Objective To investigate the combined effects of somatostatin (SST) and cisplatin (DDP) on proliferation and apoptosis in gallbladder cancer cells,and to investigate the mechanism of the combined effects.Methods We performed immunohistochemistry to detect the PTEN expression in gallbladder cancer.We then investigated the combined effects of SST and DDP on cell proliferation in vitro with CCK-8 assay and analyzed the interaction between these two drugs using isobologram analysis.We also investigated the combined effects on cell proliferation in vivo using a xenograft nude mouse model.FITC-Annexin V/PI assay and TUNEL staining assay were performed to detect the proportion of apoptosis after combined treatment in vitro and in vivo.Reactive oxygen species and mitochondrial membrane potential were detected with DCFH-DA assay and JC-1 staining assay after the combined treatment.We finally detected the PTEN and p-AKT associated proteins using western blotting after the combined treatment.Results PTEN was abnormally decreased in gallbladder cancer tissues.PTEN expression was negatively correlated with cancer differentiation and was positively correlated with patients'survival time.DDP treatment decreased while combined treatment with SST induced PTEN expression and inhibited AKT activation by reversing resistance to DDP.Isolated SST or DDP treatment inhibited gallbladder cancer GBC-SD and SGC996 cell proliferation which was dose-dependence.These two drugs synergistically inhibited gallbladder cancer cell growth in vivo and in vitro.Isolated SST or DDP treatment induced cell apoptosis and combined treatment induced cell apoptosis the most.SST inhibited AKT activation but did not induce ROS.DDP induced ROS resulting in increased cell apoptosis.Either SST or DDP alone increased the levels of cytoplasmic cytochrome C protein and activated caspase-3.Conclusions SST enhanced growth inhibition by cisplatin in gallbladder cancer cells through inducing PTEN expression.This study provides the theoretical basis for further combined clinical chemotherapeutic applications.

Objective To investigate the predictive value of preoperative serum tumor markers test for lymph node metastasis of intrahepatic cholangiocarcinoma (ICC).Methods The retrospecgtive cohort study was conducted.The clinicopathological data of 69 patients with ICC who were admitted to the Xinhua Hospital of Shanghai Jiaotong University between May 2006 and May 2016 were collected.Among 69 patients with pathological diagnosis,24 with lymph node metastasis were allocated into the lymph node metastasis group and 45 without lymph node metastasis were allocated into the non-lymph node metastasis group.Tumor markers of the 2 groups were preoperatively detected,including alpha-fetoprotein (AFP),carcinoembryonic antigen (CEA),prostate specific antigen (PSA),CA19-9,CA125,CA242,CA153,CA724,CA211,neuron-specific enolase (NSE) and squamous cell carcinoma antigen (SCC).Receiver operating characteristic (ROC) curve was built,and critical value,sensitivity and specificity were calculated based on ROC curve.Coincident rate between significant indicators and results of pathological examination was calculated.Observation indicators:(1) overall positive rates of tumor markers;(2) comparison of tmmor markers levels in the 2 groups;(3) tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination.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 (Q25,Q75) and comparison between groups was analyzed using the Wilcoxon rank sum test.Comparison of count data was analyzed by the chi-square test and Fisher exact probability.The statistically significant indicators were analyzed by the ROC curve.Results (1) Overall positive rates of tumor markers:positive rates of AFP,CEA,PSA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC in 69 patients were 27.5％ (19/69),29.0％ (20/69),4.3％ (3/69),69.6％ (48/69),36.2％ (25/69),50.7％ (35/69),26.1％ (18/69),21.7％ (15/69),62.3％ (43/69),31.9％(22/69) and 21.7％(15/69),respectively.Positive rates of AFP,CEA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC were more than 20％,which became comparison indicators of 2 groups.(2) Comparison of tumor markers levels in the 2 groups:levels of CA19-9,CA125,CA242 and CA211 were 284.9 U/mL (42.5 U/mL,730.3 U/mL),63.6 U/mL (23.4 U/mL,172.1 U/mL),71.7 U/mL (25.6 U/mL,138.9 U/mL),6.7 μg/L (3.9 μg/L,17.5 μg/L) in the lymph node metastasis group and 58.0 U/mL (25.9 U/mL,405.9 U/mL),18.2 U/mL (11.7 U/mL,33.8 U/mL),11.0 U/mL (3.7 U/mL,41.7 U/mL),3.7 μg/L (2.7 μg/L,6.9 μg/L) in the non-lymph node metastasis group,respectively,with statistically significant differences between the 2 groups (Z=2.016,3.213,3.143,2.482,P＜0.05).(3) Tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination:area under the ROC curve of CA19-9,CA125,CA242 and CA211 were respectively 0.648 [95％ confidence interval (C I):0.515-0.781,P＜0.05],0.736 (95％ CI:0.608-0.864,P＜0.05),0.731 (95％ CI:0.603-0.859,P＜0.05),0.714 (95％ CI:0.581-0.847,P＜0.05).The positive critical value,sensitivity and specificity of CA19-9,CA125,CA242 and CA21 were 150.6 U/mL,35.7 U/mL,43.4 U/mL,6.0 μg/L and 62.5％,66.7％,70.8％,62.5％ and 71.1％,82.2％,77.8％,75.6％,respectively.The coincident rate between ROC curve and results of postoperative pathological examination of CA 19-9,CA 125,CA242 and CA211 were 68.1％ (47/69),76.8％(53/69),75.4％(52/69),71.0％(49/69),respectively.Conclusion The levels of serum CA19-9,CA125,CA242 and CA211 can effectively predict lymph node metastasis of patients with ICC.

Nosocomial infections caused by gram-negative opportunistic pathogens, including Acinetobacter baumannii ( A. baumannii) and Klebsiella pneumoniae ( K. pneumoniae), pose a great challenge to health care management and human health. New treatment strategies are urgently needed to tackle with the spread of multi-drug resistant strains and the increase in bacterial virulence. Type Ⅵ secretion system (T6SS), a conservative secretory apparatus encoded in a variety of gram-negative bacteria, can inject effectors into other prokaryotic or eukaryotic cells in a contact-dependent manner to achieve antibacterial and anti-host properties. It is closely related to the environmental adaptability, competitiveness and colonization ability of bacteria in hosts. The T6SS gene cluster is composed of core genes, effector genes and associated genes, and the effectors encoded by it are highly diverse and play an important role in pathogen infection. This review summarized the advances in A. baumannii and K. pneumoniae T6SS in terms of competition, host colonization, interaction between conjugative plasmids and expression regulation, aiming to provide reference for future study on T6SS-related antimicrobial activity, virulence and resistance.

Under the circumstances of the rapid development of etiological diagnostic technology and the increasing application of new testing technologies to microbial detection, laboratory workers and clinical related departments should promptly propose Chinese standards, Chinese guidelines, and Chinese diagrams, and always adhere to the promotion and application of clinical microbiology related standards and guidelines in clinical practice, to continue to promote the virtuous cycle of standardization of etiology diagnosis, and gradually improve the laboratory diagnosis ability and technological progress of infectious diseases in China.

The 30th annual meeting of the Japanese Society of Hepato-Biliary-Pancreatic Surgery (JSHBPS) was held in Yokohama,Japan between June 7 and 9,2018.The latest research trends and advances in biliary tract cancer from this meeting were summarized and shared in this paper.The main content includes:history of biliary cancer surgery for recent 30 years and the future of HBP surgery for malignancies,establish of worldwide prospective database of biliary cancer and identify the optimal treatment for early gallbladder cancer,clinical research of neoadjuvant / adjuvant therapy,and progress in basic research of biliary malignancies.

To investigate the resistance mechanisms of tigecycline-non-susceptible Acinetobacter baumannii and for providing the evidence of the control of nosocomial infection and rational use of antibiotics. The minimum inhibitory concentrations (MICs) of 94 non repetitive tigecycline-non-susceptible A. baumannii from 20 hospitals in 12 cities of China were determined by agar dilution method and broth microdilution method. The molecular epidemiology was studied by Multilocus sequence typing (MLST) and eBURST software. PCR and sequencing techniques were used to analyze the resistance genes (blaOXA-40-like, blaOXA-58-like, blaOXA-23-like, blaOXA-51-like, blaNDM-1), ISAba1, and the mutation sites of adeR, adeS, and trm. The activity of polymyxin B and minocyclinem against tigecycline-non-susceptible A. baumannii were 100% and 25.5%, respectively. The sensitivities of other antibiotics were less than 3.5%, and the sensitivities of imipenem and meropenem totigecycline-nonsusceptible A. baumannii were only 1.1%. A total of 12 ST types were identified, including ST195 (45, 47.9%), ST208 (19, 20.2%) and ST457 (10, 10.6%). EBURST analysis found that 8 of the ST types belonged to the clone complex 92 (Clonal Complex 92, CC92). The blaOXA-23-like type carbapenem gene was identiefied in 93 strains (99% positive); and none of the strains contained the blaNDM-1 gene. The detection rates of adeR and adeS were 73.4% and 91.5% respectively and high frequency mutation sites were located in adeR (Asp26Asn) and adeS (Ala97Glu); The ISAba1 located upstream of the adeS gene was detected in 12 strains of A. baumannii, mainly from the northern region of China. The 240 nucleotide deletion of the trm gene caused a frameshift leading to a premature stop. So the tigecycline-non-susceptible A. baumannii showed high resistance against most antibiotics except polymyxin B. The deletion and mutation of adeR, adeS and trm were the main resistant mechanisms in tigecycline-non-susceptible A. baumannii in China.

To dynamically investigate the distribution and antimicrobial resistance profiles of bacteremia pathogens isolated from different regions in China in 2011, 2013 and 2016. Non-repetitive isolates from nosocomial bloodstream infections were retrospectively collected and detected for antimicrobial susceptibility tests (AST) by agar dilution or microbroth dilution methods. Whonet 5.6 was used to analyze the AST data. Among 2 248 isolates, 1 657 (73.7%) were Gram-negative bacilli and 591 (26.3%) were Gram-positive cocci. The top five bacteremia pathogens were as follows, Escherichia coli (32.6%, 733/2 248), Klebsiella pneumoniae (14.5%, 327/2 248), Staphylococcus aureus (10.0%, 225/2 248), Acinetobacter baumannii (8.7%, 196/2 248) and Pseudomonas aeruginosa (6.2%, 140/2 248). Colistin (96.5%, 1 525/1 581, excluding innate resistant organisms), tigecycline (95.6%, 1 375/1 438, excluding innate resistant organisms), ceftazidine/clavulanate acid (89.2%, 1 112 /1 246), amikacin (86.4%, 1 382/1 599) and meropenem (85.7%, 1 376/1 605) showed relatively high susceptibility against Gram-negative bacilli. While tigecycline, teicoplanin and daptomycin (the susceptibility rates were 100.0%), vancomycin and linezolid (the susceptibility rates were 99.7%) demonstrated high susceptibility against Gram-positive cocci. The prevalence of extended-spectrum β-lactamases (ESBLs)-producing Enterobacteriaceae were 50.6% (206/407), 49.8% (136/273) and 38.9% (167/429) in 2011, 2013 and 2016 respectively; carbapenem-non-susceptible Enterobacteriaceae were 2.2% (9/408), 4.0% (16/402) and 3.9% (17/439) in 2011, 2013 and 2016 respectively; The prevalence of multidrug-resistant A. baumannii (MDRA) was 76.4% (55/72) in 2011, 82.7% (43/52) in 2013 and 87.5% (63/72) in 2016, respectively. The prevalence of multidrug-resistant P. aeruginosa (MDRP) was 9.8% (5/51) in 2011, 20.0% (7/35) in 2013 and 13.0% (7/54) in 2016, respectively. The prevalence of methicillin-resistant S. aureus (MRSA) was 51.9% (41/79) in 2011, 29.7% (19/64) in 2013 and 31.7% (26/82) in 2016, respectively. The prevalence of high level gentamicin resistance (HLGR) of Enterococcus faecium and Enterococcus faecalis were 43.2% (48/111) and 40.9% (27/66), respectively. The predominant organism of carbapenem-non-susceptible Enterobacteriaceae was K. pneumoniae with its proportion of 57.1% (24/42). Among 30 tigecycline-non-susceptible Enterobacteriaceae, K. pneumoniae was the most popular organism with 76.7% (23/30). Among 39 colistin-resistant Enterobacteriaceae, E. coli, Enterobacter cloacae and K. pneumoniae were constituted with the percent of 43.6 (17/39), 35.9 (14/39) and 15.4 (6/39), respectively. The Gram-negative bacilli (E. coli and K. pneumoniae were the major organisms) were the major pathogens of nosocomial bacteremia, to which tigecycline, colistin and carbapenems kept with highly in vitro susceptibility. Whereas, among the Gram-positive cocci, S. aureus was the top 1 isolated organism, followed by E. faecium, to which tigecycline, daptomycin, linezolid, vancomycin and teicoplanin kept with highly in vitro susceptibility. Isolation of colistin-resistant Enterobacteriaceae, tigecycline-non-susceptible Enterobacteriaceae, linezolid- or vancomycin-non-susceptible Gram-positive cocci suggests more attention should be paid to these resistant organisms and dynamic surveillance was essential.