Objective To investigate the inhibitory effect of siRNA human telomerase transcriptase (hTERT) on activity of telomerase and proliferation of lung carcinoma cell A549. Methods A plasmid including U6 promoter and siRNA of hTERT was designed and constructed. The plasmid was transfected into A549 cell line. The telomerase activity was tested by telomerase repeat amplification protocol ELISA (TRAP-ELISA). MTT assay was used to assay the cell proliferation activity,and hTERT expression was assessed by Western blot. Result The U6 expression plasmid that was constructed for hTERT gene 745 showed obvious interfering effect. hTERT-siRNA could down-regulate the expression of hTERT protein,inhibit telomerase activity and proliferation of A549 cells. Conclusion siRNA of hTERT can inhibit the expression of human telomerase and proliferation of A549 cells. It may open a new approach to the use of siRNA as a new tool to study gene function in cancer cell lines,and may be developed to be a new gene therapeutic agent for cancer.

Objective To investigate the predictive value of 4 183 Da peptide of dermcidin protein in the early diagnosis and differential diagnosis of ischemic heart disease.Methods A prospective controlled study was conducted.Serum samples were drawn from 161 patients with acute coronary syndrome [ACS,including 46 patients with unstable angina (UA),23 with acute non-ST elevation myocardial infarction,and 92 with acute ST segment elevation myocardial infarction],111 subjects for routine physical examination,including 45 patients with hypertension history,42 with coronary heart disease,22 with diabetes,and 54 patients with non-ACS (including pulmonary embolism,aortic dissection aneurysm,arrhythmia,myocarditis,coronary myocardial bridge,pleurisy,pneumothorax,pneumomediastinum,rib fracture,reflux esophagitis,peptic ulcer,and pancreatitis) to serve as controls.4 183 Da peptide of dermcidin protein was assessed with matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technology,and myeloperoxidase [MPO,determined by point-of-care testing (POCT) and enzyme linked i mmunosorbent assay (ELISA),respectively],high sensitive C-reactive protein (hs-CRP),heart type fatty acid binding protein (H-FABP),myoglobin (MYO),cardiac troponin Ⅰ (cTnⅠ),and MB isoenzyme of creatine kinase (CK-MB) were quantitated with biochemical analysis.The power of the biomarkers above for early diagnosis and differential diagnosis for ischemic heart disease were judged by comparison of their sensitivity and specificity.Results ① It was showed by one-way ANOVA that 4 183 Da peptide was higher in ACS group than that in control group (relative abundance:22.05 ± 16.97 vs.15.52 ± 14.09,P =0.001),but no difference was found between ACS group and non-ACS group (relative abundance:22.05 ± 16.97 vs.19.99 ± 17.63,P =0.416).② The specificity and sensitivity of the 4 183 Da polypeptide and MPO for predicting ACS and UA were compared with the receiver operating characteristic curve (ROC).It was showed that the 4 183 Da polypeptide had predictive values for ACS and UA,and the areas under the ROC curve (AUC) was 0.625 and 0.651 (both P ＜ 0.01),but MPO was not found to have predictive value (AUC was 0.440 and 0.336,respectively,both P ＞ 0.05).③ It was showed by the values of multi-markers in differential diagnosis of ACS and non-ACS disease that the specificity and sensitivity of 4 183 Da peptide in the differential diagnosis of acute myocardial infarction (AMI) and non-ACS disease were less than those of MYO,cTnⅠ,H-FABP,markers of myocardial damage,which AUCs were 0.569 vs.0.796,0.833,0.838,and equal to MPO (POCT/ELISA) and hs-CRP,AUC of which was 0.569 vs.0.505 (POCT)/0.477 (ELISA) and 0.545.But both the value of 4 183 Da peptide and MYO,cTnⅠ,H-FABP in the differential diagnosis of UA and non-ACS disease was not found,where AUC was 0.456,0.525,0.658,0.568.Conclusion 4 183 Da polypeptide,a fragment of dermcidin protein,may have association with the onset of ischemic heart disease,and may be helpful in the early diagnosis of ACS.

Objective To investigate the epidemic patterns and the characteristics of influenza in Chi-na through a Meta-analysis based on the studies published in domestic literatures.Methods Related articles published during 2005 to 2012 were screened out from domestic databases and analyzed through a Meta-analysis with Review Manager 5.0 software.Results Twenty-two articles covering 957 901 patients with influenza-like-illness (ILI) and 148 233 pathogen samples were screened out according to the inclusion criteria.No significant difference with the ILI diagnosis rate was found between subjects at age 0-4 years and those at age 15-59 years. Higher ILI diagnosis rates were observed in those two groups as compared with subjects elder than 60 years old. Most of the pathogen samples were carried by subjects aged 25-59 years.More influenza virus strains were isola-ted in 2009 as compared with those of the seven other years (OR=2.25, 95%CI=1.27-3.70).There was sta-tistical difference between the numbers of influenza A H1N1 and seasonal influenza A strains (OR=2.25, 95%CI=1.30-3.91) .Significant difference was also observed between the numbers of influenza A and influenza B strains (OR=4.05, 95%CI=2.53-6.47).Conclusion There was significant difference with the diagnosis rate between subjects aged 0-4 years and those aged≥60 years.More attention should be paid to people at high risk of infection (0-4 years old and≥60 years old) and those at 25-29 years with high mobility and social inter-course for the timely prevention and control of pandemic influenza.The detection rate of influenza virus strains was increased during the outbreak of novel influenza A H1N1 infection in 2009.After that outbreak, the detec-tion rate of novel influenza A H1N1 strains was 2.25 times the rate of seasonal influenza strains.The detection rate of influenza A was 4.05 times the rate of influenza B virus strains.Therefore, it is necessary to strengthen the surveillance for influenza A virus and other epidemic influenza virus strains.

Objective To analyze the profile of dermcidin (DCD) changes in different stages of acute coronary syndrome (ACS) by quantifying the serum 4 183Da DCD peptide fragment deriving from different ACS patients treated with early antithrombotic therapy.Methods A total of 118 patients with confirmed diagnosis of ACS were enrolled. Immediately after visiting a doctor, the venous blood was collected and afterwards instantly the patient was given orally 300 mg of aspirin and 300 mg clopidogrel, and according to the patient's condition and the consent of his/her or acknowledgement of family members achieved, emergency percutaneous coronary interference (PCI) or thrombolysis or conservative treatment was adopted separately. After anti-thrombotic treatment, at 2, 4, 6, 8, 10, 12, 16, 20, 24, 32, 40, 48, 60 and 72 hours, venous blood was collected and serum isolated respectively. The concentration of 4 183Da DCD fragment in serum was determined by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Simultaneously, the myoglobin (Myo), cardiac troponin I (cTnI) and MB isoenzyme of creatine kinase (CK-MB) were also detected.Results The mean relative strength of nature logarithmic transformations of 4 183Da DCD fragment of 118 patients with ACS was 2.75±1.02 before treatment on admission, and after intervention therapy (mainly antithrombotic therapy) it was decreased to 1.84±1.19 (P = 0.005) and 1.74±1.12 (P = 0.000) at 2 hours and 4 hours, respectively, and then after 4 hours it was slightly elevated. 4 183Da polypeptide increased earlier than myocardial injury markers.Conclusion Aspirin and clopidogrel can significantly decrease the concentration of 4 183Da DCD peptide fragment in serum in patients with ACS, which indicates that the DCD fragment could be used as one of the indexes for observation on early efficacy of antithrombotic therapy.

Objective To observe the clinical efficacy of Yuganlong capsule combined with entecavir in treatment of liver fibrosis in patients with chronic hepatitis B cirrhosis. Methods A total of 57 cases of patients with chronic hepatitis B cirrhosis were randomly divided into treatment group(30 cases given entecavir) and control group(27 cases treated with Yuganlong capsule combined with entecavir).Fibroscan values,hepatic fibrosis indexes(HA,LN,C-Ⅳ,PC-Ⅲ),liver function index(ALT,AST,GGT,T-BiL),and ultrasonic imaging index(inner diameters of portal vein,thickness of spleen) were detected. Results Before treatment, all indicatorshad no significant differences between the two groups(P>0.05).After 96 weeks of treatment,the Fibroscan value,HA, LN,C-Ⅳ,ALT,AST and portal vein diameter were significantly reduced in the treatment group as compared with those before treatment(all P<0.05),and there were significant differences between the treatment group and the control group after the treatment (all P<0.05). Conclusion The effect of Yuganlong capsule combined with entecavir in the treatment of chronic hepatitis B liver cirrhosis is better than that of using entecavir alone to improve liver function and liver fibrosis.

Objective To determine the occurrence and clinical characteristics of ectopic pituitary adenoma (EPA) in China.Methods The study are done by searching systematically and comprehensively on major Chinese and English medical literature databases and conference papers before 2015,which are only limited to collected,summarized,sorted and analyzed EPA cases that reported by Chinese authors in English or Chinese occurred in China.Results ① Among the 86 Chinese articles and 27 English articles related to ectopic pituitary adenoma (EPA) gathered by the author,except for cases that have been confirmed as repeated reports,a total of 73 EPA cases were found.Of 70 cases with complete data,31 were male cases,accounting for 44.29％;39 were female cases,accounting for 55.71％;the ratio is 1:1.3.The frequency of EPA occurrence according to the location of the lesion,from high to low in turn,was sphenoid sinus (31 cases,42.47％),nasopharynx (7 cases,9.59％),suprasellar region (7 cases,9.59％),clivus (5 cases,6.85％),followed by the third ventricle,sphenoid sinus/clivus,nasal cavity,and the temporal lobe of the brain,with the same incidence of 4.11％.According to the functional properties of EPA,the frequency of different secreting hormones adenoma was PRL-ma(21 cases,28.77％),NF-ma (21 cases,28.77％),ACTH-ma (15 cases,20.55％),GH-ma (10 cases,13.70％),TSH-ma (2 cases,2.74％) and FSH-ma (1 cases.1.37％).Three cases of EPA were uncertain in their property due to lack of information.The incidence of PRL and nonfunctional tumors was the highest,which was different from what was reported in other countries.Among them,one case of EPA was in pineal region and one in parapharyngeal space,which was even more rare and were never reported abroad.(② Except for 3 cases with incomplete medical records,15 out of 70 cases of EPA were accompanied by empty sella,accounting for 21.43％,among which 11 (73.33％) cases involved the sphenoid sinus,and 3 (20％) cases involved clivus.The sphenoid sinus and clivus cases together accounted for 93.33％.(③ 29 out of the 69 cases of EPA with complete record were invasive pituitary adenomas,accounting for 42.03％ and including 1 case of pituitary adenocarcinoma,which accounted for 1.45％.(④ All cases were treated with surgery as the first choice,and some of them were treated with radiotherapy or drug therapy.Conc lusion Ectopic pituitary adenoma is extremely rare.By the end of 2015,the total number of cases reported in China is only 73,which are mostly located in the sphenoid sinus,suprasellar region and nasopharynx.In the functional classification,the most common types are still PRL adenoma and nonfunctional adenoma as in situ pituitary adenoma.42.03％ of EPA adenomas are invasive.Surgical resection of EPA is the first choice and some cases can be treated with radiotherapy and drug therapy.

Objective To analyze how enterovirus D68 (EV-D68) protease 2A affects the anti-vi-ral interferon typeⅠ(IFN-Ⅰ) pathway in 293T cells following infection. Methods Western blot was used to detect the expression of recombinant protease 2A, IFN-α and signal transducers and activators of tran-scription 1 (STAT1) at protein level. Expression of EV-D68 viral protein (VP1) and protease 2A was ana-lyzed by immunofluorescence at different time points. Cytopathic effects were recorded to calculate 50％ cell culture infective dose ( CCID50 ) . Expression of the genes involved in the anti-viral IFN-Ⅰ pathway was measured by real-time PCR (RT-PCR). Results The recombinant plasmid pCLIPf-2A was successfully constructed and the expression of recombinant protease 2A could be detected by Western blot 24 h after transfection. The recombinant protease 2A promoted the proliferation of EV-D68 at the late stage of infection and induced the production of IFN-α. Expression of the genes involved in the anti-viral IFN-Ⅰ pathway at mRNA level was up- or down-regulated to different degrees with various trends in different groups following infection. Expression of STAT1 was enhanced in all groups. Conclusions EV-D68 protease 2A promoted the activation of anti-viral IFN-Ⅰpathway in response to viral infection and enhanced the proliferation of virus at the late stage of infection.