HPLC法同时测定健儿膏中8种成分

目的建立HPLC法同时测定健儿膏(党参、山药、甘草等)中白术内酯Ⅲ、白术内酯Ⅰ、党参炔苷、紫丁香苷、去氢土莫酸、土莫酸、去氢茯苓酸、茯苓酸的含有量。方法该药物甲醇提取液的分析采用Diamonsil Plus C18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.05%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长210、220、266 nm。结果 8种成分在各组范围内线性关系良好(r≥0.999 1),平均加样回收率96.99%~100.01%,RSD 0.83%~1.47%。结论该方法准确可靠,重复性好,可用于健儿膏的质量控制。     

可溶性细胞间黏附分子-1在急性肝内胆汁淤积兔模型中的表达

目的探讨可溶性细胞间黏附分子(sICAM-1)在急性肝内胆汁淤积兔模型中的表达及其意义。方法39只健康的新生大耳白兔随机分成4组:3个实验组(n=9)和1个空白对照组(n=12)。每个实验组分别为3个时间点处理组(24、48、72 h)。实验组采用1次灌服α-荼异硫氰酸盐(ANIT)200 mg/kg方法造成急性肝内胆汁淤积。按时分批取标本,检测其血清和胆汁中sICAM-1水平。结果实验组血清、胆汁中sICAM-1各时间点水平明显高于正常对照组(P均<0.05),其中48 h>72 h>24 h。结论sICAM-1在胆汁淤积中表达升高,与肝脏损伤程度和病程相关。     

MicroRNAs及其在泌尿系肿瘤的研究进展

microRNAs（miRNAs）是一类分布十分广泛的内源性非编码RNA,在动物、植物、病毒中广泛存在。miRNAs与肿瘤的发生发展预后有关,并在肿瘤的增殖、分化及调亡等方面有重要的作用。结合最近相关文献,本文就miRNAs与泌尿系肿瘤方面的进展作简要概述。     

Extracellular signal regulated kinases 1/2 signal pathway and responses of astrocytes after diffuse brain injury

BACKGROUND: The treatment of diffuse brain injury during an acute period is focused on relieving degrees of secondary brain injury. Generation and development of pathological changes of secondary brain injury depend on signal conduction, so down-regulating over response of astrocyte through interfering a key link of signal conduction pathway may bring a new thinking for the treatment of diffuse brain injury. OBJECTIVE: To observe the effect of over activity of extracellular signal regulated kinases 1/2 (ERK1/2) signal pathway on the response of astrocyte during an acute period of diffuse brain injury. DESIGN: Completely randomized grouping and controlled animal study. SETTINGS: Department of Neurosurgery, the Third Affiliated Hospital, Nanchang University; Department of Neurosurgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: A total of 158 healthy male SD rats, of 11 weeks old, weighing 320–370 g, were provided by Experimental Animal Faulty, Tongji Medical College, Huazhong University of Science and Technology. Rabbit-anti-phosphorylated ERK1/2 (pERK1/2) polyclonal antibody was provided by R&D Company; rabbit-anti-glial fibrillary acidic protein (GFAP) polyclonal antibody, SP immunohistochemical kit and horseradish peroxidase (HRP)-labeled goat-anti-rabbit IgG by Santa Cruz Company; specific inhibitor U0126 of ERK1/2 signal pathway by Alexis Company. METHODS: The experiment was carried out in the Laboratory of Neurosurgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from September 2004 to March 2006. ① Detection of pERK1/2 expression: A total of 110 rats were randomly divided into sham operation group (n =5), model group (n =35), high-dosage U0126 group (n =35) and low-dosage U0126 group (n =35). Rats in the sham operation group were only treated with incision of epicranium and fixation of backup plate, but not hit. Rats in the model group were used to establish diffuse brain injury models based on Marmarou free falling body without drug intervention. Rats in the high- and low-dosage U0126 groups were injected into caudal vein with 0.1 and 0.05 mg/kg U0126, respectively, and then, rats were hit to establish injured models. Every 5 rats were collected from model, high- and low-dosage U0126 groups at 5, 30 minutes, 3, 12, 24, 72 hours and 7 days after diffuse brain injury to detect pERK1/2 expression in cortex of parietal lobe based on Western blot technique. ② Distribution of pERK1/2 and positive GFAP cells in brain tissue: Another 48 rats were randomly divided into sham operation group (n =3), model group (n =15), high-dosage U0126 group (n =15) and low-dosage U0126 group (n =15). The intervention and administration were dealt as the same as those mentioned above. Every 3 rats were collected from model, high- and low-dosage U0126 groups at 30 minutes, 3, 12, 24 and 72 hours after model establishment to observe the distribution of pERK1/2 and postive GFAP cells in brain tissue which was cut from coronal section at Bregma –4.8 mm layer with immunohistochemical staining. MAIN OUTCOME MEASURES: pERK1/2 expression in cortex of parietal lobe and distribution of pERK1/2 and positive GFAP cells in brain tissues. RESULTS: ① pERK1/2 expression: After diffuse brain injury, pERK1/2 expression in cortex of parietal lobe was rapidly increased in the model group, reached at peak at 5 minutes and then decreased gradually. But the expression was still in a high level until the 72nd hour and fallen to the basic level on the 7th day. pERK1/2 level was lower in high- and low-dosage U0126 groups than that in model group at various time points (P < 0.01); meanwhile, pERK1/2 level was lower in high-dosage U0126 group than that in low-dosage U0126 group. The results showed that there was a certain dosage dependence on pERK1/2 expression. ② Distribution of pERK1/2 and positive GFAP cells in brain tissue: Positive expression of pERK1/2 lasted in brain tissue from 30 minutes to 72 hours after diffuse brain injury (P < 0.05). In addition, from 30 minutes to 3 hours, brown-yellow stained cells were mainly distributed in plasma, but rarely in nucleus. A lot of positive cells had tree-like apophysis, which was similar to neurons. With the time passing by, more and more nuclei manifested positive stains; moreover, nuclei mainly manifested positive staining until 24 hours after diffuse brain injury. Immune-positive pERK1/2 cells were widely distributed in brain tissue, especially mainly in binding site between deep cortex and cerebral white matter, and then in hippocampus. In addition, ependymal cell and vascular endothelial cells of choroids plexus also manifested strongly positive staining. As compared with model group, positive cells were decreased gradually in high- and low-dosage U0126 groups. However, number of positive cells was less in high-dosage U0126 group than that in low-dosage U0126 group. CONCLUSION: Diffuse brain injury strongly induces the activity of ERK1/2 signal pathway and response of astrocyte; in addition, U0126 can inhibit response of glial cells during an acute period, and the effect manifests dosage dependence.     

三嵌段高分子骨组织工程支架材料的体内生物学性能评价

目的：对三嵌段高分子骨组织工程支架材料-聚（丙交酯-乙交酯）[天冬氨酸-聚乙二醇1的生物相容性进行评价，探讨用于骨组织工程的可行性。 方法：实验于2003-08／12在华中科技大学同济医学院附属协和医院骨科实验室完成。①过敏试验：20只豚鼠随机分为试验组、对照组，每只豚鼠脊柱两侧皮内注射等体积聚（丙交酯-乙交酯）[天冬氨酸-聚乙二醇]浸提液、生理盐水及体积分数为0．05的甲醛溶液，记录激发部位红斑水肿。②急性全身毒性试验：12只小白鼠随机分为试验组和对照组，该两组腹腔分别注射浸提液及生理盐水，4，24，48及72h观察动物的一般状态。③植入试验：聚（丙交酯-乙交酯）[天冬氨酸-聚乙二醇]植入6只新西兰大白兔肌肉内，1，4及8周切取材料周围0．5cm处肌肉组织，苏木精-伊红染色，光镜下观察。 结果：①过敏试验结果：皮内注射浸提液及生理盐水动物皮肤无红斑水肿，注射体积分数为0．05的甲醛溶液出现中度以上红斑水肿。②急性全身毒性试验结果：阴性对照组未见毒性症状，试验组4h后仅1只有轻微的运动减少，24h后恢复正常。③植入试验结果：所有试验动物伤口均一期愈合。 结论：聚（丙交酯-乙交酯）[天冬氨酸-聚乙二醇]具有良好生物相容性，是一种新的骨组织工程支架材料。     

老年患者尿路感染菌群分布及其耐药性分析

目的 :了解老年患者尿路感染致病菌的菌群分布及其对抗生素的耐药情况 ,为临床合理使用抗生素提供依据。方法 :收集湖北省 15所三级甲等医院 2 0 0 2年尿路感染老年患者清洁中段尿细菌培养分离的 5 34株致病菌 ,对其进行耐药性监测。药敏采用K B法 ,用WHONET 5软件进行数据分析。结果 :共收集致病菌 5 34株 ,其中革兰阴性菌 4 0 9株 (76 .6 % ) ,革兰阳性菌 12 5株(2 3.4 % )。革兰阴性菌中大肠埃希菌检出率最高 (2 6 4株 ,4 9.4 % ) ,其次为克雷白杆菌 (44株 ,8.2 % )。 16 .7%的大肠埃希菌和 2 2 .7%的克雷白杆菌产超广谱 β 内酰胺酶。亚胺培南、阿米卡星、头孢他啶对革兰阴性菌的抗菌活性最强 ,而革兰阴性菌对环丙沙星、庆大霉素、哌拉西林的耐药率均在 5 0 %以上。革兰阳性菌以肠球菌最多见 (6 4株 ,12 % ) ,其次为葡萄球菌属 (43株 ,8.1% )。革兰阳性菌对SMZco、红霉素等的耐药率均在 4 0 %以上 ,但对万古霉素均敏感。结论 :老年患者尿路感染以革兰阴性菌为优势菌株 ,且耐药性日益严重 ,对亚胺培南、阿米卡星、头孢他啶最为敏感。革兰阳性菌宜以万古霉素为首选。     

Effects of basic fibroblast growth factor on biological characteristics of osteoblasts

Objective: To elucidate the effects of exogenous basic fibroblast growth factor ( bFGF ) on biological characteristics of rat osteoblasts cultured in vitro. Methods: The osteoblasts isolated from a Sprague-Dawley rat and cultured in vitro were treated with different concentrations of bFGF ( 5-50 ng/ml) respectively. At 24 hours after treatment, the proliferating cell nuclear antigen was measured with immunocytochemistry, alkaline phosphatase (ALP) activity was determined and the expression of transforming growth factor beta 1 ( TGF-β1 )was detected to observe the effects of bFGF on growth and differentiation of osteoblasts. Resu/ts: bFGF ( 5-50 ng/ml ) could obviously promote the growth of osteoblasts. The intracellular expression of TGF-β1 mRNA increased significantly, but the intracellular ALP content decreased. Conclusions: bFGF can obviously stimulate the proliferation of osteoblasts and promote the synthesis of TGF-β1, but cannot promote the differentiation of osteoblasts.     

维生素C浸泡羊膜对眼碱烧伤后移植的可用性研究

目的探讨维生素C浸泡羊膜的特性和生物活性物质的改变,以求为维生素C浸泡羊膜在眼碱烧伤羊膜移植中的合理应用提供依据。方法维生素C浸泡新鲜羊膜、保存1月羊膜,分别观察其大体物理特性、光学显微镜观察组织特性、免疫组织化学检测细胞因子的表达。结果维生素C浸泡羊膜与非浸泡羊膜的大体物理特性及光学显微镜观察组织特性无显著差别、部分细胞因子的活性表达无改变。结论维生素C浸泡羊膜不改变羊膜的组织学特性及部分细胞因子的活性,可以方便、安全地应用眼碱烧伤的移植治疗。     

核因子-κB在蛋白激酶C对支气管哮喘T淋巴细胞增殖和凋亡调控信号转导中的作用

目的　探讨核因子 κB(NF κB)在蛋白激酶C(PKC)对支气管哮喘T淋巴细胞增殖和凋亡调控的信号转导中的作用。方法　哮喘组和正常对照组的豚鼠以及急性发作期哮喘患者和正常对照者的外周血中分离出T淋巴细胞并分别加入PKC激动剂 12 肉豆蔻酰 13 乙酸佛波酯 (PMA)和NF κB抑制剂二硫代氨基甲酸吡咯烷 (PDTC)培养。用免疫组化法检测NF κB的表达 ,用四甲基偶氮唑盐微量比色法测定增殖反应 ,用原位末端终止法观测凋亡。结果　加入PMA培养的哮喘T淋巴细胞NF κB活化细胞百分比和增殖率与空白对照及加入PMA培养的正常T淋巴细胞相比均显著升高 (P <0 .0 1) ,而加入PDTC后以上指标均显著降低 (P <0 .0 1)。加入PMA培养的哮喘T淋巴细胞的凋亡指数与空白对照及加入PMA培养的正常T淋巴细胞相比均显著降低 (P <0 .0 1) ,加入PDTC后以上指标均显著升高 (P <0 .0 1)。T淋巴细胞NF κB活化细胞的百分比与增殖率均呈显著正相关 (r =0 .5 1～ 0 .72 ,P <0 .0 0 1) ,与凋亡指数均呈显著负相关 (r= 0 .5 5～ 0 .71,P <0 .0 0 1)。结论　T淋巴细胞PKC活化后导致增殖增加及凋亡减少的生物信号可能是通过激活NF κB来转导的。T淋巴细胞PKC NF κB信号转导途径的激活可能是哮喘的发病机制之一