OBJECTIVETo explore the NEK-6 expression in gastric cancer tissue and its relationship with clinicopathological features.

METHODSFluorescent quantification PCR and Western blotting were used to examine the NEK-6 expression in 36 samples of fresh gastric cancer tissues and para-cancer gastric mucosal tissues, human gastric cancer cell lines(BGC-823, MKN-28, SGC-7901, MGC-803, HGC-27, AGS), and human normal gastric epithelial cell line (GES-1). Gastric cancer cell lines with the highest expression level were selected to perform the invasion and migration tests, and the effect of down-regulated NEK-6 expression by siRNA transfection on above invasion and migration tests were observed. Meanwhile NEK-6 expression in 94 paraffin samples of gastric cancer tissues was examined by immunohistochemistry and its positivity was compared among different clinicopathologic features.

RESULTSFluorescent quantification PCR revealed gastric cancer tissues had significantly higher NEK-6 expression than para-cancer tissues(0.002 80±0.001 36 vs. 0.001 91±0.001 48, P<0.05), NEK-6 expression was up-regulated in 31 gastric cancer tissues (86.1%), and human gastric cancer cell lines had significantly higher NEK-6 expression than GES-1 cells, among whom BGC-823 and AGS cell lines were the highest. Invasion and migration tests showed that as compared to negative siRNA control group, ability of invasion and migration in BGC-823 and AGS cells after siRNA transfection was obviously decreased. In 94 paraffin samples, positive expression rate of NEK-6 was 60.6%(57/94), and NEK-6 expression was significantly associated with gastric cancer distant metastasis, lymph nodes metastasis and TNM staging(all P<0.05).

CONCLUSIONSNEK-6 expression is up-regulated in gastric cancer tissues, which is significantly associated with distant metastasis, lymph nodes metastasis and TNM staging. Down-regulation of NEK-6 expression can inhibit the ability of invasion and migration in gastric cancer cells.

Objective To explore the NEK-6 expression in gastric cancer tissue and its relationship with clinicopathological features. Methods Fluorescent quantification PCR and Western blotting were used to examine the NEK-6 expression in 36 samples of fresh gastric cancer tissues and para-cancer gastric mucosal tissues, human gastric cancer cell lines ﹙BGC-823, MKN-28, SGC-7901, MGC-803, HGC-27, AGS), and human normal gastric epithelial cell line ﹙GES-1). Gastric cancer cell lines with the highest expression level were selected to perform the invasion and migration tests, and the effect of down-regulated NEK-6 expression by siRNA transfection on above invasion and migration tests were observed. Meanwhile NEK-6 expression in 94 paraffin samples of gastric cancer tissues was examined by immunohistochemistry and its positivity was compared among different clinicopathologic features. Results Fluorescent quantification PCR revealed gastric cancer tissues had significantly higher NEK-6 expression than para-cancer tissues ﹙0.002 80 ±0.001 36 vs. 0.001 91 ±0.001 48, P<0.05), NEK-6 expression was up-regulated in 31 gastric cancer tissues ﹙86.1%), and human gastric cancer cell lines had significantly higher NEK-6 expression than GES-1 cells, among whom BGC-823 and AGS cell lines were the highest. Invasion and migration tests showed that as compared to negative siRNA control group, ability of invasion and migration in BGC-823 and AGS cells after siRNA transfection was obviously decreased. In 94 paraffin samples, positive expression rate of NEK-6 was 60.6%﹙57/94),and NEK-6 expression was significantly associated with gastric cancer distant metastasis, lymph nodes metastasis and TNM staging ﹙all P<0.05). Conclusions NEK-6 expression is up-regulated in gastric cancer tissues, which is significantly associated with distant metastasis, lymph nodes metastasis and TNM staging. Down-regulation of NEK-6 expression can inhibit the ability of invasion and migration in gastric cancer cells.

Objective To explore the NEK-6 expression in gastric cancer tissue and its relationship with clinicopathological features. Methods Fluorescent quantification PCR and Western blotting were used to examine the NEK-6 expression in 36 samples of fresh gastric cancer tissues and para-cancer gastric mucosal tissues, human gastric cancer cell lines ﹙BGC-823, MKN-28, SGC-7901, MGC-803, HGC-27, AGS), and human normal gastric epithelial cell line ﹙GES-1). Gastric cancer cell lines with the highest expression level were selected to perform the invasion and migration tests, and the effect of down-regulated NEK-6 expression by siRNA transfection on above invasion and migration tests were observed. Meanwhile NEK-6 expression in 94 paraffin samples of gastric cancer tissues was examined by immunohistochemistry and its positivity was compared among different clinicopathologic features. Results Fluorescent quantification PCR revealed gastric cancer tissues had significantly higher NEK-6 expression than para-cancer tissues ﹙0.002 80 ±0.001 36 vs. 0.001 91 ±0.001 48, P<0.05), NEK-6 expression was up-regulated in 31 gastric cancer tissues ﹙86.1%), and human gastric cancer cell lines had significantly higher NEK-6 expression than GES-1 cells, among whom BGC-823 and AGS cell lines were the highest. Invasion and migration tests showed that as compared to negative siRNA control group, ability of invasion and migration in BGC-823 and AGS cells after siRNA transfection was obviously decreased. In 94 paraffin samples, positive expression rate of NEK-6 was 60.6%﹙57/94),and NEK-6 expression was significantly associated with gastric cancer distant metastasis, lymph nodes metastasis and TNM staging ﹙all P<0.05). Conclusions NEK-6 expression is up-regulated in gastric cancer tissues, which is significantly associated with distant metastasis, lymph nodes metastasis and TNM staging. Down-regulation of NEK-6 expression can inhibit the ability of invasion and migration in gastric cancer cells.

Objective To investigate the profile of antimicrobial resistance in clinical isolates from the patients in Peking Union Medical College Hospital during 2012.Methods A total of 6 662 nonduplicate clinical isolates were collected.Disc diffusion test or Kirby-Bauer method and automated systems were employed to study the antimicrobial resistance.The data were analyzed by WHONET 5.6 software according to CLSI 2012 breakpoints.Results Of the 6 662 bacterial strains included in this analysis, gram negative organisms and gram positive cocci accounted for 66.7% (4 446/6 662)and 33.3% (2 216/6 662),respectively. The top 10 most frequently isolated microorganisms were E.coli (17%),P .aeruginosa (11.4%),A.baumannii (11.4%), S.aureus (11.2%),K.pneumoniae (9.2%),E.faecalis (8.4%),E.faecium (4.1%),coagulase negative Staphylococcus (3.3%),E.cloacae (3.1%)and S.maltophilia (3.1%).About 39.9% of the S.aureus strains and 73.4% of the coagulase negative Staphylococcus were methicillin-resistant.No staphylococcal strains were found resistant to vancomycin,teicoplanin or linezolid.A few of vancomycin-or teicoplanin-resistant strains were identified in both E.faecium and E.faecalis.No lin-ezolid resistant strains were found.ESBLs-producing strains accounted for 53.0%,25.7% and 27.0% in E.coli,Klebsiella spp.(K.pneumoniae and K.oxytoca)and P .mirabilis, respectively.The Enterobacteriaceae strains were still highly susceptible to carbapenems. Overall, less than 2.6% of these strains were resistant to carbapenems.A few pan-re-sistant strains of K.pneumoniae (0.7%,4/615)were iden-tified.About 20.3% and 13.6% of the P .aeruginosa isolates were resistant to imipenem and meropenem,respectively.P . aeruginosa isolates showed the lowest resistance rate (7.2%)to amikacin.And 72.8% and 75.2% of A.baumannii strains were resistant to imipenem and meropenem.A.baumannii isolates showed relatively low resistance rate to cefoperazone-sulbac-tam (51.2%)and minocycline (30.2%).The prevalence of pan-resistant strains was 43.5% in A.baumannii and 1.4% in P . aeruginosa.Conclusions Bacterial resistance is still increasing,especially pan-resistant A.baumannii strains.It is mandatory to take effective measures to control hospital infections and improve rational antibiotic use.

Objective:To analyse the biological function of anti-IL-6Rβ(gp130) monoclonal antibody and its regulatory effect on IL-6 signaling.Methods:Biological characteristics of anti-IL-6Rβ(gp130) mAb were assessed by Western blot analysis, capture ELISA and peptide ELISA .The phosphorylation of STAT 3 was tested by Western blot analysis in IL-6-stimulated U266/RA-FLS/RA-PBMC with or without anti-IL-6Rβ(gp130) mAb treatment.Results:3 strains of mouse anti-human gp130 mAb were with high affini-ty and different binding epitopes , the kaff of 10A1 was 2.62E-10.In U266, RA-PBMC and RA-SFMC, IL-6 signaling highly activated STAT3 which could be inhibited by anti-gp130 mAb.Conclusion: Anti-IL-6Rβ( gp130 ) mAb might have different binding epitopes and could affect IL-6 stimulated phosphorylation of STAT3, which provides a preliminary experiment for analyse the correlation of IL-6 signaling and RA .

Objective To study regulatory mechanism of osteopontin (OPN) in rheumatoid arthritis (RA). Methods The expression of OPN in peripheral blood mononuclear cells (PBMC), synovial fluid cells (SFMC) and synovium tissue (ST) and T cell subsets from RA patients were detected by real time PCR. The concentration and relative rate of inflammatory factors in the synovial fluid from RA patients were analyzed by ELISA. Results The mRNA expression of OPN in synovial fluid and tissue was higher than that of PBMC in the same RA patient. The OPN expression was found mainly on CD4+T. The OPN concentration was higher in the synovial fluid than that of in the same patient′s serum. Meanwhile, the concentration of IL-10, IFN-? and TNF-? was higher than that of in the serum from same patient. Also, the concentration of IL-18 and IL-12 were higher than that of normal individual serum. Conclusion OPN may control secretion of inflammatory factors of synovium tissue and synovial fluid and induce the inflammatory response.

Objective To investigate the antimicrobial resistance proifle in the clinical bacterial strains isolated from Peking Union Medical College Hospital during 2014.Methods A total of 8 295 nonduplicate clinical isolates were collected. Disc diffusion test (Kirby-Bauer method) and automated systems were employed to study the antimicrobial susceptibility. The data were analyzed by using WHONET 5.6 software according to CLSI 2014 breakpoints.Results Of the 8 295 isolates, 67.4% were gram-negative, and 32.6% were gram-positive. The top 10 most frequently isolated bacteria were:E. coli(18.1%),P. aeruginosa (10.8%),K. pneumoniae (10.2%),S. aureus (9.8%),
A. baumannii(9.2%),E. faecalis (6.3%),E. faecium (4.1%), coagulase-negativeStaphylococcus (4.1%),E. cloacae (3.1%) andS. maltophilia (2.9%). Methicillin resistant strains inS. aureus (MRSA) and coagulase negativeStaphylococcus (MRCNS) accounted for average of 28.4% and 66.5%, respectively. The resistance rates of MR strains to β-lactams and other antimicrobial agents were much higher than those MS strains. Overall, 81.3% of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole, while 81.1% of MRCNS strains were susceptible to rifampin. No staphylococcal strains were resistant to vancomycin, teicoplanin or linezolid. The resistance rate ofE. faecalis strains to most of the drugs tested (except chloramphenicol) was much lower than those ofE. faecium. Several strains of bothE. faecium andE. faecalis were found resistant to vancomycin and teicoplanin, which were Van-A and Van-B types based on their phenotype. No linezolid resistant enterococcal strains were found. Data showed that 90.8% ofβ-hemolyticStreptococcus strains were susceptible to penicillin. ESBLs-producing strains accounted for 54.2%, 31.0% and 28.9% inE. coli,Klebsiella spp (K. pneumoniae andK. oxytoca) andP. mirabilis, respectively.Enterobacteriaceae isolates were still highly susceptible to carbapenems. Overall, no more than 3.3% of these strains were resistant to carbapenems. A few extensively drug-resistant strains ofK. pneumoniae (1.3%, 11/842) were identiifed. The resistance rates ofP. aeruginosa to imipenem and meropenem were 17.5% and 11.8%, respectively.P. aeruginosa isolates showed the lowest resistance rate (5.9%) to amikacin. And 69.0% and 67.4% ofA. baumanniiisolates were resistant to imipenem and meropenem.A. baumannii isolates showed the lowest resistance rates to cefoperazone-sulbactam and minocycline (47.8% and 28.7%), respectively. The prevalence of extensively drug-resistant strains was 32.3% inA. baumannii and 1.8% inP. aeruginosa. The prevalence of β-lactamase inH. inlfuenzae was 33.7%. More than 93.0% ofS. pneumoniae strains were resistant to erythromycin and clindamycin.Conelusions Bacterial resistance is still increasing in this hospital, especially carbapenem resistantEnterobacteriaceae. It is necessary to take effective hospital infection control measures and use antibiotics rationally.

OBJECTIVE: To explore the genetic etiology for a Chinese pedigree affected with Alazami syndrome. METHODS: Genomic DNA was extracted for 2 patients and 2 unaffected members from the pedigree. Whole exome sequencing was carried out to detect potential variant in the proband, and the result was verified by Sanger sequencing. RESULTS: The proband and her sister were both found to harbor compound heterozygous variants of LARP7 gene, namely c.94A>T (p.Lys32*) and c.1141A>G (p.Lys381Glu), which were inherited from their father and mother, respectively. Both variants were predicted to be pathogenic based on bioinformatic analysis. CONCLUSION The two variants of the LARP7 gene, both were unreported previously, probably underlay the Alazami syndrome in this pedigree. Above finding has expanded the mutational spectrum of the LARP7 gene.
China ; Dwarfism ; Female ; Humans ; Intellectual Disability/genetics* ; Mutation ; Pedigree ; Ribonucleoproteins/genetics* ; Exome Sequencing

OBJECTIVE To investigate in vitro activities of 12 antimicrobial agents including cefmetazole against extended-spectrum beta-lactamases-producing Escherichia coli(528 strains),Klebsiella pneumoniae(311 strains) and Proteus mirabilis(15 strains).METHODS They all collected from 15 teaching hospitals in China during 2005 and 2006 and included in the study.The levels of minimal inhibitory concentration(MIC) of 12 antimicrobial agents were determined by agar dilution method.WHONET 5.4 Software was used to analyze the data.RESULTS Against ESBLs-producing E.coli and ESBLs-producing K.pneumoniae,carbapenems were the most active antimicrobial agents(all 100.0% susceptible),followed by cephamycins(80.1-97.3%).Piperacillin/tazobactam(78.5-95.1%)showed a higher activity than cefoperazone/sulbactam(44.1-56.2%).The susceptible rate to ceftazidime against ESBLs-producing E.coli was remarkably higher than the other three cephalosporins,however the differences did not happen to ESBL-producing K.pneumoniae obviously.The susceptible rate to cefuroxime was below 1.6%.ESBLs-producing K.pneumoniae showed high sensitivity to carbapenems,cephamycins and ?-lactam/lactamase inhibitor combinations(all 100% susceptible),however the susceptible rates to cephalosporins were relatively lower.CONCLUSIONS Carbapenems and cephamycins remain the relatively high activity against ESBLs-producing Enterobacteriaceae.

Objective To evaluate the influences of susceptibility interpretation of Escherichia coli,Klebsiella pneumonia and Proteus mirabilis in China mainland according to the old and new ceftazidime,cefotaxime and ceftriaxone breakpoints in CLSI M100-S20 and CLSI M100-S19. Methods First, We analyzed the antibacterial susceptibility results of the three bacteria by agar dilution method in the SEANIR surveillance item, which were collected from 15 national hospitals between the year of 2005 and 2007 and excluded the AmpC enzyme positive isolates according to the PGR-DNA sequencing method and/or the antibacterial susceptibility phenotype. ESBL phenotype was confirmed by the CLSI phenotypic confirmatory test. Antibacterial susceptibility of the total 2733 Escherichia coli, Klebsiella pneumonia, Proteus mirabilis isolates was retrospectively analyzed by WHONET 5. 4 software according to the breakpoints of the CLSI M100-S19 (S19) and CLSI M100-S20 (S20). Second, 207 isolates of Peking Union Medical College Hospital with the results of both agar dilution method and disk diffusion method were performed by recurrent analysis. Then we observed the inter-method agreement through the scatter diagram according to the breakpoints of S19 and S20. Results First, as to the ESBL positive Escherichia coli, Klebsiella pneumonia and Proteus mirabili.s, the resistant rate of cefotaxime increased from 65.2% , 55.5%, 14. 6% under S19 (64 μg/ml) to 99. 7%, 96. 2% , 93. 8% under S20 (4 μg/ml). The susceptibility rates decreased from 6. 0%, 11.5%, 33.3% under S19 (8 μg/ml) to 0%, 0. 2%, 0% under S20 ( 1 μg/ml). Ceftriaxone had the same trend as cefotaxime. Though ceftazidime was more active than cefotaxime and ceftriaxone, as to the ESBL positive Escherichia coli and Klebsiella pneumonia, the resistant rates slightly increased from 30. 3%,43. 2% under S19 (32 μg/ml) to42.0%, 56. 0% under S20 (16 μg/ml). The susceptibility rates slightly decreased from 58. 1%, 44. 1% under S19 (8 μg/ml) to 44. 7%, 28.0% under S20 (4 μg/ml). Second,as to the ESBL negative Escherichia coli, Klebsiella pneumonia and Proteus mirabilis, all the susceptibility rates of ceftazidime, cefotaxime and ceftriaxone were between 99. 2%-100. 0%, the resistant rate were between 0%-0. 4%. Third, the S20 MIC breakpoints had a good correspondence with the ESBL phenotype.Fourth, according to the recurrent analysis of MIC testing and disk dilution method, r value was 0. 67,0. 79, 0. 77 for ceftazidime, cefotaxime and ceftriaxone, respectively, and all P value were under 0. 01. The intermethod rates of S19 and S20 were both acceptable. Conclusions If the cefotaxime and ceftriaxone S20 new breakpoints were used, the concordance of antibacterial susceptibility results and ESBL phenotype would increase greatly. The clinician could select proper antibiotics according to the antibacterial susceptibility results and clinical symptoms. It is no longer necessary to edit results for cephalosporins, aztreonam, or penicillins from susceptible to resistant. However, until laboratories implement the new interpretive criteria,ESBL testing should be performed as described in Supplemental Table 2A-S1. The relationship between the new breakpoints of ceftazidime and clinical outcomes need to be further evaluated.