Objective To uncover and identify the differences in salivary microbiota profiles and their potential roles between patients with pulmonary nodules(PN)and healthy controls,and to propose new candidate biomarkers for the early warning of PN.Methods 16S rRNA amplicon sequencing was performed with the saliva samples of 173 PN patients,or the PN group,and 40 health controls,or the HC group,to compare the characteristics,including diversity,community composition,differential species,and functional changes of salivary microbiota in the two groups.Random forest algorithm was used to identify salivary microbial markers of PN and their predictive value for PN was assessed by area under the curve(AUC).Finally,the biological functions and potential mechanisms of differentially-expressed genes in saliva samples were preliminarily investigated on the basis of predictive functional profiling of Phylogenetic Investigation of Communities by Reconstruction of Unobserved States(PICRUSt2).Results The α diversity and βdiversity of salivary microbiota in the PN group were higher than those in the HC group(P＜0.05).Furthermore,there were significant differences in the community composition and the abundance of oral microorganisms between the PN and the HC groups(P＜0.05).Random forest algorithm was applied to identify differential microbial species.Porphyromonas,Haemophilus,and Fusobacterium constituted the optimal marker sets(AUC=0.79,95％confidence interval:0.71-0.86),which can be used to effectively identify patients with PN.Bioinformatics analysis of the differentially-expressed genes revealed that patients with PN showed significant enrichment in protein/molecular functions involved in immune deficiency and redox homeostasis.Conclusion Changes in salivary microbiota are closely associated with PN and may induce the development of PN or malignant transformation of PN,which indicates the potential of salivary microbiota to be used as a new non-invasive humoral marker for the early diagnosis of PN.

MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA bio-logical function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and spec-ificity, and applicability to disease diagnosis and therapy.

Objective To investigate the promotion effect of Danhong injection (DH) on brain-derived neurotrophic factor (BDNF) expression in Schwann cells (SCs) of SD rats.Methods In experiment of SCs apoptosis induced by advanced glycation end products (AGEs),SCs were divided into control group,AGEs treatment group and DH+AGEs treatment group; 48 h after each treatment,the SCs count was compared.In experiment of DH affecting mRNA and protein BDNF expressions in SCs,real time-PCR and Western blotting were used.In the experiment of DH combined with different inhibitors (Calphostin C,LY294002,H89,U0126,FR180204 and SB203580) affecting mRNA BDNF expression in SCs,real time-PCR was used.Results The number of SCs in AGEs treatment group was significantly decreased than that in the control group,but that in DH+AGEs treatment group was statistically increased than that in AGEs group (P＜0.05).The mRNA and protein expressions of BDNF in the DH treatment group were significantly increased than those in the control group (P＜0.05).As compared with DH group,DH+Calphostin C treatment group had significantly decreased BDNF mRNA expression (P＜0.05); BDNF mRNA expression in the DH+U0126,DH+FR180204 and DH+SB203580 treatment groups was significantly decreased as compared with that in the DH treatment group (P＜0.05).Conclusions DH could effectively inhibit SCs apoptosis induced by AGEs and significantly promote BDNF expression;protein kinase C (Calphostin C) and mehtyl ethyl ketone (U0126)/extracellular regulated protein kinases (FR180204EK)/p38 (SB203580) may be the important signaling transconduction pathways for BDNF expression.