Therapeutic mechanism of emodin for treatment of rheumatoid arthritis: a network pharmacology-based analysis / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic mechanism of emodin in the treatment of rheumatoid arthritis (RA) using a network pharmacology-based method and validate this mechanism in a fibroblast-like synovial cell line.@*METHODS@#The PubChem, Targetnet, SwissTargetPrediction, Genecards, OMIM, and DisGeNET databases were searched to obtain emodin targets and RA-related genes. A protein-protein interaction (PPI) network was constructed, and GO and KEGG pathway enrichment analyses were carried out to analyze the intersection genes. AutoDock4.2.6 software was used to simulate molecular docking between emodin and its candidate targets. In a cultured fibroblast-like synovial cell line (MH7A), the effects of different concentrations of emodin on proliferation of tumor necrosis factor-α (TNF-α)-induced cells were investigated using CCK-8 assay, cell scratch experiment and flow cytometry; the changes in the expressions of nuclear factor-κB (NF-κB) pathway proteins were detected using Western blotting, and the mRNA expressions of the hub genes were examined with RT-qPCR.@*RESULTS@#We identified 32 intersection genes of emodin and RA, and the key targets including CAPS3, ESR1, and MAPK14 involved mainly the NF-κB signaling pathway. Cell scratch experiment and flow cytometry demonstrated a strong inhibitory effect of emodin on MH7A cell proliferation. Treatment with TNF-α significantly increased the cellular expressions of the NF-κB pathway proteins, which were obviously lowered by treatment with 80 μmol/L emodin. The results of RT-qPCR showed that TNF-α treatment obviously up-regulated the expressions of the hub genes COX2 and P38MAPK, and emodin treatment significantly down-regulated the expressions of MAPK and PTGS2 and up-regulated the expression of CASP3.@*CONCLUSION@#The therapeutic effect of emodin on RA is mediated mainly through regulation of cell proliferation, apoptosis, and the NF-κB pathway.

Protective Effect of Modified Danshenyin and Erchentang on Intimal Injury Caused by Carotid Atherosclerosis / 中国实验方剂学杂志

Objective:To discuss the clinical efficacy of modified Danshenyin and Erchentang in treating carotid atherosclerosis （CAS）， and the effect on intimal injury. Method:Patients （151 cases） were divided into control group （75 cases） and observation group （76 cases）. Specifically， 69 cases in control finished the treatment （4 cases fell off in follow-up， and 2 cases were eliminated）， and 69 cases in observation group finished the treatment （3 cases fell off in follow-up， and 4 cases were eliminated）. Patients in both group got atorvastatin calcium tablets， 10 mg/time， 1 time/day， and aspirin enteric-coated tablets， 100 mg/time， 1 time/day. Patients in control group got Hedan tablets， 2 tablets/time， 3 times/day. Patients in observation group got modified Danshenyin and Erchentang， 1 dose/day. The treatment lasted for 4 months. Before and after treatment， color Doppler ultrasound of carotid artery was detected， and carotid intima-media thickness （IMT）， plaque number， plaque area， plaque thickness and hemodynamics were recorded. Levels of nitric oxide （NO）， endothelin-1 （ET-1）， von Willebrand factor （vWF）， soluble intercellular adhesion molecule-1 （sICAM-1）， vascular endothelial growth factor （VEGF）， matrix metalloproteinase-9 （MMP-9）， triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， whole-blood low-shear viscosity （LBV）， whole-blood high-shear viscosity （HBV）， plasma viscosity （PV）， platelet aggregation rate （PAR）， fibrinogen （FIB）， homocysteine （Hcy）， interleukin-6 （IL-6）， IL-10， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， serum superoxide dismutase （SOD）， malondialdehyde （MDA）， oxidized low density lipoprotein （ox LDL） and circulating glutathione peroxidase （GSH-Px） were detected before and after treatment. And the safety was evaluated. Result:After treatment， IMT， number， area and thickness of plaque in observation group were less than those in control group （<italic>P</italic><0.01）. Peak systolic velocity and end diastolic velocity in observation group were higher than those in control group （<italic>P</italic><0.01）， while pulsatility index and resistance index were lower than those in control group （<italic>P</italic><0.01）. And levels of ET-1， vWF， sICAM-1， VEGF， MMP-9， TG， TC， LDL-C， LBV， HBV， PV， PAR， FIB， Hcy， IL-6， TNF-<italic>α</italic>， MDA and ox-LDL were lower than those in control group （<italic>P</italic><0.01）， whereas levels of NO， HDL-C， IL-10， SOD and GSH-Px were higher than those in control group （<italic>P</italic><0.01）. And there was no adverse reaction caused by traditional Chinese medicine. Conclusion:Modified Danshenyin and Erchentang can reduce plaque， improve hemodynamics and hemorheology， and regulate blood lipid metabolism and vascular endothelial factor， with anti-inflammatory and anti-oxidation damages. It can protect vascular intima， and inhibit the occurrence and development of CAS， with a safety in clinical use.

Effects of Wuqinxi on Balance, Walking and Quality of Life for Patients with Parkinson's Disease / 中国康复理论与实践

Objective:To observe the effects of traditional Chinese exercise Wuqinxi on balance, walking and quality of life for patients with Parkinson's disease. Methods:From December, 2018 to December, 2020, 62 inpatients with Parkinson's disease in the Affiliated Hospital of Shandong University of Chinese Medicine were randomly divided into control group (n = 31) and observation group (n = 31). The control group received routine medicine and rehabilitation training, while the observation group received Wuqinxi in addition, for eight weeks. The trajectory length and ellipse area of the center of pressure in 30 seconds were measured with PRO-KIN before and after treatment, while they were assessed with Berg Balance Scale (BBS), Timed 'Up and Go' Test (TUGT) and Parkinson's Disease Quality of Life Scale-39 (PDQ-39). Results:The trajectory length and ellipse area of the center of pressure, BBS score, TUGT time, and PDQ-39 score improved in both groups after treatment (t > 11.225, P < 0.001), and all improved more in the observation group than in the control group (t > 5.919, P < 0.001). Conclusions:Wuqinxi training is effective on balance, walking and quality of life for patients with Parkinson's disease.

Therapeutic silencing miR-146b-5p improves cardiac remodeling in a porcine model of myocardial infarction by modulating the wound reparative phenotype

Fibrotic remodeling is an adverse consequence of immune response-driven phenotypic modulation of cardiac cells following myocardial infarction (MI). MicroRNA-146b (miR-146b) is an active regulator of immunomodulation, but its function in the cardiac inflammatory cascade and its clinical implication in fibrotic remodeling following MI remain largely unknown. Herein, miR-146b-5p was found to be upregulated in the infarcted myocardium of mice and the serum of myocardial ischemia patients. Gain- and loss-of-function experiments demonstrated that miR-146b-5p was a hypoxia-induced regulator that governed the pro-fibrotic phenotype transition of cardiac cells. Overexpression of miR-146b-5p activated fibroblast proliferation, migration, and fibroblast-to-myofibroblast transition, impaired endothelial cell function and stress survival, and disturbed macrophage paracrine signaling. Interestingly, the opposite effects were observed when miR-146b-5p expression was inhibited. Luciferase assays and rescue studies demonstrated that the miR-146b-5p target genes mediating the above phenotypic modulations included interleukin 1 receptor associated kinase 1 (IRAK1) and carcinoembryonic antigen related cell adhesion molecule 1 (CEACAM1). Local delivery of a miR-146b-5p antagomir significantly reduced fibrosis and cell death, and upregulated capillary and reparative macrophages in the infarcted myocardium to restore cardiac remodeling and function in both mouse and porcine MI models. Local inhibition of miR-146b-5p may represent a novel therapeutic approach to treat cardiac fibrotic remodeling and dysfunction following MI.

Role of aloperine on ischemia-reperfusion-induced injury and inflamma-tory response in H9c2 cardiomyocytes / 中国病理生理杂志

AIM:To explore the role of aloperine in ischemia-reperfusion(I/R)-induced H9c2 cardiomyocyte injury and inflammation.METHODS: The H9c2 cardiomyocytes were cultured under hypoxia and re-oxygenation condi-tions to simulate ischemia-reperfusion(SI/R)injury.After treatment with aloperine at various doses,the cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.Simultaneously,the levels of lactate dehydro-genase(LDH),malonaldehyde(MDA)and caspase-3 activity were detected by the commercial kits.The levels of inflam-matory cytokines were also detected by ELISA.Moreover,the effects of aloperine on the activation of PI 3K/AKT signaling pathway were determined by Western blot.RESULTS:Pre-treatment with aloperine remarkably abated the inhibitory effect of SI/R on H9c2 cell viability,and decreased the elevations of LDH and MDA triggered by SI /R(P<0.05).Pre-treat-ment with aloperine dramatically suppressed the cell apoptosis induced by SI /R treatment(P<0.05), concomitant with the decrease in caspase-3 activity and increase in Bcl-2/Bax ratio(P<0.05).In contrast to SI/R group,aloperine treat-ment notably restrained the concentrations of pro-inflammatory cytokines, including interleukin-6, tumor necrosis factor-α and interleukin-1β(P<0.05).Furthermore, aloperine remarkably increased the protein levels of p-PI3K and p-AKT. While blocking the PI3K/AKT pathway with its specific inhibitor LY294002, the viability-promoting, anti-apoptotic and anti-inflammatory effects of aloperine on the H 9c2 cells were obviously attenuated(P<0.05).CONCLUSION: Alope-rine protects against cardiomyocytes from I/R injury and inhibits inflammatory responses by activating the PI 3K/AKT signa-ling pathway,implying a potential benefic role of aloperine against myocardial I /R injury.

An improved method for preparation and evaluation of a porcine model of acute myocardial infarction by minimally invasive surgery and left circumflex artery ligation / 中国实验动物学报

Objective To develop a better method for preparation of porcine model of acute myocardial infarction by permanent occlusion of the left circumflex coronary artery and minimally invasive surgery, evaluate its validity and stability, and explore its application in experimental studies of ischemic heart diseases.Methods 25 healthy female 3-month-old Bama minipigs, body weight 25±3 kg, were used in this study.The porcine model of myocardial infarction was established by minimally invasive surgery and the left circumflex artery ligation at the site of OM1 posterior position under general anesthesia.Heart function was assessed by echocardiography at 15 min before surgery, 1 hour and 4 weeks after surgery.Pathological examination was performed at 4 weeks after the left circumflex artery occlusion.The mortality and cause of death were statistically analyzed.Results The 1-hour and 4-week postoperative cardiac function was considerably decreased, showing a decreased ejection fraction from 64.2±4.6% to 48.2±5.3% (1hour after MI) and 49.7±6.1% (4 weeks after MI) (P<0.01).Pathological examination revealed that the ventricular wall was thinner and the amount of collagens was increased in the infracted area.The ventricular fibrillation rate at 1-hour after myocardial infarction was 17.3% and the infarction area was 19.2%.Conclusions A pig model of acute myocardial infarction can be prepared by our modified left circumflex coronary artery ligation at the obtuse marginal artery (OM1) and minimally invasive surgery.This model exhibits advantages such as minimal surgical trauma, high stability of the model, and low mortality, therefore, provides an ideal and economic animal model for experimental studies on acute ischemic heart diseases.

Breeding and microbiological quality control of germ-free pigs / 中国实验动物学报

The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.

A porcine model of orthotopic left lung transplantation / 中国实验动物学报

Objective To establish a porcine allogeneic left lung orthotopic transplantation model to closely simu-late human lung transplantation.Methods Twelve Huanjiang mini-pigs were used as donors and 12 Bama mini-pigs as recipients.The left lung orthotopic transplantation was completed by the left fourth intercostal thoracotomy.At 1 h, 2 h, 4 h, 6 h, 12 h after transplantation, the left and right pulmonary artery pressure were measured, the left and right pulmonary vein blood gas was analyzed, and samples of the left and right lung tissues were taken to determine the water content and for pathological examination.Results All animals survived, and the transplanted pulmonary vein blood PaO2/FiO2 and PAP were rised along with the prolonged postoperative time, compared with those of the recipient normal lung showing a signifi-cant difference (P<0.05).With the pass of time, there were increasing edema, inflammatory cell infiltration, RBC ooze, thickening of alveolar wall in the transplanted lung tissue, and some alveolar lumen occlusion and lung tissue consolidation. The water content of the transplanted lung tissue was increased significantly compared with that in the recipient lung tissue ( P<0.05 ) .Conclusions The established method in this study provides an ideal animal model for research on lung transplantation ischemia-reperfusion injury and immune rejection mechanism.

Experience and improvement in establishing an abdominal working heart transplantation model in rats / 中国实验动物学报

Objective To modify the techniques for establishment of an abdominal working heart transplantation model in rats and to sum up the key factors to success.Methods A total of 180 12-week old Brown Norway rats ( donor) and Lewis rats ( recipient) were used in this study:50 BN rats and 50 Lewis rats for pilot experiment, and 40 BN rats and 50 Lewis rats for the formal experiment.The rat model of working heart heterotopic transplantation was adopted and estab-lished by Wiedemann’ s mode.We transplanted the heart from BN rats to Lewis rats and analyzed the survival rate, causes of death and histological changes of the heart ( HE staining) in this experiment.Results After exercise and modification, the survival rate was increased to 77.5%, and the mean total duration of operation was 71 ±11 min, and the mean ische-mic time of the donor hearts was 34 ±5 min.Histological examination ( HE staining) of the cardiac allograft showed a mild inflammatory cell infiltration in the graft at 24 h after transplantation, indicating that the model was reliable.Conclusions A variety of factors may affect the final operation success rate in the establishment of this heart transplantation model.A-mong them, the major affecting factors include: healthy animals, donor heart protection, rapid and effective vascular su-ture, and postoperative animal management.

Establishment of a sheep model of pulsatile ventricular assist device / 中国实验动物学报

Objective To establish a large animal ( sheep) model to serve the experiments of domestic pulsatile ventricular assist device.Methods Three small-tail Han-sheep were anesthetized and the vein access and artery access were achieved.The cardiopulmonary bypass was established through left thoracotomy.Ventricular fibrillation was induced. An hole was made in the apex of left ventricle and the apex cannulation was sutured to it.The aortic cannulation was su-tured to the descending aorta.The two cannulations were connected to the domestic pulsatile ventricular assist device ( DP-VAD) and the driver was turned on.The working of DPVAD and the conditions of the animals were observed.Results The DPVAD worked well and uni-directional blood flow was driven by positive and negative pressure.The left ventricle was unloaded and the blood pressure was raised up.Conclusion The establishment of sheep model of pulsatile ventrieular as-sist device may play important role for the research and development of DPVAD in our country.

Differential expression profiles of microRNAs in a rat model of obliterative bronchiolitis / 中国组织工程研究

BACKGROUND:There is no effective therapy for obliterative bronchiolitis after tracheal transplantation. A therapeutic strategy at microRNA (miRNA) molecular level plays a crucial role in the prevention and treatment of complications after organ transplantation. OBJECTIVE:To analyze the miRNA differential expression profile in response to obliterative bronchiolitis after orthotopic tracheal transplantation in rats. METHODS:The obliterative bronchiolitis model after lung transplantation was established through orthotopic tracheal transplantation in inbred strains of rats, and then was identified using histoIogical examination. Total miRNAs were detected by miRNA array and significantly differential expressed miRNAs were filtrated in the transplanted trachea tissues. The miRNA-146a, miRNA-155 and miRNA-451 with significantly differential expressions were used for relative quantitative study. Quantitative real-time reverse transcription-polymerase chain reaction was applied to verify the reliability of miRNA array results. RESULTS AND CONCLUSION:The pathological examination showed that, obliterative bronchiolitis model in rats was successful y established at 4 weeks after orthotopic tracheal transplantation. A total of obliterative bronchiolitis-related 29 miRNAs were found in miRNA expression profiles, including 14 miRNAs with significantly down-regulated expression and 15 miRNAs with significantly up-regulated expression. Among them, the significantly up-regulated miRNAs (miRNA-146a and miRNA-155) and the significantly down-regulated miRNA-451 were involved in immuno-inflammatory reaction. The miRNAs play an important role in regulating pathophysiological changes of obliterative bronchiolitis after lung transplantation.

Assessment of Bax gene transfer influence on survival of heterotopic cardiac allograft in rats by high-frequency echocardiography / 中华超声影像学杂志

Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P ＜ 0.05] and group A [(7.26 ± 1.50)d,P ＜0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P ＜0.05) while no significant difference was found between group B and C(P ＞0.05).At day 3 and 6,LVT of group A and B were higher than group C (P ＜0.05) while LVTR was lower(P ＜0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.

Interleukin 17 decrease the polarization of macrophage and attenuate the pathology of obliterative bronchiolitis after trachea transplant modulated / 中华胸心血管外科杂志

Objective To investigate the role of interleukin 17 in the differentiation of macrophages from monocyte in vivo and its applicability in Obliterative bronchiolitis (OB).Methods Pathogen-free male C57BL/6 (n =20),IL-17-defcient C57BL/6 (n =10) and BALB/c (n =10) weighing 20-25 g.Weight-matched mice were assigned to three experimental groups.Experimental group A:BALB/c (n =5) →C57 BL/6 (n =10).Experimental group B:BALB/c (n =5) →IL-17-defcient C57BL/6(n =10).Normal control group:C57BL/6(n =10).The CD80 expression in macrophages in spleens were analysed by flowcytometry 7 days after operation.Native and transplanted lungs were harvested after 14 days and 28 days,stained with HE,and examined under light microscopy.Results Compared to experimental group A,CD80 expression of macrophages reveales a significant decrease in spleen of experimental group B.Furthermore,airway obliteration and destruction of the epithelium in experimental group A were significantly better compared with that of experimental group B on day 14 and 28 after transplantation.Conclusion IL-17 deficiency decreased the polarization of monocytes to macrophages and attenuate the pathology of obliteral bronchiolitis in murine model after trachea transplantaion.

Dual-source CT in the diagnosis of tetralogy of Fallot / 中华胸心血管外科杂志

Objective To investegate the value of dual-source CT (DsCT) in the diagnosis of tetralogy of Fallot (TOF).Methods The measurement results of the main pulmonary artery( PA),left pulmonary artery (LPA),right pulmonary artery( RPA ),right entricular outflow tract(ROVT) of 42 cases with TOF by DsCT,echocardiography aud surgery were ret rospectively analyzed.Results Of the 42 cases,a total of 232 cardiac nomalies were found by surgery ( 145 intra cardiac anomalies,53 ventricular-aterial connection anomalies,34 external cardiac anomalies),DsCT and echocardiography found 204 cardiac nomalies (130 intra cardiac anomalies,12 ventricular-aterial connection anomalies,53 external cardiac anomalies),224 cardiac nomalies ( 145 intra cardiac anomalies,31 ventricular-aterial connection anomalies,48 external cardiac anomalies)respectively;and the diagnostic accuracy of DsCT and echocardiography was 88％ (204/232),97％ (224/232) respectively.There was not significant difference between DsCT and surgery ( P ＞ 0.05 ).There was significant difference between echocardiography and surgery in PA,LPA and RPA (P＜0.05),but not in ROVT and over-riding of the aorta (P＞0.05).There was not significant difference between DsCT and echocardiography ( P ＞ 0.05 ).Conclusion Echocardiography was superior to DsCT in intra cardiac anomalies,especially in the cardiac septal defects and heart valve diseases.DsCT had the advantages in external cardiac anomalies,particularly in assessing pulmonary artery.

The effects of CD28 superagonist antibody in rat obliterative airway disease / 中国免疫学杂志

Objective:To investigate the effects of superagonistic CD28~- specific monoclonal antibody JJ316 (supCD28 MAb) on preferentially expanded rat CD4~+CD25~+Treg (Treg) cells in vivo and its applicability in obliterative airway disease (OAD).Methods:The heterotopic tracheal transplantation model in rats was used.One group received mIgG-treatment(0.5 mg/rat) as control.The experimental group was treated with supCD28 Mab(0.5 mg/rat) via intraperitoneal injection on the day of transplantation.The changes of Treg cell population in cervical lymph nodes were monitored by flow cytometry after 7 days.Tracheas were harvested after 21 days for further histologic evaluation.Results:SupCD28 MAb administration was revealed with a significant increase in the CD4~+CD25~+ T and CD4~+Foxp3~+ T cells population in cervical lymph nodes compared to treatment with mIgG group on day 7 after transplantation,[8.5%±3.4% and 11.5%±2.7% (P<0.05 vs mIgG group)in the supCD28 Mab group,1.8%±1.9% and 3.2%±2.1% in the mIgG group,respectively].Furthermore,the allografts from animals treated with supCD28 MAb were significantly less airway obliteration and destruction of the epithelium compared with that of control group animals on day 21 after transplantation.Conclusion:SupCD28 MAb targets expansion of Treg cells and attenuates airway lumen obliteration in rat obliterative airway disease.

Updated biological characterization of Th17 cells / 医学研究生学报

Th17 cells play an important role in the pathogenesis of various immune-mediated diseases,including psoriasis,rheumatoid arthritis,multiple sclerosis,inflammatory bowel disease,asthma,and so on.The discovery of Th17 cells has offered scientists a new insight into the etiology and treatment of a broad spectrum of diseases.This article presents an overview of the differentiation,cytokine expression and trafficking of Th17 cells.