Fatty liver, which is induced by abnormal lipid metabolism, is one of the most common causes of chronic liver disease globally and causes liver fibrosis. During this process, bone marrow-derived mesenchymal stromal cells (BMSCs) and hepatic stellate cells (HSCs) migrate toward the injured liver and participate in fibrogenesis by transdifferentiating into myofibroblasts. S100A8/A9 is a powerful inducer of cell migration and is involved in liver injury. But there are few reports about the effects of S100A8/A9 on BMSC/HSC migration. In the current study, we found that S100A8/A9 expression was increased during fatty liver injury/fibrogenesis. Moreover, S100A8/A9 expression had a positive correlation with fibrosis marker gene expressions in the injured liver. S100A8/A9 was mainly produced by neutrophils in the fibrotic liver. In vitro, neutrophil-secreted S100A8/A9 promoted BMSC/HSC migration via remodeling of microfilaments. Using specific siRNA and inhibitor, we proved that S100A8/A9-induced BMSC/HSC migration is dependent on TLR4/Rho GTPases signaling. Moreover, S100A8/A9 knock-down alleviated liver injury and fibrogenesis in vivo, while injection of S100A9 neutralizing antibody performed similar roles. We proved that S100A8/A9 was involved in liver injury and fibrogenesis via inducing BMSC/HSC migration. Our research reveals a new mechanism underlying BMSC/HSC migration in liver fibrosis and suggests S100A8/A9 as a potential therapeutic target of liver fibrosis. KEY MESSAGES: S100A8/A9 is secreted by neutrophils and increased in fatty liver injury. Neutrophil-secreted S100A8/A9 is a mediator of BMSC/HSC migration in vitro. S100A8/A9-induced BMSC/HSC migration is dependent on TLR4/Rho GTPases signaling. S100A8/A9 blockade alleviates liver injury and fibrogenesis in vivo.

BACKGROUND:To compare the efficacy of combined treatment of Er:YAG laser (ERL) and low-level laser therapy (LLLT) with single laser applications, and scaling and root planing (SRP) for non-surgical periodontal treatment. METHODS:In a randomized controlled trial, 25 non-smoking Stage II or Stage III periodontitis patients were recruited. The four intraoral quadrants were randomly assigned to four different treatments: (1) combined application with ERL plus SRP plus LLLT; (2) ERL plus SRP; (3) SRP plus LLLT; and (4) SRP. We assessed periodontal indexes, including probing depth (PD), clinical attachment level (CAL), bleeding index (BI), and plaque index (PLI), along with three cytokines (IL-1β, TNF-α, IL-10) from gingival crevicular fluid and red complex pathogens from subgingival dental plaque at baseline, 3 months, and 6 months. RESULTS:For initial moderate pockets (4 mm ≤ PD ≤ 6 mm), quadrants treated with ERL+SRP+LLLT, ERL+SRP, and SRP+LLLT exhibited greater PD improvement compared to the control (SRP) quadrants at the 3-month follow-up (1.25 ± 1.06, 1.23 ± 1.12, 1.00 ± 1.21 vs. 0.98 ± 1.21 mm) and the 6-month follow-up (1.35 ± 1.06, 1.23 ± 1.17, 1.35 ± 0.98 vs. 0.98 ± 1.23 mm) (p = 0.002). Quadrants treated with ERL+SRP+LLLT and SRP+LLLT showed more CAL gain means than the control quadrants at the 3-month follow-up (0.96 ± 1.42, 0.61 ± 1.39 vs. 0.55 ± 1.57 mm) and the 6-month follow-up (0.84 ± 1.54, 0.89 ± 1.49 vs. 0.48 ± 1.68 mm) (p = 0.008). For initial deep pockets (PD ≥ 7 mm), the ERL+SRP+LLLT quadrants had more PD improvement and CAL gain compared to the control quadrants at follow-up. There were no significant differences in BI, PLI, inflammatory cytokines, and periodontal pathogens among the four groups. CONCLUSION:The combined application of ERL and LLLT demonstrated potential efficacy in reducing PD, particularly for deep pockets. PLAIN LANGUAGE SUMMARY:To compare the therapy effect of combined use of Er:YAG laser (ERL) and low level laser therapy (LLLT) with single laser applications, and traditional periodontal treatment (SRP). A total of 25 non smoking patients with periodontitis were involved, and their mouths were divided into four sections, each receiving a different treatment: ERL+SRP+LLLT, ERL+SRP, SRP+LLLT, and SRP. Clinical indexes and laboratory indicators were assessed at baseline, 3 months, and 6 months. After six months, for initial moderate pockets, combined laser group and single laser group showed better improvements than traditional group in reducing the depth of periodontal pockets and increasing attachment levels. But for initial deep pockets, only combined laser group showed better improvement than traditional group. There were no significant differences in bleeding, plaque, inflammation, or harmful bacterial levels among the groups. These findings suggest that the integration of Er:YAG laser and low level laser therapy into standard periodontal treatment may enhance the treatment's benefits in reducing pocket depth, especially for severe conditions.

Atopic dermatitis (AD) is a chronic disease characterized by relapsed eczema and intractable itch, and is often triggered by house dust mites (HDM). PAR2 is a G-protein coupled receptor on keratinocytes and may be activated by HDM to affect AD processes. We first established a HDM-derived AD mouse model in wild-type (WT) and Par2-/-mice. Single cell RNA sequencing of the diseased skins found a stronger cellular communication between the ligand macrophage migration inhibitory factor (MIF) from keratinocytes and its receptors on antigen-presenting cells, suggesting the critical role of MIF in AD. HDM-WT mice showed severer skin lesions and pathological changes with stronger immunofluorescence MIF signals in skin sections than HDM-Par2-/- mice. Primary keratinocytes from WT mice stimulated with HDM or SLIGRL (PAR2 agonist) secreted more MIF in cultured medium and induced stronger immunofluorescence MIF signals than those from Par2-/- mice. The skin section of HDM-WT mice showed higher immunofluorescence signals of P115 (relating to MIF secretion) and KIF13B (possibly relating to intracellular trafficking of MIF) than that of HDM-Par2-/- mice. Acetylation of α-tubulin increased after stimulation by SLIGRL in WT keratinocytes but not in Par2-/- keratinocytes. HDM-WT mice treated with the MIF antagonist ISO-1 displayed improvement of AD-like presentations and lower expressions of IL-4, IL-13, TSLP and Arg1 (a biomarker of M2 macrophage) mRNAs. We conclude that MIF is an important cytokine and is significantly increased in the AD model. PAR2 affects AD changes by regulating the expression, intracellular trafficking, and secretion of MIF in epidermis.

Abstract Background The association between periodontitis and post-bronchodilator lung function is unclear. We aimed to determine the associations between symptoms of severe periodontitis (SSP) and post-bronchodilator lung function in the Chinese population. Methods A cross-sectional study (China Pulmonary Health study) was conducted from 2012 to 2015 in a large Chinese nationally representative sample of 49,202 participants aged 20–89 years. Data on demographic characteristics and periodontal symptoms of participants were collected by questionnaire. Participants who had at least one of the two severe symptoms (tooth mobility and natural tooth loss) in the past year were defined to have SSP, which was set as one variable for analyses. Post-bronchodilator lung function data including forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC) were collected by spirometry. Results The values of post-FEV1, post-FVC and post-FEV1/FVC of the participants with SSP were all significantly lower than the participants without SSP (all p < 0.001). SSP were significantly associated with post-FEV1/FVC < 0.7 (p < 0.001). In the multiple regression analyses, SSP were still negatively associated with post-FEV1(b = -0.04, 95%CI (-0.05 -0.03), p < 0.001), post-FEV1/FVC (b = -0.45, 95%CI (-0.63, -0.28), p < 0.001) and significantly associated with post-FEV1/FVC < 0.7 (OR = 1.08, 95%CI 1.01—1.16, p = 0.03) after full adjustment for potential confounders. Conclusions Our data suggest that SSP were negatively associated with post-bronchodilator lung function in the Chinese population. Longitudinal cohort studies are needed to confirm these associations in the future.

Class Ⅲ reservoirs in Daqing Oilfield are poorly developed, with characteristics of low permeability, small pore throat size and high clay content.At present, they are in field test stage of chemical flooding development. In order to understand influence of reservoir on adsorption loss of injected chemical agents and ensure development effect of chemical flooding, static adsorption experiment of weak alkali ASP system in condition of natural oil sandstone is carried out to compare adsorption loss of surfactant, alkali and polymer in Class Ⅲ and Class Ⅱreservoirs. Lab physical simulation experiment and numerical simulation technology are used to evaluate influence of different surfactant concentrations on economic effect of weak alkali ASP flooding technology, and the system concentration is optimized and applied in the field.The results show that adsorption loss of surfactant after 3 times of adsorption in Class Ⅲ reservoirs is 52.1%, which is 17 percentage points higher than that in Class Ⅱ reservoirs.The adsorption amount of alkali and polymer in Class Ⅲ is similar to that in Class Ⅱ reservoirs.Optimized weak alkali ASP flooding system for Class Ⅲ reservoirs has surfactant mass fraction of 0.35% and alkali mass fraction of 1.2% in main slug, and surfactant mass fraction of 0.20%～0.25% and alkali mass fraction of 1.0% in auxiliary slug.Field test application of the optimization result of weak alkali ASP flooding in NW block achieves obvious oil production increasing and water cut reducing effect. The research has guiding significance for promotion of chemical flooding for Class Ⅲ reservoirs in Daqing Oilfield.

Purpose: There is insufficient information about the prevalence and risk factors of gastroesophageal reflux disease (GERD) in the Chinese adult population. We aimed to assess the prevalence and identify the risk factors of GERD in China. Methods: We collected data from a nationally representative sample (50,991 subjects) of Chinese adults from a large nation-wide cross-sectional survey. GERD was diagnosed by a standardized Chinese-language GERD questionnaire with a score of > 8. The demographic characteristics, comorbidities and periodontal factors of all participants were collected. Results: Fifty-thousands-one-hundred-eighty-three participants were finally included in this study. The overall prevalence of GERD was 5.6% (95% CI, 5.4-5.8%) among the general Chinese population aged 20 years or older. Women, smokers, and people with older age, BMI > 25.0 kg/m2, urban residence, lower education level or comorbidities were more prevalent with GERD (p < 0.001). Symptoms of severe periodontitis (OR = 1.40, 95% CI 1.28-1.52, p < 0.001) and lower frequency of tooth brushing (OR = 2.01, 95% CI 1.76-2.29, p < 0.001) were significantly associated with risk of GERD. Conclusion: Symptom-based GERD is highly prevalent in the Chinese population. Overweight and smoking are major preventable risk factors for GERD. Periodontal factors are novel potential risk factors for GERD and should be given more attention in GERD prevention.

Introduction: Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by relapsed eczema and serious pruritus. High-mobility group box 1 protein (HMGB1) is a nuclear-binding protein and serves as an alarmin to promote inflammatory responses. Methods: In this study, we established an AD mouse model by topical use of MC903 on ears and then used a specific HMGB1-binding peptide cIY8 and a HMGB1 inhibitor of glycyrrhizin to investigate HMGB1 on fibroblast activation in the pathogenesis of AD-like symptoms. Results: Topical use of cIY8 and oral use of glycyrrhizin significantly improved the MC903-induced AD-like symptoms and pathological changes of the ears and scratching behavior in an AD mouse model; cIY8 treatment inhibited the higher mRNAs of IL-1α, IL-4, IL-5, IL-13, and IL-31 in the ears. In human fibroblasts, HMGB1 caused nuclear translocation of NF-kB, and the nuclear translocation could be inhibited by pre-treatment of HMGB1 with cIY8, suggesting that NF-κB signaling pathway participates in the HMGB1-induced inflammation of AD in fibroblasts and that cIY8 effectively impedes the function of HMGB1. Glycyrrhizin inhibited the Ca2+ signaling induced by ionomycin in mouse primary fibroblasts. The fibroblast-related proteins of α-SMA, Hsp47, and vimentin and the pruritus-related proteins of IL-33 and periostin were increased in the ears of the AD mouse model, the ratio of EdU incorporation became higher in mouse fibroblasts treated with MC903, and the higher proliferation and inflammatory responses of the fibroblasts could be reversed by glycyrrhizin treatment. Conclusions: Fibroblast activation by HMGB1 is one of the critical processes in the development of inflammation and pruritus in the AD mouse model. The specific HMGB1-binding peptide cIY8 and the HMGB1 inhibitor glycyrrhizin inactivate skin fibroblasts to alleviate the inflammation and pruritus in the AD mouse model. Peptide cIY8 may be topically used to treat AD patients in the future.

Abstract Background The associations between periodontitis and post-bronchodilator lung function decline and chronic obstructive pulmonary disease (COPD) are not clear. We aimed to determine the associations between symptoms of severe periodontitis and pre- and post-bronchodilator lung function decline and COPD in the Chinese population. Methods A cross-sectional study was conducted in a large Chinese nationally representative sample of 49,202 participants aged 20–89 years. Data on demographic characteristics, symptoms of severe periodontitis included tooth mobility and natural tooth loss, frequency of tooth brushing and other information were collected. Pre- and post-bronchodilator lung function data were also collected. Results Symptoms of severe periodontitis were significantly associated with pre- and post- bronchodilator lung function variables decline in the multivariable adjusted multiple linear regression models (all p < 0.001), and were also associated with pre- and post-forced expiratory volume in 1 second (FEV1) / forced vital capacity (FVC) < 0.7 (odds ratios (OR) = 1.11, 95% confidence interval (CI) 1.05–1.19, p = 0.001 and OR = 1.08, 95%CI 1.01–1.16, p = 0.03) in the multivariable adjusted multiple logistic regression models. Never brush tooth is associated with pre- and post-FEV1/FVC decline and pre- and post-FEV1/FVC < 0.7 in the multivariable adjusted multiple regression models (all p < 0.01). Conclusions Our data suggest that symptoms of severe periodontitis included tooth mobility and natural tooth loss are both associated with pre- and post-bronchodilator lung function decline and COPD (post-FEV1/FVC < 0.7) in the Chinese population.

Objectives:To observe the protective effect of cathepsin B inhibition against apoptosis of acinar cells in the early management of pancreatic contusion and laceration in rats, which would provide evidence of a potential early therapeutic for pancreatic contusion and laceration. Methods:Twenty-four rats were assigned to 2 groups: 1) Model (n = 12) with an induced pancreatic injury of severity I-II and 2) CA074-V (n = 12): an induced pancreatic injury, severity I-II treated with the cathepsin B inhibitor CA074-me (0.01 mg/g) by intravenous administration through the caudal vein at 5 minutes post model establishment. The mice in these two groups were further randomly divided into 4 subgroups containing 3 rats each that were sacrificed for quantitation of apoptosis, immunohistochemistry of cathepsin B, and serum amylase and lipase measurements at different time points after model establishment (0, 3, 6, and 12 hours). Results:The percentage of apoptotic pancreatic acinar cells collected from the injured tissues were much lower in the CA074-V group than the Model group at 3 hours [9.25 +/- 3.94% vs. 64.76 +/- 26.47%, P < 0.10] and 6 hours [14.71 +/- 8.22% vs. 66.60 +/- 13.54%, P < 0.10] post model establishment. The percentage of cathepsin B-positive pancreatic acinar cells were much lower in the CA074-V group than in the Model group at 3 hours [31.07 +/- 12.02% vs. 69.16 +/- 5.71%, P < 0.10], 6 hours [24.84 +/- 0.93% vs. 47.06 +/- 0.91%, P < 0.10], and 12 hours [28.33 +/- 9.14% vs. 52.72 +/- 1.25%, P < 0.10] post model establishment. Conclusions:Early cathepsin B inhibition effectively blocked acinar cell apoptosis in an experimental rat model of pancreatic contusion and laceration.

Excessive neutrophils are recruited to damaged tissue and cause collateral injury under chronic inflammatory conditions. Sphingosine 1-phosphate (S1P) modulates kinds of physiological and pathological actions by inducing recruitment of various cell types through S1P receptors (S1PRs). This study aimed to detect the S1P/S1PRs-mediated effects on neutrophil recruitment during chronic liver inflammation. In present study, increased neutrophils originated from bone marrow (BM) were detected in liver tissue of BDL-treated mice. Hepatic sphingosine kinase 1 (SphK, S1P rate-limiting enzyme) or S1P levels positively correlated with neutrophil marker expression in liver of mice and patients. In vitro, expression of S1PR1, S1PR2 and S1PR3 were detected in both mouse BM neutrophils and differentiated human neutrophil-like (dHL60) cells. S1P powerfully boosted the migration and cytoskeletal remodeling of BM neutrophils through S1PR1 or S1PR2. Different from BM neutrophils, the migration and cytoskeletal remodeling of dHL60 cells were mediated by S1PR2 or S1PR3. S1PR2 blockade obviously attenuates neutrophil infiltration in bile duct ligation (BDL)-induced mouse liver injury. In conclusion, S1P/S1PRs system plays a pivotal role in neutrophil recruitment.

The application of erbium-doped yttrium aluminum garnet(Er∶YAG)laser, neodymium-doped yttrium-aluminum garnet(Nd∶YAG)laser and diode laser in the treatment of periodontitis has been increasing in recent years, but there is controversy about the efficacy of laser in non-operative treatment of periodontitis.This paper mainly describes the features and advantages of Er∶YAG laser, Nd∶YAG laser and diode laser in non-surgical periodontal treatment, and provides advice on the selection of laser types in dental treatment and insights on the treatment of periodontitis in elderly patients with common chronic diseases.

Objective To evaluate the clinical efficacy of haploidentical haematopoietic stem cell transplantation (haplo-HSCT) for the treatment of malignant infantile osteopetrosis (MIOP) and intermediate osteopetrosis. Methods Children with MIOP and IOP who underwent haplo-HSCT in Beijing Children’s Hospital, Capital Medical University, from January 2010 to May 2018 were retrospectively analysed. Data relating to the clinical manifestations, engraftment, and prognosis of the children were extracted from medical records. Results Twenty-seven patients, including 18 males and 9 females, with an onset age of 12 (0.04–72) months were enrolled in this study. The median time from diagnosis to transplantation was 4 (1–23) months. All patients received haplo-HSCT with a myeloablative conditioning regimen (including fludarabine, busulfan, and cyclophosphamide). Graft versus host disease (GVHD) prophylaxis was based on anti-human T lymphocyte porcine immunoglobulin/anti-human thymus globulin, methotrexate, and mycophenolate mofetil. The median observation time was 55.2 (0.3–126.2) months. By the end of follow-up, twenty patients survived and seven patients died. The 5 year overall survival rate was 73.9%. Stage I-II acute GVHD was observed in 20 patients, stage III GVHD in 1 patient and no patients had stage IV disease. Chronic GVHD was observed in 11 patients (40.7%) and was controlled by anti-GVHD therapy. Conclusions Haplo-HSCT was an effective treatment for MIOP and IOP, with a high survival rate and significantly improved clinical symptoms. For patients with a vision impairment before HSCT, the improvement was slow after transplantation. The incidence of GVHD was high but mild and was effectively controlled by appropriate treatment. These data indicated that haplo-HSCT was a feasible treatment for MIOP and IOP.

NLRP3 inflammasome-driven inflammation represents a key trigger for hepatic fibrogenesis during cholestatic liver injury. However, whether sphingosine 1-phosphate (S1P) plays a role in NLRP3 inflammasome priming and activation remains unknown. Here, we found that the expression of NLRP3 in macrophages and NLRP3 inflammasome activation were significantly elevated in the liver injured by bile duct ligation (BDL). In vitro, S1P promoted the NLRP3 inflammasome priming and activation via S1P receptor 2 (S1PR2) in bone marrow–derived monocyte/macrophages (BMMs). Focusing on BMMs, the gene silencing of Gα12 or Gα13 by specific siRNA suppressed NLRP3 inflammasome priming and pro-inflammatory cytokine (IL-1β and IL-18) secretion, whereas Gα(i/o) and Gαq were not involved in this process. The MAPK signaling pathways (P38, ERK, and JNK) mediated NLRP3 inflammasome priming and IL-1β and IL-18 secretion, whereas blockage of PI3K, ROCK, and Rho family had no such effect. Moreover, JTE-013 (S1PR2 inhibitor) treatment markedly reduced NLRP3 inflammasome priming and activation in BDL-injured liver. Collectively, S1P promotes NLRP3 inflammasome priming and pro-inflammatory cytokines (IL-1β and IL-18) secretion via the S1PR2/Gα(12/13)/MAPK pathway, which may represent an effective therapeutic strategy for liver disease. • Hepatic NLRP3 expression was significantly elevated in BMMs of BDL-injured mouse liver. • S1P promoted NLRP3 inflammasome priming and activation in BMMs, depending on the S1PR2/Gα(12/13)/MAPK pathway. • Blockade of S1PR2 by JTE-013 reduced NLRP3 inflammasome priming and activation inflammasome in vivo.

目的 研究不同血糖控制水平的糖尿病肾病(diabetic nephropathy,DN)患者的牙周状况及特点,探讨牙周炎症程度对糖尿病肾病患者血糖控制的作用.方法 纳入119例糖尿病肾病患者,分为两组,糖化血红蛋白(glycosylated hemoglobin,HbA1c)良好(HbA1c<7％)组27例和糖化血红蛋白不良(HbA1c≥7％)组92例.牙周检查内容包括探诊深度(probing depth,PD)、出血指数(blood index,BI)、临床附着丧失(clinical attachment loss,CAL)以及口内余留牙数,口腔健康问卷包括口腔健康行为等.血清检测指标包括HbA1c、空腹血糖、总胆固醇等.结果 与HbA1c<7％组的患者相比,HbA1c≥7％组患者的PD(3.99±1.05 vs 3.49±0.93,P=0.031)、BI(3.45±0.58 vs 3.10±0.67,P=0.001)、CAL(6.40±2.18 vs 5.37±1.99,P=0.003)、PD≥4 mm百分率(中位数:54.17％vs 38.15％,P=0.001),PD≥5mm百分率(中位数:24.13％vs 12.85％,P=0.028)增加,差异具有统计学意义.多因素Logistic回归分析结果表明,除空腹血糖增加与HbA1c控制不良高度相关外,牙周检查指标CAL增加与DN患者HbA1c控制不良亦高度相关(OR=1.36,95％CI:1.01-1.83,P=0.043).结论 糖尿病肾病患者的HbA1c控制情况与牙周炎症程度有关,CAL增加与糖尿病肾病患者HbA1c控制不良高度相关.

Nod-like receptor (NLR) family pyrin domain containing 3 (NLRP3) has been regarded as an important initiator or promoter in multiple inflammatory diseases. However, the relationship between cannabinoid receptor 1 (CB1) and macrophage NLRP3 inflammasome and the corresponding molecular mechanism in liver inflammation remain unclear. Mouse liver injury models were induced by carbon tetrachloride (CCl4) or methionine-choline-deficient and high fat (MCDHF) diet. Human liver tissues were obtained from patients with different chronic liver diseases. CB1 expression was increased in liver tissue and macrophages of CCl4- and MCDHF-treated mice, positively correlated with NLRP3. CB1 agonist ACEA (Arachiodonyl-2'-Chloroethylamide) promoted NLRP3 expression and NLRP3 inflammasome activation in macrophages. CB1 blockade with its antagonist AM281 reduced NLRP3 expression, inflammasome activation, and liver inflammation in CCl4- and MCDHF-treated mice. MicroRNA-30b-5p (miR-30b-5p), screened by the intersection of bioinformatics databases and downregulated miRNAs in injured liver, negatively correlated with NLRP3 in mouse and human liver. miR-30b-5p was involved in CB1-mediated activation of NLRP3 inflammasome in macrophages by directly targeting NLRP3. Importantly, administration of miR-30b-5p agomir targeted NLRP3 and attenuated liver inflammation in the injured liver. Altogether, CB1/miR-30b-5p axis modulates NLRP3 expression and NLPR3 inflammasome activation in macrophages during liver inflammation, which provides a potential target for liver disease.

NLR family pyrin domain containing 3 (NLRP3) inflammasome accompanies chronic liver injury and is a critical mediator of inflammation-driven liver fibrosis. Sphingosine 1-phosphate (S1P)/S1P Receptor (S1PR) signaling participates in liver fibrogenesis by affecting bone marrow (BM)-derived monocytes/macrophage (BMM) activation. However, the relationship between S1P/S1PR signaling and NLRP3 inflammasome in BMMs remains unclear. Here, we found significantly elevated gene expression of NLRP3 inflammasome components (NLRP3, pro-interleukin-1β, and pro-interleukin-18) and the activation of NLRP3 inflammasome significantly elevated during murine chronic liver injury induced by a bile duct ligation operation, a methionine-choline–deficient and high-fat diet, or carbon tetrachloride intraperitoneal injection. Moreover, the increased expression of sphingosine kinase 1 (SphK1), the rate-limiting synthetic enzyme of S1P, was positively correlated with NLRP3 inflammasome components in both patients and mouse model livers. Flow cytometry analysis and immunofluorescence staining showed BMMs contributed to the significant proportion of NLRP3+ cells in murine inflammatory livers, but not Kupffer cells, dendritic cells, endothelial cells, T cells, and hepatocytes. Focusing on macrophages, S1P promoted NLRP3 inflammasome priming and activation in a dose-dependent manner. Blockade of S1PR2 by JTE-013 (antagonist of S1PR2) or S1PR2-siRNA inhibited S1P-induced NLRP3 inflammasome priming and inflammatory cytokine (interleukin-1β and interleukin-18) secretion, whereas blockade of S1PR1 or S1PR3 had no such effect. in vivo, a β1,3-d-glucan-encapsulated siRNA particle (GeRP) delivery system is capable of silencing genes in macrophages specifically. Treatment with S1PR2 siRNA-GeRPs markedly reduced NLRP3 inflammasome priming and activation and attenuated liver inflammation and fibrosis. Together, the conclusions indicated that targeting macrophage S1PR2 retarded liver inflammation and fibrogenesis via downregulating NLRP3 inflammasome, which may represent an effective therapeutic strategy for chronic liver injury.

Neutrophils play an essential role in the control of inflammatory diseases. However, whether cannabinoid receptors (CBs) play a role in neutrophil chemotaxis and NETosis in sterile liver inflammation remains unknown. The expression of marker genes on neutrophils was characterized by FACS, immunofluorescence, qRT-PCR, and Western blot. The amount of neutrophils was significantly elevated from 7 days and reached the peak at 2 weeks in carbon tetrachloride (CCl4)-treated mouse liver. The mRNA expression of neutrophil marker Ly6G had positive correlation with CB1 and CB2 expression in injured liver. In vitro CBs were abundantly expressed in isolated neutrophils and CB1 agonist ACEA promoted the chemotaxis and cytoskeletal remodeling, which can be suppressed by CB1 antagonist AM281. Moreover, ACEA induced NETosis, myeloperoxidase release from lysosome and ROS burst, indicating neutrophil activation, via Gαi/o. Conversely, CB2 agonist JWH133 had no effect on neutrophil function. ROS and p38 MAPK signaling pathways were involved in CB1-mediated neutrophil function, and ROS was upstream of p38 MAPK. CB1 blockade in vivo significantly attenuated neutrophil infiltration and liver inflammation in CCl4-treated mice. Taken together, CB1 mediates neutrophil chemotaxis and NETosis via Gαi/o/ROS/p38 MAPK signaling pathway in liver inflammation, which represents an effective therapeutic strategy for liver diseases.

Inappropriate neutrophil infiltration and subsequent neutrophil extracellular trap (NET) formation have been confirmed to be involved in chronic inflammatory conditions. Fatty liver disease is an increasingly severe health problem worldwide and currently considered the most common cause of chronic liver disease. Sphingosine 1-phosphate (S1P), a product of membrane sphingolipid metabolism, regulates vital physiological and pathological actions by inducing infiltration and activation of various cell types through S1P receptors (S1PRs). Here, we seek to determine the S1PR-mediated effects on neutrophil activation during chronic liver inflammation. In this study, NETs are detected in the early stage of methionine-choline-deficient and a high-fat (MCDHF) diet-induced liver injury. NET depletion by deoxyribonuclease I intraperitoneal injection significantly protects liver from MCDHF-induced liver injury in vivo. Meanwhile, we show that levels of myeloperoxidase-DNA complex (NET marker) in the serum present positive correlation with sphingosine kinase1 (S1P rate-limiting enzyme) messenger RNA expression or S1P levels in the injured liver of MCDHF-fed mice. In vitro, S1PR2 participates in the redirection of neutrophil apoptosis to NETosis via Gαi/o, extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and reactive oxygen species signaling pathways. Moreover, S1PR2 knockdown in MCDHF-fed mice by S1PR2-siRNA intravenous injection significantly inhibits NET formation in damaged liver tissue and then alleviates hepatic inflammation and fibrosis. Conclusion: In the early stage of fatty liver disease, S1PR2-mediated neutrophil activation plays an important role in the evolvement of liver injury.

Purpose: To evaluate clinical periodontal status and microbiologic pathogens in patients with chronic obstructive pulmonary disease (COPD) and periodontitis.Patients and Methods: We conducted a case-control study of 60 periodontitis patients with COPD (case group) and 60 periodontitis patients with normal pulmonary function (control group).Their periodontal status and respiratory function were clinically examined.Real-time polymerase chain reaction assays were used to measure five dental pathogens and four respiratory pathogens in subgingival dental plaque.Spearman's rank correlation coefficients (r 2 ) were calculated to assess correlations of pathogens.Principal component analysis (PCA) was employed to assess the similarity of bacterial diversity between the two groups.Logistic regression was performed to examine the associations of periodontal variables and pathogens with COPD risk.Results: COPD patients had fewer remaining teeth, higher plaque index (PLI), and more severe site percentages of clinical attachment level (CAL) than the controls.Although COPD patients tended to have relatively higher ranked means of Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, and Haemophilus influenza than control participants, the differences were not significant.Some periodontal pathogens and respiratory pathogens were positively correlated with each other (r 2 =0.29 to 0.47, all P < 0.05).The PCA graph showed that the distributions of pathogens were more dispersed but less discriminated in the COPD group than those in the control group.PLI (P = 0.045) and CAL ≥ 5mm site percentages (P = 0.01) were significantly associated with an increased risk of COPD, while pathogens were not associated with COPD. Conclusion:Our results from this study do not indicate periodontal pathogens as potential predictors of COPD risk, despite significantly poor periodontal status associated with COPD.