早产儿急性呼吸窘迫综合征血清1,25-(OH)2D3、PGRN、SIRT1、CTRP3水平与炎症反应和预后的关系研究

摘要 目的：探讨不同病情严重程度早产儿急性呼吸窘迫综合征（ARDS）血清1,25-二羟维生素D₃（1,25-(OH) ₂D₃）、颗粒体蛋白前体（PGRN）、沉默信息调节因子2相关酶1（SIRT1）、C1q/肿瘤坏死因子相关蛋白3（CTRP3）的变化，分析其与炎症反应和预后的关系。方法：选择2018年10月至2019年12月安徽省妇幼保健院收治的ARDS早产儿100例作为ARDS组，另选取同期在我院出生的健康新生儿60例作为对照组。根据《"新生儿急性呼吸窘迫综合征"蒙特勒标准（2017年版）》的病情判定标准将ARDS早产儿分为轻度组（n=40）、中度组（n=32）、重度组（n=28），比较不同病情严重程度ARDS早产儿血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3、炎症反应指标肿瘤坏死子因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）水平变化，采用Pearson法分析ARDS早产儿血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3与炎症反应指标的相关性。另根据ARDS组患儿预后情况分为预后良好组（n=65）和预后不良组（n=35），采用单因素和多因素Logistic回归分析影响ARDS早产儿预后不良的危险因素。结果：ARDS组血清1,25-(OH) ₂D₃、SIRT1、CTRP3、PGRN水平明显低于对照组（P<0.05），ARDS组血清TNF-α、IL-6、IL-1β水平明显高于对照组（P<0.05）。不同病情严重程度ARDS早产儿血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3、炎症反应指标比较差异有统计学意义（P<0.05）。ARDS早产儿血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3水平与TNF-α、IL-6、IL-1β水平呈负相关（P<0.05）。多因素Logistic回归分析结果显示，低出生体重、肺表面活性物质（PS）使用次数≥3次、出现低白蛋白血症是影响ARDS早产儿预后不良的危险因素（P<0.05），血清1,25-(OH)2D3（较高）、PGRN（较高）、SIRT1（较高）、CTRP3（较高）是ARDS早产儿预后不良的保护因素（P<0.05）。结论：血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3可能参与ARDS早产儿的炎症反应过程，与ARDS早产儿的病情进展及预后密切相关，检测血清1,25-(OH) ₂D₃、PGRN、SIRT1、CTRP3有助于评估ARDS早产儿的预后。     

不明原因复发性流产再次妊娠患者血清1,25(OH)2D3、sTim-3与Th17/Treg免疫失衡和妊娠结局的关系

摘要 目的：探讨不明原因复发性流产（URSA）再次妊娠患者血清1,25-二羟维生素D3[1,25（OH)）₂D₃]、可溶性T细胞免疫球蛋白黏蛋白分子3（sTim-3）与辅助性T细胞17（Th17）/调节性T细胞（Treg）免疫失衡和妊娠结局的关系。方法：选择于湖南省妇幼保健院2020年1月~2022年1月就诊的62例URSA再次妊娠患者作为研究组,另选择同期进行孕检的正常早孕妇女30例作为对照组。比较两组孕早期血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平、Th17/Treg比值。Pearson法分析URSA再次妊娠患者血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值平的相关性。根据URSA再次妊娠患者妊娠结局的不同分为妊娠成功分娩组和妊娠再次流产组,比较两组孕早期血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值。受试者工作特征（ROC）曲线分析血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值对妊娠结局的预测价值。结果：研究组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于对照组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于对照组（P<0.05）。Pearson相关分析显示,URSA再次妊娠患者血清1,25(OH) ₂D₃与血清sTim-3、外周血Th17细胞水平、Th17/Treg比值呈负相关,与Treg细胞水平呈正相关（P<0.05）;血清sTim-3与外周血Treg细胞水平呈负相关,与Th17细胞水平、Th17/Treg比值呈正相关（P<0.05）。妊娠再次流产组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于妊娠成功分娩组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于妊娠成功分娩组（P<0.05）。ROC曲线分析显示,血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平及Th17/Treg比值均可预测URSA再次妊娠患者妊娠再次流产的发生风险,且上述指标联合检测的预测效能更高。结论：血清1,25(OH) ₂D₃水平异常降低、sTim-3水平异常升高可导致Th17/Treg免疫失衡,导致URSA再次妊娠患者再次发生流产。上述指标联合检测对URSA再次妊娠患者妊娠再次流产的预测效能更高。     

龈沟液Chemerin、APN、25(OH)D3与伴2型糖尿病的慢性牙周炎患者牙周指标和Th17/Treg失衡的关系

摘要 目的：探讨龈沟液趋化素（Chemerin）、脂联素（APN）、25-羟维生素D3 [25(OH)D3]与伴2型糖尿病（T2DM）的慢性牙周炎（CP）患者牙周指标和Th17/Treg失衡的关系。方法：选择2021年1月至2022年1月我院口腔科门诊接诊的125例伴T2DM的CP患者,根据CP病情严重程度分为轻度组（42例）、中度组（53例）和重度组（30例）。检测龈沟液中Chemerin、APN、25(OH)D₃水平以及外周血中Th17细胞占比、Treg细胞占比,计算Th17/Treg比值,记录出血指数（SBI）、菌斑指数（PLI）、附着丧失（AL）、牙周袋探诊深度（PD）。分析龈沟液中Chemerin、APN、25(OH)D₃与牙周指标、外周血Th17/Treg的相关性。结果：重度组龈沟液 Chemerin水平和外周血中Th17细胞占比、Th17/Treg比值、PLI、SBI、AL、PD高于中度组和轻度组（P＜0.05）,龈沟液APN、25(OH)D₃水平,外周血Treg细胞占比低于中度组和轻度组（P＜0.05）。龈沟液 Chemerin与PLI、SBI、AL、PD、外周血Th17细胞占比、Th17/Treg比值呈正相关（P＜0.05）,与Treg细胞占比呈负相关（P＜0.05）;龈沟液APN、25(OH)D₃与PLI、SBI、AL、PD、外周血中Th17细胞占比、Th17/Treg比值呈负相关（P＜0.05）,与Treg细胞占比呈正相关（P＜0.05）。结论：伴T2DM 的CP患者龈沟液中Chemerin水平增高,APN、25(OH)D₃水平降低,且与牙周指标和Th17/Treg失衡有关。     

体型肥胖2型糖尿病患者血清胆红素、胆汁酸、25(OH)D3水平及与胰岛素抵抗的关系研究

摘要 目的：探讨体型肥胖2型糖尿病（T2DM）患者血清胆红素、胆汁酸（BA）、25羟维生素D3[25(OH)D₃]水平及其与胰岛素抵抗（IR）的关系。方法：选取2018年1月-2019年12月我院就诊的240例T2DM患者,根据体重指数（BMI）将患者分为T2DM-N组84例,T2DM-OW组92例,T2DM-OB组64例。分别测定总胆红素（TBIL）、间接胆红素（IBIL）、直接胆红素（DBIL）、胆汁酸（BA）、25(OH)D₃等指标。以胰岛素抵抗指数（HOMA-IR）为因变量,行多元线性回归,分析IR的危险因素。结果：T2DM-OB组BMI、BA、甘油三酯（TG）、空腹血糖（FPG）、餐后2小时血糖（2hPG）、糖化血红蛋白（HbA1c）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）、C反应蛋白（CRP）高于T2DM-N组,DBIL、25(OH)D₃低于T2DM-N组,差异均有统计学意义（P<0.05）。Pearson相关性分析显示,HOMA-IR与BMI、BA呈正相关（P<0.05),与DBIL、25(OH)D₃呈负相关（P<0.05);多元线性回归分析显示BMI升高,25(OH)D₃、DBIL下降为IR的危险因素（P<0.05）。结论：肥胖的T2DM患者存在IR及血脂紊乱,心血管疾病患病风险增加,BA水平升高,DBIL、25(OH)D₃水平下降,机体慢性低度炎症水平升高,除了BMI,25(OH)D₃、DBIL水平下降亦有可能是体型肥胖T2DM患者IR的重要危险因素。     

肺炎支原体肺炎合并喘息患儿血清25（OH）D3水平与肺功能的相关性及其影响因素分析

摘要 目的：探讨肺炎支原体肺炎（MPP）患儿合并喘息的影响因素,分析血清25-羟维生素D3[25(OH)D-3]水平与MPP合并喘息患儿肺功能的关系。方法：选择2017年1月至2020年1月我院收治的90例MPP患儿,根据是否合并喘息将其分为MPP合并喘息组（39例）和MPP未合并喘息组（51例）。检测血清25(OH)D-3水平以及肺功能[最大呼气流速（PEF）、PEF占预计值百分比(PEF% pred)、第一秒用力呼吸容积（FEV₁）、用力肺活量（FVC）、FEV₁/FVC比值、FEV₁占预计值百分比（FEV1%pred）],Pearson相关性分析25(OH)D-3与MPP合并喘息患儿肺功能的关系,单因素及多因素Logistic回归分析影响MPP合并喘息的危险因素。结果：MPP合并喘息组血清25(OH)D-3水平、PEF、PEF% pred、FEV₁/FVC比值、FEV1%pred低于MPP未合并喘息组（P＜0.05）,Pearson相关性分析显示,MPP合并喘息组患儿血清25(OH)D-3水平与PEF、PEF% pred、FEV₁/FVC比值、FEV₁%pred均呈正相关（r=0.519、0.612、0.571、0.593,P＜0.05）。单因素分析显示,MPP合并喘息组年龄低于MPP未合并喘息组（P＜0.05）,病程长于MPP未合并喘息组（P＜0.05）,肺部啰音比例、嗜酸性粒细胞计数、MP-IgM抗体滴度高于MPP未合并喘息组（P＜0.05）。多因素Logistic回归分析结果显示低龄、肺部啰音、嗜酸性粒细胞计数增高、MP-IgM抗体滴度增加、25(OH)D-3减少是MPP合并喘息的危险因素（P＜0.05）。结论：MPP合并喘息患儿25(OH)D-3水平较低,低龄、肺部啰音、嗜酸性粒细胞计数增高、MP-IgM抗体滴度增加、25(OH)D-3缺乏为MPP合并喘息的危险因素,25(OH)D-3缺乏与MPP合并喘息患儿肺功能下降有关。     

中重度支气管哮喘急性发作期患儿Fe NO表达水平与肺功能的相关性研究

摘要 目的：探究中重度支气管哮喘急性发作期患儿呼出气一氧化氮（Fe NO）表达水平与肺功能的相关性。方法：选择2016年3月-2019年3月来我院就诊的中重度支气管哮喘急性发作期患儿69例为观察组,其中,中度支气管哮喘急性发作期患儿58例,重度支气管哮喘急性发作期患儿11例;另选取同期来我院体检的69例正常健康儿童作为对照组,对比观察组中中度、重度支气管哮喘急性发作期患儿Fe NO表达水平、用力肺活量（forced vital capacity,FVC）、最大呼气流量占预计值百分比（Maximum expiratory flow as a percentage of expected value,PEF%）、第一秒用力呼气容积占预计值的百分比（Forced expiratory volume as a percentage of expected value in the first second,FEV1%）与对照组健康儿童的差异,并对观察组患儿Fe NO表达水平与肺功能的相关性进行分析。结果：观察组患儿的Fe NO表达水平均高于对照组,且重度组患儿的Fe NO表达水平明显高于中度组（P＜0.05）;观察组患儿的PEF%、FEV₁%、FVC水平均高于对照组,且重度组患儿的PEF%、FEV₁%、FVC水平均高于中度组（P＜0.05）;观察组患儿Fe NO表达水平与FVC、PEF%、FEV₁%指标均呈负相关关系（r=-0.503、-0.551、-0.532,P均＜0.05）。结论：中重度支气管哮喘急性发作期患儿Fe NO表达水平与肺功能成负相关,可通过监测 Fe NO 水平间接判断炎症程度。     

血清脑钠肽、25-羟维生素D、半乳糖凝集素-3与妊娠期高血压疾病患者左心室舒张功能的相关性及对妊娠结局的影响

摘要 目的：探讨妊娠期高血压疾病（HDP）患者血清脑钠肽（BNP）、25-羟维生素D[25(OH)D]、半乳糖凝集素-3 （Gal-3）与左心室舒张功能的相关性,并分析其对患者妊娠结局的影响。方法：选择2019年6月至2021年12月我院收治的HDP患者114例纳入研究组,其中妊娠期高血压组39例,轻度子痫前期组44例,重度子痫前期组31例,另选择同期在我院产检的健康孕妇30例作为对照组。比较各组血清BNP、25(OH)D、Gal-3及左心室舒张功能,分析患者血清BNP、25(OH)D、Gal-3与左心室舒张功能的相关性。根据妊娠结局将患者分为妊娠结局良好组65例、妊娠结局不良组49例,收集两组患者临床资料,应用单因素分析和多因素Logistic回归分析 HDP患者妊娠结局不良的影响因素。结果：各组血清BNP、25(OH)D、Gal-3水平比较差异有统计学意义（P<0.05）,HDP患者随疾病严重程度的升高血清BNP、Gal-3水平逐渐升高,25(OH)D水平逐渐降低。各组左心室舒张早期充盈峰速度（E峰）、左心室舒张晚期充盈峰速度（A峰）、E/A比较差异有统计学意义（P<0.05）。Pearson相关分析显示,HDP患者血清BNP、Gal-3水平与E峰、E/A呈负相关（P<0.05）,与A峰呈正相关（P<0.05）,血清25(OH)D水平与E峰、E/A呈正相关（P<0.05）,与A峰呈负相关（P<0.05）。多因素Logistic回归分析显示：收缩压≥158.02 mmHg、舒张压≥104.32 mmHg、血清BNP≥406.42 ng/L、血清25(OH)D≤14.94 μg/L、血清Gal-3≥16.17 ng/mL是HDP妊娠结局不良的危险因素（P<0.05）。结论：HDP患者血清BNP、Gal-3水平异常升高、25(OH)D水平异常降低均为HDP患者妊娠结局不良的危险因素,三者水平与患者左心室舒张功能相关,检测三者水平有助于对患者妊娠结局的临床判断。     

支气管哮喘患儿血清硫化氢、嗜酸性粒细胞趋化因子水平与炎症因子及肺功能的关系研究

摘要 目的：探讨支气管哮喘患儿血清硫化氢（H2S）、嗜酸性粒细胞趋化因子（Eotaxin）水平与炎症因子及肺功能的关系。方法：选取我院收治的95例支气管哮喘患儿,按照《儿童支气管哮喘诊断及防治指南(2016年版)》将支气管哮喘患儿分为急性发作期组43例和缓解期组52例。检测血清中H2S 水平;检测Eotaxin、肿瘤坏死因子-α（TNF-α）,白介素6（IL-6）和白介素13（IL-13）的水平;对患儿进行肺功能测定,采用Pearson检验分析血清H₂S、Eotaxin水平与血清炎症因子及肺功能之间的相关性。ROC曲线分析血清H2S、Eotaxin对支气管哮喘患儿病情的预测价值。结果：急性发作期组患儿血清H₂S水平、第一秒用力呼气容积占预计值百分比（FEV₁%pred）、最大呼气流量占预计值百分比（PEF%pred）和FEV₁/用力肺活量（FVC）低于缓解期组患儿,Eotaxin、TNF-α、IL-6和IL-13水平高于缓解期组患儿（P<0.05）;支气管哮喘患儿血清H₂S水平与TNF-α、IL-6和IL-13水平均呈负相关（P<0.05）,与FEV₁%pred、FEV₁/FVC和PEF%pred均呈正相关（P<0.05）;支气管哮喘患儿血清Eotaxin水平与TNF-α、IL-6和IL-13水平均呈正相关（P<0.05）,与FEV₁%pred、FEV₁/FVC和PEF%pred均呈负相关（P<0.05）。血清H₂S预测支气管哮喘患儿病情的曲线下面积为0.854,灵敏度和特异度分别为77.89%和81.05%,Youden指数为0.5894;血清Eotaxin预测支气管哮喘患儿病情的曲线下面积为0.924,灵敏度和特异度分别为92.71%和84.38%,Youden指数为0.7709。结论：支气管哮喘急性发作期患儿与缓解期患儿相比,其血清H2S水平较低,Eotaxin水平较高,血清H₂S和Eotaxin水平与机体炎症反应及肺功能密切相关,有助于辅助评估支气管哮喘患儿的病情。     

miR-1165-3p、miR-145水平在支气管哮喘患者中的表达及其临床意义

摘要 目的：探讨微小RNA（MicroRNA,miR）-1165-3p、miR-145水平在支气管哮喘患者中的表达及其临床意义。方法：收集2021年1月-2022年3月中国人民解放军总医院第六医学中心62例支气管哮喘患者作为研究组,其中轻度急性发作27例,中度急性发作22例,重度急性发作13例。另收集同时期、同年龄段30例健康体检者作为对照组。采用实时荧光定量PCR（RT-PCR）检测各组血清miR-1165-3p、miR-145表达水平。采用Spearman相关分析不同程度支气管哮喘患者与血清miR-1165-3p、miR-145之间的相关性。通过受试者工作特征（ROC）分析血清miR-1165-3p、miR-145表达水平对不同程度支气管哮喘的诊断效能。结果：与对照组相比,研究组中白细胞介素-6（IL-6）、嗜酸性粒细胞、总免疫球蛋白E（IgE）水平显著升高,第1秒用力呼气容积（FEV₁）占预测值百分比（FEV₁%）则显著降低,差异均有统计学意义（P＜0.05）。不同严重程度支气管哮喘患者（轻度、中度、重度）血清miR-1165-3p、miR-145表达水平均高于健康对照组,支气管哮喘越严重,其表达水平越高,且组间、组内比较差异均有统计学意义（P＜0.05）。Spearman相关分析显示,miR-1165-3p、miR-145、IL-6表达水平与哮喘严重程度呈正相关（P＜0.05）,FEV₁%与哮喘严重程度呈负相关（P＜0.05）,嗜酸性粒细胞、总IgE与哮喘严重程度无相关性（P＞0.05）。对轻度、中度、重度急性支气管哮喘发作的诊断效能显示：血清miR-1165-3p的曲线下面积（AUC）（0.95CI）分别为3.085（0.326～29.221）、0.712（0.611～0.829）、0.755（0.602～0.948）。血清miR-145的AUC（0.95CI）分别为0.833（0.708～0.979）、0.754（0.590～0.964）、0.816（0.671～0.993）。结论：血清miR-1165-3p、miR-145表达水平具有较高的诊断效能,支气管哮喘越严重,诊断的特异性越高,可作为支气管哮喘严重程度的无创诊断指标。     

慢性牙周炎患者血清CGRP、PGE2、CCL20与牙周临床指标和Th17/Treg失衡的相关性分析

摘要 目的：探讨慢性牙周炎（CP）患者血清降钙素基因相关肽（CGRP）、前列腺素E₂（PGE₂）、C-C基序趋化因子配体20（CCL20）与牙周临床指标和辅助性T细胞17/调节性T细胞（Th17/Treg）失衡的相关性。方法：选取2018年3月～2022年1月本院收治的92例CP患者,根据病情严重程度分为轻度组28例、中度组33例、重度组31例。检查患者牙周临床指标,采用酶联免疫吸附法检测血清CGRP、PGE₂、CCL20水平,流式细胞术检测外周血Th17、Treg百分比并计算Th17/Treg。比较3组血清CGRP、PGE₂、CCL20水平和牙周临床指标及外周血Th17、Treg、Th17/Treg,采用Pearson相关系数分析CP患者血清CGRP、PGE₂、CCL20水平与牙周临床指标和Th17/Treg的相关性。结果：轻度组、中度组、重度组血清CGRP水平和外周血Treg比例依次降低,血清PGE₂、CCL20水平和探诊深度（PD）、出血指数（BI）、附着丧失（AL）、菌斑指数（PLI）及外周血Th17比例、Th17/Treg依次升高（P＜0.05）。Pearson相关系数显示,CP患者血清CGRP水平与PD、BI、AL、PLI、Th17/Treg呈负相关,PGE₂、CCL20水平与PD、BI、AL、PLI、Th17/Treg呈正相关（P＜0.05）。结论：血清CGRP、PGE₂、CCL20水平与CP患者牙周临床指标和Th17/Treg失衡密切相关,可能是CP发生发展的关键因子。     

Heterogeneity of hemoglobin A1d: assessment and partial characterization of two new minor hemoglobins, A1d3a and A1d3b, increased in uremic and diabetic patients, respectively

We have separated and quantified two new minor hemoglobins named HbA_1d3a and HbA_1d3b. The level of HbA_1d3a was significantly higher in uremic than in non-uremic patients (3.00 ± 0.50% vs. 1.28 ± 0.26% of total hemoglobin). It correlated well with carbamylated hemoglobin (r=0.80, n=81, p<0.002) and with plasma urea concentration (r=0.78, n=81, p<0.002). These data and the electrospray ionization mass spectrometric analysis provide strong evidence that HbA_1d3a is an α-chain modified by carbamylation. The HbA_1d3b level in the diabetic patients was found to be 1.6-fold that in non-diabetic subjects (3.00 ± 0.49 vs. 1.90 ± 0.33). This was attributed to HbA_1d3 modified by glycation. Indeed HbA_1d3b correlated significantly with HbA_1c (r=0.71, p<0.002) and with serum glucose level (r=0.62, p<0.002). These two new minor hemoglobins may serve as complements for the objective assessment of averagd long-term uremia and glycemia in uremic and diabetic patients.     

Sn-S and Ru-Ru bonds cleavage reactions between [Ph3SnS(CH2)3SSnPh3] and Ru3(CO)12: X-ray crystal structures of [Ph3SnS(CH2)3SSnPh3] and trans-[Ru(CO)4(SnPh3)2]

The organotin complex [Ph₃SnS(CH₂)₃SSnPh₃] (1) was synthesized by PdCl₂ catalyzed reaction between Ph₃SnCl and disodium-1,3-propanedithiolate which in turn was prepared from 1,2-propanedithiol and sodium in refluxing THF. Reaction of 1 with Ru₃(CO)₁₂ in refluxing THF affords the mononuclear complex trans-[Ru(CO)₄(SnPh₃)₂] (2) and the dinuclear complex [Ru₂(CO)₆(μ-κ²-SCH₂CH₂CH₂S)] (3) in 20 and 11% yields, respectively, formed by cleavage of Sn-S bond of the ligand and Ru-Ru bonds of the cluster. Treatment of pymSSnPPh₃ (pymS = pyrimidine-2-thiolate) with Ru₃(CO)₁₂ at 55-60 °C also gives 2 in 38% yield. Both 1 and 2 have been characterized by a combination of spectroscopic data and single crystal X-ray diffraction analysis.     

Integrin αvβ3, metalloproteinases, and sphingomyelinase-2 mediate urokinase mitogenic effect

Plasminogen activators are implicated in the pathogenesis of several diseases such as inflammatory diseases and cancer. Beside their serine-protease activity, these agents trigger signaling pathways involved in cell migration, adhesion and proliferation. We previously reported a role for the sphingolipid pathway in the mitogenic effect of plasminogen activators, but the signaling mechanisms involved in neutral sphingomyelinase-2 (NSMase-2) activation (the first step of the sphingolipid pathway) are poorly known. This study was carried out to investigate how urokinase plasminogen activator (uPA) activates NSMase-2. We report that uPA, as well as its catalytically inactive N-amino fragment ATF, triggers the sequential activation of MMP-2, NSMase-2 and ERK1/2 in ECV304 cells that are required for uPA-induced ECV304 proliferation, as assessed by the inhibitory effect of Marimastat (a MMP inhibitor), MMP-2-specific siRNA, MMP-2 defect, and NSMase-specific siRNA. Moreover, upon uPA stimulation, uPAR, MT1-MMP, MMP-2 and NSMase-2 interacted with integrin α_vβ₃, evidenced by co-immunoprecipitation and immunocytochemistry experiments. Moreover, the α_vβ₃ blocking antibody inhibited the uPA-triggered MMPs/uPAR/integrin α_vβ₃ interaction, NSMase-2 activation, Ki67 expression and DNA synthesis in ECV304. In conclusion, uPA triggers interaction between integrin α_vβ₃, uPAR and MMPs that leads to NSMase-2 and ERK1/2 activation and cell proliferation. These findings highlight a new signaling mechanism for uPA, and suggest that, upon uPA stimulation, uPAR, MMPs, integrin α_vβ₃ and NSMase-2 form a signaling complex that take part in mitogenic signaling in ECV304 cells.     

Effects of irradiance and temperature on photosynthesis in C3, C4 and C3/C4 Panicum species

Species in the Laxa and Grandia groups of the genus Panicum are adapted to low, wet areas of tropical and subtropical America. Panicum milioides is a species with C₃ photosynthesis and low apparent photorespiration and has been classified as a C₃/C₄ intermediate. Other species in the Laxa group are C₃ with normal photorespiration. Panicum prionitis is a C₄ species in the Grandia group. Since P. milioides has some leaf characteristics intermediate to C₃ and C₄ species, its photosynthetic response to irradiance and temperature was compared to the closely related C₃ species, P. laxum and P. boliviense and to P. prionitis. The response of apparent photosynthesis to irradiance and temperature was similar to that of P. laxum and P. boliviense, with saturation at a photosynthetic photo flux density of about 1 mmol m^-2 s^-1 at 30°C and temperature optimum near 30°C. In contrast, P. prionitis showed no light saturation up to 2 mmol m^-2 s^-1 and an optimum temperature near 40°C. P. milioides exhibited low CO₂ loss into CO₂-free air in the light and this loss was nearly insensitive to temperature. Loss of CO₂ in the light in the C₃ species, P. laxum and P. boliviense, was several-fold higher than in P. milioides and increased 2- to 5-fold with increases in temperature from 10 to 40°C. The level of dark respiration and its response to temperature were similar in all four Panicum species examined. It is concluded that the low apparent photorespiration in P. milioides does not influence its response of apparent photosynthesis to irradiance and temperature in comparison to closely related C₃ Panicum species.Abbreviations AP apparent photosynthesis - I CO₂ compensation point - gl leaf conductance; gm, mesophyll conductance - PPFD photosynthetic photon flux density - PR apparent photorespiration rate - RuBPC sibulose bisphosphate carboxylase     

Synthesis of two carboxylic haptens for raising antibodies to 25-hydroxyvitamin D3 and 1α,25-dihydroxyvitamin D3

Hapten derivatives of 25-hydroxyvitamin D₃ and 1α,25-dihydroxyvitamin D₃ were synthesized using the Wittig–Horner approach. Both haptens bearing a carboxylic group at the side chain that can be linked to a protein for raising antibodies of potential utility for the determination of 25-hydroxyvitamin D₃, 1α,25-dihydroxyvitamin D₃ and 1α-hydroxylated vitamin D₃ analogues.     

Simplified method for the determination of 25-hydroxy and 1α,25-dihydroxy metabolites of vitamins D2 and D3 in human plasma application to nutritional studies

A simplified method for the determination of 25-hydroxy and 1α,25-dihydroxy metabolites of vitamins D₂ and D₃ in human plasma was developed. Plasma samples were deproteinizated and applied to a Bond Elut C₁₈ OH cartridge to separate 25-hydroxyvitamin D (25-OH-D) and 1α-25-dihydroxyvitamin D [1,25(OH)₂D] fractions. The 25-OH-D fraction was purified by a Bond Elut C₁₈ cartridge and 25-OH-D₂ and 25-OH-D₃ were assayed by HPLC using a Zorbax SIL column. The 1,25(OH)₂D fraction obtained above was subsequently applied to HPLC using a Zorbax SIL column to separate 1,25(OH)₂D₂ and 1,25(OH)₂D₃ fractions which were determined by a radioreceptor assay (RRA) using calf thymus receptor. The method was applied to nutritional studies.     

Synthesis of 2α-substituted-14-epi-previtamin D3 and its genomic activity

We synthesized and isolated 2α-substituted analogs of 14-epi-previtamin D₃ after thermal isomerization at 80 °C for the first time. The VDR binding affinity and transactivation activity of osteocalcin promoter in HOS cells were evaluated, and the 2α-methyl-substituted analog was found to have greater genomic activity than 14-epi-previtamin D₃.     

Characteristics of mass spectrometric analyses coupled to gas chromatography and liquid chromatography for 22-oxacalcitriol, a vitamin D3 analog, and related compounds

The characteristics of the mass spectra of vitamin D₃ related compounds were investigated by GC–MS and LC–MS using 22-oxacalcitriol (OCT), an analog of 1,25-dihydroxyvitamin D₃, and related compounds. Fragmentation during GC–MS (electron impact ionization) of TMS-derivatives of OCT and the postulated metabolites gave useful structural information concerning the vitamin D₃-skeleton and its side-chain, especially with respect to the oxidation positions of metabolites. In contrast, few fragment ions were observed in LC–MS (atmospheric pressure chemical ionization), showing that LC–MS gave poor structural information, except for molecular mass. However, when comparing the signal-to-noise ratio (S/N) observed during GC–MS and LC–MS analysis for OCT in plasma extracts, the S/N in LC–MS was over ten-times greater than in GC–MS, possibly due to the low recovery on derivatization and thermal-isomerization in GC–MS. Furthermore, both the GC–MS and the LC–MS allowed the analysis of many postulated metabolites in a single injection without any prior isolation of target metabolites from biological fluids by LC. These results suggest that GC–MS and LC–MS analysis for vitamin D₃ related compounds such as OCT each have unique and distinct advantages. Therefore, the complementary use of both techniques enables the rapid and detailed characterization of vitamin D₃ related compounds.