光纤药物溶出度实时测定仪监测甲氨蝶呤片的溶出度

目的建立考察甲氨蝶呤片溶出过程的方法,初步评价制剂质量。方法用光纤溶出度实时测定仪实时监测甲氨蝶呤片的溶出过程。结果考察1个厂家10批产品的溶出曲线,结果显示样品批间差异和批内差异均很大,说明该厂家生产工艺不稳定。结论光纤溶出度实时测定仪能够有效测定固体药物的体外溶出过程,为改进制剂工艺、监控制剂工艺稳定性提供有益的参考。     

变性胶原体外培养模型的建立

目的 探讨不同温度对Ⅰ型胶原分子二级结构的影响,确定合适的胶原变性温度,研究热变性后胶原纤维排列及三维凝胶性质的改变,比较胶原变性后不同培养环境成纤维细胞形态差异,以建立变性胶原-细胞体外培养模型. 方法 Ⅰ型胶原蛋白溶液在不同温度作用后通过蛋白质圆二色光谱仪分析胶原分子二级结构改变.扫描探针显微镜观察胶原变性后纤维结构的改变.制备不同种类三维胶原凝胶并通过气相压力仪检测胶原凝胶断裂模量.将变性后的胶原进行二维包被和三维胶原凝胶制作,倒置相差显微镜及光镜下观察不同培养环境下细胞形态变化. 结果 温度达到50℃时,Ⅰ型胶原分子二级结构发生明显改变,在二维胶原包被时可见胶原纤维凝集成团,含变性胶原的三维凝胶断裂模量明显下降.在变性胶原存在环境中培养成纤维细胞,细胞形态均有显著改变. 结论 经50℃作用后Ⅰ型胶原分子二级结构发生明显改变,含变性胶原的三维凝胶断裂模量明显下降,Ⅰ型胶原包被及三维凝胶模型培养的成纤维细胞形态明显不同,可作为变性胶原影响细胞生物学活性的体外模型.     

大黄水提物不同条件干燥产物理化性质差异的初步研究

目的考察大黄水提液不同条件下干燥产品的理化差异性。方法采用傅里叶变换红外光谱法(FTIR),对不同样品进行分析;并在红外分析结果基础上,利用HPLC法对样品中的5种蒽醌成分进行含量测定,分析FTIR结果与样品化学成分差异的相关性。结果冷冻干燥样品与其他两种干燥工艺样品红外图谱和蒽醌含量差别明显。而喷雾干燥和微波干燥不同条件干燥样品FTIR图谱总体差异不明显,但通过聚类分析将其样品分为3大类。含测结果亦表明第3类中6号(喷雾干燥:进/出风温205~215℃/105~110℃)和10号(微波干燥:10Kw)样品5种蒽醌类成分含量较其他样品差异较大。结论大黄水提液的喷雾和微波干燥工艺样品同冷冻干燥样品有显著差异在一定条件下工艺稳定,适合制剂工业生产。但喷雾干燥工艺进/出风温不宜超过205~215℃/105~110℃,而微波干燥功率应低于10Kw。     

高压氧干预下急性脑梗死患者细胞黏附分子变化及对预后的影响

目的 对急性脑梗死(acute cerebral infarction,ACI)患者CD11b、CD11c和CD54作动态观察,找出其变化规律及高压氧(hyperbaric oxygen,HBO)对该规律的影响,并进一步阐明HBO对ACI患者脑血管的保护作用;同时对ACI患者治疗前CD11b、CD11c和CD54与近、远期神经功能恢复程度相关性进行探讨.方法 64例ACI患者分2组:HBO组31例,常规治疗联合HBO治疗;治疗对照组33例,仅行常规治疗.2组分别在治疗前(发病≤72 h)及治疗第7、10、12、20天晨抽取外周静脉血测定CD11b、CD11c和CD54,并在相同治疗时间进行神经功能缺损程度评分(neural functional damage scores,NDS).另外选取正常对照组25人,仅抽取周围静脉血测定CD11b、CD11c和CD54.结果 HBO组和治疗对照组治疗前CD11b、CD11c和CD54表达明显升高,达到高峰,2组各指标差异无统计学意义(P＞0.05),第7天均开始下降.HBO组CD11b、CD11c高峰持续时间为7d,治疗对照组持续10 d;HBO组CD54高峰持续时间为10 d,治疗对照组持续12d.经相关性和线性回归分析发现,发病第20天时NDS水平(近期疗效)和发病半年、1年时NDS的水平(远期疗效)分别为97.2％、96.7％及97.6％,可用CD11b和/或CD11c和/或CD54的上调水平以及其他相关因素解释.结论 HBO治疗后可缩短ACI患者CD11b、CD11c和CD54高峰持续时间.说明HBO可干预细胞黏附分子的变化过程,缩短其异常时限,减少它们的表达,对ACI患者有保护作用,并减少白细胞的黏附.同时治疗前CD11b、CD11c和CD54的表达可预测病情的轻重,影响近、远期疗效,为研究如何调控或抑制CD11b、CD11c和CD54的表达、减少对血管的损伤及早期制定更全面的治疗方案提供依据.     

G-CSF对健康人骨髓和外周血CD34+细胞和T细胞亚群影响的动态观察

目的 :探讨G -CSF对健康人骨髓及外周血CD3 4 + 细胞和T细胞亚群的影响。方法 :对 12例健康人 ,以G -CSF15 0 μg/ 12h ,皮下注射 ,连用 6d。应用流式细胞仪测定CD3 4 + 和T细胞亚群。结果 :应用G -CSF4 8h后 ,骨髓CD3 4 + 细胞开始明显增加 ,72h后外周血CD3 4 + 细胞开始明显增加 ,96h后两者达峰值 ,均较应用前差异显著 (P <0 .0 1)。T淋巴细胞亚群变化不明显 (P>0 .0 5 )。结论 :健康人应用G -CSF后骨髓及外周血CD3 4 + 细胞逐渐增加并于 96h后达到高峰 ,T细胞亚群则无明显变化。     

高压氧对SARS后股骨头缺血坏死患者血小板活性相关指标的影响

目的　探讨高压氧 (HBO)对传染性非典型肺炎 (SARS)后股骨头缺血坏死患者血小板膜糖蛋白CD31,CD61,CD62p,CD63和PAC 1的影响。方法　治疗组 26例SARS后股骨头坏死患者HBO治疗前后取静脉血,测定血小板膜糖蛋白CD31,CD61,CD62p,CD63和PAC 1的表达并分别与17例正常人对照组进行对照。结果　治疗组血小板膜糖蛋白CD31,CD61,CD62p,CD63和PAC 1HBO治疗前均低于对照组,与对照组比较有差异显著性 (P<0. 01 ),HBO治疗后均升高,与治疗前比较差异有统计学意义 (P<0. 01或P<0. 05),而治疗后与对照组比较差异无统计学意义 (P>0. 05 )。结论　HBO治疗可以使SARS后股骨头缺血坏死患者血小板膜糖蛋白CD31,CD61,CD62p,CD63和PAC 1的表达恢复正常。     

核黄素磷酸钠粉针处方及制备工艺研究

目的 通过试验确定核黄素磷酸钠粉针剂的处方和制备工艺.方法 通过对冻干品的外观、含量、含水量、复溶性和溶解后澄明度等指标的考察确定最佳的支撑剂种类和用量、药液处理方法和冻干工艺参数.结果 冻干前处方组成为核黄素磷酸钠13.69 g、焦亚硫酸钠2.0g、EDTA-2Na 1.0 g、溶解于2000 ml蒸馏水中,室温下加...     

抗CD25单克隆抗体对肾移植受者CD4~+ CD25~(high)调节性T细胞的影响

目的 评价抗CD25单克隆抗体对肾移植受者术后早期CD4~+ CD25~(high)调节性T细胞(CD4~+ CD25~(high)Treg)的影响.方法 2007年2-9月接受初次亲属活体供肾移植的受者41例,根据是否使用抗CD25单克隆抗体(商品名daclizumab)分为抗体组(21例)和对照组(20例).其中抗体组在肾移植术前2h及术后第14天分别给予抗CD25单抗各50mg.在移植前及移植后第13、17、60天分别留取肝素抗凝外周血15ml.应用流式细胞仪测定两组受者外周血CD4~+T细胞和CD4~+ CD25~(high)Treg比例的变化,半定量RTPCR检测CD25 mRNA的表达变化.结果 肾移植术后13、17、60d抗体组的CD25~+ T细胞占CD4~+ T细胞的比例(20%±8%、13%±7%、24%±9%)低于对照组(45%±6%、41%±5%、40%±6%),差异有统计学意义(P<0.05).抗体组术后第17天CD4~+ CD25~(high)Treg占CD4~+ T细胞的比例为4.40%±0.26%,明显低于对照组(8.56%±0.36%,P<0.01);而术后13、60d抗体组CD4~+ CD25~(high)Treg所占比例分别为7.00%±0.47%、3.75%±0.19%,与对照组(分别为8.04%±0.32%、3.66%±0.31%)比较差异无统计学意义(P>0.05).抗体组CD25 mRNA相对表达水平在给予第二次抗体前(术后第13天)为1.65±0.22,术后第17天为1.84±0.27,两者间差异无统计学意义(P>0.05).对照组术后第17天CD25 mRNA相对表达水平为1.70±0.23,与抗体组比较差异无统计学意义(P>0.05).结论 两剂共100mg抗CD25单抗仅一过性地降低CD4~+ CD25~(high)Treg,不会影响其活化扩增,无损于术后早期的免疫耐受诱导及维持.     

肝移植后患者外周血Tfh细胞亚群的变化特征及意义

目的 探讨肝移植术前和术后患者外周血滤泡辅助性T细胞(Tfh)亚群的表达变化及其与预后的相关性.方法 对11例肝移植术后肝功能稳定的患者进行随访研究,采用流式细胞术检测移植前和移植后4周外周血中Tfh细胞亚群频率的变化.结果 与移植术前比较,移植术后4周患者外周血Tfh1 (CD4+CXCR5+CXCR3+CCR6)细胞频率较移植前有下降趋势,但差异无统计学意义(P＞0.05),Tfh2(CD4+CXCR5+CXCR3-CCR6)细胞频率较移植前明显升高(P＜0.05),Tfh1 7(CD4+CXCR5+CXCR3-CCR6+)细胞频率较移植前有上升趋势,但差异无统计学意义(P＞0.05).结论 Tfh2和Tfh17细胞可能与肝移植后患者肝功能的恢复有关.     

肝移植患者外周血Tfh细胞特征的初步研究

目的检测肝移植术前和术后患者外周血滤泡性辅助性T细胞(Tfh)及其分子表达特点,探讨其在肝移植免疫中的作用。方法肝移植术后患者及健康对照各20例,其中移植术后肝功能稳定者13例,对其进行断面研究;余7例病例资料完整,进行随访研究。应用流式细胞仪检测所有入组者的Tfh细胞频率,并对随访患者进行术后1月的动态监测,同时检测其丙氨酸转氨酶(ALT)水平。结果与移植术前相比,移植术后患者外周血CD4+CXCR5+Tfh和CD4+CXCR5+PD-1+细胞频率明显升高(P<0.05),CD4+CXCR5+ICOS+细胞虽有上升趋势,但无统计学意义。随访发现移植术后肝功能稳定患者CD4+CXCR5+Tfh水平在移植术后明显上升,1周后达峰值随后下降,其ALT水平也在术后第10天达到峰值后降低。随访期间1例患者发生急性排斥反应,其CD4+CXCR5+Tfh水平与ALT水平变化趋势一致。肝功能稳定组与健康对照组相比,CD4+CXCR5+Tfh、CD4+CXCR5+PD-1+和CD4+CXCR5+ICOS+细胞频率均明显升高(P<0.05)。结论外周血CD4+CXCR5+Tfh细胞在肝移植后表达上调,可能参与了肝移植术后的免疫调节。     

动态浊度法测定注射用重组人钙调磷酸酶B亚基中的细菌内毒素

目的对注射用重组人钙调磷酸酶B亚基(rhCNB)进行细菌内毒素定量回收试验，建立定量检测其细菌内毒素的方法。方法采用中国药典2000年版附录检测细菌内毒素的动态浊度法。结果注射用rhCNB在20倍稀释后可以消除干扰作用，内毒素回收率在50％～200％范围内，能够进行细菌内毒素定量检测。结论建立的注射用rhCNB细菌内毒素定量检查方法，具有方法简便、快速、准确的优点。     

穿膜肽增强聚乙烯亚胺介导的基因转染效率的机制研究

目的研究穿膜肽增强支链聚乙烯亚胺（BPEI）在CHO-K1细胞中的基因转染效率的机制。方法分别合成蜂毒肽（melittin）及其疏水核心区MT20,利用圆二色光谱（CD）分析其二级结构;通过溶血试验比较二者穿透细胞膜的能力;加入蜂毒肽和MT20前后,观察钙黄绿素（calcein）在HeLa细胞内的分布情况。分别将蜂毒肽或MT20与BPEI按一定比例混合,以荧光素酶（luciferase）为报告基因,检测荧光素酶在CHO-K1细胞中的转染效率;以MTT法检测基因转染后的细胞毒性。结果在模拟膜环境的甲醇溶液中,蜂毒肽和MT20都呈现一定的α-螺旋结构,比例分别为59.63%和35.67%,说明其二级结构与之穿膜功能相关;蜂毒肽只能在中性条件下穿透细胞膜导致红细胞溶血,而MT20在中性和酸性条件下都具有穿膜能力;加入蜂毒肽和MT20后能够促进钙黄绿素在MT20组中呈弥散分布,在蜂毒肽组中呈颗粒状分布并且在核仁内蓄积;MT20和蜂毒肽都能够增强BPEI在CHO-K1细胞中的基因转染效率,其中MT20的增强作用更强且细胞毒性较蜂毒肽以及单独使用BPEI和Lipofectamine 2000时要低。结论蜂毒肽的疏水核心区MT20通过增加PEI/DNA复合物颗粒从内含体逃逸并促进其进入细胞核而增加其基因转染效率,是一种低毒的基因转染增强剂,有可能在难以转染的细胞系中发挥重要作用。     

Preparation, analysis and application of [99mTc]Human Albumin Microspheres ([99mTc]HAM) for lung scanning

Kits were developed for the sterile labelling of Human Albumin Microspheres with 99mTc. The microspheres were prepared, sieved to get the desired particle size and autoclaved for sterilization. The spheres were treated with stannous chloride and the pH of the suspension was adjusted to 3.7 with phosphate buffer. After freeze-drying the contents of single reaction vials from different batches were reacted with 99mTc and the radiochemical yield (higher than 97%) was determined. The HAM kits were stable and the stability of [99mTc]HAM was followed for 5 h. Lung and liver uptake in mice was determined to be about 90 and 1% respectively. The preparation of [99mTc]HAM is performed in a single step process and excellent human lung scans were obtained.     

平行秋千摆动刺激中的心率变异分析

目的 观察飞行员在平行秋千摆动前、中、后反映心脏植物神经调节功能的心率变异性（ＨＲＶ）特点,搪塞以此评价线性加速度作用下前庭＝植物神经反应性的可能性。方法记录３７名飞行员在秋千摆动前、中、后的连续心电图,进行频率谱分析并计算低频成分（ＬＦ）和高频成分（ＨＦ）之幽会对被试者中ＬＦ／ＨＦ值在不同摆动阶段变化类型及分布耐受摆动时间的关系进行初步分析。结果 摆动中的ＬＦ明显降低,ＨＦ明显增高,ＬＦ／ＨＪＦ     

糖尿病犬经肝动脉肝内移植微囊化猪胰岛细胞的安全性和有效性

目的评估移植于I型糖尿病犬肝脏中的微囊化新生猪胰岛细胞(NPI)的生物相容性、免疫学特性及生理学特性。方法I型糖尿病犬分为A、B两组,每组15只,A组每只犬分别经肝动脉灌注微囊化新生猪胰岛细胞40～60万个,B组每只犬分别经肝动脉灌注未微囊化新生猪胰岛细胞40～60万个,两组动物移植后均不使用免疫抑制治疗。移植前后分别测量移植受体的肝脏功能及淋巴细胞CD4 /CD8 比值。移植6个月后所有移植受体的肝脏均进行病理学检查。结果移植后A组外源性胰岛素用量从移植前的22 u逐渐降至5 u,B组所需外源性胰岛素从移植前的24 u下降至10 u。移植后2～3周B组胰岛素用量恢复到移植前的水平,而A组的部分动物的胰岛素剂量继续减至8 u。B组受体移植后血的CD4 较移植前升高,而A组的CD4 和CD8 细胞移植后无明显变化。移植后所有受体的肝功能及组织结构未见异常。结论微囊化的新生猪胰岛细胞在受体犬的肝脏中有很好的生物相容性。微囊可以延长移植物的存活,且异种移植微囊化的新生猪胰岛细胞能够纠正糖尿病犬的高血糖状态。     

DNA fingerprinting of freeze-dried tissues

Summary DNA profiling is usually unproblematic when carried out on material sampled shortly after death. Prolonged postmortem intervals and improper storage cause significant DNA degradation, however there are instances where DNA analysis becomes necessary months after the autopsy. In such cases the investigator may be able to revert to material initially stored for toxicological purposes. In some cases such specimens undergo freeze-drying before storage. We have therefore tested DNA fingerprinting of freeze-dried postmortem material. It was found that freeze-drying is a suitable method for preserving tissue samples for DNA profiling.Some contents of this paper were communicated as a poster at the 14th Congress of the International Society of Forensic Haemogenetics (Mainz, September 18–21, 1991)     

Short-term changes in the immune system of elite swimmers under competition conditions: Different immunomodulation induced by various types of sport

This study was designed to investigate changes in the immune system of elite swimmers compared with well-conditioned age- and sex-matched controls in relation to a competition swim (field study). Furthermore, the aim was to reveal possible differences in immune system changes depending on the type of sport performed by comparing with an earlier study of similar design, from the same laboratory that tested elite runners in relation to a competition run. The swimmers were tested before, immediately after and 2 h and 24 h after a competition swim. Lymphocyte subsets (CD5, CD3, HLA-DR, CD4, CD8, CD19, CD3/CD16+56, CD57, CD18, CD16/CD122) all increased after the run, decreased to normal or subnormal levels after 2h, and returned to normal after 24 h (absolute numbers). The findings were identical for the swimmers and the age- and sex-matched control group. No change in polymorphonuclear granulocyte migration was found. The lymphocyte proliferative responses decreased 2 h after the exercise. No changes were seen in plasma cytokine levels (interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) in relation to exercise, but significantly lower baseline values for IL-6 were observed in the swimmers. An increase in total natural killer cell activity immediately after exercise, followed after 2 h by a decrease, was seen in both swimmers and controls. Finally, no complement activation was detected. Compared with an earlier study of elite runners, differences were seen in granulocyte chemotactic response, TNF-α plasma activity and the lymphocyte proliferative response to mitogen. These differences might be explained by the degree of immune system activation following muscle damage during exercise, inducing an increase in cytokines, which are known to activate and modulate both lymphocytes and granulocyte function. Our findings demonstrate identical exercise-induced, immune system changes in elite swimmers and well conditioned controls, and furthermore, the findings suggest that different types of sport performed at maximum intensity induce different immune system changes.     

18F-脱氧葡萄糖注射液质量研究

目的 考察国内医疗机构制备的18F-脱氧葡萄糖(FDG)注射液的质量状况,并验证国家食品药品监督管理局标准YBH05262005(试行)用于不同医疗机构、采用不同工艺制备的18F-FDG注射液质量控制的适用性.方法 依据国家食品药品监督管理局标准YBH05262005(试行)对不同医疗机构、采用不同生产工艺制备的33批18F-FDG注射液样品进行全项检验;对国家食品药品监督管理局标准YBH05262005(试行)中的部分项目进行方法学验证实验;任选其中16批样品,按顶空进样毛细管气相色谱法进行有机溶剂残留量检验,并进行了方法学研究.结果 (1)33批18F-FDG注射液样品中,6批样品的检验结果不符合国家食品药品监督管理局标准YBH05262005(试行)规定.(2)方法学验证实验结果表明,国家食品药品监督管理局标准YBH05262005(试行)"[鉴别](3)"项规定的18F-FDG Rf值范围(0.6～0.7)与实验结果(0.9～1.0)有一定差异,其他项目方法适于注射液的质量控制.(3)方法学研究结果表明,顶空进样毛细管气相色谱法适于样品有机残留最的检查,16批样品检验结果中有1批样品的乙腈残留量(0.510 64%)超出中国约典(2005年版)二部附录(Ⅷ P)和美国药典(31版)规定的限度,乙醚和乙醇残留量结果均符合上述中国和美国药典规定.结论 为确保18F-FDG注射液用药安全有效,其制备医疗机构需进一步加强制备和质最管理;国家食品药品临督管理局标准YBH05262005(试行)也需进一步完善,如修订"[鉴别](3)",并增订有机溶剂残留量检查项等.     

LeuCAM and reactive oxygen species during long-term exercise combined with sleep and energy deficiency

INTRODUCTION: During the Norwegian military ranger-training course, cadets are exposed to prolonged physical exercise combined with sleep-, energy-, and food deficiency. The open-window postexercise hypothesis indicates that after hard physical activity, there is an increased risk of contracting infectious diseases. PURPOSE: The purpose of the present study was to determine leukocyte reactive oxygen species (ROS) levels, total antioxidant status (TAS), leukocyte expression of the cell adhesion molecules CD62L and CD11b, and plasma levels of soluble adhesion molecule L-selectin before, during, and in the recovery phase of a military ranger-training course. METHODS: Ten cadets from the Norwegian Military Academy were recruited to the study. Flow cytometry was used to study the intracellular levels of ROS in leukocytes (basally, as well as after in vitro stimulation with phorbol myristate acetate (PMA)), applying the probes dihydroethidium (DHE) and dihydrorhodamine 123 (DHR) and the leukocyte expression of adhesion molecules CD62L and CD11b. ELISA was used to assess the plasma levels of soluble L-selectin, and TAS in plasma was measured using the ABTS+ reduction assay kit. RESULTS: The basal levels of ROS as well as PMA-stimulated ROS in leukocytes declined gradually during the ranger-training course, being lowest on the last day (P < 0.05). The level of TAS increased (P < 0.01) during the course. A striking decrease (P < 0.001) was observed in leukocyte CD62L expression and was sustained even after 3 d of recovery. The leukocyte expression of CD11b remained unchanged. CONCLUSION: The ranger-training course leads to a partial exhaustion of the leukocyte ROS-generating machinery and to a nearly total extinguishing of leukocyte CD62L expression. These changes may support the open-window hypothesis indicating reduced ability to combat microbial invasions before total restitution.