ObjectiveTo investigate the effect of solute carrier family 7 member 5 (SLC7A5) inhibitor JPH203 on renal fibrosis induced by unilateral ureteral obstruction in mice. MethodsSixteen SPF male C57BL/6 mice were randomly divided into the control group and the experimental group, with 8 mice in each group. The mouse model of renal fibrosis was established by unilateral ureteral obstruction. From the third day after surgery, the mice in the control group were intraperitoneally injected with phosphate-buffered saline (PBS) for 11 consecutive days, and the injection dose was 200 μL/d. Mice in the experimental group received intraperitoneal injection of JPH203 (50 mg/kg) every day for 11 days. On day 14, the mice were euthanized, then the kidney tissues were obtained. Hematoxylin and eosin (HE) staining was used to assess renal tissue damage, Masson staining was used to evaluate collagen fiber deposition in the extracellular matrix, and immunohistochemistry was used to detect the levels of fibroblast activation markers α-smooth muscle actin (α-SMA) and collagen type Ⅰ (COL-Ⅰ) in kidney tissues. Western blotting was further performed to measure the expression levels of SLC7A5 and transforming growth factor-β1 (TGF-β1), as well as the phosphorylation levels of mammalian target of rapamycin complex 1 (mTORC1) signaling pathway-related molecules. Real-time quantitative PCR was used to verify changes in the mRNA levels of SLC7A5, α-SMA, and COL-Ⅰ in kidney tissues. ResultsCompared with the control group, the experimental group showed reduced destruction of renal tissue structure and a significantly lower pathological injury score (P<0.05). Additionally, collagen deposition in the extracellular matrix was decreased, and the percentage of collagen fiber area was significantly reduced (P<0.001) in the experimental group. The levels of fibroblast activation markers α-SMA and COL-Ⅰ were significantly lower in the experimental group (both P<0.001). The expression levels of SLC7A5 and TGF-β1 were also significantly decreased (P<0.001), and the phosphorylation levels of mTORC1 signaling pathway-related proteins 4E-BP1 and mTORC1 were significantly reduced (P<0.001). Real-time quantitative PCR confirmed that the mRNA levels of SLC7A5, α- SMA, and COL-Ⅰ in kidney tissues were significantly lower in the experimental group (P<0.001). ConclusionJPH203 may inhibit the progression of renal fibrosis in mice by suppressing SLC7A5 expression, regulating the mTORC1 signaling pathway, and altering fibroblast activation status.

The circadian clock plays a critical role in regulating various physiological processes, including gene expression, metabolic regulation, immune response, and the sleep-wake cycle in living organisms. Post-translational modifications (PTMs) are crucial regulatory mechanisms to maintain the precise oscillation of the circadian clock. By modulating the stability, activity, cell localization and protein-protein interactions of core clock proteins, PTMs enable these proteins to respond dynamically to environmental and intracellular changes, thereby sustaining the periodic oscillations of the circadian clock. Different types of PTMs exert their effects through distincting molecular mechanisms, collectively ensuring the proper function of the circadian system. This review systematically summarized several major types of PTMs, including phosphorylation, acetylation, ubiquitination, SUMOylation and oxidative modification, and overviewed their roles in regulating the core clock proteins and the associated pathways, with the goals of providing a theoretical foundation for the deeper understanding of clock mechanisms and the treatment of diseases associated with circadian disruption.
Protein Processing, Post-Translational/physiology* ; Circadian Rhythm/physiology* ; Humans ; Animals ; CLOCK Proteins/physiology* ; Circadian Clocks/physiology* ; Phosphorylation ; Acetylation ; Ubiquitination ; Sumoylation

This research investigated the contamination level,distribution of drug-resistant strains,and molecular epidemiologi-cal characteristics of Campylobacter,and further explored transmission pathways and prevention strategies.Cecum,chicken carcass,chicken product,and environmental samples,as well as swabs from workers'hands,were collected from a slaughterhouse in a large broiler group in the Jiaodong area between August 2023 and July 2024.Quantitative contamination assessment of Campylobacter in chicken carcasses and chicken products was performed.After microbial mass spectrometry identification,the representative strains of different links were selected for drug resistance testing and whole genome sequencing(WGS).On the basis of the sequencing results,the resistance genes,virulence genes,multilocus sequence typing(MLST),and phylogenetic characteristics of representative strains were analyzed.Homology comparisons were performed between isolates and strains from patients with diarrhea in the NCBI database.A total of 297 Campylobacter strains were isolated from 806 samples,and the overall detection rate was 36.85%.The detection rate of Campylobacter was highest in the evisceration process(47.33%),followed by the cutting process(35.64%).Overall,the Campylo-bacter detection rate first increased,then decreased,and subsequently increased.Drug sensitivity testing revealed that 90 isolates were resistant to nalidixic acid and ciprofloxacin,and 94.97%of isolates were resistant to tetracycline.WGS showed that both Campylo-bacter jejuni(C.jejuni)and Campylobacter coli(C.coli)carried many drug resistance and virulence genes.ST-14176 of C.jejuni was isolated for the first time herein.The predominant ST-8261 strain of C.jejuni and ST-860,ST-829,and ST-1586 strains of C.coli are known to cause human diarrhea.LOS expression genes associated with Guillain-Barré syndrome(GBS)were detected in both C.jejuni isolates from the slaughter chain and patients with GBS.Some strains exhibited close genetic relatedness to human-derived Campylo-bacter strains from the NCBI database.The detection rate of Campylobacter in the slaughterhouse first increased,then decreased,and subsequently increased,and the quantitative contamination level of each link was similar to the detection rate.Quantitative analysis of chicken carcasses/products revealed that the average bacterial load was highest in eviscerated carcasses(102.80 cfu/g),and the high-est amount of Campylobacter in chicken products reached 451.80 cfu/g.Abundant drug resistance genes and virulence genes were iden-tified,and the drug resistance genes were highly correlated with the drug resistance rate.Therefore,surveillance intensity and control measures for Campylobacter in slaughter processes should be strengthened.

OBJECTIVE: To explore the clinical significance of circulating tumor DNA (ctDNA) in response evaluation and relapse monitoring for patients with primary mediastinal large B-cell lymphoma (PMBCL). METHODS: The clinical characteristics, efficacy and survival of 38 PMBCL patients in our hospital from January 2010 to April 2020 were retrospectively analyzed. The ctDNA monitoring was conducted by targeted next-generation sequencing (NGS). RESULTS: Among the 38 patients, 26 cases were female, and 32 cases were diagnosed with Ann Arbor stage I-II. The 5-year overall survival (OS) rate and progression-free survival (PFS) rate were 74.7% and 61.7%, respectively. Males and those with high aaIPI scores (3 points) had a relatively poor prognosis. The NGS results of 23 patients showed that STAT6 (65.2%), SOCS1 (56.5%), and TNFAIP3 (56.5%) were the most common mutated genes. Patients with stable disease (SD)/progressive disease (PD) exhibited enrichment in cell cycle, FoxO, and TNF signaling pathways. A total of 29 patients underwent end-of-treatment PET/CT (EOT PET/CT), and 16 of them received ctDNA monitoring with 12 negative. Among 6 patients with EOT PET/CT positive (Deauville 4), 4 underwent ctDNA monitoring, and 3 of them were negative, being still in continuous remission without any subsequent anti-tumor therapy. CONCLUSION CtDNA may be combined with PET/CT to assess efficacy, monitor relapse, and guide treatment of PMBCL.
Humans ; Circulating Tumor DNA/blood* ; Female ; Mediastinal Neoplasms ; Male ; Retrospective Studies ; High-Throughput Nucleotide Sequencing ; Prognosis ; Lymphoma, Large B-Cell, Diffuse/genetics* ; Middle Aged ; Adult ; Aged ; Neoplasm Recurrence, Local ; Mutation

AIM:This study investigated the potential mechanism of acupuncture regulating adenosine A1 re-ceptor(ADORA1)in the mouse caudate putamen(CPu)to improve neuroplasticity and alleviate inflammatory pain.METHODS:Male C57BL/6 mice were randomly divided into five groups,namely,saline,complete Freund's adjuvant(CFA),acupuncture(MA),acupuncture+ADORA1 shRNA(MA+shRNA),and acupuncture+negative shRNA(MA+NCshRNA)groups.Twenty-one days before modeling,the mice in the MA+shRNA and MA+NCshRNA groups were in-jected with ADORA1 shRNA and control virus into the CPu.Modeling was performed 21 d later by injection of CFA into the right plantar skin of mice in the model and acupuncture groups to induce adjuvant-mediated arthritis.On day 2 after modeling,mice in the acupuncture groups received acupuncture at bilateral"Zusanli"points,30 min per session,once a day,for a total of 7 days.The paw thermal radiation pain threshold was used as an indicator of the effects on pain in the mice.Changes in the protein levels of ADORA1,synaptophysin(SYP),synapsin(SYN)I,SYN II,and brain-derived neurotrophic factor(BDNF)in the CPu were assessed by Western blot,and transmission electron microscopy was used to examine the dendritic structure and synaptic ultrastructure of neurons within the CPu.RESULTS:(1)Compared with the saline group,CFA modeling significantly reduced the thermal radiation pain threshold in mice(P<0.01),as well as the protein levels of ADORA1(P<0.01).Compared with the CFA group,the thermal pain threshold in the MA group in-creased between days 1 and 7 of acupuncture(P<0.05 or P<0.01),and ADORA1 protein levels increased(P<0.01).(2)Compared with the saline group,SYN I,SYN II,and BDNF protein levels were increased in the CFA group(P<0.05 or P<0.01),while relative to the CFA group,the levels of SYN I,SYN II,and BDNF were reduced in the acupuncture group(P<0.05 or P<0.01).No significant changes in SYP protein levels were observed in any of the groups.(3)Golgi staining and Sholl analysis showed that compared with the saline group,there were reductions in the total dendritic length and number of intersection points in the CFA group,while the dendritic spine density increased(P<0.05).Relative to the CFA group,the total dendritic length and the number of intersection points were increased in the MA group,while the dendritic spine density decreased(P<0.05).(4)Transmission electron microscopy revealed that compared with the Sa-line group,the synaptic clefts were narrower in the CFA group,while the density of synaptic vesicles in the presynaptic membrane was increased.Compared with the CFA group,synaptic clefts in the MA group were wider,and synaptic vesi-cle densities in the presynaptic membrane were reduced.(5)Twenty-one days after viral transfection,the thermal pain threshold in the MA and MA+NCshRNA groups increased from day 1 to day 7 of acupuncture relative to the CFA and MA+shRNA groups(P<0.05 or P<0.01),while the expression of ADORA1 was increased in both the MA and MA+NCshRNA groups(P<0.05 or P<0.01).(6)Compared with the CFA and MA+shRNA groups,the protein expression of SYN II and BDNF in the MA and MA+NCshRNA groups was reduced(P<0.05 or P<0.01).No significant changes in SYN I protein were observed in any of the groups.CONCLUSION:Acupuncture on Zusanli point can alleviate chronic pain in CFA-treated mice,potentially mediated by up-regulation of ADORA1 expression in the CPu brain region,thereby improving neuroplasticity.

Objective:To investigate the effect of tumor necrosis factor α (TNF-α) on the permeability of blood labyrinth barrier in C57BL/6J male mice and its possible mechanism.Methods:As for the design of animal experiment, twenty male C57BL/6J mice aged 6-8 weeks were randomly divided into control group and cisplatin group with 10 mice in each group by software method. The control group was intraperitoneally injected with normal saline every day, and the cisplatin group was intraperitoneally injected with 4 mg/kg cisplatin for 4 consecutive days. After administration, auditory brainstem response (ABR) was used to detect hearing changes in mice. HE staining was used to observe the morphological changes of mouse cochlea vasculature. The expression of TNF-α was detected by immunohistochemistry and ELISA. The permeability of the blood labyrinth barrier was observed by Evans blue staining. With respect to the design of cell experiment, primarily cultured cochlea pericytes (PC) and endothelial cells (EC) were divided into EC group, EC+TNF-α group, EC+PC group, EC+PC+TNF-α group, EC+PC+TNF-α+SB-3CT (MMP-9/MMP-2 secretion inhibitor) group, PC group, PC+TNF-α group, PC+TNF-α+LY294002 (PI3K/AKT pathway inhibitor) group, PC+LY294002 group. The protein expressions of ZO-1, VE-cadherin, MMP-9, MMP-2, PI3K, p-PI3K, AKT, and p-AKT were detected by Western blot. TEER and FITC-dextran penetration experiment were used to detect EC resistance and monolayer EC permeability, respectively. The data were statistically analyzed using GraphPad Prism 8 software.Results:In animal experiment, compared with control group, the ABR response threshold of mice in cisplatin group was increased ( P<0.01). The vaccine ular structure of the cochlea was disordered red, wrinkled and vacuole increased. The extravasation of vascular red fluorescent dye increased ( P<0.05), and also, levels of serum TNF-α and cochlear veins increased ( P<0.01). In cell experiment, by treatment of EC with different concentrations of cisplatin, 20 μmol cisplatin led to the highest expression of TNF-α ( P<0.01). The expression of TNF-α was the highest after 3 h intervention in EC ( P<0.05). Compared with those in EC+PC group, the resistance value of EC was decreased, the permeability of monolayers EC was increased, the expression of ZO 1 and VE cadherin proteins was decreased, and however, the resistance value was increased and the permeability of EC was decreased after the intervention of SB-3CT in EC+PC+TNF-α group. The expressions of ZO-1 and VE-cadherin proteins were increased ( P<0.05). The expression of MMP-9 increased after TNF-α intervention ( P<0.05), the expression of MMP-2 had no significant change, and the expression of p-PI3K/PI3K and p-AKT/AKT were increased ( P<0.05). The expression of MMP-9 decreased in PC+TNF-α+LY294002 group ( P<0.05). Conclusion:The hearing loss of C57BL/6J male mice induced by cisplatin may be related to the increased release of TNF-α from the blood labyrinth barrier EC in the cochlear stria vascularis, and the activation of PI3K/AKT signaling pathway by TNF-α in PC, which increases the secretion of MMP-9 from PC, ultimately leads to the increased permeability of the blood labyrinth barrier.

Chronic pain has emerged as a prevalent medical challenge in contemporary society.Patients suffering from chronic pain frequently develop comorbid psychological disorders,including anxiety,depression,post-traumatic stress disorder,and various psychiatric syndromes.These psychological complications not only affect patients' pain perception and responses,but may also constitute critical obstacles during pain management interventions.Acupuncture is a long-established clinical practice that has demonstrated remarkable efficacy in alleviating diverse pain types and has shown favorable therapeutic outcomes in ameliorating emotional disturbances such as anxiety and depression.The precise mechanisms underlying acupuncture-induced analgesia and anxiolytic effects,however,remain to be fully elucidated.In this context,it is essential to establish suitable and stable animal models to allow in-depth investigations into the pathogenesis of pain-related emotional disorders and the mechanistic foundations of acupuncture.This article presents a comprehensive review of recent literature regarding the selection of experimental animals,model-establishment method ologies,and behavioral-assessment paradigms pertaining to animal model platforms of chronic pain with comorbid anxiety.We also provide an in-depth discussion of research advancements regarding acupuncture intervention parameters,including needling techniques,acupoint selection,treatment duration,and efficacy evaluation within these animal models.This review proposes comprehensive and reference strategies for constructing preclinical animal models to investigate the mechanisms of acupuncture in managing chronic pain with comorbid anxiety,thus supporting scientific advancements in related research fields.

This research investigated the contamination level,distribution of drug-resistant strains,and molecular epidemiologi-cal characteristics of Campylobacter,and further explored transmission pathways and prevention strategies.Cecum,chicken carcass,chicken product,and environmental samples,as well as swabs from workers'hands,were collected from a slaughterhouse in a large broiler group in the Jiaodong area between August 2023 and July 2024.Quantitative contamination assessment of Campylobacter in chicken carcasses and chicken products was performed.After microbial mass spectrometry identification,the representative strains of different links were selected for drug resistance testing and whole genome sequencing(WGS).On the basis of the sequencing results,the resistance genes,virulence genes,multilocus sequence typing(MLST),and phylogenetic characteristics of representative strains were analyzed.Homology comparisons were performed between isolates and strains from patients with diarrhea in the NCBI database.A total of 297 Campylobacter strains were isolated from 806 samples,and the overall detection rate was 36.85%.The detection rate of Campylobacter was highest in the evisceration process(47.33%),followed by the cutting process(35.64%).Overall,the Campylo-bacter detection rate first increased,then decreased,and subsequently increased.Drug sensitivity testing revealed that 90 isolates were resistant to nalidixic acid and ciprofloxacin,and 94.97%of isolates were resistant to tetracycline.WGS showed that both Campylo-bacter jejuni(C.jejuni)and Campylobacter coli(C.coli)carried many drug resistance and virulence genes.ST-14176 of C.jejuni was isolated for the first time herein.The predominant ST-8261 strain of C.jejuni and ST-860,ST-829,and ST-1586 strains of C.coli are known to cause human diarrhea.LOS expression genes associated with Guillain-Barré syndrome(GBS)were detected in both C.jejuni isolates from the slaughter chain and patients with GBS.Some strains exhibited close genetic relatedness to human-derived Campylo-bacter strains from the NCBI database.The detection rate of Campylobacter in the slaughterhouse first increased,then decreased,and subsequently increased,and the quantitative contamination level of each link was similar to the detection rate.Quantitative analysis of chicken carcasses/products revealed that the average bacterial load was highest in eviscerated carcasses(102.80 cfu/g),and the high-est amount of Campylobacter in chicken products reached 451.80 cfu/g.Abundant drug resistance genes and virulence genes were iden-tified,and the drug resistance genes were highly correlated with the drug resistance rate.Therefore,surveillance intensity and control measures for Campylobacter in slaughter processes should be strengthened.

AIM:This study investigated the potential mechanism of acupuncture regulating adenosine A1 re-ceptor(ADORA1)in the mouse caudate putamen(CPu)to improve neuroplasticity and alleviate inflammatory pain.METHODS:Male C57BL/6 mice were randomly divided into five groups,namely,saline,complete Freund's adjuvant(CFA),acupuncture(MA),acupuncture+ADORA1 shRNA(MA+shRNA),and acupuncture+negative shRNA(MA+NCshRNA)groups.Twenty-one days before modeling,the mice in the MA+shRNA and MA+NCshRNA groups were in-jected with ADORA1 shRNA and control virus into the CPu.Modeling was performed 21 d later by injection of CFA into the right plantar skin of mice in the model and acupuncture groups to induce adjuvant-mediated arthritis.On day 2 after modeling,mice in the acupuncture groups received acupuncture at bilateral"Zusanli"points,30 min per session,once a day,for a total of 7 days.The paw thermal radiation pain threshold was used as an indicator of the effects on pain in the mice.Changes in the protein levels of ADORA1,synaptophysin(SYP),synapsin(SYN)I,SYN II,and brain-derived neurotrophic factor(BDNF)in the CPu were assessed by Western blot,and transmission electron microscopy was used to examine the dendritic structure and synaptic ultrastructure of neurons within the CPu.RESULTS:(1)Compared with the saline group,CFA modeling significantly reduced the thermal radiation pain threshold in mice(P<0.01),as well as the protein levels of ADORA1(P<0.01).Compared with the CFA group,the thermal pain threshold in the MA group in-creased between days 1 and 7 of acupuncture(P<0.05 or P<0.01),and ADORA1 protein levels increased(P<0.01).(2)Compared with the saline group,SYN I,SYN II,and BDNF protein levels were increased in the CFA group(P<0.05 or P<0.01),while relative to the CFA group,the levels of SYN I,SYN II,and BDNF were reduced in the acupuncture group(P<0.05 or P<0.01).No significant changes in SYP protein levels were observed in any of the groups.(3)Golgi staining and Sholl analysis showed that compared with the saline group,there were reductions in the total dendritic length and number of intersection points in the CFA group,while the dendritic spine density increased(P<0.05).Relative to the CFA group,the total dendritic length and the number of intersection points were increased in the MA group,while the dendritic spine density decreased(P<0.05).(4)Transmission electron microscopy revealed that compared with the Sa-line group,the synaptic clefts were narrower in the CFA group,while the density of synaptic vesicles in the presynaptic membrane was increased.Compared with the CFA group,synaptic clefts in the MA group were wider,and synaptic vesi-cle densities in the presynaptic membrane were reduced.(5)Twenty-one days after viral transfection,the thermal pain threshold in the MA and MA+NCshRNA groups increased from day 1 to day 7 of acupuncture relative to the CFA and MA+shRNA groups(P<0.05 or P<0.01),while the expression of ADORA1 was increased in both the MA and MA+NCshRNA groups(P<0.05 or P<0.01).(6)Compared with the CFA and MA+shRNA groups,the protein expression of SYN II and BDNF in the MA and MA+NCshRNA groups was reduced(P<0.05 or P<0.01).No significant changes in SYN I protein were observed in any of the groups.CONCLUSION:Acupuncture on Zusanli point can alleviate chronic pain in CFA-treated mice,potentially mediated by up-regulation of ADORA1 expression in the CPu brain region,thereby improving neuroplasticity.

Chronic pain has emerged as a prevalent medical challenge in contemporary society.Patients suffering from chronic pain frequently develop comorbid psychological disorders,including anxiety,depression,post-traumatic stress disorder,and various psychiatric syndromes.These psychological complications not only affect patients' pain perception and responses,but may also constitute critical obstacles during pain management interventions.Acupuncture is a long-established clinical practice that has demonstrated remarkable efficacy in alleviating diverse pain types and has shown favorable therapeutic outcomes in ameliorating emotional disturbances such as anxiety and depression.The precise mechanisms underlying acupuncture-induced analgesia and anxiolytic effects,however,remain to be fully elucidated.In this context,it is essential to establish suitable and stable animal models to allow in-depth investigations into the pathogenesis of pain-related emotional disorders and the mechanistic foundations of acupuncture.This article presents a comprehensive review of recent literature regarding the selection of experimental animals,model-establishment method ologies,and behavioral-assessment paradigms pertaining to animal model platforms of chronic pain with comorbid anxiety.We also provide an in-depth discussion of research advancements regarding acupuncture intervention parameters,including needling techniques,acupoint selection,treatment duration,and efficacy evaluation within these animal models.This review proposes comprehensive and reference strategies for constructing preclinical animal models to investigate the mechanisms of acupuncture in managing chronic pain with comorbid anxiety,thus supporting scientific advancements in related research fields.