OBJECTIVEThis investigation aimed to examine how buccal mucosa microbiome succeeds in a healthy population with different ages and dentition stages.

METHODSTwenty-five subjects were recruited and subdivided into five groups: primary dentition group, mixed dentition group, adolescent group, adult group, and elderly group. Individual mucosal microbiota was obtained by gently scraping both sides of the buccal mucosa with a cotton swab. Microbial diversity was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE).

RESULTS1) The composition of buccal mucosa microbiota has great intra-individual divergence. 2) The average band numbers of the primary dentition group, mixed dentition group, adolescent group, adult group, and elderly group were 21.2 +/- 4.0, 17.8 +/- 3.9, 15.8 +/- 4.3, 16.8 +/- 3.7, and 22.2 +/- 6.5, respectively. No between-group differences was observed (P > 0.05), indicating that predominant strains in the oral cavity may be stable throughout an individual's lifetime. 3) The Shannon indices of primary dentition group, mixed dentition group, adolescent group, adult group, and elderly group were 1.73 +/- 10.2, 1.43 +/- 0.1, 1.05 +/- 0.2, 1.45 +/- 0.2, and 1.63 +/- 0.3, respectively. A significant between-group difference was observed (P = 0.003), indicating that the microbial diversity of the buccal mucosa decreases from childhood through adolescence, but increases from adult through senescence. 4) The clustering analysis showed that most of the samples in the same group clustered together, indicating higher intra-group community structure similarity.

CONCLUSIONComposition of the buccal mucosa microbiota was different among age groups. Adolescence may be an essential turning point of microbial ecology succession throughout life.

Adolescent ; Adult ; Aged ; DNA, Bacterial ; Humans ; Microbiota ; Mouth Mucosa ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S

AIM:To investigate the effect of plate-let(PLT)on angiogenesis in hepatocellular carcino-ma and the intervention effect of Cinobufagin(CBG).METHODS:Firstly,we screened the suitable co-incubation ratio of PLT and hepatocellular carci-noma cells,prepared conditioned medium,and de-termined the half inhibitory concentration of Cinob-ufagin;then,we set up a control group(human um-bilical vein endothelial cells(EC)+conventional me-dium),a crosstalk group(EC+CM_HP(strip culture prepared by crosstalk of HUH7 and PLT)),and an in-tervention group(EC+CM_HP+CBG).The migration,tube-formation and sprouting capacity of EC and the level of vascular endothelial growth factor(VEGF)in co-cultured supernatant were evaluated by scratch assay,tube-formation assay,budding as-say and ELISA assay.Western blot was used to de-tect the expression of VEGFR2 and p-VEGFR2,and reverse verification was performed with inhibitors.A subcutaneous transplantation tumour model of hepatocellular carcinoma in nude mice was estab-lished,with Model group,Model+CBG group and Model+Apa group.The collagen expression of the transplantation tumour was observed by Masson staining,and the expression levels of vascular endo-thelial markers CD31 and CD34 were detected by immunofluorescence.RESULTS:When PLT:HUH7=200,the activity of HUH7 was the strongest,and the crosstalk between HUH7 and PLT significantly promoted the proliferation of EC(P＜0.01).Com-pared with Control group,the migration,tube-for-mation and budding ability of Crosstalk group were enhanced,and those of Intervention group were lower than those of Crosstalk group(P＜0.01).The expression level of VEGF in the supernatant of Crosstalk group was higher than that of Control group,while that of Intervention group was lower than that of Crosstalk group(P＜0.01).The expres-sion level of p-VEGFR2 protein in Crosstalk group was significantly higher than that of Control group,but the expression level of Intervention group was lower than that of Crosstalk group(P＜0.01).Large collagen fibre deposition was seen in the Model group,and CBG intervention significantly reduced collagen fibre deposition in the transplanted tu-mour tissues.CD31 and CD34 expression was pres-ent in the hepatocellular carcinoma transplanted tumour tissues in the Model group,and CBG inter-vention significantly reduced the expression of CD31 and CD34 in the liver cancer transplanted tu-mour tissues(P＜0.01).CONCLUSION:PLT enhances angiogenesis in hepatocellular carcinoma,and CBG may inhibit its tube-forming ability via the VEGF/VEGFR2 pathway.

Objective This investigation aimed to examine how buccal mucosa microbiome succeeds in a healthy population with different ages and dentition stages. Methods Twenty-five subjects were recruited and subdivided into five groups: pri-mary dentition group, mixed dentition group, adolescent group, adult group, and elderly group. Individual mucosal microbiota was obtained by gently scraping both sides of the buccal mucosa with a cotton swab. Microbial diversity was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Results 1) The composition of buccal mucosa microbiota has great intra-individual divergence. 2) The average band numbers of the primary dentition group, mixed dentition group, adolescent group, adult group, and elderly group were 21.2±4.0, 17.8±3.9, 15.8±4.3, 16.8±3.7, and 22.2±6.5, respectively. No between-group differences was observed (P>0.05), indicating that predominant strains in the oral cavity may be stable throughout an individual’s lifetime. 3) The Shannon indices of primary dentition group, mixed dentition group, adolescent group, adult group, and elderly group were 1.73±0.2, 1.43±0.1, 1.05±0.2, 1.45±0.2, and 1.63±0.3, respectively. A significant between-group difference was observed (P=0.003), indicating that the microbial diversity of the buccal mucosa decreases from childhood through adolescence, but increases from adult through senescence. 4) The clustering analysis showed that most of the samples in the same group clustered together, indicating higher intra-group community structure similarity. Conclusion Composition of the buccal mucosa microbiota was different among age groups. Adolescence may be an essential turning point of microbial ecology succession throughout life.