秦艽清虚热的质疑与探析

质疑秦艽清虚热的功效。从秦艽功用沿革及历代医家对秦艽的认识,总结秦艽功效为主传尸骨蒸、除日晡潮热骨蒸、治小儿疳积骨蒸,而秦艽的除骨蒸作用不能等同于清虚热,秦艽并不具备治疗阴虚发热的特质,其在治疗阴虚发热方剂中的作用与其性用平和、辛散苦泄通利有关,而它本身并无直接的清退阴虚发热的功用。     

胃泌素释放肽前体双抗体夹心酶联免疫吸附法的建立及其应用

目的：胃泌素释放肽前体（ pro-gastrin releasing peptide , PGRP）作为小细胞肺癌肿瘤标志物的应用价值已成为近年来的研究热点,文中建立新型双抗体夹心酶联免疫吸附法（ enzyme-linked immune sorbent assay , ELISA ）方法检测患者血清PGRP抗原浓度。方法通过人工合成PGRP抗原决定簇对本实验室自行研制的单克隆抗体进行筛选;采用改良过碘酸钠法对筛选到的单克隆抗体进行辣根过氧化物酶标记,建立PGRP双抗体夹心酶联免疫吸附检测法;并与IBL公司PGRP试剂盒对比检测结果。结果新型ELISA检测标准抗原浓度范围为33～1．7×104 pg／mL,检测小细胞肺癌患者血清PGRP浓度灵敏度100％,特异度98．4％,IBL试剂盒检测灵敏度100％,特异度92．2％。结论新型ELISA方法可用于小细胞肺癌患者血清PGRP浓度检测。     

阿霉素化学键接聚合物纳米颗粒的合成及pH敏感释放

合成了前驱体三-(2-氨乙基)-N,N',N"-三硫辛酸氨,该前驱体经三丁基膦(TBUP)还原后得到末端带有6个巯基的小分子单体(C-6SH)并共价偶联腙键修饰的阿霉素-(6-马来酰亚胺基己酰)腙(MAL-DOX)。通过巯基与丙烯酸酯的点击化学反应,以二甲基丙烯酸乙二醇酯(EGDMA)为交联剂,连续投料制备了具有内核交联、表面聚乙二醇(PEG)化与pH敏感药物释放特性的聚合物纳米颗粒。采用动态光散射粒径分析(DLS)和透射电镜(TEM)对纳米颗粒的尺寸和形貌进行表征,结果表明该纳米颗粒呈球形,平均尺寸为42 nm。体外药物释放表明:通过腙键连接阿霉素(DOX)的聚合物纳米颗粒具有良好的pH响应性释放能力,即在酸性条件下(pH=5.5)的释放速率比生理pH环境下要明显加快。     

生长激素释放激素受体激动剂MR409减轻糖尿病Db/Db小鼠血管钙化

目的研究生长激素释放激素受体激动剂MR409对基因缺陷小鼠Db/Db糖尿病小鼠(Db/Db小鼠)血管钙化(VC)的保护作用。方法以12周龄雄性野生型(WT)C57BL/6小鼠为WT组(对照组),12周龄雄性Db/Db小鼠随机分为Db/Db组(糖尿病组)和MR409组(MR409治疗组)。MR409组隔天给予MR409(15μg/次)治疗8周。第8周末,检测小鼠血清碱性磷酸酶(ALP)和血糖水平。HE染色观察小鼠主动脉形态变化。von Kossa染色、茜素红染色检测主动脉钙盐沉积。离体血管灌流系统测定内皮依赖性血管舒张功能。免疫组织化学染色检测主动脉钙化相关蛋白Runt相关转录因子2(Runx2)蛋白表达。二氢乙锭荧光染色检测主动脉活性氧(ROS)水平。结果与WT组比较,Db/Db组血清ALP活性升高,血管壁厚度增加,血管舒张功能降低,主动脉ROS水平增加(P0.05);与Db/Db组比较,MR409组ALP活性降低,主动脉肥厚减轻,血管舒张功能改善,主动脉ROS水平降低(P0.05)。与WT组比较,Db/Db组有明显的VC,表现为主动脉钙盐沉积增加,Runx2蛋白表达升高(P0.05);与Db/Db组比较,MR409组主动脉钙盐沉积减轻,Runx2蛋白表达降低(P0.05)。结论 MR409能显著抑制糖尿病小鼠VC,减轻主动脉增厚,改善血管内皮功能,其机制可能与抑制氧化应激诱导的Runx2表达升高有关。     

Oxygen toxicity in the infant rhesus monkey: effects on regulatory peptides in lung and blood

A total of ten 6-month-old male rhesus monkey (Macaca mulatta) infants, born full-term, were positive-pressure ventilated with greater than 95% oxygen or room air (controls). A protocol was used which closely simulated pediatric intensive care. To test if regulatory peptides were affected by the oxygen treatment, and to search for an early marker of oxygen toxicity, lung tissue samples and systemic mixed venous blood were collected at 6, 12 and 24 hours after onset of treatment. The peptides, gastrin releasing peptide (GRP), calcitonin gene-related peptide (CGRP), peptide YY (PYY), vasoactive intestinal peptide (VIP) and somatostatin (SOM), were quantitated in lung tissue extracts and plasma using radioimmunoassay. Lung tissue GRP, CGRP, and PYY levels appeared to decrease gradually with time, perhaps as a result of the positive pressure ventilation procedure. GRP and CGRP levels decreased less among monkey infants ventilated with oxygen, thus they were significantly higher at 24 hours than in air ventilated controls. VIP levels were significantly lower among tests compared to controls at that time. Blood peptide levels did not change with oxygen treatment. These results suggest that tissue concentrations of certain pulmonary regulatory peptides can become altered by ventilation with greater than 95% oxygen. A blood borne peptide marker was not identified.     

蛙皮素对人胃黏膜上皮永生细胞系GES-1细胞生长作用和机制的研究

目的　观察蛙皮素对人永生细胞系 (GES 1)生长的调节和作用机制。方法　①分子水平观察GES 1细胞胃泌素释放肽 (GRP)受体mRNA的表达。②利用受体化学交联和细胞膜组织化学法观察GRP受体蛋白表达。③观察不同浓度的蛙皮素及其受体拮抗剂对GES 1细胞生长的影响。④观察蛋白激酶C (PKC)抑制剂对蛙皮素促进GES 1细胞作用的影响。⑤观察蛙皮素作用GES 1细胞后 ,细胞内三磷酸肌醇 (IP3 )和PKC活性以及观察蛙皮素对此细胞系作用后GRP受体mRNA表达量的变化。结果　①GES 1细胞表达GRP受体mRNA ,Southern杂交进一步证明表达特异性mRNA。②GRP受体蛋白相对分子质量为 75× 10 3 ,该受体位于细胞膜。③蛙皮素可促进此细胞生长 ,而蛙皮素特异性受体拮抗剂可抑制此促生长作用。④蛙皮素对该细胞的促生长作用可被PKC抑制剂所抑制。⑤蛙皮素可引起细胞内IP3 含量显著增加以及引起细胞内PKC活性自细胞质向细胞膜转移。⑥蛙皮素作用GES 1细胞后可引起GRP受体mRNA表达增加。结论　此研究证明人胃黏膜上皮细胞系GES 1表达GRP受体mRNA和蛋白。蛙皮素对GES 1细胞的促生长作用是通过特异性受体介导 ,引起细胞内IP3增加和PKC活性转移来实现的。蛙皮素作用后引起该细胞GRP受体mRNA表达量增加 ,说明蛙皮素对其自身受体有上调作用     

Detection of bioactive peptides including gonadotrophin‐releasing factors (GnRHs) in horse urine using ultra‐high performance liquid chromatography–high resolution mass spectrometry (UHPLC/HRMS)

The use of bioactive peptides as a doping agent in both human and animal sports has become increasingly popular in recent years. As such, methods to control the misuse of bioactive peptides in equine sports have received attention. This paper describes a sensitive accurate mass method for the detection of 40 bioactive peptides and two non‐peptide growth hormone secretagogues (< 2 kDa) at low pg/mL levels in horse urine using ultra‐high performance liquid chromatography‐high resolution mass spectrometry (UHPLC/HRMS). A simple mixed‐mode cation exchange solid‐phase extraction (SPE) cartridge was employed for the extraction of 42 targets and/or their in vitro metabolites from horse urine. The final extract was analyzed using UHPLC/HRMS in positive electrospray ionization (ESI) mode under both full scan and data independent acquisition (DIA, for MS²). The estimated limits of detection (LoD) for most of the targets could reach down to 10 pg/mL in horse urine. This method was validated for qualitative detection purposes. The validation data, including method specificity, method sensitivity, extraction recovery, method precision, and matrix effect were reported. A thorough in vitro study was also performed on four gonadotrophin‐releasing factors (GnRHs), namely leuprorelin, buserelin, goserelin, and nafarelin, using the S9 fraction isolated from horse liver. The identified in vitro metabolites have been incorporated into the method for controlling the misuse of GnRHs. The applicability of this method was demonstrated by the identification of leuprorelin and one of its metabolites, Leu M4, in urine obtained after intramuscular administration of leuprorelin to a thoroughbred gelding (castrated horse).     

Age‐stratified and gender‐specific reference intervals of six tumor markers panel of lung cancer: A geographic‐based multicenter study in China

BackgroundSerum biomarkers have been widely adopted in clinical practice for assisting lung cancer diagnoses, therapeutic monitoring, and prognostication. The function of a well‐performing tumor biomarker depends on a reliable reference interval (RI) with consideration of the study subjects’ age, gender, and geographical location. This study aimed to establish a RI for each of 6 lung cancer biomarkers for use in the whole country of China on Mindray platform.MethodsThe levels of serum 6 lung cancer biomarkers—namely progastrin‐releasing peptide (ProGRP), neuron‐specific enolase (NSE), squamous cell carcinoma antigen (SCC), carcinoembryonic antigen (CEA), cytokeratin‐19 fragment (CYFRA21‐1), and human epididymis protein 4 (HE4)—were measured utilizing the chemiluminescence immunoassay on the Mindray CL‐6000i platform following the laboratory standard operating procedures in apparently healthy Chinese individuals on large cohort, multicenter, and geographical consideration bases. The CLSI EP28‐A3C guideline was followed for the enrollment of study subjects.ResultsThe age‐stratified, gender‐specific RIs for ProGRP, NSE, SCC, CEA, CYFRA21‐1, and HE4 lung cancer biomarkers in the Chinese population have been established as described in the results and discussion in this work. In addition, various levels of the six lung cancer biomarkers among nine geographical locations in China have been observed.ConclusionsThe sample volume of study cohort, age, and geographical location should be considered upon establishing a reliable biomarker RI. A RI for each of six lung cancer biomarkers has been established. The results from this study would be helpful for clinical laboratories in interpreting the analytical results and for clinicians in patient management.     

全关节镜下改良Broström术治疗慢性踝关节外侧不稳的临床疗效观察

目的 探讨全关节镜下改良Broström术治疗慢性踝关节外侧不稳的临床疗效。方法 回顾性研究。纳入2016年1月—2018年6月徐州市中心医院慢性踝关节外侧不稳患者33例(33侧),其中男20例、女13例,年龄16～55(30.3±5.7)岁;右侧22例,左侧11例。病程12~48(25.6±14.2)个月。受伤原因:扭伤28例,车祸伤5例。所有患者进行全关节镜下的改良Broström手术。术后定期随访,观察患者伤口愈合情况,并发症发生情况,观察踝关节活动情况,踝关节内翻应力试验、踝关节前抽屉试验检查踝关节稳定情况及步态。术后12个月采用疼痛视觉模拟评分法(VAS)评价踝关节疼痛情况,采用美国足踝外科协会(AOFAS)踝与后足评分和Karlsson 踝关节功能评分(KAFS)评价踝关节功能。对比患者手术前与术后12个月VAS、AOFAS、KAFS评分改善情况。结果 33例患者均顺利完成手术,手术时间35～80(50.3±12.6)min。术后患者切口均一期愈合,未出现伤口感染、神经损伤、血管损伤、肌腱损伤等并发症。33例患者均获得随访,随访时间12～40(25.0±10.7)个月。末次随访时,患者均无置入物排斥反应或线结反应;踝关节疼痛完全消失,关节活动度良好;踝关节内翻应力试验、踝关节前抽屉试验均阴性,未出现踝关节外侧不稳定,患者均恢复正常步态。术后12个月依据AOFAS评分标准,踝关节功能优27例、良6例;依据KAFS评分标准,踝关节功能优28例、良5例。术后12个月VAS评分由术前(6.02±1.93)分降低到(1.33±0.16)分,AOFAS评分由术前(61.75±13.08)分提高到(92.37±4.15)分,KAFS评分由术前(64.46±10.81)分提高到(93.19±5.58)分,差异均有统计学意义(t=12.765, -13.303, -12.346, P值均＜0.05)。结论 全关节镜下改良Broström术,手术创伤小,是一种治疗慢性踝关节外侧不稳的有效方法。