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.

Recent data suggest that vascular endothelial growth factor receptor inhibitor (VEGFRi) can enhance the anti-tumor activity of the anti-programmed cell death-1 (anti-PD-1) antibody in colorectal cancer (CRC) with microsatellite stability (MSS). However, the comparison between this combination and standard third-line VEGFRi treatment is not performed, and reliable biomarkers are still lacking. We retrospectively enrolled MSS CRC patients receiving anti-PD-1 antibody plus VEGFRi (combination group, n=54) or VEGFRi alone (VEGFRi group, n=32), and their efficacy and safety were evaluated. We additionally examined the immune characteristics of the MSS CRC tumor microenvironment (TME) through single-cell and spatial transcriptomic data, and an MSS CRC immune cell-related signature (MCICRS) that can be used to predict the clinical outcomes of MSS CRC patients receiving immunotherapy was developed and validated in our in-house cohort. Compared with VEGFRi alone, the combination of anti-PD-1 antibody and VEGFRi exhibited a prolonged survival benefit (median progression-free survival: 4.4 vs. 2.0 months, P=0.0024; median overall survival: 10.2 vs. 5.2 months, P=0.0038) and a similar adverse event incidence. Through single-cell and spatial transcriptomic analysis, we determined ten MSS CRC-enriched immune cell types and their spatial distribution, including naive CD4⁺ T, regulatory CD4⁺ T, CD4⁺ Th17, exhausted CD8⁺ T, cytotoxic CD8⁺ T, proliferated CD8⁺ T, natural killer (NK) cells, plasma, and classical and intermediate monocytes. Based on a systemic meta-analysis and ten machine learning algorithms, we obtained MCICRS, an independent risk factor for the prognosis of MSS CRC patients. Further analyses demonstrated that the low-MCICRS group presented a higher immune cell infiltration and immune-related pathway activation, and hence a significant relation with the superior efficacy of pan-cancer immunotherapy. More importantly, the predictive value of MCICRS in MSS CRC patients receiving immunotherapy was also validated with an in-house cohort. Anti-PD-1 antibody combined with VEGFRi presented an improved clinical benefit in MSS CRC with manageable toxicity. MCICRS could serve as a robust and promising tool to predict clinical outcomes for individual MSS CRC patients receiving immunotherapy.
Humans ; Colorectal Neoplasms/drug therapy* ; Male ; Female ; Immunotherapy ; Middle Aged ; Aged ; Tumor Microenvironment/immunology* ; Retrospective Studies ; Microsatellite Instability ; Transcriptome ; Single-Cell Analysis ; Programmed Cell Death 1 Receptor/immunology* ; Gene Expression Profiling ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors*

Objective:To summarized the clinical characteristics and surgical management of anterior cervical enterogenic in pediatric patients. Methods:Clinical data were retrospectively analyzed for 9 children with pathologically confirmed anterior cervical enterogenic cysts（including bronchogenic and esophagogenic subtypes） treated at the Children's Hospital of Shandong University（Jinan Children's Hospital） between January 1, 2020, and November 30, 2023. Results:Nine patients（6 males and 3 females） were involved in this study, aged 14 days to 10 years old. There were 4 cases on the left side, 4 on the right side, and 1 in the middle of the neck. All patients presented with neck masses. The patients were followed up from 3 months to 35 months after surgery and recovered well, with no recurrence or complications observed. Conclusion:①Anterior intestinal cysts in children are rare and easy to be misdiagnosed. ②Concurrent branchial cleft fistulas or associated anomalies may coexist, necessitating comprehensive evaluation. ③Preoperative diagnosis is not easy and mainly depends on pathological diagnosis. ④The treatment of anterior cervical enterogenic cysts in children is surgical resection of the lesion.
Humans ; Male ; Female ; Child ; Retrospective Studies ; Child, Preschool ; Infant ; Neck ; Cysts/surgery*

Kupffer cells (KCs), as residents and sentinels of the liver, are involved in the formation of hepatic fibrosis (HF). However, the biological functions of circular RNAs (circRNAs) in KCs to HF have not been determined. In this study, the expression levels of circRNAs, microRNAs, and messenger RNAs (mRNAs) in KCs from a mouse model of HF mice were investigated using microarray and circRNA-Seq analyses. circDcbld2 was identified as a candidate circRNA in HF, as evidenced by its up-regulation in KCs. Silver staining and mass spectrometry showed that Wtap and Igf2bp2 bind to cirDcbld2. The suppression of circDcbld2 expression decreased the KC inflammatory response and oxidative stress and inhibited hepatic stellate cell (HSCs) activation, attenuating mouse liver fibrogenesis. Mechanistically, Wtap mediated the N ⁶-methyladenosine (m6A) methylation of circDcbld2, and Igf2bp2 recognized m6A-modified circDcbld2 and increased its stability. circDcbld2 contributes to the occurrence of HF by binding miR-144-3p/Et-1 to regulate the inflammatory response and oxidative stress. These findings indicate that circDcbld2 functions via the m6A/circDcbld2/miR-144-3p/Et-1 axis and may act as a potential biomarker for HF treatment.

In the context of the development of transplant oncology, it is of great clinical significance to find a drug with both antitumor and immunosuppressive effects for liver transplantation patients with hepatocellular carcinoma (HCC). The antitumor effect of ginkgolic acid (GA) has been confirmed, and some studies suggest that GA may also have an immunosuppressive effect. The immunosuppressive effect of GA was evaluated by histopathology, T-cell subpopulation, and cytokine detection in rat liver transplantation and mouse cardiac transplantation models, and transcriptomic and metabolomic analysis was used to explore the underlying mechanism of the GA immunosuppressive effect. Metabolites, activation, and ferroptosis markers of CD8⁺ T cells were detected in vivo and in vitro. Based on rat liver transplantation and mouse cardiac transplantation models, the immunosuppressive effect of GA was first confirmed by histopathology, T-cell subpopulation, and cytokine detection. In the mouse cardiac transplantation model, transcriptomics combined with metabolomics demonstrated for the first time that GA inhibited lactate dehydrogenase A (LDHA) expression and pyruvate metabolism in CD8⁺ T cells. It was confirmed in vivo and in vitro that GA inhibited pyruvate metabolism of CD8⁺ T cells through LDHA, inhibiting their activation and inducing ferroptosis. Overexpression of LDHA partially reversed the effect of GA on the metabolism, activation, and ferroptosis of CD8⁺ T cells in vitro. GA mediates metabolic reprogramming through LDHA to inhibit the activation and induce ferroptosis of CD8⁺ T cells to exert an immunosuppressive effect, which lays an experimental foundation for the future clinical application of its immunosuppressive effect.

The molecular mechanisms underlying the health-promoting effects of exercise remain to be fully elucidated. As a bridge between genetics, exercise and phenotype, metabolites can be detected in high throughput through metabolomics, offering valuable insights into mechanism elucidation and disease prediction. Metabolic homeostasis is intricately regulated by various factors, including enzyme activity and transporters. Integration of multiple omics technologies such as genomics, transcriptomics, and proteomics enables the comprehensive elucidation of the metabolic network modulated by exercise interventions and facilitates the identification of key metabolic markers. This review summarizes the current research advancements, biological functions, discovery methods, and applications of exercise-induced multi omics metabolic markers, furnishing a theoretical foundation for understanding the mechanisms of exercise-induced health benefits and enabling precision interventions. Relevant literatures from 2000 to 2025 were systematically retrieved from databases including PubMed, CNKI and other databases with the keywords such as “multi-omics”, “metabolic biomarkers”, “exercise”, “health”. Subsequently, the identified literature was meticulously screened to meet the specified criteria and was subsequently incorporated into the study. (1) Exercise induces profound alterations in metabolite levels within the body, with particular emphasis on markers associated with sugar, lipid, and protein metabolism being extensively investigated. As an intensity marker, lactate is implicated in the regulation of fat browning (UCP-1), angiogenesis (VEGF), mitochondrial function (PGC-1α) and metabolic homeostasis (HIF-1α/CES2). Following resistance training, pyruvate levels increase, and an aberrant pyruvate to lactate ratio (approximately 10) may indicate mitochondrial dysfunction. Supplementation with pyruvate has been shown to reduce weight and lipid levels. Ketone bodies regulate metabolism by inhibiting lipolytic enzyme activity and promoting insulin secretion. Plasma ketone body concentrations rise after high-intensity exercise, with levels positively associated with central fatigue. Carnitine levels elevate post-endurance training, and supplementation with carnitine has been linked to increased lean body mass and enhanced cognitive function in older individuals. Serum alanine levels rise following resistance training and, as a precursor of carnosine, supplementation can elevate carnosine concentration by 80%, exerting antioxidant and neuroprotective effects. Creatine, a pivotal molecule in phosphogen energy supply, exhibits a 93% increase in plasma levels post-marathon, with its metabolism intricately related to AMPK activation. (2) Metabolites play a crucial role in disease prediction, particularly in the context of cardiovascular disease where 18 metabolites including glycoprotein acetyl and ketone bodies have been shown to enhance the performance of prediction models. Similarly, in diabetes research, acylcarnitine and other metabolites can improve prediction model efficacy. The combination of multiple metabolites has been found to substantially enhance predictive capabilities for various conditions such as cancer, aging, and other risks, surpassing the predictive power of traditional indicators. (3) Genomics investigations have unveiled the genetic underpinnings of exercise-related metabolites. VO₂max, a significant exercise phenotype with heritability estimates ranging from 0.59 to 0.66, exhibits a negative correlation with the susceptibility to diabetes and cardiovascular disease. SNPs associated with VO₂max, such as variants in the FSHR gene, are positively linked to serum creatinine levels. Reduced creatinine levels have been associated with an elevated risk of T2DM. These findings suggest that creatinine serves as a potential marker of exercise metabolism. (4) Transcriptomic studies have elucidated the molecular mechanisms by which exercise modulates metabolites. Acute exercise induces rapid alterations in the expression profiles of 9 132 transcripts. Exercise elicits upregulation of genes involved in the fructose/mannose metabolic pathway (such as SORD, PFKFB3), suggesting these metabolites may serve as pivotal mediators in the beneficial effects of exercise on Parkinson’s disease. Altitude training enhances the expression of the PHOSPHO1 gene, which encodes an enzyme facilitating choline synthesis. Choline deficiency has been linked to insulin resistance. Choline supplementation has been shown to augment the effects of resistance training, underscoring the significance of choline as a key marker in exercise-mediated metabolic health promotion. (5) Proteomic analyses have unveiled the key mechanisms through which exercise modulates metabolism. Endurance training induces significant alterations in myofibrillar expression, with 237 slow muscles and 172 fast muscles proteins showing differential regulation, of which 65% are associated with metabolism, including ACSL1 and ECHS1. Various training modalities elicit distinct phosphorylation modifications, exemplified by the negative correlation between LDHA3 phosphorylation and lactate levels. Endurance training upregulates SLC25A15 expression in adipose tissue, enhancing arginine synthesis. The post-exercise elevation of plasma GPLD1 levels mimics the neuroprotective effects of exercise on the brain. These findings present novel targets for investigating exercise-related metabolic markers. The application of multi omics technologies has expedited the identification and mechanistic analysis of both established and novel sports-related metabolic markers like lactate. Integrated multi omics strategies (e.g., genome-metabolome) enable the simultaneous examination of metabolic markers and their regulatory mechanisms, facilitating the discovery of exercise-related genetic markers and pivotal regulatory proteins. However, challenges persist, including inadequate data integration and a lack of standardization. Future endeavors should focus on developing dynamic monitoring tools, integrating state-of-the-art approaches such as single-cell/spatial omics, and leveraging AI algorithms for optimized analysis to construct precise predictive models for maximizing health benefits in exercise.

Based on the discussion of "eight weak areas" in The Inner Canon of Yellow Emperor （《黄帝内经》）， combined with the typical rash manifestations of atopic dermatitis， it is believed that atopic dermatitis is mostly deficiency-excess complex， and that pathogens intruding eight weak areas are the core of its pathogenesis. The external cause is exterior deficiencies， with heat， wind， dampness and other pathogenic qi attacking. The heart， lungs， kidneys out of balance， and excess pathogen are the internal cause， in which fire constraint and excessive heat are the basis of the disease， the wind invading leads to the progress of the changes， dampness obstructing channels and colla-terals make the condition persistent. Internal and external pathogens combination and retention result to the course of the disease lingering and difficult to cure. The internal treatment is to regulate zang-fu organs， and the formula could use self-prescribed modified Qingrun Tongluo Decoction （清润通络汤）， clearing heart and reducing fire in order to clear the heat and cool the blood， moistening lungs and generating metal to consolidate the exterior and dispel the wind， and nourishing kidneys and draining water to dispel the dampness and activate the collaterals. The external treatment applies maceration， fire acupuncture， wrapping to dredge the eight weak areas and regulate qi and blood in channel， so as to expel pathogens.

ObjectiveThis paper aims to systematically collect and organize ancient and modern clauses and studies containing Xieqingwan， excavate and analyze the key information of Xieqingwan， and provide a reference for facilitating the development of the classic formula Xieqingwan. MethodsThe composition， dosage， decocting methods， usage， and other key information of Xieqingwan in ancient traditional Chinese medicine books were collected and analyzed by means of literature research and metrological methods. The modern clinical application of Xieqingwan was summarized. ResultsA total of 42 pieces of effective data involving 32 ancient traditional Chinese medicine books were collected. Xieqingwan was first recorded in Xiaoer Yaozheng Zhijue. The drug origin of this formula is basically clear in the ancient traditional Chinese medicine books. The modern drug usage and decocting method were as follows： Angelicae Sinensis Radix， Gentianae Radix et Rhizoma， Chuanxiong Rhizoma， Gardenia seeds， Radix et Rhizoma Rhei， Notopterygii Rhizoma et Radix， and Saposhnikoviae Radix were grounded to fine powder， decocted with honey， and finally formed into pills with the size of a chicken head （1.5 g）. It was suggested that half a pill or one pill were taken for one dose with warm Lophatheri decoction and sugar. The indications and clinical application had developed from the recordings in Xiaoer Yaozheng Zhijue and evolved from pediatrics to ophthalmic otolaryngology， neurology， dermatology， digestion， and respiratory diseases. The main pathogenesis of these diseases is heat in the liver meridian and is treated. The effect of Xieqingwan is "clearing away heat and toxicity， removing fire and relaxing the bowels， and dispersing swelling and relieving pain". It is recommended to use the corresponding preparation methods in the 2020 Edition of Pharmacopoeia of the People's Republic of China. Modern clinical studies are centered around the clinical application of Xieqingwan， which is often modified and used in treating Tourette syndrome， herpes， febrile convulsion， sleepwalking， and insomnia. ConclusionThis paper conducts a thorough textual research of the key information of Xieqingwan， induces its historic evolution， and confirms its key information， so as to provide a reference for the future development of Xieqingwan.