Aim TostudytheeffectsofCuO,ZnOand TiO2 nanoparticles on the viability and metastatic po-tential of EC-9706 and EC-109 esophageal squamous carcinomacelllineinvitro.Methods Characteristics of CuO,ZnO and TiO2 nanoparticles were detected u-sing transmission electron microscope (TEM)and dy-namic light scattering (DLS ).EC-9706 and EC-109 cells were treated with different concentrations of CuO, ZnO and TiO2 (5 ~80 mg · L-1 ).The cell prolifera-tion was analyzed by MTT assay.The cell cycle and apoptotic rates were determined by flow cytometry (FCM).The cell invasion was assayed in Transwell chambers.The expression of Bcl-2 and caspase-3 pro-tein in cells was detected by Western blot method.Re-sults CuO,ZnOandTiO2nanoparticleswerespheri-cal with primary particle size 12,20. 6,12 nm.The particles were agglomerated in water and cell culture medium with negative charge.CuO and ZnO nanoparti-cles induced decreases in EC-9706 and EC-109 cell vi-ability dose-dependently.After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G2/Mand S phase,and up-regulation of the cells in G0/G1 phase.Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment. Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously (P<0. 05 )compared with control group.TiO2 nanop-articles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and inva-sion.Conclusion ComparedwithTiO2,CuOand ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G0/G1 phase and apoptosis.These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.

In order to explore the genomic characteristics,its virulence,evolution,the correlation be-tween drug resistance phenotype and resistance genes of S.equi XJ 5012,to laid a theoretical basis for efficient prevention and control of strangles,whole genome sequencing was investigated.In this study,the antimicrobial suceptibility of S.equi XJ 5012 was tested by the paper disk method and its whole genome was sequenced,Then the 16S rRNA evolution,SeM genotyping,antibiotic resist-ance gene and virulence gene were compared and analyzed.Drug susceptibility results showed that S.equi XJ5012 was sensitive to 13 antibiotics,such as amoxicillin,ampicillin,cefuroxime,cefotio-fur,cefoxitin,streptomycin,erythromycin,oxytetracycline,levofloxacin,norfloxacin,ciprofloxacin,rifampicin,clindamycin,sulfadiazine sodium,and resistant to 6 antibiotics,such as penicillin,genta-micin,clarithromycin,doxycycline,tetracycline and sulfonamide isoxazole.The whole genome se-quencing results of showed that the genome size of S.equi XJ 5012 was 2.21 Mb,the GC content was 41.31％and this strain has 2 264 code genes.Antibiotic resistance genes database(ARDB)in-dicated that the S.equi XJ 5012 carried 135 drug resistance gene,among which the resistance genes mainly were macrocyclic lipids,fluoroquinolones,aminoglycosides,peptides,tetracyclines and β-lac-tam antibiotics,correlating well with the results of drug susceptibility test.Virulence Factors Data-base(VFDB)indicated that S.equi XJ 5012 carried 197 virulence genes include adhesion,antiph-agocytosis,extracellular enzyme encoding,secrtion,and iron uptake.The phylogenetic tree based on 16S rRNA gene confirmed that the strain XJ 5012 was closely related to S.equi 4047.The results of SeM genotyping revealed that S.equi XJ 5012 was SeM-217,which was closely related to Xin-jiang isolates SeM-209 and SeM-215 and foreign isolates SeM-195 and SeM-196.In this study,the whole genome sequencing analysis of S.equi XJ 5012 based on the third generation sequencing technology is the first report on the genome of S.equi in China,which provides a reference for a more comprehensive and in-depth understanding the characteristics of the genomic,molecular epi-demic and drug resistance of the strain.It is significant for the further prevention and control of strangles and understanding of pathogenic mechanism of S.equi.

Objective:To explore the effect of esophageal squamous cell carcinoma-related long non-coding RNA (lncRNA) ESCCAL-1 on the polarization of THP-1 cells-derived macrophages.Methods:The esophageal cancer cell line KYSE450 was divided into 5 groups: KYSE450 group (normal KYSE450 cells), shRNA-ESCCAL-1 group (infected with knockout ESCCAL-1 lentivirus), shRNA-NC group (infected with interference control lentivirus), OE-ESCCAL-1 group (infected with overexpressing ESCCAL-1 lentivirus) and OE-NC group (infected with overexpressed control lentivirus). The expression of ESCCAL-1 was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After co-culture of cells in each group with THP-1 cells-derived macrophages, flow cytometry was used to detect the expressions of THP-1 cells-derived macrophages M1 polarization markers HLA-DR, iNOS, CD86 and M2 polarization markers Arg-1, CD163, CD206, and inflammatory cytokines.Results:After THP-1 cells were stimulated with 100 ng/ml phorbol ester for 48 hours, the cells grew adherently, and the expression levels of CD11b and CD36 increased, indicating that THP-1 cells were successfully differentiated into macrophages. After THP-1 cells-derived macrophages were co-cultured with esophageal cancer KYSE450 cell line treated differently for 24 hours, there were no significant differences in the expressions of M1 polarization markers HLA-DR, iNOS and CD86 between shRNA-ESCCAL-1 group and shRNA-NC group or between OE-ESCCAL-1 group and OE-NC group (all P > 0.05). Compared with shRNA-NC group, the expressions of M2 polarization markers Arg-1, CD163 and CD206 in shRNA-ESCCAL-1 group decreased [8.54±0.29 vs. 11.83±0.69, 12.0±0.3 vs. 24.5±0.8, 2.05±0.23 vs. 14.54±1.10], and the differences were statistically significant ( t values were 7.636, 27.38 and 19.31, all P < 0.01); compared with the OE-NC group, the expressions of M2 polarization marker Arg-1, CD163 and CD206 in OE-ESCCAL-1 group increased [32.60±1.14 vs. 14.20±0.20, 43.7±1.5 vs. 25.1±1.2, 35.8±0.7 vs. 13.6±0.6], and the differences were statistically significant ( t values were -27.58, -17.24 and -43.98, all P < 0.01). Compared with shRNA-NC group, the expression level of interferon-γ in shRNA-ESCCAL-1 group decreased [(6.3±1.5) pg/ml vs. (20.0±2.6) pg/ml, t = 7.75, P = 0.001]; compared with OE-NC group, the expression level of interleukin-1RA in OE-ESCCAL-1 group increased [(3 167±306) pg/ml vs. (467±176) pg/ml, t = -13.27, P < 0.01]. Conclusions:Esophageal squamous cell carcinoma-related lncRNA ESCCAL-1 can promote the M2 polarization of macrophages.

OBJECTIVETo observe the effects of manganese sulfate on blood pressure, myocardial ultrastructure and heart organ index of rats.

METHODSForty male SPF SD rats were randomly divided into 4 groups: control group (0 mg/kg), 5 mg/kg dose group, 15 mg/kg dose group and 25 mg/kg dose group, 10 rats each group. Intraperitoneal injection was performed for six months, by five times each week, the rat blood pressure was measured by tail cuff method, and the heart organ index of the rats was computed. Three rats were selected from each group randomly, and the myocardial ultrastructure of the rats was observed by using transmission electron microscopy (TEM). The BMD and BMDL between manganese sulfate injected dose and the rats heart organ index were evaluated by BMD (Benchmark Dose).

RESULTSThere was no significant of blood pressure between the experimental group and the control group (P > 0.05).The heart organ indexes of the four groups were 0.24% ± 0.10%, 0.25% ± 0.02%, 0.26% ± 0.02%, and 0.24% ± 0.02%. Statistical significance of heart organ indexes was found between the 15 mg/kg dose group and the control group (P < 0.05). Observed by TEM, we found that-different degrees of mitochondrial crest fracture or disappear, mitochondria swelling, hydropic change and myocardial fibers degeneration happened in the rats of the three exposed groups, but not the control group. The BMD and BMDL were calculated as 9.33 mg/kg and 4.28 mg/kg in the study of manganese sulfate injected dose and the rats heart organ index.

CONCLUSIONChronic manganese poisoning can lead to myocardial mitochondria superfine lesions, myocardial fiber damage and heart organ index change in rats.

Animals ; Male ; Manganese Compounds ; Mitochondria ; drug effects ; ultrastructure ; Myocardium ; ultrastructure ; Myocytes, Cardiac ; drug effects ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Sulfates ; toxicity ; Toxicity Tests