健胃愈疡颗粒干预下大鼠胃溃疡黏膜乳癌相关肽和血小板活化因子的表达及与胃黏膜疏水性的相关性研究

目的:观察对胃溃疡复发有较好疗效的健胃愈疡颗粒对溃疡黏膜乳腺癌相关肽和血小板活化因子表达的影响,分析其可能的作用机制。方法:实验于2005-07/2006-07在湘雅医院中心实验室完成。SD大鼠110只,雌雄各半,随机抽签法分为5组,即正常对照组、假手术组、雷尼替丁组、健胃愈疡组,各20只;模型组30只。以Okabe改良法复制大鼠实验性胃溃疡,假手术组仅以生理盐水代替乙酸注入玻管内。造模后24h,雷尼替丁组、健胃愈疡组大鼠分别灌服盐酸雷尼替丁和健胃愈疡颗粒(药物组成为:柴胡、党参、白芍、延胡索、白芨、珍珠层粉、青黛、甘草,湖南湘雅制药有限公司生产)药液10mL/kg,分别相当于2.70,1.62g/kg,1次/d。假手术组、模型组灌服蒸馏水10mL/kg。10d后各组中随机取出10只大鼠剖腹取胃(处死前大鼠禁食24h),90d时将模型组20只大鼠再分为模型复发组和模型非复发组,各10只;除正常对照组、假手术组、模型非复发组大鼠腹腔内注射生理盐水外,其余各组大鼠腹腔内注射白细胞介素1,1μg/kg;在注射48h,大鼠禁食24h后,剖腹取胃。观察其对胃溃疡大鼠胃黏膜氨基己糖及磷脂含量、溃疡指数和胃黏膜血流的影响,并用RT-PCR观察乳癌相关肽乳癌相关肽和血小板活化因子表达的变化。结果:实验动物110只,全部进入结果分析。①模型组10,92d胃黏膜血流均低于正常对照组(P<0.01);健胃愈疡组同期胃黏膜血流均高于模型组(P<0.01)。②健胃愈疡组和雷尼替丁组10d溃疡指数均低于模型组(P<0.01,P<0.05);模型复发组、健胃愈疡组和雷尼替丁组92d溃疡指数均高于模型组(P<0.01);健胃愈疡组10,92d溃疡指数及复发率均低于雷尼替丁组(P<0.05,P<0.01)。③模型组10,92d氨基己糖和磷脂含量均低于正常对照组(P<0.01)。健胃愈疡组10,92d氨基己糖和磷脂含量均高于模型组和雷尼替丁组(P<0.01)。溃疡指数与氨基已糖、磷脂含量呈负相关(r=-0.957,-0.960,P<0.01)。④健胃愈疡组和雷尼替丁组10d乳癌相关肽mRNA表达较正常组和假手术组提高,血小板活化因子mRNA的表达下调(P<0.01),健胃愈疡组两指标表达变化较雷尼替丁组显著(P<0.01);模型复发组、健胃愈疡组和雷尼替丁组92d乳癌相关肽mRNA、血小板活化因子mRNA的表达同组10d比较差异无显著性意义(P>0.05);模型组乳癌相关肽mRNA、血小板活化因子mRNA的表达同组10d比较差异有显著性意义(P<0.01)。结论:健胃愈疡颗粒可提高乳癌相关肽mRNA及下调血小板活化因子mRNA的表达,影响胃黏膜氨基己糖及磷脂含量,可能是其促进溃疡愈合的机制之一。     

织构复合水凝胶体系的假说

生物软组织可视为具有多层次结构的织构复合水凝胶体系(TCHS)、以水凝胶复合元件(HCE)为基本的构件(CP),通过一定的组合、排列方式构筑一系列多层次结构的不同软组织。软组织中任何层次的结构单元既可视为织构复合水凝胶体系又可视为构件。任何层次的TCHS中,构件的结构及其组合排列方式决定着该层次单元的功能。以织构复合水凝胶体系的观点考察了真核细胞、角膜和骨骼肌的多层次结构。双层网络水凝胶、皮芯复合水凝胶纤维人工肌肉模型、时空匹配可降解细胞支架等研究成果初步地证明了提出织构复合水凝胶体系观点的合理性。     

场电子发射器件中实际电子发射点密度达到实用程度的标准

目前薄膜场致电子发射平板显示方面的研究越来越深入,其制备的器件场发射性能也越来越接近实用的程度。但有关其发射点密度达到何值时,即可达到最基本可以应用的程度至今也没有一个可以直接参考的实用标准。因此提出一个这样的客观标准对于场发射显示器今后的应用来讲具有重要的实际意义。作者结合自身10多年从事碳基薄膜场致电子发射研究的经历和相应的光学理论,提出一个场电子发射器件中实际电子发射点密度可达到实用程度的标准。当薄膜场电子发射点密度在场发射器件全部发射区域内都超过106sites/cm2时,肉眼将无法在屏幕上分辨出各个发射点,此时电子发射点已连成肉眼无法分辨出的面,也就是说这时单纯从场发射均匀性上来讲,场发射器件已达到可以应用的程度。     

Practical guidelines for process validation and process control of white cell-reduced blood components: report of the Biomedical Excellence for Safer Transfusion (BEST) Working Party of the International Society of Blood Transfusion (ISBT)

Background: The increased use of white (WBC)-reduced blood components has prompted many institutions to develop quality assurance programs directed to such component preparation processes. For consistent preparation of WBC-reduced blood components that meet clinical needs as well as national standards, a program of process validation and control should be instituted. This involves controlling key factors that affect WBC reduction as well as periodic monitoring of the residual cellular content of components. Practical guidelines for the implementation of such a program are provided. Study Design and Methods: A program involving three phases of monitoring was developed by individuals belonging to an international working party of the International Society of Blood Transfusion. Results: The first phase, process validation, evaluates a minimum of 20 consecutive units (a minimum of 60 units when nonparametric measurements are used) to document the successful local implementation of a new or substantially modified process. Ongoing process control employing Levey-Jennings type control charts is used to demonstrate that the process remains stable over time. Process capability assessment and conformance with standards are evaluated once residual WBCs are counted in a sufficient number of units. This enables a facility to claim with a specified degree of confidence that a stated proportion of WBC-reduced units will meet national standards. Two approaches to determine the number of units that should be selected for counting are presented. The first approach considers units as either acceptable or not acceptable and assumes that the distribution of failed (or nonconforming) units approximates the Poisson distribution. The second approach takes into consideration the observed WBC content of the tested units, with the assumption that the residual WBC content in WBC-reduced components follows a lognormal distribution. A method to assess the lognormal distribution of residual WBCs is presented. Specific tables based on each of these approaches are provided to guide the reader in the design of a program that will verify conformance with any national standard at specific confidence levels. The approach can be generalized to other process control applications. Conclusion: Guidelines are presented for process validation, process control, and assessment of conformance in the production of WBC-reduced blood components. Policy makers retain the responsibility to establish, on the basis of the expected use of WBC- reduced components, requirements for the frequency of testing and for the proportion of prepared units that are expected with a stated degree of confidence to meet the standards. Facilities preparing WBC-reduced components can monitor key factors that influence the preparation of WBC-reduced blood, can periodically assess their conformance with the standards, and can intervene to correct adverse changes in the process. This approach can be used to ensure the consistent quality of WBC- reduced blood components.     

Microdroplet fluorochromatic assay for the enumeration of white cells (WBCs) in WBC-reduced blood components: validation and application for evaluating newly developed WBC-reduction filters

BACKGROUND: Sensitive and accurate counting methods are required to assess the residual white cells (WBCs) in WBC-reduced blood components. The Nageotte hemocytometer, widely used for this purpose, is cumbersome, and its efficacy is dependent upon the skill of the operator. The performance of a simple fluorochromatic assay using tissue-typing microdroplet trays is presented here. STUDY DESIGN AND METHODS: Undiluted samples of blood components were mixed with a fluorochromatic dye in trays. WBCs were counted under an epifluorescence microscope. The accuracy and sensitivity of this method were compared with those of the reference Nageotte hemocytometer method by using serial dilution of samples of platelets and red cells containing known concentrations of WBCs and by calculating the standard curves. The Nageotte hemocytometer and the microdroplet fluorochromatic assay (MFA) were also compared in terms of count correlation and reproducibility in 320 paired counts of plateletpheresis samples. MFA was used to evaluate a newly developed WBC-reduction red cell filter. RESULTS: The MFA for platelets and red cells was linear to 0.1 and 0.03 WBCs per microL, respectively. The linear regression line of log10 MFA versus log10 Nageotte method had a slope of 0.963, intercept of -0.04, and r2 of 0.968. The Nageotte method gave an estimation of WBC content 12 to 20 percent greater than that of the MFA. The MFA, with a larger neat sample volume, showed precision comparable to that of the Nageotte method. The filters demonstrated a median WBC reduction of 4.8 log10. CONCLUSION: The MFA is a sensitive and accurate method for quality control processes to assess the residual WBCs in WBC-reduced blood components.     

Sampling errors and the precision associated with counting very low numbers of white cells in blood components

Attention to the accurate and precise measurement of the white cell (WBC) content of transfused products has risen in response to awareness of the potential benefits of WBC-depleted components and the development of technical capabilities to produce these components. The techniques thus far reported have focused on the reliability of detecting a WBC, provided it is present in the test system. The likelihood of selecting a WBC from the product of interest for counting in the analytical system--that is, the sampling error--must also be considered. The occurrence of a WBC in a WBC-depleted component is a rare event and may be modeled with the binomial or the Poisson distribution. Several assay techniques were analyzed by using these distribution models to determine the confidence intervals of the WBC content. The 95-percent confidence intervals spanned more than 2 logs10 for some methods at 3 x 10(5) WBCs per product. It is concluded that the reporting of WBC content for research provides not only the estimate of the mean but also a confidence interval for this estimate. Quality control procedures should be designed to verify that the WBC content is less than the targeted amount and should provide an associated statement of confidence.     

血管内皮生长因子与白细胞介素12在急性白血病不同时期血清中的变化

目的:由于造血干/祖细胞发生基因突变,子代细胞增殖失控等导致的恶性血液病。血管内皮生长因子和白细胞介素12参与这一发生发展过程,检测不同时期其在急性白血病患者静脉血中的水平,有利于认知与血管新生及体液免疫的相关性。方法:随机选择2005-06/2006-04在吉林市中心医院住院的急性白血病患者25例,均经FAB分型或免疫学分型确诊,患者知情同意,并经医院伦理委员会批准。将患者分为:①初诊未治组10例。②复发组5例。③完全缓解组10例。并设9名健康查体者为正常对照。应用定量酶联免疫吸附实验测定受试者血清中血管内皮生长因子和白细胞介素12的水平,在评定白细胞介素12水平时,将初诊未治组与复发组合并为初诊复发组:①两组的发病机制相似。②两组病例数较少,单独观察没有统计学意义。结果:25例患者和9名健康对照者均进入结果分析。①初诊未治组血管内皮生长因子含量高于完全缓解组及正常对照组(P均<0.05)。②正常对照组白细胞介素12水平与初诊复发组、完全缓解组之间差异均具有显著性意义(P均<0.05)。③正常对照组血管内皮生长因子含量与白细胞介素12之间存在负相关(r=-0.9644P<0.05)。④初诊复发组、完全缓解组血管内皮生长因子和白细胞介素12含量之间均无相关性(r=-0.0883,-0.3593,P均>0.05)。结论:急性白血病患者血清中血管内皮生长因子和白细胞介素12含量与临床病情变化有关,可以作为诊断和预测急性白血病发生和复发的指标。     

A study of confidential unit exclusion

The effectiveness of the confidential unit exclusion (CUE) procedure recommended by the Food and Drug Administration has been questioned by the blood banking community. The purpose of this study was to determine whether donors were informing the blood center correctly regarding the disposition (transfuse or do not transfuse) of their donated blood. A letter explaining the confidential study and requesting permission to send the participant a questionnaire noting his or her self-exclusion choice was mailed to 230 donors who had chosen transfuse and 276 donors who had chosen do not transfuse. After consent was obtained, participants were sent a second packet and asked to indicate whether they had chosen correctly and, if not, to identify reasons for that incorrect choice. A seven-word terminology quiz made up of words from the CUE form was also enclosed. All participants who had chosen transfuse indicated that this was the correct choice. Approximately 50 percent of those who had chosen do not transfuse indicated that this was an incorrect choice; the most common reason was that "I was not paying attention." The most frequently misunderstood term was "confidential." Donors who chose do not transfuse had a significantly higher rate of error on the terminology quiz (p less than 0.01) than did those who chose transfuse.     

Oral cyclophosphamide versus chlorambucil in the treatment of patients with membranous nephropathy and renal insufficiency

We treated patients with idiopathic membranous nephropathy (iMGN) and renal insufficiency, using: (i) (n = 15) monthly cycles of steroids (1 g methyl-prednisolone i.v. on three consecutive days, followed by oral prednisone 0.5 mg/kg/day months 1, 3 and 5) and chlorambucil (0.15 mg/kg/day months 2, 4 and 6); or (ii) (n = 17) oral cyclophosphamide (1.5-2.0 mg/kg/day for 1 year) and steroids in a comparable dose. The groups were comparable in age, renal function and levels of proteinuria. During the 6 months preceding treatment, serum creatinine levels increased from 148 +/- 50 to 219 +/- 73 mumol/l in the chlorambucil group and from 164 +/- 86 to 274 +/- 126 mumol/l in the cyclophosphamide group. Median (range) follow-ups were: chlorambucil 38 months (8-71); cyclophosphamide 26 months (5-68) (NS). Renal function improved in both groups, but the improvement was short-lived in the chlorambucil group; 12 months after starting treatment, mean serum creatinine was 6.3 mumol/l lower in the chlorambucil group and 121 mumol/l lower in the cyclophosphamide group (p < 0.01). Four chlorambucil-treated patients developed ESRD, and five needed a second course of therapy, whereas only one cyclophosphamide-treated patient developed ESRD (p < 0.05). Remissions of proteinuria occurred more frequently after cyclophosphamide treatment (15/17 vs. 5/15; p < 0.01). Side-effects necessitated interruption of treatment in six patients on cyclophosphamide and in 11 on chlorambucil (p < 0.05). In our patients, oral cyclophosphamide was better tolerated than oral chlorambucil. The suggested greater efficacy of the oral cyclophosphamide regimen needs to be ascertained by longer follow-up.      

成纤维细胞生长因子Ⅱ型受体功能获得型点突变影响小鼠颅底软骨发育

目的:对比野生型小鼠和成纤维细胞生长因子Ⅱ型受体(fibroblast growth factor receptor2,fgfr2)基因S252W突变型小鼠颅底软骨的生长发育情况,探讨fgfr2功能持续增强对小鼠颅底软骨发育的影响。方法:实验于2004-03/2005-04在解放军第三军医大学大坪医院野战外科研究所创伤中心实验室,创伤、烧伤与复合伤国家重点实验室完成。①fgfr2 小鼠和C57BL/6J小鼠交配后取F1代,利用聚合酶链反应方法进行fgfr2突变型小鼠基因型的鉴定,得到野生型和fgfr2受体功能获得型点突变型两组小鼠。②标本取材及处理:分别在两组实验动物中,取胚胎期16d(E16)、胚胎期17d(E17)、新生2d(P2)、新生8d(P8)的小鼠的颅底作为标本。一部分体积分数为0.75的乙醇溶液固定。另一部分标本行40g/L多聚甲醛固定,150g/L乙二胺四乙酸溶液脱钙,常规石蜡包埋、切片。③阿尔辛-茜素红骨架染色显示颅底整体骨骼的发育情况。软骨组织染为蓝色,骨组织染为红色。④脱钙骨切片后进行苏木精-伊红染色。未脱钙骨切片后进行Calcein染色,荧光显微镜下观察钙化组织的情况。⑤使用地高辛RNA标记试剂盒按照产品说明制备Col 10(Collagen 10)地高辛标记的互补RNA探针,对颅底软骨联结进行检测软骨分化标志基因表达情况的原位杂交。⑥组织切片染色结果使用SPOT Advanced软件进行数码拍摄。结果:①实验小鼠颅底发育的大体情况:fgfr2受体功能获得型点突变小鼠从胚胎后期开始就表现出短颅现象。②阿尔辛蓝-茜素红骨架染色、Calcein染色和苏木精-伊红染色显示,fgfr2受体功能获得型点突变小鼠出生后颅底软骨联结没有出现提前融合的现象,但是颅底软骨生长发育差,软骨细胞增生区、肥大区缩短。突变小鼠的颅底软骨发育障碍导致颅底骨组织的生长发育也出现障碍。③原位杂交:Col10在野生型和突变型小鼠颅底软骨的表达部位未见明显差异,但是这个软骨细胞分化标志基因的表达强度降低。结论:fgfr2 S252W点突变能够引起小鼠颅底软骨联结处的软骨细胞发育异常,导致颅底骨骼发育畸形。fgfr2可能具有调控软骨细胞系发育的功能,从而可能在骨骼发育、骨折修复等方面发挥重要作用。