OBJECTIVE:To validate the determination method of the content of Morphine in Compound Glycyrrhiza oral solution. METHODS: Samples were detected by solid phase extraction-HPLC on Kromasil C8 column (250 mm?4.6 mm,5 ?m) with mobile phase consisted of 0.05 mol?L-1 KH2PO4-0.0 025 mol?L-1 C14H15NaO3S?2H2O-acetonitrile using gradient elution.The detection wavelength was 220 nm and the flow rate was 1.0ml.min-1.RESULTS:The linear range of morphine was 2.02～20.2 ?g?mL-1(r=0.999 7),and the average recovery was 99.0%(RSD=0.40%,n=9).CONCLUSION: The method is accurate,sensitive and stable,and accurate and reliable in determination results.

Objective To investigate the risk factors of cervical esophagogastric anastomotic fistula after esophagectomy of esophageal cancer.Methods The retrospective case-control study was conducted.The clinicopathological data of 956 patients who underwent esophagectomy and cervical esophagogastrostomy from January 2012 to December 2016 in the First Affiliated Hospital of Zhengzhou University were collected.Patients underwent Sweet or Mckeown surgery.Observation indicators:(1) intra-and post-operative situations;(2) the risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the esophagogastric anastomotic stenosis of patients up to February 2017.Measurement data with normal distribution were represented as the (x)±-s.Univariate analysis and comparison of count data were done using the chi-square test or Fisher exact probability method.Multivariate analysis was done using the Logistic regression model.Results (1) Intra-and post-operative situations:all the 956 patients underwent successful operations,including 107 with Sweet operation and 849 with Mckeown operation.Of 956 patients,336 received thoracotomy and 620 received thoracoscopic surgery.Tumors located in upper,middle and lower esophagus were respectively detected in 143,627 and 186 patients.Operation time,volume of intraoperative blood loss and number of lymph node dissected in 956 patients were (274 ± 67) minutes,(210 ± 167) mL and 18 ± 11,respectively.Of 956 patients,117 had cervical esophagogastric anastomotic fistula,with an incidence of anastomotic fistula of 12.24％ (117/956).Of 117 patients with cervical esophagogastric anastomotic fistula,2 had early stage fistula,110 had middle stage fistula and 5 had later stage fistula;12 were cured by two-tube method (stomach tube and nutrition tube),24 were cured by three-tube method (stomach tube,nutrition tube and chest tube or mediastinal tube),43 were cured by open neck incision dressing,15 were cured by fistula cavity drainage and 17 were cured by esophageal stent implantation.Sixteen patients died in hospital postoperatively,including 6 with cervical esophagogastric anastomotic fistula and 10 without cervical esophagogastric anastomotic fistula.Duration of hospital stay of 956 patients was (16± 11)days,and durations of hospital stay of patients with and without cervical esophagogastric anastomotic fistula were (39± 19) days and (13±6) days.Postoperative pathological examinations:873,9 and 74 patients were respectively diagnosed with squamous cell carcinoma,adenocarcinoma and other types of cancer.TNM staging:stage 0,Ⅰ,Ⅱ,Ⅲ,Ⅳ and unidentified stage were respectively detected in 135,110,325,376,1 and 10 patients.(2) The risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy:univariate analysis showed that gender,age,history of diabetes,surgical method,tubular stomach production,operation time,postoperative pulmonary infection and postoperative aspirating sputum through fiberbronchoscope were risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (x2 =4.179,6.174,4.427,4.377,6.266,7.057,55.036,51.806,P＜ 0.05).Multivariate analysis showed that tubular stomach production,postoperative pulmonary infection and aspirating sputum through fiberbronchoscope were independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (OR =1.922,2.907,2.323,95％ confidence interval:l.203-3.070,1.682-5.023,1.235-4.370,P＜0.05).(3) Follow-up situations:908 of 956 patients were followed up for 2-62 months,with a median follow-up time of 28 months.During the follow up,21 of 111 patients with cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,59 of 797 patients without cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,showing a statistically significant difference in cervical esophagogastric anastomotic stenosis (x2-16.803,P＜0.05).Conclusion Tubular stomach production,postoperative pulmonary infection,postoperative aspirating sputum through fiberbronchoscope are independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy.

Objective To investigate the utilization of ultrasound monitoring the renal blood flow during extracorporeal membrane oxygenation (ECMO).Methods Twentry one cases,who received veinartery ECMO for heart failure,were examinated by bed-side ultrasound before the ECMO initiated,right after the ECMO initiated,each day after the ECMO initiated,and right after the ECMO weaned.The renal interlobar artery peak velocity (Vmax) was measured,and the renal interlobar artery resistant index (RI) was calculated,as well as the values of the serum creatinine (SCr) and blood urea nitrogen (BUN) were recorded.All the data were compared.Results Compared to the variables right after the ECMO initiated,thc Vmax incrcascd (P ＜ 0.05) two days after ECMO initiated and right after the ECMO weaned,while RI (P ＜ 0.05),SCr (P ＜ 0.05) and BUN (P ＜ 0.05) decreased,there being significant differences (P ＜ 0.05).Conclusions While treating patients with extracorporeal membrane oxygenation,the ultrasound can monitoring the renal blood flow effectively,and provide important parameters for the clinical doctors as the basis of the diagnosis and treatment.

Objective To explore the application value of the fusiform tube stomach in the digestive tract reconstruction after thoracoscopic and laparoscopic radical resection of esophageal carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 96 patients with thoracic esophageal cancer who were admitted to the First Affiliated Hospital of Zhengzhou University between November 2016 and May 2017 were collected.All the patients underwent thoracoscopic and laparoscopic radical resection of esophageal carcinoma,45 using thin tubular stomach and 51 using fusiform tube stomach for digestive tract reconstruction were respectively allocated into the tubular stomach group and fusiform stomach group.Observation indicators:(1) intra-and post-operative situations;(2) postoperative complications;(3) detection of gastric hemodynamics;(4) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to November 2017.Measurement data with normal distribution were represented as-x±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the rank sum test.Ordinal data and categorical variables were respectively done using the independent-sample Wilcoxon rank-sum test and chi-square test.Paired experimental data were analyzed by the Friedman test and Wilcoxon test.Results (1) Intra-and post-operative situations:96 patients underwent successful thoracoscopic and laparoscopic radical resection of esophageal carcinoma.The number of closers,time of postoperative mediastinal tube removal,thoracic stomach anteroposterior diameter by postoperative CT examination,number of slices at maximal width of thoracic stomach by postoperative CT examination and duration of hospital stay were respectively 4 (range,3-5),14 days (range,11-45 days),28.35 mm (range,9.96-75.70 mm),0.56 (range,0.33-2.13),16 days (range,12-62 days) in the tubular stomach group and 4 (range,2-4),12 days (range,10-16 days),45.80 mm (range,17.36-89.77 mm),1.10 (range,0.47-2.15),14 days (range,12-61 days) in the fusiform stomach group,with statistically significant differences between groups (Z=4.525,4.240,-3.796,-4.604,2.154,P＜0.05).(2) Postoperative complications:cases with postoperative cervical anastomotic fistula,thoracic gastric fistula,grading Ⅰ-Ⅱ and Ⅲ-Ⅳ of Clavien-Dindo classification were respectively 4,5,32,13 in the tubular stomach group (1 with a combination cervical anastomotic fistula and thoracic gastric fistula) and 0,0,47,4 in the fusiform stomach group,with statistically significant differences between groups (x2 =9.937,7.266,P＜0.05).Patients with complications were improved by symptomatic treatment.(3) Detection of gastric hemodynamics:hemodynamic values of gastric antrum,gastric body and gastric fundus that was detected by non-contact laser Doppler line imaging were respectively 314 PU (range,294-320 PU),252 PU (range,242-259 PU),206 PU (range,194-223 PU) in self-control status of 7 patients and 295 PU (range,277-314 PU),255 PU (range,244-267 PU),219 PU (range,199-233 PU) in tubular stomach model and 277 PU (range,263-300 PU),216 PU (range,201-235 PU),199 PU (range,176-207 PU) in fusiform stomach model,with statistically significant differences among groups (x2 =10.286,14.000,10.286,P＜0.05).There were statistically significant differences in the hemodynamic values of gastric antrum,gastric body and gastric fundus between self-control status and fusiform stomach model (Z=-2.028,-2.384,-2.197,P＜0.05),between self-control status and tubular stomach model (Z =-2.371,-2.371,-2.201,P＜0.05) and between fusiform stomach model and tubular stomach model (Z =-2.201,-2.366,-2.366,P＜0.05).(4) Follow-up situations:among 96 patients,90 were followed up for 6-12 months,with a median time of 8 months.During the follow-up,1 patient in the tubular stomach group died of tumor recurrrence,and no patient died in the fusiform stomach group,with no statistically significant difference between groups (x2 =1.264,P ＞ 0.05).Conclusion Compared with the thin tubular stomach,the fusiform tube stomach can reduce the incidences of postoperative fistula and pulmonary complications and shorten duration of hospital stay after the thoracoscopic and laparoscopic radical resection of esophageal carcinoma,and hemodynamics of the fusiform tube stomach is superior to that of thin tubular stomach.

Objective To establish the assay method for the total polysaccharide in Pudi Enema.Methods Phenol-sulfuric acid method was used for chromogenic reaction.The content of total polysaccharide was measured by UV spectrophotometry at 488.8 nm.Results The total polysaccharides calibration curve was at the range of 0~22.635 mg/L, with regression function being Y=0.062 06 X-0.003 34(r=0.999 8).The recovery of calycosin was 98.36%(RSD=2.34%).Conclusion This method is sensitive,rapid,accurate and reliable.It can be used to assay the content of total polysaccharide in Pudi Enema.

Objective:To analyze differentially expressed microRNAs (miRNAs) and target genes in human respiratory epithelial cells (16HBE) after enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) infection using high-throughput sequencing.Methods:TargetScan and miRDB databases were used to predict the target genes of miRNAs that were both up-regulated or down-regulated after EV71 and CVA16 infection. The genes that were both up-regulated and down-regulated were screened out. GO and pathway analysis of the target genes were conducted to screen immune-related target genes and their corresponding miRNAs. The target genes and their corresponding miRNAs that were up-regulated or down-regulated in both immune-related GO and pathway were further screened. Some miRNAs and their target genes were selected for qRT-PCR verification.Results:There were 598 target genes of up-regulated miRNAs and 1 311 target genes of down-regulated miRNAs and 62 target genes that might be up-regulated or down-regulated simultaneously were screened out. The number of up-regulated target genes involved in immune-related GO and pathway were 17 and 13, respectively, and the number of corresponding miRNAs were 15 and 17, respectively. There were 58 and 47 down-regulated target genes involved in immune-related GO and pathway, respectively, and the number of corresponding miRNAs were 30 and 42, respectively. Three up-regulated target genes were involved in both immune-related GO and pathway and regulated by four miRNAs. Nine down-regulated target genes were involved in both immune-related GO and pathway and regulated by 13 miRNAs.Conclusions:This study was conducive to elucidate the host-pathogen interaction after EV71 and CVA16 infection, and provided reference for studying the pathogenesis of hand, foot and mouth disease.

Objective:To investigate whether coxsackievirus A 16 (CVA16) infection would affect the expression of N6-methyladenosine (m 6A) methylation-related proteins in human bronchial epithelial cells (16HBE), ICR suckling mice and SCRBA2 humanized mice and influence their subcellular localization. Methods:CVA16 was used to infect 16HBE cells at a multiplicity of infection (MOI) of 0.1 and mice at 10 7 CCID 50/ml. Changes in the expression of methyltransferases, demethylases and methylated reading proteins were analyzed by Western blot. Cellular localization of these proteins was observed using immunofluorescence. Results:The expression of m 6A methylation-related proteins was gradually reduced in CVA16-infected cells with time, but showed no obvious change in ICR suckling mice or SCRBA2 humanized mice. After infection, m 6A methylation-related proteins were redistributed in both the nucleus and cytoplasm and even degraded. Conclusions:CVA16 replication in host cells altered the expression and cellular localization of m 6A methylation-related proteins, which indicated that m 6A modification might be a new potential target for enterovirus therapy.

Objective To establish a method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture by UV-Vis Spectrophotometry. Methods The contents of total flavones and polysaccharides in Fangshu Qingre mixture were determined by UV-Vis spectroscopy with rutin and anhydrous glucose as reference substance, and the wavelength was set at 508 nm and 487 nm. Results The contents were from 0.00 to 59.20 μg/ml for total flavones and from 10.92 to 109.20 μg/ml for total polysaccharides in Fangshu Qingre mixture. The recoveries of total flavones and total polysaccharides were 104.4% and 104.8% respectively. Conclusion The method of using ultraviolet spectroscopy was simple, reproducible, accurate and reliable, which could be preferably used as the method for content determination of total flavones and polysaccharides in Fangshu Qingre mixture.