Objective:To investigate the transcriptomic signature of refractory nasal polyps (NP) after endoscopic sinus surgery.Methods:Tissue samples were collected from 36 patients with NP who underwent endoscopic sinus surgery at the Seventh Affiliated Hospital of Sun Yat-sen University and the First Affiliated Hospital of Sun Yat-sen University from January 2020 to December 2021. Raw sequencing data of normal nasal mucosa samples were downloaded from publicly available GEO database (Accession Number: GSE136825). Differential expression genes (DEGs) and Gene Ontology (GO) enrichment analysis were conducted to analyze the differences between refractory NP and normal controls, as well as among refractory, controlled, and partially controlled NP. Hierarchical clustering method was employed to analyze the inflammatory endotypes of NP. Weighted Gene Co-expression Network Analysis (WGCNA) and STRING database were used in combination with Cytoscape software to identify the characteristic transcriptional expression profiles of refractory NP. R software (version 4.3.1) was used for statistical analysis.Results:Refractory NP patients had significantly higher asthma comorbidity rates than controlled/partially controlled groups ( P<0.05). The numbers and percentages of peripheral blood and tissue eosinophilic granulocytes were significantly higher in the refractory subgroup than in the other two subgroups ( P<0.05). Compared to normal mucosa, controlled and partially controlled NP groups, 27 genes were consistently upregulated in refractory NP. Hierarchical cluster analysis showed that the refractory NP exhibited a mixed endotype dominated by type 2 inflammation with co-existing type 1 features. Differential genes were enriched in extracellular matrix organization, leukocyte activation, cytokine receptor activation, cystatin-mediated protease inhibition, granule exocytosis, and olfactory nerve development regulation. Further WGCNA analysis and protein-protein interaction network identified 33 hub genes represented by ITGAM, NCF1, NCF2, CD1C, PTAFR, CLEC10A, SIRPA, TREM2, ALOX5AP, PTGDR2 (officially PTGDR), F13A1, DUOX2, NOS2, CTSG, and SALL1.Conclusion:This study reveals the distinctive transcriptional signature of refractory NP through transcriptomic methods, providing novel research avenues and therapeutic targets for the treatment of refractory NP after surgery.

Objective:To investigate the transcriptomic signature of refractory nasal polyps (NP) after endoscopic sinus surgery.Methods:Tissue samples were collected from 36 patients with NP who underwent endoscopic sinus surgery at the Seventh Affiliated Hospital of Sun Yat-sen University and the First Affiliated Hospital of Sun Yat-sen University from January 2020 to December 2021. Raw sequencing data of normal nasal mucosa samples were downloaded from publicly available GEO database (Accession Number: GSE136825). Differential expression genes (DEGs) and Gene Ontology (GO) enrichment analysis were conducted to analyze the differences between refractory NP and normal controls, as well as among refractory, controlled, and partially controlled NP. Hierarchical clustering method was employed to analyze the inflammatory endotypes of NP. Weighted Gene Co-expression Network Analysis (WGCNA) and STRING database were used in combination with Cytoscape software to identify the characteristic transcriptional expression profiles of refractory NP. R software (version 4.3.1) was used for statistical analysis.Results:Refractory NP patients had significantly higher asthma comorbidity rates than controlled/partially controlled groups ( P<0.05). The numbers and percentages of peripheral blood and tissue eosinophilic granulocytes were significantly higher in the refractory subgroup than in the other two subgroups ( P<0.05). Compared to normal mucosa, controlled and partially controlled NP groups, 27 genes were consistently upregulated in refractory NP. Hierarchical cluster analysis showed that the refractory NP exhibited a mixed endotype dominated by type 2 inflammation with co-existing type 1 features. Differential genes were enriched in extracellular matrix organization, leukocyte activation, cytokine receptor activation, cystatin-mediated protease inhibition, granule exocytosis, and olfactory nerve development regulation. Further WGCNA analysis and protein-protein interaction network identified 33 hub genes represented by ITGAM, NCF1, NCF2, CD1C, PTAFR, CLEC10A, SIRPA, TREM2, ALOX5AP, PTGDR2 (officially PTGDR), F13A1, DUOX2, NOS2, CTSG, and SALL1.Conclusion:This study reveals the distinctive transcriptional signature of refractory NP through transcriptomic methods, providing novel research avenues and therapeutic targets for the treatment of refractory NP after surgery.

ObjectiveThe lung mesenchymal stem cells （LMSCs） induced by D-galactose （D-gal） were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 （NAMPT/SIRT1） signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation， and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions （5%， 10%， 20%， 40%， and 80%） of Wenfei Huaxian decoction-containing serum for 24 h， and the cell proliferation level was detected by methyl thiazolyl tetrazolium （MTT） method. The cells were randomly divided into blank serum group， model group， and high， medium， and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase （SA-β-gal） staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors （p16 and p53）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group， the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h （P<0.01）. Compared with the model group， the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum （P<0.05， P<0.01）. Compared with the blank serum group， the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced， and the content of NAD⁺ was increased （P<0.01）. The expression levels of senescence factors p16 and p53， as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant， were significantly increased （P<0.01）. The expression of senescence-associated proteins p16， p21， and p53 increased （P<0.01）， and the protein expression of NAMPT， SIRT1， peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α）， and forkhead box family transcription factor O1 （FoxO1） decreased （P<0.01）. Compared with the model group， the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced， and the content of NAD⁺ was decreased （P<0.01）. The senescence factors （p16 and p53） and inflammatory factors （IL-6 and TNF-α） in the cell culture supernatant were significantly decreased （P<0.01）. The expression of senescence-associated proteins （P16， P21， and P53） decreased （P<0.05， P<0.01）. The protein expressions of NAMPT， SIRT1， PGC-1α， and FoxO1 were significantly up-regulated （P<0.05， P<0.01）. ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs， and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.

Objective:To explore the relieving effect of adhesive Bifidobacterium on constipation and its effect on intestinal flo-ra and intestinal morphology in mice.Methods:Detection of adhesion of 14 strains of Bifidobacterium.Fifty mice were randomly divid-ed into control group,model group,positive drug group,low adhesion group and high adhesion group,with 10 mice in each group.The control group and model group received 0.2 ml sterile double distilled water,the positive drug group received 0.2 ml 1.52 mg/kg lactulose solution,and the low and high adhesion groups received 0.2 ml 109 CFU/ml corresponding to Bifidobacterium solution for 37 days.The constipation model was established from the 31st day.The other 4 groups except the control group were given 0.2 ml 10 mg/(kg·bw)loperamide hydrochloride aseptic aqueous solution for 7 days.After the establishment of the model,each mouse was fed with 0.2 ml activated carbon suspension,and the excretion time of the first black stool was measured.After fasting for 12 hours,the mice were given aseptic double steamed water or loperamide hydrochloride,and 1 hour later,0.2 ml activated carbon suspension was given to the stomach.After 30 min,all the mice were killed.The distance between the intestinal length and the intestinal propul-sion of the activated carbon suspension was measured,and the intestinal propulsion rate was calculated.The fresh feces of mice on the 30th and 36th day were collected within 4 hours,and the fecal moisture content and short chain fatty acid(SCFAs)content were deter-mined.Observation of histology of small intestine by HE staining.The expression of zonula occluden-1(ZO-1)and Occludin was detected by immunofluorescence.Detection of aquaporin,c-kit and serotonin related gene transcription by qRT-PCR.The levels of 5-hydroxytryptamine(5-HT),IL-1β and TNF-α were detected by ELISA.The relative content of intestinal microflora was detected by 16S rRNA high-throughput sequencing.Results:CCFM 622,CCFM 624,CCFM 626,CCFM 641 and CCFM 642 were selected as the bacteria group used in the low adhesion group,and CCFM 16,CCFM 625,CCFM 643,CCFM 666 and CCFM 668 as the bacteria group used in the high adhesion group.Compared with control group,the first black stool excretion time,AQP4,AQP8 mRNA,5-HT,IL-1β and TNF-α levels in the model group,positive drug group,low adhesion group and high adhesion group increased(P<0.05),while the intestinal propulsion rate,fecal water content,ZO-1,Occludin fluorescence intensity,c-kit,5-HT4R,Tph1 mRNA,acetic acid,propionic acid and butyric acid decreased(P<0.05).Compared with positive drug group and high adhesion group,the first black stool excretion time,AQP4,AQP8 mRNA,5-HT,IL-1β and TNF-α in low adhesion group increased(P<0.05),while intestinal pro-pulsion rate,fecal water content,ZO-1,Occludin fluorescence intensity,c-kit,5-HT4R,Tph1 mRNA,acetic acid,propionic acid and butyric acid decreased in low adhesion group(P<0.05).In the control group,the mucosal structure was intact,the small intesti-nal villi were neat and uniform,there was no injury or wrinkle,and the cell structure was intact;in model group,the intestinal villi were seriously ruptured and atrophied,and the goblet cells were incomplete;compared with model group,the morphology of small intestinal villi in positive drug group,low adhesion group and high adhesion group were improved,and the number of goblet cells increased,and the improvement in positive drug group and high adhesion group were more obvious than that in low adhesion group.Compared with control group,the Shannon index and Firmicutes/Bacteroidetes(F to B)ratio in model group decreased(P<0.05),and there were differences in genus level(P<0.05).Compared with model group,Shannon index and F to B ratio increased in positive drug group,low adhesion group and high adhesion group(P<0.05).The sample points in the model group could be distinguished from the control group,positive drug group,low adhesion group and high adhesion group,but the coincidence degree of sample points in control group,positive drug group,low adhesion group and high adhesion group were higher.Conclusion:Bifidobacterium can regu-late intestinal flora of constipation mice,improve intestinal running efficiency and protect intestinal mucus barrier.Bifidobacterium with high adhesion is better than Bifidobacterium with low adhesion in relieving constipation in mice.

Objective:To explore the role and relationship between oxidative stress and immune infiltration in idiopathic pul-monary fibrosis(IPF),and to predict the relevant therapeutic herbal medicines and active ingredients.Methods:GSE10667 gene expression profiles were downloaded from GEO database to obtain differential expression genes,differential expression of oxidative stress genes(DEOSGs)were identified in combination with oxidative stress genes.GSEA was used to evaluate the pathways and biologi-cal processes in IPF,and GO,KEGG and PPI network analysis were performed on DEOSGs.Candidate central genes were derived from PPI results and CytoHubba,and GSE110147 was validated as an independent group to identify central genes;in addition,the immune microenvironment of samples was evaluated using CIBERSORTF,and correlation between central gene levels and relative proportion of immune cells was explored;finally,therapeutic herbal medicines and components were predicted by central genes,and mole-cular docking verification was carried out.Results:A total of 51 DEOSGs,four central genes(ICAM-1,APOE,MMP-1,TGF-β2)were obtained;DEOSGs were mainly related to oxidative stress,immune response,etc;four central gene levels were closely correlated with 8 relative proportions of immune cells;therapeutic herbal medicines included 4 flavors such as Huangqi and Chuanxiong,and the active ingredients included 8 kinds of β-carotene,etc,the molecular docking results were stable.Conclusion:Oxidative stress and immune firing are exist in IPF,and oxidative stress may be recognized by immune cells or directly activate immune cells.

Esophageal stent implantation is the most commonly used clinical treatment for malignant esophageal obstruction.Driven by several biotechnological advances,great progress has been made in the clinical application of esophageal stents in the past few years.Therefore,it is extremely important for interventional physicians to have a thorough understanding of the mechanical properties and material properties of various esophageal stents.More and more clinical evidences indicate that esophageal stents,especially 125I radioactive esophageal stent,possess exceptional targeted drug delivery capabilities,which enables the physicians to formulated an individualized diagnosis and treatment plan that is more suitable for the patient's situation,so as to significantly improve the therapeutic accuracy.This review aims to systematically introduce the respective advantages of different esophageal stents,to discuss the biggest problems faced in the current clinical work,and to summarize the latest advances in cancer treatment,including the fields of both basic experimental study and clinical research.This review also explicates the design ideas and working principles of various esophageal stents in detail,discusses the development prospect of stent surface modification technology such as covered stents and drug-eluting stents,and makes judgment of the effectiveness and safety of these types of stents in alleviating malignant esophageal obstruction.

Objective:To investigate the trend of postoperative cavity status in patients with eosinophilic chronic sinusitis with nasal polyps (eCRSwNP) who underwent total nasalization surgery and partial reboot surgery. And to discuss the relationship between tissue eosinophil counts and status of postoperative cavity.Methods:Patients with eCRSwNP in four tertiary medical centers (Longgang ENT Hospital, Xiamen Humanity Hospital, Guangdong Clifford Hospital and the First Affiliated Hospital of Sun Yat-Sen University) from March 2018 to October 2021 were divided into 2 groups. The group without previous surgery history was performed for the nasalization surgery, and another group with previous surgery history underwent the part-reboot surgery. The follow-up time after operation was defined as the following 5 stages: 6, 12, 20-24, 36 and more than 42 months. According to FESS-95 Guangzhou standard, status of sinus cavity was assessed and classified into 3 categories: good, better and bad. The association between the sinus cavity status and tissue eosinophil counts in the above 5 stages was analyzed by one-way ANOVA, and P<0.05 was considered statistically significant. Results:A total of 72 eCRSwNP patients finished the follow-up in this study. There were 47 males and 25 females in these patients, aged from 11 to 67 years. A total of 50 cases underwent nasalization surgery and 22 cases underwent partial reboot surgery. With the follow-up time from 6 to 48 months, there were 72 cases (100.0%) who completed 6 months and 12 months follow up, 46 cases (63.9%) for 20-24 months, 36 cases (50.0%) for 32-36 months and 16 cases (22.2%) with the follow-up time more than 42 months. No matter what kind of surgery, there was no "bad" situation of the surgical cavity status 6 months after the operation, and the differentiation gradually occurred more than 12 months after the surgery. Moreover, the rates of "good" cavity status for the 5 stages in the group of nasalization surgery were 78.0%, 66.0%, 56.7%, 47.6% and 42.9%, and were 63.6%, 45.5%, 25.0%, 20.0% and 11.1% in the partial reboot surgery group, respectively, suggesting that the status of nasal cavity in nasalization surgery group was always better than that in partial reboot surgery group in every period. In addition, the "bad" rate was 0, 8.0%, 10.0%, 14.3% and 28.6% in the group of nasalization surgery, and was 0, 27.3%, 18.8%, 33.3% and 55.6% in the partial reboot surgery group, respectively. The average percentage of tissue eosinophil counts in the 72 cases was 42.1%, which had no obvious effect on the status of the surgical cavity ( P>0.05). Conclusions:For eCRSwNP patients, the operative cavity status in the patients without previous operation history treated with nasalization surgery is good. The time of 1-2 years after surgery is the main period for sinus lesions. The counts of tissue eosinophils has no significant influence on surgical sinus cavity status in the eCRSwNP patients.

Mitochondrial dynamics, the processes of mitochondrial fusion and fission maintain homeostasis, are precisely regulated by fusion/fission-related proteins, and play an important physiological role in mitochondrial metabolism, quality and function. The aberrant changes of these proteins can trigger mitochondrial dynamics imbalance, which cause mitochondrial dysfunctions and result various disease states. This article focuses on gene knockout technology, and reviews the role and application progress of genes encoding for fusion and fission knockout mice in insulin resistance researches, in order to lay a foundation for future studies on signal transduction mechanism of mitochondrial dynamics imbalance in insulin resistance.
Animals ; Gene Knockout Techniques ; Insulin Resistance ; Mice ; Mitochondria ; Mitochondrial Dynamics ; Mitochondrial Proteins

Primary biliary cholangitis (PBC) is a chronic cholestatic autoimmune disease and may progress to liver fibrosis, liver cirrhosis, and even end-stage liver disease without timely and effective treatment. The main purpose of PBC treatment is to prevent the development of end-stage liver disease and improve related clinical symptoms and quality of life. Timely and accurate risk stratification may bring benefits to patients with refractory PBC, and an accurate evaluation of the prognosis of PBC patients is of vial importance for clinical treatment. This article reviews the research advances in the prognostic factors for PBC from the aspects of host factors, laboratory examination, and pharmacotherapy.