First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2

BACKGROUND Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice.These cells consist of both epithelial and mesenchymal cells.Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma.However,cholangiocarcinoma sarcoma cell lines are not available in cell banks.AIM To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line,namely CBC2T-2.METHODS We conducted a short tandem repeat(STR)test to confirm the identity of the CBC2T-2 cell line.Furthermore,we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies.The tumorigenic potential of CBC2T-2 cells was tested in vivo using nonobese diabetic/severe combined immunodeficient(NOD/SCID)mice.The cells were injected subcutaneously and tumor formation was observed.In addition,immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts.The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma.Lastly,whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line.RESULTS The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue.The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology.The cells exhibited a high proliferation ratio with a doubling time of 47.11 h.This cell line has migratory,invasive,and clonogenic abilities.The chromosomes in the CBC2T-2 cells were polyploidy,with numbers ranging from 69 to 79.The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice.CBC2T-2 cells and xenografts were positive for both the epithelial marker,CK19,and the mesenchymal marker,vimentin.These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics.The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma,and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option.CONCLUSION We established the first human cholangiocarcinoma sarcoma cell line,CBC2T-2,with stable biogenetic traits.This cell line,as a research model,has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.

Functional types of long trichoid sensilla responding to sex pheromone components in Plutella xylostella

Sex pheromones,which consist of multiple components in specific ratios pro-mote intraspecific sexual communications of insects.Plutella xylostella(L.)is a world-wide pest of cruciferous vegetables,the mating behavior of which is highly dependent on its olfactory system.Long trichoid sensilla on male antennae are the main olfactory sen-silla that can sense sex pheromones.However,the underlying mechanisms remain unclear.In this study,3 sex pheromone components from sex pheromone gland secretions of P xy-lostella female adults were identified as Z11-16:Ald,Z11-16:Ac,and Z11-16:0H in a ratio of 9.4:100:17 using gas chromatography-mass spectrometry and gas chromatography with electroantennographic detection.Electrophysiological responses of 581 and 385 long trichoid sensilla of male adults and female adults,respectively,to the 3 components were measured by single sensillum recording.Hierarchical clustering analysis showed that the long trichoid sensilla were of 6 different types.In the male antennae,52.32%,5.51%,and 1.89%of the sensilla responded to Z11-16:Ald,Z11-16:Ac,and Z11-16:0H,which are named as A type,B type,and C type sensilla,respectively;2.93%named as D type sen-silla responded to both Z11-16:Ald and Z11-16:Ac,and 0.34%named as E type sensilla were sensitive to both Z11-16:Ald and Z11-16:OH.In the female antennae,only 7.53%of long trichoid sensilla responded to the sex pheromone components,A type sensilla were 3.64%,B type and C type sensilla were both 0.52%,D type sensilla were 1.30%,and 1.56%of the sensilla responded to all 3 components,which were named as F type sen-silla.The responding long trichoid sensilla were located from the base to the terminal of the male antennae and from the base to the middle of the female antennae.The pheromone mixture(Z11-16:Ald:Z11-16:Ac:Z11-16:0H=9.4:100:17)had a weakly repellent effect on female adults of P xylostella.Our results lay the foundation for further studies on sex pheromone communications in P.xylostella.

Recent Progress in Carbon-based Materials of Non-Noble Metal Catalysts for ORR in Acidic Environment

Proton exchange membrane fuel cell(PEMFC)has important implications for the success of clean transportation in the future.One of the key factors affecting the cost and performance of PEMFC is the cathode electrocatalyst for the oxygen reduction reaction(ORR)to overcome sluggish kinetics and instability in an acidic environment.As an essential component of the electrocatalyst,the support material largely determines the activity,mass transfer,charge transfer,and durability of the electrocatalyst.Thereby,the support material plays a critical role in the overall performance of the electrocatalyst.Carbonbased materials are widely used as electrocatalyst supports because of their high porosity,conductivity,chemical stability,and tunable morphology.Recently,some new carbon-based materials with excellent structure have been introduced,such as carbon nanotubes,carbon nanowires,graphene,metal-organic framework(MOF)-derived carbon,and biomass-derived carbon materials.Combined with a variety of strategies,such as controllable construction of porous structures and surface defects,proper doping heteroatoms,the ingenious design of model electrocatalysts,and predictive theoretical calculation,a new reliable path was provided for further improving the performance of electrocatalysts and exploring the catalytic mechanism.Based on the topic of carbon-based materials for ORR in acidic medium,this review summarizes the up-to-date progress and breakthroughs,highlights the factors affecting the catalytic activity and stability of ORR electrocatalysts in acids,and discusses their future application and development.