ObjectiveTo investigate the mechanism by which adrenoceptor alpha 2A (Adra2a) regulates lipopolysaccharide (LPS)-induced inflammation in primary hepatocytes from lipopolysaccharide-binding protein (LBP) knockout mice (Lbp^-/-). MethodsPrimary hepatocytes from C57BL/6J and Lbp^-/- mice were isolated using a two-step perfusion method. An in vitro inflammatory model was established by LPS stimulation, and an in vivo inflammatory mouse model was established by intraperitoneal injection of LPS. The in vitro experiments were grouped as follows: Control group, LPS group, BRL+LPS group, OE-NC+LPS group, and OE-Adra2a+LPS group. The Control group served as the blank control. The LPS group involved stimulating primary hepatocytes with LPS. The BRL+LPS group involved pretreating primary hepatocytes with BRL-44408 maleate followed by LPS stimulation. The OE-NC+LPS group involved transfecting primary hepatocytes with an empty vector followed by LPS stimulation. The OE-Adra2a+LPS group involved transfecting primary hepatocytes with a lentivirus overexpressing Adra2a, followed by LPS stimulation. The in vivo experimental groups were divided into Control', LPS', BRL+LPS', OE-NC+LPS', and OE-Adra2a+LPS' groups. The Control' group served as the blank control. The LPS' group received intraperitoneal injection of LPS. The BRL+LPS' group received intraperitoneal injection of BRL-44408 maleate for pretreatment, followed by LPS injection. The OE-NC+LPS' group received intraperitoneal injection of empty vector for pretreatment, followed by LPS injection. The OE-Adra2a+LPS' group received intraperitoneal injection of a lentivirus overexpressing Adra2a for pretreatment, followed by LPS injection. Cell viability after Adra2a inhibition and overexpression was assessed via the Cell Counting Kit-8 (CCK-8) assay. RT-qPCR measured changes in gene expression levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) after Adra2a inhibition and overexpression. Western blotting was performed to detect Adra2a protein expression and phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase, and c-Jun N-terminal kinase (JNK) following LPS stimulation. ResultsIn vitro experiments revealed that LPS stimulation significantly decreased Adra2a protein expression in primary hepatocytes from C57BL/6J mice compared to the Control group (P<0.05), whereas it increased in primary hepatocytes from Lbp^-/- mice (P<0.001). Compared to the LPS group, the BRL+LPS group exhibited significantly increased cell viability (P<0.01), reduced TNF-α, IL-6, and IL-1β gene transcription levels (P<0.01, P<0.001, P<0.001), and decreased phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.01, P<0.001, P<0.001). Compared with the OE-NC+LPS group, the OE-Adra2a+LPS group showed significantly decreased cell viability (P<0.001), increased gene transcription levels of TNF-α, IL-6, and IL-1β genes (P<0.001, P<0.01, P<0.001), and elevated phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.001). In vivo experiments showed that, compared with the LPS' group, the BRL+LPS' group exhibited significantly reduced phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.01). In the OE-Adra2a+LPS' group, the phosphorylation levels of ERK1/2, p38, and JNK were significantly elevated compared to the OE-NC+LPS' group (P<0.01, P<0.001, P<0.01). ConclusionLPS stimulation can cause a significant increase in Adra2a protein expression in primary hepatocytes of Lbp^-/- mice. Adra2a protein can regulate the level of LPS-induced inflammation in primary hepatocytes of Lbp^-/- mice through the MAPK signaling pathway.

ObjectiveThis paper aims to investigate the mechanism by which Erchentang improves body weight in obese mice by regulating the AMP‑activated protein kinase （AMPK）/peroxisome proliferator‑activated receptor γ coactivator‑1α （PGC‑1α） signaling pathway and inducing browning of inguinal white adipose tissue （iWAT）. MethodsObese mouse models were established by feeding a high‑fat diet. After successful modeling， mice were randomly divided into a model group and low‑， medium‑， and high‑dose Erchentang groups （7.5， 15， 30 g·kg^-1）， with six mice in each group. Another six normal mice were set as the normal group. Mice in the treatment groups were administered with corresponding doses of the drug by gavage， while those in the normal and model groups were administered with an equal volume of pure water by gavage for four consecutive weeks. Obesity was evaluated by body weight and Lee's index. The levels of low‑density lipoprotein cholesterol （LDL‑C） and high‑density lipoprotein cholesterol （HDL‑C） in serum were detected by biochemical assays. The leptin content in serum was measured by enzyme‑linked immunosorbent assay （ELISA）. Hematoxylin and eosin （HE） staining was used to observe the pathological morphology of the liver and iWAT. Immunofluorescence staining was applied to detect the protein expression levels of glucose transporter 4 （GLUT4） in the liver and iWAT. Molecular docking was performed to simulate the binding affinity between the key components of Erchentang （nobiletin， diosmetin， naringenin） and the key pathway proteins AMPK and PGC‑1α. Western blot was used to detect the protein expression levels of uncoupling protein‑1 （UCP‑1）， AMPK， phosphorylated AMP-activated protein kinase （p‑AMPK）， and PGC‑1α in iWAT. ResultsCompared with those in the normal group， the mice in the model group showed significantly increased body weight and Lee's index， elevated levels of HDL‑C， LDL‑C， and leptin in serum， enlarged adipocytes in iWAT， down‑regulated protein expression levels of GLUT4 in iWAT and liver， and decreased protein expression levels of UCP‑1 and PGC‑1α in iWAT（P<0.05， P<0.01）， the expression level of p-AMPK / AMPK protein was up-regulated， but the difference was not statistically significant. Compared with those in the model group， the mice in the Erchentang groups with different doses exhibited significantly reduced body weight and Lee's index， decreased levels of HDL‑C， LDL‑C， and leptin in serum， smaller adipocytes in iWAT， up‑regulated GLUT4 protein expression levels in iWAT and liver， and increased protein expression levels of UCP‑1， p‑AMPK/AMPK， and PGC‑1α in iWAT （P<0.05， P<0.01）. Molecular docking results show that nobiletin， diosmetin， and naringenin have strong binding energies with both AMPK and PGC‑1α. ConclusionErchentang may improve body weight in obese mice by regulating the AMPK/PGC‑1α signaling pathway and inducing iWAT browning.

To address the problem of data silos in dental specialties caused by equipment heterogeneity, this study developed an Intelligent Internet of Things (IoT)-enabled dental chair platform (hereinafter referred to as the intelligent platform) based on the concept of medical-engineering integration. The platform adopts a three-tier chair-domain interconnection architecture: the bottom tier integrates multi-source sensors and standardized interfaces for automated data acquisition and linkage with hospital information systems; the middle tier provides clinic-level management and remote teaching collaboration; and the top tier employs a blockchain-based secure cloud database for resource allocation and data management. Clinical validation at The Affiliated Stomatological Hospital of Nanjing Medical University demonstrated that, compared with a control group from the same period in 2023, the trial group achieved a 38.0% increase in average daily patient visits (80.6±6.8 vs. 58.4±5.2, t=15.16, P<0.001), a 24.6% reduction in average treatment time [(36.1±6.3) min vs. (47.9±8.5) min, t=7.72, P<0.001], a 39.2% reduction in waiting time [23.3 (16.5, 30.1) min vs. 38.3 (28.3, 48.3) min, U=32.00, P<0.001], a 30.4% reduction in equipment idle rate [8.7% (5.1%, 12.3%) vs. 12.5% (7.4%, 17.6%), U=251.00, P=0.003], and an increase in patient satisfaction from 88.2% (1 519/1 723) to 94.3% (2 186/2 318) ( t=7.26, P<0.001). User research confirmed that the functions most favored by clinicians and patients were "dental chair parameter updating and clinical data integration" [74.7% (80/107)] and "chairside display of diagnostic images" [76.8% (119/155)], respectively. Looking forward, the intelligent platform has the potential to integrate artificial intelligence-assisted diagnosis and 5G-enabled multicenter collaboration to further expand its clinical applications and accelerate the digital transformation of dental healthcare.

Objective To investigate the role of cholinergic receptor muscarinic 3(Chrm3)in regulating lipopolysaccharide(LPS)-induced inflammation in peritoneal macrophages in lipopolysaccharide binding protein(LBP)-knockout(Lbp-/-)mice.Methods Peritoneal macrophages were isolated from wild-type and Lbp-/-mice to establish an LPS-induced inflammation model.Chrm3 expression in Lbp-/-mouse peritoneal macrophages was inhibited by 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide(4-damp)and small interfering(siRNA)and Chrm3 overexpression was achieved by lentivirus transfection.For 4-damp inhibition,cells were divided into control,LPS,and inhibitor groups,and for siRNA transfection,cells were divided into control,LPS,si-normal control group,and si-Chrm3 groups.For overexpression,cells were divided into control,LPS,negative control,and overexpression groups.Changes in Chrm3 in response to LPS stimulation were verified by Western blot.The effects of 4-damp,si-Chrm3,and lentivirus on cell inflammation and survival were confirmed by Cell Counting Kit-8,quantitative polymerase chain reaction,and Western blot assays.Results Chrm3 protein expression was significantly elevated in Lbp-/-peritoneal macrophages post-LPS stimulation(P＜0.001),whereas there was no notable change in wild-type cells.The cell survival rate was significantly increased in the 4-damp and si-Chrm3 groups(P＜0.05,P＜0.01),and cell survival was significantly reduced in the overexpression group(P＜0.01).Furthermore,4-damp and si-Chrm3 significantly reduced expression levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6(P＜0.01,P＜0.001),and phospho-extracellular signal-regulated kinase(p-ERK)(P＜0.01,P＜0.001),which are associated with cell damage and inflammation.In contrast,TNF-α,IL-1β,IL-6(P＜0.001),and p-ERK protein(P＜0.001)were significantly elevated in the overexpression group.Conclusions LPS stimulation upregulated the expression of Chrm3 and proinflammatory cytokines in Lbp-/-peritoneal macrophages.Specific downregulation of Chrm3 by 4-damp and si-Chrm3 significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-peritoneal macrophages,while upregulation of Chrm3 using overexpressing lentivirus significantly elevated the expression of related inflammatory factors.Chrm3 is implicated in the regulation of the LPS-induced inflammation response in peritoneal macrophages in Lbp-/-mice.

In the mammalian central nervous system (CNS), astrocytes are the ubiquitous glial cells that have complex morphological and molecular characteristics. These fascinating cells play essential neurosupportive and homeostatic roles in the healthy CNS and undergo morphological, molecular, and functional changes to adopt so-called 'reactive' states in response to CNS injury or disease. In recent years, interest in astrocyte research has increased dramatically and some new biological features and roles of astrocytes in physiological and pathological conditions have been discovered thanks to technological advances. Here, we will review and discuss the well-established and emerging astroglial biology and functions, with emphasis on their potential as therapeutic targets for CNS injury, including traumatic and ischemic injury. This review article will highlight the importance of astrocytes in the neuropathological process and repair of CNS injury.
Astrocytes/drug effects* ; Humans ; Animals ; Central Nervous System/pathology* ; Central Nervous System Diseases/physiopathology*

Objective To investigate the role of cholinergic receptor muscarinic 3(Chrm3)in regulating lipopolysaccharide(LPS)-induced inflammation in peritoneal macrophages in lipopolysaccharide binding protein(LBP)-knockout(Lbp-/-)mice.Methods Peritoneal macrophages were isolated from wild-type and Lbp-/-mice to establish an LPS-induced inflammation model.Chrm3 expression in Lbp-/-mouse peritoneal macrophages was inhibited by 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide(4-damp)and small interfering(siRNA)and Chrm3 overexpression was achieved by lentivirus transfection.For 4-damp inhibition,cells were divided into control,LPS,and inhibitor groups,and for siRNA transfection,cells were divided into control,LPS,si-normal control group,and si-Chrm3 groups.For overexpression,cells were divided into control,LPS,negative control,and overexpression groups.Changes in Chrm3 in response to LPS stimulation were verified by Western blot.The effects of 4-damp,si-Chrm3,and lentivirus on cell inflammation and survival were confirmed by Cell Counting Kit-8,quantitative polymerase chain reaction,and Western blot assays.Results Chrm3 protein expression was significantly elevated in Lbp-/-peritoneal macrophages post-LPS stimulation(P＜0.001),whereas there was no notable change in wild-type cells.The cell survival rate was significantly increased in the 4-damp and si-Chrm3 groups(P＜0.05,P＜0.01),and cell survival was significantly reduced in the overexpression group(P＜0.01).Furthermore,4-damp and si-Chrm3 significantly reduced expression levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6(P＜0.01,P＜0.001),and phospho-extracellular signal-regulated kinase(p-ERK)(P＜0.01,P＜0.001),which are associated with cell damage and inflammation.In contrast,TNF-α,IL-1β,IL-6(P＜0.001),and p-ERK protein(P＜0.001)were significantly elevated in the overexpression group.Conclusions LPS stimulation upregulated the expression of Chrm3 and proinflammatory cytokines in Lbp-/-peritoneal macrophages.Specific downregulation of Chrm3 by 4-damp and si-Chrm3 significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-peritoneal macrophages,while upregulation of Chrm3 using overexpressing lentivirus significantly elevated the expression of related inflammatory factors.Chrm3 is implicated in the regulation of the LPS-induced inflammation response in peritoneal macrophages in Lbp-/-mice.

Objective:To explore the efficacy and factors affecting the treatment of gastric cancer liver metastasis (GCLM) with immune checkpoint inhibitors (ICI).Methods:This is a retrospective cohort study. Clinical and pathological data of 588 patients with GCLM treated at the Department of General Surgery, First Medical Center, People′s Liberation Army General Hospital, from January 2018 to December 2022 were retrospectively collected. There were 491 males and 97 females, aged ( M(IQR)) 60(14) years (range: 18 to 86 years). Patients were divided into an ICI treatment group ( n=142) and a non-ICI treatment group ( n=446) based on whether they received ICI therapy. Clinical and pathological data between the two groups were compared using the χ2 test or Mann-Whitney U test. Propensity score matching (PSM) was performed with cT stage, cN stage, surgical treatment, targeted therapy, and biomarkers as covariates, using a 1∶1 nearest neighbor matching method with a caliper value of 0.2. Univariate and multivariate analyses were conducted using Cox proportional hazards regression models, with relevant variables selected through forward stepwise regression. Survival curves were plotted using the Kaplan-Meier method, and group differences were compared using the Log-rank test. Subgroup analysis was conducted to identify potential beneficiary populations for ICI through forest plots. Results:After PSM, 114 patients were included in each group, and there were no statistically significant differences in the baseline data between the two groups (all P>0.05). The results of Cox multivariate analysis after PSM showed that cN2-3 stage ( HR=1.348, 95% CI: 1.091 to 1.665, P=0.006) and peritoneal metastasis ( HR=1.877, 95% CI:1.360 to 2.590, P<0.01) were independent risk factors for survival in GCLM patients; radical surgery ( HR=0.391, 95% CI: 0.305 to 0.501, P<0.01), immunotherapy ( HR=0.630, 95% CI: 0.503 to 0.788, P<0.01), and deficient DNA mismatch repair (dMMR) or combined positive score (CPS)≥5 ( HR=0.454, 95% CI: 0.320 to 0.644, P<0.01) were independent protective factors for survival in GCLM patients. After PSM, the overall survival was 12.4 (13.0) months in the non-immunotherapy group and 17.6 (17.8) months in the immunotherapy group (Log-rank test: P=0.029). Subgroup analysis showed that female patients, those with primary tumors located in the upper stomach, cN2-3 stage, one liver metastasis, synchronous liver metastasis, receiving targeted therapy, and those with dMMR or CPS≥5 were more likely to benefit from ICI therapy (all P<0.05). Conclusions:ICI prolongs overall survival in GCLM patients. Female patients, those with primary tumors located in the upper stomach, cN2-3 stage, one liver metastasis, synchronous liver metastasis, receiving targeted therapy, and those with dMMR or CPS≥5 are more likely to benefit from ICI therapy.

To address the problem of data silos in dental specialties caused by equipment heterogeneity, this study developed an Intelligent Internet of Things (IoT)-enabled dental chair platform (hereinafter referred to as the intelligent platform) based on the concept of medical-engineering integration. The platform adopts a three-tier chair-domain interconnection architecture: the bottom tier integrates multi-source sensors and standardized interfaces for automated data acquisition and linkage with hospital information systems; the middle tier provides clinic-level management and remote teaching collaboration; and the top tier employs a blockchain-based secure cloud database for resource allocation and data management. Clinical validation at The Affiliated Stomatological Hospital of Nanjing Medical University demonstrated that, compared with a control group from the same period in 2023, the trial group achieved a 38.0% increase in average daily patient visits (80.6±6.8 vs. 58.4±5.2, t=15.16, P<0.001), a 24.6% reduction in average treatment time [(36.1±6.3) min vs. (47.9±8.5) min, t=7.72, P<0.001], a 39.2% reduction in waiting time [23.3 (16.5, 30.1) min vs. 38.3 (28.3, 48.3) min, U=32.00, P<0.001], a 30.4% reduction in equipment idle rate [8.7% (5.1%, 12.3%) vs. 12.5% (7.4%, 17.6%), U=251.00, P=0.003], and an increase in patient satisfaction from 88.2% (1 519/1 723) to 94.3% (2 186/2 318) ( t=7.26, P<0.001). User research confirmed that the functions most favored by clinicians and patients were "dental chair parameter updating and clinical data integration" [74.7% (80/107)] and "chairside display of diagnostic images" [76.8% (119/155)], respectively. Looking forward, the intelligent platform has the potential to integrate artificial intelligence-assisted diagnosis and 5G-enabled multicenter collaboration to further expand its clinical applications and accelerate the digital transformation of dental healthcare.

Objective:To explore the efficacy and factors affecting the treatment of gastric cancer liver metastasis (GCLM) with immune checkpoint inhibitors (ICI).Methods:This is a retrospective cohort study. Clinical and pathological data of 588 patients with GCLM treated at the Department of General Surgery, First Medical Center, People′s Liberation Army General Hospital, from January 2018 to December 2022 were retrospectively collected. There were 491 males and 97 females, aged ( M(IQR)) 60(14) years (range: 18 to 86 years). Patients were divided into an ICI treatment group ( n=142) and a non-ICI treatment group ( n=446) based on whether they received ICI therapy. Clinical and pathological data between the two groups were compared using the χ2 test or Mann-Whitney U test. Propensity score matching (PSM) was performed with cT stage, cN stage, surgical treatment, targeted therapy, and biomarkers as covariates, using a 1∶1 nearest neighbor matching method with a caliper value of 0.2. Univariate and multivariate analyses were conducted using Cox proportional hazards regression models, with relevant variables selected through forward stepwise regression. Survival curves were plotted using the Kaplan-Meier method, and group differences were compared using the Log-rank test. Subgroup analysis was conducted to identify potential beneficiary populations for ICI through forest plots. Results:After PSM, 114 patients were included in each group, and there were no statistically significant differences in the baseline data between the two groups (all P>0.05). The results of Cox multivariate analysis after PSM showed that cN2-3 stage ( HR=1.348, 95% CI: 1.091 to 1.665, P=0.006) and peritoneal metastasis ( HR=1.877, 95% CI:1.360 to 2.590, P<0.01) were independent risk factors for survival in GCLM patients; radical surgery ( HR=0.391, 95% CI: 0.305 to 0.501, P<0.01), immunotherapy ( HR=0.630, 95% CI: 0.503 to 0.788, P<0.01), and deficient DNA mismatch repair (dMMR) or combined positive score (CPS)≥5 ( HR=0.454, 95% CI: 0.320 to 0.644, P<0.01) were independent protective factors for survival in GCLM patients. After PSM, the overall survival was 12.4 (13.0) months in the non-immunotherapy group and 17.6 (17.8) months in the immunotherapy group (Log-rank test: P=0.029). Subgroup analysis showed that female patients, those with primary tumors located in the upper stomach, cN2-3 stage, one liver metastasis, synchronous liver metastasis, receiving targeted therapy, and those with dMMR or CPS≥5 were more likely to benefit from ICI therapy (all P<0.05). Conclusions:ICI prolongs overall survival in GCLM patients. Female patients, those with primary tumors located in the upper stomach, cN2-3 stage, one liver metastasis, synchronous liver metastasis, receiving targeted therapy, and those with dMMR or CPS≥5 are more likely to benefit from ICI therapy.

Background Arsenic exposure is a common and important environmental and occupational hazardous factor in China, and arsenic-induced insulin resistance (IR) has attracted widespread attention as a negative health outcome to the population. Objective To explore part of the mechanism of hepatic IR induced by arsenic exposure based on the peroxisome proliferators-activated receptors γ (PPARγ)/ glucose transporter 4 (GLUT4) pathway, and to investigate potential effects of Ginkgo biloba extract (GBE) on hepatic IR induced by arsenic exposure and associated mechanism of action. Methods The target of drug action was predicted by network pharmacology and verified by in vivo and in vitro experiments. In vivo experiments: 48 SPF C57BL/6J male mice were divided into 4 groups, including control group, 50 mg·L⁻¹ NaAsO₂ model group (NaAsO₂), 50 mg·L⁻¹ NaAsO₂+10 mg·kg⁻¹ GBE intervene group (NaAsO₂+GBE), and 10 mg·kg⁻¹ GBE group (GBE), 12 mice in each group. The animals were given free access to purified water containing 50 mg·L⁻¹ NaAsO₂, or given intraperitoneal injection of normal saline containing 10 mg·kg⁻¹ GBE once per week. After 6 months of exposure, blood glucose detection, intraperitoneal glucose tolerance test (IPGTT), and insulin tolerance test (ITT) were performed. Serum and liver tissues were collected after the mice were neutralized, liver histopathological sections were obtained, serum insulin levels, liver tissue glycogen content, glucose content were detected by enzyme linked immunosorbent assay (ELISA), and the expression of PPARγ and GLUT4 proteins was detected by Western blot (WB). In vitro experiments: HepG2 cells were divided into 4 groups, including control group, 8 μmol·L⁻¹ NaAsO₂ group (NaAsO₂), 8 μmol·L⁻¹ NaAsO₂ + 200 mg·L⁻¹ GBE intervene group (NaAsO₂+GBE), and 200 mg·L⁻¹ GBE group (GBE). The levels of glycogen and glucose were detected by ELISA, and the expression of PPARγ and GLUT4 proteins was detected by WB. Results A strong binding effect between GBE and PPARγ was revealed by network pharmacology. In in vivo experiments, the NaAsO₂ group exhibited an elevated blood glucose compared to the control group, and the NaAsO₂+GBE group showed a decreased blood glucose compared to the NaAsO₂ group (P<0.01). The histopathological sections indicated severe liver structural damage in the arsenic exposure groups (NaAsO₂ group and NaAsO₂+GBE group), with varying staining intensity, partial liver cell necrosis, and diffuse red blood cell appearance. Both results of in vitro and in vivo experiments showed a decrease in glycogen synthesis and glucose uptake in the NaAsO₂ groups compared to the control groups, which was alleviated in the NaAsO₂+GBE group (P<0.01). The results of WB revealed inhibited PPARγ expression and reduced GLUT4 levels on the cell membrane, and all these changes were alleviated in the NaAsO₂+GBE group (P<0.01). Conclusion This study findings suggest that GBE antagonizes arsenic exposure-induced hepatic IR by regulating the PPARγ/GLUT4 pathway, indicating that GBE has a protective effect on arsenic exposure-induced hepatic IR, and PPARγ may be a potential therapeutic target for arsenic exposure-induced hepatic IR.