Objective Tandem Mass Tag(TMT)in vitro quantitative proteomics technology was used to study the mechanism of Xiaozhongzhitong Mixture in the treatment of rat skin flap ischemia-reperfusion injury(FIRI).Methods Thirty SD rats were randomly divided into a normal group,a model group,and a Xiaozhongzhitong mixture group(Xiaozhong group),with 10 rats in each group.After intraperitoneal anesthesia,the model group and Xiaozhong group were completely freed with a pedicled skin flap of about 5 cm*6 cm in size on the back to simulate FIRI;the Xiaozhong group was given Xiaozhongzhitong Mixture(1.8 mL·kg-1)by gavage,once a day,the normal group and the model group were given normal saline(1.8 mL·kg-1)by gavage,once a day,and the rats in the three groups were continuously gavaged for 7 days;after 7 days,the dorsal skin flaps of the rats were collected for TMT proteomics analysis,screen out the differential proteins,perform Gene Ontology(GO)enrichment analysis,pathway enrichment(KEGG)analysis,common differential protein analysis,and establish Protein-Protein Interaction(PPI)network analysis.Results In this experiment,a total of 5005 proteins were screened by protein quantification,of which 4996 proteins were identified,and 141 up-regulated and 132 down-regulated differentially expressed proteins were screened out between the detumescence group/model group.GO enrichment analysis found that the differentially expressed proteins were mainly involved in molecular mechanisms such as organism cell metabolism,occurrence and combination of cellular components,catabolism,organelle formation,mitochondria formation,intracellular substance catalysis,molecular binding,hydrolase activation,and nucleotide regulation.KEGG pathway analysis found that the differentially expressed proteins were mainly involved in signaling pathways such as organismal cell metabolism,ribosome transcription and translation,proteasome system,and cell cycle.PPI analysis found that the differentially expressed co-expressed proteins Mrpl32,Psma2,Mrpl34,Psma4,Psmb1,and Srp9 were located at key nodes of the interaction network.Conclusion The treatment of FIRI with Xiaozhongzhitong Mixture may be related to the mitochondrial ribosome-related signaling pathway and the 20S proteasome system signaling pathway,among which the differential proteins Mrpl32,Psma2,Mrpl34,Psma4 and Psmb1 may be the key targets of treatment.

BACKGROUND:In recent years,it has been found that some traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of the skin flap,promote vascular regeneration of the skin flap and prevent skin flap necrosis by activating autophagy. OBJECTIVE:To review the research progress of traditional Chinese medicine monomer regulating autophagy in preventing flap necrosis. METHODS:The Chinese and English key words were"traditional Chinese medicine(TCM),autophagy,skin flaps".The first author searched the relevant articles published in CNKI and PubMed databases from January 2010 to October 2022.A total of 196 articles were retrieved in the preliminary screening and then screened according to the inclusion and exclusion criteria.The quality assessment was conducted by reading the literature titles and abstracts.Finally,55 articles were summarized. RESULTS AND CONCLUSION:(1)The regulation of autophagy is mediated by AMPK/mTOR,PI3K/AKT and other signaling pathways.Activation of autophagy can alleviate the oxidative stress and apoptosis of the flap,promote the regeneration of blood vessels in the flap,and prevent flap necrosis.(2)Terpenoids(Betulinic acid,Andrographolide,Notoginseng Triterpenes,Catalpa),phenolic compounds(Resveratrol,Curcumin,Gastrodin),phenolic acids(Salvianolic acid B)and steroid compounds(Pseudoginsenoside F11)in traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of skin flap by regulating related signaling pathways to activate autophagy,promote skin flap angiogenesis and promote skin flap survival.(3)Studying the research progress of traditional Chinese medicine monomer to prevent flap necrosis by regulating autophagy can provide a reference and theoretical basis for traditional Chinese medicine to prevent flap necrosis and promote flap healing in the clinic.

OBJECTIVEUsing simultaneous multi-gene mutation screening to investigate the new method molecular epidemiological basis of 225 patients with nonsyndromic hearing loss in Tianjin, and verifying the for simultaneous multi-gene mutation screening.

METHODSTwo hundred and twenty-five patients with severe non-syndromic deafness from Tianjin CDPF and Association of the Deaf were included in the study. The single nucleotide polymorphisms scan, (SNPscan) technique was used for screening the 115 spots mutations in three common deafness-related genes (GJB2, SLC26A4, mtDNA 12S rRNA) of patients with nonsyndromic hearing loss in Tianjin. We verified the results by Sanger sequencing.

RESULTSAmong the 225 patients, there were 111 cases of deafness caused by mutation (49.3%). Using this method, up to 50% of the patients in our study were identified to have hereditary HL caused by mutations in the three genes. 56 patients with the GJB2 mutations were detected (24.9%), including 30 cases of homozygous mutations (13.3%), 26 patients (11.6%) of compound heterozygous mutations, and 21 cases (9.33%) of single heterozygous mutations. 50 patients with the SLC26A4 mutations were detected (22.2%), including 22 cases of homozygous mutations(9.8%), 28 patients (12.4%) of compound heterozygous mutations, and 22 cases (9.8%) of single heterozygous mutations. mtDNA 12S rRNA A1555G mutation was detected in 5 patients (2.2%). mtDNA 12S rRNA 1494C>T mutation was not detected. We verified the results by Sanger sequencing. The accuracy of the sequencing results was 100%. The SNPscan cost eight hours and 160 yuan (each sample).

CONCLUSIONSApplying SNPscan technology can be accurate, rapid and cost-effective diagnostic screening in patients with hearing loss for etiology investigation. It is expected to become an effective means of large-scale genetic testing for hereditary deafness.

Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; methods ; DNA, Mitochondrial ; genetics ; Deafness ; genetics ; Genetic Testing ; methods ; Heterozygote ; Homozygote ; Humans ; Membrane Transport Proteins ; genetics ; Mutation ; Polymorphism, Single Nucleotide ; RNA, Ribosomal ; genetics