ObjectiveTo investigate the therapeutic effects and mechanisms of Qinlian Hongqutang （QLHQT） on nonalcoholic steatohepatitis （NASH）. MethodsC57BL/6J mice were randomly divided into normal and modeling groups. The NASH model was established by feeding a high-fat diet for 12 weeks. After successful modeling， mice were randomly assigned to the model group， low-， medium-， and high-dose QLHQT groups （0.51， 1.02， and 2.04 g·kg^-1）， and a positive control metformin group， with six mice in each group. The mice were treated for 8 weeks. Body weight was recorded before and after treatment. Serum levels of total cholesterol （TC）， triglycerides （TG）， and low-density lipoprotein cholesterol （LDL-C）， as well as hepatic TC， TG， and LDL-C contents， were determined by biochemical assays. Hematoxylin-eosin （HE） staining and oil red O staining were used to evaluate liver histopathology and lipid deposition， respectively. Flow cytometry， enzyme-linked immunosorbent assay （ELISA）， and Real-time polymerase chain reaction （Real-time PCR） were used to assess hepatic macrophage expression and related markers. Western blot and immunofluorescence were used to investigate the potential mechanisms of QLHQT in regulating macrophage polarization. ResultsCompared with the normal group， body weight and serum and hepatic levels of TC， TG， and LDL-C were significantly increased in the model group （P<0.01）. Liver histopathology showed unevenly distributed round lipid droplets in the hepatocyte cytoplasm， accompanied by inflammatory cell aggregation. Flow cytometry showed that the proportion of CD86-positive cells was significantly increased， whereas the proportion of CD206-positive cells was markedly decreased （P<0.05）. Hepatic inducible nitric oxide synthase （iNOS） levels and tumor necrosis factor-α （TNF-α） mRNA expression were significantly increased， while hepatic IL-10 levels and IL-4 mRNA expression were significantly decreased （P<0.01）. The protein expression levels of Toll-like receptor 4 （TLR4）， tumor necrosis factor receptor-associated factor 6 （TRAF6）， and myeloid differentiation factor 88 （MyD88） in the liver were significantly increased （P<0.01）. Compared with the model group， body weight was reduced in the high-， medium-， and low-dose QLHQT groups and in the metformin group. Serum and hepatic TC， TG， and LDL-C levels were significantly decreased （P<0.01）. Liver histopathology showed alleviated hepatic lipid deposition， with markedly reduced lipid droplets and inflammation. Immunofluorescence and flow cytometry showed that the proportions of CD86-positive cells were significantly decreased， whereas the proportions of CD206-positive cells were significantly increased in the high-， medium-， and low-dose QLHQT groups （P<0.05）. Hepatic iNOS levels and TNF-α mRNA expression were significantly decreased （P<0.01）， whereas hepatic IL-10 levels and IL-4 mRNA expression were significantly increased （P<0.01）. The hepatic protein expression levels of TLR4， TRAF6， and MyD88 were significantly decreased， while signal transducer and activator of transcription 6 （STAT6） phosphorylation was significantly increased （P<0.05， P<0.01）. There was no statistically significant difference in total STAT6 protein expression. ConclusionQLHQT effectively ameliorates hepatic inflammation in NASH mice， and the mechanism may involve STAT6- and TLR4-mediated signaling pathways driving polarization of M1 macrophages toward the M2 phenotype.

ObjectiveTo investigate the therapeutic effects and mechanisms of Qinlian Hongqutang （QLHQT） on nonalcoholic steatohepatitis （NASH）. MethodsC57BL/6J mice were randomly divided into normal and modeling groups. The NASH model was established by feeding a high-fat diet for 12 weeks. After successful modeling， mice were randomly assigned to the model group， low-， medium-， and high-dose QLHQT groups （0.51， 1.02， and 2.04 g·kg^-1）， and a positive control metformin group， with six mice in each group. The mice were treated for 8 weeks. Body weight was recorded before and after treatment. Serum levels of total cholesterol （TC）， triglycerides （TG）， and low-density lipoprotein cholesterol （LDL-C）， as well as hepatic TC， TG， and LDL-C contents， were determined by biochemical assays. Hematoxylin-eosin （HE） staining and oil red O staining were used to evaluate liver histopathology and lipid deposition， respectively. Flow cytometry， enzyme-linked immunosorbent assay （ELISA）， and Real-time polymerase chain reaction （Real-time PCR） were used to assess hepatic macrophage expression and related markers. Western blot and immunofluorescence were used to investigate the potential mechanisms of QLHQT in regulating macrophage polarization. ResultsCompared with the normal group， body weight and serum and hepatic levels of TC， TG， and LDL-C were significantly increased in the model group （P<0.01）. Liver histopathology showed unevenly distributed round lipid droplets in the hepatocyte cytoplasm， accompanied by inflammatory cell aggregation. Flow cytometry showed that the proportion of CD86-positive cells was significantly increased， whereas the proportion of CD206-positive cells was markedly decreased （P<0.05）. Hepatic inducible nitric oxide synthase （iNOS） levels and tumor necrosis factor-α （TNF-α） mRNA expression were significantly increased， while hepatic IL-10 levels and IL-4 mRNA expression were significantly decreased （P<0.01）. The protein expression levels of Toll-like receptor 4 （TLR4）， tumor necrosis factor receptor-associated factor 6 （TRAF6）， and myeloid differentiation factor 88 （MyD88） in the liver were significantly increased （P<0.01）. Compared with the model group， body weight was reduced in the high-， medium-， and low-dose QLHQT groups and in the metformin group. Serum and hepatic TC， TG， and LDL-C levels were significantly decreased （P<0.01）. Liver histopathology showed alleviated hepatic lipid deposition， with markedly reduced lipid droplets and inflammation. Immunofluorescence and flow cytometry showed that the proportions of CD86-positive cells were significantly decreased， whereas the proportions of CD206-positive cells were significantly increased in the high-， medium-， and low-dose QLHQT groups （P<0.05）. Hepatic iNOS levels and TNF-α mRNA expression were significantly decreased （P<0.01）， whereas hepatic IL-10 levels and IL-4 mRNA expression were significantly increased （P<0.01）. The hepatic protein expression levels of TLR4， TRAF6， and MyD88 were significantly decreased， while signal transducer and activator of transcription 6 （STAT6） phosphorylation was significantly increased （P<0.05， P<0.01）. There was no statistically significant difference in total STAT6 protein expression. ConclusionQLHQT effectively ameliorates hepatic inflammation in NASH mice， and the mechanism may involve STAT6- and TLR4-mediated signaling pathways driving polarization of M1 macrophages toward the M2 phenotype.

Objective    To assess outcomes of transcatheter aortic valve replacement (TAVR) for pure native aortic valve regurgitation. Methods    A total of 129 patients underwent transfemoral TAVR in Fuwai Hospital from May 2019 to October 2020 were retrospectively analyzed. There were 83 males and 46 females with an average age of 72.26±8.97 years. The patients were divided into a pure native aortic valve regurgitation group (17 patients) and an aortic valve stenosis group (112 patients). Results    The incidence of valve in valve was higher in the pure native aortic valve regurgitation group (47.0% vs. 16.1%, P<0.01). There was no statistical difference between the two groups in conversion to surgery, intraoperative use of extracorporeal circulation, intraoperative left ventricular rupture, postoperative use of extracorporeal membrane oxygenation (ECMO), peripheral vascular complications, disabled stroke, death, or pacemaker implantation. There was no statistical difference in the diameter of annulus (25.75±2.21 mm vs. 24.70±2.90 mm, P=0.068) or diameter of outflow tract (25.82±3.75 mm vs. 25.37±3.92 mm, P=0.514) between the pure native aortic valve regurgitation group and the aortic valve stenosis group. Conclusion    Transfemoral TAVR is a feasible method for patients with pure native aortic valve regurgitation. The diameter of annulus plane, the diameter of outflow tract and the shape of outflow tract should be evaluated.

Objective:To detect the expression differences of Wnt signaling pathway related molecules β-catenin, Cyclin D1 and Dickkopf-1 (DKK1) in breast disease tissues, and to explore their application value in pathologically aided diagnosis of breast cancer.Methods:From January 2008 to August 2019, 90 cases of breast tissue specimens in the Cancer Center of Guangzhou Medical University were collected, including 30 cases of breast hyperplasia, 30 cases of breast intraductal carcinoma and 30 cases of breast invasive ductal carcinoma. The expressions of β-catenin, Cyclin D1 and DKK1 in breast tissue of each group were detected by immunohistochemistry. Oncomine database and KM plotter database were used to analyze the expression differences of β-catenin, Cyclin D1 and DKK1 in breast cancer and normal breast tissues and their relationships with survival prognosis of patients with breast cancer, and to verify the results of immunohistochemistry. Receiver operating characteristic (ROC) curve was used to evaluate the efficacies of each molecule in pathologically aided diagnosis.Results:There were statistically significant differences in β-catenin, Cyclin D1 and DKK1 expressions among breast hyperplasia, breast intraductal carcinoma and breast invasive ductal carcinoma ( χ2=7.766, P=0.021; χ2=24.133, P<0.001; χ2=11.585, P=0.003). The expression of β-catenin in breast invasive ductal carcinoma group was significantly higher than that in breast intraductal carcinoma group and breast hyperplasia group ( Z=-2.367, P=0.018; Z=-2.462, P=0.014). The expression of Cyclin D1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-4.166, P<0.001; Z=-4.174, P<0.001). The expression of DKK1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-3.090, P=0.002; Z=-2.923, P=0.003). The results of bioinformatics analysis showed that compared with normal breast tissue, the expression of β-catenin mRNA in invasive breast cancer tissue increased by 2.33 times ( t=15.242, P<0.001), the expression of Cyclin D1 mRNA in breast intraductal carcinoma tissue increased by 6.64 times ( t=7.152, P=0.006), while the expression of DKK1 mRNA in normal breat tissue was 3.41 times higher than that in invasive breast cancer tissue, with no statistically significant difference ( t=-13.193, P>0.999). The median survival time of breast cancer patients in Cyclin D1 high expression group was 173.2 months, which was shorter than 228.9 months in low expression group ( P<0.001). The upper quartile survival time of breast cancer patients in DKK1 high expression group was 55.1 months, which was longer than 40.4 months in low expression group ( P<0.001). The breast invasive ductal carcinoma and breast intraductal carcinoma were combined into tumor group, the sum of the immunohistochemistry scores of β-catenin and Cyclin D1 minus the immunohistochemistry score of DKK1 was used as the combined scoring scheme 1, and the sum of β-catenin and Cyclin D1 immunohistochemistry score was used as the combined scoring scheme 2. ROC curve analysis showed that the area under the curve (AUC) of β-catenin, Cyclin D1, combined scoring scheme 1 and combined scoring scheme 2 for pathologically aided diagnosis of breast cancer were 0.65 ( P=0.080), 0.81 ( P<0.001), 0.70 ( P=0.023) and 0.78 ( P=0.001), respectively. The AUC of Cyclin D1 and combined scoring scheme 2 were ≥0.7, which had good value in pathologically aided diagnosis. Conclusion:Wnt signaling pathway related molecules Cyclin D1 and Cyclin D1 combined with β-catenin detection has a good value in the pathologically aided diagnosis of breast cancer.

Exploratory experiment is the key to the experimental physiology,which is universally applicable training of innovative talents.More importantly,it is an important way to cultivate innovative talents of medicine.On the basis of the original multi-subject evaluation system,we have formed a comprehensive evaluation system through refining the scoring item for teachers and students,enriching the forms of student evaluation,and building the student self-assessment.This comprehensive evaluation system is set to promote the development of experimental physiology,especially the exploratory experiments.The practice of this evaluation system effectively improve the objectivity of the evaluation,provide a basis for reform of exploratory experiment,and then promote the training of innovative talents.

Inflammation plays an important role in occurrence and progression of atherosclerosis.Among numerous studies on inflammatory markers, the relationship between high sensitive C-reactive protein (hsCRP) and coronary heart disease (CHD) has been widely studied.The present article made a review about the relationship between hsCRP and CHD.

Objective:To observe influence of enhanced external counterpulsation (EECP) on plasma levels of lipo‐protein-associated phospholipase A2 (Lp‐PLA2) and hsCRP in patients with coronary heart disease (CHD) ,and preliminarily explore anti -inflammatory mechanism of EECP in prevention and treatment of CHD .Methods :A to‐tal of 85 CHD patients were selected from our hospital ,randomly divided into routine treatment group (n=42) and EECP group [n=43 ,received EECP based on routine treatment ,once/d ,60min each time ,continuous five weeks were regarded as one course (35 times)] .Plasma concentrations of Lp‐PLA2 and hsCRP were measured in all sub‐jects at the enrollment day and after five -week treatment .Results:Compared with before treatment ,after five-week treatment ,there were significant reductions in plasma levels of Lp‐PLA2 [(58.46 ± 40.04)μg/L vs .(33.94 ± 23.22)μg/L] and hsCRP [ (3.54 ± 2.22)μg/ml vs .(2.19 ± 1.16)μg/ml] in EECP group (P<0.01 both) ,and there were no significant difference in routine treatment group between before and after treatment .Compared with routine treatment group ,after five-week treatment ,there were significant reductions in plasma levels of Lp‐PLA2 [ (56.87 ± 33.69)μg/L vs .(33.94 ± 23.22)μg/L] and hsCRP [ (3.63 ± 1.60)μg/ml vs .(2.19 ± 1.16)μg/ml] in EECP group , P<0.01 both .Conclusion:1. EECP of one course can significantly reduce plasma concentrations of Lp‐PLA2 and hsCRP in CHD patients , indicating that anti - inflammatory may be one of its mechanism in preven‐tion and treatment of CHD .

[ ABSTRACT] AIM:To compare the differences of the genome-wide methylation levels and methylated regions be-tween nasopharyngeal carcinoma ( NPC) cells in the same genetic background but different radiation resistance ( CNE-2 cells and CNE-2R cells).METHODS:Using the method which was developed by Doctor Zhao Cun-you, based on using methyl-sensitive restriction enzyme to measure the genome-wide methylation levels.In addition, MeDIP-Seq was used to analyze the methylated regions in 6 gene functional elements, including the upstream 2k sequence, 5’ UTR, coding se-quence, intron, 3’UTR and downstream 2k sequence, between CNE-2 cells and CNE-2R cells.RESULTS:The genome-wide methylation level was approximately 30%lower in CNE-2R cells than that in CNE-2 cells.No obvious difference on the amount of genes and the coverage of the peak in the 6 gene functional elements was observed.However, the methylation pattern of plentiful genes had altered in the gene function elements.CONCLUSION:The genome-wide methylation levels and methylated regions between NPC cells in the same genetic background but different radiation resistance were quite dif-ferent, indicating that the DNA methylation may be associated with NPC radioresistance.

Objective:To explore the relationship among plasma level of lipoprotein‐associated phospholipase A2 (Lp‐PLA2) ,ankle brachial index (ABI) ,brachial‐ankle pulse wave velocity (baPWV) and coronary heart disease (CHD) .Methods :According to results of coronary angiography ,a total of 120 patients with suspected or diagnosed CHD were divided into CHD group (n=90) and non‐CHD group (n=30) .CHD group was further divided into sin‐gle vessel coronary disease group (single vessel group ,n=30) ,double‐vessel coronary disease group (double‐vessel group ,n=30) and multi‐vessel coronary disease group (multi‐vessel group ,n= 30) .Plasma Lp‐PLA2 level ,ABI and baPWV were measured and compared among all groups .Results:Compared with non‐CHD group ,there were significant rise in plasma Lp‐PLA2 level [ (23.60 ± 13.33)μg/L vs .(36.65 ± 17.24)μg/L] and baPWV [ (1244.27 ± 127.85) cm/s vs .(1753.08 ± 284.32) cm/s] in CHD group ,P<0.01 both .In CHD group ,compared with single vessel group ,plasma Lp‐PLA2 level significantly rose [ (25.81 ± 8.97)μg/L vs .(35.03 ± 9.80)μg/L vs .(49.13 ± 21.22)μg/L] in double‐vessel group and multi‐vessel group ,and that of multi‐vessel group was significantly higher than that of double‐vessel group ( P<0.05 or <0.01 );baPWV significantly rose [ (1579.77 ± 178.05 ) cm/s vs . (1808.07 ± 272.11) cm/s ,(1871.40 ± 306.03) cm/s] in double‐vessel group and multi‐vessel group , P<0.01 both . ABI of single vessel group and double‐vessel group were significantly higher than that of multi‐vessle group [ (1.19 ± 0.08) ,(1.17 ± 0.07) vs .(1.11 ± 0.15)] ,P<0.01 or <0.05 .Conclusion:Plasma Lp‐PLA2 level ,ABI and baP‐WV are related to CHD and its lesion degree ,combined measurement of these three indexes can predict CHD and its severity more accurately ,then preventing and treating CHD should be more effective .

Objective To prepare a specific integrin αvβ3 probe 18F-Al-1,4,7-triacetic acid-1,4,7-triazacyclononane-2-(4-amino benzyl) thioamide-(3,6,9-trioxaundecanoic acid-11-amide)-(glutamic acid-cyclo(arginine-glycine-aspartic acid-phenylalanine-lysine) dimeric peptide) (18 F-Al-NOTA-PRGD2) and evaluate its feasibility for PET imaging in papillary thyroid carcinoma (PTC).Methods 18F-Al-NOTA-PRGD2 was synthesized by a novel Al18F complex strategy.Human PTC tissues were implanted into nude mice.Immunohistochemistry staining was performed to detect αvβ3 expression in human PTC tissues,xenografts in mice and adjacent normal tissues respectively.18F-Al-NOTA-PRGD2 with or without blocking agent of PRGD2 was injected via the tail vein into tumor-bearing mice (n =5) for microPET imaging.The radioactivity uptake in the tumor and major organs were measured via ROI technology.Biodistribution studies were also performed in tumor bearing mice (n=15) 30,60,120 min postinjection respectively.The two sample t test was used for statistical analysis.Results The labeling yield of 18F-Al-NOTA-PRGD2 was over 45％ (no attenuation correction) and the radiochemical purity was above 95％.The integrin αvβ3 expression was observed in human PTC both in situ specimens and xenograft in mice,while no expression was shown in the adjacent normal tissues.MicroPET imaging revealed that tumors were clearly visible with good tumor-to-background contrast.The radioactive uptake by tumor was (2.81 ±0.35) ％ ID/g,(2.45±0.27) ％ID/g and (1.80±0.21) ％ID/g at 30,60 and 120 min postinjection,respectively.In the presence of unradiolabeled PRGD2,the corresponding tumor uptake decreased to (0.51±0.05) ％ID/g at 60 min postinjection.High tumor uptake was also shown in the biodistribution studies,which was (3.09±0.25) ％ID/g,(2.75±0.37) ％ID/g and (1.90±0.16) ％ID/g at 30,60 and 120 min postinjection,respectively.The results were consistent with the microPET imaging results (t=1.456,1.465 and 0.847,respectively,all P＞0.0.5).18F-Al-NOTA-PRGD2 was rapidly cleared in blood and muscles,and the tumor to blood and muscle uptake ratios were 6.15±0.45 and 7.86±0.56 respectively.Conclusions 18 F-Al-NOTA-PRGD2 could be labeled easily and quickly with good labeling yield and radiochemical purity.Overexpressed integrin αvβ3 in PTC can be proved by both immunostaining and microPET imaging.18F-A1-NOTA-PRGD2 PET imaging might be a novel in vivo method for investigation of molecular mechanism in PTC.