AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.

Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P

Objective:To understand the current situation of effective teaching behavior of clinical nursing teachers and analyze it.Methods:A convenient sampling method was used to conduct a questionnaire survey on nursing students and teaching teachers in Tai′an City central hospital and the Second Affiliated Hospital of Shandong First Medical University, using Cui Shasha Chinese version of Nursing Clinical Teacher Effectiveness Inventory as the research tool.Results:The score of the teachers in the "interpersonal relationship" dimension was the highest (6.32±0.89), while the score in the "nursing ability" dimension was the lowest (5.95±0.99). There was no statistically significant difference between college students and undergraduate students in the evaluation of effective teaching behaviors of teachers, while there was statistically significant difference between nursing students (including college students and undergraduate students) and teachers in all dimensions and total scores of the scale ( t values were -4.616--2.250, P<0.05 or 0.01). Teachers and students were more consistent with the 10 items with low scores than the 10 items with high scores (the total number of items was 2 and 5, respectively). Conclusions:The teaching teacher should be able to correctly identify and judge the learning needs of the internship nursing students, so that the teaching effect meets the expectations of the internship nursing students, and should be targeted when teaching internship nursing students of different educational levels as well as focusing on nursing research.

Objective:To systematically evaluate the effects of different umbilical cord time on maternal and fetal outcomes.Methods:Meta analysis was used, CNKI, Wanfang, VIP, PubMed, The Cochrane Library, EBSCO, Web of Science database, etc. were searched. The literatures were screened according to the inclusion and exclusion criteria, and the literature quality was evaluated. Revman 5.2 software was used for data processing and analysis.Results:A total of 9 randomized controlled trials were included. Meta-analysis showed that delayed umbilical cord did not increase postpartum 2,24 h hemorrhage[mean difference ( MD)=-40.69, 95%confidence interval (CI) (-83.05-1.67), P>0.05 and MD=-10.33, 95% CI (-22.47-1.81), P>0.05] and did not prolong the third stage of labor [ MD= 0.32, 95% CI (-0.71-1.35), P>0.05] compared with conventional umbilical cord (early umbilical cord) method. 1,5 min Apgar scores for different neonatal umbilical cord time was not statistically significant[ MD=0.05, 95% CI (-0.02-0.12), P>0.05 and MD=-0.01, 95% CI (-0.04-0.03), P>0.05]. There was no significant difference in neonatal weight [ MD=72.91, 95% CI (-53.98-199.80), P>0.05]. Delayed umbilical cord could improve the newborn's 24 h capillary hemoglobin [ MD=17.69, 95% CI (13.55-21.83), P<0.01] and bilirubin peak [ MD=17.50, 95% CI (11.88-23.12), P<0.01]. Conclusions:Delayed umbilical cord and early umbilical cord did not increase postpartum hemorrhage and prolong the third stage of labor, and had no adverse effects on maternal. The peak levels of capillary hemoglobin and bilirubin in newborns were increased 24 h, but there was no significant difference in neonatal 1 min and 5 min Apgar scores and neonatal weight.

Objective To explore the mechanism of mesenteric lymph duct ligation relieving hepatic injury in rats by two-hit of hemorrhage and LPS.Methods Forty-five Wistar rats were divided into three groups: ligation group,non-ligation group and sham group,and the two-hit model was established by hemorrhage and LPS,mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group.After 24 hours of operation,took out the liver for pathological section,and the hepatocellular apoptosis rate was determined by method of TUNEL,the expression of BCL-2 and BAX protein was determined by immunohistochemical test.At the same time,taking out liver for homogenate of 10 percent,the activity of MPO and ATPase and the contents of TNF-? and IL-6 were determined in hepatic homogenate.Results After two-hit,the hepatocellular apoptosis rate and expression of BAX protein in non-ligation group were significantly increased as compared with sham group and ligation group,and expression of BCL-2 protein was significantly lower.The contents of MPO,TNF-? and IL-6 in hepatic homogenate of non-ligation group were significantly increased than that of sham group,and the activity of ATPase in hepatic homogenate was significantly lower.But the ATPase in hepatic homogenate of ligation group were significantlyincreased and MPO,TNF-? and IL-6 in hepatic homogenate of ligation group were significantly lower as compared with non-ligation group.Conclusion The mechanism of mesenteric lymph duct ligation relieving hepatic injury of rats was related to the mesenteric lymph blockage reduces the TNF-? and IL-6 and improves the expression of BCL-2 protein and the activity of ATPase in liver.

Objective To investigate the changes of serum Copeptin and matrix metalloproteinase-9 (MMP-9) in children with chronic heart failure (CHF) and its clinical significance. Methods A total of 186 children with CHF were selected for CHF group, including 78 cases of cardiac function grade Ⅱ, 65 cases of grade Ⅲ, and 43 cases of grade Ⅳ. There were 57 cases of dilated cardiomyopathy, 68 cases of congenital heart disease and 61 cases of other diseases. Another 85 healthy children from health checkup were chosen as controls. The levels of serum Copeptin and MMP-9 were determined by enzyme linked immunosorbent assay (ELISA), and the level of N-terminal pro-brain natriuretic peptide (NT-proBNP) was measured by bidirectional lateral flow immunoassay. The left ventricular end diastolic dimension (LVEDD), left ventricular end systolic dimension (LVESD), left ventricular ejection fraction (LVEF), and left ventricular short fraction shortening (LVFS) were measured by echocardiography. ROC curve was used to analyze the diagnostic value of serum Copeptin and MMP-9 in CHF. The correlation of serum Copeptin and MMP-9 with the cardiac function indices were examined by Pearson correlation analysis. Results The levels of serum copeptin, MMP-9, and NT-proBNP in different cardiac function groups (Ⅱ, Ⅲ, Ⅳ) increased gradually with the aggravation of the cardiac function damage and were higher than those in control group, and the differences were statistically significant (P<0.05). Compared with the control group and cardiac function grade Ⅱ group, the levels of LVESD and LVEDD were increased and the levels of LVEF and LVFS were decreased in the grade Ⅲ and Ⅳ groups. Compared with the grade Ⅲ group, the levels of LVESD and LVEDD were increased and the levels of LVEF and LVFS were decreased in the grade Ⅳ groups. There were significant differences (P<0.05). The ROC curve showed that the area under the curve (AUC) and 95％ CI of serum Copeptin, MMP-9, NT-proBNP and combinations of these three biomarkers in the diagnosis of CHF were 0.845 (0.781～0.914), 0.806 (0.736～0.883), 0.894 (0.828～0.962) and 0.925 (0.846～0.983) respectively, and the optimal thresholds were 12.5 pmol/L, 175.3 μg/L and 2037.0ng/L. The level of serum Copeptin was positively correlated with MMP-9 (r=0.807, P<0.001). Conclusion Serum Copeptin and MMP-9 may be involved in the ventricular remodeling in CHF children and they are expected to be a good indicator for the diagnosis of CHF and cardiac function.

Peroral endoscopic myotomy (POEM) has emerged as a rescue treatment for recurrent or persistent achalasia after failed initial management. Therefore, we aimed to investigate the efficacy and safety of POEM in achalasia patients with failed previous intervention. We searched the MEDLINE, Embase, Cochrane, and PubMed databases using the queries “achalasia,” “peroral endoscopic myotomy,” and related terms in March 2019. Data on technical and clinical success, adverse events, Eckardt score and lower esophageal sphincter (LES) pressure were collected.The pooled event rates, mean differences (MDs) and risk ratios (RR) were calculated. A total of 15 studies with 2,276 achalasia patients were included. Overall, the pooled technical success, clinical success and adverse events rate of rescue POEM were 98.0% (95% confidence interval [CI], 96.6% to 98.8%), 90.8% (95% CI, 88.8% to 92.4%) and 10.3% (95% CI, 6.6% to 15.8%), respectively. Seven studies compared the clinical outcomes of POEM between previous failed treatment and the treatment naïve patients. The RR for technical success, clinical success, and adverse events were 1.00 (95% CI, 0.98 to 1.01), 0.98 (95% CI, 0.92 to 1.04), and 1.17 (95% CI, 0.78 to 1.76), respectively. Overall, there was significant reduction in the pre- and post-Eckardt score (MD, 5.77; p<0.001) and LES pressure (MD, 18.3 mm Hg; p<0.001) for achalasia patients with failed previous intervention after POEM. POEM appears to be a safe, effective and feasible treatment for individuals who have undergone previous failed intervention. It has similar outcomes in previously treated and treatment-naïve achalasia patients.

Objective:To discuss the necessity and feasibility of constructing the baccalaureate midwifery education standard in China by the comparative study of WHO and ICM education standards, combined with China's medical nursing education standards.Methods:We searched the scientific literature using the databases of CNKI, Wanfang, VIP and the official websites of WHO and ICM. Comparative study was conducted combined with the literature study, analyze the specific fields and indicators required for baccalaureate midwifery education standards in China.Results:ICM standard is more accord with the actual needs of the construction of baccalaureate midwifery education standard in China.Conclusion:It is feasible and necessary to construct the baccalaureate midwifery education standard according to ICM standards, this study further provides reference and basis for constructing the baccalaureate midwifery education standard in China.

Objective To observe the value of quantitative CT(QCT)abdominal body fat parameters combined with clinical-imaging model for predicting recurrent acute pancreatitis(RAP).Methods Data of 468 patients with acute pancreatitis(AP),including 207 cases of RAP(RAP group)and 261 cases of non-RAP(non-RAP group)were retrospectively analyzed.Clinical information,conventional CT manifestations and QCT parameters such as abdominal subcutaneous fat area(SFA),visceral fat area(VFA),total abdominal fat area(TFA),hepatic fat content and pancreatic fat content at the first visit were recorded or measured.Clinical characteristics,CT manifestations and QCT parameter values were compared between groups,and the independent factors for predicting RAP were selected with multivariate logistic regression analysis.Then a clinical-imaging model and a comprehensive model combining with QCT parameters were constructed,and their efficacies for predicting RAP were evaluated with receiver operating characteristic(ROC)curves,and the area under the curves(AUC)were calculated.Results Compared with non-RAP group,patients in RAP group were younger,had higher percentage of alcohol consumption,biliary stones and hyperlipidemia,as well as of distinct pancreatic margins on CT images,also higher VFA,TFA,liver fat content and pancreatic fat content(all P＜0.05).Alcohol consumption,hyperlipidemia,biliary stones,pancreatic margins and pancreatic fat content were all independent predictors of RAP,and the comprehensive model constructed based on these five had higher AUC(0.860)than clinical-imaging model constructed based on the previous 4 factors(0.701)and pancreatic fat content alone(0.770)(both P＜0.001).Conclusion QCT abdominal body fat parameters combined with clinical-imaging comprehensive model was effective for predicting the risk of RAP.

Macrolide antibiotics are a class of broad-spectrum antibiotics with the macrolide as core nucleus. Recently, antibiotic pollution has become an important environmental problem due to the irregular production and abuse of macrolide antibiotics. Microbial degradation is one of the most effective methods to deal with antibiotic pollution. This review summarizes the current status of environmental pollution caused by macrolide antibiotics, the degradation strains, the degradation enzymes, the degradation pathways and the microbial processes for degrading macrolide antibiotics. Moreover, the critical challenges on the biodegradation of macrolide antibiotics were also discussed.
Anti-Bacterial Agents ; Biodegradation, Environmental ; Macrolides