Objective To purify,renature and explore the antiviral effects of recombinant cyanovirin-N(CV-N)on herpes simplex virus type 1(HSV-1).Methods The recombinant CV-N was purified with Ni Sepharose column and renatured by dilution method.The antiviral activities of CV-N were carried out in Vero cells by observing cytopathic effect(CPE)and by using MTT colorimetric assay for vital cell rate.Results SDS-PAGE showed that the purified protein was in the position of 11KDa with only one clear band.The renatured CV-N had little cytotoxic effect on Vero cells,it could not directly inactivate HSV-1 infectivity.CV-N not only interfered in adsorption of HSV-1 to Vero cells but also inhibited HSV-1 biosynthesis in the cells,which were more effective than the positive control Acyclovir.Conclusion CV-N exhibited pronounced antiviral activities agaist HSV-1,further development of CV-N might yield novel candidates of antiviral drugs.

Objective To investigate the status and diagnostic value of aberrant phosphatase and tensin homology deleted on chromosometen(PTEN) gene promoter in tissues, peripheral plasma and BALF of lung cancer patients. Methods We analyzed the methylation of PTEN gene in tissues, peripheral plasma and BALF by methylation specific-PCR. Results The frequency of methylation of promoter of PTEN gene was 26.67%(12/45) in lung cancer tissues, 15.56%(7/45) in peripheral plasma, 22.22%(10/45) in BALF, no methylation product was found in lung tissues without cancer, normal plasma and BALF controls (P

In this study,a standard strain of HSV-1(strain SM44)was used to investigate the antiviral activity of the recombinant Cyanovirin-N(CV-N)against Herpes simplex virus type 1(HSV-1)in vitro and in vivo.Cytopathic effect(CPE)and MTT assays were used to evaluate the effect of CV-N on HSV-1 in Vero cells.The number of copies of HSV-DNA was detected by real-time fluorescence quantitative PCR(FQ-PCR).The results showed that CV-N had a low cytotoxicity on Vero cells with a CC50 of 359.03±0.56 μg/mL,and that it could not directly inactivate HSV-1 infectivity.CV-N not only reduced the CPE of HSV-1 when added before or after viral infection,with a 50% inhibitory concentration(IC50)with 2.26 and 30.16μg/mL respectively,but it also decreased the copies of HSV-1 DNA in infected host cells.The encephalitis model for HSV-1 infection was conducted in Kunming mice,and treated with three dosages of CV-N(0.5,5 & 10 mg/kg)which was administered intraperitoneally at 2h,3d,5d,7d post infection.The duration for the appearance of symptoms of encephalitis and the survival days were recorded and brain tissue samples were obtained for pathological examination(HEstaining).Compared with the untreated control group,in the 5mg/kg CV-N and 10mg/kg CV-N treated groups,the mice suffered light symptoms and the number of survival days were more than 9d and 14d respectively.HE staining also showed that in 5mg/kg CV-N and 10mg/kg CV-N treated groups,the brain cells did not show visible changes,except for a slight inflammation.Our results demonstrated that CV-N has pronounced antiviral activity against HSV-1 both in vitro and in vivo,and it would be a promising new candidate for anti-HSV therapeutics.

Objective To investigate the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the apoptosis of A549/DDP cells and the expression of hMLH1 gene.Methods A549/DDP cells were treated with 5-Aza-CdR at 0.5,5,50 μmol/L.The growth curve of A549/DDP cells was investigated by MTT assay.The methylation status of hMLH1 gene was detected by methylation specific PCR (MSP).The expression of hMLH1 mRNA was evaluated by FQ-PCR.The apoptosis rate of A549/DDP cells was analyzed by flow cytometry.Results A549/DDP cells treated with 5-Aza-CdR showed a slow growth in comparison with the control cells,and the growth rates were decreased with the increasing of 5-Aza-CdR concentration.The apoptosis rate after treatment was higher than that before treatment in A549/DDP cells (P ＜ 0.05),and had a positive correlation with 5-Aza-CdR dose (P ＜ 0.001).hMLH1 mRNA expression level was increased in a 5-Aza-CdR concentration dependent manner (P ＜ 0.05).hMLH1 promoter in A549/DDP cells was methylated and hMLH1 mRNA was negatively expressed before treatment,but the mRNA was positively expressed after treatment with 5-Aza-CdR.Conclusions 5-Aza-2'-CdR can induce apoptosis of A549/DDP cells by inducing demethylation of hMLH1 promoter and thereby enhancing hMLH1 gene expression and its tumor suppressor function.

Objective To investigate the application of left transthoracic small incisions in combination with tubular gastrectomy for radical esophageal cancer surgery in elderly patients.Methods Clinical data of 83 patients with carcinoma in the middle or lower third of the esophagus aged ≥ 70 years who had undergone radical surgery at our hospital from January 2012 to September 2014 were collected.Patients were divided into two groups:the tubular gastrectomy group (n=38) which had undergone radical surgery through left transthoracic small incisions in combination with tubular gastrectomy and the stomach group (n =45) which had been treated with esophagectomy through conventional left posterolateral incisions and esophageal reconstruction with the remnant stomach.Clinical outcomes were compared between the two groups.Results Compared with the stomach group,operative blood loss [(204.3±75.4) ml vs.(258.4±80.2) ml,t=2.720,P＜0.05],chest drainage on the first and second postoperative day [(201.7±82.6) ml vs.(320.5±78.1) ml,(150.8±83.0) ml vs.(244.6 ± 81.2) ml,t =3.221 and 3.189,respectively,each P＜0.05],pain scores on the third postoperative day [(3.73 ± 1.02) vs.(0.24 ± 1.15),t =2.858,P ＜ 0.05],incidence of arrhythmia (13.2％ vs.33.3％,x2 =4.585,P＜0.05),pulmonary complications (18.4％ vs.46.7％,x2=7.353,P＜0.05) and incidences of gastroesophageal reflux and gastric emptying disorders (23.7％ vs.55.6％,0.0％ vs 11.1％,x2 =8.654 and 4.493,both P＜0.05) were less or lower in the tubular gastrectomy group.The two groups had no significant difference in operation time,length of hospital stay,anastomotic leakage,anastomotic stenosis or 1-year survival rate (each P ＞0.05).Conclusions Radical surgery with small incisions through the left chest combined with tubular gastrectomy shows highly favorable clinical outcomes in elderly patients with middle or lower esophageal cancer,and can significantly reduce surgical trauma,decrease postoperative complications,improve the quality of life,and help patients recover after surgery.

Objective To evaluate the effect of perioperative administration of Ambroxol combined with Ipratropium on elderly lung cancer patients undergoing thoracoscopic surgery.Methods 82 lung cancer patients aged ≥70 years who were scheduled for thoracoscopic lung resection were randomly assigned into 2 groups:the observation group(n=42) and the control group(n=40).Patients in the observation group were treated with 90 mg Ambroxol(mucosolvan) by intravenous drip and Ipratropium(atrovent) by atomizing inhalation,while patients in the control group were treated with an equal volume of 0.9％ sodium chloride solution.Pulmonary function and changes of arterial blood gases at admission,before surgery and 5 days after surgery,incidences of postoperative atelectasis and pulmonary infection,the rate of return to the intensive care unit(ICU),and the length of postoperative hospital stay were compared between the 2 groups.Results Compared with the control group,the percent predicted forced expiratory volume in 1 second (FEV1％),the ratio of forced expiratory volume in 1 second to forced vital capacity(FEV1/FVC％),the percent predicted maximum ventilation volume per minute (MVV％),and the arterial oxygen pressure (PaO2) were increased in the observation group before surgery [(87.0±8.2)％ vs.(80.6±7.6) ％,(90.4±6.4)％ vs.(81.4±7.0)％,(87.1±5.6)％ vs.(74.6±6.9) ％,(86.6±6.2) mmHg vs.(81.7±6.9)mmHg,t=2.477,2.588,2.937,3.405,respectively,allP＜0.05].TheFEV1％,FEV1/FVC％,MVV％ and PaO2 were higher in the observation group than in the control group 5 days after treatment [(76.4±9.2) ％ vs.(67.3±10.2) ％,(74.7±9.1) ％ vs.(63.0±11.2) ％,(69.5±9.2)％ vs.(60.1±9.2) ％,(79.5±11.5) mmHg vs.(70.1±11.8) mmHg,t 2.583,2.987,2.778,2.666,respectively,all P＜0.05].The incidences of postoperative atelectasis,pulmonary infection and the length of postoperative hospital stay were lower or less in the observation group than in the controlgroup[7.1％ vs.25.0％,11.90％ vs.32.50％,(8.5±1.8) days vs.(12.1±2.6) days,x2=4.897,5.072,2.351,respectively,all P＜0.05].No significant difference in the rate of return to ICU was found between the two groups(P＞0.05).Conclusions The combination of perioperative Ambroxol and Ipratropium can effectively improve lung function by improving ventilation and gas exchange function,reduce postoperative pulmonary complications and shorten the length of postoperative hospital stay in lung cancer patients aged 70 years and over.

Objective To study the radiofrequency ablation (RA) and vagal denervafion (VD) in surgical treatment of long-standing atrial fibrillation (AF) associated with rheumatic heart disease (RHD).Methods Retrospective analysis the cardiac rhythm by 24-hour Holter monitoring during 5-year follow-up after total Maze procedure accompanied rheumatic mitral valve replacement.Between June 2006 and December 2007,a total of 173 consecutive patients with long-standing AF-associated RHD underwent mitral valve replacement and ablation maze procedure,92 cases had RA alone and 81 had RA + VD.Results Although Kaplan-Meier curve shows that the freedom from AF at 5 years follow-up time were similar(P =0.718),the percentage of antiarrhythmic drug therapy was significant higher in the RA group during early postoperative period(4th month,54.1％ vs.34.7％,P=0.017;5th month,39.2％ vs.21.3％,P=0.018;6th month,23.0％ vs.10.7％,P =0.044),and the percentage of those free by AF was significant lower(6th month,82.2％ vs.93.8％,P =0.023;1st year,76.1％ vs.89.9％,P=0.019).Conclusion Total maze procedure with bipolar radiofrequency ablation is effective to treat longstanding AF associated with rheumatic valve disease.Vagal denervation helped to maintain stable sinus rhythm and lower antiarrhythmic drug therapy at the early stage,but there was no additional benefit after the 1 st year of follow-up,it may be caused from the reactivation of vagal plexus electrical activity.

BACKGROUND:For patients with degenerative mitral annulus dilatation complicated by atrial fibrilation, atrial fibrilation is needed to be treated concomitant with artificial valve ring implantation. Although Cox maze III procedure is the gold standard for treatment of atrial fibrilation, its safety has been questioned. OBJECTIVE:To evaluate the safety of artificial valve rings implantation and Cox Maze III procedure for treatment of degenerative mitral annulus dilatation complicated by atrial fibrilation. METHODS: A total of 43 patients with degenerative mitral annulus dilatation and atrial fibrilation were enroled and were randomly divided into test group (n=21) and control group (n=22). Patients in the test group were treated with mitral valvuloplasty and Cox Maze III procedure. Patients in the control group were only treated with mitral valvuloplasty. RESULTS AND CONCLUSION: (1) Intraoperative safety indexes: the durations of cardiopulmonary bypass and aortic clamping. and the incidence of hemoglobinuria in the test group were higher than those in the control group (P < 0.001). There was no significant difference in the urine output between these two groups. (2) Postoperative safety indexes: the incidence of atrial dysrhythmia (i.e., any atrial dysrhythmia other than atrial fibrilation), proportion of patients requiring a temporary pacemaker and hospital stays in the test group were higher than those in the control group (P < 0.05). There were no significant differences in the secondary-thoracotomy bleeding rate and incidence of stroke between these two groups. (3) Postoperative outcomes: the rates of cardioversion to achieve sinus rhythm at the day of surgery and at discharge in the test group were significantly higher than those in the control group (P < 0.001). At discharge, patients in both groups were al at NYHA functional class Ior II. Echocardiography showed trace to mild mitral regurgitation and complete closure of the tricuspid valve. There were no significant differences in the left atrial diameter, left ventricular diameter, left ventricular volume, and ejection fraction between these two groups. However, the proportions of mitral and tricuspid valve peak A-waves were significantly higher than those in the control group (P < 0.001). These results show that artificial valve ring implantation and Cox Maze III procedure is a safe and effective method for treating degenerative mitral annulus dilatation complicated by atrial fibrilation during operation.

AIM: To explore the mechanism of osthole on elderly spontaneously hypertensive rats. METHODS: 20-month-old spontaneously hypertensive rats (SHRs) and healthy Wistar-Kyoto (WKY) rats were purchased. SHRs were treated with osthole (i.g.) for 8 weeks. The systolic blood pressure and diastolic blood pressure of rats were monitored. Hematoxylin-eosin staining (H&E), periodic acid-schiff staining (PAS) and Masson staining were used to observe the pathological changes of rat kidney tissues. The activity of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) in rat kidney was detected by ELISA kit. PI3K/Akt/mTOR signaling pathway related proteins were detected by western blot. RESULTS: Osthole reduced the systolic and diastolic blood pressure of SHRs, improved the histopathological changes of SHRs kidney, reduced the activity of MDA in SHRs kidney, and increased the activity of SOD and GSH. Osthole reduced the levels of p-PI3K, p-Akt and p-mTOR. CONCLUSION: Osthole reduces the activity of PI3K/Akt/mTOR signaling pathway and exerts a protective effect on renal oxidative stress injury in aged spontaneously hypertensive rats.

Objective To investigate the clinical values of detection of plasma miRNA-544a (miR-544a) expression level in the diagnosis and treatment of lung cancer.Methods A total of 110 patients diagnosed with lung cancer in Ⅰ-Ⅱ stage from June 2017 to March 2018 in Shiyan Taihe Hospital were collected,and 35 patients with benign pulmonary nodules and 64 healthy people were also collected as the controls.Realtime quantitative polymerase chain reaction (RT-qPCR) was used to detect the level of miR-S44a in plasma of 64 healthy people,110 patients with lung cancer (50 newly diagnosed patients without anti-tumor treatment,33 patients one week after radical resection,27 patients after one week chemotherapy with the same dose) and 35 patients with benign pulmonary nodules.Of the 50 newly diagnosed patients,42 cases were non-small cell lung cancer and 8 cases were small cell lung cancer.The plasma expression level of miR-544a in each group was compared by using Mann-Whitney U test,and the efficacy of miR-544a in the diagnosis of lung cancer was evaluated by the receiver operationg characteristic (ROC) curve and the area under the curve (AUC).Results The plasma expression levels of miR-544a in the newly diagnosed untreated lung cancer group,one week after operation group,one week chemotherapy group,healthy control group and benign pulmonary nodule group were 1.40 nmol/L (0.55 nmol/L,8.76 nmol/L),33.52 nmol/L (3.64 nmol/L,250.47 nmo/L),8.87 nmol/L (0.68 nmol/L,125.43 nmol/L),0.31 nmol/L (0.17 nmol/L,1.19 nmol/L),1.04 nmol/L (0.31 nmol/L,4.62 nmol/L),respectively,and the differences between untreated lung cancer group and the other 4 groups were statistically significant (Z =-4.483,P ＜ 0.001;Z =-4.274,P ＜ 0.001;Z =-2.562,P =0.01;Z =-2.152,P =0.031).The expression levels of miR-544a in non-small cell lung cancer group and small cell lung cancer group were 1.40 nmol/L (0.66 nmol/L,8.76 nmol/L) and 1.37 nmol/L (0.26 nmol/L,36.97 nmol/L),respectively.The differences between non-small cell lung cancer group and healthy control group and benign pulmonary nodule group were statistically significant (Z =-4.463,P ＜ 0.001;Z =-2.026,P =0.043).Compared with the healthy people,the AUC of miR-544a for diagnosing the lung cancer was 0.841,the sensitivity was 87.5 ％,and the specificity was 68.0 ％.Compared with the benign pulmonary nodule,the AUC for diagnosing lung cancer was 0.638,the sensitive was 45.7 ％,and the specificity was 80.0 ％.Conclusions The plasma expression level of miR-544a has certain significances in the differential diagnosis of early stage lung cancer and benign pulmonary nodules and healthy people,and it can be used as a potential biomarker for diagnosing early stage lung cancer,especially for the non-small cell lung cancer.The plasma expression of miR-544a is increased after surgery or chemotherapy,suggesting that its expression may be related to the occurrence and development of lung cancer,and miR-544a may become a new target for cancer treatment.