OBJECTIVE: To identify the role of gene silencing or overexpression of Nemo-like kinase (NLK) during the process of neural differentiation of human mesenchymal stem cells (hBMSCs), and to explore the effect of NLK downregulation by transfection of small interfering RNA (siRNA) on promoting neuralized tissue engineered bone regeneration. METHODS: NLK-knockdown hBMSCs were established by transfection of siRNA (the experimental group was transfected with siRNA silencing the NLK gene, the control group was transfected with control siRNA and labeled as negative control group), and NLK-overexpression hBMSCs were established using lentivirus vector transfection technique (the experimental group was infected with lentivirus overexpressing the NLK gene, the control group was infected with an empty vector lentivirus and labeled as the empty vector group). After neurogenic induction, quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression of neural-related gene, and Western blot as well as immunofluorescence staining about several specific neural markers were used to evaluate the neural differentiation ability of hBMSCs.6-week-old male nude mice were divided into 4 groups: ① β-tricalcium phosphate (β-TCP) group, ② β-TCP+ osteogenic induced hBMSCs group, ③ β-TCP+ siRNA-negative control (siRNA-NC) transfection hBMSCs group, ④ β-TCP+ siRNA-NLK transfection hBMSCs group. Four weeks after the subcutaneous ectopic osteogenesis models were established, the osteogenesis and neurogenesis were detected by hematoxylin-eosin (HE) staining, Masson staining and tissue immunofluorescence assay. Statistical analysis was conducted by independent sample t test. RESULTS: After gene silencing of NLK by siRNA in hBMSCs, neural-related genes, including the class Ⅲ β-tubulin (TUBB3), microtubule association protein-2 (MAP2), soluble protein-100 (S100), nestin (NES), NG2 proteoglycan (NG2) and calcitonin gene-related peptide (CGRP), were increased significantly in NLK-knockdown hBMSCs compared with the negative control group(P < 0.05), and the expression levels of TUBB3 and MAP2 of the NLK silencing group were also increased. Oppositely, after NLK was overexpressed using lentivirus vector transfection technique, TUBB3, MAP2, S100 and NG2 were significantly decreased in NLK-overexpression hBMSCs compared with the empty vector group (P < 0.05), and the expression level of TUBB3 was also decreased. 4 weeks after the subcutaneous ectopic osteogenesis model was established, more mineralized tissues were formed in the β-TCP+ siRNA-NLK transfection hBMSCs group compared with the other three groups, and the expression of BMP2 and S100 was higher in the β-TCP+ siRNA-NLK transfection hBMSCs group than in the other groups. CONCLUSION Gene silencing of NLK by siRNA promoted the ability of neural differentiation of hBMSCs in vitro and promoted neuralized tissue engineered bone formation in subcutaneous ectopic osteogenic models in vivo in nude mice.
Bone Regeneration/genetics* ; Animals ; Mesenchymal Stem Cells/cytology* ; Humans ; RNA, Small Interfering/genetics* ; Tissue Engineering/methods* ; Cell Differentiation ; Mice, Nude ; Gene Silencing ; Mice ; Male ; Protein Serine-Threonine Kinases/genetics* ; Intracellular Signaling Peptides and Proteins/genetics* ; Transfection ; Cells, Cultured ; Lentivirus/genetics*

Objectives:To investigate the clinical value of cardiac magnetic resonance imaging(CMR)feature-tracking strain analysis in risk stratification of diabetic heart failure with preserved ejection fraction(HFpEF).Methods:In this retrospective study,a total of 215 patients with diabetic HFpEF who underwent CMR at Chinese Academy of Medical Sciences Fuwai Hospital from January 2012 to December 2018 were included.Myocardial strain parameters were calculated using CMR feature-tracking technology.Patients were followed up by medical records or telephone calls.Composite endpoint event,all-cause death or heart failure hospitalization during follow-up were recorded.Patients were divided into event group and event-free group.Univariable and multivariable Cox proportional hazard regression analyses were performed to determine the risk factors for the outcomes in diabetic HFpEF.The effects of hypertension and obesity on the prognosis of diabetic HFpEF patients and whether they affect the prognostic value of CMR feature-tracking strain analysis were also analyzed.Results:During a follow-up of(7.1±1.8)years,93(43.3%)patients had endpoint events(event group),including 28 all-cause deaths and 65 heart failure hospitalization.Compared with the event-free group(n=122),patients in the event group had significantly lower left ventricular ejection fraction,higher prevalence and extent of late gadolinium enhancement,and significantly reduced global longitudinal strain(GLS),global circumferential strain,global radial strain,and global systolic longitudinal strain rate(all P<0.05).The absolute GLS value was significantly lower in event group than in event-free group,regardless of the presence of hypertension and obesity.Multivariate Cox regression analysis showed that estimated glomerular filtration rate(HR=0.983,95%CI:0.972-0.993,P=0.001),left atrial volume index(HR=1.015,95%CI:1.005-1.026,P=0.004),and GLS(HR=1.142,95%CI:1.060-1.231,P<0.001)were independent risk factors for adverse cardiovascular events in diabetic HFpEF patients.However,adjusted N-terminal pro-brain natriuretic peptide was not an independent prognostic factor.The cut-offvalue of GLS to predict outcome was-14.09%from ROC curve analysis.The Kaplan-Meier curve showed that in patients with and without hypertension and obesity,patients with the GLS>-14.09%had lower event-free survival compared to patients with GLS≤-14.09%(all P<0.05),and the ability of GLS to predict adverse outcomes was not affected by hypertension and obesity.Conclusions:GLS obtained by CMR feature-tracking strain analysis is an independent predictor of adverse outcomes in diabetic HFpEF,and its ability to predict adverse outcomes is independent of hypertension and obesity.

Biofilm is one of the important reasons for bacterial resistance and persistent infection in Staphylococcus aureus,which poses a significant threat to the dairy farming industry in southern Xinjiang.Although quercetin has antibacterial properties,its poor hydrophilicity and low solubility limit its clinical application.Therefore,quercetin nanogel was prepared by the electrostatic interac-tion between guar gum(positive charge)and hyaluronic acid(negative charge)under the action of sodium tripolyphosphate in this study.Using encapsulation efficiency and loading capacity as indi-cators,the optimal formula was screened through single factor experiments and response surface methodology,and characterized.The minimal inhibitory concentration(MIC)and minimal mem-brane inhibitory concentration(MBIC)of quercetin nanogel against Staphylococcus aureus were determined by micro broth dilution method and micro plate semi quantitative method.The results showed that the optimal formulation of quercetin nanogel was 11.6 g/L guar gum,10.0 g/L hyalu-ronic acid and 50.0 g/L sodium tripolyphosphate.The encapsulation efficiency and loading capacity were 79.4％and 7.6％,respectively.The particle size and Zeta potential were(396.0±4.2)nm and(-25.0±0.8)mV,respectively,indicating that the nanogel had good dispersion,high stability and excellent drug loading capacity.The MIC and inhibitory rate of quercetin nanogel against Staphy-lococcus aureus were 8.0 mg/L and 83.8％,respectively.The MBIC and inhibitory rate of quercetin nanogel against Staphylococcus aureus were 8.0 mg/L and 38.3％,respectively.Therefore,the pre-pared quercetin nanogel solved the problems of low solubility and poor hydrophilicity of quercetin,improved the inhibitory effect of quercetin on the membrane of Staphylococcus aureus,and was expected to be further applied to the treatment of cow bacterial diseases in southern Xinjiang.

Biofilm is one of the important reasons for bacterial resistance and persistent infection in Staphylococcus aureus,which poses a significant threat to the dairy farming industry in southern Xinjiang.Although quercetin has antibacterial properties,its poor hydrophilicity and low solubility limit its clinical application.Therefore,quercetin nanogel was prepared by the electrostatic interac-tion between guar gum(positive charge)and hyaluronic acid(negative charge)under the action of sodium tripolyphosphate in this study.Using encapsulation efficiency and loading capacity as indi-cators,the optimal formula was screened through single factor experiments and response surface methodology,and characterized.The minimal inhibitory concentration(MIC)and minimal mem-brane inhibitory concentration(MBIC)of quercetin nanogel against Staphylococcus aureus were determined by micro broth dilution method and micro plate semi quantitative method.The results showed that the optimal formulation of quercetin nanogel was 11.6 g/L guar gum,10.0 g/L hyalu-ronic acid and 50.0 g/L sodium tripolyphosphate.The encapsulation efficiency and loading capacity were 79.4％and 7.6％,respectively.The particle size and Zeta potential were(396.0±4.2)nm and(-25.0±0.8)mV,respectively,indicating that the nanogel had good dispersion,high stability and excellent drug loading capacity.The MIC and inhibitory rate of quercetin nanogel against Staphy-lococcus aureus were 8.0 mg/L and 83.8％,respectively.The MBIC and inhibitory rate of quercetin nanogel against Staphylococcus aureus were 8.0 mg/L and 38.3％,respectively.Therefore,the pre-pared quercetin nanogel solved the problems of low solubility and poor hydrophilicity of quercetin,improved the inhibitory effect of quercetin on the membrane of Staphylococcus aureus,and was expected to be further applied to the treatment of cow bacterial diseases in southern Xinjiang.

Objectives:To investigate the clinical value of cardiac magnetic resonance imaging(CMR)feature-tracking strain analysis in risk stratification of diabetic heart failure with preserved ejection fraction(HFpEF).Methods:In this retrospective study,a total of 215 patients with diabetic HFpEF who underwent CMR at Chinese Academy of Medical Sciences Fuwai Hospital from January 2012 to December 2018 were included.Myocardial strain parameters were calculated using CMR feature-tracking technology.Patients were followed up by medical records or telephone calls.Composite endpoint event,all-cause death or heart failure hospitalization during follow-up were recorded.Patients were divided into event group and event-free group.Univariable and multivariable Cox proportional hazard regression analyses were performed to determine the risk factors for the outcomes in diabetic HFpEF.The effects of hypertension and obesity on the prognosis of diabetic HFpEF patients and whether they affect the prognostic value of CMR feature-tracking strain analysis were also analyzed.Results:During a follow-up of(7.1±1.8)years,93(43.3%)patients had endpoint events(event group),including 28 all-cause deaths and 65 heart failure hospitalization.Compared with the event-free group(n=122),patients in the event group had significantly lower left ventricular ejection fraction,higher prevalence and extent of late gadolinium enhancement,and significantly reduced global longitudinal strain(GLS),global circumferential strain,global radial strain,and global systolic longitudinal strain rate(all P<0.05).The absolute GLS value was significantly lower in event group than in event-free group,regardless of the presence of hypertension and obesity.Multivariate Cox regression analysis showed that estimated glomerular filtration rate(HR=0.983,95%CI:0.972-0.993,P=0.001),left atrial volume index(HR=1.015,95%CI:1.005-1.026,P=0.004),and GLS(HR=1.142,95%CI:1.060-1.231,P<0.001)were independent risk factors for adverse cardiovascular events in diabetic HFpEF patients.However,adjusted N-terminal pro-brain natriuretic peptide was not an independent prognostic factor.The cut-offvalue of GLS to predict outcome was-14.09%from ROC curve analysis.The Kaplan-Meier curve showed that in patients with and without hypertension and obesity,patients with the GLS>-14.09%had lower event-free survival compared to patients with GLS≤-14.09%(all P<0.05),and the ability of GLS to predict adverse outcomes was not affected by hypertension and obesity.Conclusions:GLS obtained by CMR feature-tracking strain analysis is an independent predictor of adverse outcomes in diabetic HFpEF,and its ability to predict adverse outcomes is independent of hypertension and obesity.

Objectives:To analyze the consistency of evaluating left ventricular hemodynamics (HDF) based on single plane and multi plane cine sequences of magnetic resonance mitral valve orifice.Methods:A prospective study was conducted on 48 healthy adults, and two methods were used to measure the mitral valve diameter and calculate HDF parameters. The first method was to measure the diameter of the mitral valve opening in the left ventricular three chamber cine sequence; The second method is to measure the mitral valve diameter using cine sequences of two chamber, three chamber, and four chamber hearts, and then take the average value. Paired t-tests were used to compare the differences in HDF measured by two methods, and Pearson correlation coefficient ( r), intra group correlation coefficient ( ICC), and Bland-Altman analysis were used to test the consistency and reproducibility of the two methods. Results:The root mean square (RMS) of longitudinal HDF calculated using single plane and multi plane mitral valve diameters were [(17.28±4.41)% vs (17.21±4.61)%] ( P=0.379) for the entire cardiac cycle, [(21.45±5.54)% vs (21.49±5.68)%] ( P=0.646) for systolic phase, and [(12.78±4.10)% vs (12.54±4.24)%] ( P=0.106) for diastolic phase, respectively. The difference in the calculation results of HDF parameters related to ventricular function was not statistically significant (all P>0.05), and there was good consistency ( r=0.924-0.996, ICC=0.924-0.995). The two HDF parameters related to atrial function were sensitive to the measurement method of mitral valve orifice diameter [RMS of longitudinal HDF during active atrial emptying: (3.26±1.51)% vs (3.32±1.55)%, P=0.006; longitudinal HDF pulse during active atrial emptying: (-2.60±1.28)% vs (-2.76±1.30)%, P<0.001]. Conclusions:The ventricular function related HDF parameters obtained from the analysis of mitral valve orifice diameter using single plane and multi plane methods have good consistency, and can be evaluated using relatively simple single plane methods for left ventricular HDF.

The assessment of cardiac function using echocardiography has gained a strong foothold in clinical practice. Cardiac magnetic resonance (CMR) imaging harbors distinct advantages over echocardiography, as it is not affected by limitations of acoustic windows and operator dependence. CMR is also designed to non-invasively assess cardiac morphology, ventricular geometry, myocardial wall motion, and intra-cardiac flow quantification without the use of ionizing radiation. These inherent features make CMR appropriate for diagnosing cardiovascular diseases, monitoring patients after treatment, and providing longitudinal follow-up. In this paper, the state-of-the-art work that has demonstrated the aspects of cardiac function by CMR is reviewed, and acquisition techniques and clinical applications are covered.

The assessment of cardiac function using echocardiography has gained a strong foothold in clinical practice. Cardiac magnetic resonance (CMR) imaging harbors distinct advantages over echocardiography, as it is not affected by limitations of acoustic windows and operator dependence. CMR is also designed to non-invasively assess cardiac morphology, ventricular geometry, myocardial wall motion, and intra-cardiac flow quantification without the use of ionizing radiation. These inherent features make CMR appropriate for diagnosing cardiovascular diseases, monitoring patients after treatment, and providing longitudinal follow-up. In this paper, the state-of-the-art work that has demonstrated the aspects of cardiac function by CMR is reviewed, and acquisition techniques and clinical applications are covered.

OBJECTIVE To investigated the anti-de-pressant effects of the fruit Areca catechu L.(ACL)and elucidated its potential underlying mechanism using a rat model of chronic unpredictable mild stress(CUMS).METHODS CUMS was induced in rats to establish a depression animal model for 28 d.According to the baseline sucrose preference,the male rats were divided into six different groups.They were treated with parox-etine hydrochloride,ACL,and water once a day until the behavioral tests were performed.The levels of corticoste-rone(CORT),malondialdehyde(MDA),catalase(CAT),and total superoxide dismutase(T-SOD)in serum were de-tected using a commercial kit,and the concentrations of 5-hydroxytryptamine(5-HT)and dopamine(DA)mono-amine neurotransmitters in the brain tissues were detect-ed by liquid chromatography-tandem mass spectrometry.Doublecortin(DCX)expression in the hippocampal den-tate gyrus(DG)was determined by immunofluorescence,and the relative abundance of brain-derived neurotrophic factor(BDNF),TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β of brain tissues were assayed by West-ern blotting.RESULTS ACL markedly increased sucrose preference,decreased the immobility time,and short-ened the feeding latency of CUMS-induced rats.CUMS induction resulted in marked changes in the contents of the monoamine neurotransmitters(5-HT and DA)in the hippocampus and cortex of brain tissues and the levels of CORT,MDA,CAT,and T-SOD in serum,whereas ACL administration alleviated these considerable changes.ACL promoted DCX expression in DG and increased the protein levels of BDNF,TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β in the brains of CUMS-induced rats.CONCLUSION Our results indicated that ACL may improve depression-like behaviors in CUMS-induced rats by decreasing the hyperfunction and oxidative stress of the hypothalamic-pituitary-adrenal axis,stimulating hippo-campal neurogenesis,and activating the BDNF signaling pathway.

Objective:To investigate the effect of galactose lectin 3 (Gal-3) on the pathogenesis of atrial fibrillation.Methods:This study adopts a case-control study method. 55 patients with non valvular atrial fibrillation (atrial fibrillation group) admitted to the First People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from February to July 2019 were selected, and another 55 healthy individuals who underwent physical examination at our hospital during the same period were selected as the control group. Compare the general data and levels of various laboratory indicators between two groups, including blood routine, fasting blood glucose, blood lipids, liver and kidney function, and plasma Gal-3. Analyze the influencing factors of atrial fibrillation and the predictive value of plasma Gal-3 levels for the onset of atrial fibrillation. The measurement data with normal distribution and the measurement data converted to normal distribution after taking natural logarithm are expressed in xˉ± s. The comparison between the two groups is performed by independent sample t test; The measurement data of non normal distribution is represented by [ M ( Q1, Q3)], and Wilcoxon signed rank sum test is used for inter group comparison; The counting data is represented by examples (%), and the comparison between groups is conducted using χ 2 test. The influencing factors of atrial fibrillation were analyzed using logistic regression analysis. Results:The age, NLR, and blood creatinine levels in the atrial fibrillation group were higher than those in the control group [(71.16±9.17) years vs (60.71±10.11) years, (2.32±0.85) vs (1.74±0.81), (74.18±21.61) μmol/L vs (64.69±18.30) μmol/L, t-values are 5.68, 3.66, 2.48, P-values are <0.001, <0.001, 0.015], total cholesterol, HDL-C, LDL-C Albumin and eGFR water were on average lower than those in the control group [(4.31±1.67) mmol/L vs (5.13±0.78) mmol/L, (0.96±0.21) mmol/L vs (1.21±0.32) mmol/L, (2.35±0.65) mmol/L vs (3.04±0.62) mmol/L, (39.58±3.83) g/L vs (44.66±5.61) g/L, (94.84±29.22) mL/(min·1.73 m 2) vs (111.77±21.51) mL/(min·1.73 m 2)] ,The t-values are 3.30, 4.87, 5.69, 5.54, 3.46, and the P-values are 0.001,<0.001,<0.001,<0.001, 0.001, respectively. The plasma Gal-3 levels in the atrial fibrillation group were higher than those in the control group [(12.79±4.24)] μg/L vs (7.31±2.28) μg/L], the difference was statistically significant ( t=8.43, P<0.001), and the plasma Gal-3 level in the persistent atrial fibrillation group was higher than that in the paroxysmal atrial fibrillation group [(14.03±3.95) μg/L vs (11.51±4.21) μg/L], the difference was statistically significant ( t=2.29, P=0.026). The results of multivariate logistic regression analysis showed that after excluding other factors, Gal-3 remained an independent influencing factor for atrial fibrillation (odds ratio=1.66, 95% confidence interval: 1.29-2.12, P<0.001). Conclusions:Plasma Gal-3 is an influencing factor for the onset of atrial fibrillation. After excluding other factors, Gal-3 remains an independent influencing factor for atrial fibrillation, with an increase of 1 μg/L in Gal-3 increases the risk of atrial fibrillation by 1.66 times.