This study aims to explore the effects of Shenqi Dihuang Decoction on high-glucose induced ferroptosis and the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4) axis in human renal tubular epithelial cells(HK-2) and to clarify the underlying mechanism. The cell injury model was established by exposing HK-2 to high glucose, and the Shenqi Dihuang Decoction-medicated serum was prepared. The optimal concentration and intervention time of Shenqi Dihuang Decoction were determined. HK-2 were divided into normal, high glucose, and low-, medium-, and high-dose Shenqi Dihuang Decoction groups. After interventions, the cell proliferation rate in each group was determined and the cell morphology and mitochondrial ultrastructure were observed. Then, the levels of intracellular reactive oxygen species(ROS), ferrous ion(Fe~(2+)), glutathione(GSH), and malondialdehyde(MDA) and the protein levels of Nrf2, HO-1, GPX4, and xCT were measured. The optimal concentration and intervention time of Shenqi Dihuang Decoction-medicated serum were determined to be 10% and 24 h, respectively. Compared with the high glucose group, high-dose Shenqi Dihuang Decoction promoted the proliferation of HK-2. The cells in the low-, medium-, and high-dose Shenqi Dihuang Decoction groups presented tight arrangement, an increased cell count, improved morphology from a spindle-fiber shape to a cobblestone shape, and improved morphology and structure of mitochondrial membrane and cristae, compared with those in the high glucose group. Meanwhile, all the doses of Shenqi Dihuang Decoction inhibited ROS elevation to mitigate the peroxidation damage, lowered the Fe~(2+) and MDA levels and elevated the GSH level to inhibit lipid peroxidation, and activated the antioxidant pathway to upregulate the protein levels of Nrf2, HO-1, xCT, and GPX4. In conclusion, Shenqi Dihuang Decoction-medicated serum can inhibit high-glucose induced ferroptosis of HK-2 in vitro, which involves the antioxidant effect and the activation of the Nrf2/HO-1/GPX4 pathway.

This study aims to explore the effects of Shenqi Dihuang Decoction on high-glucose induced ferroptosis and the nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4) axis in human renal tubular epithelial cells(HK-2) and to clarify the underlying mechanism. The cell injury model was established by exposing HK-2 to high glucose, and the Shenqi Dihuang Decoction-medicated serum was prepared. The optimal concentration and intervention time of Shenqi Dihuang Decoction were determined. HK-2 were divided into normal, high glucose, and low-, medium-, and high-dose Shenqi Dihuang Decoction groups. After interventions, the cell proliferation rate in each group was determined and the cell morphology and mitochondrial ultrastructure were observed. Then, the levels of intracellular reactive oxygen species(ROS), ferrous ion(Fe~(2+)), glutathione(GSH), and malondialdehyde(MDA) and the protein levels of Nrf2, HO-1, GPX4, and xCT were measured. The optimal concentration and intervention time of Shenqi Dihuang Decoction-medicated serum were determined to be 10% and 24 h, respectively. Compared with the high glucose group, high-dose Shenqi Dihuang Decoction promoted the proliferation of HK-2. The cells in the low-, medium-, and high-dose Shenqi Dihuang Decoction groups presented tight arrangement, an increased cell count, improved morphology from a spindle-fiber shape to a cobblestone shape, and improved morphology and structure of mitochondrial membrane and cristae, compared with those in the high glucose group. Meanwhile, all the doses of Shenqi Dihuang Decoction inhibited ROS elevation to mitigate the peroxidation damage, lowered the Fe~(2+) and MDA levels and elevated the GSH level to inhibit lipid peroxidation, and activated the antioxidant pathway to upregulate the protein levels of Nrf2, HO-1, xCT, and GPX4. In conclusion, Shenqi Dihuang Decoction-medicated serum can inhibit high-glucose induced ferroptosis of HK-2 in vitro, which involves the antioxidant effect and the activation of the Nrf2/HO-1/GPX4 pathway.

Introduction: The high glucose changes caused by diabetes mellitus (DM) can damage the vascular system. Astragaloside IV (AS-IV) can improve diabetes and promote angiogenesis. Exosomes (EXOs) help to carry specific drugs into cells efficiently. However, whether AS-IV loaded EXOs (AS-IV EXOs) can improve damaged endothelial cells through miR-214 remains to be determined. Material and methods: We prepared and identified AS-IV EXOs derived from endothelial progenitor cells (EPCs) and high glucose stimulated endothelial cell models to investigate whether AS-IV EXOs can improve damaged endothelial cells through miR-214. We used a transmission electron microscope (TEM) and DAPI staining to identify the morphology and characteristic expression of EPCs and EXOs, and then prepared AS-IV EXOs. Cell function tests were performed to detect the cloning, proliferation, and migration capabilities of cells. Western blot (WB) and real-time quantitative polymerase chain reaction (qRT-PCR) were used to assess the expression level of Tie-2, Ang-1, and PI3K/Akt-related protein. Results: The DAPI staining results showed that inducing human umbilical vein endothelial cells (HUVECs) could effectively absorb AS-IV EXOs. The results of plate clone formation assay, CCK-8, cell adhesion, and transwell assay of HUVECs stimulated by high glucose showed that AS-IV EXOs had a damage relief effect. By the detection of WB and qRT-PCR, it was found that AS-IV EXOs promoted the expression of miR-214 and proteins related to blood vessel growth. After transfection of miR-214 to pre-treat HUVECs under high glucose stimulation, AS-IV EXOs promoted the tube formation of HUVECs by regulating the level of miR-214. Conclusions: By promoting the expression of miR-214, AS-IV EXOs significantly improved the activity and tubularization of HUVECs under high glucose stimulation.

目的 探讨菝葜皂苷元(sarsasapogenin,SAR)对人结直肠癌细胞HT-29增殖、凋亡和自噬的影响.方法 将HT-29细胞分为对照组(DMEM培养液)和SAR组(DMEM培养液+DMSO+10、20、30、40、50、60 mg/L SAR溶液);分别采用MTT法、流式细胞术、TUNEL染色及MDC染色检测细胞的增殖、凋亡及自噬的变化情况;用Western blot法检测细胞中Caspase-3、Caspase-9、Be-clin-1及LC3B蛋白表达水平.结果 经SAR处理后,HT-29细胞的增殖能力明显降低(P<0.05),凋亡和自噬水平明显增加(P<0.05);与对照组相比,SAR组细胞Caspase-3、Caspase-9、Beclin-1及LC3B蛋白表达水平显著上调(P<0.05).结论 SAR能够抑制结直肠癌细胞增殖,诱导细胞凋亡和细胞自噬的发生,其机制可能与上调Caspase-3、Caspase-9、Beclin-1及LC3B蛋白表达水平有关.

目的 研究红车轴草总黄酮对高糖诱导的大鼠胰岛β细胞损伤的影响,及其潜在的分子机制.方法 本实验分为对照组,模型组,低、中、高剂量实验组,miR-NC组,miR-99a-3p组,anti-miR-NC,anti-miR-99a-3p组,模型+miR-NC组,模型+miR-99a-3p组,模型+si-NC组,模型+si-CD36组,高剂量实验+anti-miR-NC组和高剂量实验+anti-miR-99a-3p组.对照组给予5.5 mmol·L-1葡萄糖处置;模型组给予25 mmol·L-1葡萄糖处置;低、中、高剂量实验组分别给予12.5,25.0和50.0 mg·L-1红车轴草总黄酮和25 mmol·L-1葡萄糖处置;miR-NC组、miR-99a-3p组、anti-miR-NC组、anti-miR-99a-3p组分别转染miR-NC、miR-99a-3p、anti-miR-NC、anti-miR-99a-3p;模型+miR-NC组、模型+miR-99a-3p组、模型+si-NC组、模型+si-CD36组均给予25 mmol·L-1葡萄糖处理,并分别转染miR-NC、miR-99a-3p、si-NC、si-CD36;高剂量实验+anti-miR-NC组、高剂量实验+anti-miR-99a-3p组均给予50.0 mg·L-1红车轴草总黄酮+25 mmol·L-1葡萄糖,并分别转染anti-miR-NC或anti-miR-99a-3p.用流式细胞术测定细胞凋亡率,用定量实时聚合酶链反应检测细胞中miR-99a-3p和CD36 mRNA的表达水平.结果 模型组INS-1细胞中CD36 mRNA(2.74±0.27 vs 1.01±0.08)和细胞凋亡率[(27.63±2.71)％vs(5.69±0.58)％]均较对照组显著升高,miR-99a-3p(0.38±0.03 vs 1.00±0.09)较对照组显著降低,差异均有统计学意义(均P<0.05);与模型组比较,低、中、高剂量实验组的上述结果相反.模型+miR-99a-3p组的细胞凋亡率[(12.48±1.19)％vs(26.84±2.41)％]较模型+miR-NC组显著降低,miR-99a-3p表达量(0.79±0.07 vs 0.34±0.03)较模型+miR-NC组显著升高,差异均有统计学意义(均P<0.05).模型+si-CD36组INS-1细胞中CD36 mRNA(1.35±0.12 vs 2.81±0.26)及细胞凋亡率[(13.54±1.32)％vs(25.87±2.52)％]均较模型+si-NC组显著降低,差异均有统计学意义(均P<0.05).高剂量实验+an-ti-miR-99a-3p组INS-1细胞中miR-99a-3p(0.43±0.14 vs 0.88±0.06)较高剂量实验+anti-miR-NC组显著降低,CD36 mRNA(2.51±0.22 vs 1.41±0.13)及细胞凋亡率[(20.45±2.03)％vs(10.56±1.02)％]均较高剂量实验+anti-miR-NC组显著升高,差异均有统计学意义(均P<0.05).结论 红车轴草总黄酮可能通过调控miR-99a-3p/CD36以减轻高糖对大鼠胰岛β细胞INS-1的损伤,进而抑制细胞的凋亡.

目的 本研究旨在运用网络药理学方法,预测开心散活性成分的抗抑郁药理作用机制.方法 通过TCMSP、ETCM获取开心散的成分和靶点,运用Genecard、OMIM筛选抑郁症相关的蛋白,通过STRING构建蛋白相互作用网络,使用Cytoscape进行靶点的拓扑分析、MCODE聚类分析,使用R语言进行GO和KEGG通路富集分析,最后使用MOE软件进行分子对接验证.结果 网络分析表明,开心散中25个活性成分与其发挥抗抑郁作用密切相关,该机制涉及到51个靶点(包括ACHE、HTR3A、DRD2等)以及24条通路;分子对接结果显示化合物aposcopolamine、perlolyrine、hederagenin与PPARG、ACHE、SLC6A4、DRD2靶点均具有较好的结合能力.结论 开心散可能主要通过神经活性受体配体相互作用、羟色胺突触、cAMP通路等相关靶标起到抗抑郁的功效,该研究为一步揭示开心散治疗抑郁症的潜在活性成分及其可能的作用机制提供了思路.

目的 基于网络药理学及分子对接初步探讨四妙丸治疗痛风的作用机制.方法 通过中药系统药理数据库和分析平台(TCMSP)和中药分子机制的生物信息学在线分析工具(BATMAN-TCM)数据库筛选四妙丸所含的活性化合物及作用靶点;检索GeneCards数据库获取痛风相关的疾病靶点;将药物和疾病靶点导入Bioinformatics&Evolutionary Genomics取交集,即药物(四妙丸)与疾病(痛风)的共同靶点,并利用Cytoscape 3.7.1软件构建四妙丸-化合物-靶点-痛风网络图;通过STRING数据库和R软件分别绘制PPI网络图和进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析;另外选取部分化合物和靶点通过MOE软件进行分子对接验证;最后通过单钠尿酸盐(MSU)结晶诱导人髓系白血病单核细胞(THP-1)复制痛风细胞模型以进一步验证上述结果 .结果 本研究共筛选出四妙丸相关活性化合物65个,药物相关靶点724个,四妙丸与痛风的共同靶点20个;网络图提示槲皮素和山柰酚,环加氧酶1(PTGS1)可能分别是四妙丸治疗痛风的有效化合物和靶点;GO富集分析提示四妙丸治疗痛风主要与转录辅因子、转录因子、增强子、肌动蛋白、核转录因子(NF-κB)等有关;KEGG富集分析显示四妙丸可能通过AGE-RAGE、NF-κB、MAPK8和TNF等多条信号通路治疗痛风;分子对接结果显示槲皮素、山柰酚、汉黄芩素、小檗碱与MAPK8有良好对接活性;细胞实验结果表明,槲皮素能够抑制MSU诱导THP-1细胞后MAPK8蛋白的表达.结论 四妙丸治疗痛风呈现多分子、多靶点、多通路的协同作用机制,为后续深入探讨四妙丸治疗痛风的分子机制提供理论依据.

[目的]通过随机对照临床试验评估白虎汤合增液汤治疗热盛阴虚型2型糖尿病患者的临床疗效.[方法]纳入我院2016年4月至2018年4月之间确诊的112例热盛阴虚型2型糖尿病患者,按照1:1随机分为试验组和常规用药对照组,对照组采用吡格列酮片治疗,试验组在此基础上联合使用白虎汤合增液汤治疗,两组均持续治疗8周,8周后评估两组患者临床疗效、 空腹血糖(fasting blood glucose,FBG)、 餐后2h血糖(2h postprandial blood glucose,2hPBG)、糖化血红蛋白(glycated hemoglobin,HbAlc)、血清总胆固醇(total cholesterol,TC)、血清甘油三脂(triglyceride,TG)和中医临床证候积分的变化.[结果]治疗8周后,试验组总体疗效(96.4％)明显优于对照组(78.6％),差异具有统计学意义(P<0.05);试验组的中医临床证候积分明显好于对照组,差异具有统计学意义(P<0.05);试验组的FBG、2hPBG、HbAlc、TC、TG显著改善,并明显低于对照组,差异具有统计学意义(P<0.05);试验组的不良反应率(3.57％)明显低于对照组(14.3％),差异具有统计学意义(P<0.05).[结论]采用白虎汤合增液汤联合吡格列酮片治疗热盛阴虚型2型糖尿病患者可以更有效地改善糖尿病患者的临床症状,降低血糖及血脂,提高患者生活质量.

目的 研究自体脂肪干细胞局部注射对糖尿病足创面愈合及细胞因子、Wnt/β-catenin通路的影响.方法 前瞻性选择2016年2月至2017年12月期间湖南中医药大学第一附属医院收治的糖尿病足患者,随机分为接受自体脂肪干细胞局部注射联合湿润烧伤膏换药的观察组,接受湿润烧伤膏换药的对照组.观察治疗后2周、3周、4周时的创面愈合率以及治疗过程中的创面感染率、创面愈合时间,测定2周后创面中细胞因子的含量、Wnt/β-catenin通路分子的表达量.结果 观察组患者治疗2周、3周、4周后的创面愈合率高于对照组,治疗过程中的感染率低于对照组、平均愈合时间短于对照组(P<0.05);观察组患者治疗后创面中血管内皮生长因子(VEGF)、表皮生长因子(EGF)、转化生长因子-β1(TGF-β1)的含量以及Wnt1、β连环蛋白(β-catenin)的mRNA表达量均高于对照组,白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)、血管细胞黏附分子-1(VCAM-1)、肿瘤坏死因子-α(TNF-α)的含量以及糖原合成激酶-3β(GSK-3β)的mRNA表达量均低于对照组(P<0.05).结论 自体脂肪干细胞局部注射能够促进糖尿病足创面的修复并调节细胞因子分泌、Wnt/β-catenin通路激活.

Objective To investigate the expression of galectin-3 in wounds and sera of patients with diabetic foot ulcer(DFU) so as to clarify the correlation between galectin-3 and DFU. Methods Fifty DFU patients with Wagner 1-2 stages and 20 wounded patients with normal blood glucose level(serving as the control group)were enrolled to obtain their tissues from the edge of the foot wound and sera from January 2015 to March 2016. The serum concentration of galectin-3 was detected by ELISA,and the expression of galection-3 in the tissues was detected by Western-blot. Results ELISA results showed that the serum con-centration of galectin-3 was higher in the DFU group than in the wounded control group((5.66±2.76)ng/mL vs.(4.25±1.94) ng/mL,P=0.04). Western-blot results indicated that the expression of galectin-3 in tissues from wound edge of the DFU group was enhanced. Conclusions Galectin-3 is highly expressed in the patients with DFU,and its high expression may contribute to the development of DFU.

Objective To investigate the expression of long non-coding RNA NONHSA1254644 in lung adenocarcinoma and its clinical significance.Methods 99 cases of lung adenocarcinoma and the adjacent tissues as well as clinical data was collected.The expression level was detected by quantitative realtime polymerase chain reaction (qRT-PCR).Then the relationship between the expression level and the clinical parameters was analyzed.Cell counting kit-8 (CCK8) was used to investigate its effect of lung adenocarcinoma cell line A549 on proliferation.Results The expression of NONHSA1254644 was down-regulated in lung adenocarcinoma,and its expression was positively correlated with the differentiation of patients.The overexpression of NONHSA1254644 could inhibit the proliferation of A549 cells.Conclusions NONHSA1254644 is involved in the development and progression of lung adenocarcinoma.

Aim The aim of this study was to establish the accuracy of transthoracic echocardiography (TTE) in diagnosis of acute aortic dissection type Stanford A.Methods A total of 35 cases of acute aortic dissection type Stanford A were retrospectively analyzed,who underwent both transthoracic echocardiography and CTA.Results The diagnostic accuracy of TTE and CTA of acute aortic dissection type Stanford A were 91.3％ and 100％,respectively.However,TTE could show aortic valve abnormalities,heart function,pericardial effusion.Conclusion TTE is a reliable method for diagnosis of acute aortic dissection type Stanford A.The pivotal role of TTE in the evaluation of the patients with suspected acute type A aortic dissection in emergency room setting.

Amyloid β-protein precursor (APP) is a major molecule inducing neurodegeneration in diabetic patients.To explore the effects of high-glucose cultivation on the injury of primary hippocampal neurons and the expression of APP,the primary hippocampal neurons were separated from the embryonic day 16 (E16) SD rats and then cultured in the presence of 25 mmol/L,50 mmol/L,75 mmol/L and 100 mmol/L glucose,respectively.The cell morphology was observed with an inverted microscope and the expression of APP protein was detected by Western-blot.The results indicated that,following exposure to high concentrations of glucose,the axons of hippocampal neurons became swelling and the synapses gradually grew shorter,while the expression of APP increased.The results suggested that APP might play important roles in high glucoseinduced hippocampal neuron damage and that glycemic control would be potentially beneficial for the protection of neuronal cells.

Objective To evaluate the value of two￣dimension speckle tracking imaging in assessing left ventricular global systolic function in diastolic heart failure in patients with essential hypertension. Methods The study included 65 patients with essential hypertension and preserved left ventricular systolic function (LVEF >50%),which were divided into patients with diastolic heart failure (n=26,DHF group) and those without heart failure(n=39,NHF group).26 age matched healthy controls were selected randomly.Two￣dimensional echocardiographic images of the left ventricle were acquired in apical four￣,two￣chamber,long￣axis views,and parasternal short￣axis at the basal,mid,and apical levels.Global longitudinal strain (GLS), global radial strain(GRS), global circumferential strain(GCS) were quantified using a novel speckle tracking sys￣tem.Global parameters were investigated among the groups. Results Compared with the control group,the value of GLS, GRS,GCS were significantly reduced in DHF group and NHF group( P<0 05);Compared with the NHF group,the value of GLS,GRS,GCS were significantly reduced in DHF group ( P<0 05) . Conclusions 2D￣STI can evaluate 1eft ventricular global systolic function in diastolic heart failure in patients with essential hypertension,and has potential applications in clini￣cal practice.

目的:探讨对比BNP水平、E/A和E/E'评价高血压伴舒张性心力衰竭的严重性,为确定E/E'在临床上能否作为鉴定高血压伴舒张性心力衰竭的非创伤性指标奠定基础.方法:80例原发性高血压患者分为舒张性心力衰竭组(n=35)和非舒张性心力衰竭组(n=45),测量舒张早期二尖瓣血流速度(E)、舒张晚期二尖瓣血流速度(A)和舒张早期二尖瓣环运动速度(E'),同时测量BNP水平.随机选择30名健康人为正常对照组.结果:BNP与E/A比值间无相关性,BNP与E/E'比值正相关(P＜0.01),E'在DHF组明显低于NHF组.BNP、E/E'在所有患者中随着心力衰竭严重程度的增加而增加.结论:E/E'比值可作为评估有症状舒张性心力衰竭等级(伴有高血压)的灵敏候选指标之一.