The aryl hydrocarbon receptor (AhR) plays a crucial role in regulating many physiological processes. Activating the AhR-CYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases. Here, a practical in situ cell-based fluorometric assay was constructed to screen AhR-CYP1A1 axis modulators, via functional sensing of CYP1A1 activities in live cells. Firstly, a cell-permeable, isoform-specific enzyme-activable fluorogenic substrate for CYP1A1 was rationally constructed for in-situ visualizing the dynamic changes of CYP1A1 function in living systems, which was subsequently used for discovering the efficacious modulators of the AhR-CYP1A1 axis. Following screening of a compound library, LAC-7 was identified as an efficacious activator of the AhR-CYP1A1 axis, which dose-dependently up-regulated the expression levels of both CYP1A1 and AhR in multiple cell lines. LAC-7 also suppressed macrophage M1 polarization and reduced the levels of inflammatory factors in LPS-induced bone marrow-derived macrophages. Animal tests showed that LAC-7 could significantly mitigate DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice, and markedly reduced the levels of multiple inflammatory factors. Collectively, an optimized fluorometric cell-based assay was devised for in situ functional imaging of CYP1A1 activities in living systems, which strongly facilitated the discovery of efficacious modulators of the AhR-CYP1A1 axis as novel anti-inflammatory agents.

Notum, a negative feedback regulator of the Wnt signaling, has emerged as a promising target for treating glucocorticoid-induced osteoporosis (GIOP). This study showcases an efficient strategy for discovering the anti-Notum constituents from herbal medicines (HMs) as novel anti-GIOP agents. Firstly, a rapid-responding near-infrared fluorogenic substrate for Notum was rationally engineered for high-throughput identifying the anti-Notum HMs. The results showed that Bu-Gu-Zhi (BGZ), a known anti-osteoporosis herb, potently inhibited Notum in a competitive-inhibition manner. To uncover the key anti-Notum constituents in BGZ, an efficient strategy was adapted via integrating biochemical, phytochemical, computational, and pharmacological assays. Among all identified BGZ constituents, three furanocoumarins were validated as strong Notum inhibitors, while 5-methoxypsoralen (5-MP) showed the most potent anti-Notum activity and favorable safety profiles. Mechanistically, 5-MP acted as a competitive inhibitor of Notum via creating strong hydrophobic interactions with Trp128 and Phe268 in the catalytic cavity of Notum. Cellular assays showed that 5-MP remarkably promoted osteoblast differentiation and activated Wnt signaling in dexamethasone (DXMS)-challenged MC3T3-E1 osteoblasts. In dexamethasone-induced osteoporotic mice, 5-MP strongly elevated bone mineral density (BMD) and improved cancellous and cortical bone thickness. Collectively, this study constructs a high-efficient platform for discovering key anti-Notum constituents from HMs, while 5-MP emerges as a promising anti-GIOP agent.

Human carboxylesterase 2A (hCES2A) plays pivotal roles in prodrug activation and hydrolytic metabolism of ester-bearing chemicals. Targeted inhibition of intestinal hCES2A represents a feasible strategy to mitigate irinotecan-triggered gut toxicity (ITGT), but the orally active, selective, and efficacious hCES2A inhibitors are rarely reported. Here, a novel drug-like hCES2A inhibitor was developed via three rounds of structure-based drug design (SBDD) and structural optimization. Initially, donepezil was identified as a moderate hCES2A inhibitor from 2000 US Food and Drug Administration (FDA)-approved drugs. Following two rounds of SBDD and structural optimization, a donepezil derivative (B7) was identified as a strong reversible hCES2A inhibitor. Subsequently, nine B7 carbamates were rationally designed, synthesized and biologically assayed. Among all synthesized carbamates, C3 showed the most potent time-dependent inhibition on hCES2A (IC₅₀ = 0.56 nmol/L), excellent specificity and favorable drug-like properties. C3 could covalently modify the catalytic serine of hCES2A with high selectivity, while this agent also showed favorable safety profiles, high intestinal exposure, and impressive effects for ameliorating ITGT in both human intestinal organoids and tumor-bearing mice. Collectively, this study showcases a rational strategy for developing drug-like and serine-targeting covalent inhibitors against target serine hydrolase(s), while C3 emerges as a promising orally active drug candidate for ameliorating ITGT.

ObjectiveTo study effect of brusatol （BR） on proliferation of human gastric cancer HGC-27 cells and its mechanism. MethodCell counting kit-8 （CCK-8） was used to detect the survival rate of HGC-27 cells at different concentrations of BR. HGC-27 cells were treated with BR （7.5， 15， 30 μmol·L^-1） for 24 h， and then the cell clone formation was analyzed. 2，7-Dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescent probe and lipid peroxidation sensor （C11-BODIPY） were employed to detect the levels of intracellular reactive oxygen species （ROS） and lipid peroxidation， respectively. Real-time polymerase chain reaction （Real-time PCR） and Western blot were performed to detect the messenger ribonucleic acid （mRNA） and protein expressions of solute carrier family 7 member 11 （SLC7A11）， glutathione peroxidase 4 （GPX4）， solute carrier family 40 member 1 （SLC40A1）， transferrin， nuclear factor E₂-related factor 2 （Nrf2） and heme oxygenase-1 （HO-1）， respectively. ResultThe survival rate of HGC-27 cells was decreased with the increase of BR concentration， and the IC₅₀ was 15.34 μmol·L^-1. Compared with the conditions in blank group， the cell clone formation of BR （7.5， 15， 30 μmol·L^-1） groups was inhibited in a dose-dependent manner （P<0.05，P<0.01）， while the levels of intracellular ROS and lipid peroxidation， iron concentration， and lactic dehydrogenase （LDH） leakage were increased in a dose-dependent manner （P<0.05，P<0.01）. Compared with the blank group， the BR （15， 30 μmol·L^-1） groups lowered the mRNA and protein expressions of SLC7A11， GPX4， SLC40A1， Nrf2 and HO-1， while elevated the mRNA and protein expression of TRF in a dose-dependent manner （P<0.05，P<0.01）. ConclusionBR suppressed the proliferation of HGC-27 cells through inducing ferroptosis via inhibiting Nrf2/HO-1 pathway.

Objective:To investigate the expression level of small nucleolar RNA SNORD15A in bone marrow of patients with acute leukemia (AL) and its relationship with clinical characteristics and prognosis of patients.Methods:Bone marrow blood samples of 53 newly treated AL patients and 29 healthy subjects without clinical diagnosis of hematologic diseases or other malignant diseases (control group) at the Affiliated Hospital of Guangdong Medical University from March 2018 to December 2021 were collected. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of SNORD15A in bone marrow blood mononuclear cells of the two groups. The median relative expression of SNORD15A (0.148) was used as the boundary, and AL patients were divided into low expression group (<0.148) and high expression group (≥0.148). The relationship between the expression level of SNORD15A and the clinical characteristics, clinical indicators and overall survival (OS) of AL patients was analyzed. Kaplan-Meier method was used for survival analysis and log-rank test was performed; Cox proportional hazards model was used for univariate and multivariate analyses of OS of patients.Results:The relative expression of SNORD15A was 0.148 (0.012-1.376) in newly treated AL patients and 0.921 (0.513-2.288) in the control group, and the difference was statistically significant ( Z = -6.85, P < 0.01). The differences in SNORD15A relative expression between patients with different prognostic stratification, efficacy and with or without fever and bleeding were statistically significant (all P < 0.05). The differences in platelet count, plateletcrit and albumin levels between SNORD15A low expression group and high expression group were statistically significant (all P < 0.05), and the differences in molecular biology and cytogenetic characteristics were not statistically significant (all P > 0.05). The patients in SNORD15A high expression group had better OS than the low expression group ( P < 0.05). The results of univariate Cox regression analysis showed that SNORD15A was an influencing factor for patients' OS ( HR = 0.063, 95% CI 0.005-0.766, P < 0.05); the results of multivariate Cox regression analysis showed that fatigue ( HR = 4.754, 95% CI 1.014-22.290), fever ( HR = 0.147, 95% CI 0.029-0.746) and hemoglobin ( HR = 0.970, 95% CI 0.944 -0.998) were independent influencing factors for OS (all P < 0.05). Conclusions:SNORD15A is lowly expressed in AL and may be an indicator for disease monitoring and prognostic assessment in AL patients.

Objective: To retrospectively analyze the efficacy and prognostic factors of postoperative intensity modulated radiotherapy for grade Ⅱ gliomas. Methods: Retrospective analysis was conducted on patients with postoperative grade Ⅱ glioma in our hospital from Jan. 2010 to Dec. 2018. The primary endpoint was progression-free survival, and the secondary endpoint was overall survival. Correlative analyses of prognosis by age, gender, initial resection status, the maximum diameter of the lesions, bi-hemisphere, astrocytoma, chemoradiation, adjuvant chemotherapy were conducted. Results: A total of 109 cases with grade Ⅱ glioma were enrolled. The follow-up rate was 91.75%, including 10 cases dead and 27 relapsed. There were 24 cases (88.9%) of in-field failure, and 3 cases (11.1%) of out-field failure. 14 cases of recurrence occurred in 81 cases of total resection group, accounting for 17.3%, and 13 in 28 cases of subtotal resection group, accounting for 46.4%. The recurrence rate in the subtotal resection group was significantly higher than that in the total resection group (χ ²=9.484, P<0.05). The 1-, 2-, 3-, 4- and 5-year progression-free survival rates were 92.5%, 86.0%, 80.6%, 77.5% and 66.8%, respectively. The 2-, 3-, 4- and 5-year overall survival rates were 97.2%, 90.8%, 87.7% and 84.5%, respectively. Multivariate analysis showed that patients with subtotal resection(HR=3.608, P<0.05) and bi-hemisphere(HR=3.183, P<0.05)were significantly correlated with the progression free survival. Conclusions Postoperative intensity modulated radiotherapy for grade Ⅱ gliomas can achieve a better PFS. Recurrence in the radiation field is the main failure mode. Initial resection status and bi-hemisphere of tumor are important influential factors for PFS of grade Ⅱ gliomas patients.