AIM:To explore the potential mecha-nism of arecoline in promoting oral submucous fi-brosis based on key factors of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway.METHODS:SD rats were randomly divided into arecoline low-dose group,arecoline medium-dose group,and are-coline high-dose group(5,10,and 15 mg/mL).The oral buccal mucosa was injected with the corre-sponding concentration of arecoline(ANE)solution to induce the establishment of oral submucous fi-brosis(OSF)models,with 8 rats in each group.An-other 8 unmodeled rats were selected as the blank group,and the changes in mouth opening of the rats were detected after 8 weeks of grouping and intervention.HE and Masson staining were used to observe the pathological changes of oral buccal mucosa,measure the length of epithelial staple process and calculate collagen volume fraction.Western blot and qRT-PCR were used to detect the expression of collagen-Ⅰ(COL-Ⅰ),E-cadherin,fibro-nectin(FN)and PI3K/Akt/mTOR signaling pathway-related proteins and mRNA in rat oral buccal muco-sa.The levels of tumor necrosis factor(TNF)-α,in-terleukin(IL)-1β and transforming growth factor(TGF)-β1 in rat serum were detected by ELISA.Hu-man immortalized keratinocytes(HaCaT cell line)were cultured in vitro,and the effects of different concentrations of arecoline,PI3K activator,and PI3K inhibitor on the survival rate of HaCaT cells were investigated by CCK-8 method.According to the results of CCK-8,the concentration of arecoline 75 μg/mL,the concentration of PI3K activator 10μmol/L,and the concentration of PI3K inhibitor 2μmol/L were selected as the subsequent experi-mental concentrations.The cells were set as blank group,arecoline group,PI3K activator group,PI3K inhibitor group,and arecoline+PI3K inhibitor group.The mRNA expression levels of COL-Ⅰ,E-cad-herin,FN,PI3K,Akt,and mTOR in each group of cells were detected by qRT-PCR.The levels of TNF-α,IL-1β and TGF-β1 in each group of cells were de-tected by ELISA.RESULTS:Compared with the blank group,the arecoline low-dose group,the are-coline medium-dose group,and the arecoline high-dose group significantly reduced the mouth open-ing,significantly shortened the length of the epi-thelial staple process,significantly increased the collagen volume fraction,inflammatory cell infiltra-tion,and severe pathological damage.The protein expression levels of COL-Ⅰ,FN,p-PI3K,p-Akt,and p-mTOR were up-regulated,and the protein expres-sion levels of E-cadherin were down-regulated.The mRNA expressions of COL-Ⅰ,FN,PI3K,Akt,and mTOR were significantly increased.The mRNA ex-pression of E-cadherin was significantly reduced,and the levels of TNF-α,IL-1β,and TGF-β1 were sig-nificantly increased(all P＜0.05 or P＜0.01).Com-pared with the blank group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the arecoline group and the PI3K activator group were up-regulated,and the mRNA expression lev-els of E-cadherin were down-regulated.Compared with the blank group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the PI3K inhibitor group were down-regulated,and the mRNA expression levels of E-cadherin were up-regulated.Compared with the PI3K inhibitor group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the arecoline+PI3K inhibi-tor group were up-regulated,the mRNA expression levels of E-cadherin were down-regulated,and the levels of TNF-α,IL-1β,and TGF-β1 were significant-ly increased(all P＜0.05 or P＜0.01).CONCLUSION:Arecoline can significantly promote oral submucous fibrosis,which may play a pro-fibrotic role by up-regulating the PI3K/Akt/mTOR signaling pathway.

Objective To establish a rapid detection method for Clostridium botulinum in food.Methods A rapid detection method based on RPA-CRISPR/Cas12a was developed by integrating recombinase polymerase amplification(RPA)with the CRISPR-Cas12a system.After the reaction conditions were optimized,the method's sensitivity,specificity,and usefulness were methodically confirmed.Results and Conclusion The optimized method achieved detection within 1 hour,with a limit of detection(LOD)of 1.91 copies/μL.No cross-reactivity was observed with non-target pathogens.The RPA-CRISPR/Cas12a-based detection method developed in this study exhibits high specificity,sensitivity,and operational simplicity and may provide a feasible solution for the rapid detection of foodborne pathogens.

BACKGROUND: Disease-modifying therapies have been approved for the treatment of relapsing multiple sclerosis (RMS). The present study aims to examine the safety of teriflunomide in Chinese patients with RMS. METHODS: This non-randomized, multi-center, 24-week, prospective study enrolled RMS patients with variant (c.421C>A) or wild type ABCG2 who received once-daily oral teriflunomide 14 mg. The primary endpoint was the relationship between ABCG2 polymorphisms and teriflunomide exposure over 24 weeks. Safety was assessed over the 24-week treatment with teriflunomide. RESULTS: Eighty-two patients were assigned to variant ( n  = 42) and wild type groups ( n  = 40), respectively. Geometric mean and geometric standard deviation (SD) of pre-dose concentration (variant, 54.9 [38.0] μg/mL; wild type, 49.1 [32.0] μg/mL) and area under plasma concentration-time curve over a dosing interval (AUC tau ) (variant, 1731.3 [769.0] μg∙h/mL; wild type, 1564.5 [1053.0] μg∙h/mL) values at steady state were approximately similar between the two groups. Safety profile was similar and well tolerated across variant and wild type groups in terms of rates of treatment emergent adverse events (TEAE), treatment-related TEAE, grade ≥3 TEAE, and serious adverse events (AEs). No new specific safety concerns or deaths were reported in the study. CONCLUSION: ABCG2 polymorphisms did not affect the steady-state exposure of teriflunomide, suggesting a similar efficacy and safety profile between variant and wild type RMS patients. REGISTRATION NCT04410965, https://clinicaltrials.gov .
Humans ; Crotonates/adverse effects* ; Toluidines/adverse effects* ; Nitriles ; Hydroxybutyrates ; Female ; Male ; Adult ; ATP Binding Cassette Transporter, Subfamily G, Member 2/genetics* ; Middle Aged ; Multiple Sclerosis, Relapsing-Remitting/genetics* ; Prospective Studies ; Young Adult ; Neoplasm Proteins/genetics* ; East Asian People

BACKGROUND: G protein-coupled receptor kinase 2 (GRK2) could participate in the regulation of diverse cells via interacting with non-G-protein-coupled receptors. In the present work, we explored how paroxetine, a GRK2 inhibitor, modulates the differentiation and activation of immune cells in rheumatoid arthritis (RA). METHODS: The blood samples of healthy individuals and RA patients were collected between July 2021 and March 2022 from the First Affiliated Hospital of Anhui Medical University. C57BL/6 mice were used to induce the collagen-induced arthritis (CIA) model. Flow cytometry analysis was used to characterize the differentiation and function of dendritic cells (DCs)/T cells. Co-immunoprecipitation was used to explore the specific molecular mechanism. RESULTS: In patients with RA, high expression of GRK2 in peripheral blood lymphocytes, accompanied by the increases of phosphatidylinositol 3 kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR). In animal model, a decrease in regulatory T cells (T regs ), an increase in the cluster of differentiation 8 positive (CD8 + ) T cells, and maturation of DCs were observed. Paroxetine, when used in vitro and in CIA mice, restrained the maturation of DCs and the differentiation of CD8 + T cells, and induced the proportion of T regs . Paroxetine inhibited the secretion of pro-inflammatory cytokines, the expression of C-C motif chemokine receptor 7 in DCs and T cells. Simultaneously, paroxetine upregulated the expression of programmed death ligand 1, and anti-inflammatory cytokines. Additionally, paroxetine inhibited the PI3K-AKT-mTOR metabolic pathway in both DCs and T cells. This was associated with a reduction in mitochondrial membrane potential and changes in the utilization of glucose and lipids, particularly in DCs. Paroxetine reversed PI3K-AKT pathway activation induced by 740 Y-P (a PI3K agonist) through inhibiting the interaction between GRK2 and PI3K in DCs and T cells. CONCLUSION Paroxetine exerts an immunosuppressive effect by targeting GRK2, which subsequently inhibits the metabolism-related PI3K-AKT-mTOR pathway of DCs and T cells in RA.
G-Protein-Coupled Receptor Kinase 2/metabolism* ; Arthritis, Rheumatoid/immunology* ; Animals ; Dendritic Cells/metabolism* ; Paroxetine/therapeutic use* ; Proto-Oncogene Proteins c-akt/metabolism* ; Mice ; Humans ; Mice, Inbred C57BL ; Signal Transduction/drug effects* ; Male ; Phosphatidylinositol 3-Kinases/metabolism* ; Lymphocyte Activation/drug effects* ; Female ; T-Lymphocytes/metabolism* ; Middle Aged

Objective To establish a combined model of spectral CT multi-parameters and clinical features to distinguish between gastric poorly cohesive carcinoma and tubular adenocarcinoma.Methods A total of 87 patients with gastric cancer confirmed by postoperative pathology were retrospectively selected,including 26 patients with poorly cohesive carcinoma and 61 patients with tubular adenocarcinoma.Predictors were identified by univariate and multivariate logistic regression analyses,and a combined model was established.The area under the curve(AUC)of receiver operating characteristic(ROC)curve was used to evaluate the differential diagnostic efficiency of the parameters and the model.The AUC was compared by DeLong method.Results The gender[odds ratio(OR)5.124,P=0.004],normalized iodine density in the arterial phase(nIoDAP)(OR 5.789,P=0.017),arterial enhancement fraction(AEF)(OR 7.007,P=0.002)and ΔIoD(OR 0.025,P=0.021)were identified as independent predictors for poorly cohesive carcinoma by logistic regression analysis.The AUC of combined model established by four variables in distinguishing poorly cohesive carcinoma and tubular adenocarcinoma was 0.837[95％confidence interval(CI)0.716-0.907],which was significantly higher than that of single tumor spectral CT parameters(P＜0.01).Conclusion The combined model based on patients'gender and tumor spectral CT parameters(nIoDAP,AEF and ΔIoD)can effectively distinguish gastric poorly cohesive carcinoma and tubular adenocarcinoma,providing a basis for gastric cancer patients'individualized treatment strategy.

AIM:To explore the potential mecha-nism of arecoline in promoting oral submucous fi-brosis based on key factors of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway.METHODS:SD rats were randomly divided into arecoline low-dose group,arecoline medium-dose group,and are-coline high-dose group(5,10,and 15 mg/mL).The oral buccal mucosa was injected with the corre-sponding concentration of arecoline(ANE)solution to induce the establishment of oral submucous fi-brosis(OSF)models,with 8 rats in each group.An-other 8 unmodeled rats were selected as the blank group,and the changes in mouth opening of the rats were detected after 8 weeks of grouping and intervention.HE and Masson staining were used to observe the pathological changes of oral buccal mucosa,measure the length of epithelial staple process and calculate collagen volume fraction.Western blot and qRT-PCR were used to detect the expression of collagen-Ⅰ(COL-Ⅰ),E-cadherin,fibro-nectin(FN)and PI3K/Akt/mTOR signaling pathway-related proteins and mRNA in rat oral buccal muco-sa.The levels of tumor necrosis factor(TNF)-α,in-terleukin(IL)-1β and transforming growth factor(TGF)-β1 in rat serum were detected by ELISA.Hu-man immortalized keratinocytes(HaCaT cell line)were cultured in vitro,and the effects of different concentrations of arecoline,PI3K activator,and PI3K inhibitor on the survival rate of HaCaT cells were investigated by CCK-8 method.According to the results of CCK-8,the concentration of arecoline 75 μg/mL,the concentration of PI3K activator 10μmol/L,and the concentration of PI3K inhibitor 2μmol/L were selected as the subsequent experi-mental concentrations.The cells were set as blank group,arecoline group,PI3K activator group,PI3K inhibitor group,and arecoline+PI3K inhibitor group.The mRNA expression levels of COL-Ⅰ,E-cad-herin,FN,PI3K,Akt,and mTOR in each group of cells were detected by qRT-PCR.The levels of TNF-α,IL-1β and TGF-β1 in each group of cells were de-tected by ELISA.RESULTS:Compared with the blank group,the arecoline low-dose group,the are-coline medium-dose group,and the arecoline high-dose group significantly reduced the mouth open-ing,significantly shortened the length of the epi-thelial staple process,significantly increased the collagen volume fraction,inflammatory cell infiltra-tion,and severe pathological damage.The protein expression levels of COL-Ⅰ,FN,p-PI3K,p-Akt,and p-mTOR were up-regulated,and the protein expres-sion levels of E-cadherin were down-regulated.The mRNA expressions of COL-Ⅰ,FN,PI3K,Akt,and mTOR were significantly increased.The mRNA ex-pression of E-cadherin was significantly reduced,and the levels of TNF-α,IL-1β,and TGF-β1 were sig-nificantly increased(all P＜0.05 or P＜0.01).Com-pared with the blank group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the arecoline group and the PI3K activator group were up-regulated,and the mRNA expression lev-els of E-cadherin were down-regulated.Compared with the blank group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the PI3K inhibitor group were down-regulated,and the mRNA expression levels of E-cadherin were up-regulated.Compared with the PI3K inhibitor group,the mRNA expression levels of COL-Ⅰ,FN,PI3K,Akt,and mTOR in the cells of the arecoline+PI3K inhibi-tor group were up-regulated,the mRNA expression levels of E-cadherin were down-regulated,and the levels of TNF-α,IL-1β,and TGF-β1 were significant-ly increased(all P＜0.05 or P＜0.01).CONCLUSION:Arecoline can significantly promote oral submucous fibrosis,which may play a pro-fibrotic role by up-regulating the PI3K/Akt/mTOR signaling pathway.

Objective To establish a combined model of spectral CT multi-parameters and clinical features to distinguish between gastric poorly cohesive carcinoma and tubular adenocarcinoma.Methods A total of 87 patients with gastric cancer confirmed by postoperative pathology were retrospectively selected,including 26 patients with poorly cohesive carcinoma and 61 patients with tubular adenocarcinoma.Predictors were identified by univariate and multivariate logistic regression analyses,and a combined model was established.The area under the curve(AUC)of receiver operating characteristic(ROC)curve was used to evaluate the differential diagnostic efficiency of the parameters and the model.The AUC was compared by DeLong method.Results The gender[odds ratio(OR)5.124,P=0.004],normalized iodine density in the arterial phase(nIoDAP)(OR 5.789,P=0.017),arterial enhancement fraction(AEF)(OR 7.007,P=0.002)and ΔIoD(OR 0.025,P=0.021)were identified as independent predictors for poorly cohesive carcinoma by logistic regression analysis.The AUC of combined model established by four variables in distinguishing poorly cohesive carcinoma and tubular adenocarcinoma was 0.837[95％confidence interval(CI)0.716-0.907],which was significantly higher than that of single tumor spectral CT parameters(P＜0.01).Conclusion The combined model based on patients'gender and tumor spectral CT parameters(nIoDAP,AEF and ΔIoD)can effectively distinguish gastric poorly cohesive carcinoma and tubular adenocarcinoma,providing a basis for gastric cancer patients'individualized treatment strategy.

Objective This study aimed to clarify the intervention effect of salidroside(SAL)on lung injury caused by PM2.5 in mice and illuminate the function of SIRT1-PGC-1ɑ axis. Methods Specific pathogen-free(SPF)grade male C57BL/6 mice were randomly assigned to the following groups:control group,SAL group,PM2.5 group,SAL+PM2.5 group.On the first day,SAL was given by gavage,and on the second day,PM2.5 suspension was given by intratracheal instillation.The whole experiment consist of a total of 10 cycles,lasting 20 days.At the end of treatment,blood samples and lung tissues were collected and analyzed.Observation of pathological changes in lung tissue using inverted microscopy and transmission electron microscopy.The expression of inflammatory,antioxidants,apoptosis,and SIRT1-PGC-1ɑ proteins were detected by Western blotting. Results Exposure to PM2.5 leads to obvious morphological and pathologica changes in the lung of mice.PM2.5 caused a decline in levels of antioxidant-related enzymes and protein expressions of HO-1,Nrf2,SOD2,SIRT1 and PGC-1ɑ,and an increase in the protein expressions of IL-6,IL-1β,Bax,caspase-9 and cleaved caspase-3.However,SAL reversed the aforementioned changes caused by PM2.5 by activating the SIRT1-PGC-1α pathway. Conclusion SAL can activate SIRT1-PGC-1ɑ to ameliorate PM2.5-induced lung injury.

[Objective]To explore if the enhanced recovery after surgery (ERAS) protocol for pregnant women with gestational diabetes mellitus (GDM) who are undergoing elective cesarean delivery could cause perioperative glycemic abnormalities and heighten the risk of neonatal hypoglycemia.[Methods]A retrospective analysis was conducted on a cohort of pregnant women with singleton pregnancies who underwent elective cesarean sections and received ERAS between May 1,2022,and October 31,2023,at the Third Affiliated Hospital of Sun Yat-sen University. A total of 150 patients were included in this study,comprising the GDM group (n=75) and the non-GDM group (n=75). The study included pregnant women with good glycemic control (GDM) and maternal age (18-30 years;30-35 years;35-40 years;>40 years),BMI (<18.5 kg/m2;18.5-24.9 kg/m2;25-30 kg/m2;>30 kg/m2),and gestational age (within 7 days). We used these criteria to match 1∶1 non-GDM women as the control group. After administering preoperative oral carbohydrates,we observed the trends of maternal glycemic changes,including hyperglycemia and hypoglycemia,at any time of the day. We also evaluated the incidence of hypoglycemic low Apgar scores in newborns,abnormal pH values in blood gases,and the rate of transfer to the pediatric unit immediately after delivery.[Results]No significant difference was observed in fasting blood glucose levels on the day of surgery between the two groups of pregnant women[(4.4±0.5) mmol/L vs. (4.3±0.5) mmol/L,t=1.395,P=0.165]. The blood glucose peak was reached 30 minutes after consuming 300 mL (42.6 g of low-dose carbohydrate) of a light drink[(7.2±0.9) mmol/L vs. (6.4±0.8) mmol/L,t=5.773,P<0.001],with a subsequent decline in blood glucose levels. At the 120-minute mark,blood glucose had returned to the pre-oral carbohydrate level. The blood glucose levels in GDM groups was significantly higher than those in the non-GDM group (P<0.005). Although the incidence of hyperglycemia was significantly higher in the GDM group than in the non-GDM group at the 30-minute peak blood glucose level after oral carbohydrate intake,and the difference was statistically significant (17.3％ vs. 1.3％,x2=11.354,P<0.001),severe hyperglycemia (≥10 mmol/L) did not occur. The incidence of hypoglycemia was not significantly higher in neonates in the GDM group than in the non-GDM group (22.7％ vs. 28％,x2=0.564,P=0.453). The incidence of neonatal hypoglycemia in the GDM group was not significantly elevated in comparison to the non-GDM group after adjusting for age and BMI (Model 1),primiparity and gestational week of delivery (Model 2),hypertensive disorders of pregnancy (Model 3),cesarean section indications,time of cesarean section,and intraoperative hemorrhage (Model 4),and neonatal weight (Model 5).[Conclusion]In GDM patients with excellent glycemic control,an ERAS regimen with a low oral dose of carbohydrates prior to elective cesarean section does not increase the risk of preoperative serious hyperglycemia in mothers,nor does it increase the incidence of neonatal hypoglycemia.

Butyric acid is a type of short chain fatty acid and an important nutrient in intestinal epithelial cells.In addition to its important role in intestinal health,it has application value in anti-tumor,treatment of neuritis and di-abetes.At the same time,based on the demand for green development in the livestock industry,its anti-inflammato-ry effect can avoid the abuse of antimicrobial agents.As a green,pollution-free,and residue-free new feed,butyric acid ensures the sustainable and healthy development of the livestock industry.This article mainly summarizes the production of butyric acid in the intestine,its effects on the balance of gut microbiota,digestion ability,and inflam-mation in humans and animals,elaborates its application in human health and animal production.