Aim To investigate the effect of hydrogen sulfide on hepatic lipid accumulation in obese mice. Methods C57 BL/6 J mice were randomly divided into control group, model group, and NaHS group. The mice of the control group were fed with normal diet. The mice of the model group and the NaHS group were fed with high-fat diet. From the thirteenth week, the mice of NaHS group were injected intraperitoneally with NaHS (H2S donor) in a dose of 50 μmol·kg-1 per day for 4 weeks and the mice of the model group were injected with the same volume of saline. All mice were sacrificed at the end of the 16th week. The tis-sues of liver were homogenized and centrifugated. The supernatants were used for the determination of triglyc-eride and cholesterol in liver. The morphology of liver was tested by H&E staining. Liver lipid accumulation was determined by oil red staining. Total RNA was ex-tracted from frozen tissue of liver. PCR was used to de-tect CPT-1 , FAS gene expression and ELISA method was used to detect CPT-1,FAS activity in mice liver. Results The body weight of the mice from NaHS group and model group was bigger than that of the mice from control group. Compared with the model group, the body weight of the mice from NaHS group was less;the content of triglyceride and cholesterol in liver was lower; the degree of liver tissue pathological changes and lipid accumulation were alleviated; CPT-1 expres-sion and activity were increased; FAS expression and activity were decreased. Conclusions These data in-dicate that hydrogen sulfide can reduce the lipid con-tent of liver tissue in obese mice and alleviate fatty liv-er. The mechanism may be associated with the in-creased expression of CPT-1 and the decreased expres-sion of FAS in liver.

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H2S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes.METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro.The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups: the cells in control group were incubated with normal medium for 54 h;the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h;the cells in H2S group or DL-propar-gylglycine (PAG;an inhibitor of cystathione γ-lysase, inhibiting H2S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured.RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H2S group decreased significantly, while those increased significantly in PAG group.CONCLUSION: In steatosis hepatocytes, exogenous H2S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.

Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes and nitrite treatment reduces total triglyceride levels in the high-fat diet mice. Therefore, the present study aimed to determine the effects of nitrite on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1.2 mmol · L(-1) oleic acid (OA) for 24 h. Secondly, steatotic L-02 cells were treated with 0.2 mmol · L(-1) sodium nitrite (SN) plus 3-methyladenine (3-MA), or chloroquine (CQ) for 24 h, and then lipid accumulation was measured with oil red O staining and triglyceride quantification. The notable steatosis could be observed in L-02 cells following exposure to 1.2 mmol · L(-1) OA for 24 h. Treatment with 0.2 mmol · L(-1) sodium nitrite reduced lipid accumulation in steatotic L-02 cells. 3-MA weakened the ability of sodium nitrite to ameliorate hepatic steatosis. Additionally, the sodium nitrite increased number of LC3-II immunostaining puncta and LC3-II protein expression was confirmed by immunofluorescence or Western blot analysis, and the effects were enhanced by CQ treatment. The number of increased cytoplasm vacuoles and lysosomes increased was confirmed by phase contrast and fluorescence microscope respectively. The increased autolysosome was detected by electron microscopy, this phenomenon could be reversed by CQ treatment. These data demonstrated that sodium nitrite enhanced the autophagic flux and decomposition of triglycerides in steatotic L-02 cells.

Objective To investigate the renal cortical perfusion parameters assessed by contrast-enhanced ultrasound(CEUS)in elderly patients with moderate-to severe renal artery stenosis (RAS),and relationship of those with glomerular filtration rate(GFR).Methods A total of 59 elderly patients with unilateral moderate to severe RAS(50％-99％)admitted into Beijing Hospital during June 2017 to December 2018 were consecutively enrolled in the cross sectional and self-control study.Based on the GFR assessed by radionuclide renal dynamic imaging,patients were divided into the mild-(35＞GFR≥25 ml/min,n=18),moderate-(25 ＞ GFR≥ 15 ml/min,n =37) and severe renal dysfunction(GFR＜ 15 ml · min-1,n =4) groups.CEUS evaluated-renal cortical perfusion parameters,including area under curve (AUC),ascending slope (A),derived peak intensity (DPI),time to peak (TTP) and mean transit time(MTT),were compared among different levels of severity of renal dysfunction groups.Pearson correlation analysis was used to evaluate the relationship between CEUS imaging-evaluated renal cortical perfusion parameters and radionuclide renal dynamic imaging-assessed GFR.Results Compared with the mild renal dysfunction group,the moderate renal dysfunction group showed a reduced AUC and A(P＜0.05),and the severe renal dysfunction group showed more markedly reduced AUC,A,DPI,TTP and MTT (P ＜ 0.05).Compared with the moderate renal dysfunction group,the severe renal dysfunction group showed a reduced AUC,A and GFR(P ＜0.05),while no significant differences in TTP and MTT were found.As far as relationship between renal cortical perfusion parameters and GFR were concerned,Pearson correlation analysis demonstrated that AUC(r =0.804),A(r =0.813) and DPI(r =0.733) were significantly and positively correlated with GFR(all P ＜ 0.05).However,renal function (r =-0.721),TTP(r =-0.744) and MTT(r =-0.743)were negatively correlated with GFR(P＜0.05).And the degree of renal artery stenosis was poorly correlated with GFR (r =-0.462).Conclusions For elderly patients with unilateral moderate to severe RAS,contrast-enhanced ultrasonography can display the changes of renal cortical blood perfusion parameters along with the aggravation of renal function impairment.AUC and A are strongly positively correlated with GFR.The degree of renal artery stenosis is poorly correlated with renal function.And more studies are needed to confirm these findings.

Objective: To evaluate the characteristics of renal cortical blood perfusion assessed by contrast-enhanced ultrasound (CEUS) in elderly patients with renal artery stenosis (RAS) and its relationship with renal function. Methods: Ninety-three elderly patients diagnosed with RAS, who were admitted in Beijing Hospital during June 2017 and December 2018, were retrospectively enrolled. According to the degree of RAS, 186 renal arteries were divided into normal renal artery group (n=79), mild RAS group (30% to 49%, n=59), moderate RAS group (50% to 70%, n=33), and severe RAS group (70% to 99%, n=15). Renal cortical blood perfusion and renal glomerular filtration rate (GFR) were measured by CEUS and radionuclide renal dynamic imaging. According to the renal GFR, 186 kidneys were divided into normal renal function group (GFR≥35 ml/min, n=42) and mild renal insufficiency group (35 ml/min>GFR≥25 ml/min, n=51), moderate renal insufficiency group (25 ml/min>GFR≥15 ml/min, n=75) and severe renal insufficiency group (GFR<15 ml/min, n=18). The renal cortical blood perfusion time-intensity curve (TIC) and related parameters were analyzed, including the area under the curve (AUC), the slope of the ascending branch (A), the peak intensity (PI), the peak time (TTP) and the mean transit time (MTT), the kidneys of different RAS groups and patients with different renal function groups were analyzed. Pearson correlation analysis was used to evaluate the correlation between renal cortical blood perfusion parameters and renal GFR. Results: (1) Renal cortical blood perfusion and GFR: CEUS showed that parameter A of TIC was significantly reduced, while TTP was prolonged in the mild renal artery stenosis group compared with the normal renal artery group (both P<0.05), GFP was similar between the two groups. Cortical perfusion parameters, such as AUC, A, PI and GFR were significantly lower, while TTP and MTT were significantly prolonged in the moderate and severe renal artery stenosis group than in the normal and mild stenosis groups (all P<0.05). Compared with the moderate stenosis group, AUC, A, PI and GFR were significantly lower while TTP, MTT were significantly prolonged in the severe renal artery stenosis group (all P<0.05). (2) TIC showed that the renal perfusion parameters, AUC, PI and A were significantly lower, while TTP was significantly longer in the mild renal dysfunction group than in the normal renal function group (all P<0.001). The changes aggravated in proportion with renal dysfunction. (3) Correlation between perfusion parameters and GFR: Pearson correlation analysis showed that the AUC (r=0.774, P<0.05), A (r=0.815, P<0.05) and PI (r=0.772, P<0.05) were positively correlated with GFR; serum creatinine level (r=-0.841, P<0.05), renal function grading (r=-0.731, P<0.05), TTP (r=-0.803, P<0.05) and MTT (r=-0.741, P<0.05) were negative correlated with GFR. The degree of stenosis was negatively correlated with GFR (r=-0.427, P<0.05). Conclusion Cortical perfusion parameters differ significantly among patients with various degree of RAS and renal dysfunction. The renal cortical blood perfusion parameters are correlated with renal GFR.

Recently introduced microhaplotype has attracted more attention in forensic field. A microhaplotype is a short segment of DNA (e.g<200bp) contains two or more SNPs. The microhaplotypes showed extremely low mutation rates. The amplicons of alleles are balance to each other and without any stutters. By these advantages, it shows great value for forensic application, such as mixture stains analyzing, ancestor analysis, and kinship analysis. Thus, we aimed to review the development, analytical approach, nomenclature and population characteristic of microhaplotype.

Objective:To explore the potential role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/GATA-binding protein 4 (GATA-4)/vascular endothelial growth factor (VEGF) signal pathway in neovascular age-related macular degeneration (nAMD).Methods:We applied the TRANSFAC Public database to search the human and mouse VEGF promoters and upstream transcription factors, analyzed the transcription factors that may influence the transcriptional activity of VEGF. The RAW264.7 cells were divided into control group and lipopolysaccharide (LPS) stimulated group (LPS group). Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the activation of NLRP3 inflammasome, and the mRNA levels of GATA-4 and VEGFA. Thus, we applied the specific small molecular NLRP3 inhibitor MCC950 pretreated RAW264.7 cells (LPS+ MCC950 group), and detected the gene expression of NLRP3, Caspase-1, interleukin 1β( IL-1β), GATA-4 and VEGFA.Results:There were multiple GATA transcription factor binding sites upstream of human and mouse VEGF promoters. Compared with the control group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS group were increased (all P<0.05). Compared with LPS group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS+ MCC950 group were significantly decreased (all P<0.05). Conclusions:NLRP3/GATA-4/VEGF signal pathway may play a significant role in the pathologic processes of nAMD.

Objective To investigate the ameliorative effect of salidroside on diabetes retinopathy (DR) rats and its mechanism. Methods Male SD rats were randomly divided into blank group, model group, low-dose and high-dose salidroside treatment groups. Except for the blank group, other groups were modeled by intraperitoneal injection of streptozotocin. After successful modeling, treatment groups were injected intraperitoneally with [50 mg/(kg.d)] and [100 mg/(kg.d)] salidroside respectively, for 4 weeks; the blank group and model group were injected with corresponding doses of saline. ELISA was used to measure the expression levels of antioxidant-related enzyme activity and inflammatory factors in blood glucose and serum of rats in each group. Retinal tissue lesions were detected by HE staining, and the expression of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule 1 (ICAM-1) in retinal tissues were detected by immunohistochemical staining. Western blot analysis was used to detect the expression of phosphatidylinositol 3 kinase (PI3K) , nuclear factor κB p65 (NF-κB p65), phosphorylated p38 MAPK (p-p38 MAPK), and phosphorylated protein kinase B (p-AKT) proteins. Results Compared with model group, salidroside could significantly reduce blood glucose level and increase body mass in DR rats. The serum levels of superoxide dismutase (SOD) and catalase (CAT) were significantly increased, while the levels of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and IL-1β were reduced. The protein expression of VEGF, ICAM-1, NF-κB p65 and p-p38 MAPK was significantly decreased, while the protein expression of PI3K and p-AKT was increased. Conclusion Salidroside can reduce DR in rats by inhibiting oxidative stress and immune inflammatory response, which may be related to the reduction of abnormal expression of VEGF and ICAM-1 and the activation of PI3K/AKT signaling pathway.
Animals ; Male ; Rats ; Blood Glucose ; Diabetes Mellitus ; Inflammation/metabolism* ; Intercellular Adhesion Molecule-1/metabolism* ; NF-kappa B/metabolism* ; Oxidative Stress ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Rats, Sprague-Dawley ; Retinal Diseases ; Tumor Necrosis Factor-alpha/metabolism* ; Vascular Endothelial Growth Factor A/metabolism*

Objective:To investigate the imaging characteristics on contrast-enhanced ultrasound(CEUS)of carotid plaque instability and their correlation with histopathological changes in elderly patients.Methods:Twenty-three patients undergone carotid endarterectomy(CEA)at the Department of Neurosurgery of Beijing Hospital from June 2018 to November 2020 were retrospectively enrolled, including 27 males and 5 females, aging from 60 to 87(68.1±6.3)years.They were diagnosed with moderate to severe carotid artery stenosis(50%-99%)based on digital subtraction angiography(DSA).Carotid artery CEUS was conducted before surgery, and HE staining and CD34 immunohistochemical staining were used to record histopathological scores and microvessel density(MVD)counts.Results:The mean pathological score and MVD count were 2.46±0.66 and 37.17±12.88 for 32 cases with hypoechoic patterns, 22.42±0.55 and 38.06±13.66 for 18 cases with mixed echo patterns, and 2.75±0.35 and 23.50±9.192 for 2 cases with strong echo patterns, respectively.CEUS grading was positively correlated with histopathology score and MVD count( r=0.53、0.76, all P<0.01). Conclusions:Low echo and mixed echo plaques are more unstable than strong echo plaques.CEUS can be used to comprehensively assess the vulnerability of atherosclerotic plaques and to decide the optimal surgical time for patients with carotid atherosclerotic plaque stenosis.

Objective:To establish and validate a prognostic model of a contrast-enhanced ultrasound scoring(CEUS)system for evaluating renal artery stenosis(RAS)in the elderly.Methods:This was a single-center retrospective study.A total of 324 elderly RAS patients admitted to Beijing Hospital from October 2017 to July 2020 were randomly assigned into the model group(n=174)and the validation group(150)in a 1∶1 ratio.Clinical and imaging data of patients on admission including general conditions, previous medical history, blood pressure, blood creatinine, renal artery stenosis and cortical blood perfusion in the affected kidney and renal function(GFR)at 1-year follow-up were collected.Univariate and multivariate logistic regression was used to establish a model of the CEUS scoring system.The receiver operating characteristic(ROC)curve and area under the ROC curve(AUC)were used to evaluate prediction accuracy.Clinical application value of the CEUS scoring system model was evaluated via decision curve analysis using a nomogram.Results:Baseline clinical and radiomic data had no significant difference between the model group and the validation group( P>0.05). Multivariate logistic regression analysis results showed that age( OR=1.242, 95% CI: 1.081-1.427, P<0.01), diabetes( OR=1.545, 95% CI: 1.107-2.156, P<0.05), blood pressure( OR=1.328, 95% CI: 1.056-1.670, P<0.05), renal function( OR=2.374, 95% CI: 1.216-3.887, P<0.01)and cortical blood perfusion parameter( OR=2.646, 95% CI: 1.553-6.369, P<0.01)were risk factors for the deterioration of renal function during 1 year follow-up.Based on these results, a nomogram for the CEUS scoring system model was drawn, and its consistency index, the C-Index, was 0.725(95% CI: 0.653-0.776). The AUC of the CEUS scoring system was 0.824 and the Youden index was 0.711 in the model group, with a specificity of 0.774 and a sensitivity of 0.837.The AUC of the CEUS scoring system was 0.853 and the Youden index was 0.715 in the validation group, with a specificity of 0.684 and a sensitivity of 0.889.There was no significant difference in ROC curve between the two groups( D=1.387, P>0.05). In addition, calibration charts of the two models showed that the calibration curve of the CEUS scoring system was close to the standard curve, with no statistically significant difference( P>0.05). Conclusions:The CEUS scoring system model can be used to predict the risk of worsening renal function in elderly RAS patients during 1-year follow-up.