In this work, starting from 4-hydroxycoumarin, three series of 22 coumarin derivatives, among which 8 have not been reported in the literature, were synthesized and their in vitro anti-inflammatory activities and mechanisms of action were preliminarily investigated using mouse macrophage model. The results showed that most of the derivatives could significantly inhibit the production of pro-inflammatory factor NO, with compounds 2e, 2f, 2g, 2h, 2i, 2j, 4e, and 4f showing better anti-inflammatory activity than the positive control drug dexamethasone. Further experiments showed that compounds 2h and 4f significantly inhibited the production of pro-inflammatory factors IL-6, TNF-α and IL-1β in RAW264.7 macrophages, and could, therefore, be used as lead compounds for further studies.

Objective:To investigate the application value of a new clinical probation log designed based on the subjective-objective-assessment-plan (SOAP) structured medical record.Methods:Quantitative and qualitative studies were conducted among 97 students in the autumn semester to evaluate the effect of the new clinical probation log based on the SOAP structured medical record on their academic performance and clinical thinking. SPSS 26.0 was used to perform the t-test and ANOVA. Results:The students using the new version had a significantly better score (96.29±1.38) than those using the old version (93.53±1.60) ( P<0.001), while the multivariate analysis showed that the students who first used the old version and then switched to the new version had a significantly better improvement in the score compared with those who first used the new version and then switched to the old version ( P<0.001). The qualitative interview showed that it was necessary to use handwritten internship log, and compared with the old version, the new version could better promote the ordered clinical thinking of students. The teacher comments could give feedbacks, and all the students interviewed thought that the new version held promise for clinical application. Conclusions:The clinical probation log based on the SOAP structured medical record can help to improve the effectiveness of probation and cultivate clinical thinking ability, and thus it holds promise for application in clinical probation teaching.

Objective To investigate the protective effect of AKAP12/PCK6 signaling pathway on podocyte autophagy in diabetic kidney disease(DKD).Methods A total of 40 C57/B6 AKAP12 KO(AKAP12-/-)mice and wild-type(WT)littermates were randomly divided into four groups,with 10 mice in each group:Ctrl+WT group,Ctrl+AKAP12-/-group,DKD+WT group and DKD+AKAP12-/-group.Primary podocytes were obtained from AKAP12-/-mice and WT mice,and exposed to high glucose(HG,30 mmol/L)or mannitol for 24 h to investigate the mitosis and autophagy of podocytes.The primary podocytes were divided into Mannitol+WT,G+WT,Mannitol+AKAP12-/-and HG+AKAP12-/-.In addition,AKAP12-/-podocytes were transfected with sh-PCSK6 to knockdown the expression of PCK6.Mito-Tracker staining was used to analyze the morphology of mitochondria in podocytes.The expressions of mitotic proteins(FIS1 and DRP1),mitochondrial autophagy(PINK1 and Parkin)and autophagy-related proteins(LC3 and p62)were analyzed by Western blot.Results Compared with HG+WT group,the number of single mitochondria and the expression of FIS1,DRP1,PINK1,Parkin,LC3Ⅱ and p62 proteins increased,while the average branch length of mitochondria decreased in HG+AKAP12-/-group(P＜0.05).Compared with HG+sh-NC group,the number of single mitochondria and the expression of FIS1,DRP1,PINK1,Parkin and LC3Ⅱ proteins decreased significantly in HG+sh-PCSK6 group(P＜0.05 or P＜0.01),while the average branch length of mitochondria increased significantly(P＜0.05).Conclusions AKAP12/PCSK6 signaling pathway mediates the regulation of mitochondrion division and mitochondrion autophagy in podocytes under HG environment.

Objective:To explore the relationship among sleep phenotypes, attention deficit and hyperactivity impulsivity (ADHD) symptoms and cognitive information processing in children with ADHD.Methods:A total of 244 children with ADHD aged 6-12 were selected from December 2021 to December 2022.Swanson, Nolan and Pelham rating scale Ⅳ(SNAP-Ⅳ) was used to evaluate the core symptoms of ADHD.Sleep disturbance scale for children (SDSC) was used to evaluate six sleep phenotypes((disorders in initiating and maintaining sleep (DIMS), sleep breathing disorders(SDB), sleep-wake transition disorders(SWTD), disorders of arousal(DA), disorders of excessive somnolence (DOES), and Nocturnal hyperhidrosis(SHY)). Das-Naglieri cognitive function assessment system (DN-CAS) was used to evaluate the cognitive information processing (planning, simultaneous processing, attention and successive processing). Descriptive statistical analysis, Spearman correlation analysis, and mediation analysis were conducted by SPSSAU 23.0 and Zstats software, respectively.Results:Correlation analysis showed that DIMS, SDB, SWTD, DA and DOES were significantly and positively correlated with attention deficit ( r=0.190-0.349, all P<0.01).DIMS(2.14(1.71, 2.57)), SWTD(1.67(1.33, 2.00)) and SHY(2.00(1.50, 3.00)) were significantly positively correlated with hyperactive impulsivity (1.44(1.00, 2.00))( r=0.193, 0.242, 0.133, P<0.05). Attention deficit(1.78(1.44, 2.33)) was significantly and negatively correlated with successive processing(105.00(96.00, 112.00)) ( r=-0.127, P<0.05). The results of multiple linear regression analysis showed that after controlling for sex, age, verbal IQ and operational IQ, DIMS ( β=0.152, P<0.05) and SWTD ( β=0.178, P<0.05) had significant positive predictive effects on hyperactive impulsivity symptoms. DOES ( β=0.259, P<0.01) had significant positive predictive effects on attention deficit symptoms. Attention deficit ( β=-0.183, P<0.05) had a significant negative predictive effect on successive processing. Mediation effect analysis showed that attention deficit played a complete mediating role between DOES and successive processing(effect=-0.179, Bootstrap 95% CI=-0.196--0.110). Conclusion:Different sleep phenotypes are associated with ADHD core symptoms and different dimensions of cognitive information processing processes. DOES indirectly affects successive processing capability by attention deficit symptoms.

Objective To investigate the expression of miR-204 and silence information regulator 1(SIRT1)in colorectal cancer and their clinical value.Methods Cancer tissue specimens and paracancer tissue specimens of 60 patients with colorectal cancer treated in Department of Gastrointestinal Surgery of Affiliated Hospital of Jiaxing University from May 2018 to June 2020 were collected as study objects.Real time fluorogenic quantitative polymerase chain reaction was used to detect the gene expression of miR-204 and SIRT1,and the correlation between miR-204 and SIRT1 gene expression was compared by Pearson analysis.Immunohistochemical SABC method was used to detect SIRT1 protein expression,and relationship between different SIRT1 protein expression and clinicopathological features was compared.Kaplan-Meier method was used to analyze the survival difference of colorectal cancer patients with different SIRT1 protein expression.Results The mRNA expression of miR-204 gene in cancer tissues was significantly lower than that in paracancer tissues(P<0.05),and the mRNA expression of SIRT1 gene was significantly higher than that in paracancer tissues(P<0.05).Pearson correlation analysis showed that miR-204 was negatively correlated with SIRT1 mRNA expression in both paracancer tissues and cancer tissues(r=-0.647,-0.737,P<0.05).The expression of SIRT1 protein was correlated with the differentiation level,invasion level,lymph node metastasis and TNM stage of colorectal cancer(P<0.05),but not with patient age,gender,tumor size and tumor site(P>0.05).Kaplan-Meier analysis showed that the survival rate of patients with positive expression of SIRT1 in cancer tissues was significantly lower than that of patients with negative expression of SIRT1(χ2=5.001,P=0.025).Conclusion The expression of miR-204 is down-regulated and SIRT1 expression is up-regulated in colorectal cancer tissues,which may jointly promote the metastasis and invasion of colorectal cancer and affect the prognosis of patients through mutual influence.

Objective:To investigate the differences in multi-b-value apparent diffusion coefficient (ADC) and exponential apparent diffusion coefficient (eADC) between hepatocellular carcinoma (HCC) and paracancerous liver tissue, distant cancerous liver tissue, and background liver tissues by ultra-high field 3.0T diffusion-weighted (DWI) MRI imaging.Methods:Sixty-eight consecutive HCC cases confirmed by surgical pathology from January 2018 to October 2021 were enrolled and divided into a cirrhosis ( n=39) and a non-cirrhosis group ( n=29) according to the presence or absence of cirrhosis.The average ADC and eADC of liver tissues of paracancerous (including proximal and distal), distant cancerous, and background were measured by DWI images with diffusion sensitivity factors (b) of 50, 100, 400, 600 s/mm 2, and 1 000 s/mm 2, respectively. The Kruskal-Wallis H test and Bonferroni method were used to test the differences between the measured values of the five tissues. The statistical differences were used to evaluate the diagnostic efficacy of the five tissues by parametric receiver operating characteristic (ROC) curve and area under the curve (AUC). Results:The comparison of average ADC and eADC among five types of tissues in the liver cirrhosis group showed that the average ADC and eADC measured at b values of 50, 100, 400, and 600 s/mm 2 had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, distant cancerous, and background liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and proximal paracancerous tissue. The average ADC and eADC in the non-cirrhosis group had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue measured at b values of 50, 100, and 400 s/mm 2, respectively. The average ADC and eADC measured at b=600 s/mm 2 showed statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, and distant cancerous liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and distal paracancerous, and distant cancerous liver tissue. The average ADC and eADC in the cirrhosis group had no statistically significant difference between the proximal paracancerous and the distant cancerous, as well as the background liver tissue measured at b-values of 50, 100, 400, 600, and 1 000 s/mm 2, respectively (adjusted P>0.005), while there were statistically significant differences (adjusted P<0.005) in the average ADC values in the non-cirrhosis group between the proximal paracancerous and the distant paracancerous and background liver tissues at b=50 s/mm 2, as well as the average ADC and eADC values between the proximal paracancerous and the distant liver tissues at b=100 s/mm 2. The average ADC and eADC values measured in the cirrhosis group and non-cirrhosis group had no statistically significant difference between the distant paracancerous, distant cancerous, and background liver tissue (adjusted P>0.005). The efficacy of average ADC and eADC in distinguishing five types of tissues (cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue) showed that in the cirrhosis group, the diagnostic efficacy was best at b=50 s/mm 2. The area under the ROC curve (AUC) of average ADC was 0.815, 0.828, 0.855, and 0.855, respectively, and the AUC of average eADC was 0.815, 0.830, 0.856, and 0.855, respectively. The diagnostic efficacy was best in the non cirrhosis group at b=100 s/mm 2, with average ADC AUCs of 0.787, 0.823, 0.841, and 0.821, and average eADC AUCs of 0.836, 0.874, 0.893, and 0.873, respectively. The AUC of the average ADC in the non-cirrhosis group for distinguishing between proximal paracancerous and distant cancerous liver tissues, as well as proximal paracancerous and background liver tissues, with b=50 s/mm 2, were 0.605 and 0.604, respectively. The average AUC of ADC and eADC for distinguishing between proximal paracancerous and distant liver tissues with b=100 s/mm 2 were 0.619 and 0.620, respectively. Conclusion:The average ADC and eADC measured by multiple b-values are helpful in distinguishing HCC from proximal paracancerous, distal paracancerous, distant-cancerous, and background liver tissues in patients with cirrhosis and non-cirrhosis, while the average ADC and eADC at b=50 s/mm 2 and 100 s/mm 2 exhibit differences between the proximal paracancerous from the distant cancerous liver tissue and background liver tissue in patients with non-cirrhosis.

Objective To screen small molecule inhibitors of Fusobacterium nucleatum （Fn） based on commercially available compound libraries, and investigate their anti-colorectal cancer activities under Fn intervention in order to obtain novel anti-colorectal cancer lead compounds. Methods The promotion of colorectal cancer proliferation on organoid was validated by Fn. Secondly, the effects of anti-Fn compounds on their in vitro anticancer activity under Fn’s co-incubation with colorectal cancer HCT116 cell were comparative investigated. Finally, in vivo anticancer efficacy of highly active compounds on nude mouse colon cancer HCT116 transplanted tumor under the intervention of Fn was evaluated by gavage. Results Fn could significantly promote the proliferation of rectal cancer organoids. 9 anti-Fn active compounds could significantly enhance their in vitro anticancer activity under Fn’s co-incubation with HCT116 cells. Methotrexate had the strongest anti-cancer activity with IC₅₀ as 0.03 μmol/L. The combined use of methotrexate (0.5 mg/kg) and PD-1 (5.0 mg/kg) had a stronger anti-tumor effect than their standalone use. Conclusion As new small molecule inhibitor of Fn, methotrexate exhibited good in vitro and in vivo anti-colorectal cancer activity against HCT116 cells and nude mouse xenografts under Fn intervention, which showed the foundation for subsequent structural optimization, and could be expected to expand the new indications of methotrexate.

The experimental teaching of psychology is not only an important course in the undergraduate and graduate education of applied psychology, but also can assist the practical teaching of quite a few other subjects. It clearly divides three basic functions of experimental psychology teaching center: teaching, scientific research and service, which play an important role in facilitating teachers and students to understand corresponding courses. At the same time, it is discussed and proposed to strengthen the internal and external scientific linkage of the experimental center under the network background, and put forward the use of Internet technology, in order to improve the scientific use of the experimental teaching center, and reflect its maximum value, thereby achieving the purpose of university laboratory joint construction and serve the university to cultivate innovative and compound personnels.

OBJECTIVE: To explore the mechanisms of large-conductance calcium-activated potassium channel (BKCa) involved in inflammatory response in sepsis. METHODS: The serum levels of BKCa were measured by enzyme-linked immunosorbent assay (ELISA) in patients with sepsis (28 cases), patients with common infection (25 cases) and healthy people (25 cases). The relationship between levels of BKCa and acute physiology and chronic health evaluation II (APACHE II) were analyzed. Cultured RAW 264.7 cells were stimulated by lipopolysaccharide (LPS). In some experiments, a cell model of sepsis was constructed using Nigericin as the second stimulus signal. The mRNA and protein expressions of BKCa in RAW 264.7 cells stimulated with LPS (0, 50, 100, 1 000 μg/L) were measured by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting. RAW 264.7 cells were transfected with small interfering RNA of BKCa (siRNA-BKCa), and the levels of caspase-1 precursor (pro-caspase-1), interleukin-1β precursor (pro-IL-1β) in cell, and the levels of caspase-1 p20, IL-1β p17 of cell culture medium, and NOD-like receptor protein 3 (NLRP3), nuclear factor-κB (NF-κB) were measured by Western blotting. The apoptosis were detected by staining with propidium iodide (PI), the release rate of lactate dehydrogenase (LDH) were measured, and the expression of apoptotic protein Gasdermin D (GSDMD) was measured by Western blotting to evaluate the effect of silencing BKCa on cell pyrosis. RESULTS: The level of serum BKCa in patients with sepsis was significantly higher than that in patients with common infection and health peoples (ng/L: 165.2±25.9 vs. 102.5±25.9, 98.8±20.0, both P < 0.05). In addition, the level of serum BKCa in patients with sepsis was significantly positively correlated with APACHE II score (r = 0.453, P = 0.013). LPS could construct a sepsis cell model by which LPS could promote BKCa expression in mRNA and protein with a concentration-dependent manner. The mRNA and protein expressions of BKCa in the cells stimulated by 1 000 μg/L LPS were significantly higher than that in the blank group (0 μg/L) [BKCa mRNA (2^-ΔΔCt): 3.00±0.36 vs. 1.00±0.16, BKCa/β-actin: 1.30±0.16 vs. 0.37±0.09, both P < 0.05]. Compared with the control group, the ratios of caspase-1 p20/pro-caspase-1 and IL-1β p17/pro-IL-1β in the model group were significantly increased (caspase-1 p20/pro-caspase-1: 0.83±0.12 vs. 0.27±0.05, IL-1β p17/pro-IL-1β: 0.77±0.12 vs. 0.23±0.12, both P < 0.05), however, transfection of siRNA-BKCa induced the decrease both of them (caspase-1 p20/pro-capase-1: 0.23±0.12 vs. 0.83±0.12, IL-1β p17/pro-IL-1β: 0.13±0.05 vs. 0.77±0.12, both P < 0.05). Compared with the control group, the number of apoptotic cells, LDH release rate and GSDMD expression in the model group were significantly increased [LDH release rate: (30.60±8.40)% vs. (15.20±7.10)%, GSDMD-N/GSDMD-FL: 2.10±0.16 vs. 1.00±0.16, both P < 0.05], however, transfection of siRNA-BKCa induced the decrease both of them [LDH release rate: (15.60±7.30)% vs. (30.60±8.40)%, GSDMD-N/GSDMD-FL: 1.13±0.17 vs. 2.10±0.16, both P < 0.05]. The mRNA and protein expressions of NLRP3 in sepsis cells were significantly higher than those in the control group [NLRP3 mRNA (2^-ΔΔCt): 2.06±0.17 vs. 1.00±0.24, NLRP3/GAPDH: 0.46±0.05 vs. 0.15±0.04, both P < 0.05]. However, the expression of NLRP3 after siRNA-BKCa transfection was significantly lower than that in model group [NLRP3 mRNA (2^-ΔΔCt): 1.57±0.09 vs. 2.06±0.17, NLRP3/GAPDH: 0.19±0.02 vs. 0.46±0.05, both P < 0.05]. Compared with the control group, the NF-κB p65 nuclear transfer of sepsis cell were significantly increased (NF-κB p65/Histone: 0.73±0.12 vs. 0.23±0.09, P < 0.05). However, the NF-κB p65 expression in the nucleus were decreased after siRNA-BKCa transfection (NF-κB p65/Histone: 0.20±0.03 vs. 0.73±0.12, P < 0.05). CONCLUSIONS BKCa is involved in the pathogenesis of sepsis, and its possible mechanism is to activate NF-κB/NLRP3/caspase-1 signaling pathway to induce inflammatory factor production and cell death.
Humans ; Histones ; Caspase 1 ; Large-Conductance Calcium-Activated Potassium Channels ; Lipopolysaccharides ; NF-kappa B ; NLR Family, Pyrin Domain-Containing 3 Protein ; L-Lactate Dehydrogenase ; Sepsis ; RNA, Small Interfering ; Caspases

Glioblastoma (GBM), characterized by rapid progression, easy recurrence and treatment resistance, is the most aggressive and lethal tumor of the central nervous system. Adenovirus E1A binding protein p300 (p300) serves rich functions as transcriptional coactivator and lysine acetyltransferase. Studies have shown that p300 plays an important role in the occurrence, proliferation, invasion and resistance to chemoradiotherapy of GBM. This paper reviews the structure and function of p300, and systematically expounds its various ways participating in GBM development and its translational perspectives, so as to provide references for GBM study.