) and 10 cases of ischemic mitral regurgitation(7.14％).Conclusion Echocardiography has higher accuracy in the MI diagnosis,com-bining with ECG can further improve the diagnostic rate.

OBJECTIVE:To establish a method for the determination of 9 residual organic solvents in blonanserin as methanol, alcohol,isopropyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene. METHODS:Headspace gas chromatography was adopted. The determination was performed on DB-624 capillary column using temperature pro-gramming. The inlet temperature was 150 ℃,and flame ionization detector was used with temperature of 250 ℃. High purity nitro-gen was used as carrier gas with flow rate of 2.8 mL/min. The split ratio was 1:1,and headspace sample size was 1 mL. Head-space heating temperature was 90 ℃,and equilibration time was 35 min. RESULTS:The linear ranges of methanol,alcohol,iso-propyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene were 6-1500 μg/mL(r=0.9998),10-2500 μg/mL(r=0.9999),10-2500 μg/mL(r=0.9998),0.82-205 μg/mL(r=0.9994),1.2-300 μg/mL(r=0.9995), 0.58-145 μg/mL(r=0.9994),10-2500 μg/mL(r=0.9999),1.44-360 μg/mL(r=0.9996),1.78-445 μg/mL(r=0.9995),respec-tively. The limits of quantitation were 17.71,6.02,3.17,7.45,1.53,0.69,0.93,1.01,0.22 μg/mL;the limits of detection were 5.89,1.90,1.05,2.48,0.51,0.23,0.31,0.33,0.07 μg/mL,respectively. RSDs of precision and stability tests were all lower than 3.0%,and isopropanol was found in repeatability test (RSD=2.1%). The average recoveries ranged 96.67%-102.66%(RSD=1.9%,n=9),96.00%-101.83%(RSD=1.9%,n=9),97.17%-101.50%(RSD=1.4%,n=9),96.97%-102.44%(RSD=2.2%,n=9),95.83%-103.33%(RSD=2.5%,n=9),95.83%-100.28%(RSD=1.9%,n=9),98.17%-101.25%(RSD=1.0%,n=9),96.55%-102.30%(RSD=1.9%,n=9),96.30%-102.22%(RSD=1.8%,n=9),respectively. CONCLUSIONS:The method is simple,rapid,accurate and suitable for simultaneous determination of 9 residual organic solvents in blonanserin as methanol,alco-hol,isopropyl alcohol,acetonitrile,dichloromethane,hexane,ethyl acetate,tetrahydrofuran and methylbenzene.

Purpose To explore the relationship between the mutations of epidermal growth factor receptor ( EGFR) and KRAS genes and clinicopathological characteristics in patients with non-small cell lung cancers (NSCLC). Methods Clinical samples from 431 NSCLC patients were obtained for EGFR and KRAS gene analysis. PCR based direct DNA sequencing was used to investigate mutations in exon 18-21 of EGFR gene and codon 12 and 13 of exon 2 of KRAS gene. Results The overall EGFR mutation rate of primary NSCLC was 53. 6% (231/431) in this study cohort and eight cases showed double EGFR mutations. Mutation rates in female and male were 65. 2% (122/187) and 46. 9% (98/209), respectively. The mutation rate was higher in patients with non-smokers and adeno-carcinoma and adenosquamous carcinoma subtypes than in their counterparts (P<0. 05), with the percentage of 57. 2% (124/216), 60. 3% (199/330), 42. 9% (6/14), respectively. In squamous cell carcinomas and other subtypes, EGFR mutation rates were 11. 6% (5/43) and 11. 1% (1/9), respectively. The EGFR mutation types included exon 18 point mutations (4. 0%, 9/227), exon 19 deletion mutations (4. 5%, 101/227), exon 20 insert or point mutations (9. 7%, 22/227) and exon 21 point mutations (41. 4%, 94/227). Activating mutations of KRAS gene were detected in 7. 8%(31/396) of NSCLC. Twenty-eight patients showed codon 12 mutations ( G>T, G>A, G>C) , and three patients had codon 13 mutations ( G>A, G>T) . Most of these mutations were G to T transversion (64. 5%, 20/31). Conclusion Polymerase chain reaction-direct sequencing is a reliable and effective method for the detection of the EGFR and KRAS gene mutation in NSCLC patients. The mutation rate of EGFR is higher in Chinese patients, especial-ly in non-smoking female patients with adenocarcinoma.

Objective Oral and Maxillofacial trauma Diagnosis and Treatment Expert system (OMDTES)was used in maxillofacial trauma PBL teaching to improve the quality of PBL teaching,and perfeet the assessment criteria, Methods OMDTES was applied in two procedures of maxillofacial injury PBL teaching activity:the preparation of lesson plan and assessment criteria.Then a questionnaire was designed to assess the effect of this new mode of PBL teaching.Results The new teaching model is welcomed by students and can improve students'interest in learning,the satisfaction of teaching and ameliorate method of assessment of PBL teaching.Conclusion Applying OMDTES in oral and maxinofacial injury PBL teaching has special advantages in raising the leavnevs'activity of learning,training their ability of chnical thinking and analysis.And it is worthy of further research and amelioration.