Objective: To investigate the protective effects of morusin on lipopolysaccharide (LPS)-induced acute liver injury in mice and its underlying mechanisms. Methods: Thirty-two male specific pathogen-free (SPF) C57BL/6J mice were randomly divided into four groups (n = 8 per group): control, LPS, low-dose morusin (morusin-L, 10 mg/kg), and high-dose morusin (morusin-H, 20 mg/kg) groups. The mice in each group were administered the corresponding drugs or normal saline via continuous gavage daily for 16 consecutive days. Except for control group, which received an equal volume of normal saline, other groups were intraperitoneally injected with LPS (5 mg/kg) 2 h after the last gavage to establish the acute liver injury model. Serum and liver tissues were collected for subsequent analysis 6 h after LPS injection. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected with biochemical methods. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in serum were measured by enzyme-linked immunosorbent assay (ELISA). Hepatic pathological changes were evaluated by hematoxylin-eosin (HE) staining. The 16S ribosomal RNA (16S rRNA) sequencing was performed to assess the composition of intestinal flora, linear discriminant analysis effect size (LEfSe) was applied for multi-level species discrimination, and Spearman’s correlation analysis was performed. The liver tissues of mice with acute liver injury were analyzed by RNA sequencing (RNA-seq) technology to identify differentially expressed genes (DEGs), and then enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was conducted. The expression levels of selected genes was validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR), while immunohistochemistry (IHC) was performed to examine the expression levels of IL-6, myeloid differentiation primary response 88 (MYD88), and toll-like receptor 2 (TLR2). Results: Morusin significantly reduced the serum levels of ALT, AST, and inflammatory factors (TNF-α, IL-6, and IL-1β) (P < 0.05, P < 0.01, or P < 0.001), while alleviating the hepatic pathological damage in mice. Based on efficacy comparisons, morusin-H group was selected for subsequent microbiome and transcriptome analyses. Microbiome analysis revealed that morusin-H effectively mitigated LPS-induced gut dysbiosis and restored the Firmicutes/Bacteroidota balance (P < 0.01). At the genus level, morusin-H significantly reduced the abundances of norank_f_Muribaculaceae, Desulfovibrio, Parabacteroides, and Muribaculum (P < 0.05, P < 0.01, or P < 0.001). At the phylum, family, and genus levels, our findings indicated that morusin-H treatment caused a significant decrease in the abundance of Desulfobacterota, Desulfovibrionaceae, and Desulfovibrio (P < 0.01). Importantly, the abundance of Desulfovibrio was positively correlated with the levels of ALT, AST, TNF-α, IL-1β, and IL-6. Transcriptomic and molecular analyses showed that the therapeutic mechanism of morusin-H involved suppression of the IL-17/TNF signaling pathways and downregulating the mRNA levels of Tlr2, Tlr3, Myd88, Il6, and Cxcl10 (P < 0.05 or P < 0.001), as well as the protein levels of key inflammatory mediators (IL-6, MYD88, and TLR2) (P < 0.001). Conclusion Morusin demonstrates protective effects against LPS-induced acute liver injury, likely through modulation of gut microbiota and suppression of pro-inflammatory factor expression. These findings indicate that morusin exerts its effects through the "microbiota-inflammation-liver" axis, providing a theoretical basis for its use as a multi-target plant-based drug in the treatment of metabolic inflammation-related liver diseases.

Objective:To analyze the clinical features and prognosis of acute B lymphoblastic leukemia (B-ALL) children carrying a TCF3: : PBX1 fusion gene and to evaluate the prognostic value of this gene.Methods:Retrospective cohort study.A total of 2 164 B-ALL children aged 0-18 years diagnosed and treated at 19 pediatric centers from October 2016 to June 2022 were enrolled.They were divided into the positive group and the negative group according to whether they carried a TCF3: : PBX1 fusion gene.The clinical characteristics, treatment response, adverse reactions, and prognosis of the 2 groups of patients were analyzed.The rank sum and Kruskal-Wallis tests were used to compare two and more than two groups of numerical variables, respectively.Fisher′s exact test was used to compare categorical variables.Results:Among the 2 164 patients, 116 (5.4%) were TCF3: : PBX1 positive, of which 70 patients were female, accounting for 60.3%.There were 840 female patients in the TCF3: : PBX1-negative group, accounting for 41.0%.There was a significant difference in the ratio of females between the TCF3: : PBX1-positive and TCF3: : PBX1-negative groups ( P<0.001).No significant difference was observed in age of onset between the two groups( P>0.05).The proportion of bone marrow naive cells [54.00 (14.00, 76.50)% vs.29.00 (3.00, 68.00)%], white blood cell counts [25.30 (10.46, 60.94)×10 9/L vs.9.03 (4.38, 30.73)×10 9/L] and hemoglobin counts [82.00(63.00, 101.00) g/L vs.74.00(60.00, 90.00) g/L] in the TCF3: : PBX1-positive group were significantly higher than those in the negative group at the onset (all P<0.05).In terms of treatment response, the proportion of peripheral blood naive cells on Day 8 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group [2.00 (0, 9.00)% vs.0 (0, 2.00)%, P<0.001].The proportion of minimal residual disease <0.1% on Day 15 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group ( P=0.038).There were no significant differences in cumulative recurrence rate, treatment-related mortality (TRM), and overall survival (OS) between the TCF3: : PBX1-positive group and TCF3: : PBX1-negative group (all P>0.05).The cumulative recurrence risk of TCF3: : PBX1-positive patients was 9.646 times higher than that of ETV6: : RUNX1-positive patients with better prognosis( HR=9.646, 95% CI: 1.026-90.700, P=0.047).There were no significant differences in TRM and OS between TCF3: : PBX1-positive and ETV6: : RUNX1-positive patients (all P>0.05).A significant enrichment of PAX5 mutations was detected in TCF3: : PBX1-positive patients.Among the 7 high-risk TCF3: : PBX1-positive patients in a single center, 4 patients had PAX5 mutations, and this proportion was significantly higher than that in other patients ( P<0.001). Conclusions:B-ALL children carrying a TCF3: : PBX1 fusion gene have a high remission rate and good long-term prognosis after intensive chemotherapy.It is suggesting that TCF3: : PBX1-positive B-ALL patients should be rated at intermediate risk to receive intensive chemotherapy.

Objective:To analyze the clinical features and prognosis of acute B lymphoblastic leukemia (B-ALL) children carrying a TCF3: : PBX1 fusion gene and to evaluate the prognostic value of this gene.Methods:Retrospective cohort study.A total of 2 164 B-ALL children aged 0-18 years diagnosed and treated at 19 pediatric centers from October 2016 to June 2022 were enrolled.They were divided into the positive group and the negative group according to whether they carried a TCF3: : PBX1 fusion gene.The clinical characteristics, treatment response, adverse reactions, and prognosis of the 2 groups of patients were analyzed.The rank sum and Kruskal-Wallis tests were used to compare two and more than two groups of numerical variables, respectively.Fisher′s exact test was used to compare categorical variables.Results:Among the 2 164 patients, 116 (5.4%) were TCF3: : PBX1 positive, of which 70 patients were female, accounting for 60.3%.There were 840 female patients in the TCF3: : PBX1-negative group, accounting for 41.0%.There was a significant difference in the ratio of females between the TCF3: : PBX1-positive and TCF3: : PBX1-negative groups ( P<0.001).No significant difference was observed in age of onset between the two groups( P>0.05).The proportion of bone marrow naive cells [54.00 (14.00, 76.50)% vs.29.00 (3.00, 68.00)%], white blood cell counts [25.30 (10.46, 60.94)×10 9/L vs.9.03 (4.38, 30.73)×10 9/L] and hemoglobin counts [82.00(63.00, 101.00) g/L vs.74.00(60.00, 90.00) g/L] in the TCF3: : PBX1-positive group were significantly higher than those in the negative group at the onset (all P<0.05).In terms of treatment response, the proportion of peripheral blood naive cells on Day 8 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group [2.00 (0, 9.00)% vs.0 (0, 2.00)%, P<0.001].The proportion of minimal residual disease <0.1% on Day 15 in the TCF3: : PBX1-positive group was significantly higher than that in the negative group ( P=0.038).There were no significant differences in cumulative recurrence rate, treatment-related mortality (TRM), and overall survival (OS) between the TCF3: : PBX1-positive group and TCF3: : PBX1-negative group (all P>0.05).The cumulative recurrence risk of TCF3: : PBX1-positive patients was 9.646 times higher than that of ETV6: : RUNX1-positive patients with better prognosis( HR=9.646, 95% CI: 1.026-90.700, P=0.047).There were no significant differences in TRM and OS between TCF3: : PBX1-positive and ETV6: : RUNX1-positive patients (all P>0.05).A significant enrichment of PAX5 mutations was detected in TCF3: : PBX1-positive patients.Among the 7 high-risk TCF3: : PBX1-positive patients in a single center, 4 patients had PAX5 mutations, and this proportion was significantly higher than that in other patients ( P<0.001). Conclusions:B-ALL children carrying a TCF3: : PBX1 fusion gene have a high remission rate and good long-term prognosis after intensive chemotherapy.It is suggesting that TCF3: : PBX1-positive B-ALL patients should be rated at intermediate risk to receive intensive chemotherapy.

Objective·To explore the role and molecular mechanism of GPR87 in regulating the invasion and migration of non-small cell lung cancer(NSCLC).Methods·Bioinformatics methods,including GEO,UALCAN,KM Plotter and other public database analysis platforms,were used to screen candidate genes related to NSCLC invasion and predict their clinical relevance to NSCLC.Eighty NSCLC clinical patient samples and corresponding clinical pathological data were collected from Yichang Central People's Hospital from January 2018 to August 2020.Immunohistochemistry was used to analyze the expression of GPR87 in tumor tissues and the clinical relevance of GPR87 was analyzed.siRNA-GPR87 and pCMV-GPR87-his were transfected into the human lung adenocarcinoma cell line A549 and the human lung squamous cell carcinoma cell line SK-MES-1,to construct cell lines with low and high expression of GPR87.Transwell assay was used to investigate the effect of GPR87 expression on the migration and invasion ability of NSCLC cells.ELISA was used to detect the secretion of MMP7 in the culture supernatant.RT-qPCR was used to detect the mRNA expression levels of GPR87,MMP2,MMP7,MMP9,E-cadherin,N-cadherin,vimentin,snail,twist,RHOA,RHOC,and ROCK1.ELISA was used to detect the secreted protein MMP7.Western blotting was used to detect the protein expression levels of GPR87,MMP9,E-cadherin,vimentin,RHOA,and ROCK1.Results·Bioinformatics analysis of clinical sample data showed that GPR87 was highly expressed in NSCLC.Patients with higher expression of GPR87 had worse clinical stage and were more prone to lymph node metastasis,suggesting that GPR87 might be a key gene for the high invasiveness of NSCLC.Downregulation of GPR87 expression significantly reduced the invasion and migration ability of A549 and SK-MES-1 cells,while overexpression of GPR87 enhanced the invasion and migration ability of A549 and SK-MES-1 cells.Further detection revealed that downregulation of GPR87 led to decreased mRNA expression levels of MMP2,MMP7,MMP9,RHOA,RHOC,and ROCK1,as well as a reduction in the secretion of MMP7 and the protein expression levels of MMP9,RHOA,and ROCK1 in A549 and SK-MES-1 cells.Overexpression of GPR87 increased the mRNA expression levels of MMP2,MMP7,MMP9,RHOA,RHOC,and ROCK1,as well as the secretion of MMP7 and the protein expression levels of MMP9,RHOA,and ROCK1.Regardless of GPR87 knockdown or overexpression,the expression of genes and proteins related to epithelial-mesenchymal transition(EMT)in the cells did not change significantly.Conclusion·High expression of GPR87 is closely related to the high invasiveness of NSCLC.In SK-MES-1 and A549 cells,GPR87 can activate the RHOA/ROCK1 signaling pathway,promote the expression of MMPs,and ultimately promote the invasion and migration of NSCLC.

Objective·To explore the role and molecular mechanism of GPR87 in regulating the invasion and migration of non-small cell lung cancer(NSCLC).Methods·Bioinformatics methods,including GEO,UALCAN,KM Plotter and other public database analysis platforms,were used to screen candidate genes related to NSCLC invasion and predict their clinical relevance to NSCLC.Eighty NSCLC clinical patient samples and corresponding clinical pathological data were collected from Yichang Central People's Hospital from January 2018 to August 2020.Immunohistochemistry was used to analyze the expression of GPR87 in tumor tissues and the clinical relevance of GPR87 was analyzed.siRNA-GPR87 and pCMV-GPR87-his were transfected into the human lung adenocarcinoma cell line A549 and the human lung squamous cell carcinoma cell line SK-MES-1,to construct cell lines with low and high expression of GPR87.Transwell assay was used to investigate the effect of GPR87 expression on the migration and invasion ability of NSCLC cells.ELISA was used to detect the secretion of MMP7 in the culture supernatant.RT-qPCR was used to detect the mRNA expression levels of GPR87,MMP2,MMP7,MMP9,E-cadherin,N-cadherin,vimentin,snail,twist,RHOA,RHOC,and ROCK1.ELISA was used to detect the secreted protein MMP7.Western blotting was used to detect the protein expression levels of GPR87,MMP9,E-cadherin,vimentin,RHOA,and ROCK1.Results·Bioinformatics analysis of clinical sample data showed that GPR87 was highly expressed in NSCLC.Patients with higher expression of GPR87 had worse clinical stage and were more prone to lymph node metastasis,suggesting that GPR87 might be a key gene for the high invasiveness of NSCLC.Downregulation of GPR87 expression significantly reduced the invasion and migration ability of A549 and SK-MES-1 cells,while overexpression of GPR87 enhanced the invasion and migration ability of A549 and SK-MES-1 cells.Further detection revealed that downregulation of GPR87 led to decreased mRNA expression levels of MMP2,MMP7,MMP9,RHOA,RHOC,and ROCK1,as well as a reduction in the secretion of MMP7 and the protein expression levels of MMP9,RHOA,and ROCK1 in A549 and SK-MES-1 cells.Overexpression of GPR87 increased the mRNA expression levels of MMP2,MMP7,MMP9,RHOA,RHOC,and ROCK1,as well as the secretion of MMP7 and the protein expression levels of MMP9,RHOA,and ROCK1.Regardless of GPR87 knockdown or overexpression,the expression of genes and proteins related to epithelial-mesenchymal transition(EMT)in the cells did not change significantly.Conclusion·High expression of GPR87 is closely related to the high invasiveness of NSCLC.In SK-MES-1 and A549 cells,GPR87 can activate the RHOA/ROCK1 signaling pathway,promote the expression of MMPs,and ultimately promote the invasion and migration of NSCLC.

Objective:To explore the influencing factors of seasonal influenza among pregnant woman in Suzhou from 2015 to 2018.Methods:Based on the data of the influenza follow-up cohort of pregnant women in Suzhou from 2015 to 2018, the basic and clinical characteristics of the cohort were described, and the influencing factors of laboratory-confirmed influenza cases in pregnant women were analyzed by unconditional logistic regression.Results:A total of 19 006 pregnant women were recruited, in whom 479 cases of influenza were laboratory confirmed. Influenza A (H3N2) (42.8%) was the main sub-type. In pregnant women with exposure risk in influenza season, unconditional univariate logistic analysis showed that pregnant women or their husbands had registered permanent residence in Suzhou, pregnant women worked as childminder or nanny, had more than 2 permanent residents in the family except themselves, had medical insurance in Suzhou, had fertility insurance in Suzhou, were in the third trimester at the time of enrollment, had cough in the past month, were pregnant for the first time, had children, before and after pregnancy, spent more time outdoors than before, wore masks more often than before and had changed the frequency of gathering were all related to influenza virus infection in pregnant women. Among them, the first pregnancy, increasing the time of outdoor activity, increasing the frequency of wearing masks, and changing the frequency of gathering were important protective factors. Unconditional multivariate logistic regression analysis showed that the number of permanent residents at home was >2 (a OR=1.24, 95% CI: 1.01-1.52) and being in the third trimester, (a OR=1.56, 95% CI: 1.26-1.91) were the risk factors for maternal infection with influenza virus. Conclusion:Pregnant women with a large number of permanent residents and late pregnancy should pay attention to preventing seasonal influenza.

Objective:To understand the reinfection rate of 2019-nCoV in the previously infected population in Suzhou and compare the illness severity and prognosis of the reinfection cases with the first-time infection cases.Methods:A questionnaire survey was conducted in the persons with previous 2019-nCoV infection reported in Suzhou from January 22, 2020 to November 8, 2022 to collect the information about the incidence of reinfection of 2019-nCoV in this population from December 8, 2022 to January 18, 2023. The persons who were infected with 2019-nCoV for the first time were selected by marching the residence, age and gender at ratio of 1∶2 from 2019-nCoV infection community follow-up cohort of Suzhou. By χ2 test, the clinical symptoms and prognosis of the reinfection case and the first-time infection cases were compared. Results:The reinfection rate of 2019-nCoV was 13.01% (147/1 130) in Suzhou. No reinfection was found within 1-6 months after the first-time infection, the rate of reinfection was 10.59% (95/897) in those with interval of 7-12 months between the reinfection and the first-time infection and 45.61% (52/114) in those with the interval ≥24 months. The lowest reinfection rate was 9.09% (1/11) in those who had completed 4 doses of 2019-nCoV vaccination. The main symptoms of the reinfection cases were similar to those of the first-time infection cases. Except for dry cough, nausea/poor appetite and other symptoms, there were significant differences in other clinical symptoms between the two groups ( P<0.05). In the reinfection cases, fever had shorter duration with lower body temperature. The hospital visit rate in the reinfection cases was 4.08% (6/147), lower than that in the cases with the first-time infection (11.56%, 34/294). The time for negative nucleic acid (antigen) test result and recovery from illness after the reinfection were shorter than those after the first-time infection. Conclusions:Reinfection occurred in some people who had been infected with 2019-nCoV. The interval between the reinfection and the first-time infection and the completion of the 4 doses of booster vaccination were the factors influencing the reinfection rate. The hospital visit rate in the reinfection cases was lower than that in the cases with the first-time infection. The reinfection had similar symptoms and shorter illness duration compared with the first-time infection.

Objective:To investigate the clinical features,therapeutic methods,therapeutic efficacy,and prognostic characteristics of older patients with acute myeloid leukemia(AML).Methods:We collected data from 134 older patients with AML treated at Peking University International Hospital between January 2015 and February 2023.White blood cell count,bone marrow primitive cell count,cytogen-etic and molecular characteristics,and European LeukemiaNet(ELN)risk stratification at initial diagnosis were retrospectively ana-lyzed.Patients were assigned into two groups according to treatment plan―high-intensity chemotherapy and low-dose treatment―to determine whether intensive chemotherapy would yield survival benefits during treatment and the factors affecting survival.Results:Among 36 patients treated with high-intensity chemotherapy,22(61.1%)achieved complete response(CR);among 90 treated with low-intensity therapy,46(51.1%)achieved CR;and among 19 treated with azacitidine(AZA)+ venecra(VEN),14(73.7%)achieved CR.Medi-an overall survival(OS)was 15 months for high-intensity chemotherapy and 14.5 months for low-intensity treatment(P=0.226).According to ELN risk stratification,patients in the low,medium,and high risk groups exhibited OS of 18,14,and 9 months,respectively(P=0.009).OS for high-intensity chemotherapy and low-dose therapy was 22 and 15 months in the low-risk group(P=0.745),9 and 15 months in the medium-risk group(P=0.783),and 9 and 8 months in the high-risk group(P=0.739),respectively.Patients in the intensive chemotherapy group(n=36)had an OS of 15 and 17 months(P=0.689)compared with AZA+VEN treatment(n=19).The prognosis of six patients with TP53 mutation was significantly worse than those without the mutation,and the median OS was 2 months and 14 months,respectively(P=0.004).One-and 3-year survival rates for the low-,medium-and high-risk groups were 79%,53%,and 44%,and 41%,20%,and 3%,respectively.Multivariate analysis revealed that high peripheral blood white blood cell count(P=0.034),ELN risk stratification(P=0.002),and complications(P=0.017)were correlated with OS,while treatment intensity,age,sex,and bone marrow primitive cell count were not significantly correlated with OS.Conclusions:High-intensity chemotherapy did not yield a significant survival benefit in older patients with AML;however,this result needs to be confirmed in patients at low risk.Patients with TP53 mutations had a poor prognosis.Multivariate analyses revealed that baseline mo-lecular characteristics,leukocyte count,and comorbidities were more important than treatment intensity in predicting survival among older patients with AML.

【Objective】 To investigate the relationship between hepatitis B virus X （HBx） protein and EGFR promoter， and the role of HBx protein in activating EGFR/PI3K/p-Akt signaling pathway and inhibiting apoptosis. 【Methods】 EGFR promoter plasmids were constructed and the relationship between HBx and EGFR promoters was characterized using a luciferase reporter assay. EGFR-overexpressing trophoblast cells were constructed， and EGFR expression in the overexpressing cells was knocked down using EGFR shRNA. The expression and localization of EGFR/PI3K/p-Akt were detected by Western blotting and confocal laser microscopy. Cell apoptosis was analyzed using flow cytometry. HBV plasmids carrying either full-length HBx or HBx with a deletion mutation （ΔHBx） and HBx plasmids were transfected into two types of trophoblast cells； HBx and PI3K/p-Akt protein expressions were detected by Western blotting. Cell apoptosis was analyzed using flow cytometry. 【Results】 Co-transfection of HBx and EGFR promoter plasmids in JEG-3 and HTR-8/Svneo cells significantly elevated the expression of EGFR promoter driven luciferase compared with the control group （P<0.01）. In EGFR-overexpressing cells， the expression of PI3K/p-Akt was significantly increased （P<0.01）， whereas the apoptosis rate was significantly decreased for JEG-3 cells and HTR-8/Svneo cells （both P<0.01）. These results were reversed in the EGFR-knock down group. When the intracellular HBx protein was expressed in JEG-3 and HTR-8 cells， PI3K/p-Akt protein expression was significantly increased （both P<0.05）， and the proportion of apoptosis was significantly decreased （both P<0.05）. 【Conclusion】 In placental trophoblast cells， HBx protein activates the expression of EGFR by acting on the EGFR promoter， and inhibits the apoptosis of trophoblast cells via the downstream EGFR/PI3K/p-Akt signaling pathway.

Objective To explore the effects of an aerobic exercise of different durations on the binding capacity of NF-E2-related factor 2(Nrf2)/Kelch-like ECH-associated protein-1(Keap1) in skeletal muscle of mice.Methods Thirty C57BL/6J mice were divided into a control(0h) group,an acute exercise for 3 hours (3h) group and an acute exercise for 6 hours(6h) group.The mice ran on treadmill at the speed of 15 m/min for different durations.The mice were sacrificed immediately after exercise and collected skeletal muscles of legs.The high quality fluorescence assay was done to detect the reactive oxygen species(ROS) level in the skeletal muscles of mice.The binding capacity of Nrf2/Keap1 was detected using co-immunoprecipitation.The expression of Nrf2 and Keap1 was analyzed using the Western blotting.Result Compared with group 0h,the Nrf2/Keap1 binding capacity in skeletal muscles of group 3h and 6h decreased significantly(P＜0.05),but the total Nrf2 in skeletal muscles increased significantly(P＜0.05).No significant differences were observed in the expression of total Keapl.The expression of Nrf2 protein of group 3h and 6h increased significantly compared with 0h,with that of group 6h significantly higher than group 3h(P＜0.05).The ROS level in skeletal muscles of group 6h increased significantly compared with group 0h(P＜0.05).Conclusion The effect of acute aerobic exercise on Nrf2/Keap1 binding capacity in skeletal muscle of mice depends on its duration.