PDIA3 orchestrates effector T cell program by serving as a chaperone to facilitate the non-canonical nuclear import of STAT1 and PKM2.

Dysregulated T cell activation underpins the immunopathology of rheumatoid arthritis (RA), yet the machineries that orchestrate T cell effector program remain incompletely understood. Herein, we leveraged bulk and single-cell RNA sequencing data from RA patients and validated protein disulfide isomerase family A member 3 (PDIA3) as a potential therapeutic target. PDIA3 is remarkably upregulated in pathogenic CD4 T cells derived from RA patients and positively correlates with C-reactive protein level and disease activity score 28. Pharmacological inhibition or genetic ablation of PDIA3 alleviates RA-associated articular pathology and autoimmune responses. Mechanistically, T cell receptor signaling triggers intracellular calcium flux to activate NFAT1, a process that is further potentiated by Wnt5a under RA settings. Activated NFAT1 then directly binds to the Pdia3 promoter to enhance the expression of PDIA3, which complexes with STAT1 or PKM2 to facilitate their nuclear import for transcribing T helper 1 (Th1) and Th17 lineage-related genes, respectively. This non-canonical regulatory mechanism likely occurs under pathological conditions, as PDIA3 could only be highly induced following aberrant external stimuli. Together, our data support that targeting PDIA3 is a vital strategy to mitigate autoimmune diseases, such as RA, in clinical settings.

Preparation of an isorhamnetin phospholipid complex for improving solubility and anti-hyperuricemia activity.

To improve the solubility and anti-hyperuricemia activity of the insoluble natural flavonoid isorhamnetin (ISO), an isorhamnetin phospholipid complex (ISO-PC) was prepared. ISO-PC was prepared through solvent evaporation and its prescription process was optimized. The formation of ISO-PC was verified via multiple characterization methods. Parameters such as drug loading, solubility, octanol-water partition coefficient, stability, and in vivo anti-hyperuricemia activity of ISO-PC were investigated. The complexation efficiency of ISO-PC was 95.1% ± 0.56%. The characterization results confirmed that ISO-PC was bound by intermolecular interactions between ISO and phospholipids. Compared with ISO, the solubility of ISO-PC in water and 1-octanol increased by 122 and 16.5 times, respectively. In addition, the octanol-water partition coefficient decreased to 1.08. Pharmacodynamic studies have reported that ISO-PC has a more significant effect on reducing serum uric acid levels and renal protection. In conclusion, the findings of this study suggested that ISO-PC could be used as a promising formulation to improve the solubility and the anti-hyperuricemia activity of ISO.

Preparation and characterisation of kaempferol composite carrier solid dispersion: evaluation of its application in preventing soybean oil spoilage.

Kaempferol (KPF) can be used as a natural antioxidant and food additive in food processing. However, the poor solubility of KPF limited its bioavailability and application. In order to improve the solubility of KPF, kaempferol composite carrier solid dispersion (KPF-CC-SD) was prepared and the process was optimised. When the ratio of KPF: CA (citric acid): Soluplus reached 1:4:6, the dissolution rate was the highest, and the sample was stable over 12 weeks. The characterisation results indicated that KPF-CC-SD exists in an amorphous form. Peroxidation value and acid value of soybean oil showed that the preservation effect of KPF-CC-SD was better than that of KPF, and the inhibition effect of KPF-CC-SD on acid value was better than that of butylated hydroxytoluene. In conclusion, KPF-CC-SD can change the solubility, crystal form and spatial stability of KPF through the carrier, which has a great application prospect in the field of food preservation.

Association among physical activity, anxiety and oral health status in Chinese university students: A cross-sectional study.

Background: Evidence is limited regarding the relationship among physical activity, anxiety, and oral health in Chinese university students. This cross-sectional investigation aimed to assess the potential relationship between physical activity, anxiety, and oral health conditions among university students in China. Methods: An online questionnaire measuring physical activity, anxiety status, and oral health condition was completed by 1604 university students. The International Physical Activity Questionnaire Short Form (IPAQ-SF) and Generalized Anxiety Disorder-7 (GAD-7) were selected to evaluate physical activity and anxiety, respectively. Oral health condition was assessed through several self-reported variables, including self-reported toothache, gingival bleeding, frequency of tooth brushing, and use of dental floss. Multivariate logistic regression was performed to analyze the underlying relationship between outcome variables. The control variables included age, height, weight, gender, whether only one-child, education level, parental education level, smoking status, drinking habits, and length of sleep. Path analysis was conducted to disentangle the association between physical activity, anxiety, and oral health conditions. Results: Among 1604 university students, 666 (41.5 %) were males and 938 (58.5 %) were females, with an average of 21.9 ± 2.8 years. Only 833 (51.9 %) reported sufficient physical activity, while 684 (42.6 %) of the subjects displayed varying degrees of anxiety. Self-reported gingival bleeding was associated with insufficient physical activity (OR = 1.25; 95%CI: 1.02-1.55), anxiety (OR = 0.45; 95%CI: 0.27-0.74), frequency of tooth brushing (OR = 0.75; 95%CI: 0.60-0.95) and use of dental floss (OR = 0.75; 95%CI: 0.59-0.96), while toothache was not directly influenced by the physical activity and anxiety among university students. Anxiety markedly mediated the relationship between physical activity and oral health conditions. Conclusions: Anxiety was considered a factor associated with the level of physical activity, tooth brushing habits, and self-reported gingival bleeding among university students. Further investigations are required to elucidate whether oral health conditions could be enhanced through the improvement of anxiety and physical activity.

Cystathionine γ-lyase contributes to exacerbation of periodontal destruction in experimental periodontitis under hyperglycemia.

BACKGROUND: Diabetes is one of the major inflammatory comorbidities of periodontitis via 2-way interactions. Cystathionine γ-lyase (CTH) is a pivotal endogenous enzyme synthesizing hydrogen sulfide (H2S), and CTH/H2S is crucially implicated in modulating inflammation in various diseases. This study aimed to explore the potential role of CTH in experimental periodontitis under a hyperglycemic condition. METHODS: CTH-silenced and normal human periodontal ligament cells (hPDLCs) were cultured in a high glucose and Porphyromonas gingivalis lipopolysaccharide (P.g-LPS) condition. The effects of CTH on hPDLCs were assessed by Cell Counting Kit 8 (CCK8), real-time quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA). The model of experimental periodontitis under hyperglycemia was established on both Cth-/- and wild-type (WT) mice, and the extent of periodontal destruction was assessed by micro-CT, histology, RNA-Seq, Western blot, tartrate-resistant acid phosphatase (TRAP) staining and immunostaining. RESULTS: CTH mRNA expression increased in hPDLCs in response to increasing concentration of P.g-LPS stimulation in a high glucose medium. With reference to WT mice, Cth-/- mice with experimental periodontitis under hyperglycemia exhibited reduced bone loss, decreased leukocyte infiltration and hindered osteoclast formation, along with reduced expression of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in periodontal tissue. RNA-seq-enriched altered NF-κB pathway signaling in healthy murine gingiva with experimental periodontitis mice under hyperglycemia. Accordingly, phosphorylation of p65 (P-p65) was alleviated in CTH-silenced hPDLCs, leading to decreased expression of IL6 and TNF. CTH knockdown inhibited activation of nuclear factor kappa-B (NF-κB) pathway and decreased production of proinflammatory cytokines under high glucose and P.g-LPS treatment. CONCLUSION: The present findings suggest the potential of CTH as a therapeutic target for tackling periodontitis in diabetic patients.

Overview of the pharmacokinetics and pharmacodynamics of URAT1 inhibitors for the treatment of hyperuricemia and gout.

INTRODUCTION: Hyperuricemia is a common metabolic disease, which is a risk factor for gouty arthritis and ureteral stones and may also lead to cardiovascular and chronic kidney disease (CDK). Therefore, hyperuricemia should be treated early. Xanthine oxidase inhibitors (XOIs) and uricosuric agents (UAs), which target uric acid, are two types of medications that are used to treat gout and hyperuricemia. XOIs stop the body from producing excessive uric acid, while UAs eliminate it rapidly via the kidneys. Urate transporter 1 (URAT1) belongs to the organic anion transporter family (OAT) and is specifically localized to the apical membrane of the epithelial cells of proximal tubules. Unlike other organic anion transporter family members, URAT1 identifies and transports organic anions that are primarily responsible for urate transport. AREAS COVERED: This article reviews the pharmacokinetics and pharmacodynamics of the existing URAT1 inhibitors to serve as a reference for subsequent drug studies. EXPERT OPINION: The URAT1 inhibitors that are currently used as clinical drugs mainly include dotinurad, benzbromarone, and probenecid. Results indicate that RDEA3170 may be the most promising inhibitor, in addition to SHR4640, URC-102, and MBX-102, which are in the early stages of development.

Bioactivity and Component Analysis of Water Extract of Sophora japonica against Hyperuricemia by Inhibiting Xanthine Oxidase Activity.

Hyperuricemia (HUA) is a metabolic condition caused by excessive production or low excretion of uric acid (UA) in the body. Xanthine oxidase (XOD) is the key enzyme in the process of metabolism purines to generate UA. In this study, the in vitro inhibitory effect of water extract of the flower bud of Sophora japonica (WESJ) on XOD was investigated by ultraviolet spectrophotometry. A mice model of HUA was constructed to explore the effect of WESJ on UA levels and the mechanism of action on renal function. Based on Box-Behnken design, the optimal extraction process of WESJ was determined to extract Sophora japonica twice with 8 times of water, 0.5 h each time. Pharmacological results showed that low, medium, and high doses of WESJ (200, 400, 600 mg/kg) could significantly reduce serum UA level, inhibit the activity of XOD in blood and liver, and have a protective effect on kidney damage caused by high UA. Through UPLC-Q-TOF-MS/MS analysis, 214 compounds were identified in WESJ, including flavonoids, polyphenols, triterpenoids, organic acids, and others. The rat serum of WESJ was analyzed, and 23 prototype components entering the blood were identified, including 15 flavonoids and polyphenols, which may be the main bioactive components. In conclusion, flavonoids and polyphenols in WESJ may reduce the level of UA and alleviate kidney damage by inhibiting the activity of XOD. WESJ is expected to be used as a plant-based food and dietary supplement for the treatment of HUA.

A brief review of urate transporter 1 (URAT1) inhibitors for the treatment of hyperuricemia and gout: Current therapeutic options and potential applications.

Hyperuricemia is a common metabolic condition, cause by increased levels of serum urate (SUA). Reduced excretion of uric acid is reported as the key factor of primary hyperuricemia, accounting for approximately 90% of the cases. Urate transporter 1 (URAT1) is a major protein involved in uric acid reabsorption (about 90%). Therefore, URAT1 inhibitors are considered to be a highly effective and promising class of uricosuric agents for treating hyperuricemia. This review summarizes the development of URAT1 inhibitors for the treatment of hyperuricemia, including approved URAT1 inhibitors, URAT1 inhibitors under development in clinical trials, substances with URAT1 inhibitory effects from derivatives and natural products, and conventional drugs with new uses. This review provides new ideas regarding research on URAT1 inhibitors by introducing the structure, properties, and side effects of chemical drugs, as well as the sources and categories of natural drugs. We also discuss new mechanisms of classic drugs, which may provide guidance to many practicing clinicians. The research and discovery of new inhibitors remain in full swing, and tremendous developments are expected in the field.

Quantitation of vistusertib by UHPLC-MS/MS in rat plasma and its application to a pharmacokinetic study.

Aim: The present study aimed to develop a UHPLC-MS/MS method for determination of vistusertib in biological matrix, and to describe the pharmacokinetic behavior of vistusertib in SD rats. Methodology & results: After protein precipitation with acetone and acetonitrile (1:1), the chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18 column and detected with a SCIEX QTRAP 4500 mass spectrometer under positive ionization mode. The developed UHPLC-MS/MS method showed an excellent linearity within the range of 1.0-3000 ng/ml with good accuracy and precision. Vistusertib showed a rapid absorption and reached the maximum concentration of 3532.2 ± 678.0 ng/ml 20-30 min after oral administration in Sprague-Dawley rats. Conclusion: The established analytical method was fast, sensitive and robust, and successfully applied to describe the pharmacokinetic behavior of vistusertib following an oral administration in rats.

Isorhamnetin, the xanthine oxidase inhibitor from Sophora japonica, ameliorates uric acid levels and renal function in hyperuricemic mice.

Hyperuricemia is a metabolic condition closely linked to xanthine oxidase (XOD) function, which is involved in the production of uric acid (UA). In this study, XOD was used as a target to construct an in vitro and in vivo activity screening and verification system. The XOD inhibition ability of the main components from the water extract of Sophorae Flos (WSF), an unopened dry flower bud of Sophora japonica, was screened by HPLC. Isorhamnetin (IRh) was identified as a major flavonoid XOD inhibitor from WSF, and we characterized its effects and potential mechanism in ameliorating UA levels and renal function in hyperuricemia model mice. Hyperuricemia was induced by oral administration of potassium oxonate (PO) and hypoxanthine to mice for 7 days. The biochemical index results showed that treatments with low, medium, and high doses of IRh (50, 100, and 150 mg kg-1) significantly reduced serum UA levels and inhibited XOD activity in serum and in the liver. Additionally, IRh effectively decreased the levels of serum creatinine and blood urea nitrogen, suggesting that it possessed nephroprotective effects in hyperuricemic mice. Furthermore, histopathological results showed that nuclear lesions and renal tubule dilatation in the kidneys of IRh-treated hyperuricemic mice were reduced, suggesting that IRh may alleviate renal injury. Molecular docking results showed that IRh combined well with XOD and is an effective XOD inhibitor. In conclusion, IRh from Sophora japonica may reduce the UA levels and alleviate renal injury by inhibiting XOD activity. It potentially functions as a therapeutic drug and dietary supplement to treat hyperuricemia.

Genomic and clinical characteristics of MET exon14 alterations in a large cohort of Chinese cancer patients revealed distinct features and a novel resistance mechanism for crizotinib.

Background: Alterations in MET exon 14 (METex14) and its flanking intronic regions have been identified in a variety of cancers. Patients with METex14 alterations often benefit from MET inhibitors such as crizotinib. Given the unique mutation profiles of Chinese lung cancer patients, it is necessary to investigate the prevalence of METex14 alterations in a large cohort of cancer patients. Patients and methods: Cases carrying METex14 alterations were screened from 26,391 Chinese cancer patients by next-generation sequencing (NGS), and the clinicopathologic and molecular characteristics were reviewed. Results: Compared to Western population (~3%), the frequency of METex14 alterations is much lower in Chinese cancer patients (0.7%, n=184) and lung cancer patients (1.1%, n=175). Seventy-eight distinct METex14 alterations, including several novel alteration types were detected. Concurrent MET copy gain and non-exon14 MET mutations were also found. EGFR copy gain (11%) and mutations (8%), KRAS (5%) and PIK3CA (5%), appeared in a mutually exclusive pattern. Female patients contain much less TP53 mutations than male patients (65% vs. 24%, FDR = 0.01). Co-amplification of CDK4 and MDM2, CDK6 and EGFR were identified, which indicated cell cycle dysregulation and EGFR alteration are important co-occurring features in patients with METex 14 alteration. Of 9 tissue specimens having PD-L1 immunohistochemistry (IHC) results, 5 of them (55.5%) were found PD-L1 positive, which is comparable to other types of tumor. In 14 crizotinib-treated patients, the median progression free survival (mPFS) was 7 months. Upon resistance to crizotinib, two patients acquired secondary mutations in MET and one patient acquired BRAF p.K601E that can be a novel resistance mechanism. Conclusion: Chinese cancer patients have a relatively lower frequency of METex14 alterations compared to Western patients. Patients with METex14 alterations showed distinct molecular characteristics and the representative case study showed responses to MET tyrosine kinase inhibitor (TKI).

Icariin Inhibits Intestinal Inflammation of DSS-Induced Colitis Mice Through Modulating Intestinal Flora Abundance and Modulating p-p65/p65 Molecule.

BACKGROUND: Ulcerative colitis, as a kind of inflammatory bowel disease (IBD) is characterized by abdominal pain. This study aimed to investigate the effect of icariin (ICA) on the intestinal microflora of colitis mice. METHODS: Fifteen female C57BL/6 mice were randomly divided into the Control group, dextran sodium sulfate (DSS)-induced colitis (DSS) group, and ICA treatment (DSS+ICA) group. The severity of inflammation in DSS-induced colitis mice was evaluated using disease activity scoring (considering weight-loss percentage, stool-shape change, and stool-bleeding scoring). Pathological changes of mice intestinal tract were evaluated using hematoxylin-eosin (HE) staining. Serum levels of TNF-α and IL-6 were detected with enzyme-linked immunosorbent assay. Expressions of p65 and p-p65 (p-p65/p65 ratio) were analyzed using Western blot assay. 16S rDNA sequencing was used to analyze the abundance and composition of intestinal microflora. RESULTS: Compared with DSS group, ICA significantly improved disease activity (P < .05) and reduced inflammatory damage of colon tissues (P < .05) in DSS-induced colitis mice. Compared with the DSS group, mice in the ICA group demonstrated significant weight and colon length (P < .05). ICA significantly inhibited expressions of IL-6 and TNF-α compared to the DSS group (P < .05). p-p65/ p65 ratio in the DSS + ICA group was remarkably enhanced compared to the DSS group (P < .05). ICA significantly reduced the proportion and activity of Bacteroides, Helicobacteraceae, Turicibacter, and significantly increased that of beneficial microflora (Lactobacillus, Lachnospiraceae, Akkermansia), so as improved damages of colon tissues. CONCLUSION: ICA can improve intestinal flora abundance and composition of DSS-induced colitis mice, and inhibit tissue damage and inflammatory response through modulating the p-p65/p65 expression.