Objective To investigate the effect of β3-adrenoreceptor (β3-AR)overexpression on cardiac hypertrophy. Method Sprague-Dawley rat neonatal ventricular cardiomyocytes (NRVMs) were isolated and cul-tured in vitro.The infection of lentiviruswas examined after cardiomyocytes were infected with lentivirus at differ-ent multiplicity of infection (MOI) of 20、50、80 and 100. Fluorescence microscopy was used to observe the ex-pression of GFP and to confirm the best MOI for lentivirus infection. Following the enforced expression of β3-AR by lentivirus, 2uM norepinephrine (NE) was used to treatment the infected cardiomyocytes for 48h. Expressions of β3-AR、c-myc and c-fos protein in cardiomyocytes were detected by Western Blot. Results The results of fluorescence microscopy indicated that the best MOI was 50. The protein level of β3-AR was significantly in-creased in β3-AR overexpression group compared with the control group and the NE treatment group (P < 0.05). Following the treatment of NE , the expressions of c-myc and c-fos were also significantly increased in β3-AR overexpression group compared with the control group (P < 0.05). Conclusion Over-expression of β3-AR can aggravate cardiomyocyte hypertrophy.

BACKGROUND:Recombinant adeno-associated virus serotype 9 has a high affinity in myocardial tissue, and the expression of recombinant adeno-associated virus serotype 9-enhanced green fluorescent protein (rAAV9-eGFP) in the aorta of atherosclerosis mice is not clear. OBJECTIVE:To explore the optimal time point of rAAV9-eGFP expression in the aorta of atherosclerosis mice. METHODS:Atherosclerosis model was established with high-fat diet in 30 ApoE-/-mice for 16 weeks. Among them, 25 mice were injected with 5.0×1011 vg (virus genomes) rAAV9-eGFP through the tail vein, while the remaining 5 mice were injected with saline, serving as the control group. The virus-transfected mice were kil ed at 14, 21, 28, 35 and 60 days after transfection, and aortic tissue was harvested. The expression of enhanced green fluorescent protein was detected with laser scanning confocal microscope. Western blot assays were used to detect the expression of enhanced green fluorescent protein in aorta. The expression of enhanced green fluorescent protein in vivo was observed and the optimal expression time point was determined. RESULTS AND CONCLUSION:rAAV9-eGFP effectively transfected the aorta of atherosclerosis mice, enhanced green fluorescent protein was expressed in aortic tissue, and the expression intensity increased gradual y with the increasing transfection time. The highest expression level was found at 35 days after transfection and then maintained stable at 60 days. There were significant differences at different time points after transfection (P<0.001). These data indicate that rAAV9-eGFP can be effectively expressed in the aorta of atherosclerosis ApoE-/-mice and rAAV9-eGFP can be regarded as the optimal vector in the treatment of atherosclerosis.

Objective To explore the expression profiles of circulating microRNA(miRNA)and potential markers for the diagnosis of adult fulminant myocarditis(FM). Methods The expression profiles of circulating miRNA were determined by microarray analysis and verified by real-time quantitative PCR.The key role of circulating miRNA in FM was determined via KEGG pathway enrichment.The correlations between miRNA and cardiac function parameters in patients with FM were analyzed.The receiver operating characteristic(ROC)curve was established to evaluate the sensitivity and specificity of circulating miRNA in the diagnosis of FM. Results Compared with healthy controls,the FM patients had up-regulated expression levels of miR-29b(t=18.925,P<0.001)and miR-125b(t=5.981,P=0.029)in the plasma.After treatment,the expression levels of miR-29b(t=12.943,P<0.001)and miR-125b(t=14.016,P<0.001)were significantly down-regulated.KEGG pathway enrichment showed that the targets of miR-29b were involved in inflammatory response and apoptosis pathways.The results of cell proliferation and apoptosis assay demonstrated the transfection of miR-29b mimic had a more significant inducing effect on cardiomyocyte apoptosis than that of miR-125b mimic(χ ²=6.168,P=0.047),whereas there was no significant difference in the inhibition of cell proliferation between the two groups(χ²=1.452,P=0.417).The expression levels of miR-29b and miR-125b were negatively correlated with left ventricular ejection fraction(r=-0.67,P=0.071;r=-0.49,P=0.003).They were positively correlated with cardiac troponin I level(r=0.61,P=0.019;r=0.52,P=0.016),interferon β level(r=0.42,P=0.014;r=0.36,P=0.021),and myocardial edema area(r=0.86,P=0.005;r=0.73,P=0.013).The ROC curve analysis demonstrated that miR-29b had higher sensitivity for the diagnosis of FM(93.6% vs.89.2%;t=0.896,P=0.795)and specificity(72.4% vs.59.6%;t=9.478,P=0.002)than miR-125b. Conclusion The circulating miR-29b may be a potential biomarker for the diagnosis of FM.
Adult ; Biomarkers/metabolism* ; Circulating MicroRNA/metabolism* ; Humans ; MicroRNAs/metabolism* ; Myocarditis/diagnosis* ; Stroke Volume ; Ventricular Function, Left

BACKGROUND:The formation of atherosclerotic lesions in apolipoprotein E knockout mice is similar to that of human systemic atherosclerosis, and apolipoprotein E knockout mice are ideal animals for current establishment of atherosclerosis models.
OBJECTIVE:To research the pathological process of atherosclerosis in apolipoprotein E knockout mice aged different weeks, and to explore the effect of different diets on the occurrence and development of atherosclerosis in apolipoprotein E knockout mice.
METHODS:Male apolipoprotein E knockout mice aged 8 weeks old were randomly divided into two groups, and fed with high fat diet and normal diet, respectively, for 8, 12, 16, 20, and 24 weeks.
RESULTS AND CONCLUSION:Serological detection revealed that serum total cholesterol, triglycerides and low density lipoprotein levels were significantly higher in different weeks of mice of high fat diet group than in the normal diet group (P<0.05), in a time-dependent manner. Gross and frozen oil red O staining showed that atherosclerotic plaque area of lumen was significantly larger in the high fat diet group than in the normal diet group (P<0.05), in a time-dependent manner. At this time, significant differences in plaque area of lumen at each week were detected between both groups (P<0.05). Apparent lipid plaque was visible in aorta at 16 weeks of high fat diet in mice. Results demonstrated that apolipoprotein E knockout mice of atherosclerosis were successful y established. The formation of lipid streaks and fiber hyperplasia was faster in high fat diet group than in the normal diet group.

Objective To evaluate in vitro transfection of anti-nuclear factor-κB ( NF-κB) ribozyme gene to human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) mediated by recombinant adeno-associated virus 9 carrying enhanced green fluorescent protein gene ( rAAV9-EGFP-R65 ) and to study their effects on cell proliferation and NF-κB P65 expression.Methods HUVECs and HASMCs were respectively transfected with rAAV9-EGFP-R65 at different multiplicity of infection ( MOI=1 ×105 , 1 ×106 and 1×107).The expression of EGFP was observed with fluorescence microscopy .Flow cytometry was performed to evaluate the transfection efficiency .Alamar Blue assay was used to measure the proliferation of the transfected cells.Western blot was used to detect NF-κB P65 expression .Results The fluorescence intensity was enhanced along with an increased MOI and an extended time of transfection .HUVECs and HASMCs transfected with rAAV 9-EGFP-R65 began to express EGFP at 24 h after transfection .The expression peak appeared on the sixth day in HUVECs, and the fifth day in HASMCs.The efficiencies of transfection in HUVECs at MOI of 1×105, 1×106 and 1×107 on the sixth day were (1.40±1.20)%, (12.30±1.35)%and (52.80±2.05)%, respectively.The trans-fection efficiencies of HASMCs on the fifth day were (5.30±1.04)%, (18.30±2.24)% and (52.40±3.21)%at MOI of 1×105 , 1×106 and 1×107 .Cell growth and morphology were not affected by transfection .Alamar Blue assay confirmed that there was no significant difference in the absorbance value between the transfected cells and two types of control cells .Western blot assay showed that the expression of NF-κB P65 was decreased by the trans-fection of rAAV9-EGFP-R65 in HUVECs and HASMCs .Conclusion rAAV9-EGFP-R65 can be efficiently trans-fected into two types of human vascular cells .It shows no inhibitory effects on cell proliferation , but can repress NF-κB P65 expression.

Background::The causal relationship between visceral adipose tissue (VAT) and gout is still unclear. We aimed to examine the potential association between them using observational and Mendelian randomization (MR) analyses.Methods::In the observational analyses, a total of 11,967 participants (aged 39.5 ± 11.5 years) were included from the National Health and Nutrition Examination Survey. Logistic regression models were used to investigate the association between VAT mass and the risk of gout. In two-sample MR analyses, 211 VAT mass-related independent genetic variants (derived from genome-wide association studies in 325,153 UK biobank participants) were used as instrumental variables. The random-effects inverse-variance weighted (IVW) method was used as the primary analysis. Additional sensitivity analyses were also performed to validate our results.Results::Observational analyses found that an increase in VAT mass (per standard deviation) was associated with a higher risk of gout after controlling for confounding factors (odds ratio [OR] = 1.27, 95% confidence intervals [CI] = 1.11–1.45). The two-sample MR analyses demonstrated a causal relationship between increased VAT mass and the risk of gout in primary analyses (OR = 1.78, 95% CI = 1.57–2.03). Sensitivity analyses also showed similar findings, including MR-Egger, weighted median, simple mode, weighted mode, and leave-one-out analyses.Conclusions::Observational analyses showed a robust association of VAT mass with the risk of gout. Meanwhile, MR analyses also provided evidence of a causal relationship between them. In summary, our findings suggested that targeted interventions for VAT mass may be beneficial to prevent gout.

Objective:To investigate the effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation.Methods:This is a retrospective cohort study. The clinical data of 399 patients who underwent orthotopic liver transplantation at the First Affiliated Hospital of Nanjing Medical University from January 2016 to September 2020 were retrospectively collected. There were 319 males and 80 females, aged (50.9±10.2) years (range: 10 to 73 years). These patients were divided into the control group (369 cases) and the dexmedetomidine group (30 cases) according to whether dexmedetomidine was continuously pumped intravenously during the operation until the operation ended. The 1∶2 propensity score matching was used to match the preoperative and intraoperative conditions of the two groups of patients, and the caliper width was 0.2. Outcome indicators included intraoperative postreperfusion syndrome, acute kidney injury and pulmonary complications within 7 days after surgery, length of hospital stay, time of stay in ICU, duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival and recurrence-free survival rate after surgery. The independent sample t test, χ2 test, Mann-Whitney U test or Fisher exact test was used to statistically analyze the data of the two groups of patients, respectively. Survival curves of overall survival and disease-free-survival were plotted by Kaplan-Meier method, and the survival rate and recurrence-free survival rate were compared by Log-rank test. Results:A total of 78 patients were included after propensity score matching, including 26 in the dexmedetomidine group and 52 in the control group. The incidence of acute kidney injury in the dexmedetomidine group within 7 days after surgery was 0 (0/26), significantly lower than that of the control group (21.2%,11/52)(corrected χ2=4.776, P=0.029). There were no significant differences in the incidence of intraoperative postreperfusion syndrome and pulmonary complications within 7 days after surgery, length of hospital stay, ICU time, the duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival, and recurrence-free survival rate after surgery between the two groups (all P>0.05). Conclusion:Continuous infusion of dexmedetomidine via intravenous pump during operation may be beneficial in reducing the incidence of acute kidney injury within 7 days after orthotopic liver transplantation.

Objective:To investigate the effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation.Methods:This is a retrospective cohort study. The clinical data of 399 patients who underwent orthotopic liver transplantation at the First Affiliated Hospital of Nanjing Medical University from January 2016 to September 2020 were retrospectively collected. There were 319 males and 80 females, aged (50.9±10.2) years (range: 10 to 73 years). These patients were divided into the control group (369 cases) and the dexmedetomidine group (30 cases) according to whether dexmedetomidine was continuously pumped intravenously during the operation until the operation ended. The 1∶2 propensity score matching was used to match the preoperative and intraoperative conditions of the two groups of patients, and the caliper width was 0.2. Outcome indicators included intraoperative postreperfusion syndrome, acute kidney injury and pulmonary complications within 7 days after surgery, length of hospital stay, time of stay in ICU, duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival and recurrence-free survival rate after surgery. The independent sample t test, χ2 test, Mann-Whitney U test or Fisher exact test was used to statistically analyze the data of the two groups of patients, respectively. Survival curves of overall survival and disease-free-survival were plotted by Kaplan-Meier method, and the survival rate and recurrence-free survival rate were compared by Log-rank test. Results:A total of 78 patients were included after propensity score matching, including 26 in the dexmedetomidine group and 52 in the control group. The incidence of acute kidney injury in the dexmedetomidine group within 7 days after surgery was 0 (0/26), significantly lower than that of the control group (21.2%,11/52)(corrected χ2=4.776, P=0.029). There were no significant differences in the incidence of intraoperative postreperfusion syndrome and pulmonary complications within 7 days after surgery, length of hospital stay, ICU time, the duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival, and recurrence-free survival rate after surgery between the two groups (all P>0.05). Conclusion:Continuous infusion of dexmedetomidine via intravenous pump during operation may be beneficial in reducing the incidence of acute kidney injury within 7 days after orthotopic liver transplantation.

Objective:To analyze the risk factors and prognosis for early acute kidney injury (AKI) after orthotopic liver transplantation (OLT).Methods:The retrospective study was conduc-ted. The clinicopathological data of 340 pairs of donor and recipients undergoing OLT in The First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. There were 262 males and 78 females of donors. There were 268 males and 72 females of recipients, aged (51±11)years. Of 340 recipients, 217 cases without postoperative early AKI were divided into the non-AKI group and 123 cases with postoperative early AKI were divided into the AKI group. Measure-ment data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distri-bution were represented as M( IQR), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was conducted using the non-parameter test. Multivariate analysis was conducted using the binary Logistic regre-ssion model with forward method. The nomogram predictive model was constructed using the R software with its RMS package (R3.6.1). The efficacy of the predictive model was validated using the area under curve (AUC) of the receiver operating characteristic (ROC) curve, and internal validation of the predictive model was performed using the Bootstrap method. The Kaplan-Meier method was used to draw survival curves, and Log-rank test was used for survival analysis. Results:(1) Com-parison of preoperative clinical characteristics between donors and recipients of the non-AKI group and the AKI group. There was a significant difference in overweight of donors between the non-AKI group and the AKI group ( P<0.05). There were significant differences in preoperative hypertension, viral hepatitis, pathological types, international normalized ratio, fibrinogen levels, platelet (PLT), hemoglobin, and anemia of recipients between the non-AKI group and the AKI group ( P<0.05). (2) Comparison of surgical situations between recipients of the non-AKI group and the AKI group. There were significant differences in intraoperative urine output, volume of intraoperative blood loss, peak serum potassium after reperfusion, massive transfusion, plasma infusion, cryoprecipitate infusion, and aminocaproic acid use of recipients between the non-AKI group and the AKI group ( P<0.05). (3) Influencing factors for postoperative early AKI and construction and evaluation of the nomogram predictive model for postoperative early AKI. Results of multivariate analysis showed that donors of overweight, recipients of preoperative hypertension, recipients of non-viral hepatitis, recipients of preoperative severe PLT reduction, recipients of less intraoperative urine output, recipients of severe post-reperfusion hypotension, recipients of high peak serum potassium after reperfusion, recipients with intraoperative plasma infusion were independent risk factors for postoperative early AKI ( odds ratio=1.982, 3.365, 0.519, 3.615, 0.169, 2.480, 1.500, 1.001, 95% confidence interval as 1.160-3.388, 1.649-6.865, 0.293-0.917, 1.358-9.621, 0.061-0.464, 1.246-4.934, 1.003-2.243, 1.000-1.001, P<0.05). The nomogram predictive model for postoperative early AKI was constructed based on the results of multivariate analysis. Results of ROC curve showed the AUC was 0.769 (95% confidence interval as 0.717-0.820). Results of the calibration curve showed that the predictive results of nomogram predictive model fitted well with the actual situation, with a mean absolute error of 0.016. (4) Comparison of prognosis between recipients of the non-AKI group and the AKI group. There were significant differences in postopera-tive peak creatinine, peak brain natriuretic peptide, duration of intensive care unit stay, mechanical ventilation time, re-intubation of recipients between the non-AKI group and the AKI group ( Z=-4.836, -5.652, -5.861, -6.533, χ2=14.676, P<0.05). All 340 recipients were followed up. For recipients of hepatocellular carcinoma, the 6-month survival rates after surgery were 87.8% and 75.6% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=4.010, P<0.05), and the overall survival rates were 46.7% and 56.1% of the non-AKI group and the AKI group, respectively, showing no significant difference between them ( χ2=0.047, P>0.05). For recipients of benign liver disease, the 6-month survival rates after surgery were 89.8% and 78.0% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=6.401, P<0.05), and the overall survival rates were 81.4% and 68.0% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=4.452, P<0.05). Conclusions:Donors of overweight, recipients of preoperative hypertension, recipients of non-viral hepatitis, recipients of preoperative severe PLT reduction, reci-pients of less intraoperative urine output, recipients of severe post-reperfusion hypotension, recipi-ents of high peak serum potassium after reperfusion, recipients with intraoperative plasma transfu-sion were independent risk factors for postoperative early AKI. Nomogram predictive model has well clinical application value. For recipients of benign liver disease, the 6-month survival rate after surgery and overall survival rate of recipients in the non-AKI group are superior to those of the AKI group.

Eleven compounds were isolated and purified from the ethyl acetate part of 80% ethanol extract of Ferula feruloides root by a combination of normal-phase silica gel column chromatography, Sephadex LH-20 dextran gel column chromatography and semi-preparative liquid chromatography and then modern wave spectrometry methods (NMR, MS, UV, IR) were used to identify the structures of the compounds, which were identified as baigene D (1), baigene E (2), baigene F (3), β-kirialovin (4), α-kirialovin (5), falcarindiol (6), ammoresinol (7), dshamirone (8), 2,3-dihydro-7-hydroxy-2S*,3R*-dimethyl-3-[4-methyl-5-(4-methy1-2-furyl)-3(E)-pentenyl]-furo[3,2-c]coumarin (9), 2,3-dihydro-7-hydroxy-2S*,3R*-dimethyl-2-[4,8-dimethyl-3(E),7-nonadi-enyl]-furo[3,2-c]coumarin (10), and baigene C (11). Compounds 1-3 are new coumarin analogues, and compounds 4-6 are firstly isolated from F. feruloides. The anti-proliferative activity of compounds 5, 7-11 against human gastric cancer (MKN-45) cells was evaluated using MTT assay, which showed that compounds 7-11 exhibited strong inhibitory activity, and compound 5 exhibited weak inhibitory activity.