ObjectiveTo investigate the effects of Yishen Juanbi pills （YSJB）-containing bone marrow fluid on the migration and differentiation phenotypes of CD4⁺T lymphocytes based on the stromal cell-derived factor-1/chemokine receptor 4 （SDF-1/CXCR4） signaling pathway. MethodsPrimary CD4⁺T lymphocytes were isolated from mice using magnetic bead separation and identified for purity by flow cytometry. A CD4⁺T lymphocyte culture system was then established to observe the effects of SDF-1 on CD4⁺T-cell migration and differentiation. On this basis， the experimental groups included the Sham group， the ovariectomy （OVX） group， the Sham+collagen-induced arthritis （CIA） group， the OVX+CIA group， the Sham+CIA+YSJB group （2.16 g·kg^-1）， the OVX+CIA+YSJB group （2.16 g·kg^-1）， and the OVX+CIA+methotrexate （MTX） group （1.5 mg·kg^-1）. Bone marrow fluid from each group was prepared according to previous methods and added to the CD4⁺ T-cell culture system at 5% （v/v）. Transwell assays were used to examine CD4⁺T-cell migration in each group. Real-time PCR was used to measure the mRNA expression levels of interleukin （IL）-17， tumor necrosis factor-α （TNF-α）， retinoic-acid-related orphan receptor γt （RORγt）， IL-10， transforming growth factor-β （TGF-β）， forkhead box P3 （FoxP3）， CXCR4， phosphoinositide 3-kinase （PI3K）， and protein kinase B （Akt）. Western blot was used to detect the expression of helper T （Th）17/regulatory T （Treg） cell signature factors （RORγt， FoxP3）， CXCR4， PI3K， phosphorylated （p）-PI3K， Akt， and p-Akt. In a separate set of experiments， cells were divided into the Sham group， OVX+CIA group， OVX+CIA+CXCR4 antagonist AMD3100 group， and OVX+CIA+YSJB+AMD3100 group to observe changes in the above indicators following AMD3100 intervention. ResultsCompared with the Sham group， the number of migrated cells in the lower chamber was significantly increased in the Sham+CIA and OVX+CIA groups （P<0.05， P<0.01）. The mRNA expression of RORγt， IL-17， TNF-α， CXCR4， PI3K， and Akt was significantly upregulated， whereas FoxP3， IL-10， and TGF-β mRNA expression was significantly decreased （P<0.05， P<0.01）. Protein expression of RORγt， CXCR4， p-PI3K/PI3K， and p-Akt/Akt was significantly increased， while FoxP3 protein expression was markedly decreased （P<0.05， P<0.01）. Compared with the OVX+CIA group， the OVX+CIA+YSJB group and OVX+CIA+MTX group showed significantly reduced migration （P<0.05）， mRNA expression of RORγt， IL-17， TNF-α， CXCR4， PI3K， and Akt was also significantly decreased， while FoxP3， IL-10， and TGF-β mRNA expression was significantly increased （P<0.05， P<0.01）. RORγt protein expression was significantly downregulated， and FoxP3 protein expression markedly upregulated （P<0.05）. In the OVX+CIA+YSJB group， CXCR4， p-PI3K/PI3K， and p-Akt/Akt protein expression was significantly decreased （P<0.05）. Compared with the OVX+CIA group， RORγt， CXCR4， PI3K， and Akt mRNA expression in CD4⁺T cells was significantly decreased in the OVX+CIA+AMD3100 group and the OVX+CIA+YSJB+AMD3100 group， while FoxP3 mRNA and protein expression was significantly upregulated （P<0.05， P<0.01）. RORγt， CXCR4， p-PI3K/PI3K， and p-Akt/Akt protein expression was also markedly decreased （P<0.05， P<0.01）. Compared with the OVX+CIA+AMD3100 group， the OVX+CIA+YSJB+AMD3100 group showed significantly decreased RORγt and Akt mRNA expression （P<0.05） and significantly lower p-Akt/Akt protein expression （P<0.05）. ConclusionYSJB-containing bone marrow fluid suppresses CD4⁺T-cell migration and regulates Th17/Treg balance by downregulating Th17-associated signature factors and upregulating Treg-associated signature factors through inhibition of the SDF-1/CXCR4 signaling pathway and PI3K/Akt signaling pathway. The SDF-1/CXCR4 signaling pathway is one of the targets through which YSJB inhibits CD4⁺T-cell differentiation.

In recent years， hyperuricemia （HUA） has shown a rapidly increasing incidence and tends to occur in increasingly young people， with a wide range of cardiac， renal， joint， and cancerous hazards and all-cause mortality associations. Western medicine treatment has limitations such as large liver and kidney damage， medication restriction， and easy recurrence. The intestine is the major extra-renal excretion pathway for uric acid （UA）， and the intestinal microecology can be regulated to promote UA degradation. It offers great potential to develop UA-lowering strategies that target the intestinal microecology， which are promising to provide safer and more effective therapeutic approaches. Traditional Chinese medicine （TCM） can treat HUA via multiple targets and multiple pathways from a holistic view， with low toxicity and side effects. Studies have shown that intestinal microecology is a crucial target for TCM in the treatment of HUA. However， its specific mechanism of action has not been fully elucidated. Focusing on the key role of intestinal microecology in HUA， this review explores the relationship between intestinal microecology and HUA in terms of intestinal flora， intestinal metabolites， intestinal UA transporters， and intestinal barriers. Furthermore， we summarize the research progress in TCM treatment of HUA by targeting the intestinal microecology， with the aim of providing references for the development of TCM intervention strategies for HUA and the direction of future research.

Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response. Stem cell therapy can be a promising treatment strategy for periodontitis, but the relevant mechanism is complicated. This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs) for the treatment of periodontitis. The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure. Human gingival fibroblasts (HGFs) were exposed to 5 μg/mL lipopolysaccharide (LPS) for 24 h to establish a cell injury model. When treated with hESC-MSCs or mitochondria derived from hESC-MSCs, HGFs showed reduced expression of inflammatory genes, increased adenosine triphosphate (ATP) level, decreased reactive oxygen species (ROS) production, and enhanced mitochondrial function compared to the control. The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%. Besides, a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression, as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues. In conclusion, our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs, suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.
Humans ; Gingiva/cytology* ; Fibroblasts/metabolism* ; Mitochondria/physiology* ; Mesenchymal Stem Cells/cytology* ; Animals ; Periodontitis/therapy* ; Mice ; Reactive Oxygen Species/metabolism* ; Inflammation ; Lipopolysaccharides ; Human Embryonic Stem Cells/cytology* ; Cells, Cultured ; Adenosine Triphosphate/metabolism* ; Male

Despite therapy with potent antiviral agents, chronic hepatitis B (CHB) patients remain at high risk of hepatocellular carcinoma (HCC). While metabolites have been rediscovered as active drivers of biological processes including carcinogenesis, the specific metabolites modulating HCC risk in CHB patients are largely unknown. Here, we demonstrate that baseline plasma from CHB patients who later developed HCC during follow-up exhibits growth-promoting properties in a case-control design nested within a large-scale, prospective cohort. Metabolomics analysis reveals a reduction in long-chain acylcarnitines (LCACs) in the baseline plasma of patients with HCC development. LCACs preferentially inhibit the proliferation of HCC cells in vitro at a physiological concentration and prevent the occurrence of HCC in vivo without hepatorenal toxicity. Uptake and metabolism of circulating LCACs increase the intracellular level of acetyl coenzyme A, which upregulates histone H3 Lys14 acetylation at the promoter region of KLF6 gene and thereby activates KLF6/p21 pathway. Indeed, blocking LCAC metabolism attenuates the difference in KLF6/p21 expression induced by baseline plasma of HCC/non-HCC patients. The deficiency of circulating LCACs represents a driver of HCC in CHB patients with viral control. These insights provide a promising direction for developing therapeutic strategies to reduce HCC risk further in the antiviral era.

Objective: To analyze the physical and chemical properties of human gastric factor by bioinformatics method , and to express and purify the protein. Methods: Protein online analysis software was used to predict the physical and chemical properties of gastric intrinsic factor and analyze their hydrophilicity and hydrophobicity. The online tool was used to predict and analyze the human gastric factor signal peptide and its subcellular location . The eukaryotic expression recombinant plasmid pcDNA3 . 1 ( + ) _human GIF_His tag was constructed , and the gastric intrinsic factor was purified by nickel column after expression in HEK293F cells . The purity and activity of purified gastric intrinsic factor were verified by SDS_PAGE , Western blot and indirect ELISA . Results: Gastric factor contained 417 amino acids and was a hydrophilic acid stable protein . It was a secreted protein with Sec original signal peptide . pcDNA3 . 1 ( + ) _human GIF_His tag recombinant plasmid was successfully constructed and soluble expression was obtained in HEK293F cell expression system. Conclusion The eukaryotic source of human gastric intrinsic factor is successfully prepared , and the bioinformatics results show that the protein is a hydrophilic acid stable secreted protein , laying a foundation for the subsequent use of this protein as an immunogen and protein calibrator to construct an immunoassay for gastric factor.

ObjectiveBased on transcriptomics， to explore the mechanism of Hei Xiaoyaosan regulating the phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway to improve the learning and memory ability of insomnia rats with liver depression syndrome. MethodsSixty 8-week-old male SD rats were randomly divided into the blank group， model group， eszopiclone group （0.09 mg·kg^-1）， and low， medium， and high dose groups of Hei Xiaoyaosan （3.82， 7.65， 15.30 g·kg^-1）， with ten rats in each group. Except for the blank group， the other groups were induced insomnia rat model with liver depression by chronic restraint， tail clamping stimulation and intraperitoneal injection of p-chlorophenylalanine （PCPA）. Each treatment group received intragastric administration according to the specified dosage， once a day for 14 consecutive days. The pentobarbital sodium cooperative sleep test， open field test， and Morris water maze test were used to test the sleep quality， depressive-like behavior， and learning and memory abilities of rats. Additionally， enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of 5-hydroxytryptamine （5-HT）， γ-aminobutyric acid （GABA）， brain-derived neurotrophic factor （BDNF）， glial cell line-derived neurotrophic factor （GDNF） and nitric oxide （NO） in hippocampus. Hematoxylin-eosin （HE） staining was performed to observe pathological changes of the hippocampal tissue， while terminal deoxynucleotidyl transferase deoxyuridine triphosphate （dUTP） nick end labeling （TUNEL） was used to evaluate apoptosis of hippocampal neurons. Transcriptomic sequencing technology was employed to identify differentially expressed genes in hippocampus between the model group and the blank group， as well as between the medium-dose group of Hei Xiaoyaosan and the model group. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis were performed on the intersecting genes. Subsequently， the enriched key genes and signaling pathways were analyzed and verified. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was utilized to assess the mRNA expression levels of phosphatase and tensin homolog （PTEN）， B-cell lymphoma-2 （Bcl-2）-like protein 11 （BCL2L11）， and mitogen-activated protein kinase 1 （MAPK1） in hippocampus， and Western blot was employed to evaluate the protein expressions of PI3K， phosphorylation （p）-PI3K， Akt， p-Akt， Bcl-2， Bcl-2-associated X protein （Bax）， and cleaved Caspase-3 in the same tissue. ResultsCompared with the blank group， the model group exhibited a reduction in body weight， an increase in sleep latency， and a decrease in sleep duration （P<0.01）. Additionally， rats showed obvious depression-like behavior， and their learning and memory abilities decreased. Furthermore， the contents of 5-HT， GABA， NO， BDNF and GDNF in hippocampus decreased （P<0.01）. Histological examination revealed a disorganized cell arrangement in the CA1 region of the hippocampus， characterized by irregular cell shapes， a reduced cell count， deeply stained and pyknotic nuclei， increased vacuolar degeneration， and an elevated apoptosis rate （P<0.01）. Compared with the model group， the body weight of the high and medium dose groups of Hei Xiaoyaosan increased， the sleep latency shortened and the sleep time prolonged （P<0.05， P<0.01）. Additionally， depression-like behavior and learning and memory abilities of rats were significantly improved， the levels of 5-HT， GABA， NO， BDNF and GDNF in the hippocampus increased （P<0.05， P<0.01）. These interventions also ameliorated pathological damage in the hippocampal CA1 area and reduced the apoptosis of hippocampal neurons （P<0.01）. Transcriptomic sequencing results indicated that Hei Xiaoyaosan might exert a therapeutic effect by regulating PI3K/Akt pathway through key mRNAs such as PTEN， BCL2L11， and MAPK1. The roles of these key mRNAs and proteins within PI3K/Akt pathway were further validated. In comparison to the blank group， the expression levels of PTEN， BCL2L11 and MAPK1 mRNA in the hippocampus of rats in the model group were increased （P<0.01）， while the protein expression levels of p-PI3K， p-Akt and Bcl-2 were decreased （P<0.01）， and the protein expression levels of PTEN， Bax and cleaved Caspase-3 were increased （P<0.01）. Compared with the model group， the high-dose and medium-dose groups of Hei Xiaoyaosan could down-regulate the expressions of PTEN， BCL2L11 and MAPK1 mRNAs （P<0.01）， up-regulate the expressions of p-PI3K， p-Akt and Bcl-2 proteins （P<0.01）， and down-regulate the protein expressions of PTEN， Bax and cleaved Caspase-3 （P<0.05， P<0.01）. ConclusionHei Xiaoyaosan may regulate PI3K/Akt signaling pathway by down-regulating expressions of key genes such as PTEN， BCL2L11 and MAPK1， and thus improve the learning and memory abilities of insomnia rats with liver depression syndrome.

Objective To investigate the relationship between serum C-C motif ligand 23 (CCL23),Stanniocalcin-1 (STC1) levels and prognosis in patients with severe hypertensive intracerebral hemorrhage (HICH). Methods A total of 122 severe HICH patients who visited the Department of Neurosurgery,Hohhot First Hospital from March 2021 to March 2023 were regarded as the study subjects (HICH group),122 patients with mild HICH during the same period (mild group) and 122 healthy individuals who underwent physical examinations were considered healthy. HICH patients were separated into survival group(n=94) and death group(n=28)based on prognosis. ELISA was applied to detect serum levels of CCL23 and STC1. Spearson on method was used to analyze correlations and multivariate COX regression was used to investigate the influencing factors of prognosis in HICH patients,and ROC curve was applied to analyze the predictive value of serum CCL23 and STC1 levels for the prognosis. Kaplan-Meier was applied to analyze the relationship between serum CCL23,STC1 levels and clinical outcomes. Results Serum CCL23(53.32±10.85pg/ml,78.49±11.21pg/ml,112.47±11.53pg/ml)and STC1 (15.12±2.63ng/ml,19.07±2.58ng/ml,22.15±2.75ng/ml)levels in the healthy group,mild disease group and HICH group were increased successively,and the differences was statistically significant (F=856.967,215.043,all P<0.05). The serum levels of CCL23 (108.02±13.51pg/ml) and STC1 (21.06±3.28ng/ml) in the survival group were lower than those in the death group(127.41±13.55 pg/ml,25.83±3.23 ng/ml),the Glasgow coma (GCS) score (8.95±0.92 ) of the survival group was higher than that of the death group(7.61±0.77),and the differences were statistically significant (t=6.663,6.810,7.005,all P<0.001). The serum levels of CCL23 and STC1 were negatively correlated with GCS score (r=-0.481,-0.426,all P<0.001). CCL23[OR(95％CI):1.240(1.091～1.409)],STC[OR(95％CI):1.754(1.215～2.533)]and GCS[OR(95％CI):0.087(0.020～0.382)]score were the influencing factors for poor prognosis in HICH patients . The AUC(95％CI) of CCL23 combined STC1 in the prediction of the prognosis of HICH patients was 0.939 (0.880～0.974) which was higher than that of single diagnosis (Z=1.974,2.040,P=0.048,0.041),the sensitivity and specificity of combined diagnosis were 85.71％ and 94.68％,respectively. The 6-month follow-up survival rate of patients with high expression of CCL23 and STC1 (51.06％ vs 93.33％,56.86％ vs 91.55％) was lower than that of patients with low expression of CCL23 and STC1,and the differences were statistically signrficant (Log rank x2=34.777,23.781,all P<0.05). Conclusion The serum levels of CCL23 and STC1 are high in severe HICH patients,which are closely related to their prognosis. High expression of CCL23 and STC1 may indicate poor clinical outcomes in patients.

Objective:To observe the clinical efficacy and effects on emotional state, anorectal physiological function, serum inflammation factors and neurotransmitters of repetitive transcranial magnetic stimulation (rTMS) on functional anorectal pain (FAP) patients, and to explore the potential therapeutic mechanisms.Methods:From September 1, 2022 to December 31, 2023, a total of 50 FAP patients who were admitted to Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine were enrolled in this study. The patients were randomly divided into the treatment group (20 cases) and the control group (20 cases) according to a random number table and relevant exclusion criteria. The treatment group received rTMS treatment and the control group received sham rTMS treatment. The Hamilton anxiety scale (HAMA) score, Hamilton depression scale (HAMD) score, visual analogue scale (VAS) score, high-resolution anorectal manometry data (anal resting pressure, anal squeeze pressure, initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold), and the levels of serum inflammatory factors (interleukin(IL)-4, IL-8, tumor necrosis factor-α) and 5-hydroxytryptamin(5-HT) were recorded before and after treatment. Independent sample t-test, paired t-test, Mann-Whitney U test and Wilcoxon signed-rank test were used for statistical analysis. Results:The VAS, HAMA, and HAMD scores of the treatment group after treatment were lower than those before treatment (3.85±2.23 vs. 6.85±1.98, 4.40±3.39 vs. 8.75±6.60, and 7.10±6.56 vs. 12.85±7.20), and were also lower than those of the control group after treatment(6.50±1.76, 8.20±6.65, 12.10±6.80), and the differences were statistically significant ( t=5.68, 4.72, 6.06; -4.17, -2.27, -2.37; P<0.001, <0.001, <0.001; <0.001, =0.028, and =0.023). The initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold of the treatment group after treatment were higher than those before treatment(30.00(30.00, 46.00) mmHg (1 mmHg=0.133 kPa) vs. 23.00(18.50, 29.00) mmHg, 50.00(44.50, 60.00) mmHg vs. 37.00(30.75, 51.50) mmHg, (74.30±16.02) mmHg vs. (63.70±22.21) mmHg, 119.00(100.00, 148.00) mmHg vs. 98.00 (69.50, 153.00) mmHg), and the tolerance shreshold of the treatment group after treatment was higher than that of the control group after treatment(119.00 (100.00, 148.00) mmHg vs. 102.00(84.50, 111.50) mmHg), and the differences were statistically significant ( Z=–3.14 and –2.86, t=-4.02, Z=-2.84 and -2.11; P=0.002, 0.004, 0.001, 0.004, and 0.035). Additionally, the 5-HT level of the treatment group after treatment was higher than that before treatment (1 549.41 (1 320.21, 1 640.03) μg/L vs. 1 081.52(874.36, 1 626.79) μg/L), and the difference was statistically significant ( Z=-2.88, P=0.004). Conclusion:The rTMS treatment can effectively relieve the pain, anxiety and depression, improve visceral sensitivity, and influence the neurotransmitter level of brain-gut axis in FAP patients.

Objective To analyze the value of MRI in evaluating the treatment efficacy of neoadjuvant chemotherapy(NAC)in breast cancer patients.Methods The clinical data of 130 patients with breast cancer were analyzed retrospectively.All patients under-went 4-8 cycles of NAC,with MRI examinations was performed before and after NAC treatment.Based on postoperative response eval-uation criteria in solid tumors(RECIST),the patients were categorized into effective and ineffective groups.The MRI related param-eters,early enhancement parameters and peak enhancement parameters before and after NAC were observed to evaluate the efficacy of MRI in evaluating the treatment efficacy of NAC in breast cancer.Results The sensitivity,specificity,and accuracy of MRI in evalua-ting the treatment efficacy of NAC in breast cancer were 96.74％,89.47％,and 94.62％,respectively.The Kappa value for consis-tency analysis between MRI and postoperative pathology reached 0.869,indicating good agreement.Receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)for MRI in predicting the treatment efficacy of NAC in breast cancer was 0.935[95％ confidence interval(CI)0.831-0.981].Significant differences were observed between the effective and ineffec-tive groups in early and peak enhancement parameters before and after NAC(P＜0.05).Conclusion MRI can effectively assesses the treatment efficacy of NAC in breast cancer patients,demonstrating high value in clinical applications.

Rheumatic diseases,as a complex class of chronic immune-mediated disorders,involve interplay among genetics,environment,and immunity in their pathogenesis,which remains incompletely elucidated to date.In recent years,with the advancement of epigenetic research,particularly in the field of RNA modifications,the role of m6 A methylation in rheumatic diseases has increasingly garnered attention.Traditional Chinese medicine(TCM),as the indigenous medical system of China,has accumulated extensive experience in the treatment of rheumatic diseases.This article systematically reviewed the research progress of TCM in treating rheumatic diseases such as rheumatoid arthritis,systemic lupus erythematosus,Sj?gren's syndrome,ankylosing spondylitis,and osteoarthritis,as well as improving bone metabolism abnormalities closely related to rheumatic diseases,through regulating m6 A methylation.The aim is to provide novel insights into the treatment of rheumatic diseases with TCM and to offer theoretical support for the development of related functional drugs.