感染旋毛虫昆明小鼠心肌中的脂肪酸组分及其相对含量的变化

为探讨旋毛虫致心肌炎的发生机制以及宿主抗旋毛虫感染的机制,分别取感染旋毛虫5、7、35 d后的小鼠心肌和健康小鼠心肌组织,利用AIB法提取心肌中总脂类,并经14%三氟化硼甲醇溶液进行甲酯化,做GC-MS分析.结果显示,感染旋毛虫的小鼠与健康小鼠心肌中主要脂肪酸的含量存在明显差异(P＜0.05).感染后的心肌中多不饱和脂肪酸及总n-6脂肪酸明显减少,主要是亚油酸的含量明显减少,由正常的17.23%±2.10%下降至9.21%±1.25%(P＜0.05).饱和脂肪酸则高于健康心肌的36.93%±2.53%,主要是因棕榈酸和硬脂酸的含量增加所致.受感染的心肌中C18:2和C20:4随着感染时间的延长先减少后恢复正常,而C22:5和C22:6的含量则先增加后降至正常水平.     

Automated extraction of DNA from reference samples from various types of biological materials on the Qiagen BioRobot EZ1 Workstation

We have validated and implemented a protocol for DNA extraction from various types of biological materials using a Qiagen BioRobot EZ1 Workstation. The sample materials included whole blood, blood from deceased, buccal cells on Omni swabs and FTA Cards, blood on FTA Cards and cotton swabs, and muscle biopsies. The DNA extraction was validated according to EN/ISO 17025 for the STR kits AmpF?STR^® Identifiler^® and AmpF?STR^® Yfiler^® (Applied Biosystems). Of 298 samples extracted, 11 (4%) did not yield acceptable results. In conclusion, we have demonstrated that extraction of DNA from various types of biological material can be performed quickly and without the use of hazardous chemicals, and that the DNA may be successfully STR typed according to the requirements of forensic genetic investigations accredited according to EN/ISO 17025.     

Differentiation of Morphologically Similar Human Head Hairs from Two Demographically Similar Individuals Using Amino Acid Ratios,

Human hair is frequently encountered as forensic evidence and can contribute valuable information to investigators. Conventional forensic hair analyses include microscopic hair comparison (MHC) and DNA analysis. However, MHC is not supported by statistics and DNA analysis cannot always be performed. Recent studies have demonstrated that evaluation of differences in the hair proteins may offer an alternate method to these analyses. In this study, an evaluation of the amino acids present in hair was investigated as an approach to differentiate morphologically indistinguishable hair samples from two demographically similar individuals. Proteins in the hair were digested using hydrochloric acid, and the resulting amino acids were derivatized with N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) for analysis using gas chromatography-mass spectrometry (GC-MS). Eight derivatized amino acids were detected and quantified relative to an internal standard, L-norvaline, and used to construct twenty-eight amino acid ratios. Hair samples were collected from four areas of the head on various days over the course of one month, and no significant differences in amino acid ratios (p-value > 0.05) were observed among the areas of the head, and the ratios were consistent over the time period of this study. Additionally, fifteen of these amino acid ratios were found to be significantly different between the two individuals when compared using a two-sample t-test (p-value ≤ 0.05). These data indicate that amino acid analysis was able to differentiate two morphologically similar hair samples from different individuals and demonstrates the applicability of this method to distinguish similar hair samples when DNA analysis cannot be performed.     

中药熏蒸联合针刺和透明质酸钠关节腔注射治疗骨关节炎30例

目的 观察中西医结合疗法治疗骨关节炎的疗效。方法 将60例骨关节炎患者随机分为对照组（透明质酸钠关节腔注射）和试验组（透明质酸钠关节腔注射联合中药熏蒸和针刺治疗）,每组30例。治疗5周后,采用视觉模拟评分法（visual analogue scale, VAS）测定关节疼痛程度,采用美国纽约特种外科医院膝关节评分（Hospital for Special Surgery Knee Score,HSS）评价关节功能。结果 试验组临床疗效优于对照组,但差异无统计学意义（P>0.05）;治疗后两组VAS评分均显著下降（P<0.05）,HSS评分均显著升高（P<0.05）,试验组VAS评分下降值和HSS评分升高值显著大于对照组（P＜0.05）。结论 在透明质酸钠关节腔注射治疗基础上,联合中药熏蒸和针刺治疗有利于改善关节功能,减轻疼痛。     

反向高效液相色谱法测定覆盆子中3种三萜酸的含量

目的 采用反相高效液相色谱法建立覆盆子中三萜类成分（山楂酸、科罗索酸和齐墩果酸）的测定方法,为中药覆盆子的质量评价提供依据。方法 色谱柱：WondaSil C18-WR（250 mm×4.6 mm,5 μm）,流动相为乙腈（A）-0.2%磷酸水溶液（B）（9∶1）,等度洗脱,柱温为25 ℃,流速0.6 mL/min,检测波长205 nm,进样量10 μL。结果 山楂酸、科罗索酸、齐墩果酸分别在0.156～3.900 μg、0.102～2.55 μg、2.04～51 μg范围内与峰面积呈现良好的线性关系。平均加样回收率分别为95.0%、91.7%、93.8%,RSD分别为2.59%、3.25%、2.86%。结论 该方法简单、可行,可作为覆盆子药材中单一三萜类成分含量的测定方法。     

可见分光光度法测定覆盆子中总三萜含量

目的 建立覆盆子中总三萜的含量测定方法。方法 以齐墩果酸为对照品,香草醛-冰醋酸-高氯酸为显色体系,采用可见分光光度法在480 nm处测定覆盆子中总三萜的含量。〖JP〗结果 齐墩果酸在0.079 60～0.398 0 mg范围内与吸光度呈良好的线性关系,回归方程为y＝2.117x＋0.005 380（r＝0.999 7）,平均加样回收率为101.92%(RSD＝1.92%,n＝6)。结论 该方法操作简便、快速、灵敏度高、重现性好、测定结果准确,可作为覆盆子中总三萜含量测定的方法。     

新藤黄酸对人脐静脉内皮细胞凋亡的影响

目的 探讨新藤黄酸对人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）增殖和凋亡的影响。方法 用不同浓度的新藤黄酸培养细胞24、48 h后,采用MTT法观察新藤黄酸对细胞生长的抑制作用;倒置显微镜下观察HUVECs细胞形态变化;通过DAPI染色后倒置荧光显微镜观察新藤黄酸对细胞形态学的影响;通过Annexin V-FITC/PI双染及流式细胞术检测新藤黄酸对HUVECs细胞凋亡率的影响;Western blot检测新藤黄酸对HUVECs中凋亡相关蛋白Caspase-3的表达变化。结果 MTT检测结果显示新藤黄酸能够明显抑制HUVECs的增殖,并呈时效及量效关系;荧光显微镜观察结果显示,新藤黄酸作用HUVECs后出现大量凋亡细胞;流式细胞术结果显示,与空白对照组相比,随着新藤黄酸浓度的增加,细胞凋亡率明显增加;Western blot检测结果表明Caspase-3蛋白表达水平显著增加。结论 新藤黄酸对HUVECs有明显的抑制作用,其作用机制可能是通过诱导细胞凋亡实现的。     

多指标正交试验优选苓桂术甘颗粒水提工艺

目的 优选苓桂术甘颗粒的水提工艺。方法 采用正交试验对水提工艺进行优化,以桂皮醛转移率、甘草酸转移率和干膏得率为评价指标,考察加水量、煎煮时间、煎煮次数对提取工艺的影响。结果 最佳提取工艺为加8倍量水,提取3次,每次煎煮1 h。结论 多指标加权考察水提工艺,既符合中药复方的复杂性,又能合理反映水提效果。该水提工艺设计合理,可为苓桂术甘颗粒的研制提供理论和试验基础。     

茯苓不同药用部位化学成分分析

目的 比较茯苓不同药用部位化学成分的含量,为茯苓药材产地加工的产业化研究和临床用药提供参考依据。方法 分别采用紫外分光光度法、高效液相色谱法测定茯苓不同药用部位中水不溶性多糖、茯苓酸的含量。结果 茯苓不同药用部位的水不溶性多糖含量大小依次为白茯苓、茯神、赤茯苓、茯苓皮,茯苓酸含量大小依次为赤茯苓、茯苓皮、白茯苓、茯神。结论 茯苓不同药用部位的化学成分分布存在一定的差异,茯苓酸和茯苓水不溶性多糖的含量与临床用药的选择有关联;所建立的多指标成分含量测定方法准确可靠,可用于茯苓药材质量的评价。     

桃红四物汤对产后血瘀证大鼠血清溶血磷脂酸的影响

目的 探讨桃红四物汤对产后血瘀证大鼠血清溶血磷脂酸(lysophosphatidic acid,LPA)水平的影响。方法 采用米非司酮（8.3 mg/kg）和米索前列醇（0.1 mg/kg）灌胃早孕鼠,复制血瘀证大鼠模型,将模型复制成功的大鼠随机分为模型组、益母草颗粒组（4.3 g/kg）及桃红四物汤高（18.0 g/kg）、中（9.0 g/kg）、低（4.5 g/kg）剂量组,并设立正常对照组;测定血液流变学指标以判定桃红四物汤治疗血瘀证的疗效,采用酶联免疫吸附试验测定血清LPA含量。结果 模型组全血黏度、血浆黏度、血清LPA均较正常对照组显著增高（P＜0.05,或P＜0.01）;桃红四物汤中剂量组及益母草颗粒组大鼠全血低切变率黏度和高切变率黏度显著降低（P＜0.05）,各药物组大鼠血浆黏度显著降低（P＜0.05,或P＜0.01）;桃红四物汤中剂量组及益母草颗粒组大鼠LPA水平显著降低（P＜0.05）。结论 桃红四物汤祛瘀作用显著,其治疗作用可能与降低血清LPA的水平有关。