OBJECTIVES: To investigate the therapeutic mechanism of 2,6-dimethoxy-1,4-benzoquinone (DMQ) for alleviating dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. METHODS: Eighteen male C57BL/6J mice were equally randomized into control group, DSS group and DMQ treatment group. In DSS and DMQ groups, the mice were treated with DSS in drinking water to induce UC, and received intraperitoneal injections of sterile PBS or DMQ (20 mg/kg) during modeling. The changes in body weight, disease activity index (DAI), colon length, spleen weight, and colon histological scores of the mice were examined, and the percentages of Th17 and IFN-γ⁺ CD8⁺ T cells in the mesenteric lymph nodes and spleen were analyzed using flow cytometry. The expressions of tight junction proteins (Occludin and ZO-1), proteins associated with inflammasome activation (caspase-1 and p20), IL-1β and TNF-α in the colon tissues were detected using Western blotting or ELISA. In the cell experiment, mouse bone marrow-derived macrophages (BMDMs) primed with lipopolysaccharide (LPS) were treated with DMQ, followed by stmulation with nigericin to activate the classical NLRP3 inflammasome pathway. In cultured human peripheral blood mononuclear cells (PBMCs) treated with either LPS alone or LPS plus nigericin, the effects of DMQ on inflammasome activation, pyroptosis, and cytokine release were evaluated via Western blotting, ELISA, and flow cytometry. RESULTS: In DSS-treated mice, DMQ treatment significantly alleviated DSS-induced body weight loss, colon shortening, spleen enlargement, and colon inflammation. The DMQ-treated mice showed significantly reduced percentages of Th17 cells and IFN-γ⁺ CD8⁺ T cells in the mesenteric lymph nodes and spleen, with increased occludin and ZO-1 expressions and decreased caspase-1 expression in the colon tissue. DMQ obviously inhibited classical NLRP3 inflammasome activation in mouse BMDMs and both the classical and alternative pathways of NLRP3 activation in human PBMCs, causing also suppression of caspase-1-dependent pyroptosis. CONCLUSIONS DMQ ameliorates DSS-induced UC in mice by inhibiting NLRP3 inflammasome activation.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Mice, Inbred C57BL ; Colitis, Ulcerative/metabolism* ; Dextran Sulfate/adverse effects* ; Male ; Inflammasomes/metabolism* ; Mice ; Benzoquinones/therapeutic use* ; Th17 Cells ; Caspase 1/metabolism*

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Objective To investigate the value of mean platelet volume/lymphocyte ratio(MPVLR)in predicting the occurrence of deep vein thrombosis(DVT)after artificial hip replacement surgery.Methods A total of 104 patients who underwent artificial hip replacement surgery in Affiliated Hospital of North Sichuan Medical College from November 2022 to November 2023 were selected as the research objects.According to whether or not they had DVT,104 patients were divided into DVT group(36 cases)and non-DVT group(68 cases).The mean platelet volume(MPV)and lymphocyte count(LYC)were measured by automatic blood cell analyzer in all patients after admission,and MPVLR was calculated.Receiver operating characteristic curve was used to analyze the predictive value of MPV,LYC,and MPVLR for the occurrence of DVT after artificial hip replacement surgery.Multivariate Logistic regression analysis was used to explore the influencing factors of DVT after artificial hip replacement surgery.Results MPV and MPVLR in VTE group were higher than those in non-VTE group(P＜0.05),and LYC was lower than that in non-VTE group(P＜0.05).The area under the curve(AUC)of MPV and LYC for predicting DVT after artificial hip arthroplasty were 0.774(95％CI:0.729-0.824)and 0.851(95％CI:0.806-0.901),respectively.AUC of MPVLR for predicting DVT after artificial hip replacement surgery was 0.943(95％CI:0.898-0.993).The body mass index and the proportion of postoperative bed rest time≥3 days in the DVT group were higher than those in the non-DVT group(P＜0.05).Body mass index ≥22.73 kg/m2(OR=1.923,95％CI:1.237-2.989),postoperative bed rest time≥3 days(OR=2.380,95％CI:1.380-4.104),MPV≥9.65 fL(OR=2.563,95％CI:1.548-4.241),LYC≤1.70× 109/L(OR=2.208,95％CI:1.404-3.472),MPVLR≥5.95(OR=2.942,95％CI:1.816-4.764)were the risk factors for DVT after artificial hip replacement surgery(P＜0.05).Conclusion High MPVLR is one of the risk factors for DVT after artificial hip replacement surgery.It could be used as an im-portant indicator to predict the occurrence of DVT after artificial hip replacement surgery,and its predictive performance is excellent.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

【Objective】 To improve the understanding and diagnosis and treatment level of ALK negative anaplastic large cell lymphoma (ALK-ALCL) by sharing the diagnosis and treatment process of a patient with ALK-ALCL treated in Hangzhou Bay Hospital of Ningbo. 【Methods】 The clinical data and diagnosis and treatment process of the patient were retrospectively analyzed, and relevant literature was reviewed. 【Results】 The patient was a young male, with recurrent gross hematuria and right low back pain as the initial symptoms.Imaging examination indicated bladder tumor.After resection, the tumor was reduced and confirmed to be ALK-ALCL.After chemotherapy and autologous hematopoietic stem cell transplantation, the patient’s condition continued to improve.During the follow-up, no recurrence was observed. 【Conclusion】 Primary ALK-ALCL in the bladder is very rare and prone to misdiagnosis and missed diagnosis in clinical practice.The successful diagnosis and treatment experience of this patient can provide clinical reference.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

Objective:To summarize the clinical characteristics of ulcerative colitis (UC) with primary sclerosing cholangitis (PSC) .Methods:The clinical characteristics, diagnosis and treatment of 6 UC patients with PSC admitted to Sir Run Run Shaw Hospital affiliated to Zhejiang University School of Medicine from May 2017 to May 2022 were analyzed retrospectively. Meanwhile "ulcerative colitis", "primary sclerosing colangitis" were used as retrieval words to search associated literatures from CNKI, Wanfang Data, and PubMed databases as of August 2022. Relevant literatures were reviewed and summarized.Results:There were 6 UC patients complicated with PSC, included 4 males and 2 females. Four UC cases were diagnosed before PSC and other 2 cases were diagnosed simultaneously. Six cases were all extensive colonic type (E3), 2 patients were rectal sparing, and 4 patients had backwash ileitis. Five patients had elevated alkaline phosphatase (AKP) levels. All the patients received oral treatment with ursodeoxycholic acid (UDCA). Two patients with concurrent cholangiocarcinoma died within one year of follow-up. A total of 75 relevant literatures were retrieved.Conclusions:Patients with UC complicated with PSC are often diagnosed with UC before PSC, and PSC is often discovered due to abnormal liver function. The main clinical manifestations include jaundice, abdominal pain and diarrhea, and some may be accompanied by symptoms such as fever, vomiting, and fatigue. There are clinical characteristics such as elevated AKP, pancolitis, rectal sparing, and backwash ileitis. UC complicated with PSC is more likely to develop cholangiocarcinoma.

This article reports a rare Crohn′s disease (CD) case complicated with deep vein thrombosis and Guillain-Barre syndrome (GBS). Through the diagnosis and treatment of the multidisciplinary team, the condition of patient is relieved with no recurrence. Prevention of venous thromboembolism, rare extraintestinal manifestations and rare side adverse of anti-tumor necrosis factor-α monoclonal antibodies and their mechanisms in CD patients were discussed for reference.

Objective:To investigate whether memantine hydrochloride (MEM) could promote the bactericidal effect of neutrophils against methicillin-resistant Staphylococcus aureus (MRSA) and the possible mechanism. Methods:Neutrophils were co-incubated with different concentrations of MEM and MRSA for 4 h. Then the cell lysates were collected and cultured on plate for survival bacteria counting. After co-incubation, the neutrophils were collected to detect the production of reactive oxygen species (ROS) and the release of neutrophil extracellular traps (NETs). A mouse model of MRSA infection was established, and then the mice were treated with or without MEM. Blood, spleen and kidney samples were collected from the mice for bacterial colony counting and blood procalcitonin (PCT) detection. In the 48 h survival experiment, the mice were first infected with MRSA, and then treated with MEM or PBS. The survival rates of the mice were calculated and the survival curves were drawn.Results:The number of MRSA co-cultured with neutrophils decreased significantly in the presence of MEM, and within a certain concentration range, the survival number of MRSA decreased with the increase of MEM concentration. Moreover, MEM could significantly promote the production of ROS by neutrophils and the formation of NETs. In vivo experiment showed that the concentration of PCT in mouse blood samples was lower in the MRSA+ MEM group than in the MRSA+ PBS group. The animal experiment also revealed that MEM significantly decreased the bacteria loads in mouse blood and organs and increased the 48 h survival rate after MRSA infection.Conclusions:MEM could significantly promote the bactericidal effect of neutrophils against MRSA, which might be related to the enhanced generation of ROS by neutrophils and the formation of NETs.

Objective:To summarize the clinical characteristics of ulcerative colitis (UC) with primary sclerosing cholangitis (PSC) .Methods:The clinical characteristics, diagnosis and treatment of 6 UC patients with PSC admitted to Sir Run Run Shaw Hospital affiliated to Zhejiang University School of Medicine from May 2017 to May 2022 were analyzed retrospectively. Meanwhile "ulcerative colitis", "primary sclerosing colangitis" were used as retrieval words to search associated literatures from CNKI, Wanfang Data, and PubMed databases as of August 2022. Relevant literatures were reviewed and summarized.Results:There were 6 UC patients complicated with PSC, included 4 males and 2 females. Four UC cases were diagnosed before PSC and other 2 cases were diagnosed simultaneously. Six cases were all extensive colonic type (E3), 2 patients were rectal sparing, and 4 patients had backwash ileitis. Five patients had elevated alkaline phosphatase (AKP) levels. All the patients received oral treatment with ursodeoxycholic acid (UDCA). Two patients with concurrent cholangiocarcinoma died within one year of follow-up. A total of 75 relevant literatures were retrieved.Conclusions:Patients with UC complicated with PSC are often diagnosed with UC before PSC, and PSC is often discovered due to abnormal liver function. The main clinical manifestations include jaundice, abdominal pain and diarrhea, and some may be accompanied by symptoms such as fever, vomiting, and fatigue. There are clinical characteristics such as elevated AKP, pancolitis, rectal sparing, and backwash ileitis. UC complicated with PSC is more likely to develop cholangiocarcinoma.