糖尿病大鼠卵巢组织中细胞色素C、线粒体Ca2+变化与灵芝孢子粉的干预

目的:以卵巢组织中细胞色素C、线粒体Ca2 和性激素水平为观察指标,观察灵芝孢子粉对2型糖尿病大鼠卵巢组织的保护作用。方法:实验于2006-06/08在佳木斯大学黑龙江省小儿神经康复重点实验室完成。①实验对象:Wistar雌性大鼠50只,随机分为对照组10只,2型糖尿病模型组20只,灵芝孢子粉组20只。②实验方法:大鼠禁食16～18h,自由饮水,模型组及灵芝孢子粉组大鼠经尾静脉一次性按25mg/kg注射2%链脲佐菌素,正常对照组尾静脉注射等量柠檬酸钠-柠檬酸缓冲液。正常饮食喂养2周后,进行糖耐量试验,模型组和灵芝组以糖耐量异常者保留,余去除,并改喂高脂高糖饮食,灵芝孢子粉组另加灵芝孢子粉[250mg/(kg·d)]持续10周,实验结束前一天再次做糖耐量试验。③实验评估:实验结束后,各组动物均禁食12h,乙醚吸入浅麻醉,内眦静脉取血待测血清胰岛素。解剖出卵巢,待测线粒体细胞色素C、线粒体Ca2 含量。结果:选用Wistar雌性大鼠50只,结果分析数量每组8只。①葡萄糖耐量试验:实验开始及实验12周时,模型组大鼠均表现出糖耐量异常的变化(P<0.05),1h血糖增高显著(P<0.05)达到高峰,2h血糖仍较高;对照组大鼠30min血糖达到峰值,2h血糖基本恢复正常。②血清胰岛素和卵巢性激素水平:模型组血清胰岛素明显升高(P<0.05)。而灵芝孢子粉组与对照组比较差异无显著性。模型组雌二醇含量明显降低(P<0.05),卵胞刺激素含量、睾酮含量明显升高(P<0.05),而灵芝孢子粉组与对照组比较差异无显著性。③胞浆细胞色素C、线粒体细胞色素C、线粒体钙变化:模型组卵巢组织胞浆细胞色素C含量明显升高(P<0.05),模型组卵巢组织线粒体细胞色素C含量明显降低(P<0.05),模型组卵巢组织线粒体钙含量明显降低(P<0.05);而灵芝孢子粉组与对照组比较差异均无显著性。结论:灵芝孢子粉可降低糖尿病大鼠血清胰岛素、卵巢组织卵胞刺激素、睾酮、胞浆细胞色素C含量,调节卵巢组织雌二醇、线粒体细胞色素C、钙含量,对糖尿病大鼠卵巢组织具有一定保护作用。     

Study on the in vitro and in vivo activation of rat hepatic stellate cells by Raman spectroscopy

The feasibility of a novel and efficient diagnostic method for liver fibrosis using Raman spectroscopy is studied. Confocal Raman spectroscopy (CRS) is utilized to monitor the molecular changes of hepatic stellate cells (HSCs) in vitro as well as in vivo activation. In vitro activation was induced by growth in uncoated plastic plates, while the in vivo activation is induced by a single intraperitoneal injection of carbon tetrachloride (CCl(4)). The biochemical changes of HSCs during activation such as the loss of retinoid, the increase of alpha-helical protein, and the increased production of extracellular matrix proteins are observed by CRS. A user-friendly autoclassifying system is also developed to classify Raman spectra of liver injury tissues with a 90% accuracy rate. Raman spectroscopy combined with a fiber optical probe could be potentially accomplished for in vivo detection, which can lead to a novel and efficient diagnosis for liver fibrosis.     

射频皱缩刀在不同工作模式下对腋神经表面温度的影响

目的：观察肩关节镜下射频皱缩刀对腋神经表面温度的影响. 方法：实验于2005-03/06在南京医科大学附属南京第一医院骨科实验室进行.取10个新鲜肩关节标本（南京中医药大学提供）,分为肩关节持续冲洗和间断冲洗两组,模拟肩关节不稳定进行射频皱缩刀关节囊皱缩术.射频皱缩刀应用持续工作和间断工作两种不同的工作模式和1和2档能量设置,采用电动测温仪分别记录射频皱缩刀工作10,25,40,55,70 s及停止工作30 s时腋神经表面的温度. 结果：①肩关节间断冲洗组：射频皱缩刀持续工作模式下,各个时间段腋神经表面温度升高较快,2 min以后均超过70℃,最高达到85℃（3 min时）,停止工作30 s,仍为82.3℃;射频皱缩刀间断工作模式下腋神经表面温度较持续工作模式升高慢（P＜0.01）,各个时间点均不超过60℃,最高才达到58℃（70 s时）.②肩关节持续冲洗组：射频皱缩刀持续工作模式下,皱缩能量为1档和2档时,在30 s时腋神经表面的温度不差异（P＞0.05）,而连续工作1 min以上,能量为1档时,各个时间点所测温度均比能量为2档时低（P＜0.05）;射频皱缩刀间断工作模式下,皱缩能量为1档和2档时,在10,25 s时,腋神经表面温度差异不显著（P＞0.05）,40 s后,能量为1档时,腋神经表面温度比能量为2档时低（P＜0.05）.在肩关节持续冲洗和能量为1档时,射频皱缩刀在两种工作模式下,腋神经表面温度均不超过50℃. 结论：在肩关节镜下行射频皱缩术时,腋神经表面温度会明显升高,持续冲洗、间断工作模式可降低腋神经表面温度;而间断冲洗、持续工作模式可能会损伤腋神经.     

5,6-二甲氧基-1-吲酮的合成

目的：合成5,6－二甲氧基－1－吲酮（Ⅰ）,并对其合成工艺进行改进。方法：以3,4－二甲氧基苯甲醛为起始原料,经缩合、还原、环合等步骤制备而成。结果：三步反应的总收率达71．8％,合成产物经熔点和红外光谱确证。结论：此合成工艺简便,易于工业化生产。     

Differential kinetics of primitive hematopoietic cells assayed in vitro and in vivo during serum-free suspension culture of CD34+ blood progenitor cells.

So far, blood progenitor cells (BPC) expanded ex vivo in the absence of stromal cells have not been demonstrated to reconstitute hematopoiesis in myeloablated patients. To characterize the fate of early hematopoietic progenitor cells during ex vivo expansion in suspension culture, human CD34(+)-enriched BPC were cultured in serum-free medium in the presence of FLT3 ligand (FL), stem cell factor (SCF) and interleukin 3 (IL-3). Both CD34 surface expression levels and the percentage of CD34+ cells were continuously downregulated during the culture period. We observed an expansion of colony-forming units granulocyte-macrophage (CFU-GM) and BFU-E beginning on day 3 of culture, reaching an approximate 2-log increase by days 5 to 7. Limiting dilution analysis of primitive in vitro clonogenic progenitors was performed through a week 6 cobblestone-area-forming cell (CAFC) assay, which has previously been shown to detect long-term bone marrow culture-initiating cells (LTC-IC). A maintenance or a slight (threefold) increase of week 6 CAFC/LTC-IC was found after one week of culture. To analyze the presence of BPC mediating in vivo engraftment, expanded CD34+ cells were transplanted into preirradiated NOD/SCID mice at various time points. Only CD34+ cells cultured for up to four days successfully engrafted murine bone marrow with human cells expressing myeloid or lymphoid progenitor phenotypes. In contrast, five- and seven-day expanded human BPC did not detectably engraft NOD/SCID mice. When FL, SCF and IL-3-supplemented cultures were performed for seven days on fibronectin-coated plastic, or when IL-3 was replaced by thrombopoietin, colony forming cells and LTC-IC reached levels similar to those of control cultures, yet no human cell engraftment was recorded in the mice. Also, culture in U-bottom microplates resulting in locally increased CD34+ cell density had no positive effect on engraftment. These results indicate that during ex vivo expansion of human CD34+ cells, CFC and LTC-IC numbers do not correlate with the potential to repopulate NOD/SCID mice. Our results suggest that ex vivo expanded BPC should be cultured for limited time periods only, in order to preserve bone-marrow-repopulating hematopoietic stem cells.     

Genetic modulation of ADH1B and ALDH2 polymorphisms with regard to alcohol and tobacco consumption for younger aged esophageal squamous cell carcinoma diagnosis

Genetic variants in alcohol dehydrogenase‐1B (ADH1B) and aldehyde dehydrogenase‐2 (ALDH2) genes modulate acetaldehyde removal upon alcohol ingestion. Although these genetic vulnerabilities have been linked to higher esophageal squamous cell carcinoma (ESCC) risks, it is unclear whether they also determine the time of malignancy presentation. The purpose of this investigation was to unravel genotoxic effects of the two alcohol‐metabolizing genes with regard to alcohol and tobacco consumption on the age at ESCC diagnosis and tumor dissemination. ADH1B/ALDH2 genotyping was performed on lymphocyte DNA specimens taken from 406 consecutively registered incident patients with pathology‐proven ESCC. To fully utilize individual genetic and survival information, survival analyses and gene‐longevity applied approaches were introduced. Among heavy drinkers, the ADH1B Arg/Arg (55 years) and ALDH2 Glu/Lys genotypes (54 years) were found to confer a 15 and 16 years earlier carcinoma diagnosed age than His/His and Glu/Glu nondrinkers (both 70 years), respectively. For drinkers, 1‐year age advancement was, separately, associated with a 0.977 and 0.953‐fold stepwise reduced likelihood of being ADH1B Arg homozygote and ALDH2 Lys variant. Noticeably elevated hazard‐ratio (HR) for drinkers of ADH1B slow‐form genotype and ALDH2 inactive‐form allele were identified in smokers (HR = 2.3–2.6), but no in nonsmokers. In smokers, appreciably higher cumulative cancer onset risks were correspondingly recognized from the age of 45 and 49 upward among any + Lys allele and Arg/Arg + Glu/Glu combined‐ADH1B/ALDH2‐genotype drinkers than nondrinkers. In conclusion, consumption of tobacco and alcohol, coupled with genetic susceptibilities associated with acetaldehyde elimination, as modulated by ADH1B and ALDH2 genotypes, determines a substantial magnitude of tumorigenetic effect on earlier age ESCC diagnosis. © 2009 UICC     

Osteopontin Expression in Squamous Cell Cancer of the Esophagus

BACKGROUND: The purpose of this study was to better understand the role of osteopontin (OPN) in esophageal squamous cell carcinoma (ESCC) by comparing the OPN mRNA level in ESCC tumor tissue and matched normal tissue and by determining the prognostic significance of the gene expression. METHODS: Initially, by an oligo-nucleotide microarray hybridization technique, OPN expressions were found to consistently elevate at least twofold in three ESCC tissues compared with their adjacent normal ones. Subsequently, the expression of OPN mRNA was detected by real-time QRT-PCR among the 58 fresh surgical ESCC specimens. The clinical information was obtained by chart review. RESULTS: OPN mRNA expression was detectable in 58 of 58 (100%) tumor specimens and 57 of 58 (98.3%) nonmalignant esophageal specimens. OPN expression was higher in tumor tissue than in the matched normal tissue in 54 of 58 (93.1%) individual cases. The overall median mRNA expression level of OPN was approximately 8.8-fold higher in tumor tissues (4.1; range: 0.02-247) compared with matched normal esophageal tissues (0.5; range, 0.0-21.4; p < 0.001). Overexpression of OPN mRNA was significantly associated with clinical stage (p = 0.01). The more severe the clinical stage (from I-II, III, to IV) was the higher frequency of overexpression of OPN mRNA. No significant associations were found between overexpression of OPN mRNA and the patients' survival (p = 0.27). DISCUSSION: Our findings suggest OPN is associated with esophageal tumorigenesis and progression, but not patients' survival.     

瑞力芬片剂口服人体药代动力学及生物利用度研究

目的:检测2种萘丁美酮片剂的药代动力学及生物利用度是否有差异.方法:12例男性健康受试者交叉口服2种萘丁美酮片剂,用高效液相色谱法规定服药后120h内的血浆中蔡丁美酮活性代谢物6-甲氧基-2-萘乙酸浓度.结果:测得主要药代动力学参数为;试验片剂Tp=6.41±3.05h,t1/2β=21.77±3.62h,c_(max)=23.73±4.10mg/L,AUC=1.10.67±205.54(mg·h)/L;对照片剂Tp=10.63±5.10h,t1/2β=21.53±4.77h,c_(max)=18.77±5.39mg/L,AUC= 954.10±263.66(mg·h)/L.结论:试验片与对照片相比吸收较快(P=0.012),峰浓度较高(P=0.029),平均相对生物利用度为112.36%,AUC经配对T检验,无显著性差异(P=0.52).     

HDV/HBV感染树鼩肝组织ICE表达及肝细胞凋亡

目的 探讨HDV/HBV感染树鼩肝组织中ICE表达与HDV感染之间的关系,以及ICE在丁型肝炎肝细胞凋亡中的作用。 方法 采用免疫组化技术对45份HDV感染树鼩肝组织中HDVAg,ICE的表达进行了检测;应用原位末端标记技术对肝组织凋亡进行检测;并应用免疫组化双重染色对HDVAg,ICE的表达以及肝细胞凋亡进行了检测。 结果 肝组织45份中有43份可检出ICE(阳性率96％),有41份可检出凋亡细胞(阳性率91％),ICE在肝细胞浆和(或)膜上表达,以肝细胞浆内表达为主,HDVAg表达与ICE表达之间有显著相关性X~2=36.2(P<0.05),HDVAg表达越强,ICE表达越强,凋亡阳性细胞在肝细胞索多呈散在分布,在坏死灶中也可检出,凋亡在HDVAg阳性和阴性细胞中均可发生,以在HDVAg阳性细胞内发生为主,ICE表达与肝细胞凋亡之间有显著相关性X~2=35.6(P<0.05),ICE表达越强,凋亡阳性细胞越多。 结论 丁型肝炎病毒感染和未感染的肝细胞均可发生凋亡,但凋亡只在少数细胞内发生;HDV可诱导ICE表达,ICE在肝细胞凋亡中起重要作用。