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。     

支气管哮喘患儿血清半乳糖凝集素3、类胰蛋白酶、25-羟维生素D3与肺功能和生活质量的相关性分析

摘要 目的：探讨支气管哮喘（简称"哮喘"）患儿血清半乳糖凝集素3（Gal-3）、类胰蛋白酶、25-羟维生素D₃ [25（OH）D₃]与肺功能和生活质量的相关性。方法：选取我院2018年2月～2021年3月收治的136例哮喘患儿为研究组，行肺功能检测，将其按照病情严重程度分作轻度组63例、中度组41例及重度组32例。另取同期健康体检儿童40例作为对照组。检测并比较各组血清Gal-3、类胰蛋白酶、25（OH）D₃水平。对比各组哮喘患儿肺功能指标，以中文版儿科哮喘生命质量调查问卷(PAQLQ)对哮喘患儿进行生活质量评估。并以Pearson相关性分析哮喘患儿血清Gal-3、类胰蛋白酶、25（OH）D₃水平与肺功能和生活质量的关系。结果：研究组的血清Gal-3、类胰蛋白酶水平均高于对照组，且轻度组、中度组、重度组逐渐升高；血清25（OH）D₃水平均低于对照组，轻度组、中度组、重度组逐渐下降（P＜0.05）。中度组及重度组第1秒用力呼气容积占预计值百分比（FEV₁%pred）、呼吸峰流速占预计值百分比（PEF%pred）、第1秒用力呼气容积/用力肺活量（FEV₁/FVC）以及PAQLQ各项评分均低于轻度组，且重度组低于中度组，差异均有统计学意义（P＜0.05）。经Pearson相关性分析可得：哮喘患儿血清Gal-3、类胰蛋白酶水平与各项肺功能指标以及PAQLQ各项评分均呈负相关，而血清25（OH）D₃水平与与各项肺功能指标以及PAQLQ各项评分均呈正相关（均P＜0.05）。结论：血清Gal-3、类胰蛋白酶、25（OH）D₃水平与哮喘患儿的肺功能以及生活质量密切相关，可能成为评估其病情及生活质量的可靠指标。     

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。     

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。     

术前修正衰弱指数联合血清PGE2、IL-17A预测老年髋关节置换术患者术后谵妄的临床研究

摘要 目的：探讨术前修正衰弱指数（mFI）联合血清前列腺素E₂（PGE₂）、白细胞介素-17A（IL-17A）预测老年髋关节置换术患者术后谵妄（POD）的临床研究。方法：选取2020年1月～2022年7月四川大学华西医院收治的276例老年髋关节置换术患者，根据是否发生POD分为POD组和非POD组。计算术前mFI，采用酶联免疫吸附法检测血清PGE₂、IL-17A水平。分析老年髋关节置换术患者POD的影响因素，采用受试者工作特征（ROC）曲线分析mFI和血清PGE₂、IL-17A水平对老年髋关节置换术患者POD的预测价值。结果：276例老年髋关节置换术患者POD发生率为17.03%（47/276）。与非POD组比较，POD组mFI和血清PGE₂、IL-17A水平升高（P＜0.05）。多因素Logistic回归分析显示，年龄、糖尿病、脑卒中、mFI、PGE₂、IL-17A为老年髋关节置换术患者POD的独立危险因素（P＜0.05）。ROC曲线分析显示，mFI联合血清PGE₂、IL-17A预测老年髋关节置换术患者POD的曲线下面积（AUC）大于mFI、PGE₂、IL-17A单独预测（P＜0.05）。结论：mFI和血清PGE₂、IL-17A水平升高与老年髋关节置换术患者POD独立相关，mFI联合血清PGE₂、IL-17A预测老年髋关节置换术患者POD的价值较高，可能成为老年髋关节置换术患者POD的辅助预测指标。     

消癥止痛汤联合耳穴压豆对气滞血瘀型子宫内膜异位症痛经患者炎症因子和PGE2、PGF2α的影响

摘要 目的：观察消癥止痛汤联合耳穴压豆对气滞血瘀型子宫内膜异位症（EMT）痛经患者炎症因子和前列腺素E₂（PGE₂）和前列腺素F2α（PGF2α）的影响。方法：选取2021年3月至2022年12月期间在广州中医药大学附属中山中医院收治的98例EMT痛经患者作为研究对象。采用随机数字表法将其分为对照组（常规西医治疗，n=49）和研究组（对照组的基础上消癥止痛汤联合耳穴压豆，n=49）。对比两组疗效、疼痛情况评分[疼痛视觉模拟评分（VAS）、COX痛经症状积分、]、中医证候评分、炎症因子[白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）]和PGE₂和PGF2α水平。结果：研究组的临床总有效率高于对照组（P<0.05）。治疗3个疗程后，研究组VAS、COX痛经症状积分、中医证候总评分低于对照组（P<0.05）。治疗3个疗程后，研究组IL-6、TNF-α、IL-8低于对照组（P<0.05）。治疗3个疗程后，研究组PGE₂高于对照组，PGF2α低于对照组（P<0.05）。结论：消癥止痛汤联合耳穴压豆治疗气滞血瘀型EMT痛经患者，可有效改善痛经症状，降低中医证候评分，调节血清炎症因子和PGE₂、PGF2α水平。     

苍龟探穴电针疗法联合肩关节功能锻炼对肩周炎患者肩关节功能、炎症因子和血清5- HT、PGE2的影响

摘要 目的：观察苍龟探穴电针疗法联合肩关节功能锻炼对肩周炎患者肩关节功能、炎症因子和血清5-羟色胺（5-HT）、前列腺素E₂（PGE₂）的影响。方法：选择2019年7月~2021年7月期间我院收治的肩周炎患者80例,按照入院的奇偶顺序分为对照组和研究组,各为40例,对照组接受肩关节功能锻炼,研究组接受苍龟探穴电针疗法联合肩关节功能锻炼,对比两组疗效、视觉疼痛模拟评分量表（VAS）评分、血清5- HT、PGE₂水平、肩关节活动度（前屈、后伸、外展、内收、内旋、外旋）和肩关节周围肌力相关指标、血清炎症因子水平。结果：研究组的临床总有效率高于对照组（P<0.05）。治疗后,两组VAS评分均下降,且研究组较对照组低（P<0.05）。治疗后,两组主动和被动状态下患肢前屈、后伸、内收、外展、内旋、外旋活动度均改善,且研究组优于对照组（P<0.05）。治疗后,两组前屈峰力矩（PT）、前屈平均功率（AP）、外展PT、外展AP均升高,且研究组高于对照组（P<0.05）。治疗后,两组血清PGE₂、5-HT水平均下降,且研究组低于对照组（P<0.05）。治疗后,两组血清白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、C反应蛋白（CRP）水平均下降,且研究组较对照组低（P<0.05）。结论：苍龟探穴电针疗法联合肩关节功能锻炼可促进肩周炎患者肩关节功能改善,减轻疼痛症状,作用机制可能与降低血清5- HT、PGE₂及炎症因子水平有关。     

血清APN、PGRN联合检测对脓毒症患儿疾病的预测价值及与其预后的相关性分析

摘要 目的：研究脂联素（APN）和颗粒体上皮蛋白前体（PGRN）在脓毒症患儿血清中的变化,并探讨其临床意义。方法：纳入2019年1月到2021年6月在我院接受治疗的脓毒症患儿90例,并纳入同期在我院进行体检的健康儿童30例作为对照组。根据脓毒症患儿预后将其分为存活组（n=67）和死亡组（n=23）。采集所有研究对象外周血,通过酶联免疫吸附剂测定测定血清APN、PGRN、肿瘤坏死因子α（TNF-α）、白介素1β（IL-1β）、C-反应蛋白（CRP）和降钙素原（PCT）。分析血清APN、PGRN与脓毒症患儿血清炎症因子、疾病严重程度的相关性。结果：脓毒症患儿血清APN、PGRN均显著低于对照组健康儿童（P<0.05）。在脓毒症患儿中,血清APN、PGRN与血清TNF-α、IL-1β、CRP以及PCR含量均呈负相关（P<0.05）。存活组脓毒症患儿血清APN、PGRN以及小儿危重病例评分（PCIS）均显著高于死亡组脓毒症患儿（P<0.05）,并且脓毒症患儿血清APN、PGRN与PCIS评分呈正相关。多因素Logstic回归分析结果显示：小儿危重病例评分表（PCIS）评分（OR=2.691,95%CI=1.052-4.352）、血清APN（OR=3.026,95%CI=1.628-4.692）和PGRN（OR=2.824,95%CI=1.328-6.824）水平均是影响脓毒症患儿预后的独立危险因素（P<0.05）。结论：脓毒症患儿血清APN、PGRN均显著降低,其含量与脓毒症疾病严重程度和外周血炎症反应具有相关性。     

血清miR-34b-5p、miR-155表达与早产儿急性呼吸窘迫综合征炎症因子和预后的关系分析

摘要 目的：探讨血清微小核糖核酸（miR）-34b-5p、miR-155表达与早产儿急性呼吸窘迫综合征（ARDS）炎症因子和预后的关系。方法：选择2019年2月至2021年3月我院收治的92例ARDS早产儿,根据ARDS病情严重程度将其分为轻度组（31例）、中度组（43例）和重度组（18例）,追踪患儿临床结局,根据院内死亡情况将其分为存活组（51例）和死亡组（41例）。检测所有患儿的血清miR-34b-5p、miR-155表达水平以及白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）水平,比较各组间上述指标差异,分析ARDS早产儿血清miR-34b-5p、miR-155表达与炎症因子的相关性以及miR-34b-5p、miR-155预测ARDS早产儿预后的价值。结果：重度组miR-155表达水平及IL-1β、TNF-α、IL-6水平均高于中度组和轻度组,且中度组高于轻度组（P＜0.05）,重度组miR-34b-5p表达水平低于中度组和轻度组,且中度组低于轻度组（P＜0.05）。死亡组miR-155表达水平及IL-1β、TNF-α、IL-6水平高于存活组（P＜0.05）,死亡组miR-34b-5p表达水平低于存活组（P＜0.05）。ARDS早产儿miR-155表达水平与IL-1β、TNF-α、IL-6水平均呈正相关,而miR-34b-5p表达水平与IL-1β、TNF-α、IL-6水平均呈负相关（P＜0.05）。联合miR-34b-5p、miR-155预测ARDS早产儿死亡的曲线下面积（AUC）为0.853,高于两指标单独预测的0.688、0.649。结论：ARDS早产儿血清miR-34b-5p、miR-155表达水平与患儿血清炎症因子水平以及预后有关,可作为ARDS早产儿病情评估以及预后预测的潜在指标。     

AIMS65评分联合血清GAS、PGE2、BAR对急性非静脉曲张性上消化道出血患者预后的评估价值

摘要 目的：探讨AIMS65评分联合血清胃泌素（GAS）、前列腺素E₂（PGE₂）、血尿素氮/白蛋白比值（BAR）对急性非静脉曲张性上消化道出血（ANVUGIB）患者内镜下止血治疗后预后的评估价值。方法：选取2020年5月～2022年5月北京市和平里医院消化内科收治的108例的ANVUGIB患者,根据患者住院28 d内的预后分为预后不良组和预后良好组。收集患者临床资料,检测血清GAS、PGE₂水平和计算AIMS65评分、BAR。采用单因素和多因素Logistic回归分析ANVUGIB患者内镜下止血治疗后预后不良的影响因素,受试者工作特征（ROC）曲线分析AIMS65评分、GAS、PGE₂和BAR对ANVUGIB患者内镜下止血治疗后预后不良的评估价值。结果：108例ANVUGIB患者预后不良发生率为37.96％（41/108）。单因素分析显示,预后不良组年龄大于预后良好组,心率、休克指数、AIMS65评分、GAS、BAR高于预后良好组,PGE₂水平低于预后良好组（P均＜0.05）。多因素Logistic回归分析显示,年龄增加和休克指数、AIMS65评分、GAS、BAR升高为ANVUGIB患者内镜下止血治疗后预后不良的独立危险因素,PGE₂升高为其独立保护因素（P均＜0.05）。ROC曲线分析显示,AIMS65评分联合GAS、PGE₂和BAR评估ANVUGIB患者内镜下止血治疗后预后不良的曲线下面积大于AIMS65评分、GAS、PGE₂和BAR单独评估。结论：预后不良的ANVUGIB患者AIMS65评分、GAS、BAR均高于预后良好的患者,AIMS65评分联合GAS、PGE₂和BAR评估ANVUGIB患者内镜下止血治疗后预后的价值较高。     

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.     

Transesterification of sunflower oil to biodiesel on ZrO2 supported La2O3 catalyst

ZrO₂ supported La₂O₃ catalyst prepared by impregnation method was examined in the transesterification reaction of sunflower oil with methanol to produce biodiesel. It was found that the catalyst with 21 wt% loaded La₂O₃ and calcined at 600 °C showed the optimum activity. The basic property of the catalyst was studied by CO₂-TPD, and the results showed that the fatty acid methyl ester (FAME) yield was related to their basicity. The catalyst was also characterized by TG–DTA, XRD, FTIR, SEM and TEM, and the mechanism for the formation of basic sites was discussed. It was also found that the crystallite size of support ZrO₂ decreased by loading of La₂O₃, and the model of the solid-state reaction on the surface of La₂O₃/ZrO₂ catalyst was proposed. Besides, the influence of various reaction variables on the conversion was investigated.     

应用15N示踪研究欧美杨对PM2.5无机成分NH4+和NO3-的吸收与分配

通过气溶胶发生系统模拟PM2.5颗粒的发生,运用15N示踪技术研究了欧美杨107(Populus euramericana Neva.)对PM2.5中水溶性无机成分NH+4和NO-3的吸收与分配规律。结果表明,欧美杨能够有效吸收PM2.5中的NH+4和NO-3。轻度和重度污染下,欧美杨叶片对NH+4和NO-3的吸收速率均于处理后第1天达到峰值,之后,轻度污染下对NH+4和NO-3的吸收速率迅速降低以后趋于稳定,而重度污染下对NH+4和NO-3的吸收速率缓慢下降至趋于稳定。轻度污染下的欧美杨叶片的15N含量在处理后第1天达到峰值,15N(NH+4)的含量为0.11 mg/g,干重,15N(NO-3)的为0.14 mg/g,干重,之后15N含量迅速下降至趋于稳定。重度污染下的叶片15N含量在处理第1天迅速增长,之后缓慢增长至处理后第7天达到最高值,15N(NH+4)的含量为0.11 mg/g,干重,15N(NO-3)的为0.13 mg/g,干重。处理7 d后,欧美杨不同组织器官吸收或通过再分配获取的15N含量存在差异。轻度污染下,细根对NH+4和NO-3的吸收量最高,树皮、叶柄、叶片次之,髓最低。而重度污染下,叶片对NH+4和NO-3的吸收量最高,细根、叶柄、树皮次之,髓最低。欧美杨各组织器官中NH+4和NO-3的含量均表现为重度污染大于轻度污染,且两种污染程度下的欧美杨各组织器官对NO-3的吸收均大于对NH+4的吸收。重度污染下,欧美杨茎木质部对15N(NH+4和NO-3)的吸收征调能力(Ndff,Nitrogen derived from fertilizer)最大,其次为髓,叶片最小;欧美杨各组织器官中的15N分配率表现为叶片细根叶柄树皮粗根茎木质部髓。研究结果对进一步揭示植物吸收PM2.5的机制及有效利用植物降低颗粒物污染、净化环境提供了重要的科学理论依据。     

Beta-1 integrins mediate substrate dependent effects of 1α,25(OH)2D3 on osteoblasts

Surface micron-scale and submicron scale features increase osteoblast differentiation and enhance responses of osteoblasts to 1,25-dihydroxyvitamin D₃ [1α,25(OH)₂D₃]. β₁ integrin expression is increased in osteoblasts grown on Ti substrates with rough microarchitecture, and it is regulated by 1α,25(OH)₂D₃ in a surface-dependent manner. To determine if β₁ has a role in mediating osteoblast response, we silenced β₁ expression in MG63 human osteoblast-like cells using small interfering RNA (siRNA). In addition, MG63 cells were treated with two different monoclonal antibodies to human β₁ to block ligand binding. β₁-silenced MG63 cells grown on a tissue culture plastic had reduced alkaline phosphatase activity and levels of osteocalcin, transforming growth factor β₁, prostaglandin E₂, and osteoprotegerin in comparison with control cells. Moreover, β₁-silencing inhibited the effects of surface roughness on these parameters and partially inhibited effects of 1α,25(OH)₂D₃. Anti β₁ antibodies decreased alkaline phosphatase but increase osteocalcin; effects of 1α,25(OH)₂D₃ on cell number and alkaline phosphatase were reduced and effects on osteocalcin were increased. These findings indicate that β₁ plays a major and complex role in osteoblastic differentiation modulated by either surface microarchitecture or 1α,25(OH)₂D₃. The results also show that β₁ mediates, in part, the synergistic effects of surface roughness and 1α,25(OH)₂D₃.     

Interactions between TGF-β1 and TGF-β3 and their role in medial edge epithelium cell death and palatal fusion in vitro

In recent decades, studies have shown that both TGF-β₁ and TGF-β₃ play an important role in the induction of medial edge epithelium (MEE) cell death and palatal fusion. Many of these experiments involved the addition or blockage of one of these growth factors in wild-type (WT) mouse palate cultures, where both TGF-β₁ and TGF-β₃ are present. Few studies have addressed the existence of interactions between TGF-β₁ and TGF-β₃, which could modify their individual roles in MEE cell death during palatal fusion. We carried out several experiments to test this possibility, and to investigate how this could influence TGF-β₁ and TGF-β₃ actions on MEE cell death and palatal shelf fusion. We double-immunolabelled developing mouse palates with anti-TGF-β₁ or anti-TGF-β₃ antibodies and TUNEL, added rhTGF-β₁ or rhTGF-β₃ or blocked the TGF-β₁ and TGF-β₃ action at different concentrations to WT or Tgf-β₃ null mutant palate cultures, performed in situ hybridizations with Tgf-β₁ or Tgf-β₃ riboprobes, and measured the presence of TUNEL-positive midline epithelial seam (MES) cells and MES disappearance (palatal shelf fusion) in the different in vitro conditions. By combining all these experiments, we demonstrate great interaction between TGF-β₁ and TGF-β₃ in the developing palate and confirm that TGF-β₃ has a more active role in MES cell death than TGF-β₁, although both are major inductors of MES disappearance. Finally, the co-localization of TGF-β₁, but not TGF-β₃, with TUNEL in the MES allows us to suggest a possible role for TGF-β₁ in MES apoptotic clearance.     

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.     

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.     

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 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.