G protein-coupled receptor kinase 2 (GRK2) participates in the phosphorylation and desensitization of G protein-coupled receptor (GPCR), impacting various biological processes such as inflammation and cell proliferation. Dysregulated expression and activity of GRK2 have been reported in multiple cells in rheumatoid arthritis (RA). However, whether and how GRK2 regulates synovial hyperplasia and fibroblast-like synoviocytes (FLSs) proliferation is poorly understood. In this study, we investigated the regulation of GRK2 and its biological function in RA. We found that GRK2 transmembrane activity was increased in FLSs of RA patients and collagen-induced arthritis (CIA) rats. Additionally, we noted a positive correlation between high GRK2 expression on the cell membrane and serological markers associated with RA and CIA. Immunoprecipitation-mass spectrometry and pull-down analyses revealed tumor necrosis factor receptor-associated factor 2 (TRAF2) as a novel substrate of GRK2. Furthermore, surface plasmon resonance (SPR) and molecular docking assays determined that the C-terminus of GRK2 binds to the C-terminus of TRAF2 at the Gln340 residue. GRK2 knockdown and the GRK2 inhibitor CP-25 attenuated synovial hyperplasia and FLS proliferation in CIA both in vitro and in vivo by decreasing GRK2 membrane expression and activity. Mechanistically, increased GRK2 transmembrane activity contributed to the recruitment of TRAF2 on the cell membrane, promoting GRK2-TRAF2 interactions that facilitate the recruitment of the E3 ubiquitin ligase TRIM47 to TRAF2. This enhanced TRAF2 Lys63 polyubiquitylation and induced nuclear factor (NF)-κB activation, leading to synovial hyperplasia and abnormal proliferation of FLSs. Our study provides a mechanistic and preclinical rationale for further evaluation of GRK2 as a therapeutic target for RA.

Objective To explore the effects of different daily doses of methadone maintenance treatment(MMT)on subjective craving and brain function under drug cues among heroin addicts,providing an objective basis for the formulation of methadone medi-cation plan in clinical practice.Methods Twenty-nine heroin addicts were included and grouped according to the daily dose of metha-done 40 mg,≤40 mg/d group 15 participants(group A),>40 mg/d group 14 participants(group B).The functional magnetic reso-nance imaging(fMRI)data of brain response induced by drug cues were collected using a 3.0T MR scanner,and the craving data induced by drug cues in the subjects were collected.Brain activity and behavioral data processing were analyzed using SPM8 and SPSS 20.0 software.Results There were no statistically significant differences in craving scores between the two groups of subjects exposed to drug cues(t=-0.69,P>0.05).The brain regions with differences in brain response included the left caudate nucleus(group A showed signifi-cantly enhanced response),and the strength of its response was negatively correlated with the daily dose of methadone(r=-0.465,P=0.025).Conclusion Higher doses of MMT may help control the value cognitive processing of drug cues in heroin addicts through the caudate nucleus,which can better prevent relapse.In the future,the degree of activation of the caudate nucleus under drug cues may serve as an indicator for evaluating the effectiveness of methadone treatment.

Objective To study the plasma metabolomics of mice poisoned by different dosage of the combination of diazepam and ethanol,and to reveal the toxicological mechanisms of combined poisoning of diazepam and ethanol.Methods Female Kunming mice were randomly divided into blank group,single and combined poisoning group(n=6),Based on the LD50 of diazepam co-administered with graded ethanol doses,mice in the single-drug and combined groups received oral gavage at 1/2,1,and 2 × LD50.Retro-orbital blood samples(～500 μL)were collected within 24 hours post-administration and analyzed by UPLC-QE-MS technology.Principal component analysis and orthogonal partial least squares discriminant analysis were used to identify differential metabolites and associated metabolic pathways.Results A total of 387 differential metabolites were identified in the combined poisoning group of diazepam and ethanol implicating the key pathways including tryptophan metabolism,phenylalanine metabolism,arginine and proline metabolism,Glycerophospholipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis.Conclusion Combined diazepam and ethanol poisoning exerts significant systemic effects by disrupting neurotransmitters conduction,exacerbating oxidative stress response and dysregulating energy metabolism.

OBJECTIVE: To observe the efficacy and safety of moxibustion in treating patients with central obesity of phlegm-dampness constitution. METHODS: A total of 66 patients with central obesity of phlegm-dampness constitution were randomly assigned to a moxibustion group (n=33, 3 cases dropped out) and a sham moxibustion group (n=33, 4 cases dropped out). The moxibustion group received mild moxibustion combined with lifestyle intervention; the moxibustion was applied at Shenque (CV8) and bilateral Zusanli (ST36), 30 min per session, maintaining a local skin temperature of (43±1) ℃. The sham moxibustion group received simulated moxibustion combined with lifestyle intervention; the simulated moxibustion was applied at the same acupoints, with the same session length, but with a maintained skin temperature of (37±1) ℃. Both groups were treated once every other day, three times per week for 8 consecutive weeks. Obesity-related physical indicators (waist circumference, hip circumference, body weight, body fat percentage, body mass index [BMI]), constitution evaluation indicators (phlegm-dampness constitution conversion score, symptom score), the impact of weight on quality of life-lite (IWQOL-Lite), the hospital anxiety and depression scale (HADS), and the incidence of adverse events were measured before and after treatment, and after 4 weeks of follow-up. RESULTS: Compared with before treatment, both groups showed significant reductions in waist circumference, hip circumference, body weight, body fat percentage, BMI, phlegm-dampness constitution conversion score and symptom score, IWQOL-Lite, and both anxiety and depression subscale scores of HADS after treatment and at follow-up (P<0.001). These improvements were significantly greater in the moxibustion group than those in the sham moxibustion group (P<0.001, P<0.01, P<0.05). One patient in the moxibustion group experienced a mild burn that resolved with routine care; the incidence of adverse reactions was 3.0% (1/33) in the moxibustion group and 0% (0/33) in the sham moxibustion group, with no statistically significant difference (P>0.05). CONCLUSION On the basis of lifestyle intervention, moxibustion effectively improves obesity-related physical indicators, enhances quality of life, alleviates anxiety and depression, and improves the phlegm-dampness constitution in patients with central obesity. These benefits persist for at least 4 weeks after treatment.
Humans ; Moxibustion ; Male ; Female ; Middle Aged ; Adult ; Obesity, Abdominal/psychology* ; Acupuncture Points ; Treatment Outcome ; Aged ; Quality of Life ; Young Adult ; Body Mass Index

Functional constipation is a highly prevalent gastrointestinal disorder that can significantly impact the quality of life of affected children.The intestinal flora，known as normal microorganisms in the human gut，interacts with various systems to maintain intestinal stability.Recent evidence has increasingly highlighted a relationship between dysbiosis and functional constipation.Intestinal flora may not only regulate intestinal motility by influencing the enteric nervous system，but also secrete a variety of metabolites that stimulate intestinal chemoreceptors to enhance motility，although the exact mechanisms remain unclear.The treatment of functional constipation through the regulation of intestinal flora has been widely studied，primarily focusing on probiotics and fecal microbiota transplantation.This article reviews the research progress related to intestinal flora and functional constipation.

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Objective To study the plasma metabolomics of mice poisoned by different dosage of the combination of diazepam and ethanol,and to reveal the toxicological mechanisms of combined poisoning of diazepam and ethanol.Methods Female Kunming mice were randomly divided into blank group,single and combined poisoning group(n=6),Based on the LD50 of diazepam co-administered with graded ethanol doses,mice in the single-drug and combined groups received oral gavage at 1/2,1,and 2 × LD50.Retro-orbital blood samples(～500 μL)were collected within 24 hours post-administration and analyzed by UPLC-QE-MS technology.Principal component analysis and orthogonal partial least squares discriminant analysis were used to identify differential metabolites and associated metabolic pathways.Results A total of 387 differential metabolites were identified in the combined poisoning group of diazepam and ethanol implicating the key pathways including tryptophan metabolism,phenylalanine metabolism,arginine and proline metabolism,Glycerophospholipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis.Conclusion Combined diazepam and ethanol poisoning exerts significant systemic effects by disrupting neurotransmitters conduction,exacerbating oxidative stress response and dysregulating energy metabolism.

Objective To explore the impact of the duration of protracted abstinence(PA)treatment on the level of impulsivity among heroin addicts and to analyze the mediating effect of impulsive cognition between abstinence duration and impulsive behavior.Methods Thirty-six heroin addicts undergoing PA treatment(PA group)and forty matched healthy controls(HC)(HC group)were recruited.Demographic information was collected via questionnaires,and impulsive cognition assessments were conducted.Participants completed the GO/NOGO task to collect data on impulsive behavior.Intergroup differences were compared using the independent sample t-test,and multiple linear regression analysis and mediating effect analysis were performed on the PA group.Results The PA group exhibited significantly higher scores on motor impulsivity,non-planning impulsivity,and total impulsivity compared to the HC group.Additionally,the PA group demonstrated significantly increased NOGO error rate during the GO/NOGO task compared to the HC group(P＜0.001).Multiple linear regression analysis revealed that abstinence duration had a significant negative impact on total impulsivity and non-planning impulsivity(P＜0.001).Mediating effect analysis found a partial mediation effect of total impulsivity between abstinence duration and NOGO error rate(P＜0.001).Conclusion The PA treatment has a significant impact on the level of impulsivity among heroin addicts.Impulsive cognition play a partial mediating role between abstinence duration and impulsive behavior.These results provide an important theoretical and practical basis for optimizing PA treatment programs.

Objective To explore the effects of different daily doses of methadone maintenance treatment(MMT)on subjective craving and brain function under drug cues among heroin addicts,providing an objective basis for the formulation of methadone medi-cation plan in clinical practice.Methods Twenty-nine heroin addicts were included and grouped according to the daily dose of metha-done 40 mg,≤40 mg/d group 15 participants(group A),>40 mg/d group 14 participants(group B).The functional magnetic reso-nance imaging(fMRI)data of brain response induced by drug cues were collected using a 3.0T MR scanner,and the craving data induced by drug cues in the subjects were collected.Brain activity and behavioral data processing were analyzed using SPM8 and SPSS 20.0 software.Results There were no statistically significant differences in craving scores between the two groups of subjects exposed to drug cues(t=-0.69,P>0.05).The brain regions with differences in brain response included the left caudate nucleus(group A showed signifi-cantly enhanced response),and the strength of its response was negatively correlated with the daily dose of methadone(r=-0.465,P=0.025).Conclusion Higher doses of MMT may help control the value cognitive processing of drug cues in heroin addicts through the caudate nucleus,which can better prevent relapse.In the future,the degree of activation of the caudate nucleus under drug cues may serve as an indicator for evaluating the effectiveness of methadone treatment.

Objective To explore the impact of the duration of protracted abstinence(PA)treatment on the level of impulsivity among heroin addicts and to analyze the mediating effect of impulsive cognition between abstinence duration and impulsive behavior.Methods Thirty-six heroin addicts undergoing PA treatment(PA group)and forty matched healthy controls(HC)(HC group)were recruited.Demographic information was collected via questionnaires,and impulsive cognition assessments were conducted.Participants completed the GO/NOGO task to collect data on impulsive behavior.Intergroup differences were compared using the independent sample t-test,and multiple linear regression analysis and mediating effect analysis were performed on the PA group.Results The PA group exhibited significantly higher scores on motor impulsivity,non-planning impulsivity,and total impulsivity compared to the HC group.Additionally,the PA group demonstrated significantly increased NOGO error rate during the GO/NOGO task compared to the HC group(P＜0.001).Multiple linear regression analysis revealed that abstinence duration had a significant negative impact on total impulsivity and non-planning impulsivity(P＜0.001).Mediating effect analysis found a partial mediation effect of total impulsivity between abstinence duration and NOGO error rate(P＜0.001).Conclusion The PA treatment has a significant impact on the level of impulsivity among heroin addicts.Impulsive cognition play a partial mediating role between abstinence duration and impulsive behavior.These results provide an important theoretical and practical basis for optimizing PA treatment programs.