Objective To develop and verify a qPCR method for the qualitative and quantitative analysis of residual host DNA in human rabies vaccine(Vero cells) stock solution.Methods The qPCR standard curve was established by using the Vero cell DNA quantitative national standard,and the residual host DNA was extracted using magnetic beads.The specificity,repeatability,intermediate precision,accuracy and durability of the method were verified,and the linear range and limit of quantification were determined.The residual DNA of three batches of human rabies vaccine(Vero cells) stock solution was quantitatively analyzed and the fragment size was qualitatively analyzed by using this method.Results The correlation coefficients(R~2) of Vero cell DNA quantitative national standard amplification standard curve were all more than 0.99 by qPCR,and the quantitative range was 0.3 pg/mL～30 ng/mL.The method showed good specificity and repeatability.In the verification of intermediate precision,accuracy and durability,the relative standard deviations(RSD)of detection results of the samples were all less than 10%.The residual DNA content of Vero cells in three batches of stock solution was 0.20～0.77 ng/dose,which met the relevant standard of Chinese Pharmacopoeia(Volume Ⅲ,2020edition).The residual DNA fragments greater than 154 bp accounted for 52%～63%.Conclusion The developed qPCR method for the detection of residual DNA in human rabies vaccine(Vero cells) stock solution had good specificity,repeatability,intermediate precision and durability,and qualitatively and quantitatively analyzed the residual DNA rapidly and accurately,which was of great significance for improving the detection and control of residual DNA content in the production process and final product of human rabies vaccine(Vero cells).

Objective To explore the experience of surgical treatment for appendiceal carcinoid.MethodsThe clinical data of 12 appendiceal carcinoid cases received surgery were retrospectively studied.ResultsPreoperatively,all the 12 cases were considered as appendicitis.The definite diagnosis was verified by pathologic study postoperatively.Among the 12 cases, tumors less than 1 cm 9 cases, between 1 cm to 2cm 2cases, but without serosal involvment or lymph node metastasis(2 cases) recieved simple appendectomy.One case with a tumor larger than 2cm and serosal involvment underwent fight hemicolectomy.ConclusionOwing to its lack of specific symptoms,the diagnosis of appendiceal carcinoid was usually reached after operation.The choice of surgical procedures should based on the size and depth of invasion of the tumor and lymph node metastasis status.

Objective The aim of this study was to explore the effects of volume controlled ventilation (VCV),pressure controlled ventilation(PCV)and pressure controlled ventilation-volume guaranteed (PCV-VG)on respiration and circulation in elderly patients undergoing thoracic surgery. Methods Thirty-six elderly patients who underwent thoracic surgery were enrolled in our study. Patients were divided into VCV,PCV and PCV-VG groups according to randomized design. The hemodynamic and respiratory data and the arterial blood gases had been recorded in the pre-operation,20 min,40 min,60 min after OLV and 20 min after the resumption of two lung ventilation. Results Compared with VCV group,Ppeak value was significantly lower in PCV and PCV-VG groups(P < 0.05),and the difference was not found between the PCV and PCV-VG groups. PaO2 levels in PCV and PCV-VG groups were higher than that in VCV group after the point of OLV+40(P < 0.05). Comparison of PCV group,PaO2 in PCV-VG group was higher,but did not show a significantly improved during OLV(P > 0.05). Conclusion Compared with VCV,the use of PCV and PCV-VG have significant advantages in the operative oxygenation and airway pressure for elderly patients undergoing OLV.

Obiective To observe the clinical effect of treating upper respiratory infection induced coughing with atomizated ioint of compound liquorice oral solution and ipratropium bromide.Methods 69 cases of respiratory tract infection induced coughing were randomly recruited into A,B,and C group.Group A was treated with atomizated joint of compound liquorice oral solution and ipratropium bromide;Group C was treated with azithromycin and levofloxaein;Group B was treated with the combined therapy of Group A and Group C.Clinical effects and side effects were observed after the treatment.Results Symptom scores of cough in group A and group B were lower than those in group C.The difference was statistically significant(P＜0.05).Dry mouth and throat complaints disappeared in group A and group A and group B after the treatment.Conclusion Tomizated ioint of compound liquorice oral solution and ipratropium bromide had good therapeutic effects in treating cough induced by upper respiratory infection.

Objective To improve the protective strategy of occupational risks for technicians in blood centers so as to ensure their health and safety.Methods The occupational risk factors were analyzed by the 5W1H method,and the corresponding protective measures were introduced.Then,the protective measures were verified dynamically by the PDCA circulation,and were standardized and further spread.The projects with poor effect were replanned and entered the PDCA recycling.Results The system of the 5W1H method combined with PDCA circulation could identify and control various occupational risks,and effectively prevent the occupational detriments.Condusion Combined application of the 5W1H method with PDCA circulation is an effective way to protect the health and safety of technicians in blood centers and an effective mode to improve the management of occupational risks.

Objective To discuss the perioperative nursing points of vitreoretinal surgery under tempo-rary kerstoprosthesis combined with penetrating keratoplasty for treatment of complex ocular trauma and anteri-or and posterior ocular segment lesion. Methods Clinical data of 21 patients (21 eyes) undergoing vitreo-retinal surgery under artificial cornea combined with penetrating keratoplasty were analyzed, perioperative nursing problems were evaluated and corresponding nursing measures were adopted. Results Visual acuity of 16 eyes were greatly improved,other 5 eyes showed no amelioration.By effective nursing mea-sures, all nursing problems were settled. Conclusions Although the functional outcome of a combined procedure is limited,it contributes to preserve vision of patients. Good periopertive nursing is pivotal for successfal operation.

Objective To construct pcDNA 3.1(+)/HPV 18 E 6 fusion gene and a single-codon mutation pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R fusion gene in eukaryotic expression vector and study the effects on proliferation and apoptosis of cervical carcinoma cell line Hela. Method HPV 18 E6 gene sequence and the single-point mutation HPV 18 E6 F49R or HPV 18 E6 F127R were amplified from total RNA of Hela cell line by reverse transcription- polymerase chain reaction ( RT- PCR), then the gene sequences were respectively inserted into pcDNA 3.1(+) vector to reconstruct recombinant plasmids which were transfected transiently into Hela cells. MTT and RT-PCR were used to test the expression levels of HPV 18 E6 and the growth of HeLa cells after transfected about 48 h. The proliferation and apoptosis of Hela cells were detected respectively by cell counting and AO/EB fluorescent vital staining. Results The pcDNA 3.1(+)/HPV 18 E6, pcDNA 3.1(+)/E6 F49R and pcDNA 3.1(+)/E6 F127R eukaryotic expression vectors were successfully constructed. The gene of HPV 18 E6 was discriminably detected in the HeLa cells which were transfected with the recombinant plasmids. After several days, the proliferation of Hela cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R plasmid were obviously inhibited and the apoptotic rates were significantly increased, then the proliferation of cells transfected with pcDNA(+)/HPV 18 E6 was rather increased slightly, and we could observe the phenomena of early apoptosis and the formation of thekaryopyknosis by fluorescent microscope in the cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R. Conclusion The eukaryotic expressing vectors encoding HPV 18 E6 F49R and HPV 18 E6 F127R provide fundamental basis for the further study on HPV 18 E6 mechanism as well as prevention and treatment of uterine cancer.

Objective: To investigate the effects of rosuvastatin on reactive oxygen species (ROS) production and periostin, cardiotrophin-1 (CT-1) expression, and to explore rosuvastatin on ventricular remodeling in experimental rats after acute myocardial infarction (AMI). Methods: A total of 45 male Wistar rats were randomly divided into 2 group, Sham operation group,n=15 and AMI group,n=30, the AMI model was established by left anterior descending coronary ligation. After 24 hours of AMI, the rats were further divided into 2 groups, AMI + rosuvastatin group, the rats received gastric rosuvastatin 1mg/(kg?d), and AMI group, the rats received gastric normal saline.n=15 in each group and all animals were treated for 6 weeks. The mRNA and protein expressions of CT-1 and periostin were examined by real time RT-PCR and immunohistochemistry, the contents of superoxide anion (O2-·) and hydroxy radical (OH·) were detected by colorimetric method among different groups. Results: Compared with Sham operation group and AMI + suvastatin group, the mRNA and protein expressions of CT-1, periostin, the contents of (O2-·), (OH·) and left heart weight index were increased in AMI group at non-infraction zone,P<005. Compared with Sham operation group, the mRNA and protein expressions of CT-1, periostin, the contents of (O2-·), (OH·) and left heart weight index were increased in AMI + suvastatin group at non-infraction zone,P<005. Compared with AMI group, the mRNA and protein expressions of CT-1 and periostin were decreased in AMI + rosuvastatin group,P<005. Conclusion: Rosuvastatin may improve ventricular remodeling via inhibiting ROS production and CT-1, periostin expression in experimental rats after AMI.

Objective To study the clinicopathologic characteristics and the prognostic factors of gastric stump cancer (GSC). Methods A total of forty-seven patients with GSC from Jan 2000 to Dec 2006 were enrolled in this study for retrospective analysis. Initial surgery was performed for gastric benign disease in 39 patients and for malignant disease in 8 patients, which were divided into 2 groups for analysis. The prognosis of all 47 patients were analyzed. Results The mean interval between previous gastrectomy and diagnosis of GSC was 24.4 years. Tumor developed mostly in the patients with Billroth- Ⅱ reconstruction, and male more than female. Tumor located at anastomotic site mostly, at stump stomach and cardia secondly. The mean interval for patients who had undergone their first gastrectomy for malignant disease was shorter than that with benign disease(P＜0.05). Histology, therapy and prognosis showed no significant differences between two groups (P＞0.05). Disease TNM stage and total radical gastrectomy were shown to be significant predictor for the outcome of patients with GSC (P ＜0.01). Conclusion Now the GSC patients with initial surgery performed for malignant disease are increased, which are no siginificant different to patients with benign disease. Early diagnosis and an aggressive surgical approach are crucial to achieve better outcomes for patients with GSC.

Objective:To investigate the door-to-balloon (D2B) time and its influencing factors for Percutaneous Coronary Intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI).Methods:180 cases of patients with STEMI in our hospital from January 2014 to April 2016 were selected.PCI therapy were operated on all patients after their consent.The pre-hospital delay time and D2B time of the patients were recorded.The related information of the patients,including demographic data,clinical factors,background of the disease and psychological factors,were investigated by the questionnaire survey.The patients were divided into short D2B group (D2B time≤ 126 min,n=96) and long D2B group (D2B time＞126 min,n=84).Univariate and multivariate logistic regression methods were used to analyze the influencing factors of D2B time.Results:The median D2B time of all the patients was 126 min,and only 26.7％ of patients' D2B time controlled within 90 min.Univariate analysis showed that differences of sudden attack,pay attention to symptoms,someone was present when attack,symptoms progress was fast,in hospital during holiday,no symptom in CCU,outpatient treatment,transfered by emergency medical service system (EMSS),time in CCU (6 am-10 pm),angina before infarction and pre-hospital delay time between the two groups were statistically significant (P＜0.05).Multivariate logistic regression analysis showed that in hospital during holiday,outpatient service,no symptom in CCU,pay attention to symptoms,use of transfered by EMSS,time in CCU (6am-10pm) are the factors affecting the time of D2B (OR=2.62,2.04,1.59,0.52,0.28,0.61 P＜0.05).Conclusion:The D2B time of most patients with STEMI can not reach the guidelines.The factors of patients,doctors,accepting mechanism of hospital are all related with D2B time.