Objective To develop and verify a detection method for recombinant E.coli plasmid DNA(pDNA) copy number in the host based on qPCR technology platform, so as to provide a reliable detection method for the development of mRNA vaccine. Methods According to the Top10 genomic DNA sequences of E.coli, the primers and probes were designed for qPCR amplification, and the primer concentration was optimized. The whole DNA(host DNA + pDNA) of recombinant E.coli/pUC57-HA was extracted and used as the template to develop a qPCR method for the detection of pDNA copy number. The linear range, specificity and reproducibility of the method were verified. The recombinant E.coli/pUC57-HA was cultured at37 ℃ for 24 h, and the samples were taken every 2 h and detected for the pDNA copy number by the developed method.Results The optimum primer concentration(F/R) was 0. 5 μmo/L. The whole DNA of recombinant E.coli/pUC57-HA showed a good linear relationship with Ct values in the dilution range of 10~1-10~5times, each R~2= 1. 00. The amplification results of E.coli/pUC57-HA and E.coli Top10 were positive, and the negative control(ddH_2O) showed no amplification curve. The CV of three repeated detections for pDNA copy number of recombinant E.coli/pUC57-HA was less than 5%. The pDNA copy number of E.coli/pUC57-HA was stable during the incubation period of 0-6 h, decreased during 6-12 h, increased during 12-16 h,and stabilized again during 16-24 h. Conclusion The developed qPCR method has good specificity and reproducibility, which can be used to detect the copy number of pDNA in recombinant E.coli host, and effectively monitor the change of pDNA content during culture.

Atrial fibrillation (AF) is the most common clinical arrhythmia.Atrial electrical remodeling and structural remodeling are the important mechanisms for AF.The intracellular Ca2+ handling abnormalities play an important role in the induction of triggered ectopic activity and in the activation of Ca2+-related cell signaling which mediates fibrillatory remodeling.In addition,the importance of microRNAs,which are a new class of small noncoding sequences that regulate gene expression,has been found in both electrical and structural remodeling.The new discovery of AF mechanisms are helpful to exploring the effective treatment for AF,showing good prospects for the use of translational perspectives.

Objective To discuss the efficacy of sufentanil combined with propofol in iv anesthesia during laparoscopic cholecystectomy.Methods Totally 182 patients received laparoscopic cholecystectomy.They were divided into two groups according to odd or even number of hospitalization.The control group (81 cases) were given remifentani combined with propofol for iv anesthesia.The observation group (101 cases) were given sufentanil combined with propofol for iv anesthesia.The anesthetic effect of sufentanil combined with propofol for iv anesthesia during laparoscopic cholecystectomy was evaluated by breathing recovery time,opening time,extubation time,VAS scores after 1,6,12 h,and complications during anesthesia.Results Breathing recovery time,opening time,and extubation time of observation group were longer than those of control group (P ＜ 0.05).The VAS scores after 1,6,and 12 h of observation group were lower than those of the control group (P ＜ 0.05).During anesthesia,the proportion of restlessness in observation group was lower than that in control group (P ＜ 0.05).The complication rate of observation group was lower than that of control group (P ＜ 0.05).Conclusion Sufentani combined with propofol has a good anesthetic effect on patients receiving laparoscopic cholecystectomy with mild postoperative pain and less restlessness.It is worthy of clinical use.

The rehabilitation therapeutic method of dysphagia after stroke included movement therapy and compensatory technology (including neurodevelopmental therapy), neuromuscular electrical stimulation, surface electromygraphy biofeedback training, non-invasive brain stimulation, balloon ductal dilatation technique, acupuncture and moxibustion, drug, and surgical treatment. The movement therapy and compensatory technology were the basis of therapy and the upward and forward movements of tongue-throat complex were the core of therapy. The combined treatments of early, initiative, individual, and various methods were emphasized, while the nutrition and the preven-tion and cure of aspiration pneumonia of patients should be paid attention to.

Objectives: To observe the radiological and pathological changes of flow diverting sent implanting into abdominal aortae of rabbits at 3 months and to evaluate the safety of stents for normal arteries of rabbits. Methods: The abdominal aorta beyond the left renal artery opening was showed via preoperative angiography. The flow diverting stents with metal coverage of 35% to 45% were implanted into the abdominal aortae of 20 New Zealand white rabbits. Intravenous digital subtraction angiography was performed at 4 weeks after the procedure. Arterial digital subtraction angiography was performed for all the animals at 3 months and they were sacrificed for histopathological examination. The angiography showed that the diameter changes of abdominal aortae on the stenting sites at different time points were measured and compared. The relationship between the neointima and the metal coverage of local stent were analyzed. Results: The mean diameter of the abdominal aortae before stenting, immediately after stenting, at 4 weeks, and at 3 months were 3.47 ± 0.44 mm, 3.65 ± 0.40 mm, 3.57 ± 0.36 mm, and 3.51 ± 0.30 mm, respectively. There were significant differences among the different time points (P=0.009). The neointima fully covered the stents with a mean thickness of 95 ± 42 μm. They were predominantly composed of smooth muscle cells, collagen fibers, and a small amount of macrophages. The neointimal thickness was positively correlated with the local coverage of the metal stents (r = 0.523; P = 0.001). Conclusion Three months after the implantation of flow diverting stents, the abdominal aorta intima thickened slightly, and the stents were safety for normal arteries of rabbits.

Objective To investigate the influence of ziprasidone combined with amend electric shock on the efficacy and metabolism of patients with first-episode schizophrenia.Methods According to random number table,86 cases with first-episode schizophrenia from July 2013 to July 2016 in the Second Affiliated Hospital of Xinxiang Medical University were chosen and divided into observation group and control group.The patients in observation group were given ziprasidone combined with first-episode schizophrenia,the patients in control group were taken with olanzapine combined with first-episode schizophrenia.The PANSS total score,CGI score,total efficacy,adverse reaction,body mass index,glycometabolism,and lipid metabolism of two groups were observed and compared.Results The PANSS score,CGI score of two groups were all obvious decreased,which had no significant difference,the total efficacy of two groups had no significant difference;The TESS scores were all decreased,and the TESS score of observation group was obviously lower than that of control group (P ＜ 0.05).Compared with before treatment,the BMI,fasting blood sugar,triglyceride,total cholesterol,high/low density lipoprotein and fasting insulin level of observation group after treatment had no obvious significant difference,while the BMI,fasting blood sugar,triglyceride,total cholesterol,low density lipoprotein,and fasting insulin levels of control group had obvious significant difference,and the BMI,fasting blood sugar,triglyceride,total cholesterol,and fasting insulin of observation group were obviously lower than control group (P ＜ 0.05).Conclusion Olanzapine and ziprasidone combined with amend electric shock have obvious efficacy on patients with first-episode schizophrenia,while the ziprasidone had lower adverse reaction and metabolic side effects,which is worth of clinical promotion and application.

Objective To determine the content of cholesterol in Gekko Gecko.Methods High performance liquid chromatography was performed on Hypersil ODS2 (4.6 mm?150 mm,5 ?m).The chromatorgraphic conditions were as follows:methanol as mobile phase,flow rate being 1.0 mL/min and detecting wavelength at 208 nm.The column temperature was 40 ℃.Results A good linearity of cholesterol was shown in range of 0.885～14.000 ?g (r=0.999 5),the average recovery was 99.65%,RSD=3.45% (n=6).Conclusions This method is simple,rapid and accurate,and may provide a reliable way for the quantification of cholesterol in Gekko Gecko.

Objective To study the expressions and significances of matrix metalloproteinases-3 (MMP-3) and tissue-inhibitor of matrix metalloproteinase-3 (TIMP-3) in transitional cell carcinoma of bladder.Methods Immunohistochemistry was used to detect the expressions of MMP-3 and TIMP-3 in 48 cases of transitional cell carcinoma of bladder and 20 normal bladder tissues.Results MMP-3 and TIMP-3 in normal bladder tissues did not express.The more grading and staging,the more MMP-3 expression,but no statistical significance(P ＞0.05),by contrast,TIMP-3 expression decreased and has statistically significant in the tumor grading (P ＜ 0.05).Conclusions High expression of MMP-3 and low expression of TIMP-3 is related to the malignancy of transitional cell carcinoma of bladder and may play an important role in the development of transitional cell carcinoma of bladder,as an important reference to judge the degree of malignancy of transitional cell carcinoma of bladder and prognostic indicator.

Objective To explore the microsurgical treatment of the fourth ventricular tumors. Methods A total of 29 cases of the fourth ventricular tumors were given microsurgical treatment from January 2000 to June 2003 in this hospital. In 5 cases the tumor was located close to the cortex with the superior vermis involvement and the microsurgical operation was performed through the cerebellar vermis. The remaining 24 cases underwent microsurgical resection of tumor through the cerebellomedullary fissure approach. Results A total resection was achieved in 23 cases, and a subtotal resection in 6 cases. Postoperative complications included 2 cases of hemorrhage of the upper digestive tract, 1 case of mutism, 1 case of hematoma of the fourth ventricular, 1 case of epidural hematoma remote from surgical site, and 1 case of irregular breath. Follow-up surveys in 19 cases for 3~12 months (mean, 5.5 months) revealed no tumor recurrence. Conclusions Preoperative proper differentiating characters of tumors, clarifying the position of tumors' basal part, selecting appropriate surgical approach, and understanding microanatomy of the fourth ventricular are the key to successfully complete the procedure.

Objective To systemically evaluate the quality of randomized controlled trials(RCTs) of recurrent oral ulcer(ROU) treated by traditional Chinese medicine.Methods The randomized controlled trials of TCM treatment on ROU were retrieved,and the related information from database was retrieved and assessed with Cochrane Handbook.Results Fifty-three articles in 41 journals were found.There were 12 diagnostic criteria and 8 effect criteria.Base-line comparision in 39 articles,blind methods in 3 articles,follow-up survey in 2 articles,adverse reaction in 10 articles,no case loss in all articles were reported.Conclusions The quality of RCTs of TCM for ROU should be improved.