OBJECTIVES: To explore the association of the expression of WW domain-containing ubiquitin E3 ligase 1 (WWP1) with immune infiltration in tumor microenvironment (TME) of ovarian cancer. METHODS: Ovarian cancer patient data from The Cancer Genome Atlas (TCGA) were used to analyze the association of WWP1 expression with patient prognosis. TISCH2 was utilized to analyze the changes in immune cell subtypes in TME of metastatic tumor and after chemotherapy. The impact of WWP1 on immune cell infiltration, somatic copy number alterations of WWP1 and evolution of immune cell subtypes was evaluated using TIMER and TIGER pseudo-time analysis. A deep learning model was used to analyze TCGA pathological images to investigate the effect of WWP1 on TME of ovarian cancer. RNA-seq analysis was conducted to identify the differentially expressed genes in WWP1-overexpressing SKOV3 cells and validate immune infiltration. Multicolor immunofluorescence assay was used to analyze the immune markers in SKOV3 and SKOV3/DDP cell xenografts in nude mice. RESULTS: The patients with high WWP1 expression levels had significantly lower overall survival rate (P=0.0012). High WWP1 expression levels and Stage IV disease were both associated with a poor prognosis (P<0.05). In metastatic ovarian cancer or after chemotherapy, the percentages of malignant tumor cells and tumor-associated fibroblasts increased in the TME, accompanied by elevated WWP1 levels. WWP1 expression level was positively correlated with pro-tumorigenic immunosuppressive cells (r=0.1323-0.3955, P<0.05) and negatively with tumor-inhibiting immune cells (r=-0.1949- -0.1333, P<0.05). Specific copy number alterations of WWP1 also influenced CD8⁺ T cell percentage and neutrophil infiltration levels in the TME. RNA-seq analysis of WWP1-overexpressing SKOV3 cells and immunofluorescence assay of the tumor-bearing mice yielded findings consistent with those of bioinformatics analysis. CONCLUSIONS WWP1 may serve as a prognostic biomarker and a potential target for immune regulation in the TME of ovarian cancer.
Female ; Ovarian Neoplasms/genetics* ; Humans ; Ubiquitin-Protein Ligases/metabolism* ; Tumor Microenvironment/immunology* ; Animals ; Mice ; Cell Line, Tumor ; Mice, Nude ; Prognosis ; Gene Expression Regulation, Neoplastic

Objective To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods,and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques.This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix.Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique.The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases.Targets of immune-related disease were obtained through DisGeNET,OMIM,TTD,and MalaCards databases.The network of"components absorbed into blood from Hedysari Radix-immune-related diseases"was then constructed.GO and KEGG enrichment analysis and mapped the PPI network were performed.Molecular docking and molecular dynamics techniques were applied for validation.Results A total of 8 prototype components absorbed into blood,synergistically acting on 101 targets,were identified by UPLC-MS/MS.They mediated 538 biological processes including immune response,positive regulation of gene expression,receptor binding,and cytokine activity.Meanuhile,116 signaling pathways,such as HIF-1,Toll-like receptor,JAK-STAT,T cell receptor,PI3K-Akt,and FoxO etc.were involved.The core targets were MAPK14,PTGS2,MMP9,PPARG,CCND1,etc..The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14.And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity.Conclusion The results of serum pharmacochemistry,network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity.The aim of this study is to provide scientific basis for its immunomodulatory mechanism.

DNMT3A encodes a DNA methyltransferase involved in development, cell differentiation, and gene transcription, which is mutated and aberrant-expressed in cancers. Here, we revealed that loss of DNMT3A promotes malignant phenotypes in lung cancer. Based on the epigenetic inhibitor library synthetic lethal screening, we found that small-molecule HDAC6 inhibitors selectively killed DNMT3A-defective NSCLC cells. Knockdown of HDAC6 by siRNAs reduced cell growth and induced apoptosis in DNMT3A-defective NSCLC cells. However, sensitive cells became resistant when DNMT3A was rescued. Furthermore, the selectivity to HDAC6 inhibition was recapitulated in mice, where an HDAC6 inhibitor retarded tumor growth established from DNMT3A-defective but not DNMT3A parental NSCLC cells. Mechanistically, DNMT3A loss resulted in the upregulation of HDAC6 through decreasing its promoter CpG methylation and enhancing transcription factor RUNX1 binding. Notably, our results indicated that HIF-1 pathway was activated in DNMT3A-defective cells whereas inactivated by HDAC6 inhibition. Knockout of HIF-1 contributed to the elimination of synthetic lethality between DNMT3A and HDAC6. Interestingly, HIF-1 pathway inhibitors could mimic the selective efficacy of HDAC6 inhibition in DNMT3A-defective cells. These results demonstrated HDAC6 as a HIF-1-dependent vulnerability of DNMT3A-defective cancers. Together, our findings identify HDAC6 as a potential HIF-1-dependent therapeutic target for the treatment of DNMT3A-defective cancers like NSCLC.

Malignant tumors are one of the main causes of human death worldwide and pose a serious threat to human health. The current treatment methods are mainly the combination of chemotherapeutics， surgery， radiotherapy， or hormone therapy. The treatment process has limitations such as multidrug resistance， non-selective targeting of cancer cells， and drug toxicity. With the development and application of traditional Chinese medicine （TCM）， Chinese medicine has the characteristics of multi-angle and multi-mechanism coordination and slight toxic and side effects. It can effectively inhibit tumor proliferation， differentiation， and metastasis， and avoid drug resistance， serving as the focus of current tumor treatment research. Hedysari Radix， one of the genuine medicinal materials in Gansu province， is a tonic Chinese medicine with a wide range of pharmacological effects such as anti-inflammation， immune regulation， anti-oxidation， prevention and treatment of diabetic complications. In the majority of the ancient works on herbs of the past dynasties， Hedysari Radix was included under the item of Astragali Radix and used as Astragali Radix. Hedysari Radix is superior to Astragali Radix in enhancing immunity， scavenging free radicals， and resisting liver fibrosis. Studies have found that the effective components of Hedysari Radix have a prominent anti-tumor effect and a significant inhibitory effect on various malignant tumors such as liver cancer， bladder cancer， gastric cancer， breast cancer， and colorectal cancer. They can also combine with clinical anti-cancer drugs to reduce the toxic and side effects of chemotherapy drugs and improve the tolerance of patients during chemotherapy. On the basis of current research， this study summarized the mechanism of Hedysari Radix active components in inducing cell apoptosis， blocking cell cycle， inhibiting tumor cell proliferation， migration， and invasion， regulating micro mRNA （miRNA）， inducing cell autophagy， enhancing immune regulation， as well as reducing toxicity and enhancing efficiency and sensitization with clinical chemotherapeutics， and systematically explained the anti-tumor mechanism of Hedysari Radix active components， aiming to provide a basic reference for the further exploration of the anti-tumor mechanism of Hedysari Radix and the further development and utilization of its effective components.

Qi-Yu-San-Long decoction(QYSLD)is a traditional Chinese medicine that has been clinically used in the treatment of non-small-cell lung cancer(NSCLC)for more than 20 years.However,to date,metabolic-related studies on QYSLD have not been performed.In this study,a post-targeted screening strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight full infor-mation tandem mass spectrometry(UPLC-QTOF-MSE)was developed to identify QYSLD-related xeno-biotics in rat urine.The chemical compound database of QYSLD constituents was established from previous research,and metabolites related to these compounds were predicted in combination with their possible metabolic pathways.The metabolites were identified by extracted ion chromatograms using predicted m/z values as well as retention time,excimer ions,and fragmentation behavior.Overall,85 QYSLD-related xenobiotics(20 prototype compounds and 65 metabolites)were characterized from rat urine.The main metabolic reactions and elimination features of QYSLD included oxidation,reduction,decarboxylation,hydrolysis,demethylation,glucuronidation,sulfation,methylation,deglycosylation,acetylation,and associated combination reactions.Of the identified molecules,14 prototype compounds and 58 metabolites were slowly eliminated,thus accumulating in vivo over an extended period,while five prototypes and two metabolites were present in vivo for a short duration.Furthermore,one pro-totype and five metabolites underwent the process of"appearing-disappearing-reappearing"in vivo.Overall,the metabolic profile and characteristics of QYSLD in rat urine were determined,which is useful in elucidating the active components of the decoction in vivo,thus providing the basis for studying its mechanism of action.

Hepatocellular carcinoma (HCC) is the most common pathological type of primary liver cancer, with high fatality rate. The pathogenesis of HCC is complex, and the specific occurrence and development mechanism is still in the exploratory stage. CircRNA is a special endogenous noncoding RNA and mainly participates in the regulation of gene expression at the transcriptional and posttranscriptional levels. By regulating gene transcription, it acts as a molecular sponge of miRNA, participates in protein translation, and interacts with RNA binding protein (RBP). CircRNA is involved in the occurrence and development of HCC. Its abnormal expression in HCC cells is related to the pathological characteristics of HCC tissue, regulates the expression of downstream target genes, miRNA and proteins, participates in the proliferation, migration, invasion and apoptosis of HCC cells, and regulates tumor microenvironment and signal pathways, suggesting that circRNA may be a potential novel biomarker and therapeutic target for the diagnosis and prognosis of HCC. This paper reviews the biological mechanism of circRNA, its role in HCC, and research progress in diagnosis and treatment.

Objective:To evaluate the prognostic value of amplitude-integrated electroencephalogram(aEEG) on predicting long-term neurodevelopmental outcome of preterm infants.Methods:Literatures were searched in PubMed, the Cochrane library, Web of science, Wanfang database, CNKI and CBM database from inception to August, 2020.The studies which investigated the prognostic value of aEEG on neurodevelopmental outcome of preterm infants were included.Articles screening, data extraction and quality assessment were accomplished by two investigators independently, and statistical analyses were performed by Meta-disc1.4.Results:Six studies were included with 557 cases.The Meta-analysis revealed that the pooled sensitivity was 0.81 (95% CI 0.75-0.87), specificity was 0.68 (95% CI 0.63-0.73), positive likelihood ratio was 2.32(95% CI 1.52-3.52), negative likelihood ratio was 0.27(95% CI 0.19-0.37)and the area under the curve of summary receiver operating characteristic curve was 0.79(95% CI 0.75-0.82). Burst-suppression, continuous low voltage and flat trace were regard as a index of poor neurodevelopmental prognosis. Conclusion:aEEG is a valuable tool for predicting the outcome of long-term neurodevelopment in preterm infants.

Objective:To compare the efficacy of haplotype hematopoietic stem cell transplantation (HIDT) and sibling matched hematopoietic stem cell transplantation (MSDT) in the treatment of complete remission (CR) acute T-lymphoblastic leukemia (T-ALL) .Methods:We retrospectively analyzed the clinical characteristics and outcomes of 98 patients who underwent HSCT in Hebei Yanda Ludaopei hospital with HID ( n=81) or ISD ( n=17) between May 2012 and May 2016. Results:The incidence of grades 2-4 and 3-4 acute-versus-host disease 100 days after HSCT were 51.9% (95% Confidence interval [ CI] 42.0%-64.0%) vs 29.4% (95% CI 14.1%-61.4%) ( P=0.072) and 9.8% (95% CI 5.1%-19.1%) vs 11.8% (95% CI 3.2%-43.3%) ( P=1.000) for HIDT and MSDT. The 100-day cumulative incidences of CMV and EBV viremia were 53.1% (95% CI 43.3%-65.2%) vs 29.4% (95% CI 14.1%-61.4%) ( P=0.115) and 35.8% (95% CI 26.8%-47.9%) vs11.8% (95% CI 3.2%-43.3%) ( P=0.048) . The 5-year overall survival, leukemia-free survival, cumulative incidences of relapse, and no-relapse mortality were 60.5% (95% CI 5.4%-49.0%) vs 68.8% (95% CI 11.8%-40.0%) ( P=0.315) , 58.0% (95% CI 5.5%-46.5%) vs 68.8% (95% CI 11.8%-40.0%) ( P=0.258) , 16.1% (95% CI 9.8%-26.4%) vs 11.8% (95% CI 3.2%-43.3%) ( P=0.643) , 25.9% (95% CI 17.9%-37.5%) vs 19.4% (95% CI 6.9%-54.4%) ( P=0.386) for HIDT and MSDT, respectively. Conclusion:HID could be a valid alternative donor for patients with T-ALL in CR lacking an identical donor.

Objective:To study the clinical efficacy of chimeric antigen receptor T-cell (CART) treatment followed by a second allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with B-cell acute lymphoblastic leukemia (ALL) who relapsed following the first HSCT.Methods:Retrospective analysis of the clinical characteristics and prognosis of 41 patients with B-cell ALL who received a second allo-HSCT from October 2015 to June 2020 in Hebei Yanda Lu Daopei Hospital. After the first HSCT, all patients received CD19-CART, or CD22-CART treatment following a relapse of bone marrow morphology or extramedullary leukemia.Results:A total of 41 patients (male, 21; female, 20) were included in this study. The median age at the second HSCT was 16 (3-46) years. There were 31 cases of bone marrow recurrence (75.6%) , 5 cases of extramedullary recurrence (12.2%) , and 5 cases of bone marrow and extramedullary recurrences (12.2%) . After relapse, 35 patients (85.4%) received CD19-CART treatment, 2 patients received CD22-CART treatment (4.9%) , and 4 patients received CD19-CART and CD22-CART treatments (9.8%) . The expected 3-year overall survival (OS) , leukemia-free survival, cumulative relapse incidence, and non-relapse mortality (NRM) of patients after the second HSCT were 48.9% (95% CI 23.0%-70.6%) , 41.8% (95% CI 17.3%-64.9%) , 8.8% (95% CI 2.9%-26.4%) , and 51.1% (95% CI 31.2%-83.6%) , respectively. The 1-year OS of patients who relapsed ≤6 months and >6 months after the first HSCT were 45.0% (95% CI 12.7%-73.5%) and 75.0% (95% CI 51.4% -88.8%) ( P=0.017) , respectively. Conclusion:CART bridging in the second HSCT enables some B-cell ALL patients who relapsed after the first HSCT to achieve long-term survival. However, because of the high NRM, further modifications could help improve the outcome.

Objective:To explore the risk factors influencing the short-term mortality of patients with sepsis in intensive care unit (ICU) and the combined value of predicting prognosis.Methods:A retrospective analysis was performed on 104 patients with sepsis admitted to emergency ICU of Jiangsu Provincial Hospital of Traditional Chinese Medicine from January 2018 to August 2019. Multiple general information containing gender, age, past history as well as complications and sequential organ failure assessment (SOFA) score, mean arterial pressure (MAP), blood routine examination, hepatic and renal function, coagulation indicators and procalcitonin (PCT) were collected within 24 hours of admission. Patients were divided into death group and survival group according to the 28-day outcome. Univariate and multivariate Logistic regression analysis were used to find the effective factors influencing the prognosis of sepsis. Receiver operating characteristic (ROC) curve was drawn to evaluate the value of related indexes in predicting the prognosis of sepsis. Correlation between parameters that might be relevant to disease severity and SOFA score was evaluated by Pearson or Spearman correlation analysis.Results:104 patients were enrolled for final analysis, of whom 60 patients survived, while the others died with a 28-day mortality of 42.3%. ① Univariate analysis results: the incidence of acute kidney injury (AKI), SOFA score, serum creatinine (SCr), D-dimer, activated partial thromboplastin time (APTT), international normalized ratio (INR) and PCT in the death group were significantly higher than those in the survival group [incidence of AKI: 70.5% (31/44) vs. 36.7% (22/60), SOFA score: 11.0 (8.0, 13.0) vs. 8.0 (6.2, 10.0), SCr (μmol/L): 108.8 (65.5, 235.6) vs. 75.1 (55.1, 109.5), D-dimer (mg/L): 4.1 (1.6, 11.6) vs. 2.1 (1.2, 4.3), APTT (s): 42.6 (37.7, 55.7) vs. 40.3 (35.9, 44.7), INR: 1.3 (1.2, 1.5) vs. 1.2 (1.1, 1.4), PCT (μg/L): 3.1 (0.4, 39.9) vs. 0.3 (0.1, 3.4), all P < 0.05]. ② Multivariate Logistic regression analysis results: all indicators of univariate analysis were included in the multivariate Logistic regression model considering interaction between each variable. Multivariate Logistic regression analysis was repeated based on conditional backward method. Age, SOFA score, MAP, neutrophil (NEU), lymphocyte (LYM) and APTT were automatically selected by SPSS software to build the predicting model. Analysis results showed that SOFA score, NEU and LYM were independent risk factors for the short-term prognosis of sepsis [SOFA score: odds ratio ( OR) = 1.22, 95% confidence interval (95% CI) was 1.04-1.44, P = 0.02; NEU: OR = 1.14, 95% CI was 1.03-1.26, P = 0.01; LYM: OR = 0.79, 95% CI was 0.66-0.95, P = 0.01]. ③ ROC curve analysis results: the above six-variable prediction model had the optimal fitting degree defaulted by SPSS. ROC curve showed that the combination of age [area under ROC curve (AUC) = 0.60], SOFA score (AUC = 0.71), MAP (AUC = 0.53), NEU (AUC = 0.59), LYM (AUC = 0.54) and APTT (AUC = 0.61) had better sensitivity (79.5%) and specificity (65.0%) as well as the maximal AUC (AUC = 0.75), which suggested that combined prediction had higher diagnostic value in predicting the short-term prognosis of sepsis.④ Correlation analysis showed that NEU, D-dimer, prothrombin time (PT), APTT, INR and PCT were positively correlated with SOFA score ( r values were 0.26, 0.28, 0.21, 0.22, 0.10, 0.38, respectively, all P < 0.05). Conclusions:SOFA score, NEU and LYM were independent risk factors for the short-term prognosis of sepsis. The combination of age, SOFA score, MAP, NEU, LYM and APTT were more accurate than any single factor in predicting the short-term prognosis of sepsis and had higher diagnostic value. NEU, D-dimer, PT, APTT, INR and PCT were correlated with SOFA score.