ObjectiveTo explore the key molecules regulated by norcantharidin (NCTD) in colon cancer treatment.MethodsWe used cell counting kit-8 and 5-ethnyl-2′-deoxyuridine/Hoechst staining assays to study the effects of NCTD on cell proliferation in colon cancer. Annexin V-fluorescein isothiocyanate/propidium iodide staining was used to evaluate apoptosis, whereas Transwell assays were conducted to evaluate migration and invasion. We performed RNA sequencing to analyze the changes in gene expression after treatment. Differential analysis was performed using differential expression sequencing 2 (Deseq2) in R. Cytoscape was used to construct a competing endogenous RNA (ceRNA) network and Gene Expression Omnibus (GEO) datasets were used to validate sine oculis homeobox homolog 4 (SIX4) expression in colon cancer tissues. Furthermore, the prognostic potential of SIX4 was evaluated using receiver-operating characteristic curves. We conducted an immune infiltration analysis to explore the SIX4 relationship with the immune microenvironment in colon cancer. Finally, SIX4 expression, pan-cancer prognosis, tumor mutation burden (TMB) correlations, microsatellite instability (MSI), and mismatch repair (MMR) were analyzed.ResultsNCTD inhibited colon cancer cell proliferation (P .0001), induced apoptosis (P = .0007), and suppressed the migration and invasion of colon cancer cells. The H19/miR-193b-3p/SIX4 axis was identified as the key ceRNA network involved in the anticancer activity of NCTD. SIX4 is highly expressed in colon cancer tissues, shortening patient survival and affecting immune infiltration. A pan-cancer analysis showed that SIX4 overexpression affects the survival of various cancers. Finally, we correlated SIX4 expression with TMB, MSI, and MMR expression.ConclusionNCTD inhibits the malignant behaviour of colon cancer cells. SIX4 is abnormally expressed in multiple tumor types, significantly affecting the overall survival of patients with cancer, and is a core regulatory target of NCTD in the treatment of colon cancer.

Acute lung injury is an acute and criti-cal respiratory disease with complex pathogenesis,which has a high incidence rate and high mortality.At present,acute lung injury is mainly treated by mechanical ventilation,glucocorticoid anti-inflam-matory and extracorporeal membrane oxygen-ation.Traditional Chinese medicine also shows great potential in improving the clinical manifesta-tions and prognosis of patients.Quercetin is a fla-vonol compound,which can prevent and treat acute lung injury by anti-inflammatory,anti-oxida-tive stress,regulating programmed cell death,anti-bacterial and antiviral.This article summarizes the mechanism of action and research progress of quercetin in the treatment of acute lung injury,aiming to provide reference for subsequent re-search and clinical applications.

Objective:To construct a stable oral squamous cell carinoma(OSCC)cell line overexpressing miR-181a-5p,and to ob-serve the effects of miR-181a-5p on the biological behavior of the cells.Methods:miR-181a-5p lentivirus and the control virus vec-tor were respectively constructed and respectively transfected into OSCC CAL-27 cells.The stablely transfected cells(OE)and con-trol cells(NC)were screened out by purinomycin.The fluorescence of the cells was observed by laser confocal microscopy.The ex-pression of miR-181a-5p and Bcl-2 was detected by qRT-PCR and the expression of target protein Bcl-2 was detected by Western blot.The invasion,migration and proliferation ability of the cells were detected by cell scratch,Transwell assay and clonal formation assay.qRT-PCR was used to detect the expression of the related genes of cell invasion and migration.Results:It was found that the expression level of miR-181a-5p in OE cells was significantly higher than that in NC and untransfected control cells(P＜0.01).The expression of Bcl-2 in OE cells was significantly decreased.The invasion,migration and proliferation of OE cells were decreased.In ad-dition,the expression levels of MMP2,MMP9,Vimentin and Ki67 in OE cells were decreased,while E-cadherin was increased(P＜0.01).Conclusion:Overexpression of miR-181a-5p can significantly inhibit the proliferation,invasion and migration OSCC cells.

Objective:To construct a stable oral squamous cell carinoma(OSCC)cell line overexpressing miR-181a-5p,and to ob-serve the effects of miR-181a-5p on the biological behavior of the cells.Methods:miR-181a-5p lentivirus and the control virus vec-tor were respectively constructed and respectively transfected into OSCC CAL-27 cells.The stablely transfected cells(OE)and con-trol cells(NC)were screened out by purinomycin.The fluorescence of the cells was observed by laser confocal microscopy.The ex-pression of miR-181a-5p and Bcl-2 was detected by qRT-PCR and the expression of target protein Bcl-2 was detected by Western blot.The invasion,migration and proliferation ability of the cells were detected by cell scratch,Transwell assay and clonal formation assay.qRT-PCR was used to detect the expression of the related genes of cell invasion and migration.Results:It was found that the expression level of miR-181a-5p in OE cells was significantly higher than that in NC and untransfected control cells(P＜0.01).The expression of Bcl-2 in OE cells was significantly decreased.The invasion,migration and proliferation of OE cells were decreased.In ad-dition,the expression levels of MMP2,MMP9,Vimentin and Ki67 in OE cells were decreased,while E-cadherin was increased(P＜0.01).Conclusion:Overexpression of miR-181a-5p can significantly inhibit the proliferation,invasion and migration OSCC cells.

Acute lung injury is an acute and criti-cal respiratory disease with complex pathogenesis,which has a high incidence rate and high mortality.At present,acute lung injury is mainly treated by mechanical ventilation,glucocorticoid anti-inflam-matory and extracorporeal membrane oxygen-ation.Traditional Chinese medicine also shows great potential in improving the clinical manifesta-tions and prognosis of patients.Quercetin is a fla-vonol compound,which can prevent and treat acute lung injury by anti-inflammatory,anti-oxida-tive stress,regulating programmed cell death,anti-bacterial and antiviral.This article summarizes the mechanism of action and research progress of quercetin in the treatment of acute lung injury,aiming to provide reference for subsequent re-search and clinical applications.

Objective To analyze the effects of miR-181a-5p overexpression on metabolites in the small intestines of mice with subcutaneous oral cancer by detecting changes in metabolites and metabolic pathways.Methods Three groups were included in study:Control group,negative control and miR-181a-5p overexpression group.To establish a subcutaneous oral cancer model in mice,variously treated cell suspensions were subcutaneously injected into the upper right of the groin in female M-NSG severely immunodeficient mice.Changes in pathology and small intestinal tissues were assessed by HE staining.Changes in mouse body weight were also assessed.Tandem orbitrap mass spectrometry and ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry,were used to examine metabolites in the small intestines.By pre-analyzing the original data and quality rating sample data,XCMS was able to assess which metabolites were different among the groups.To identify unique metabolic pathways,KEGG enrichment analysis was used.Results A total of 170 distinct metabolites were found in the small intestinal tissues of Control and NC groups.Choline metabolism,alanine,aspartate,and glutamate metabolism,GABA synaptic metabolism,glycerophospholipid metabolism,cAMP signaling route,cancer center carbon metabolism,and niacin and niacin amine metabolic pathways were important signaling pathways for metabolite enrichment.In the NC group,16 distinct metabolites with VIP values larger than 2 were found in the small intestines compared with the OE group overexpressing miR-181a-5p.Glycerin phosphorylcholine,palmitic acid,3-hydroxybutyl carnitine,and β-hydroxybutyric acid were among the metabolites that significantly varied.The primary enhanced metabolic pathway was the choline pathway.Conclusions Mouse small intestines underwent slight changes from subcutaneous oral cancer with the greatest effect on metabolites critical for energy metabolism.The choline metabolic pathway was the pathway that selected absolutely metabolites in mouse small intestines with subcutaneous grafts of oral cancer.

Objective To observe the effect of acanthopanax refined polysaccharide(ASPS)on nicotine-induced learning and memory impairment in mice.Methods A total of 48 male mice and 48 females at 6 weeks of age were selected and were divided into two batches for animal experiments:the Morris and the new object recognition batch.Each batch was randomly divided into 6 groups ac-cording to body weight:blank control group,model group,drug positive group,high-dose ASPS group,medium-dose ASPS group,and low-dose ASPS group.Except for the blank group,the remai-ning 5 groups were injected subcutaneously with 0.5mg/kg of nicotine every day for 7 days to prepare a nicotine memory disorder model.After 24 hours of injection of nicotine at the 7th day,the drug posi-tive group was gavaged piracetam for 800 mg/kg,and the high-dose,medium-dose and low-dose ASPS groups were gavaged for 270,90 and 30 mg/kg of ASPS respectively for 7 days.The learning and memory ability of mice was detected by water maze test and new object recognition test,respec-tively.After the two tests,superoxide dismutase(SOD)activity in serum and 5-Hydroxytryptamine(5-HT)content in hippocampal tissue were detected.Results The results of new object recognition experiment showed that the discrimination indexes of the high-,medium-and low-dose ASPS groups were significantly higher than that of the model group(P＜0.01 or P＜0.05).The results of water maze experiment showed that the time to find the platform in the spatial search experiment was signifi-cantly shorter in the high-and medium-dose ASPS groups than that in the model group(P＜0.05).In the positioning voyage test,the number of mouse platform entries in the high-dose ASPS group was significantly more than that in the model group(P＜0.05);the proportions of Ⅲ quadrant routes in the high-and medium-dose ASPS groups were higher than those in the model group(P＜0.05).The high-,medium-and low-dose ASPS groups were significantly higher than those in the model group(P＜0.01);the determination of hippocampal tissue content in mice showed that the content of 5-HT in the high-and medium-dose ASPS groups was significantly higher than that in the model group(P＜0.01 or P＜0.05).Conclusion ASPS can significantly improve the learning and memory ability of nicotine-quitting mice,relieve the damage of hippocampal neurotransmitters,and regulate oxidative stress in vivo.The mechanism may be related to improving the body's antioxidant capacity and regulating hippocampal neurotransmitter levels.

Objective To observe the effect of acanthopanax refined polysaccharide(ASPS)on nicotine-induced learning and memory impairment in mice.Methods A total of 48 male mice and 48 females at 6 weeks of age were selected and were divided into two batches for animal experiments:the Morris and the new object recognition batch.Each batch was randomly divided into 6 groups ac-cording to body weight:blank control group,model group,drug positive group,high-dose ASPS group,medium-dose ASPS group,and low-dose ASPS group.Except for the blank group,the remai-ning 5 groups were injected subcutaneously with 0.5mg/kg of nicotine every day for 7 days to prepare a nicotine memory disorder model.After 24 hours of injection of nicotine at the 7th day,the drug posi-tive group was gavaged piracetam for 800 mg/kg,and the high-dose,medium-dose and low-dose ASPS groups were gavaged for 270,90 and 30 mg/kg of ASPS respectively for 7 days.The learning and memory ability of mice was detected by water maze test and new object recognition test,respec-tively.After the two tests,superoxide dismutase(SOD)activity in serum and 5-Hydroxytryptamine(5-HT)content in hippocampal tissue were detected.Results The results of new object recognition experiment showed that the discrimination indexes of the high-,medium-and low-dose ASPS groups were significantly higher than that of the model group(P＜0.01 or P＜0.05).The results of water maze experiment showed that the time to find the platform in the spatial search experiment was signifi-cantly shorter in the high-and medium-dose ASPS groups than that in the model group(P＜0.05).In the positioning voyage test,the number of mouse platform entries in the high-dose ASPS group was significantly more than that in the model group(P＜0.05);the proportions of Ⅲ quadrant routes in the high-and medium-dose ASPS groups were higher than those in the model group(P＜0.05).The high-,medium-and low-dose ASPS groups were significantly higher than those in the model group(P＜0.01);the determination of hippocampal tissue content in mice showed that the content of 5-HT in the high-and medium-dose ASPS groups was significantly higher than that in the model group(P＜0.01 or P＜0.05).Conclusion ASPS can significantly improve the learning and memory ability of nicotine-quitting mice,relieve the damage of hippocampal neurotransmitters,and regulate oxidative stress in vivo.The mechanism may be related to improving the body's antioxidant capacity and regulating hippocampal neurotransmitter levels.

Objective:To explore the current status of mental workload and its relationship with work engagement and fatigue, as well as the impact path among the three.Methods:Clinical nurses from six tertiary hospitals including Shanghai Traditional Chinese Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, etc. were selected to conduct a cross-sectional survey using convenience sampling method from February to March 2020. They were investigated by general information questionnaire, NASA Task Load Index (NASA-TLX), Utrecht Work Engagement Scale (UWES), and Fatigue Scale-14 (FS-14).Results:The total mental workload score of 776 clinical nurses was 76.50(69.00, 84.00). Single factor analysis showed that clinical nurses of different ages, marriage and childbirth status, education level, professional title and working years had different mental workload ( Z = -2.61, H values were 10.22-22.41, all P<0.01). Bivariate analysis revealed that the mental workload of clinical nurses was positively correlated with work engagement ( r = 0.27, P<0.01) and fatigue ( r = 0.23, P<0.01), and work engagement and fatigue were negatively correlated ( r = -0.23, P<0.01). Mediation effect analysis demonstrated that mental workload had a positive predictive effect on fatigue ( β = 0.39, P<0.01) and work engagement ( β = 0.35, P<0.01); the suppressing effect of work engagement between mental workload and fatigue, the absolute value of the ratio of the suppressing effect to the direct effect was |-0.17/0.39|. Conclusions:The mental workload of clinical nurses is at relatively high level. Hospital administrators can partially improve the fatigue state of clinical nurses with high mental load through the adjustment effect of work engagement.

Objective To study the immunomodulatory effect of polysaccharides (CRPS25-Ⅱ) derived from Chroogomphus rutilus on mouse mononuclear macrophages, RAW264.7 cells. Methods RAW264.7 cells were resuspended and cultured, cell suspension was prepared. The blank control group and CRPS25-Ⅱ groups with different mass concentrations (1, 20, 40, 80 and 160 μg/ml) were set up. MTT assay was used to determine the cytotoxicity of CRPS25-Ⅱ on RAW264.7 cells. RT-PCR was used to detect the effects of CRPS25-Ⅱ on the secretion of immune regulatory factors IL-6 and TNF-α from RAW264.7 cells. Western blot was used to detect the effects of CRPS25-Ⅱ on the expression of p-P65 protein in NF-κB pathway of RAW264.7 cells. Results The results showed that CRPS25-Ⅱ (1−160 μg/ml) had no obvious cytotoxicity. CRPS25-Ⅱ (1−160 μg/ml) increased the secretion of cytokines, and thus promoted the mRNA expression of IL-6 and TNF-α. CRPS25-Ⅱ increased the phosphorylation of p-P65 protein and activated the NF-κB signaling pathway, and thus promoted the immune regulation of cells. CRPS25-Ⅱ (1−160 μg/ml) could increase the p-P65 protein, and the promoting effects of CRPS25-Ⅱshowed an upward trend in the concentration range of 1−40 μg/ml and gradually weakened in the concentration range of 40−160 μg/ml. Conclusion Polysaccharides derived from chroogomphus rutilus had no cytotoxicity to mouse macrophages, and could promote the secretion of inflammatory factors IL-6 and TNF-α and activate the NF-κB signaling pathway, thus playing an immunomodulatory role.