藏药石榴莲花散中鞣花酸和胡椒碱含量测定方法研究

目的： 建立藏药石榴莲花散中鞣花酸和胡椒碱含量测定方法,以完善其质量标准。方法： 采用高效液相色谱法测定制剂中石榴子和诃子的鞣花酸含量,采用ODS-2 HYPERSIL C₁₈色谱柱（4.6 mm×250mm,5 μm）,以乙腈-0.2%磷酸（15：85）为流动相,流速1.0 mL·min^-1,检测波长254 nm,柱温30℃;采用高效液相色谱法测定制剂中荜茇的胡椒碱含量,采用ODS-2 HYPERSIL C₁₈色谱柱（4.6mm×250 mm,5μm）,以乙腈-水（40：60）为流动相,流速1.0 mL·min^-1,检测波长343 nm,柱温30℃。结果： 鞣花酸进样量在5.065~25.325 μg范围内呈良好线性关系（r=0.9999）,平均回收率为97.5%,RSD=1.5%（n=6）;胡椒碱进样量在10.414~52.071 μg范围内呈良好线性关系（r=1.0000）,平均回收率为99.6%,RSD=0.6%（n=6）。结论： 该方法可准确测定石榴莲花散中鞣花酸和胡椒碱的含量,有助于提高石榴莲花散的质量标准。     

不同包装和贮藏条件下酸枣仁的质量比较

目的:确定酸枣仁适合的贮藏条件、包装材料和包装方式,为保障酸枣仁质量提供参考。方法:选用同一批酸枣仁样品,分别采取塑料编织袋包装、塑料袋包装、塑料袋真空包装、铝塑复合袋包装、铝塑复合袋真空包装和牛皮淋膜纸袋包装,贮藏在阴凉库(温度≤20℃、相对湿度为45%~75%)和药品稳定性试验箱[温度为(40±2)℃、相对湿度为(75±5)%]中,以性状及水分、黄曲霉毒素、酸枣仁皂苷A、斯皮诺素含量为考察指标,开展为期6个月的阴凉长期稳定性试验和加速稳定性试验研究。结果:6个月阴凉长期稳定性试验结果显示,不同包装条件下酸枣仁样品的性状以及水分、黄曲霉毒素、酸枣仁皂苷A含量均符合2015年版《中国药典》(一部)(后文简称"药典")酸枣仁项下规定;斯皮诺素含量均不符合药典的规定(不低于0.080%),但其中铝塑复合袋真空包装样品中斯皮诺素含量(0.079%)与药典要求接近。6个月加速稳定性试验结果显示,牛皮淋膜纸袋包装、塑料编织袋包装样品发霉严重,塑料袋包装、塑料袋真空包装样品表面颜色变暗,铝塑复合袋包装样品颜色稍变暗,但铝塑复合袋真空包装样品外观基本无变化;塑料编织袋包装、牛皮淋膜纸袋包装样品中水分含量超过了药典要求最高值(9.0%);仅在贮藏2个月时,编织塑料袋包装样品被检出黄曲霉毒素B1含量为8.64μg/kg,超出药典规定;各包装样品中酸枣仁皂苷A含量虽有下降,但均满足药典要求;仅塑料袋真空包装样品中斯皮诺素含量(0.084%)满足药典要求,其次属铝塑复合袋真空包装样品中含量(0.071%)相对较高。结论:酸枣仁以铝塑复合袋真空包装后置于阴凉干燥处为宜。     

Logistic模型联合ROC曲线对利奈唑胺致老年患者血小板减少的预测分析

目的：采用Logistic模型和ROC曲线探讨老年患者接受利奈唑胺治疗引起血小板减少的相关危险因素,并预测该人群中不良反应的发生风险。方法：回顾性分析某院2018年3月至2019年9月期间接受利奈唑胺治疗老年患者的临床资料,收集患者的基本信息、实验室指标、联合用药和不良反应的情况。采用多因素Logistic回归法分析接受利奈唑胺治疗的老年患者发生血小板减少症的危险因素,通过建立Logistic模型,并联合ROC曲线对血小板减少症的发生情况进行预测分析。结果：102例患者（男性68名,女性34名）纳入研究,利奈唑胺治疗期间23例（22.5%）发生血小板减少症。多因素Logistic回归分析显示用药时间,血肌酐浓度和血小板基线值是发生血小板减少症的独立危险因素。分别用上述独立危险因素建立Logistic回归方程,经变换后得到联合预测因子的计算公式,Y联合=X_Scr+34.0X_T-1.6X_PLT,联合预测因子的ROC曲线下面积（0.821,95%CI：0.755~0.866,P<0.001）优于其他指标,具有一定的预测价值,Youden指数最大时（0.606）的切点为ROC曲线上的最佳界值（183.7）。结论：用药时间、基线血小板值、血肌酐浓度是利奈唑胺致老年患者血小板减少的独立危险因素,临床用药期间,可将上述因素带入联合预测因子的计算公式,预测发生血小板减少症的风险,以便及时调整给药方案。     

髋关节数字化断层融合摄影的低剂量研究

目的 探讨髋关节数字化断层融合(DTS)摄影中最佳剂量比的设定,以最大程度地减少患者接受的辐射剂量.方法 90例行髋关节数字化断层融合摄影的患者,以随机数字表法,分别以6、7、8倍的剂量比组进行DTS检查,使用单因素方差分析比较不同剂量比设定下所得的图像质量和辐射剂量.结果 3组的表面吸收剂量(ESD)、剂量面积乘积(DAP)、图像质量评分分别为[(3.76±1.89)mGy、(18.41±11.71)dGy ·cm²、3.03±0.24]、[(5.24±2.76)mGy、(26.99±13.34)dGy ·cm²)、3.60±0.11]、[(6.39±1.75)mGy、(36.96±22.49)dGy ·cm²、3.64±0.09],各组间差异有统计学意义(F=10.94、9.45、139.26, P<0.05),通过3组间两两比较,ESD值和DAP值均是6倍剂量比组<7倍剂量比组<8倍剂量比组.6倍剂量比组的图像质量评分低于其他两组,7倍剂量比组和8倍剂量比组的图像质量评分差异无统计学意义.结论 剂量比是决定DTS检查辐射剂量的重要参数,髋关节DTS检查的剂量比设置为7时,可实现低曝光剂量和高图像质量的平衡,以最大限度地保护患者免受不必要的辐射损害.     

腹盆腔放疗诱发肠道微生态失调与肠源性感染的实验研究

目的 探索腹盆腔放疗照射对肠道微生态的影响及其与肠源性感染的关系。方法 模拟腹盆腔放疗照射BALB/c小鼠,2.0 Gy/d,连续照射5 d/周,分别于照射3周、5周和6周后停照1周的时间点收集回肠组织及其内容物样本。用实时定量RT-PCR检测抗菌肽和促炎性因子的表达;用PCR检测细菌在小鼠体内的移位情况;用变性梯度凝胶电泳技术检测分析肠道微生态的特征。结果 腹盆腔照射诱发了肠道潘氏细胞隐窝素-1和-4表达紊乱,照射3周或照射6周后停照1周,小鼠回肠隐窝素-1和-4均呈现显著性降低(t=-7.43、-3.54、-4.72、-4.27,P<0.05);而照射5周小鼠回肠隐窝素-1和-4表达明显升高(t=6.15、5.75,P<0.05)。放疗模拟照射3和5周时小鼠肠道微生物区系多样性指数和丰富度显著降低(t=-3.49、-4.19、-3.44、-4.97,P<0.05),呈现以乳酸杆菌等益生菌减少,大肠杆菌和弗氏志贺氏菌等条件致病菌增多为特征的微生态失调。受照小鼠肠系膜淋巴结和血液中的细菌DNA阳性率明显增高。照射3和5周后回肠组织IL-1β、IL-6和TNF-α显著性高表达(t=4.85、6.16、7.71、4.60、4.86、5.97,P<0.05);照射6周后停照1周时,肠道促炎性因子的表达量有所回落,但IL-1β和TNF-α的表达量仍显著性高表达(t=3.67、5.88,P<0.05)。结论 腹盆腔放疗可诱发肠道抗菌肽表达紊乱,引起肠道微生态失调,进而导致肠源性细菌移位及感染性炎症的发生。微生态可能成为减轻放疗患者消化道不良反应的有效干预靶点。     

小儿腹泻外敷散对腹泻大鼠小肠Cajal细胞超微结构及血管活性肠肽受体1表达的影响

目的探讨小儿腹泻外敷散对腹泻大鼠小肠Cajal间质细胞(ICC)和血管活性肠肽受体1(VIP-R1)表达的影响。方法 30只Wistar大鼠随机分为对照组、模型组、药物组,每组10只。采用番泻叶水煎液灌胃(2ml/100g)的方法建立腹泻模型,药物组给予小儿腹泻外敷散贴敷。采用透射电镜观察药物对腹泻大鼠小肠ICC超微结构的影响,免疫组化法观察VIP-R1在腹泻大鼠小肠内的分布,RT-PCR和Western blotting测定VIP-R1 m RNA和蛋白表达水平。结果药物贴敷可使ICC形态和缝隙连接基本恢复正常。VIP-R1主要分布在小肠组织环形肌与纵形肌之间、小肠组织肌间神经丛处、结肠黏膜层上皮细胞及固有层细胞的周围。小儿腹泻外敷散可抑制VIP-R1分泌并下调VIP-R1 m RNA及蛋白表达。结论小儿腹泻外敷散可修复腹泻大鼠损伤的ICC细胞,同时可通过下调VIP-R1表达而治疗腹泻。     

多次精液优化对IUI成功率的影响

目的探讨多次精液优化对宫腔内人工授精(IUI)成功率的影响。方法回顾分析我院生殖医学中心41例多次精液优化人工授精周期,分析多次精液优化对妊娠率的影响。结果 41例多次精液优化人工授精周期妊娠率为29.27%,对照组为11.62%,两组临床妊娠率比较,差异有统计学意义(P<0.05)。结论多次精液优化人工授精是一种可以提高人工授精周期妊娠率的方法,具有积极的临床意义。     

2型糖尿病患者血清IGF-1水平与颈动脉粥样斑块的相关性研究

目的探讨2型糖尿病(T2DM)患者血清胰岛素样生长因子-1(IGF-1)水平与双侧颈动脉粥样硬化斑块的相关性。方法应用超声检测186例患者双侧颈动脉内膜-中膜厚度,根据超声检查结果将其分为:(1)A组(颈动脉粥样硬化斑块阳性的T2DM患者)96例;(2)B组(颈动脉粥样硬化斑块阴性的T2DM患者)90例。并采用免疫放射分析法检测两组患者的血清IGF-1水平。结果 A组患者的血清IGF-1水平与B组比较显著降低,差异有显著统计学意义(P<0.01);T2DM患者血清IGF-1水平与双侧颈动脉粥样斑块的程度呈显著负相关(r=0.649,P<0.01)。结论 IGF-1参与了T2DM大血管病变的发生、发展,IGF-1水平与颈动脉粥样斑块程度密切相关。     

Can Cytoprotective Cobalt Protoporphyrin Protect Skeletal Muscle and Muscle-derived Stem Cells From Ischemic Injury?

Background

Extremity trauma is the most common injury seen in combat hospitals as well as in civilian trauma centers. Major skeletal muscle injuries that are complicated by ischemia often result in substantial muscle loss, residual disability, or even amputation, yet few treatment options are available. A therapy that would increase skeletal muscle tolerance to hypoxic damage could reduce acute myocyte loss and enhance preservation of muscle mass in these situations.

Questions/purposes

In these experiments, we investigated (1) whether cobalt protoporphyrin (CoPP), a pharmacologic inducer of cytoprotective heme oxygenase-1 (HO-1), would upregulate HO-1 expression and activity in skeletal muscle, tested in muscle-derived stem cells (MDSCs); and (2) whether CoPP exposure would protect MDSCs from cell death during in vitro hypoxia/reoxygenation. Then, using an in vivo mouse model of hindlimb ischemia/reperfusion injury, we examined (3) whether CoPP pharmacotherapy would reduce skeletal muscle damage when delivered after injury; and (4) whether it would alter the host inflammatory response to injury.

Methods

MDSCs were exposed in vitro to a single dose of 25 μΜ CoPP and harvested over 24 to 96 hours, assessing HO-1 protein expression by Western blot densitometry and HO-1 enzyme activity by cGMP levels. To generate hypoxia/reoxygenation stress, MDSCs were treated in vitro with phosphate-buffered saline (vehicle), CoPP, or CoPP plus an HO-1 inhibitor, tin protoporphyrin (SnPP), and then subjected to 5 hours of hypoxia (< 0.5% O₂) followed by 24 hours of reoxygenation and evaluated for apoptosis. In vivo, hindlimb ischemia/reperfusion injury was produced in mice by unilateral 2-hour tourniquet application followed by 24 hours of reperfusion. In three postinjury treatment groups (n = 7 mice/group), CoPP was administered intraperitoneally during ischemia, at the onset of reperfusion, or 1 hour later. Two control groups of mice with the same injury received phosphate-buffered saline (vehicle) or the HO-1 inhibitor, SnPP. Myocyte damage in the gastrocnemius and tibialis anterior muscles was determined by uptake of intraperitoneally delivered Evans blue dye (EBD), quantified by image analysis. On serial sections, inflammation was gauged by the mean myeloperoxidase staining intensity per unit area over the entirety of each muscle.

Results

In MDSCs, a single exposure to CoPP increased HO-1 protein expression and enzyme activity, both of which were sustained for 96 hours. CoPP treatment of MDSCs reduced apoptotic cell populations by 55% after in vitro hypoxia/reoxygenation injury (from a mean of 57.3% apoptotic cells in vehicle-treated controls to 25.7% in CoPP-treated cells, mean difference 31.6%; confidence interval [CI], 28.1–35.0; p < 0.001). In the hindlimb ischemia/reperfusion model, CoPP delivered during ischemia produced a 38% reduction in myocyte damage in the gastrocnemius muscle (from 86.4% ± 7% EBD⁺ myofibers in vehicle-treated, injured controls to 53.2% EBD⁺ in CoPP-treated muscle, mean difference 33.2%; 95% CI, 18.3, 48.4; p < 0.001). A 30% reduction in injury to the gastrocnemius was seen with drug delivery at the onset of reperfusion (to 60.6% ± 13% EBD⁺ with CoPP treatment, mean difference 25.8%; CI, 12.2–39.4; p < 0.001). In the tibialis anterior, however, myocyte damage was decreased only when CoPP was given at the onset of reperfusion, resulting in a 27% reduction in injury (from 78.8% ± 8% EBD⁺ myofibers in injured controls to 58.3% ± 14% with CoPP treatment, mean difference 20.5%; CI, 6.1–35.0; p = 0.004). Delaying CoPP delivery until 1 hour after tourniquet release obviated the protective effect in both muscles. Mean MPO staining intensity per unit area, indicating the host inflammatory response, decreased by 27–34% across both the gastrocnemius and tibialis anterior muscles when CoPP was given either during ischemia or at the time of reperfusion. Delaying drug delivery until 1 hour after the start of reperfusion abrogated this antiinflammatory effect.

Conclusions

CoPP can decrease skeletal muscle damage when given early in the course of ischemia/reperfusion injury and also provide protection for regenerative stem cell populations.

Clinical Relevance

Pharmacotherapy with HO-1 inducers, delivered in the field, on hospital arrival, or during trauma surgery, may improve preservation of muscle mass and muscle-inherent stem cells after severe ischemic limb injury.

Electronic supplementary material

The online version of this article (doi:10.1007/s11999-015-4332-8) contains supplementary material, which is available to authorized users.