Mechanism of 3-arylcoumarin derivatives inhibiting vascular calcification / 中国药科大学学报

@#To reveal the pharmacological mechanism of 3-arylcoumarin derivative 3-(4′-hydroxyphenyl)-6-hydroxycoumarin (SJ-6) against vascular calcification, advanced glycation end products (AGEs) were used to induce the calcification of human aortic vascular smooth muscle cells (HCASMCs), and calcification was identified by alizarin red staining and quantification.The effects of SJ-6 on alkaline phosphatase (ALP) activity, cell proliferation rate, calcium content, and total reactive oxygen species (ROS), superoxide dismutase (SOD), AGEs, and tetra methylethlene diamine proteinase factor-α (TNF-α), interleukin-6 (1L-6), interleukin-β (1L-β), runt-related transcription factor 2 mRNA (Runx2 mRNA), the receptor of advanced glycation endproducts (RAGE), nuclear factor kappa-B (NF-κB), napdh oxidase-1 (NoX-1), protein kinase C(PKC), protein kinase b(AKT), p38 mitogen-activated protein kinase (p38 MAPK), and smooth muscle actin-α (SMA-α) protein expression were determined.According to our results, SJ-6 significantly decreased AGEs content, ALP activity, intracellular calcium content, ROS content, Runx2 mRNA and inflammatory factors TNF-α, 1L-6 and 1L-β (P < 0.05) and increased SOD content (P < 0.01), with similar to those of the positive control drug aminoguanidine hydrochloride (AGH).Therefore, we investigated the pharmacological mechanism of compound SJ-6, which was found to significantly inhibit the expression of RAGE, NF-κB, NoX-1, PKC, Akt, p-p38 and other essential signaling proteins in the calcified cell model (P < 0.01) and increas the expression of smooth actin SMA-α (P < 0.01).SJ-6 inhibits vascular calcification by inhibiting oxidative stress and the expression of AGEs/RAGE, Akt/PKC and NF-κB signaling pathways, suggesting that it may be a novel drug for the treatment of vascular calcification.

Effect of targeted silencing Notch1 on proliferation and apoptosis of human non-small cell lung cancer stem cells / 国际肿瘤学杂志

Objective To investigate the effect of targeted silencing Notch1 on proliferation and apoptosis of human non-small cell lung cancer stem cells.Methods Lung cancer A549 cells and SPC-A-1 cells were selected and divided into control group,Nc-shRNA group and Notch1-shRNA group.The Nc-shRNA group was a negative control RNAi lentivirus group,and the Notch1-shRNA group was a Notch1 inhibitory RNAi lentivirus group.The lentiviral-mediated shRNA interference technology was used to target the silencing of Notch1.The silencing effect of Notch1 gene was verified by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting.Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) and sarcosphere formation assay.Apoptosis was detected by Annexin V/7-AAD double staining.Western blotting was used to detect the expression of proliferating cell nuclear antigen (PCNA),B-cell lymphoma-2 (Bcl-2) and Notch1 downstream gene Hes-1.Results The results of qRT-PCR showed that the relative expression levels of Notch1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1 cells were 1.000 ± 0.000,0.937 ± 0.025,0.490 ± 0.036 and 1.000 ± 0.000,1.077 ± 0.070,0.373± 0.038,with statistically significant differences (F =359.707,P ＜0.001;F =210.455,P ＜0.001),further paired comparison,the relative expression of Notch1 in Notch1-shRNA group was significantly lower than that in Nc-shRNA group (all P ＜ 0.05).Western blotting showed that the expressions of Notch1 protein in A549 cells and SPC-A-1 cells were consistent with the mRNA results.MTT assay showed that the 24 h A values of A549 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.209 ± 0.005,0.219 ± 0.009,0.159 ±0.006,48 h A values were 0.293 ± 0.004,0.302 ± 0.004,0.205 ± 0.005,72 h A values were 0.450 ± 0.003,0.430 ± 0.012,0.348 ± 0.017,with statistically significant differences (F =79.487,P＜0.001;F =508.664,P ＜0.001;F =57.156,P ＜0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 24,48,72 h (all P ＜ 0.05).The 48 h A values of SPC-A-1 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.438 ±0.022,0.412 ± 0.015,0.364 ± 0.010,72h A values were 0.540 ± 0.016,0.519 ± 0.009,0.438 ± 0.019,with statistically significant differences (F =15.667,P =0.004;F =37.299,P ＜ 0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 48 h and 72 h (all P ＜ 0.05).The sphere sizes of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were (149.667 ± 6.506) μm,(136.667 ± 7.095) μm,(86.676 ± 7.638) μm,with statistically significant difference (F =65.940,P ＜ 0.001).The sphere sizes of the three groups in SPC-A-1 cells were (118.667 ± 6.658) μm,(128.000 ± 7.000) μm,(60.675 ± 4.509) μm,with statistically significant difference (F =105.372,P ＜0.001).Further paired comparison,the sphere size of Notch1shRNA group was significandy smaller than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).The apoptosis rates of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1cells were (0.489 ± 0.014)％,(0.633 ± 0.021)％,(1.683 ± 0.221)％ and (1.323 ± 0.194)％,(1.690 ± 0.188) ％,(3.017 ± 0.356) ％,with statistically significant differences (F =77.660,P ＜ 0.001;F=32.200,P =0.001),further paired comparison,the apoptosis rate of Notch1-shRNA group was significantly higher than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).Western blotting showed that the expressions of PCNA,Bcl-2 and Hes-1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were statistically significant (F =155.343,P ＜ 0.001;F =22.576,P =0.002;F =70.108,P＜0.001),and the expressions of PCNA,Bcl-2 and Hes-1 in the three groups in SPC-A-1 cells were statistically significant (F =49.419,P ＜0.001;F =28.090,P =0.001;F =12.040,P =0.007).Further paired comparison,the expressions of PCNA,Bcl-2 and Hes-1 in Notch1-shRNA group were significantly lower than those in Nc-shRNA group in the two kinds of cells,and the differences were statistically significant (all P ＜0.05).Conclusion Targeted silencing of Notch1 can reduce the proliferation activity of lung cancer stem cells and promote apoptosis,which may be related to the down-regulation of its downstream gene Hes-1.

Nursing experience of 6 patients with severe pneumonia caused by H7N9 avian influenza / 中国实用护理杂志

Objective To summarize the nursing points of 6 patients with severe pneumonia caused by H7N9 avian influenza. Methods Six patients with pneumonia caused by H7N9 avian influenza in central Sichuan region were admitted from January to April 2017. Under the guidance of avian influenza expert group of Sichuan province, careful respiratory nursing for avian influenza patients were carried out, including mechanical ventilation, sputum suction, chest physiotherapy, the prevention of ventilator associated pneumonia and respiratory function exercise. Results Four patients of 6 patients were discharged from the hospital with the efforts of all medical staff, and 2 patients died unfortunately. Conclusions The severe pneumonia caused by avian influenza has a high death rate. Because respiratory nursing is one of most important treatments for avian influenza patients, this article summarized nursing points in this article as a reference for medical staff.

Nursing experience of 6 patients with severe pneumonia caused by H7N9 avian influenza / 中国实用护理杂志

Objective To summarize the nursing points of 6 patients with severe pneumonia caused by H7N9 avian influenza. Methods Six patients with pneumonia caused by H7N9 avian influenza in central Sichuan region were admitted from January to April 2017. Under the guidance of avian influenza expert group of Sichuan province, careful respiratory nursing for avian influenza patients were carried out, including mechanical ventilation, sputum suction, chest physiotherapy, the prevention of ventilator associated pneumonia and respiratory function exercise. Results Four patients of 6 patients were discharged from the hospital with the efforts of all medical staff, and 2 patients died unfortunately. Conclusions The severe pneumonia caused by avian influenza has a high death rate. Because respiratory nursing is one of most important treatments for avian influenza patients, this article summarized nursing points in this article as a reference for medical staff.

Alteration of Wnt/β-catenin signaling pathway in type2 diabetic rats’ aorta and regulation of SIRT1 / 中国药理学通报

Aim To investigate the alteration of Wnt/β-catenin signaling and sirtuins 1 in type 2 diabetic rats’ aorta and clarify its role in the development of di-abetes aortic disease. Methods The type 2 diabetes rat model was established by injection of streptozocin after five-week of high fat diet. The rats were randomly divided into control group, DM model group of 2 weeks, 4 weeks, 8 weeks and 12 weeks. Fasting blood glucose ( FBG ) , total cholesterol ( TC ) , triglyceride ( TG) , high density lipoprotein-cholesterol( HDL-C) , low density lipoprotein- cholesterol ( LDL-C ) and fast-ing insulin( FINS) levels were tested. HE staining was used to observe the pathological changes of aortal struc-tures. The alteration of Wnt2, β-catenin, TCF4, SIRT1 and sFRP2 in aortawas determined by Western blot and RT-PCR. Results Compared with control group, TC, TG, LDL-C levels of type 2 diabetic rats were significantly increased, HDL-C levels were signif-icantly reduced( P0. 05). But the expression of TCF4 and SIRT1 was enhanced continuously in DM compared with control group while sFRP2 decreased in the duration of DM development. Conclusions Wnt/β-catenin signa-ling pathway was activated in diabetic aortal injury by regulation of SIRT1 via sFRP2 . Further researches on its mechanism of actionin DM aorta injury may find a new therapeutic target for the disease.

Changes of heart function and serum cardiac troponin I in early type 2 diabetic rats / 中国病理生理杂志

AIM:To observe the changes of heart function and the expression of serum cardiac troponin I ( cT-nI) in early type 2 diabetic rats, and to explore the role of cTnI in the development of type 2 diabetes and early diabetic cardiomyopathy .METHODS:The type 2 diabetes rat model was established by an injection of streptozotocin after high fat diet (5 weeks).The rats were randomly divided into control group , model group of 2 weeks, and model group of 4 weeks. M-mode echocardiography was performed for echocardiographic measurements .Fasting blood glucose ( FBG) , total choles-terol ( TC) , triglyceride ( TG) , high density lipoprotein-cholesterol ( HDL-C) , low density lipoprotein-cholesterol ( LDL-C) , fasting insulin ( FINS) and cTnI levels were tested .HE staining was used to observe the pathological changes of myo-cardial structures .The alteration of cTnI in myocardium was determined by Western blot .RESULTS:Compared with nor-mal group , the levels of TC , TG and LDL-C in type 2 diabetic rats were significantly increased , HDL-C levels were signifi-cantly reduced .Cardiac histological analysis revealed that type 2 diabetes induced cardiomyocytes degeneration and necro-sis.The expression of cTnI increased significantly in diabetic groups compared to control group , and that in model group of 4 weeks increased far more than that in model group of 2 weeks (P<0.05).CONCLUSION:The increased level of cTnI and the change of the heart function may be associated with the development diabetic cardiomyopathy .These changes are valuable for the early clinical diagnosis of myocardial injury in diabetic cardiomyopathy .

Alteration of Wnt/β-catenin signaling pathway in early diabetic rat myocardium / 中国药理学通报

Aim To investigate the alteration of Wnt/β-catenin signaling pathway in early diabetic rat myo-cardium and clarify its role in development of diabetic cardiomyopathy. Methods The diabetes mellitus ( DM) model was prepared by intraperitoneal injection of streptozotocin ( STZ, 60 μg · g-1 ) . The alteration of Wnt/β-catenin signaling pathway was determined by Western blot and immunohistochemistry. HE staining was used to analyze the change of myocardial pathologi-cal structure. Results Cardiac histological analyses revealed that DM induced cardiomyocyte degeneration and necrosis. Myocardial Wnt2, β-catenin and c-Myc were enhanced in 2 wk DM compared with control group while DKK1 showed no significant alteration. Conclusion Wnt/β-catenin signaling pathway is acti-vated in early diabetic myocardial injury. Further re-searches on its role in DM myocardium may find a new therapeutic target for diabetic cardiomyopathy.

Effects of oldhamianoside Ⅱ on the invasive ability of prostatic carcinoma cells / 国际肿瘤学杂志

Objective To observe the effects of oldhamianoside Ⅱ on the invasive 'ability of prostatic carcinoma DU-145 cells,and to investigate the potential mechanism.Methods After treatment of DU-145 cells with oldhamianoside Ⅱ,MTT assay was used to test the effect of oldhamianoside Ⅱ on the cell proliferation.The invasive ability was assessed with a transwell cell culture chamber.Immunocytochemistry stain was used to investigate the expression of matrix metalloproteinase(MMP-2)and CXC-chemokine receptor 4 (CXCR4).Results The proliferation of DU-145 cells was inhibited after treatment with oldhamianoside Ⅱ.In vitro invasion assay indicated that the cells moved through the membrane were(180.3 ± 14.6)/field in the control group,while decreased to(100.4 ±1.5),(80.2±2.7)and(60.1 ±5.3)/field in 0.25,0.5 and 1μg/mL oldhamianoside Ⅱ treated DU-145 cells respectively.Comparing to control group,the cells of oldhamianoside Ⅱ groups moved through the membrane were decreased remarkably(P ＜0.05).Immunocytochemistry showed that the expression of MMP-2 and CXCR4 were decreased of oldhamianoside Ⅱ groups.Conclusions Oldhamianoside Ⅱ can inhibit the invasiveness of DU-145 cells,which is related to the down regulation the expression of MMP-2 in DU-t45 cells.Oldhamianoside Ⅱ can inhibit the prostate cancer cells metastasis to the bone,which is related to the down regulation the expression of CXCR4 in DU-145 cells.

Study on the Pharmacological Activity of Soybean Stem Soyasaponin / 中国中医药信息杂志

Objective To study the pharmacological activity of soybean stem soyasaponin. Methods Normobaric hyporia and exhaustive swimming in mice and osteoporosis rats model were used to observe the pharmacological effects of soybean stem soyasaponin. Results Soybean stem soyasaponin 150 mg/kg significantly prolonged the survival time in hyporia and the swimming time of mice and 100 mg/kg in rats improved the syomptom of osteoporosis models. Conclusion Soybean stem soyasaponin has obvious effect on anti-exhaustion and increasing tolerance of oxygen deficit in mice and significant therapy effect on osteoporosis rats.