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.

Head and neck squamous cell carcinoma (HNSCC) is a tumor characterized by immunosuppressive tumor microenvironment (TME) and poor prognosis. Its complex immune evasion mechanisms are primarily related to T cell dysfunction and the suppression of anti-tumor immune responses. Immunotherapy aims to modulate the patient’s immune system to recognize and eliminate tumor cells, thereby achieving therapeutic goals. Studies have demonstrated that the TME plays a pivotal role in HNSCC pathogenesis, facilitating tumorigenesis, progression, and therapy resistance, ultimately contributing to adverse clinical outcomes. Advances in technology have deepened understanding of the TME, paving the way for novel therapeutic interventions in HNSCC. This review comprehensively summarizes the efficacy and safety of TME-targeted immunotherapies, integrating evidence from published clinical trials, while proposing insights for future research to develop more effective therapeutic strategies.

OBJECTIVE To identify the key genes associated with lymph node metastasis in head and neck squamous carcinoma(HNSCC)and construct a prognostic model based on The Cancer Genome Atlas(TCGA)database.METHODS Differentially expressed genes(DEGs)between tumor tissues and normal tissues in the HNSCC dataset in the TCGA database were screened by R software,and gene modules related to lymph node metastasis were screened by weighted gene co-expression network(weighted gene co-expression network analysis,WGCNA).Prognostic risk models were constructed by univariate cox regression and Lasso regression analyses.Survival analyses and ROC curves were performed to verify the Reliability of prognostic models.CIBERSORT,TIMER and ESTIMATE algorithms analysed the differences in the tumor micro environment(TME)of different risk groups.RESULTS There were 2 565 DEGs screened,and a set of gene modules highly correlated with disease prognosis and lymph node metastasis were obtained by WGCNA analysis,and correlation analysis verified that the expression of genes in this gene module was highly correlated with lymph node metastasis.Univariate cox regression and Lasso regression were used to identify 6 key prognostic genes:CDKN2A,CCNE2,KNSTRN,AURKA,KPNA2,and ORC1.A prognostic model was constructed based on the 6 genes,and survival analysis showed that the prognosis of the high-risk group was significantly worse than that of the low-risk group(P<0.0001).The ROC curves demonstrated the good predictive performance of this prognostic model.CIBERSORT analyses revealed differences in the immune microenvironment of tumors in different risk groups.CONCLUSION The 6 key prognostic genes screened were helpful in predicting the prognosis of HNSCC patients and were closely associated with the immune microenvironment of HNSCC,suggesting that they may serve as potential therapeutic targets.

Objective To observe MRI features of H3K27M mutant type and wild type astrocyte differentiated diffuse midline glioma(DMG)in spinal cord.Methods Totally 91 patients with astrocyte differentiation diffuse midline glioma(DMG)in spinal cord confirmed by pathology were retrospectively enrolled and divided into mutant group(n=44)and wild group(n=47)according to H3K27M status.Clinical and MRI manifestations were compared between groups,and logistic regression analysis was used to screen the impact factors of H3K27M mutation.Results The incidence of peritumoral edema and spinal cord cavity in mutant group were lower than those in wild group(both P＜0.05),while no significant difference of other parameters was found between groups(all P＞0.05).All clinical and MRI parameters were included in logistic regression analysis,and the result showed that they were not influencing factors of H3K27M mutation(all P＞0.05).Conclusion The incidence of peritumoral edema and spinal cord cavity in spinal cord H3K27M mutant type astrocyte differentiated DMG were lower than those of wild type,yet not sufficient to be regarded as impact factors for predicting H3K27M mutation of DMG.

Objective:To analyze the factors influencing prognosis of intrahepatic cholangiocarcinoma (ICC) after surgical resection.Methods:The clinical data of patients diagnosed with ICC and who underwent surgical resection from December 2015 to December 2019 at the Fifth Medical Center of PLA General Hospital were retrospectively analyzed. Of 39 patients who were included in this study, there were 23 males and 16 females, with age of (54.1±7.2) years old. The body mass index, hepatitis B virus infection status, tumor diameter, degree of differentiation, microvascular tumor thrombus, lymph node metastasis, and serum levels of carbohydrate antigen 19-9 (CA19-9) were analyzed as risk factors affecting postoperative recurrence and survival.Results:The median times to recurrence were significantly better in patients with a tumour length <5 cm (11 vs. 5 months), patients without microvascular tumor thrombus (54 vs. 6 months) and patients without lymph node metastasis (8 vs. 5 months) (all P<0.05). The median survival of patients with CA19-9≥100 U/ml was significantly shorter than that of patients with CA19-9<100 U/ml, (9 vs. 27 months, P<0.05). Tumor diameter>5 cm, microvascular tumor thrombus, lymph node metastasis, and CA19-9 ≥100 U/ml are risk factors affecting the recurrence time after ICC resection, CA19-9 ≥100 U/ml is a risk factor affecting survival time after ICC resection. Conclusion:Tumor diameter, microvascular tumor thrombus, lymph node metastasis and CA19-9 can be used to estimate the risk of ICC recurrence, and CA19-9 level can be used to estimate postoperative survival of ICC patients after resection.

Objective To explore the effect of mechanical stimulation on polarity of macrophages. Methods RAW264.7 cells were stimulated with tensile stretch at various amplitude and time, then cell viability was assessed with cell count kit-8 (CCK-8) for determining the stimulation parameters. RAW264.7 cells were induced to M1 type, then tensile stretch at 10% amplitude and 2 Hz was applied to M1 cells. CCK-8 and flow cytometry were used to detect the effects of tensile stretch on cell activity and apoptosis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the effect of tensile stretch on M1 type macrophage related gene expression. Results After stimulation for 3 hours, tensile stretch at 15% or 20% amplitude and 2 Hz significantly inhibited cell viability (P<0.05), while tensile stretch at 10% amplitude and 2 Hz did not inhibit the viability of RAW264.7 cells (P>0.05). Tensile stretch at 10% amplitude and 2 Hz neither inhibited viability nor cause apoptosis of M1 type macrophages. The expression of inflammation-related genes including interleukin-1β(IL-1β) and tumor necrosis factor-α (TNF-α) of M1 type macrophages was significantly down-regulated with tensile stretch at 10% amplitude and 2 Hz (P＜0.05). Conclusions Mechanical stimulation at 10% amplitude and 2 Hz can inhibit M1 type macrophages and promote the polarization from M1 to M2. Mechanical stimulation may become a method for treating inflammation-related diseases.

Objective :To explore therapeutic effect of emergency percutaneous coronary intervention (PCI) combined ticagrelor on patients with acute ST elevation myocardial infarction (STEMI ) and its influence on level of tumor necrosis factor (TNF)—α.Methods : A total of 98 patients with acute STEMI treated in our hospital from Jan 2016 to Jan 2017 were selected .Patients were randomly and equally divided into clopidogrel group and ticagrelor group , each group received corresponding medication before PCI .LVEF ,LVEDd ,TNF—α level before and one month after treatment and TIMI grade before and after PCI ,recanalization time ,postoperative corrected TIMI frame (CTFC) , percentages of thrombus aspiration ,auxiliary IABP and no—reflow ,and incidence of adverse events during six—month follow—up after PCI were observed and compared between two groups .Results : Compared with before PCI and treatment ,there were significant rise in LVEF on one month after treatment and TIMI grade after PCI ,and signifi—cant reductions in LVEDd and TNF—α level in two groups on one month after treatment , P＝0.001 all.Compared with clopidogrel group on one month after treatment ,there was significant rise in LVEF [ (49.80 ± 4.17 )% vs. (57.32 ± 5.10)%] ,and significant reductions in LVEDd [ (57. 94 ± 4. 70) mm vs .(47.11 ± 3.49) mm] ,TNF—α level [ (17. 82 ± 2.84) pg／ml vs .(8.40 ± 2. 04 ) pg／ml] and postoperative percentage of no—reflow (24.44% vs. 4.44%) in ticagrelor group , P＜0. 01 all ,there were no significant difference in other operative related indexes be—tween two groups , P＞0.05 all.After six—month follow—up ,incidence rate of composite endpoint events in ticagrelor group was significantly lower than that of clopidogrel group (11. 11% vs.31. 11%) , P＝0.021 ,there were no sig—nificant difference in percentages of in—stent thrombus ,infarct related artery revascularization and recurrent unsta—ble angina pectoris between two groups , P＞0.05 all .Conclusion : Short—term therapeutic effect of emergency PCI combined ticagrelor is significant .It can significantly improve prognosis ,which is worth extending .

Uncovering the functionally essential variations related to tumorigenesis and tumor pro-gression from cancer genomics data is still challenging due to the genetic diversity among patients, and extensive inter-and intra-tumoral heterogeneity at different levels of gene expression regulation, including but not limited to the genomic, epigenomic, and transcriptional levels. To minimize the impact of germline genetic heterogeneities, in this study, we establish multiple primary cultures from the primary and recurrent tumors of a single patient with hepatocellular carcinoma (HCC). Multi-omics sequencing was performed for these cultures that encompass the diversity of tumor cells from the same patient. Variations in the genome sequence, epigenetic modification, and gene expression are used to infer the phylogenetic relationships of these cell cultures. We find the discrepancy among the relationships revealed by single nucleotide variations (SNVs) and transcriptional/epigenomic pro-files from the cell cultures. We fail to find overlap between sample-specific mutated genes and differ-entially expressed genes (DEGs), suggesting that most of the heterogeneous SNVs among tumor stages or lineages of the patient are functionally insignificant. Moreover, copy number alterations (CNAs) and DNA methylation variation within gene bodies, rather than promoters, are significantly correlated with gene expression variability among these cell cultures. Pathway analysis of CNA/DNA methylation-related genes indicates that a single cell clone from the recurrent tumor exhibits distinct cellular characteristics and tumorigenicity, and such an observation is further confirmed by cellular experiments both in vitro and in vivo. Our systematic analysis reveals that CNAs and epigenomic changes, rather than SNVs, are more likely to contribute to the phenotypic diversity among subpop-ulations in the tumor. These findings suggest that new therapeutic strategies targeting gene dosage and epigenetic modification should be considered in personalized cancer medicine. This culture model may be applied to the further identification of plausible determinants of cancer metastasis and relapse.

Objective The aims of the study were to investigate the effects of human islet amyloid polypeptide (hIAPP) on autophagy in INS-1 cells and its underlying mechanism,and to explore the role of autophagy in hIAPP-induced cytotoxicity and oxidative stress.Methods INS-1 cells were treated with hIAPP (10 μmol/L) for 24 h in the presence or absence of N-acetyl-L-cysteine (NAC),compound C,5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) and 3-methyladenine (3-MA),respectively.Transmission electron microscopy was used to observe the number of autophagosome in cells.Cell viability was determined by methyl thiazolyl tetrazolium (MTT) test.2',7'-dichlorofluorescin diacetate (DCFH-DA) assay was used to measure the relative levels of reactive oxygen species (ROS).Western blot was used to detect expression of adenosine monophosphate-activated protein kinase (AMPK) and autophagic markers p62 and microtubule associated protein 1 light chain3 (LC3).Results Treatment of INS-1 cells with hIAPP resulted in a significant increase in the number of autophagosomes and the expression of LC3-Ⅱ/LC3-Ⅰ (both P＜0.05).Meanwhile,treatment of INS-1 cells with hIAPP enhanced the level of ROS to 1.76 times of control cells (P＜0.01).Co-treatment with NAC,an antioxidant,inhibited hIAPP-induced ROS generation,and the expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK in the INS-1 cells (all P＜0.05).Pretreatment of INS-1 cells with AMPK inhibitor compound C suppressed hIAPP and AICAR,an activator of AMPK,induced expression of LC3-Ⅱ/LC3-Ⅰ and p-AMPK (all P＜0.05).Autophagic inhibitor 3-MA and compound C aggravated the hIAPP-induced cell death and ROS generation in INS-1 cells (All P＜0.05).The cytotoxic effects of hIAPP were significantly attenuated by co-treatment with AICAR (P＜0.05).Conclusion Autophagy may act as an adaptive mechanism to alleviate hIAPP-induced oxidative damage and toxicity in INS-1 cells.

Nicotinamide phosphoribosyltransferase( NAMPT) is the rate-limiting enzyme that catalyzes the first step in Nicotinamide adenine dinucleotide( NAD) biosynthesis and regulates biological processes such as metab-olism,growth and apoptosis of cells.In the past few years,a large number of studies have found that expression level of NAMPT is increased in a variety of malignancies,which is associated with the progression and treatment of malig-nant tumors.The research of the mechanism of NAMPT in malignant tumors and molecule inhibitors of the NAMPT have brought new hope and prospect for the treatment of malignant tumors.This article makes a brief review of NAMPT research status in malignant tumors.