Objective: To investigate the protective effects and mechanisms of quercetin on myocardial cell in diabetic rats. Methods: Male Spregue-Dawley rats were randomly divided into four groups: control group, diabetes group, intervention group with low dose quercetin(50 mg/kg gavage), intervention group with high dose quercetin(100 mg/kg gavage). All groups received streptozocin (STZ) by intraperitoneal injection to induce type 1 diabetes mellitus model expect for control group. After gavage for 1 month, cardiac functions were tested by echocardiography. The ratios of cardiomyocyte apoptosis were detected by TUNEL staining. The protein expression of B-cell lymphoma-2 associated X (BAX), B-cell lymphoma-2 (BCL-2), phosphorylated protein kinase B (pAKT), AKT, phosphorylated forkhead box O3 (pFOXO3), FOXO3 were tested by Western blotting. The mRNA levels of Bax and Bcl-2 were tested by quantitative real-time PCR. Results: The cardiac function of diabetes group reduced compared to control group, but ejection fraction and fractional shortening increased in intervention group with low dose quercetin and intervention group with high dose quercetin (P<0.05). The ratios of cardiomyocyte apoptosis in diabetes group were higher than control group (P<0.05), but it decreased in intervention group with low dose quercetin and intervention group with high dose quercetin (P<0.05). There were no obvious changes between intervention group with low dose quercetin and intervention group with high dose quercetin (P>0.05). The mRNA and protein expression of Bax in diabetes group were higher than that in control group (P<0.05), but they reduced in intervention group with low dose quercetin and intervention group with high dose quercetin compared to diabetes group (P<0.05). The mRNA and protein expression of Bcl-2 in diabetes group were lower than control group (P<0.05), but they increased in intervention group with low dose quercetin and intervention group with high dose quercetin compared to diabetes group (P<0.05). The protein expression of pAKT and pFOXO3 in diabetes group were lower than control group (P<0.05), but they increased in intervention group with low dose quercetin and intervention group with high dose quercetin (P<0.05), there were no obvious changes in AKT and FOXO3 between four groups (P>0.05). Conclusion: Quercetin can reduce apoptosis of myocardial cell in diabetic rats through activating AKT/FOXO3 signaling pathway, thus restore the cardiac function.

［Key words］diabetes; quercetin; myocardial cell; apoptosis; AKT/FOXO3 signalling pathway

Objective: To explore the correlation between mean arterial pressure (MAP) and brachial-ankle pulse wave velocity (BaPWV) in patients with type 2 diabetes mellitus（T2DM）. Methods: A total of 979 patients with T2DM were recruited for this study.  Electronic sphygmomanometer was used for measuring blood pressure in the right arms, and MAP was calculated. Based on MAP, patients were divided into low MAP group (MAP<92.4 mmHg, n=327), medium MAP group (92.4 mmHg ≤MAP<102.4 mmHg, n=326) and high MAP group (MAP≥102.4 mmHg, n=326). The BaPWV and ankle-brachial index were measured by Omron arteriosclerosis detector. The correlation between MAP and BaPWV were evaluated by multiple linear regression analysis and Logistic regression analysis. The ROC curve of MAP was drawn to evaluate its predictive value for arterial stiffness in T2DM. Results: BaPWV level was gradually elevated with MAP in T2DM patients (P<0.01). Correlation analysis indicated that MAP was positively correlated with age, body mass index, waist-to-hip ratio, insulin resistance index, triglyceride, and BaPWV. Multiple linear regression demonstrated that MAP was positively associated with BaPWV after adjustment for associated risk factors (P<0.01). Logistic regression analysis showed that MAP was independently associated with BaPWV in T2DM patients after adjustment for other confounders (OR=1.160，95%CI: 1.132-1.188). The area under the ROC curve of MAP for predicting arterial stiffness was 0.786 (95% CI: 0.759-0.811, P<0.01), and take the best cutoff value of 94.4 mmHg (the sensitivity of was 75.0%, and the specificity was 69.2%). Conclusion: MAP is expected to be a simple and practical indicator to predict the risk of atherosclerosis in patients with T2DM.

［Key words］type 2 diabetes mellitus; arterial stiffness; brachialankle pulse wave velocity; mean arterial pressure

Objective: To explore the expression and significance of β-1,4-galactosyltransferase-I (β-1,4-GalT-I) and tumor necrosis factor-α (TNF-α) in dorsal root ganglia (DRG) of rat lumbar facet arthritis (LFA) model. Methods: Adult SD rats were randomly divided into control group and experimental group. For the experimental group, Freund′s complete adjuvant was used to make rat LFA model. Mechanical withdrawal threshold was used to test the change of the mechanical thresholds of the rat foot. The expression of β-1,4-GalT-I and TNF-α in rat DRG and the activation of mitogen activated protein kinase (MAPK) signaling pathway were detected by Western blotting. The distribution and co-localization of β-1,4-GalT-I and TNF-α in DRG of rats were detected by immunofluorescence double labeling. Results: Compared with the control group, the mechanical thresholds of the foot of rats in the experimental group on the 3rd, 5th and 7th days were significantly lower; β-1,4-GalT-I and TNF-α protein levels increased on the 3rd, 5th and 7th days and reached a peak on the 5rd day, and then gradually returned to normal levels in DRG; β-1,4-GalT-I and TNF-α were expressed in DRG neurons and glial cells, and the expression of β-1,4-GalT-I and TNF-α in the 5rd day DRG neurons and glial cells in the experimental group was significantly higher than that in the control group; There was colocalization between β-1,4-GalT-I and TNF-α and MAPK signaling pathways were also activated in DRG of the experimental group. Conclusion: Inflammation of LFA in rats can lead to increased expression of β-1,4-GalT-I and TNF-α in DRG neurons and glial cells, which may be closely associated with the neurological symptoms of lower limbs caused by LFA.

［Key words］lumbar facet arthritis; dorsal root ganglia; β-1,4-galactosyltransferase-I; tumor necrosis factor-α; inflammation

Objective: To study the effects of three main short-chain fatty acids (SCFAs), i.e. sodium acetate, sodium propionate, and sodium butyrate, on 5-fluorouracilinduced inflammatory response in human mononuclear macrophage THP-1 cells and its potential mechanism. Methods: The THP-1 cells was divided into 5 groups: control group,THP-1 cells were not treated; 5-FU group, THP-1 cells were treated with 2.5 mmol/L 5-FU alone for 24 h; 5-FU+sodium acetate group, 5-FU+sodium propionate group and 5-FU+sodium butyrate group, all were pretreated with corresponding 100 μmol/L SCFAs for 24 h, and then 2.5 mmol/L 5-FU were added for 24 h. The qRT-PCR was used to detect the mRNA expression of various inflammation-related factors in cells. ELISA was used to detect the concentrations of IL-1β, IL-6, and IL-10 in the cell supernatant. Western blotting was used to detect the expression of TLR9, NLRP3, Caspase-1, LC3-Ⅱ, Beclin-1, and the expression of NF-κB p65 protein in the nucleus and cytoplasm. Flow cytometry was used to detect cellular reactive oxygen species. Results: Compared with the control group, TLR9, NLRP3, Caspase-1, IL-1β, IL-6, LC3-Ⅱ, Beclin-1, nuclear NF-κB p65 protein expression and reactive oxygen species in 5-FU group were increased significantly(P<0.01). Compared with 5-FU group, the expression of TLR9 in 5-FU+sodium acetate group was greatly reduced(P<0.01); the expression of IL-1β and reactive oxygen species in 5-FU+sodium propionate group were markedly reduced (P<0.01). the expression of IL-1β and nuclear NF-κB p65 protein in 5-FU+sodium butyrate group was greatly reduced (P<0.01), and intracytoplasmic expression was remarkably increased(P<0.01); three kinds of short-chain fatty acid pretreatments could significantly reduce NLRP3, Caspase-1, IL-6, LC3-Ⅱ and Beclin-1 expression(P<0.01), and increase the expression of anti-inflammatory factor IL-10 (P<0.01). Conclusion: Three short-chain fatty acids, sodium acetate, sodium propionate and sodium butyrate, could inhibit the inflammatory response of THP-1 cells induced by 5-FU, inhibit the level of autophagy, and reduce the amount of cellular reactive oxygen generation, which is possible by inhibiting the TLR9/NF-κB/ROS pathway.

［Key words］short-chain fatty acids; 5-fluorouracil; inflammatory reaction; autophagy; reactive oxygen species(ROS); NF-κB

Objective: To explore changes for circRNAs expression profiles in gastric cancer, as well as the changes of competitive endogenous RNA network regulation of circRNAs-miRNAs-mRNAs. Methods: GSE78092, GPL19978 (a combination of GSE83521 and GSE89143), GSE100170, and STAD(unavailable for the public) were selected from the gene expression omnibus（GEO）database. The total of circRNAs were collected from these data. Then through Starbase, TargetScan and other bioinformatics analysis was performed. Results: The total of 734 differentially expressed circRNAs were obtained from gastric cancer tissues and paracancerous tissues, and 17 circRNAs were screened according to P<0.05, log (fold change) >1.5. Then the interaction analysis of circRNAs and microRNAs (miRNAs) using Starbase database was performed again, a total of 354 target miRNAs were found, which were verified by GEO database (GSE23739), 46 miRNAs were finally obtained, and the targeted mRNAs were analyzed using TargetScan database, the kyoto encyclopedia of genes and genomes(KEGG) analysis and gene ontology (GO) analysis showed that target mRNA can promote or inhibit the occurrence and development of gastric cancer mainly in the aspects of transcriptional regulation, RNA metabolism, intracellular signaling cascade and protein localization. Conclusion: The 17 differentially expressed circRNAs formed a competitive endogenous RNA network of circRNAs-miRNAs-mRNAs, which indicated that circRNAs-miRNAs-mRNAs could regulate the occurrence and development of gastric cancer by regulating transcription and RNA metabolism, and could be used as a predictor of gastric cancer and provide a reference for clinical diagnosis and treatment of gastric cancer.

［Key words］circRNAs；gastric cancer；circRNAs expression profiles；biomarker

Objective: To study the neurotoxicity and mechanisms of tetrabromobisphenol A bis（2-hydroxyetyl） ether（TBBPA-DHEE） on zebrafish. Methods:  Zebrafish embryos were randomly divided into five groups: control group, dimethyl sulfoxide(DMSO) group, TBBPA-DHEE exposure groups（0.086, 0.860 and 8.600 mg/L）. After exposure for five days, the mortality, hatching rate, malformation rate, heart rate and body length were observed. Behavioral experiments were used to analyze the spontaneous movement of embryos and the swimming distance and cumulative duration of larvae under light and dark stimulation. The activities of superoxide dismutase（SOD） and glutathione peroxidase（GPx）, the contents of malondialdehyde, Ca²⁺ and γ-aminobutyric acid in larvae were detected by the assay kits. The mRNA expressions of calcium signaling pathway were detected by quantitative real-time PCR method. Results: （1） TBBPA-DHEE exposure could  greatly increase the mortality and malformation of zebrafish embryos/larval, and inhibit the incubation of embryos. The heart rate and body length were decreased remarkably.（2）TBBPA-DHEE exposure could significantly reduce the number of spontaneous movement of zebrafish embryos; 0.860 and 8.600 mg/L TBBPA-DHEE exposure significantly reduced the swimming distances and cumulative mobile durations under dark condition.（3） TBBPA-DHEE exposure could increase the activities of SOD and GPx and the content of malondialdehyde. The content of Ca²⁺ has been increased and the content of gamma-aminobutyric acid has been decreased significantly after TBBPA-DHEE exposure.（4） TBBPA-DHEE exposure could significantly upregulate the mRNA expressions of Cacna1ab, Atp2a1, Atp2a1l, Camk1ga, Ppp3ca, Plcd3a and Prkca in calcium signaling pathways. Conclusion: TBBPA-DHEE could induce the neurotoxicity of zebrafish embryos by enhancing the activities of endogenous antioxidant enzymes and inducing the expressions of related mRNA in calcium signaling pathway.

［Key words］Tetrabromobisphenol A derivative; zebrafish embryos; oxidative stress; calcium signaling pathway; neurotoxicity

Objective: To evaluate the clinical value of serum creatinine-to-cystatin C ratio (CCR) as a serum marker for screening high-risk groups of NAFLD. Methods:  A total of 162 overweight or obese patients and 64 subjects with normal BMI were included in the study. Subjects were divided into three groups according to the CCR level. Spearman correlation, Logistic regression and ROC curve analysis were used to determine the clinical application value of CCR in NAFLD. Results: After grouping CCR based on tertiles, BMI, waist circumference, blood pressure, fasting blood glucose, 2 hours postprandial blood glucose (2hPG), HbA1c, fasting C-peptide, HOMA-IR, triglycerides, AST, ALT, γ-GT, HDL-C, and the prevalence of NAFLD showed significant differences among high, middle and low tertiles (P<0.05). The CCR value of NAFLD patients was significantly lower than that of subjects without NAFLD (P<0.01). CCR was negatively correlated with fasting blood glucose, HbA1c，AST, γ-GT, HOMA-IR, triglycerides, BMI, and waist circumference. After adjusting for age, gender, and glucose and lipid metabolism markers, the risk of NAFLD in the lowest and middle tertiles of CCR were 5.096 times and 2.722 times, respectively (P<0.05). ROC curve analysis showed that the area under the curve of CCR in the male group and female group were 0.865 (95% CI: 0.769-0.932) and 0.806 (95% CI: 0.733-0.866), and the cut points were 94.44 and 91.43, respectively. Conclusion: CCR is closely related to a variety of NAFLD risk factors and can be used as a simple new indicator for early screening of NAFLD.

［Key words］non-alcoholic fatty liver disease; serum creatinine-to-cystatin C ratio; relative muscle mass

Objective: To explore the changes of resting state functional connectivity of dentate nucleus in patients with amnestic mild cognitive impairment (aMCI), and to analyze its correlation with cognitive function. Methods: A total of 34 patients with aMCI were selected as the observation group, and another 34 healthy elderly people were selected as the control group. Data were acquired by restingstate functional magnetic resonance imaging (rs-fMRI) to compare the differences in functional connectivity between the bilateral dentate nuclei and other regions of the whole brain in the two groups; and partial correlation was used to analyze the correlation between functional connectivity and clinical scale scores and biomarker in patients with aMCI. Results: Compared with the control group, the connections between the left dentate nucleus and the left temporal lobe and the right thalamus, the right dentate nucleus and the left superior temporal gyrus and the right posterior cerebellar lobe were enhanced in the observation group (alphasim correction, P<0.05, voxel number> 100). Mini-mental State Examination (MMSE) and verbal fluency test(VFT) were positively correlated with the functional connectivity between left dentate nucleus and right thalamus ( r=0.399, P=0.026; r=0.383, P=0.025), while the functional connectivity between right dentate nucleus and left superior temporal gyrus was negatively correlated with auditory verbal learning for recognition ( r=-0.552, P=0.001) and positively correlated with phosphorylated tau protein ( r=0.357, P=0.045). Conclusion: The specific cerebellar-cortical functional connection was enhanced in the aMCI stage, and this enhancement is related to the cognitive function scale and biomarker.

［Key words］amnestic mild cognitive impairment; resting-state functional magnetic resonance; dentate nucleus; cerebellum; cognition

Objective: To explore the clinical application of plasma cell-free double-stranded DNA (cf-dsDNA) in the diagnosis of primary hepatic carcinomas (PHC). Methods: Sixty-two PHC patients and 41 patients with benign liver disease and 45 healthy controls were recruited. All subjects were required to fill in a questionnaire, and the fasting blood samples were taken from healthy individuals and pre-treatment patients. The plasma cf-dsDNA was extracted by magnetic bead method and the concentration was determined by Qubit 3.0 fluorometer. The serum tumor markers AFP, CA19-9, CA12-5 and CEA levels were measured by electrochemiluminescence. The differences in plasma cf-dsDNA levels in each group of patients and the relationship between plasma cf-dsDNA levels and the clinical characteristics of PHC patients were analyzed, and receiver operating characteristic (ROC) curves were used to analyze the diagnostic efficiency of cf-dsDNA for PHC. Results: The median of preoperative cf-dsDNA levels in PHC patients was significantly higher than those in patients with benign liver disease or healthy controls (H=93.54, P<0.01). In PHC patients, there was no difference in cf-dsDNA levels between age, gender, AFP level, PHC type, and tumor size (all of P>0.05). There was a certain rank correlation between cf-dsDNA levels and TNM stage of PHC patients (r_s=0.311, P=0.013 9). Compared with other tumor markers, cf-dsDNA had higher diagnostic efficiency for PHC (AUC=0.965). According to the cut-off value (>1.912 ng/μL), the positive rate of cf-dsDNA in PHC was 90.32% (56/62), which was significantly higher than other tumor markers (x²= 36.00, P<0.01). Conclusion: Plasma cf-dsDNA was a promising non-invasive marker for PHC diagnosis.

［Key words］cell-free double-stranded DNA； tumor marker； primary hepatic carcinomas； diagnosis

Objective: To investigate the association between small intestinal bacterial overgrowth（SIBO） and the severity of heart failure in chronic heart failure（CHF）patients. Methods: A total of 160 CHF patients admitted to the Department of Cardiology of Zhongshan Hospital were recruited as the CHF group, and 50 nonCHF patients admitted in the same period were selected as the control group. Lactulose hydrogen breath test（LHBT）was used to compare the incidence of SIBO between the CHF group and the control group; in addition, the association between the severity of heart failure（NYHA cardiac function classification）and the positivity of SIBO in patients with CHF was also analyzed. The association between SIBO positivity and clinical characteristics of CHF patients was further analyzed, and the ordered multi-categorical Logistic regression was used to screen the factors associated with the severity of heart failure in CHF patients. Results: ① The positive rate of SIBO in patients with CHF（53.8%）was significantly higher than that in the control group（32.0%， x²=7.214，P<0.01). ② The positive rates of SIBO in NYHA cardiac function class Ⅰ-Ⅳ patients were 20.0%, 36.5%, 66.7% and 71.4%, respectively, with statistically significant differences（x²=16.820，P<0.01). ③ C reactive protein, NT-proBNP and pulmonary artery pressure in SIBO positive group were significantly higher than those in SIBO negative group（all P<0.05). The incidences of jugular vein distention, pulmonary rales and lower limb edema in SIBO positive group were 29.1%, 24.4% and 46.5%, respectively, which were significantly higher than those in SIBO negative group（all P<0.05). ④ The positivity of SIBO was significantly associated with the aggravation of cardiac function class（OR=2.661, 95% CI：1.30-5.45, P=0.007). Conclusion: The positivity of SIBO is associated with CHF, and the incidence of SIBO in CHF patients increases with the severity of heart failure.

［Key words］chronic heart failure; small intestinal bacterial overgrowth; intestinal flora; NYHA functional classification

Objective: To analyze the key components and targets of LiangxueXiaoban Decoction in the treatment of psoriasis vulgaris by network pharmacology and molecular docking, and to explore its possible mechanism. Methods: Traditional Chinese medicine systems pharmacology (TCMSP) and bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine (BATMAN-TCM) were employed to search the effective active components and targets of LiangxueXiaoban Decoction. GeneCards database was employed to search the psoriasis vulgaris related genes. The drug-component-target-disease network and protein-protein interaction (PPI) network was constructed by STRING databaseand Cytoscape 3.7.2 software. Target gene functions and pathways were analyzed by bioinformatics annotation database(Metascape). Finally, Autodock software was applied in verifying the molecular docking between active ingredients and potential protein targets. Results: The key targets of LiangxueXiaoban Decoction in the treatment of psoriasis vulgaris are AKT1, VEGFA, IL-6, JUN and PTGS2, etc. The key KEGG pathways are IL-17 signaling pathway, PI3K-Akt signaling pathway and AGE-RAGE signaling pathway. The molecular docking results showed that diosgenin, kaempferol, luteolin, and baicalein can bind to targets such as PTGS2 and AKT1 with lower binding energy. Conclusion: LiangxueXiaoban Decoction can improve the pathological factors such as inflammatory infiltration of psoriasis, abnormal differentiation of keratinocytes and abnormal new angiogenesis by regulating IL-17 and PI3K/AKT signal pathways, which may contribute to its therapeutic role in psoriasis.

［Key words］network pharmacology; molecular docking; psoriasis; LiangxueXiaoban Decoction； target； signaling pathway

Objective: To prepare conjugates of quantum dots and Staphylococcal protein A (SPA) and figure out the optimal conditions. Methods: The “classical injection method” was used to prepare cadmium telluride quantum dots, while 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) were applied as coupling agents for coupling quantum dots with SPA. A single factor inspection method was adopted to explore the effects of the different amount of EDC, NHS, SPA and the reaction time on the coupling efficiency. Results: A series of conjugates were prepared with different ratios of quantum dots, EDC, NHS, SPA and reaction time. Among them, the fluorescence intensity of the conjugate is the strongest while the ratio of quantum dots to EDC is 1 ∶1 500, the ratio of EDC to NHS is 1 ∶2, and the reaction time is 2 h. Conclusion: When the ratio of cadmium telluride quantum dots to EDC and NHS is 1 ∶1 500 ∶3 000, and the reaction time is 2 h, the cadmium telluride quantum dot-SPA conjugate with better fluorescence intensity could be prepared.

［Key words］quantum dots; Staphylococcal protein A; coupling