Glucocorticoid reduces the efficacy of afatinib on the head and neck squamous cell carcinoma

Glucocorticoids(GC)are widely used to counter the adverse events during cancer therapy;nonetheless,previous studies pointed out that GC may reduce the efficacy of chemotherapy on cancer cells,especially in epidermal growth factor receptor(EGFR)-targeted therapy of head and neck squamous cell carcinoma(HNSCC)remaining to be elucidated.The primary aim of the present study was to probe into the GC-induced resistance of EGFR-targeted drug afatinib and the underlying mechanism.HNSCC cell lines(HSC-3,SCC-25,SCC-9,and H-400)and the human oral keratinocyte(HOK)cell lines were assessed for GC receptor(GR)expression.The promoting tumor growth effect of GC was evaluated by the CCK-8 assay and flow cytometry.Levels of signaling pathways participants GR,mTOR,and EGFR were determined by quantitative polymerase chain reaction and western blotting.GC increased the proliferation of HNSCC cells in a GR-dependent manner and promoted AKT/mTOR signaling.But GC failed in counteracting the inhibition of rapamycin in the mTOR signaling pathway.Besides,GC also induced resistance to EGFR-targeted drug afatinib through AKT/mTOR instead of the EGFR/ERK signaling pathway.Thus,GCs reduce the efficacy of afatinib on HNSCC,implicating a cautious use of glucocorticoids in clinical practice.

Subjective Global Assessment as a Pre-Operative Nutrition Status Screening Tool for Head and Neck Cancer Patients of a Tertiary Health Care Setting

Introduction: In India, 57% of patients with head and neck cancers are documented with nutritional compromise. Active nutritional support has been shown to improve outcomes and reduce the cost of treatment in severely malnourished patients. The assessment of nutritional status should be a priority when initiating medical nutrition therapy. We evaluated the agreement between Subjective and Objective evaluation of pre-operative nutrition status of head and neck cancer patients in a tertiary cancer centre. Methods: Two hundred and thirty seven head and neck cancer patients who underwent surgery were eligible. The patients included both males (147) and females (90) with age varying between 23 - 88 years. All patients were screened for pre-operative nutrition status objectively as well as subjectively. The association of pre-operative SGA scores (A, B and C) subjectively, and PNS score (0, 1, 2) objectively were tested for statistical significance. Results: The cancer sites included tongue in 82, buccal mucosa in 30, thyroid in 28, alveolus in 18, glottis in 10, RMT in 10, nasal cavity in 9, FOM in 8. The pre-operative nutrition status based on subjective scores are A in 156 (65.8%), B in 75 (32%) and C in 6 (2.5%). The objective parameters obtained on the basis of BMI, % weight loss, PNI and S. albumin values are PNS 0 in 161 (67.9%), PNS 1 in 71 (30%) and PNS 2 in 5 (2.1%) patients. As the kappa coefficient p-0.56, there is moderate agreement between the pre-operative nutrition status subjectively as well as objectively. Conclusion: Subjective global assessment is a simple and inexpensive way to screen the pre-operative nutrition status when compared to the other objective assessment tool. SGA has moderate agreement with expensive and complicated objective assessment tools. So it can be a reliable tool for assessing the pre-operative nutrition status.

Effect of Gubi Tongxiao granules on osteoblast injury induced by glucocorticoid

Objective:To explore the mechanism of Gubi Tongxiao granule in treating osteoblast injury induced by glucocorticoid.Methods:Mouse osteogenic precursor cell MC3T3-E1 was cultured in vitro,the control group was conventionally cultured,the model group was treated with dexamethasone(10^(-5) M,10^(-6) M,10^(-7) M)solution with gradient concentration to induce injury,and the experimental group was added with Gubi Tongxiao granule drug-containing serum to intervene culture on the basis of the model group.Cell viability was detected by CCK-8 method,autophagy and apoptosis were detected by MDC/PI staining,Beclin-1,Bcl-2 and Bax expression levels were detected by Western-blot,and the expression of target genes in each group was analyzed by RT-qPCR.Results:Compared with the control group,the activity of osteoblasts in the model decreased significantly.The effect of dexamethasone on autophagy and apoptosis of MC3T3-E1 cells was time-dependent and dose-dependent(P<0.05).The autophagy marker Beclin-1 increased at low doses of dexamethasone(10^(-7) or 10^(-6) M)and decreased at high doses(10^(-5) M).Compared with model group,Beclin-1 and Bax mRNA expression in experimental group decreased(P<0.01),and Bcl-2 mRNA expression was significantly up-regulated(P<0.01).In addition,the results of semi-quantitative analysis of apoptosis staining showed that autophagy and apoptosis were inhibited in different degrees(P<0.01).Conclusion:Gubi Tongxiao granule has a protective effect on osteoblast injury caused by dexamethasone,and can effectively reduce autophagy and apoptosis induced by glucocorticoid,which may be one of the mechanisms of maintaining bone mass and delaying the occurrence and development of femoral head necrosis.

Bone palsy eliminates granules to regulate Wnt/PI3K-AKT signaling pathway and intervene in hormonal osteonecrosis of the femoral head in rabbits

Objective:To investigate the therapeutic effect of osteopathic osteopathy on hormonal osteonecrosis in rabbits by regulating the Wnt/PI3K-AKT signaling pathway.Methods:Thirty clean-grade New Zealand rabbits were randomly divided into control group,model group and experimental group,and all except for the control group,allogeneic serum combined with glucocorticoids were used to establish animal models of simulated hormonal osteonecrosis of the femoral head.After successful modeling,the experimental group was given the model group and the control group the same amount of normal saline,and the experimental group was given bone palsy granule concentrate according to the equivalent dose of humans and rabbits,once a day for 8 weeks,and after intraperitoneal anesthesia,blood was collected from the extracorporeal heart and the serum ALP and TGF-β1 levels were detected by immunoenzyme-linked adsorption.The experimental animals were sacrificed,,and the bilateral femoral heads of rabbits were removed for hematoxylin-eosin staining,and the apoptosis of cells in the femoral head area was observed by TUNEL staining.RT-qPCR analyzed the expression of Bax and Bcl-2 genes,and Western blot detected the expression levels of pathway proteins Wnt,β-catenin,GSK-3β,AKT and p-AKT.Results:Compared with the control group,the expression levels of ALP and TGF-β1 in rabbit bone tissue in the model group decreased significantly(P<0.05),the bone trabeculae were sparse and thinned,the number of hollow bone fosses and the number of apoptosis-positive cells increased significantly(P<0.05),the expression of pro-apoptotic gene Bax was increased,and the expression of anti-apoptotic gene Bcl-2 was inhibited(P<0.05).After the intervention and treatment of bone palsy granules,the hollow bone depression and apoptosis of rabbit bone tissue in the experimental group were effectively improved(P<0.05).In addition,compared with the model group,the osteopathic elimination particles could upregulate the expression of ALP and TGF-β1(P<0.05),promote the expression of pathway proteins Wnt andβ-catenin,increase the phosphorylation level of AKT,and downregulate the expression of GSK-3β(P<0.05).Conclusion:Under glucocorticoids,osteoblast development dysfunction and apoptosis increase.Bone palsy granules can regulate the two signaling pathways of Wnt/PI3K-AKT,promote osteoblast development and inhibit apoptosis,effectively maintain bone metabolic balance,prolong the collapse time of the femoral head,and further treat osteonecrosis of the femoral head.

三七有效成分调控激素性股骨头坏死的相关信号通路

背景:激素性股骨头坏死是骨科领域难治和难愈性疾病,尚无确切研究观点完整解释其病理机制。随着三七有效成分干预多种疾病相关信号通路的研究增加,三七有效成分通过调控激素性股骨头坏死相关信号通路治疗该病逐渐成为时下研究热点。目的:系统总结分析近年来激素性股骨头坏死病理机制研究文献和三七有效成分调控该病相关信号通路的相关文献内容,为后续研究三七有效成分治疗该病提供参考。方法:检索中国知网及万方数据库,中文检索词为“糖皮质激素,激素性股骨头坏死,病理机制,信号通路,三七,有效成分”等;检索PubMed数据库,英文检索词为”Glucocorticoid,steroid associated necrosis of the femoral head,Pathological mechanism,signaling pathway,Panax notoginseng,active ingredient”等,筛选激素性股骨头坏死病理机制相关文献和三七有效成分调控激素性股骨头坏死相关信号通路文献,最终共纳入63篇文献。结果与结论:①三七主要成分包括三七总皂苷、人参皂苷、三七皂苷、槲皮素和山柰酚等。②在调控相关通路方面,三七总皂苷、人参皂苷Rb1和槲皮素作用于转化生长因子β/骨形态发生蛋白通路,能促进骨修复和血管新生;三七总皂苷、人参皂苷CK和山柰酚作用于Wnt/β-catenin通路,可促进成骨分化和脂质代谢;三七总皂苷及三七皂苷R1/R2作用于MAPK通路,抑制破骨和促进骨修复;三七总皂苷、人参皂苷Rb2及槲皮素作用于RANKL/RANK/骨保护蛋白通路,可抑制破骨增殖并促进成骨分化;三七总皂苷、槲皮素及山柰酚作用于缺氧诱导因子1α通路,能修复血管损伤和促进成骨。③三七皂苷R1、槲皮素联合羟基磷灰石纳米颗粒、三七总皂苷联合聚乙烯-L-乳酸等生物材料治疗激素性股骨头坏死具有良好的研究前景。④三七有效成分可通过多种机制调控激素性股骨头坏死相关信号通路,对该病起到积极的干预作用,但目前相关研究深度和广度不足,未来应基于现有的机制研究,深入探究三七调控该病不同通路的具体机制及通路间相互作用,将有利于运用三七有效成分治疗激素性股骨头坏死的多元发展。

全球心血管疾病学术影响力评价(CAPE)体系的方法探索

目的:构建全球心血管疾病学术影响力评价(Cardiovascular Academic Performance Evaluation,CAPE)体系,并对全球医疗机构心血管疾病学术影响力进行排名。方法:由心血管疾病领域人员和图书情报人员提取医学主题词表(MeSH)、Embase同义词表、国际疾病分类(ICD)中心血管疾病相关术语,建立心血管术语词表(即阜外词表),设定缺血性心脏病、高血压、血管疾病、心律失常、肺血管疾病、心力衰竭、先天性心脏病、心肌病和瓣膜性心脏病等9个亚学科,实现亚学科分类、心血管术语词、入口词的映射。检索Web of Science、PubMed、Scopus数据库2016年1月1日至2022年12月31日的心血管领域论文,并对元数据进行去重融合;应用阜外词表对论文中主题词、题目、关键词、摘要等4个位置的内容进行检索匹配,生成论文“位置-心血管术语词-频次”的信息表,计算每一篇论文心血管学科相关度积分及亚学科积分。对论文的机构名、学者名进行规范化清洗,基于心血管强相关(相关度积分≥4分)原创性论文开展全球心血管疾病领域学术影响力的评价。针对全国医疗机构(心血管)的科技评价,还采集了国家自然科学基金项目、授权发明专利、获奖成果、科研平台、注册试验[国家药品监督管理局药品审评中心(CDE)药物临床试验登记数据]等数据。结果:检索到全球2016~2022年心血管领域论文1545103篇,剔除摘要、书籍、传记、新闻、视频、音频、撤稿、更正声明后为1178019篇,使用阜外词表标引“心血管强相关”论文518058篇。还采集到11143项心血管相关国家自然科学基金项目、19382项心血管相关有效授权发明专利、103项心血管相关国家级获奖成果、24个心血管相关国家科研平台、2084项CDE心血管相关药物临床试验登记等数据。经组织心血管疾病及其亚学科专家团队针对各自亚学科论文进行核对和验证,改进学科分类规则,最终锁定11项指标构建了心血管疾病和9个亚学科领域的综合评价指标体系。2016~2022年,中国发表心血管疾病研究论文数量增幅为123.5%,共发表约7.68万篇,平均每天约30篇,仅次于美国(约11.41万篇),居世界第二位。但中国期刊引证报告(JCR)分区和中国科学院分区论文占比仅居世界第八位。在2020~2022年全球医疗机构心血管原创研究论文影响力综合评估中,仅2家中国医疗机构进入前20名。结论:CAPE项目基于2016~2022年多源数据基础,首次探索建立了心血管领域机构学术影响力综合评价指标体系。

激素性股骨头坏死滑膜病变分子机制的实验研究

目的:探讨激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)滑膜病变的分子机制。方法:(1)转录组测序及生物信息学分析。收集9例接受全髋关节置换术的患者(SONFH患者3例、髋骨关节炎患者3例和股骨颈骨折患者3例)术中切除的髋关节滑膜组织,进行转录组测序和生物信息学分析,筛选SONFH滑膜病变核心基因。(2)滑膜组织检测。根据标本来源将髋关节滑膜组织分为SONFH组、髋骨关节炎组和股骨颈骨折组。观察各组髋关节滑膜组织的组织形态,采用实时定量PCR检测3组髋关节滑膜组织中SONFH滑膜病变核心基因的mRNA相对表达量,采用蛋白质印迹法和免疫组织化学染色法检测3组髋关节滑膜组织中SONFH滑膜病变核心基因的蛋白相对表达量。采用免疫荧光染色法检测SONFH滑膜病变的靶细胞。(3)细胞验证。培养大鼠滑膜成纤维细胞,构建滑膜炎细胞模型。采用实时定量PCR及蛋白质印迹法检测空白细胞(空白细胞组)和滑膜炎模型细胞(模型细胞组)中SONFH滑膜病变核心基因的表达。结果:(1)转录组测序和生物信息学分析结果。经对不同患者来源的髋关节滑膜组织进行差异基因分析,共筛选出1001个与SONFH髋关节滑膜病变相关的基因,这些基因与免疫反应和外泌体有关,其中干扰素调节因子(interferon regulatory factor,IRF)4和IRF7是SONFH髋关节滑膜病变的核心基因,两者均为参与Ⅰ型干扰素应答的关键转录因子。(2)滑膜组织形态观察结果。苏木素-伊红染色显示,股骨颈骨折组髋关节滑膜组织形态正常,无细胞增生、肥大或间质水肿;髋骨关节炎组髋关节滑膜组织细胞增生,有少量新生血管和细胞聚集;SONFH组髋关节滑膜组织细胞大量增殖和聚集,有新生血管。(3)滑膜组织中SONFH滑膜病变核心基因表达检测结果。SONFH组髋关节滑膜组织中IRF4、IRF7、干扰素-α(interferon-α,IFN-α)的mRNA相对表达量均高于股骨颈骨折组和髋骨关节炎组(P=0.000,P=0.000,P=0.000;P=0.001,P=0.000,P=0.036),髋骨关节炎组髋关节滑膜组织中IRF4、IRF7、IFN-α的mRNA相对表达量均高于股骨颈骨折组(P=0.000,P=0.000,P=0.000)。IRF4、IRF7、IFN-α蛋白在SONFH组髋关节滑膜组织中高表达,SONFH组髋关节滑膜组织中IRF4、IRF7、IFN-α的蛋白相对表达量均高于股骨颈骨折组和髋骨关节炎组(P=0.001,P=0.000,P=0.000;P=0.000,P=0.014,P=0.000),髋骨关节炎组髋关节滑膜组织中IRF4、IRF7、IFN-α的蛋白相对表达量均高于股骨颈骨折组(P=0.002,P=0.005,P=0.000)。(4)SONFH滑膜病变靶细胞检测结果。SONFH组髋关节滑膜组织中IRF7、钙黏附蛋白11(cadherin-11,CDH-11)的蛋白相对表达量均高于股骨颈骨折组和髋骨关节炎组(P=0.001,P=0.000;P=0.000,P=0.001),髋骨关节炎组髋关节滑膜组织中IRF7、CDH-11的蛋白相对表达量高于股骨颈骨折组(P=0.001,P=0.000)。滑膜成纤维细胞为SONFH滑膜病变的靶细胞。(5)细胞验证结果。模型细胞组IRF4、IRF7、IFN-α的mRNA和蛋白相对表达量均高于空白细胞组(P=0.001,P=0.002,P=0.000;P=0.001,P=0.007,P=0.000)。结论:SONFH滑膜病变与免疫炎症反应关系密切,滑膜成纤维细胞可能是SONFH滑膜病变的靶细胞,IRF4和IRF7可能是其潜在的靶点。

激素性股骨头坏死囊性变组织骨修复机制分析与活血通络胶囊含药血清对肥大软骨细胞成骨分化影响的实验研究

目的:分析激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)囊性变组织的骨修复机制,并观察活血通络胶囊含药血清对肥大软骨细胞成骨分化的影响。方法:(1)SONFH囊性变组织的骨修复机制分析。纳入6例采用全髋关节置换术治疗的SONFH患者,术后收集坏死股骨头6个,其中3个股骨头用于囊性变组织的单细胞转录组测序,采用生物信息学方法分析囊性变组织中的细胞分类、软骨细胞的细胞亚群、肥大软骨细胞参与的生物过程;另外3个股骨头用于囊性变组织的病理学检查,分别采用苏木素-伊红染色、阿利新蓝染色、番红O-固绿染色观察囊性变组织中软骨细胞、肥大软骨细胞的分布与特征,采用免疫组织化学染色观察囊性变组织中肥大软骨细胞标志蛋白Ⅹ型胶原α1(collagen typeⅩα1,Col10α1)、基质金属蛋白酶(matrix metalloproteinase,MMP)13和成骨分化相关蛋白Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)、骨桥蛋白(osteopontin,OPN)的表达情况。(2)活血通络胶囊含药血清对肥大软骨细胞成骨分化影响的分析。采用活血通络胶囊(活血通络胶囊粉末溶于蒸馏水)给大鼠灌胃,制备活血通络胶囊含药血清。取小鼠膝关节软骨,消化分离后进行培养。将小鼠软骨细胞接种于含有不同浓度的活血通络胶囊含药血清的完全培养基中,筛选活血通络胶囊含药血清的最佳干预浓度。取处于对数生长期的第1代小鼠软骨细胞,行肥大软骨细胞诱导成功后,分为成骨诱导组、5%含药血清干预组和对照组,成骨诱导组更换成骨诱导培养基培养,5%含药血清干预组更换含5%活血通络胶囊含药血清的成骨诱导培养基培养,对照组继续采用肥大诱导培养基培养;培养7 d后,分别采用碱性磷酸酶(alkaline phosphatase,ALP)染色和茜素红染色,观察细胞形态特征,分别采用实时定量PCR和蛋白质印迹法检测Ⅰ型胶原蛋白α1(collagenⅠα1,Col1α1)、ALP、RUNX2、OPN的mRNA相对表达量和ALP、RUNX2、OPN的蛋白相对表达量。结果:(1)SONFH囊性变组织单细胞转录组测序分析结果。共获得30224个细胞的单细胞转录组测序结果;细胞分类结果显示,囊性变组织中有8种细胞,包括T细胞、内皮细胞、成纤维细胞、软骨细胞、巨噬细胞、单核细胞、破骨细胞和肥大细胞;软骨细胞亚群分析共鉴定出5类亚群细胞,包括稳态软骨细胞、成纤维软骨细胞、炎症软骨细胞、肥大软骨细胞、前体肥大软骨细胞;差异基因富集分析结果显示,肥大软骨细胞主要参与细胞迁移正向调节、细胞分化、细胞外基质形成、骨化、细胞迁移和成骨细胞分化等生物过程。(2)SONFH囊性变组织的病理学检查结果。囊性变组织的边缘可见软骨细胞、肥大软骨细胞,且部分软骨组织呈现向骨小梁过渡的形态;Col10α1、MMP13、RUNX2、OPN在囊性变组织的边缘高表达。(3)活血通络胶囊含药血清干预肥大软骨细胞成骨分化的分析结果。经筛选,5%为活血通络胶囊含药血清的最佳干预浓度。ALP染色和茜素红染色结果显示,对照组无明显钙化结节,成骨诱导组有明显钙化结节,5%含药血清干预组钙化结节较成骨诱导组进一步增多;成骨诱导组细胞ALP、RUNX2、OPN、Col1α1的mRNA相对表达量和ALP、RUNX2、OPN的蛋白相对表达量均高于对照组(P=0.000,P=0.000,P=0.001,P=0.000;P=0.000,P=0.000,P=0.001),5%含药血清干预组细胞ALP、RUNX2、OPN、Col1α1的mRNA相对表达量和ALP、RUNX2、OPN的蛋白相对表达量均高于成骨诱导组(P=0.000,P=0.000,P=0.002,P=0.001;P=0.000,P=0.000,P=0.001)。结论:SONFH囊性变组织中存在肥大软骨细胞向成骨细胞分化的修复机制,而活血通络胶囊含药血清能够促进体外培养的肥大软骨细胞向成骨细胞分化。

中药调控成骨细胞铁死亡治疗激素性股骨头坏死

背景:有研究发现成骨细胞铁死亡可作为重要的发病机制诱导激素性股骨头坏死的发生与发展。随着祖国医学的发展,有学者发现某些中药单体、中药复方及中成药等可通过多种通路机制调控成骨细胞铁死亡,最终起到治疗激素性股骨头坏死的作用。目的:探讨成骨细胞铁死亡与激素性股骨头坏死的关系及中草药调控成骨细胞铁死亡治疗激素性股骨头坏死的作用机制,为激素性股骨头坏死的诊治提供新的思路。方法:以“铁死亡,激素性股骨头坏死,成骨细胞,中草药,糖皮质激素,铁代谢,活性氧,谷胱甘肽过氧化物酶”为中文检索词,以“ferroptosis,Hormonal necrosis of the femoral head,osteoblast,Chinese herbal medicine,glucocorticoid,iron metabolism,ROS,GPX4”为英文检索词,检索中国知网、Pub Med、万方及维普数据库,筛选各数据库建库至2023年成骨细胞铁死亡与激素性股骨头坏死及中草药干预调控研究相关的文章,最终纳入74篇文献进行综述分析。结果与结论:(1)成骨细胞铁死亡在激素性股骨头坏死发病中起重要作用。(2)成骨细胞铁死亡的发生受到多种机制通路调控,如细胞内铁超载引起铁死亡;细胞发生脂质过氧化损伤细胞膜引起铁死亡;细胞膜上胱氨酸/谷氨酸逆向转运蛋白通过影响谷胱甘肽水平和谷胱甘肽过氧化物酶4活性,从而诱导铁死亡;细胞内发生芬顿反应产生大量活性氧引起铁死亡等。(3)中药单体淫羊藿苷等、中药复方青娥丸等及中成药补肾活血颗粒等均可通过调控成骨细胞铁死亡的发生,有助于防治激素性股骨头坏死。(4)目前关于成骨细胞铁死亡相关机制尚不明确,继续深入探明两者的作用机制,有望为临床治疗激素性股骨头坏死提供新选择。

基于“成骨-成血管”理论探讨全身振动疗法治疗激素性股骨头坏死的效果及作用机制

目的:观察全身振动疗法(whole body vibration therapy,WBVT)治疗激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)的效果,并探讨其治疗SONFH的作用机制。方法:将50只SD大鼠随机分为空白组、模型组、WBVT组、Yoda1组、WBVT联合蜘蛛毒液肽(Grammostola spatulata mechanotoxin 4,GsMTx4)组。模型组、WBVT组、Yoda1组、WBVT联合GsMTx4组大鼠采用脂多糖联合甲泼尼龙琥珀酸钠构建SONFH模型。造模后,WBVT组使用WBVT干预,Yoda1组使用Piezo1蛋白激动剂Yoda1干预,WBVT联合GsMTx4组使用WBVT和Piezo1蛋白抑制剂GsMTx4干预。干预结束后,进行大鼠股骨头组织病理学观察(计算股骨头空骨陷窝率)、骨微结构观察,以及股骨头内Piezo1、骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)、Runt相关转录因子2(Runt-related transcription factor 2,Runx2)、低氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、分化簇31(cluster of differentiation 31,CD31)/内皮粘蛋白(endomucin,EMCN)蛋白表达量检测。结果:(1)大鼠股骨头组织病理学观察结果。空白组大鼠的股骨头内骨小梁致密且排列整齐。与空白组相比,模型组大鼠股骨头内的骨小梁较为稀疏,骨小梁细小、不连续,且排列紊乱。与模型组相比,WBVT组和Yoda1组大鼠的股骨头内骨小梁数量增多,排列较为整齐。与WBVT组相比,WBVT联合GsMTx4组大鼠的股骨头内骨小梁排列则较为紊乱。模型组、WBVT组、Yoda1组、WBVT联合GsMTx4组大鼠的股骨头空骨陷窝率均高于空白组(P=0.000,P=0.000,P=0.000,P=0.000),WBVT组、Yoda1组大鼠的股骨头空骨陷窝率均低于模型组(P=0.000,P=0.000),WBVT联合GsMTx4组大鼠的股骨头空骨陷窝率高于WBVT组(P=0.000)。(2)大鼠股骨头骨微结构观察结果。WBVT组和Yoda1组大鼠的股骨头骨体积分数、骨小梁厚度、骨小梁数量、骨小梁分离度与空白组的差异均无统计学意义(P=0.213,P=0.081,P=0.384,P=0.471;P=0.435,P=0.131,P=0.104,P=0.126)。模型组和WBVT联合GsMTx4组大鼠的股骨头骨体积分数、骨小梁厚度、骨小梁数量均低于空白组(P=0.000,P=0.000,P=0.000;P=0.000,P=0.000,P=0.000),骨小梁分离度均高于空白组(P=0.000,P=0.000)。WBVT组和Yoda1组大鼠的股骨头骨体积分数、骨小梁厚度、骨小梁数量均高于模型组(P=0.000,P=0.002,P=0.000;P=0.000,P=0.007,P=0.014),骨小梁分离度均低于模型组(P=0.000,P=0.000)。WBVT组大鼠的股骨头骨体积分数、骨小梁厚度、骨小梁数量、骨小梁分离度与Yoda1组的差异均无统计学意义(P=0.194,P=0.223,P=0.332,P=0.071)。WBVT联合GsMTx4组大鼠的股骨头骨体积分数、骨小梁厚度、骨小梁数量均低于WBVT组(P=0.002,P=0.021,P=0.000),骨小梁分离度高于WBVT组(P=0.000)。(3)大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量检测结果。WBVT组和Yoda1组大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量与空白组的差异均无统计学意义(P=0.061,P=0.122,P=0.773,P=0.814,P=0.991;P=0.112,P=0.071,P=0.955,P=0.749,P=0.915)。模型组和WBVT联合GsMTx4组大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量均低于空白组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000;P=0.000,P=0.000,P=0.000,P=0.000,P=0.000)。WBVT组和Yoda1组大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量均高于模型组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000;P=0.000,P=0.000,P=0.000,P=0.000,P=0.000)。WBVT组大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量与Yoda1组的差异均无统计学意义(P=0.962,P=0.179,P=0.214,P=0.990,P=0.975)。WBVT联合GsMTx4组大鼠股骨头内Piezo1、BMP2、Runx2、HIF-1α、VEGF蛋白表达量均低于WBVT组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000)。(4)大鼠股骨头内CD31/EMCN蛋白表达量检测结果。模型组和WBVT联合GsMTx4组大鼠股骨头内CD31/EMCN表达量均低于空白组(P=0.000,P=0.000)。WBVT组和Yoda1组大鼠股骨头内CD31/EMCN表达量与空白组的差异均无统计学意义(P=0.412,P=0.991)。WBVT组和Yoda1组大鼠股骨头内CD31/EMCN表达量均高于模型组(P=0.000,P=0.000)。WBVT联合GsMTx4组大鼠股骨头内CD31/EMCN表达量低于WBVT组(P=0.000)。结论:WBVT可以促进股骨头坏死组织修复,其作用机制可能与上调Piezo1蛋白的表达影响HIF-1α/VEGF轴,进而促进股骨头内H型血管的生成、改善股骨头血供有关。

Gubi Tongxiao granules regulate vasculization and apoptosis to intervene steroid-induced femoral head necrosis in rabbits

Objective:To investigate the effect of Gubi Tongxiao granules on vascular formation and cell apoptosis in the process of glucocorticoid-induced femoral head necrosis.Methods:Thirty experimental New Zealand rabbits were randomly divided into groups.Except the control group,the animal model of steroid-induced femoral head necrosis was established by lipopolysaccharide combined with glucocorticoids.After successful modeling,the experimental group was given Gubi Tongxiao granules intragastric treatment,the model and control group were given the same amount of normal saline intragastric treatment,once a day,for 8 weeks,the experimental animals were sacrificed,the bilateral femoral head of each group was taken out for hematoxylin-eosin staining,and protein CD34,CYR61 and VEGF were immunohistochemical staining localization.TUNEL staining was used to observe the apoptosis of femoral head cells.The expression levels of apoptosis proteins Bax and Bcl-2 were detected by WB.The expression of VEGF eNOs Bax and Bcl-2 genes was analyzed by RT-qPCR.Results:Gubi Tongxiao granules reduced the number of hollow bone lacunae and apoptotic positive cells(P<0.01),and up-regulated the expressions of CD34,CYR61 and VEGF in femoral head tissue.Compared with control group,the expression of pro-apoptotic protein Bax increased and the expression of anti-apoptotic protein Bcl-2,VEGF and eNOs mRAN decreased significantly in the model group(P<0.01),the opposite results were obtained in the experimental group after Gubi Tongxiao granule intervention(P<0.05).Conclusion:Gubi Tongxiao granules can improve the expression of angiogenic genes and regulate apoptosis-related proteins in the bone tissue of rabbit model with steroid-induced necrosis of femoral head,promote angiogenic differentiation and inhibit apoptosis,and thus achieve the effect of treating steroid-induced necrosis of femoral head.

HISTOMORPHOMETRIC STUDY OF SUBCHONDRAL BONE OF NECROTIC FEMORAL HEAD IN RABBITS INDUCED BY METHYLPREDNISOLONE COMBINED WITH LOW-DOSE LIPOPOLYSACCHARIDE

Objective To investigate the histomorphometric features of the necrotic femoral head of rabbits induced by methylprednisolone combined with lipopolysaccharide. Methods Thirty-two mole adult New Zealand white rabbits were used. Among them, 16 were injected with lipopolysaccharide and methylprednisolone （ osteonecrosis group） , and another 16 were sham-injected with saline （ control group）. Magnetic resonance （MR） imaging was taken for femoral heads of the rabbits in both groups at the end of 2, 4 and 6 weeks after the injection. All the rabbits were then killed 6 weeks later. The femoral heads of the rabbits were collected and processed for histological and histomorphometric analysis. Results Femoral head necrosis occurred in 14 rabbits of the osteonecrosis group which were confirmed by histological evaluation and MR imaging. Osteonecrotic femoral heads, compared to controls, were characterized by lower values of bone volume/tissue volume （ P 〈 0. 01 ）, tabecular thichness （ P 〈 0. 01 ） , osteoid surface/bone surface （ P 〈 0. 05 ）, mineralizing apposition rate （ P 〈 0 05 ）, and bone formation rate/bone surface （ P 〈0. 05 ）. However, tabecular separation and eroded surface/bone surface were higher in osteonecrotic femoral heads than in controls （ P 〈 0. 01 ）. Trabecular number and osteoclast surface/ bone surface did not differ significantly. Conclusion The results demonstrated that osteoporosis was apparent in osteonecrotic femoral heads induced by methylprednisolone and lipopolysaccharide, due mainly to trabecular thinning rather than reduction of trabecular number. This might be due to reduced bone formation combined with increased bone resorption.

Research Progress of Tonifying Kidney and Promoting Blood Circulation in the Treatment of Steroid-induced Necrosis of Femoral Head

Osteonecrosis of the femoral head is a common disability disease of the hip joint in China,and osteonecrosis of the femoral head caused by hormone factors is the most common,which is related to the gradual increase in the utilization rate of glucocorticoids in recent years.It is a refractory disease in orthopaedics with a poor prognosis.For this disease,the treatment of traditional Chinese medicine has certain advantages.In view of this,the author reads,analyzes and summarizes its materials by searching China Journal full-text Database and Wanfang Database.This paper reviews the mechanism and clinical research progress of tonifying kidney and activating blood circulation in the prevention and treatment of steroid-induced osteonecrosis of the femoral head,hoping to provide help for the clinical treatment of hormone-related osteonecrosis of the femoral head.

主体间性视域下大学生班主任工作的三个维度

大学生班主任工作是高校人才培养工作的重要组成部分。长期以来,高校大学生班主任工作中存在主体意识、主体间性认知的偏差,导致班主任工作理论基础缺失并面临实践困境。在主体间性视域下,文章以厘清多主体的存在及多元主体间关系为基础,分析大学生班主任的根本属性、基本属性和岗位属性,重构大学生班主任工作中的师师关系、师生关系、生生关系,设计大学生班主任工作的实践路径,这对于明确大学生班主任工作的思维逻辑、价值追求和实践方法,促进新时期大学生班主任育人目标的实现具有重要意义。

葛根素通过AMPK-mTOR-ULK1通路激活自噬改善大鼠激素性股骨头坏死的研究

目的探讨葛根素通过AMPK-mTOR-ULK1信号通路对激素性股骨头坏死早期血管内皮细胞自噬的干预作用。方法将48只雄性SD大鼠按随机数表法分成4组(每组12只)。对照组:单纯肌肉注射生理盐水,2 mL/次,共3次,间隔24 h。模型组:肌肉注射甲基强的松龙,20 mg/kg,共3次,间隔24 h。葛根素干预组:同模型组方法造模,在最后1次臀肌注射完甲基强的松龙后给予葛根素100 mg/(kg·d)腹腔注射。雷帕霉素干预组:同模型组方法造模,在最后1次臀肌注射完甲基强的松龙后同时给予雷帕霉素[1.2 mg/(kg·d),自噬特异性激动剂]腹腔注射。造模后2周、4周后处死实验动物,取出双侧后肢的股骨头及主要供血血管。HE染色光镜下观察股骨头坏死情况;利用免疫组化法和RT-PCR方法检测Beclin1、LC3、P62、AMPK、mTOR和ULK1的mRNA与蛋白的表达;采用ELISA法检测血管内皮损伤标记物6-keto-PGF1a的表达。结果与对照组比较,模型组大鼠骨细胞坏死加重,血管内皮细胞6-keto-PGF1a的表达增加,差异有统计学意义(P<0.05),血管内皮细胞中自噬相关蛋白AMPK、ULK1、Beclin1、LC3表达降低,差异有统计学意义(P<0.05),mTOR、P62表达增高,差异有统计学意义(P<0.05);与模型组比较,葛根素干预组大鼠的骨坏死程度明显改善,6-keto-PGF1a的表达降低,差异有统计学意义(P<0.05),自噬相关蛋白AMPK、ULK1、Beclin1、LC3表达增高,差异有统计学意义(P<0.05),mTOR、P62表达降低,差异有统计学意义(P<0.05)。结论葛根素可能通过AMPK-mTOR-ULK1信号通路激活血管内皮细胞的自噬,从而减轻激素所造成的股骨头坏死。

肠道菌群变化对大鼠激素性股骨头坏死造模效果的影响

目的探究激素性股骨头坏死(GA-ONFH)模型大鼠的肠道菌群变化及其对造模效果的影响。方法40只雄性SD大鼠随机分为空白对照组、抗生素组(ABX组)、激素性股骨头坏死模型组(GA-ONFH模型组)、抗生素+激素性股骨头坏死模型组(ABX+GA-ONFH组),每组10只。ABX组和ABX+GA-ONFH组持续给予抗生素溶液洗脱肠道微生物1周。1周后GA-ONFH模型组和ABX+GA-ONFH组给予脂多糖(LPS)+地塞米松腹腔注射建立早期GA-ONFH模型,空白对照组和ABX组腹腔注射等量生理盐水,连续注射6周。收集大鼠粪便和股骨头样本,基于16S rDNA扩增子测序分析大鼠肠道菌群的构成及丰度,使用micro-CT和HE染色观察早期GA-ONFH造模效果,评估肠道菌群对GA-ONFH造模效果的影响。结果与空白对照组比较,GA-ONFH模型组大鼠肠道菌群丰度发生改变,其中拟杆菌门丰度明显下调,而厚壁菌门丰度明显上调,差异有统计学意义(P<0.05);在科水平上,普雷沃菌科、梭状芽胞杆菌科、消化球菌科、理研菌科和克里斯滕森菌科的丰度明显下调,而葡萄球菌科的丰度明显上调,差异有统计学意义(P<0.05);在属水平上,另枝菌属、UCG-005、梭状芽胞杆菌属、拟普雷沃菌属和普雷沃菌属的丰度明显下调,而葡萄球菌属和Frisingicoccus的丰度明显上调,差异有统计学意义(P<0.05)。Micro-CT断层扫描显示,GA-ONFH模型组和ABX+GAONFH组均出现软骨下骨小梁硬化;HE染色显示,ABX+GA-ONFH组较GA-ONFH模型组更早出现骨小梁断裂,表明抗生素洗脱肠道微生物后,大鼠GA-ONFH造模效果更加显著。结论GA-ONFH模型大鼠的肠道菌群发生了明显变化,且该变化可能影响早期GA-ONFH的发生。

股骨头坏死发病机制中非编码RNA的靶标性

背景:目前非编码RNA已被证实在多种骨科疾病中发挥重要作用,因此,非编码RNA可能为了解该疾病的病因、发病机制、治疗方法带来新的思路。目的:探索早期诊断、治疗股骨头坏死的方法和思路,综述非编码RNA在股骨头坏死中的研究进展,并提出目前所存在的问题及解决方法。方法:以“股骨头坏死,非编码RNA,miRNA,lncRNA,circRNA”为检索词检索中国知网、万方、维普等数据库,以“osteonecrosis of the femoral head,non-coding RNAs,miRNA,lncRNA,circRNA”为检索词检索PubMed、Medline等数据库,检索时限均为2000年1月至2021年11月。最终纳入67篇文献。结果与结论:(1)非编码RNA的种类非常丰富,大约占所有转录产物的98%,同时占所有RNA的90%左右,根据其功能可分为管家非编码RNA(如tRNA、rRNA、snoRNA)以及调节非编码RNA(如miRNA、lncRNA、circRNA、siRNA);(2)目前miRNAs与激素型股骨头坏死的研究逐渐成为热点,而外泌体lncRNAs、circRNAs与股骨头坏死的相关研究尚未见到报道;(3)多数非编码RNA在发生激素型股骨头坏死时表达下调,大部分非编码RNA通过负向调控其相关靶基因,以促进骨髓间充质干细胞增殖、成骨分化,促进成骨标记物的表达,也可以促进血管内皮细胞生成,从而抑制股骨头缺血性坏死的发生及进展;(4)外泌体携带的非编码RNA也可以在股骨头坏死中发生作用,但目前文献量较少,非编码RNA是否通过外泌体携带所产生的作用目前暂不清楚,在未来的研究中可逐渐完善;(5)利用非编码RNA类似物和抑制剂可以有效抑制非创伤性股骨头坏死的发生发展,可以为非创伤性股骨头坏死的早期诊断和治疗提供新的策略。

知识增强的自然语言生成研究综述

自然语言生成(Natural Language Generation,NLG)任务是自然语言处理(Natural Languge Processing,NLP)任务中的一个子类,并且是一项具有挑战性的任务。随着深度学习在自然语言处理中的大量应用,其已经变成自然语言生成中处理各种任务的主要方法。自然语言生成任务中主要有问答任务、生成摘要任务、生成评论任务、机器翻译任务、生成式对话任务等。传统的生成模型依赖输入文本,基于有限的知识生成文本。为解决这个问题,引入了知识增强的方法。首先介绍了自然语言生成的研究背景和重要模型,然后针对自然语言处理归纳介绍了提高模型性能的方法,以及基于内部知识(如提取关键词增强生成、围绕主题词等)和外部知识(如借助外部知识图谱增强生成)集成到文本生成过程中的方法和架构。最后,通过分析生成任务面临的一些问题,讨论了未来的挑战和研究方向。

骨痹通消颗粒调控Wnt/PI3K⁃AKT信号通路干预兔激素性股骨头坏死

目的:探讨骨痹通消颗粒通过调控Wnt/PI3K⁃AKT信号通路对兔激素性股骨头坏死的治疗作用。方法:将30只清洁级新西兰兔随机分为对照组、模型组和实验组,除对照组外,其余均采用异体血清联合糖皮质激素建立模拟激素性股骨头坏死动物模型。造模成功后,模型组与对照组给予等量生理盐水,实验组按照人与兔的等效剂量给予骨痹通消颗粒浓缩液,1天1次,持续8周,经腹腔麻醉后体外心脏采血并采用免疫酶联吸附法检测血清中ALP、TGF⁃β1水平;处死实验动物,取出各组兔双侧股骨头进行苏木精⁃伊红染色,TUNEL染色法观察股骨头区细胞凋亡情况;RT⁃qPCR分析Bax、Bcl⁃2基因表达情况,Western blot检测通路蛋白Wnt、β⁃catenin、GSK⁃3β、AKT和p⁃AKT等表达水平。结果:与对照组相比,模型组兔骨组织中ALP和TGF⁃β1表达水平显著下降(P<0.05),骨小梁稀疏变细,空骨陷窝数和凋亡阳性细胞数明显增多(P<0.05),骨组织中促凋亡基因Bax表达升高,抗凋亡基因Bcl⁃2表达则受到抑制(P<0.05);经骨痹通消颗粒干预治疗后,实验组兔骨组织空骨陷窝情况和细胞凋亡情况均得到有效改善(P<0.05)。此外,与模型组对比,骨痹通消颗粒能够上调ALP和TGF⁃β1表达量(P<0.05),同时促进通路蛋白Wnt、β⁃catenin表达,提高AKT磷酸化水平,下调GSK⁃3β的表达(P<0.05)。结论:糖皮质激素环境下,成骨细胞发育功能障碍,细胞凋亡增多。骨痹通消颗粒能够调控Wnt/PI3K⁃AKT两条信号通路,促进成骨细胞发育并抑制细胞凋亡,有效维持骨代谢平衡,延长股骨头塌陷时间,进一步治疗股骨头坏死。