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.

Objectives:To explore the impact of intensive lipid-lowering strategy on short-term prognosis of acute coronary syndrome(ACS)patients with multi-vessel disease. Methods:A total of 136 ACS patients with multi-vessel disease who received coronary stenting at Zhongnan Hospital of Wuhan University from August 2019 to November 2020 were enrolled in this study.Patients were divided into intensive lipid-lowering group(control low density lipoprotein cholesterol[LDL-C]below 1.0 mmol/L within 3 months,and continuously meet the standards within 12 months,n=69)or standard lipid-lowering group(gradually control LDL-C below 1.4 mmol/L within one year,n=67).The total cholesterol(TC),triglycerides(TG),LDL-C,high-density lipoprotein cholesterol(HDL-C),and lipoprotein(a)(Lp[a])data were collected.Incidence of major adverse cardiovascular events(MACE,including cardiac death,myocardial infarction,target vessel revascularization and stroke)were observed during 12 months of follow up. Results:The baseline data of the intensive lipid-lowering group and the standard lipid-lowering group were consistent before intervention.At the timeline of enrollment,there was no statistically significant difference in the blood lipid profiles(including TC,TG,LDL-C,HDL-C)between the two groups.After 3-months,patients in the intensive lipid-lowering group experienced significantly lower TC,TG,LDL-C and Lp(a)compared with baseline values(all P<0.05),while HDL-C remained unchanged(P>0.05).The standard lipid-lowering group showed a significant decrease in TC and LDL-C compared with baseline values(both P<0.05).The TC and LDL-C levels were significantly lower in the intensive lipid-lowering group than in the standard lipid-lowering group at 3/6/12 months follow up after discharge(all P<0.01).At 12 months follow-up,Kaplan-Meier survival analysis showed that the incidence of MACE was significantly lower in the intensive lipid-lowering group than in the standard lipid-lowering group(2.90%vs.14.93%,χ2=6.090,P=0.014).Multiple Cox regression analysis revealed that the intensive lipid-lowering strategy significantly reduced the risk of MACE compared with the standard lipid-lowering strategy(HR=0.177,95%CI:0.037-0.838,P=0.029). Conclusions:Our data show that intensive lipid-lowering strategy may probably reduce the incidence of short-term MACE in ASC patients with multi-vessel disease.Large-scale prospective multi-center studies are needed to further validate these results.

BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the pathophysiology of sepsis, but the exact mechanism remains debatable. In this study, we investigated the associations among the serum levels of PAI-1, the incidence of 4G/5G promoter PAI-1 gene polymorphisms, immunological indicators, and clinical outcomes in septic patients. METHODS: A total of 181 patients aged 18-80 years with sepsis between November 2016 and August 2018 in the intensive care unit in the Xinhua Hospital were recruited in this retrospective study, with 28-day mortality as the primary outcome. The initial serum level of PAI-1 and the presence of rs1799768 single nucleotide polymorphisms (SNPs) were examined. Univariate logistic regression and multivariate analyses were performed to determine the factors associated with different genotypes of PAI-1, serum level of PAI-1, and 28-day mortality. RESULTS: The logistic analysis suggested that a high serum level of PAI-1 was associated with the rs1799768 SNP of PAI-1 (4G/4G and 4G/5G) (Odds ratio [OR]: 2.49; 95% confidence interval [CI]: 1.09, 5.68). Furthermore, a high serum level of PAI-1 strongly influenced 28-day mortality (OR 3.36; 95% CI 1.51, 7.49). The expression and activation of neutrophils (OR 0.96; 95% CI 0.93, 0.99), as well as the changes in the expression patterns of cytokines and chemokine-associated neutrophils (OR: 1.00; 95% CI: 1.00, 1.00), were both regulated by the genotype of PAI-1. CONCLUSIONS Genetic polymorphisms of PAI-1 can influence the serum levels of PAI-1, which might contribute to mortality by affecting neutrophil activity. Thus, patients with severe sepsis might clinically benefit from enhanced neutrophil clearance and the resolution of inflammation via the regulation of PAI-1 expression and activity.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Humans ; Middle Aged ; Young Adult ; Genotype ; Neutrophils ; Plasminogen Activator Inhibitor 1/genetics* ; Polymorphism, Single Nucleotide/genetics* ; Retrospective Studies ; Sepsis/genetics*

Objective To explore the correlation between changes of gray matter volume and related cognitive impairment domains in patients with cognitive impairment of cerebral small vessel disease(CSVD)based on 7.0T magnetic resonance imaging(MRI)and voxel-based morphometry(VBM).Methods All subjects were recruited from the study on Correlation between Cerebral Deep Medullary Vein Morphology and Cognitive Impairment due to Cerebral Small Vessel Disease(registration No.:ChiCTR2100045136)from September 2021 to June 2023.We retrospectively enrolled CSVD patients with cognitive impairment as CSVD group and healthy controls with matched age,gender and education level as control group according to inclusion and exclusion criteria.Montreal cognitive assessment(MoCA)scale(Beijing version)score＜26 was divided into cognitive impairment.All subjects was assessed with MoCA,digit span test(DST),digit symbol substitution test(DSST),trail making test-A(TMT-A),verbal fluency test(VFT),Boston naming test(BNT)and auditory verbal learning test(AVLT).All subjects underwent 7.0T brain MRI scan to acquire T1-weighted three-dimensional magnetization prepared 2 rapid gradient echo(T1WI-MP2RAGE)for VBM analysis.General data and above cognitive function scores were compared between 2 groups.VBM analysis was used to compare the gray matter volume(GMV)between 2 groups and get mean GMV of significant brain regions of CSVD to explore the correlation between regions and cognitive function scores.Results(1)There were 18 individuals in control group,aged 55-70 years,and 19 individuals in CSVD group,aged 57-75 years.There was no significant difference in age,gender,education,body mass index,history of coronary heart disease,history of hyperlipidemia,smoking,drinking,total cholesterol,triglyceride,low density lipoprotein and high density lipoprotein between the two groups(all P＞0.05).But the proportion of hypertension and diabetes history in the CSVD group was higher than control group,and there were significant differences between the two groups(12/19 vs.5/18,7/19 vs.0;all P＜0.05).(2)The scores of MoCA scale(22.0[20.0,23.0]vs.27.0[26.0,28.0],Z=-5.242),DSST(18±9 vs.40±4,t=5.212),DST(10.6±2.5 vs.13.9±2.0,t=4.364),VFT(38±11 vs.47±8,t=3.224),AVLT-immediate memory(13±3 vs.21±4,t=6.877),AVLT-short delay recall(3.4±2.5 vs.6.9±2.2,t=4.555)and BNT(22.7±3.6 vs.27.0±2.1,t=4.357)in CSVD group were lower than those in the control group.The time taken to complete TMT-A in CSVD group was longer than the control group(93.00[76.04,125.69]s vs.29.77[25.75,40.97]s,Z=-4.832).The difference of the above between two group was statistically significant(all P＜0.01).(3)Brain parenchymal fraction in CSVD group was lower than control group,and there was significant difference between two group([78.2±4.3]％vs.[80.9±3.7]％,t=2.079,P＜0.05).VBM analysis showed that gray matter volume of right inferior temporal gyrus(rITG)and right Crus 2 of cerebellar hemisphere(rCERCRU2)in CSVD group was significantly lower than control group(both P＜0.05 and corrected by false discovery rate).(4)Partial correlation analysis showed a positive correlation between gray matter volume in rITG and AVLT-short delay recall score(r=0.543,P=0.036).Conclusions CSVD patients with cognitive impairment had gray matter atrophy in rITG and rCERCRU2 and the gray matter volume in rITG was correlated with delayed memory impairment.The results of this study need to be further verified.

Objective:The clinical characteristics and prognosis of 20 patients with small B-lymphocyte proliferative disease with t (14;19) (q32; q13) were analyzed to improve the understanding of such rare cases.Methods:The clinical data of 20 patients with t (14; 19) (q32; q13) small B lymphocyte proliferative disease treated in the First Affiliated Hospital of Nanjing Medical University from April 2013 to December 2020 were retrospectively collected and analyzed. Among them, 10 cases were chronic lymphocytic leukemia (CLL) and 10 cases were other small B-cell malignancies.Results:Among the 20 cases, 10 were male and 10 were female, and the median age at diagnosis was 53.5 (35-88) years old. All patients had absolute lymphocytosis, 19 patients had lymphadenopathy, and 10 patients had splenomegaly. With a median follow-up of 36 (4-163) months, three patients died, and 11 patients had a time to treatment (TTT) ≤12 months. Ten patients (50%) were accompanied by +12, two patients (2/17, 12%) were accompanied by 13q-. Moreover, we found that t (14;19) was associated with unmutated immunoglobulin heavy-chain variable (IGHV) somatic mutation (17/19, 89%) and a biased use of IGHV4-39 (7/17, 41%) was observed. Next-generation sequencing detected one or more gene mutations in 14 (14/17, 82%) cases and a total of 25 gene mutations had been revealed, of which the most frequent were NOTCH1 (35%) , followed by SF3B1 (24%) and KMT2D (18%) . For 10 CLL patients, five (50%) were defined as Rai Ⅲ/Binet C. It is noteworthy that among the 20 cases, two cases actually involved Richter transformation.Conclusions:Small B-cell malignant tumors with abnormal t (14; 19) show unique clinical biological characteristics, often accompanied by a variety of adverse prognostic factors, and tend to have an aggressive clinical course.

OBJECTIVE：To compare the chemical constituents of petroleum ether fraction from ethanol extract of Aconitum sinomontanum before and after processing. METHODS ：After A. sinomontanum was purified with water ，the raw product decoction pieces were prepared ；the raw decoction pieces were steamed with licorice juice under high pressure to prepare processed decoction pieces of A. sinomontanum . The petroleum ether fractions of raw product and processed product were obtained after ultrasonic extraction with 95％ ethanol. The chemical constituents in the samples were analyzed by GC-MS. NIST 2014 mass spectrometry database was used to compare and match the components . The peak area normalization method was used to determine the relative percentage content of each component. RESULTS ：Before and after processing ，fatty acids and esters were the main components in the petroleum ether fraction from ethanol extract. Totally 18 chromatographic peaks were detected in the detection pieces of raw product，and 13 compounds were identified ，accounting for 94.60％ of the total content of volatile components. The components with relatively high content were （Z，Z，Z）-9，12，15-octadecatrienoic acid （26.13％），hexadecanoic acid ethyl ester （25.27％）， palmitoleic acid （10.84％），ethyl linoleic acid （10.67％），（Z，Z）-9，12-octadecenoic acid methyl ester （6.66％），pentadecanoic acid（5.11％）and so on. Totally 25 chromatographic peaks were detected in the decoction pieces of processed products，and 18 components were identified ，accounting for 82.40％ of the total content of volatile components. The components with relatively high content were palmitoleic acid （18.95％），（Z，Z）-9，12-octadecenoic acid methyl ester （17.93％），hexadecanoic acid ethyl ester（11.94％），（Z，Z，Z）-9，12，15-octadecatrienoic acid （10.54％），（Z，Z）-9，12-octadecenoic acid （5.51％），（Z）-11-hexadecanoic acid（5.30％）and so on. After processing ，7 new components were added ，5 of which were identified as （－）-eucalyptus globulus alcohol，ethyl 2-methyltetrade-canoate，6-methyl-4-phenylcoumarin，β-sitosterol，heptadecane. After processing ，no components disappeared，and the content of some components increased or decreased. CONCLUSIONS ：After processing ，the volatile components in the petroleum ether fraction from ethanol extract of A. sinomontanum are different ，and（－）-eucalyptus globulus alcohol and other components are added after processing.

Objective:The study aimed to analyze the clinical features and prognosis of chronic lymphocytic leukemia (CLL) with t (14;18) (q32;q21) and conduct a literature review.Methods:The clinical data of 8 patients with CLL carrying t (14;18) (q32;q21) seen in Jiangsu Province Hospital from November 2009 to November 2019 were collected and analyzed.Results:Among the 8 cases, 7 were male and 1 was female. The median age at diagnosis was 70 years old. The immunophenotype score was 5 in 3 patients. 4 patients were scored 4 and the remaining one scored 3. The bone marrow histopathology showed the typical manifestation of CLL. Karyotype analysis showed that all the cases carried t (14;18) (q32;q21) in the stemline. The t (14;18) (q32;q21) showed as the sole abnormality in 3 cases, with +12 in 4, and with 13q- in 1 case. 13q- was found in another 3 patients by FISH. Immunoglobulin heavy chain gene (IGHV) mutation status was detected in 6 cases and all of them were mutated. None of them used IGHV3-21. Only 1 case harbored TP53 mutation and no TP53, SF3B1, NOTCH1, or MYD88 mutations were found in the remaining cases who underwent the relevant tests. At a median follow-up of 30.9 months, 1 case died. The remaining 7 cases survived and 3 of them have not reached the treatment indication. 4 patients who received chemotherapy or immunotherapy were stable.Conclusions:The t (14;18) (q32;q21) is rare in CLL and often accompanied by +12 and mutated IGHV. CLL with t (14; 18) (q32; q21) tends to have a good prognosis.

Sex reversal, representing extraordinary sexual plasticity during the life cycle, not only triggers reproduction in animals but also affects reproductive and endocrine system-related diseases and cancers in humans. Sex reversal has been broadly reported in animals; however, an integrated resource hub of sex reversal information is still lacking. Here, we constructed a comprehensive database named ASER (Animal Sex Reversal) by integrating sex reversal-related data of 18 species from teleostei to mammalia. We systematically collected 40,018 published papers and mined the sex reversal-associated genes (SRGs), including their regulatory networks, from 1611 core papers. We annotated homologous genes and computed conservation scores for whole genomes across the 18 species. Furthermore, we collected available RNA-seq datasets and investigated the expression dynamics of SRGs during sex reversal or sex determination processes. In addition, we manually annotated 550 in situ hybridization (ISH), fluorescence in situ hybridization (FISH), and im-munohistochemistry (IHC) images of SRGs from the literature and described their spatial expression in the gonads. Collectively, ASER provides a unique and integrated resource for researchers to query and reuse organized data to explore the mechanisms and applications of SRGs in animal breeding and human health. The ASER database is publicly available at http://aser.ihb.ac.cn/.