Résumé
This study was aimed at identifying species of ticks and investigating infections with tick-borne pathogens a-mong people in Yadong,Tibet.The total of 23 ticks(Ixodes and Haemaphysalis)collected from livestock in Yadong,Tibet in July 2021,the gene sequences of cytochrome oxidase subunitⅠ(COX Ⅰ),mitochondrial 16S ribosomal DNA(16S rDNA)and mitochondrial 12S ribosomal DNA(12S rDNA)were identi-fied on the basis of molecular biology classification.The gene sequences of spotted fever group rickettsia(SFGR),Borrelia burgdorferi,Francisella tularensis and Coxiella burnetii in ticks were amplified by PCR and sequenced,and phylogenetic ana-lyses were performed.In addition,184 serum samples collected from Yadong in 2021 were screened for antibodies to Lyme dis-ease,Q fever,spotted fever,and tularemia by ELISA,and infections with tick-borne pathogens in Yadong were comprehen-sively analyzed.The ticks(Ixodes)collected in Yadong,Tibet,were grouped into two categories.The COX Ⅰ,16S rDNA,and 12S rDNA gene sequences of the ticks had 99.5％,97.57％,and 99.12％ similarity,respectively,to those of Ixodes nuttallia-nus in GenBank.The similarities between the COX Ⅰ and 16S rDNA gene sequences of another Ixodes tick species and the COXⅠ and 16S rDNA gene sequences of Ixodes ovatus in GenBank were 88.29％ and 95.75％,respectively.The similarity between the COX Ⅰ,16S rDNA,and 12S rDNA gene sequences of Haemaphysalis ticks and those of Haemaphysalis Tibet in GenBank were 96.04％,96.17％,and 97.47％,respectively.The sequence amplification results for Borrelia burgdorferi,Francisella tularensis,and Coxsiella burnetii in 23 ticks were negative,whereas gltA gene amplification of spotted fever group Rickettsia in 23 ticks was positive.The similarity of gltA gene sequences with respect to those of Candidatus Rickettsia tarasevichiae,Rickettsia raoultii,Rickettsia canadensis,and a Rickettsia endosymbiont of Eucalyptus brunneri in GenBank was 96.77％,96.76％,95.35％,and 95.35％,respectively.Among 184 serum samples from Yadong,the positivity rates of Lyme disease,Q fever,spotted fever,and tularemia were 6.52％,12.50％,8.70％,and 10.87％,respectively.These results indicated that the 23 parasitic ticks collected in Yadong,Tibet,in July 2021 were Ixodes nuttallianus,Ixodes ovatus and Haemaphysalis ti-betensis,and 23 ticks were infected with SFGR.Lyme disease,Q fever,spotted fever,and tularemia infection were prevalent in the population of Yadong.Therefore,efforts to control tick-borne diseases must be strengthened.
Résumé
Objective: To investigate the effect of poly (L-lactic acid caprolactone) (PLCL) /gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs). Methods: Rat bone marrow-derived EPCs were isolated and cultured, then identification was performed. After preparation of PLCL/gelatin blend electrospun scaffold, scanning electron microscopy and water contact angle test were carried out. EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation. The expression of vascular endothelial growth factor (Vegf ) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting. Results: The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs. PLCL/gelatin electrospun nanofibers were porous, and the hydrophilic properties were favorable for cell adhesion, and EPCs grew well on the scaffold. The expression of Vegf and Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000), and the expression of VEGF protein was also increased (P=0.000). Conclusion: PLCL/gelatin is an ideal scaffold for tissue engineering, and it can promote the angiogenesis differentiation of EPCs.
Résumé
Objective To study and prepare the monoclonal antibody library against human FXYD6 functional region,to screen the hybridoma cell lines secreting the monoclonal antibodies against intracellular or extracellular region of human FXYD6,and to identify the biological function of monoclonal antibody against extracellular domain.Methods FXYD6 functional region recombinant protein which did not contain the transmembrane region was prokaryotically expressed,purified,and FXYD6 recombinant protein was used to immunize BALB/c mice.Then splenocytes after immunization were fused with myeloma cells SP2/0.After several rounds of screening and cloning,the hybridomas which secreted the antibodies against the extracellular domain or the intracellular domain of human FXYD6 were established.The antibody specificity and subtype were identified with indirect ELISA,western blot and immunohistochemistry.The monoclonal antibodies against the extracellular domain which recognized the native conformation were screened with flow cytometry.The antibody against extracellular region was prepared with the ascites revulsion method and purified.The affinity constants were measured with indirect ELISA.The function of extracellular monoclonal antibody was detected by HepG2 cell line with high expression of FXYD6.Results The hybridoma cell library which secreted the monoclonal antibody against extracellular domain or the intracellular domain of human FXYD6 was successfully obtained,and extracellular region monoclonal antibodies with the functional blocking were prepared.Conclusion The prepared anti-human FXYD6 extracellular monoclonal antibodies could inhibit HepG2 cell proliferation.
Résumé
Objective · To investigate the effect of poly (L-lactic acid caprolactone) (PLCL)/gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs).Methods· Rat bone marrow-derived EPCs were isolated and cultured,then identification was performed.After preparation of PLCL/gelatin blend electrospun scaffold,scanning electron microscopy and water contact angle test were carried out.EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation.The expression of vascular endothelial growth factor (Vegf) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting.Results· The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs.PLCL/gelatin electrospun nanofibers were porous,and the hydrophilic properties were favorable for cell adhesion,and EPCs grew well on the scaffold.The expression of Vegfand Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000),and the expression of VEGF protein was also increased (P=0.000).Conclusion · PLCL/gelatin is an ideal scaffold for tissue engineering,and it can promote the angiogenesis differentiation of EPCs.