Objective To understand the distribution and drug resistance of pathogens from patients with anastomotic fistula infection after esophageal cancer surgery, and provide basis for clinical diagnosis and treatment.Methods Patients were admitted to a hospital due to anastomotic fistula after esophageal cancer surgery between January 2012 and December 2015, microbial culture and antimicrobial susceptibility testing results of patients were retrospectively analyzed.Results 1 986 patients underwent radical resection of esophageal cancer within 4 years, 148 of whom developed anastomotic fistula, 104 (70.27％) were with positive microbial culture.A total of 197 pathogenic strains were isolated, 52(26.40％)and 145 (73.60％)strains were isolated from intrathoracic anastomotic fistula and cervical anastomotic fistula respectively;127 (64.47％)strains were gram-negative bacteria, the major were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii, 62(31.47％) strains were gram-positive bacteria, the major were Staphylococcus aureus, Enterococcus spp., and Streptococcus viridans, 8 strains (4.06％) were fungi.49(47.12％) cases were with mixed pathogenic infection.The resistance rates of gram-negative bacteria to imipenem were 17.86％-47.62％, to polymyxin B was 0, resistance rates of Pseudomonas aeruginosa to other antimicrobial agents were all<50％, Klebsiella pneumoniae to piperacillin and aztreonam were both>70％, Acinetobacter baumannii to most antimicrobial agents were all>50.00％;resistance rates of gram-positive bacteria to clindamycin and tetracycline were both>50.00％, to linezolid, vancomycin, and teicoplanin were all 0, resistance rates of Staphylococcus aureus to penicillin, oxacillin, and ciprofloxacin were all>60％,resistance rate of Enterococcus spp.to quinupristin/dalfopristin was 100.00％.Conclusion Postoperative anastomotic fistula combined with infection can affect the prognosis of patients after esophageal cancer surgery, regular monitoring on distribution and drug resistance of pathogens can provide the basis for initial empirical treatment, and is conducive to the early treatment and rational use of antimicrobial agents.

Objective To establish and apply the procedure of survey on quality indicator in clinical laboratory and to analyze the status in quo of the 15 quality indicators in Zhejiang province .Methods A network platform for the survey on quality indicator in clinical laboratory was designed and developed by our center.The online questionnaires that should be reported back within one month were assigned to 473 laboratories.The developed software and SPSS 13.0 were used for statistical analysis .13 indicators expressed in rate were further evaluated with sigma scales .The 25th percentile, 50th percentile, and 75th percentile of the distribution of each quality indicator were regarded as the minimum , appropriate and optimum quality specifications, respectively.Results Totally 444 laboratories submitted the survey results.The overall sigma levels of 10/13 indicators were all >3, of which the inappropriate CV of internal quality control and unacceptable performances in EQA were still less than 3σin 15.8%and 9.2%of the laboratories.The rates of quality indicators in different scales of laboratories and diverse disciplines were significantly different .Pre-analytical TAT in routine examination for clinical chemistry and immunology was 50 min, on average.And the time for routine examination of blood , urine and stool was 30 min.Pre-analytical TAT in emergency examination for all four disciplines were all between 10 and 15 min. Intra-analytical TAT for clinical immunology was the longest , which was 154 min for routine examination and 40 min for emergency examination, respectively.The optimum quality specifications for 8 indicators were 6σ, while the minimum quality specifications were less than 1σfor 4 indicators.Conclusions According to the results of our survey, the pre-analytical quality indicator perform better than that of Intra-analytical and post-analytical phase.The laboratory should strengthen the laboratory information system technology construction to ensure the reliable data collection and long-time monitoring.

OBJECTIVETo identify products decomposed in high temperature water and alcohol from diester alkaloids such as aconitum alkaloid, in order to study their transformation regularity.

METHODStructures of multiple converted products were determined by analyzing on multistage mass spectrometry of known compounds and literature searching.

RESULTBenzaconine and pyraconitine were the major hydrolysates, while pyraconitine and ethoxy-aconitine were the major alcoholysates from diester alkaloids.

CONCLUSIONPyraconitine alkaloids, as pyrolytic products, are not related to the type of solvent. 8-ethoxy-aconitine alkaloids, as alcoholysates, are related to the type of solvent. This study identifies multiple converted products from alkaloids and summarizes mass spectrometry fragmentation regularity by LC-MS, laying a firm foundation for studies on the transformation of toxic diester alkaloids contained in aconitum and providing a basis for studies on the transformation of alkaloids contained in aconitum during boiling.

Aconitum ; chemistry ; Alkaloids ; chemistry ; Ethanol ; chemistry ; Hydrolysis ; Mass Spectrometry

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 effect and mechanism of miR-195 regulating FOXK1 gene and PI3K/Akt pathway on stomach adenocarcinoma proliferation, invasion and migration ability.Methods:Public database samples were employed to analyze the expression differences and prognostic significance of miR-195 in stomach adenocarcinoma. After overexpression of mir-195-5p in two cell lines, MGC803 and AGS, altered cell proliferation, invasion, and migration abilities were detected by Alamar Blue, Wound healing, and Transwell assays. The potential target genes and binding sites of miR-195 were predicted by the starBase. Western blot was used to detect the expression levels of foxk1 and phosphorylation sites in the PI3K/Akt pathway of target genes after overexpression of mir-195-5p. A Dual-luciferase reporter assay was used to verify the relationship between mir-195-5p and foxk1. Statistical analyses were performed with IBM SPSS 22 software and R 4.0.3.Results:Our results showed a significant over-expression of miR-195 in the tumor tissues, compared with the paired normal tissues ( P<0.001) , which could inhibit the proliferation and invasion of stomach carcinoma cells and significantly correlated with survival ( P=0.011) . Moreover, our study indicated that miR-195 depressed the expression of FOXK1 and significantly reduced the activation of the PI3K/Akt pathway, which had a negative effect on the proliferation and invasion of stomach carcinoma cells. The phosphorylated Akt (s473 site) expression in the PI3K/Akt pathway was significantly decreased after overexpression of miR-195. Conclusion:Overall, our studies clarify the important function of the miR-195 in the diagnosis and therapy of patients with stomach carcinoma and reveal the FOXK1 and PI3K/Akt pathway regulation by the miR-195, which are of important clinical significance in the differential diagnosis.

Uch37 is a de-ubiquitinating enzyme that is activated by Rpn13 and involved in the proteasomal degradation of proteins. The full-length Uch37 was shown to exhibit low iso-peptidase activity and is thought to be auto-inhibited. Structural comparisons revealed that within a homo-dimer of Uch37, each of the catalytic domains was blocking the other's ubiquitin (Ub)-binding site. This blockage likely prevented Ub from entering the active site of Uch37 and might form the basis of auto-inhibition. To understand the mode of auto-inhibition clearly and shed light on the activation mechanism of Uch37 by Rpn13, we investigated the Uch37-Rpn13 complex using a combination of mutagenesis, biochemical, NMR, and small-angle X-ray scattering (SAXS) techniques. Our results also proved that Uch37 oligomerized in solution and had very low activity against the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) of de-ubiquitinating enzymes. Uch37Δ(Hb,Hc,KEKE), a truncation removal of the C-terminal extension region (residues 256-329) converted oligomeric Uch37 into a monomeric form that exhibited iso-peptidase activity comparable to that of a truncation-containing the Uch37 catalytic domain only. We also demonstrated that Rpn13C (Rpn13 residues 270-407) could disrupt the oligomerization of Uch37 by sequestering Uch37 and forming a Uch37-Rpn13 complex. Uch37 was activated in such a complex, exhibiting 12-fold-higher activity than Uch37 alone. Time-resolved SAXS (TR-SAXS) and FRET experiments supported the proposed mode of auto-inhibition and the activation mechanism of Uch37 by Rpn13. Rpn13 activated Uch37 by forming a 1:1 stoichiometric complex in which the active site of Uch37 was accessible to Ub.
Binding Sites ; Catalytic Domain ; Chromatography, Gel ; Crystallography, X-Ray ; Humans ; Membrane Glycoproteins ; chemistry ; genetics ; metabolism ; Nuclear Magnetic Resonance, Biomolecular ; Protein Binding ; Protein Conformation ; Protein Multimerization ; Scattering, Small Angle ; Ubiquitin Thiolesterase ; chemistry ; genetics ; metabolism ; Ultracentrifugation