Four-Octyl itaconate ameliorates periodontal destruction via Nrf2-dependent antioxidant system

Periodontitis is a widespread oral disease characterized by continuous inflammation of the periodontal tissue and an irreversible alveolar bone loss, which eventually leads to tooth loss. Four-octyl itaconate(4-OI) is a cell-permeable itaconate derivative and has been recognized as a promising therapeutic target for the treatment of inflammatory diseases. Here, we explored, for the first time,the protective effect of 4-OI on inhibiting periodontal destruction, ameliorating local inflammation, and the underlying mechanism in periodontitis. Here we showed that 4-OI treatment ameliorates inflammation induced by lipopolysaccharide in the periodontal microenvironment. 4-OI can also significantly alleviate inflammation and alveolar bone loss via Nrf2 activation as observed on samples from experimental periodontitis in the C57BL/6 mice. This was further confirmed as silencing Nrf2 blocked the antioxidant effect of 4-OI by downregulating the expression of downstream antioxidant enzymes. Additionally, molecular docking simulation indicated the possible mechanism under Nrf2 activation. Also, in Nrf2-/-mice, 4-OI treatment did not protect against alveolar bone dysfunction due to induced periodontitis, which underlined the importance of the Nrf2 in 4-OI mediated periodontitis treatment.Our results indicated that 4-OI attenuates inflammation and oxidative stress via disassociation of KEAP1-Nrf2 and activation of Nrf2 signaling cascade. Taken together, local administration of 4-OI offers clinical potential to inhibit periodontal destruction,ameliorate local inflammation for more predictable periodontitis.

Thin Layer Chromatography and Methodological Investigation of Matrine in Zhukuqin Granule

[Objectives] To establish a sensitive and simple thin layer chromatography method of matrine in Zhukuqin granule. [Methods]With Zhukuqin granule as control medicinal materials,matrine as reference substance,we chose orthogonal test method,systematically investigated various factors affecting the thin layer chromatography,and selected the best thin layer chromatography conditions. [Results] We screened out the best thin layer chromatography of matrine in the Zhukuqin granule with chloroform as extraction solvent,solvent extraction and ultrasonic extraction as extraction method,toluene-acetone-ethyl acetate-ammonia solution( 2:3:4:0. 5) as developing agent,improvement of bismuth potassium iodide solution as color developing agent. [Conclusions] The separating degree of this method was high,it had good repeatability,and it could be used to control quality of Zhukuqin granule.

Whole-exome sequencing identifies ECPAS as a novel potentially pathogenic gene in multiple hereditary families with nonsyndromic orofacial cleft

Dear Editor,Orofacial cleft(OFC),which includes cleft lip and/or palate(CL/P)and cleft palate(CP),is the most common congenital craniofacial structural disorder,with a prevalence of 1.416%。among live infants worldwide(Massenburg et al.,2021).Nonsyndromic OFC(NSOFC),which does not contain other malformations as syndromic OFC(SOFC),accounts for 70%of cases and is believed to have complex etiologies.Notably,it has been established that genetic factors play a crucial role in the occurrence of NSOFC(Dixon et al.,2011).

Identification of rare PTCH1 nonsense variant causing orofacial cleft in a Chinese family and an up-to-date genotype- phenotype analysis

The Patched 1(PTCH1)gene encodes a membrane receptor involved in the Hedge-hog(Hh)signaling pathway,an abnormal state of which may result in congenital defects or hu-man tumors.In this study,we conducted whole-exome sequencing on a three-generation Chinese family characterized with variable penetrance of orofacial clefts.A rare heterozygous variant in the PTCH1 gene(c.2833C>T p.R945X)was identified as a disease-associated mutation.Structural modeling revealed a truncation starting from the middle of the second extracellular domain of PTCH1 protein.This may damage its ligand recognition and sterol transportation abilities,thereby affecting the Hh signaling pathway.Biochemical assays indi-cated that the R945X protein had reduced stability compared to the wild-type in vitro.In addi-tion,we reviewed the locations and mutation types of PTCH1 variants in individuals with clefting phenotypes,and analyzed the associations between clefts and locations or types of variants within PTCH1.Our findings provide further evidence that PTCH1 variants result in or-ofacial clefts,and contributed to genetic counseling and clinical surveillance in this family.

A novel FZD6 mutation revealed the cause of cleft lip and/or palate in a Chinese family

Cleft lip and/or palate(CL/P)is a most common craniofacial birth defect which has multifactorial etiology.In our study,we aimed to discover the underlying etiological gene variation in a Chinese family diagnosed as non-syndromic CL/P(NSCL/P).The blood sample of the proband and her parents were detected by whole exome sequencing.The Mendelian inheritance pattern,allele frequency,variation location,function analysis and literature search were applied to filtrate and screen the mutation.Besides,the candidates were confirmed by Sanger sequencing.We meanwhile explored the conservative analysis and protein homology simulation.As a result,a start-lost mutation c.1A>GAtg/Gtg in the Frizzled-6(FZD6)gene predicting p.Met1 was detected.The variation has not been reported before and was predicted to be harmful.The alteration caused missing of two starting amino acids that are evolutionarily conserved for FZD6 protein.Moreover,the specific structure of the mutant protein obviously changed according to the results of the homologous model.In conclusion,the results suggest c.1A>GAtg/Gtg in the FZD6(NM_001164616)might be the genetic etiology for non-syndromic CL/P in this pedigree.Furthermore,this finding provided new etiologic information,supplementing the evidence that FZD6 is a strong potential gene for CL/P.