女性致密性骨炎血清骨转换生化标志物水平变化研究

背景 致密性骨炎（OCI）和其他疾病有时难以鉴别，探讨血清骨转换生化标志物可为OCI的鉴别诊断提供依据。 目的 探索女性OCI患者的血清骨转换生化标志物的水平变化及临床意义。 方法 回顾性选取2013年6月至2022年2月在北京积水潭医院门诊及住院诊断为OCI的61例女性患者作为观察组，年龄15~50岁，平均（33.8±6.6）岁，病程2周~15年。选择同期61例女性体检健康者作为对照组，年龄15~48岁，平均（35.6±7.6）岁。比较两组一般临床资料和血清骨转换生化标志物水平，并对血清骨转换生化标志物与病情相关指标进行相关性分析。 结果 观察组血清白蛋白（45.4±2.9）g/L低于对照组（46.5±2.8）g/L（t=2.190，P<0.05）。血清骨转换生化标志物比较结果显示，观察组血清1型胶原羧基末端肽β特殊序列（β-CTX）〔0.28（0.23，0.37）μg/L〕、N-端骨钙素（OC）〔13.1（11.2，16.2）μg/L〕、25-羟维生素D3〔25-（OH）VD3〕〔（14.1±5.1）μg/L〕低于对照组〔0.36（0.29，0.48）μg/L，15.6（13.7，17.3）μg/L，（17.5±6.6）μg/L〕（Z=-2.983、-3.255，t=3.081，P<0.05）。长病程亚组OC水平〔14.6（12.4，18.5）μg/L〕高于短病程亚组〔11.7（10.2，14.0）μg/L〕（Z=-2.407，P<0.05）。多孕亚组β-CTX〔0.25（0.22，0.32）μg/L〕、OC水平〔12.2（10.3，15.0）μg/L〕低于非多孕亚组〔0.33（0.26，0.44）μg/L、13.4（12.0，18.8）μg/L〕（Z=-2.486、-1.897，P<0.05）。相关性分析显示，观察组血清1型前胶原氨基端延长肽（tP1NP）与妊娠次数、生产次数均呈负相关（rs=-0.276、-0.298，P<0.05），OC与体质指数（BMI）、视觉模拟评分法（VAS）评分、妊娠次数均呈负相关（rs=-0.284、-0.374、-0.360，P<0.05），25-（OH）VD3水平与BMI呈正相关（rs=0.275，P<0.05）。 结论 女性OCI患者血清OC、β-CTX水平明显降低，可为鉴别其他疾病提供依据；血清OC水平可以反映OCI患者的严重程度，同时OC水平与患者妊娠次数相关；tP1NP与妊娠次数、生产次数相关。     

Hepatotoxicity of cantharidin is associated with the altered bile acid metabolism

Cantharidin (CTD) is an effective antitumor agent. However, it exhibits significant hepatotoxicity, the mechanism of which remains unclear. In this study, biochemical and histopathological analyses complemented with ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS)-based targeted metabolomic analysis of bile acids (BAs) were employed to investigate CTD-induced hepatotoxicity in rats. Sixteen male and female Sprague–Dawley rats were randomly divided into two groups: control and CTD (1.0 mg/kg) groups. Serum and liver samples were collected after 28 days of intervention. Biochemical, histopathological, and BA metabolomic analyses were performed for all samples. Further, the key biomarkers of CTD-induced hepatotoxicity were identified via multivariate and metabolic pathway analyses. In addition, metabolite–gene–enzyme network and Kyoto Encyclopedia of Genes and Genomes pathway analyses were used to identify the signaling pathways related to CTD-induced hepatotoxicity. The results revealed significantly increased levels of biochemical indices (alanine aminotransferase, aspartate aminotransferase, and total bile acid). Histopathological analysis revealed that the hepatocytes were damaged. Further, 20 endogenous BAs were quantitated via UHPLC-MS/MS, and multivariate and metabolic pathway analyses of BAs revealed that hyocholic acid, cholic acid, and chenodeoxycholic acid were the key biomarkers of CTD-induced hepatotoxicity. Meanwhile, primary and secondary BA biosynthesis and taurine and hypotaurine metabolism were found to be associated with the mechanism by which CTD induced hepatotoxicity in rats. This study provides useful insights for research on the mechanism of CTD-induced hepatotoxicity.     

HPLC-MS/MS测定连翘中连翘酯苷A、阿魏酸、槲皮素、松脂醇β-D葡萄糖苷、连翘苷的含量

[目的]建立采用HPLC-MS/MS测定连翘中连翘酯苷A、阿魏酸、槲皮素、松脂醇β-D葡萄糖苷、连翘苷的含量的方法,为其开发应用提供方法参考。[方法]色谱柱选用Phenomenex Luna C18柱(150 mm×2. 0 mm,5μm);流动相A:0. 1%甲酸溶液,流动相B:100%乙腈;流速为0. 7 m L/min;进样量为12μL;洗脱梯度设定为:0～5 min,10%～55%B; 5～11 min,55%～100%B。质谱选择负离子扫描,离子源为电喷雾式(ESI)。采用多反应监测的检测模式,喷雾电压:-4. 7 k V,雾化气压0. 5 MPa,雾化温度620℃,氮气压力0. 65 MPa。[结果]本次研究所检测的连翘在可测定浓度的范围内5种成分的峰面积与浓度都表现出较好的线性关系,加样后的回收率在97. 8%～99. 5%之间。[结论]采用HPLC-MS/MS测定连翘中5中成分的含量效果较好,灵敏度高、结果可靠,为连翘的开发应用以及质量控制提供了参考。     

基于多元统计分析的银杏叶提取物中有机酸类成分含量研究

目的建立HPLC法测定银杏叶提取物中9种小分子有机酸类成分莽草酸、儿茶素、表儿茶素、没食子酸、原儿茶酸、6-羟基犬尿喹啉酸、对羟基苯甲酸、对羟基肉桂酸、咖啡酸含量的方法,并结合多元统计分析方法比较不同厂家生产的银杏叶提取物间质量差异。方法采用Inertsil ODS-3 C_(18)(250 mm×4.6 mm,5μm)色谱柱;以乙腈-0.4%磷酸水溶液为流动相,梯度洗脱,波长切换测定(220 nm检测莽草酸、儿茶素、表儿茶素,254 nm检测没食子酸、原儿茶酸、6-羟基犬尿喹啉酸、对羟基苯甲酸,310 nm检测对羟基肉桂酸、咖啡酸),柱温40℃。结果不同厂家生产的银杏叶提取物中有机酸总含量有差异,厂家内部样品中各有机酸含量同样存在差异。经聚类分析、主成分分析、相关与回归分析等多元统计方法分析,筛选出儿茶素、没食子酸、原儿茶酸及6-羟基犬尿喹啉酸为体现样品质量差异的特征成分,同时成分间具有内在相关性。结论建立的方法操作简便、重复性好、结果可靠,可用于银杏叶提取物中有机酸类成分的质量评价。筛选出的儿茶素、没食子酸、原儿茶酸及6-羟基犬尿喹啉酸是体现质量差异的特征成分,又是具有内在关联的共性成分,为提升银杏叶提取物整体质量控制能力提供了依据,同时表明银杏叶提取物的提取工艺并不统一,企业内部工艺的稳定性也有待提高。     

Effect of a Device-Free Compressed Shell Fixation Method on Hepatic Respiratory Movement: Analysis for Respiratory Amplitude of the Liver and Internal Motions of a Fiducial Marker

Purpose

Suppression of respiratory movement of the liver would be desirable for high-precision radiation therapy for liver tumors. We aimed to investigate the effect of our original device-free compressed shell fixation method and breathing instruction on suppression of respiratory movement. The characteristics of liver motion based on the movement of a fiducial marker were also analyzed.

Dysregulation of sphingosine 1 phosphate receptor-1 (S1P1) signaling and regulatory lymphocyte-dependent immunosuppression in a model of post-fingolimod MS rebound

Fingolimod affords protection from MS by sequestering lymphocytes in secondary lymphoid organs via down regulation of their sphingosine 1 phosphate receptor (S1P1). Unexpectedly, accumulating evidence indicates that patients who discontinue fingolimod treatment may be at risk of rehearsal of magnetic resonance (MR) and clinical disease activity, sometimes featuring dramatic rebound.We therefore developed in vivo and in vitro models of post-fingolimod MS rebound to unravel its cellular and molecular mechanisms. The impact of fingolimod withdrawal on T regulatory lymphocytes was also investigated by means of cytofluorimetric analysis and antigen-specific lymphocyte proliferation assays. We show that mice with relapsing-remitting experimental autoimmune encephalomyelitis (EAE) undergo extremely severe, chronic disease rebound upon discontinuation of fingolimod. Remarkably, rebound is preceded by a burst of S1P1 overexpression in lymph node-entrapped lymphocytes that correlates with subsequent massive lymphocyte egress and widespread CNS immune infiltration. Also, consistent with the ability of S1P1 to counteract polarization and function of T regulatory lymphocytes their number and suppression of effector T cells is reduced by fingolimod suspension. Data disclose the first pathogenic mechanisms of post-fingolimod rebound that may be targeted for therapeutic intervention.