OBJECTIVE To evaluate the efficacy and safety of Weiyan Tongluo Granules in treating chronic atrophic gastritis(CAG)and explore its potential mechanisms.METHODS From June 2020 to December 2022,100 CAG patients with spleen defi-ciency and blood stasis syndrome were enrolled and randomly divided into a trial group(n=50)and a control group(n=50)using a random number table.The trial group received Weiyan Tongluo Granules plus a folic acid placebo,while the control group received fo-lic acid tablets plus a traditional Chinese medicine granule placebo.The treatment course for both groups was 24 weeks,with 8 and 10 dropouts in the trial and control groups,respectively.Post-treatment comparisons included OLGA/OLGIM staging reversal rates,low-grade intraepithelial neoplasia regression rate,SSDPRO-CG(Patient-Reported Outcome Scale for Chronic Gastritis in Spleen-Stomach Diseases)scores,TCM syndrome scores,and safety indicators.Serum levels of PG I,PGⅡ,PGR,and G-17 were measured via ELISA before and after treatment.Gastric mucosal p-NF-κB and CDX2 protein expression levels were analyzed by Western blot,while mRNA levels of IL-1β,IL-6,VIL1,and MUC2 were quantified via qPCR.RESULTS After treatment,the trial group showed sig-nificantly higher OLGA and OLGIM stage reversal rates than the control group(P<0.05,P<0.01),though no significant difference was observed in low-grade intraepithelial neoplasia regression.Both groups exhibited significant improvements in physiological domain scores and total SSDPRO-CG scores(P<0.01),with the trial group outperforming the control group in physiological,independence,psychological domains,and total scores(P<0.05,P<0.01).TCM syndrome scores(total and sub-items:gastric distension,pain,poor appetite,bloating)decreased significantly in both groups(P<0.01),while the trial group showed greater reductions in loose stools and dull complexion(P<0.01).After-treatment,the trial group had significantly lower TCM syndrome scores than the control group(P<0.05,P<0.01).Serum PG I,PGⅡ,PGR,G-17,gastric mucosal p-NF-κB,CDX2,and IL-1β,IL-6,VIL1,MUC2 mRNA levels improved significantly in the trial group(P<0.05,P<0.01),while the control group improved only in PGⅡ(P<0.05,P<0.01).The trial group's improvements in these biomarkers surpassed the control group's(P<0.05,P<0.01).No treatment-related adverse events occurred in either group.CONCLUSION Weiyan Tongluo Granules ameliorate gastric mucosal pathology,clinical symptoms,psychological state,and quality of life in CAG patients without significant adverse effects.Its mechanism may involve sup-pressing the NF-κB pathway to reduce IL-1β and IL-6 expression,downregulating CDX2 to inhibit VIL1 and MUC2 transcription,thereby reversing the vicious cycle of inflammation-intestinal metaplasia.

OBJECTIVE To evaluate the efficacy and safety of Weiyan Tongluo Granules in treating chronic atrophic gastritis(CAG)and explore its potential mechanisms.METHODS From June 2020 to December 2022,100 CAG patients with spleen defi-ciency and blood stasis syndrome were enrolled and randomly divided into a trial group(n=50)and a control group(n=50)using a random number table.The trial group received Weiyan Tongluo Granules plus a folic acid placebo,while the control group received fo-lic acid tablets plus a traditional Chinese medicine granule placebo.The treatment course for both groups was 24 weeks,with 8 and 10 dropouts in the trial and control groups,respectively.Post-treatment comparisons included OLGA/OLGIM staging reversal rates,low-grade intraepithelial neoplasia regression rate,SSDPRO-CG(Patient-Reported Outcome Scale for Chronic Gastritis in Spleen-Stomach Diseases)scores,TCM syndrome scores,and safety indicators.Serum levels of PG I,PGⅡ,PGR,and G-17 were measured via ELISA before and after treatment.Gastric mucosal p-NF-κB and CDX2 protein expression levels were analyzed by Western blot,while mRNA levels of IL-1β,IL-6,VIL1,and MUC2 were quantified via qPCR.RESULTS After treatment,the trial group showed sig-nificantly higher OLGA and OLGIM stage reversal rates than the control group(P<0.05,P<0.01),though no significant difference was observed in low-grade intraepithelial neoplasia regression.Both groups exhibited significant improvements in physiological domain scores and total SSDPRO-CG scores(P<0.01),with the trial group outperforming the control group in physiological,independence,psychological domains,and total scores(P<0.05,P<0.01).TCM syndrome scores(total and sub-items:gastric distension,pain,poor appetite,bloating)decreased significantly in both groups(P<0.01),while the trial group showed greater reductions in loose stools and dull complexion(P<0.01).After-treatment,the trial group had significantly lower TCM syndrome scores than the control group(P<0.05,P<0.01).Serum PG I,PGⅡ,PGR,G-17,gastric mucosal p-NF-κB,CDX2,and IL-1β,IL-6,VIL1,MUC2 mRNA levels improved significantly in the trial group(P<0.05,P<0.01),while the control group improved only in PGⅡ(P<0.05,P<0.01).The trial group's improvements in these biomarkers surpassed the control group's(P<0.05,P<0.01).No treatment-related adverse events occurred in either group.CONCLUSION Weiyan Tongluo Granules ameliorate gastric mucosal pathology,clinical symptoms,psychological state,and quality of life in CAG patients without significant adverse effects.Its mechanism may involve sup-pressing the NF-κB pathway to reduce IL-1β and IL-6 expression,downregulating CDX2 to inhibit VIL1 and MUC2 transcription,thereby reversing the vicious cycle of inflammation-intestinal metaplasia.

A 58-year-old male patient with angioimmunoblastic T-cell lymphoma developed a rash and skin tightness on the face, limbs, and trunk together with joint stiffness and dysfunction after 6 months of treatment with the programmed cell death protein-1 inhibitor camrelizumab. Laboratory tests revealed progressive eosinophilia over 6 months, with the eosinophil count increasing from 0.07×10 9/L to 3.3×10 9/L. Magnetic resonance imaging showed thickened skin of both forearms, while T 2-weighted imaging showed markedly increased signal intensity within the myofascia. Skin biopsy of the right forearm showed thickened and fibrosed fascia and infiltration of inflammatory cells, including lymphocytes, plasma cells, and eosinophils. The patient was diagnosed with immune checkpoint inhibitor (ICI)-induced eosinophilic fasciitis (EF). After beginning treatment with methylprednisolone (40 mg daily), methotrexate (10 mg/week), and baricitinib (4 mg daily), his symptoms of skin tightness and joint dysfunction significantly improved within 1 month, and his peripheral blood eosinophil count decreased to 0.17×10 9/L. ICI-induced EF is a rare immune-related adverse reaction. To date, only 20 cases have been reported in published foreign literature, and their clinical characteristics are summarized here. The time from ICI treatment to EF was 12 (8,15) months, and the main clinical manifestations included skin involvement ( n=19), joint dysfunction ( n=11), myalgia/muscle weakness ( n=9), and peripheral eosinophilia ( n=16). After treatment, the clinical symptoms of EF improved in 17 patients, and eosinophil counts returned to normal after 3 (1,8) months. EF is a dysfunctional adverse response to ICI therapy. Tumor patients undergoing immunotherapy should be monitored for symptoms of EF. Early treatment is essential for preventing complications.

Objective:To explore the facilitating and inhibitory factors influencing the behavior of young patients to share medical electronic word-of-mouth (eWOM) on internet platforms, so as to provide insights for the improvement of healthcare quality.Methods:In May 2022, 271 undergraduate students from universities in Zhejiang province were selected by convenient sampling to survey their motivations to share eWOM with a self-designed questionnaire. Multiple regression analysis was used to examine the impact of different motivational factors on the sharing intention of young patients.Results:Only 16 respondents (5.9%) had previously published medical eWOM. Egoistic motivation, altruistic motivation, medical experience, and comment habits were significant factors that promoted patients to share eWOM, with egoistic motivation ( β=0.212, P<0.001) having the greatest impact and comment habit ( β=0.139, P=0.003) having the least impact. Distrust, low self-efficacy and involvement, perceived reluctance, and perceived uselessness were significant factors inhibiting patients from publishing eWOM. Of them, distrust ( β=-0.161, P<0.001) and perceived reluctance ( β=-0.161, P=0.001) had the greatest impact, and low self-efficacy and involvement had the least impact ( β=-0.134, P=0.003). Conclusions:To enhance the positive attitude of young patients towards sharing eWOM, it is important to focus on their personal benefits and provide high-quality healthcare experiences. Building trust among patients in the platform is crucial, and efforts should be made to reduce operational barriers. Additionally, educating and raising awareness among young patients regarding the significance and influence of healthcare reviews is important.

Objective : To explore the differences in the effects of psoriatic serum and a mixture of five proinflam⁃ matory cytokines on keratinocyte inflammation and proliferation . Methods : he human immortalized keratinocyte cell line (HaCaT cells) was used to cultivate with serum of healthy human serum , psoriasis patients and M5 factors , and the cell proliferation was monitored by CCK⁃8 . RT⁃qPCR was used to detect the mRNA expression levels of inflammatory factors interleukin 1β/8/21 /23 (IL⁃1β/8/21 /23) , proliferation markers keratin 6/16 (K6/K16) and nucleus related antigen 67 (Ki67) in HaCaT cells in each group . Results : Healthy human serum , psoriasis serum and M5 factors could effectively promote the growth of HaCaT cells , and psoriasis serum and M5 factors could promote cell growth more than healthy human serum . Similar to M5 factors , serum from patients with psoriasis significantly promoted the mRNA levels of inflammatory factors IL⁃1β , IL⁃8 , IL⁃23 and proliferation markers K6 and Ki67 in cells . Different from M5 factors , the serum of psoriasis patients had enhanced the mRNA level of inflammatory factor IL⁃21 , but weakened the promotion of proliferation marker K16 . Conclusion The serum micro environment of patients with psoriasis can promote the proliferation and inflammatory response of keratinocytes higher than that of healthy people , and is similar to M5 factors , which can be used to build a keratinocyte model of psoriasis .

Purpose: Accumulating evidence has suggested that toll-like receptor 4 (TLR4) is critically involved in the pathogenesis of asthma. The aim of this study was to investigate the role of TLR4 in toluene diisocyanate (TDI)-induced allergic airway inflammation. Methods: TLR4−/− and wild-type (WT) C57BL/10J mice were sensitized and challenged with TDI to generate a TDI-induced asthma model. B-cell lymphoma 2 (Bcl-2) inhibitors, ABT-199 (4 mg/kg) and ABT-737 (4 mg/kg), were intranasally given to TDI-exposed TLR4−/− mice after each challenge. Results: TDI exposure led to increased airway hyperresponsiveness (AHR), granulocyte flux, bronchial epithelial shedding and extensive submucosal collagen deposition, which were unexpectedly aggravated by TLR4 deficiency. Following TDI challenge, TLR4−/− mice exhibited down-regulated interleukin-17A and increased colony-stimulating factor 3 in bronchoalveolar lavage fluid (BALF), while WT mice did not. In addition, TLR4 deficiency robustly suppressed the expression of NOD-like receptor family pyrin domain containing 3 and NLR family CARD domain containing 4, decreased caspase-1 activity in TDI-exposed mice, but had no effect on the level of high mobility group box 1 in BALF. Flow cytometry revealed that TDI hampered both neutrophil and eosinophil apoptosis, of which neutrophil apoptosis was further inhibited in TDI-exposed TLR4−/− mice, with marked up-regulation of Bcl-2. Moreover, inhibition of Bcl-2 with either ABT-199 or ABT-737 significantly alleviated neutrophil recruitment by promoting apoptosis. Conclusions These data indicated that TLR4 deficiency promoted neutrophil infiltration by impairing its apoptosis via up-regulation of Bcl-2, thereby resulting in deteriorated AHR and airway inflammation, which suggests that TLR4 could be a negative regulator of TDI-induced neutrophilic inflammation.

Purpose: Accumulating evidence has suggested that toll-like receptor 4 (TLR4) is critically involved in the pathogenesis of asthma. The aim of this study was to investigate the role of TLR4 in toluene diisocyanate (TDI)-induced allergic airway inflammation. Methods: TLR4−/− and wild-type (WT) C57BL/10J mice were sensitized and challenged with TDI to generate a TDI-induced asthma model. B-cell lymphoma 2 (Bcl-2) inhibitors, ABT-199 (4 mg/kg) and ABT-737 (4 mg/kg), were intranasally given to TDI-exposed TLR4−/− mice after each challenge. Results: TDI exposure led to increased airway hyperresponsiveness (AHR), granulocyte flux, bronchial epithelial shedding and extensive submucosal collagen deposition, which were unexpectedly aggravated by TLR4 deficiency. Following TDI challenge, TLR4−/− mice exhibited down-regulated interleukin-17A and increased colony-stimulating factor 3 in bronchoalveolar lavage fluid (BALF), while WT mice did not. In addition, TLR4 deficiency robustly suppressed the expression of NOD-like receptor family pyrin domain containing 3 and NLR family CARD domain containing 4, decreased caspase-1 activity in TDI-exposed mice, but had no effect on the level of high mobility group box 1 in BALF. Flow cytometry revealed that TDI hampered both neutrophil and eosinophil apoptosis, of which neutrophil apoptosis was further inhibited in TDI-exposed TLR4−/− mice, with marked up-regulation of Bcl-2. Moreover, inhibition of Bcl-2 with either ABT-199 or ABT-737 significantly alleviated neutrophil recruitment by promoting apoptosis. Conclusions These data indicated that TLR4 deficiency promoted neutrophil infiltration by impairing its apoptosis via up-regulation of Bcl-2, thereby resulting in deteriorated AHR and airway inflammation, which suggests that TLR4 could be a negative regulator of TDI-induced neutrophilic inflammation.

Objective:To study the cellular pathology and molecular mechanisms of intestinal mucosal damage induced by IgG immune complex in mice.And to explore the pathogenic mechanism and molecular diagnosis evidence of ulcerative colitis induced by food intolerance in clinical practice.Methods: Six weeks old BALB/c female mice were used to build animal model.All the mice were divided into four groups:the control group(group A),the rabbit intestinal mucosal protein immunized group(group B),the DSS induced group(group C),the rabbit intestinal mucosal protein immunized combined with DSS induced group(group D).After successful establishment of animal model,serum and colon tissues were collected to be performed relevant tests.Results: IgG level in serum and colonic mucosa of group B mice increased.And inflammatory cell infiltration and a small amount of mast cell activation were observed in intestinal mucosa;group C mice showed the typical acute ulcerative colitis:a large number of inflammatory cells infiltration,inflammatory factor levels increased in mucosa and mucosa lamina propria,and mucosal epithelial cells′ tight junction weakened;group D mice manifested both high level of IgG in serum and colonic mucosa and also typical acute ulcerative colitis.Besides,significant mast cell activation was observed in the intestinal mucosa.Conclusion: We infer from the experimental results that IgG immune complexes can induce the damage of intestinal epithelium by mediating activation of mast cells.And during the process,the level of inflammatory cytokines increased in intestinal mucosa and the expression of tight junction protein in epithelial cell decreased.These factors contribute to the promotion of intestinal mucosa damage induced by immune complexes.

PURPOSE: Cell-in-cell structures are created by one living cell entering another homotypic or heterotypic living cell, which usually leads to the death of the internalized cell, specifically through caspase-dependent cell death (emperitosis) or lysosome-dependent cell death (entosis). Although entosis has attracted great attention, its occurrence is controversial, because one cell line used in its study (MCF-7) is deficient in caspase-3. METHODS: We investigated this issue using MCF-7 and A431 cell lines, which often display cell-in-cell invasion, and have different levels of caspase-3 expression. Cell-in-cell death morphology, microstructures, and signaling pathways were compared in the two cell lines. RESULTS: Our results confirmed that MCF-7 cells are caspase-3 deficient with a partial deletion in the CASP-3 gene. These cells underwent cell death that lacked typical apoptotic properties after staurosporine treatment, whereas caspase-3-sufficient A431 cells displayed typical apoptosis. The presence of caspase-3 was related neither to the lysosome-dependent nor to the caspase-dependent cell-in-cell death pathway. However, the existence of caspase-3 was associated with a switch from lysosome-dependent cell-in-cell death to the apoptotic cell-in-cell death pathway during entosis. Moreover, cellular hypoxia, mitochondrial swelling, release of cytochrome C, and autophagy were observed in internalized cells during entosis. CONCLUSION: The occurrence of caspase-independent entosis is not a cell-specific process. In addition, entosis actually represents a cellular self-repair system, functioning through autophagy, to degrade damaged mitochondria resulting from cellular hypoxia in cell-in-cell structures. However, sustained autophagy-associated signal activation, without reduction in cellular hypoxia, eventually leads to lysosome-dependent intracellular cell death.
Apoptosis ; Autophagy ; Caspase 3* ; Cell Death ; Cell Hypoxia ; Cell Line ; Cytochromes c ; Entosis ; MCF-7 Cells* ; Mitochondria ; Mitochondrial Swelling ; Staurosporine

Objective:To analyse the physicochemical properties and structure of major privet pollen allergen Lig v 1 using bioinformatics software and provide a reference for choosing suitable recombinant expression system for Lig v 1 and modifying the allergen Lig v 1 experimentally.Methods:The physicochemical properties were analysed by ProtParam,the signal peptide by SignalP 4.1 Server,the transmembrane helix by TMHMM Server v.2.0,the secondary structure by GOR4,MHCⅡepitopes by NetMHCⅡ2.2 Server,B-cell epitopes by ProteanTM 5.01 and the phylogenetic tree by MEGA 6.Results: Privet major pollen allergen Lig v 1 was stable in Escherichia coli and it doesn′t possess any signal peptide and transmembrane helix.Most secondary structures of Lig v 1 were random coils.Potential region of MHCⅡepitope of Lig v 1 was 30-44.Potential B-cell epitopes possess discontinuous and continuous a mino acid sequences.Lig v 1 and its counterparts from Fraxinus excelsior and Olea europaea were clustered into one group.Conclusion:Escherichia coli is the suitable expression system for recombinant Lig v 1.In silico prediction of the epitopes of Lig v 1 provides a reference for modifying the allergen Lig v 1 experimentally.