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Objective:To investigate the role of interleukin-13(IL-13), interleukin-13 receptor α2(IL-13Rα2) and 11 β -hydroxysteroid dehydrogenase 2(11βHSD2) signaling pathway in liver metastasis of colon cancer and its mechanism.Methods:A retrospective case-control study was conducted to analyze the clinical data of 80 patients with colorectal cancer who were operated in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 2015 to December 2018.All patients were followed up by clinic or telephone until August 30, 2019.According to the occurrence of liver metastasis, the patients were divided into metastasis group ( n=22) and non metastasis group ( n=58). Real-time fluorescence quantitative Polymerase Chain Reaction(PCR) and Western blotting were used to detect and compare the mRNA relative expression and protein expression of IL-13, IL-13Rα2, 11βHSD2, cyclooxygenase 2(COX 2) and protein kinase B in cancer tissues and cancer adjacent tissues. Glycyrrhetinic acid, an inhibitor of 11 β HSD2, was used to inhibit the activity of 11hsd2 in human colon cancer cell line HCT-8.The mRNA relative expression and protein expression of IL-13, IL-13Rα2, 11βHSD2, COX 2 and protein kinase B were detected by real-time fluorescence quantitative PCR and Western blotting before and 24 hours after glycyrrhetinic acid addition. Results:The mRNA relative expression of IL-13(0.79±0.11, 0.40±0.10), IL-13Rα2(0.72±0.13, 0.46±0.11), 11βHSD2(0.84±0.26, 0.60±0.08), COX 2(0.70±0.25, 0.37±0.04), protein kinase B (0.76±0.13, 0.42±0.06) in colon cancer tissues of metastatic and non metastatic groups were higher than those in cancer adjacent tissues(0.09±0.01, 0.10±0.06, 0.09±0.02, 0.09±0.03, 0.09±0.01, 0.09±0.02, 0.13±0.02, 0.12±0.07, 0.05±0.02, 0.05±0.03). The difference was statistically significant (t value was 28.36, 23.20, 22.07, 24.88, 16.47, 16.47, 47.86, 18.55, 24.55, 26.20, 44.40, all P<0.001). The protein expression of IL-13(0.48±0.11, 0.32±0.07), IL-13Rα2(0.52±0.11, 0.36±0.11), 11βHSD2(0.63±0.12, 0.48±0.11), COX2(0.45±0.15, 0.27±0.09), protein kinase B(0.50±0.12, 0.29±0.08) in colon cancer tissues of metastatic and non metastatic groups were higher than those in cancer adjacent tissues(0.12±0.02, 0.13±0.01, 0.10±0.02, 0.10±0.02, 0.14±0.06, 0.13±0.05, 0.10±0.03, 0.10±0.04, 0.10±0.03, 0.10±0.02). The difference was statistically significant ( t value were 15.63, 21.15, 17.71, 17.28, 11.01, 18.14, 10.55, 13.12, 15.76 and 18.90 respectively, all P<0.001). The relative expression of mRNA and protein in metastasis group was higher than that in non metastasis group ( t=15.15, 3.01, 8.97, 2.52, 6.34, 2.26, 9.82, 2.52, 16.02, 3.57, respectively, all P< 0.05). Compared with that those of before glycyrrhetinic acid addition, after 24 hours of glycyrrhetinic acid addition, the mRNA relative expression and protein expression of IL-13, IL-13Rα2 had no significant change (all P>0.05), while the mRNA relative expression and protein expression of COX 2 and protein kinase B(before adding: 0.725±0.159, 0.639±0.162, 0.741±0.178, 0.668±0.145, after adding: 0.108±0.085, 0.116±0.048, 0.122±0.063, 0.119±0.066) were decreased( t value were 18.744, 16.954, 17.956, 18.875 respectively , all P<0.01). Conclusion:The activation of IL-13/IL-13α2/11βHSD2 signaling pathway can promote liver metastasis of colon cancer.The mechanism may be that 11βHSD2 over expression promotes cancer cells invasion and migration-related COX2 expression and PI3K/protein kinase B pathway, and promotes liver metastasis of colon cancer.
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Objective:To investigate the effects of different lymph node dissection methods in radical resection of intrahepatic cholangiocarcinoma (ICC) on the number of harvested lymph nodes and prognosis of patients.Methods:The retrospective cohort study was conducted. The clinicopathological data of 57 patients with ICC who underwent radical resection in the Second Affiliated Hospital of Nanchang University from January 2011 to January 2016 were collected. There were 25 males and 32 females, aged from 45 to 72 years, with an average age of 59 years. Of the 57 patients, 36 undergoing en bloc fusion lymph node dissection and 21 undergoing conventional lymph node dissection were allocated into observation group and control group, respectively. Observation indicators: (1) intraoperative situations; (2) results of postoperative pathological examination; (3) postoperative complications; (4) follow-up. Follow-up using outpatient examination or telephone interview was performed once every six months after hospital discharge to detect survival of patients up to January 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was done using the t test. Measurement data with skewed distribution were described as M (range), and comparison between groups was analyzed by the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact propability. Comparison of ordinal data between groups was analyzed using the Mann-Whitney U test. The survival curve and rate were drawn and calculated using the Kaplan-Meier method, and survival of patients was analyzed by the Log-rank test. Results:(1) Intraoperative situations: operation time, volume of intraoperative blood loss, hepatic portal occlusion time, cases with <3 segments of hepatectomy and ≥3 segments of hepatectomy for the observation group were (218±53)minutes, (266±24)mL, (21±9)minutes, 5, 31, respectively, versus (207±45)minutes, (270±23)mL, (19±8)minutes, 2, 19 for the control group, showing no significant difference in the above indicators between the two groups ( t=-0.530, 0.774, 0.590, χ2=0.004, P>0.05). (2) Results of postoperative pathological examination: the number of harvested lymph nodes and percentage of positive lymph nodes of the observation group were 11±3 and 16.58%(66/398), respectively, versus 5±2 and 6.80%(7/103) of the control group, showing significant differences in the above indicators between the two groups ( t=-9.454, 2.148, P<0.05). (3) Postoperative complications: cases with Clavien-Dindo grade Ⅰ-Ⅱ complications or Clavien-Dindo grade Ⅲ-Ⅳ complications, cases with bile leakage, cases with postoperative hemorrhage, cases with incision infection, cases with gastroparesis were respectively 35, 1, 6, 1, 8, 0 in the observation group and 19, 2, 3, 1, 1, 2 in the control group. There was no significant difference in the cases with complications of Clavien-Dindo classification, cases with bile leakage, cases with incision infection between the two groups ( χ2=0.236, 0.000, 1.870, P>0.05) and no significant difference in the cases with postoperative hemorrhage, cases with gastroparesis between the two groups ( P>0.05). (4) Follow-up: 57 patients were followed up for 5-42 months, with a median time of 36 months. The survival time and postoperative 3-year survival rate were respectively 36 months (range, 8-42 months) and 66.7% of the observation group, versus 23 months (range, 5-39 months) and 38.1% of the control group, showing significant differences in the above indicators between the two groups ( Z=-2.253, χ2=5.317, P<0.05). Conclusion:For radical resection of ICC, the en bloc fusion lymph node dissection is beneficial to increase the number of harvested lymph nodes and improve survival rate of patients.
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Identifying antimicrobial resistant (AMR) bacteria in metagenomics samples is essential for public health and food safety. Next-generation sequencing (NGS) technology has provided a powerful tool in identifying the genetic variation and constructing the correlations between genotype and phenotype in humans and other species. However, for complex bacterial samples, there lacks a powerful bioinformatic tool to identify genetic polymorphisms or copy number variations (CNVs) for given genes. Here we provide a Bayesian framework for genotype estimation for mixtures of multiple bacteria, named as Genetic Polymorphisms Assignments (GPA). Simulation results showed that GPA has reduced the false discovery rate (FDR) and mean absolute error (MAE) in CNV and single nucleotide variant (SNV) identification. This framework was validated by whole-genome sequencing and Pool-seq data from Klebsiella pneumoniae with multiple bacteria mixture models, and showed the high accuracy in the allele fraction detections of CNVs and SNVs in AMR genes between two populations. The quantitative study on the changes of AMR genes fraction between two samples showed a good consistency with the AMR pattern observed in the individual strains. Also, the framework together with the genome annotation and population comparison tools has been integrated into an application, which could provide a complete solution for AMR gene identification and quantification in unculturable clinical samples. The GPA package is available at https://github.com/IID-DTH/GPA-package.
Subject(s)
Humans , Bacteria , Genetics , Bayes Theorem , DNA Copy Number Variations , Genome, Bacterial , Genotyping Techniques , High-Throughput Nucleotide Sequencing , Klebsiella pneumoniae , Genetics , Metagenomics , Methods , Polymorphism, Genetic , SoftwareABSTRACT
Two decades have passed since the first bacterial whole-genome sequencing, which provides new opportunity for microbial genome. Consequently, considerable genetic diversity encoded by bacterial genomes and among the strains in the same species has been revealed. In recent years, genome sequencing techniques and bioinformatics have developed rapidly, which has resulted in transformation and expedited the application of strategy and methodology for bacterial genome comparison used in dissection of infectious disease epidemics. Bacterial whole-genome sequencing and bioinformatic computing allow genotyping to satisfy the requirements of epidemiological study in disease control. In this review, we outline the significance and summarize the roles of bacterial genome sequencing in the context of bacterial disease control and prevention.We discuss the applications of bacterial genome sequencing in outbreak detection, source tracing, transmission mode discovery, and new epidemic clone identification. Wide applications of genome sequencing and data sharing in infectious disease surveillance networks will considerably promote outbreak detection and early warning to prevent the dissemination of bacterial diseases.
Subject(s)
Humans , Bacteria , Genetics , Bacterial Infections , Epidemiology , Microbiology , Bacterial Typing Techniques , Disease Outbreaks , Genome, Bacterial , Genotype , Population Surveillance , Whole Genome SequencingABSTRACT
Objective To analyze the characteristics of toxin, the PCR-ribotyping(RT) and the multilocus sequence typing(MLST) of Clostridium difficile strains isolated from China-Japan Friendship Hospital in order to provide a basis for monitoring the outbreak of nosocomial Clostridium difficile infection.Methods A total of 321 samples were collected from the patients with suspected Clostridium difficile infection(CDI) in China-Japan Friendship Hospital(CJFH) during 2012 to 2013.All Clostridium difficile strains were isolated and identified by the standard phenotypic culture method.Cytotoxicity test was performed to detect toxin B.Toxin genes (tcdA and tcdB) and binary toxin genes (cdtA and cdtB) harbored by those strains were analyzed.RT and MLST were used for homologous analysis.Clinical data of the patients were collected to analyze the isolation rate of Clostridium difficile in different populations.Results Forty-eight strains of Clostridium difficile were isolated from 46 patients with diarrhea and three of them were isolated from the same patient.The incidence of CDI among all patients, outpatients and inpatients were 14.3%(46/321), 12.8%(5/39) and 14.5%(41/282), respectively.Toxin B was detected in all of the strains as indicated by the cytotoxicity test.Strains of sequence type 1(ST1) showed the strongest cytotoxicity of all the isolated Clostridium difficile strains.Ten out of the 48 strains (20.8%) were tcdA(-)/tcdB(+) strains, which belonged to either ST37 or ST81.The results of RT and MLST were consistent in assigning the strains into nine types, in which the predominant type was ST1/RT027 accounting for 27.1% (13/48).All of the ST1/RT027 strains presented a toxin gene profile of tcdA(+)/tcdB(+) and cdtA(+)/cdtB(+).Most of the ST1/RT027 strains were isolated from the Traditional Chinese Medicine Department of Respiratory, where smallnosocomial outbreaks of ST1/RT027 strain infection might happen.Conclusion CDI diagnosed in CJFH mainly belongs to nosocomial infection.Most of the isolated strains harbor tcdA(+)/tcdB(+) genes.Surveillance for the outbreaks of CDI caused by ST1/RT027 strains over producing toxins A and B should be strengthened in hospitals.
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Objective Farmer' s markets with live birds (FMLB) are key sites where human infections by influenza A virus subtype H7N9 happened.Approximately 80％ cases have exposed to FMLB.This study is to investigate the geographic relationship between FMLB and human cases based on analysis of internet data of their geographic locations.Methods Using big data from internet,we searched all FMLB in the cities where the human cases have been reported,then analyzed geographic relations,and evaluated the possibility of visits of the patients to the FMLB around them.The densities of FMLB,population and live poultries were also analyzed.Results Forty-two cities and 10 615 markets were included in the study.It is indicated that the number of human cases has positive correlations with the population density,the number and density of markets.Except three markets in Foshan,human cases have been reported within 5 km of 10 of 13 markets,which shows that the live bird trading is highly relevant with the distribution of cases.We identified 13 hot spots in the cities including Hangzhou,Shenzhen,et al,where clustered cases have emerged.Conclusion The numbers of human cases are significantly high in cities where FMLB are detected positive for H7N9 virus.These virus positive markets usually affect the people's residence within 5km area.The number and location of FMLB in cities should be re-evaluated and re-planed for healthy city where the risk of residents infecting avian influenza virus is greatly reduced or eliminated.