ABSTRACT
BACKGROUND@#Long non-coding RNA (lncRNA) actin filament-associated protein 1 antisense RNA 1 (AFAP1-AS1) functions as a competing endogenous RNA to regulate target genes expression by sponging microRNAs (miRs) to play cancer-promoting roles in cancer stem cells. However, the regulatory mechanism of AFAP1-AS1 in cervical cancer (CC) stem cells is unknown. The present study aimed to provide a new therapeutic target for the clinical treatment of CC.@*METHODS@#Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 (CD44v6)(+) CC cells were isolated by flow cytometry (FCM). Small interfering RNAs of AFAP1-AS1 (siAFAP1-AS1) were transfected into the (CD44v6)(+) cells. The levels of AFAP1-AS1 were measured by quantitative real-time PCR (qRT-PCR). Sphere formation assay, cell cycle analysis, and Western blotting were used to detect the effect of siAFAP1-AS1. RNA pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAP1-AS1 or vascular endothelial growth factor (VEGF)-C.@*RESULTS@#CD44v6(+) CC cells had remarkable stemness and a high level of AFAP1-AS1. However, AFAP1-AS1 knockdown with siAFAP1-AS1 suppressed the cell cycle transition of G(1)/S phase and inhibited self-renewal of CD44v6(+) CC cells, the levels of the stemness markers octamer-binding transcription factor 4 (OCT4), osteopontin (OPN), and cluster of differentiation 133 (CD133), and the epithelial-mesenchymal transition (EMT)-related proteins Twist1, matrix metalloprotease (MMP)-9, and VEGF-C. In the mechanism study, miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+) CC cells.@*CONCLUSIONS@#LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.
Subject(s)
Female , Humans , Cell Line, Tumor , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Uterine Cervical Neoplasms/genetics , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor CABSTRACT
The applicator therapy is a unique method to treat infant diarrhea in traditional Chinese medicines and widely applied in clinical practice. Currently, many researchers have proved the rationality of the therapy based on the traditional Chinese medicine mechanism and on the data from clinical practice, but its action mechanism is uncertain at present. In this study, with the assistance of pediatric practitioners, the automated ribosomal intergenic-spacer analysis (ARISA) was adopted to study the effect of the adjuvant therapy with Dingguier umbilical paste on intestinal flora of diarrhea infants, in which Dingguier umbilical paste served as the adjuvant therapy in oral traditional Chinese medicines and fecal samples of infants with different diarrhea symptoms were collected and used as the study materials. The results showed that the adjuvant therapy had a significant effect on the shift of intestinal flora, which was associated with the decrease in the similarity difference to the normal control group and the increase in the number of operational taxonomic units (OTUs) shared with the normal control group. Additionally, adjuvant therapy with Dingguier umbilical paste also showed long action duration and increased OTUs number. These results indicated that Dingguier umbilical paste has the effect in restoring the micro-ecosystem of unbalanced intestinal bacteria. Intestinal flora may be one of major targets for the applicator therapy for the infant diarrhea, but not for the single oral traditional Chinese medicine for infant diarrhea.
Subject(s)
Female , Humans , Infant , Male , Adjuvants, Pharmaceutic , Therapeutic Uses , Diarrhea , Drug Therapy , Microbiology , Feces , Microbiology , Intestines , Microbiology , Medicine, Chinese Traditional , Methods , Ointments , Treatment Outcome , UmbilicusABSTRACT
<p><b>OBJECTIVE</b>To investigate changes of two sodium channels, PN(3) and NaN, during orofacial pain by occlusal trauma in rat.</p><p><b>METHODS</b>Expressions of PN(3) mRNA and NaN mRNA in trigeminal ganglion were tested during various periods of persistent occlusal trauma with reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In groups, including control, occlusal trauma groups, PN(3) mRNA and NaN mRNA were all expressed in trigeminal ganglion neurons. In the control group, there were similar density values bilaterally. In the occlusal trauma group, the density values in gel electrophoresis of PN(3) mRNA and NaN mRNA on the intervention side were slightly greater than those on the control side.</p><p><b>CONCLUSIONS</b>The stimulation of occlusal trauma upregulates expressions of PN(3) mRNA and NaN mRNA, which suggests the signal occurring and conduction of chronic pain by occlusal trauma have the same molecular mechanism of sodium channel as inflammatory pain.</p>
Subject(s)
Animals , Male , Rats , Dental Occlusion, Traumatic , Facial Pain , RNA, Messenger , Rats, Sprague-Dawley , Sodium Channels , Genetics , Trigeminal Ganglion , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effect of traumatic occlusion on CGRP-immunoreactive (CGRP-IR) nerve fibres in rat molar pulp and observe the recovery of CGRP-IR nerve fibres after removal of traumatic occlusion.</p><p><b>METHODS</b>To observe immunohistochemically the change of CGRP-IR nerve fibres in molar pulp during traumatic occlusion and after removal.</p><p><b>RESULTS</b>The increase of number, density and morphology of CGRP-IR nerve fibres in traumatic occlusion group was more than in control group, however, the changes of CGRP-IR nerve fibres in removal of traumatic occlusion group were less than in control group.</p><p><b>CONCLUSIONS</b>The changes of CGRP-IR nerve fibres in number, morphology, and density are induced by traumatic occlusion in rat molar pulp, however, the nerve fibres recover to normal by removal of traumatic occlusion.</p>