Effect of Glass Transition Temperature on the Stability of Lyophilized Formulations Containing a Chimeric Therapeutic Monoclonal Antibody

Purpose. The purpose of this study is to highlight the importance of knowing the glass transition temperature, T_g, of a lyophilized amorphous solid composed primarily of a sugar and a protein in the interpretation of accelerated stability data. Methods. Glass transition temperatures were measured using DSC and dielectric relaxation spectroscopy. Aggregation of protein in the solid state was monitored using size-exclusion chromatography. Results. Sucrose formulation (T_g ~ 59°C) when stored at 60°C was found to undergo significant aggregation, while the trehalose formulation (T_g ~ 80°C) was stable at 60°C. The instability observed with sucrose formulation at 60°C can be attributed to its T_g (~59°C) being close to the testing temperature. Increase in the protein/sugar ratio was found to increase the T_gs of the formulations containing sucrose or trehalose, but to different degrees. Conclusions. Since the formulations exist in glassy state during their shelf-life, accelerated stability data generated in the glassy state (40°C) is perhaps a better predictor of the relative stability of formulations than the data generated at a higher temperature (60°C) where one formulation is in the glassy state while the other is near or above its T_g.     

糖尿病患者微血管病变与血小板功能

测定21例有微血管病变和27倒无并发症的糖尿病人血小板功能，并与40例正常人作比较。结果发现糖尿病人，尤其是微血管病变者血浆血栓烷素（TXB2）、第八因子相关抗原（VWF）浓度及血小板膜糖蛋白（G、P）Ib显著高于对照组。提示血小板功能异常可能对糖尿病患者做血管并发症的发生起重要作用。     

1,6-二磷酸果糖治疗婴幼儿肺炎心衰疗效观察

应用脉冲多普勒超声心动图对３０例使用１，６－二磷酸果糖的肺炎心衰患儿进行心室舒缩功能指标的观测，并以２０例常规治疗的心衰患儿及２０例正常儿为对照组，评价１，６－二磷酸果糖的疗效。结果表明，１，６－二磷酸果糖能明显改善ＰＥＰ、ＰＥＰ／ＥＴ、ＰＥＰ／Ｒ－Ｒ、ＦＶＩ和ＳＶ等心室收缩功能指标及ＰＦＶＥ、Ｅ／Ａ、ＰＦＶＡ等心室舒张功能指标，使心功能恢复正常。１，６－二磷酸果糖对肺炎心衰引起的心肌缺血、缺氧有保护作用。     

增殖细胞核抗原及nm23在胃癌中的表达及意义

目的 :探讨增殖细胞核抗原 (PCNA)及nm2 3基因在胃癌中的表达及其与生物学行为的关系。方法 :应用免疫组化SP法研究 85例胃癌手术标本中PCNA及nm2 3基因的表达。结果 :在 85例胃癌中PCNA、nm2 3的总阳性表达率分别为 5 2 9% (4 4 / 85例 )和 4 4 .7% (38/ 85例 ) ,PCNA及nm2 3在胃癌中的表达与其淋巴结转移、浸润深度、分化程度有关 (P <0 0 5 ,P <0 0 1 ) ;PCNA与分化程度无关 ,P >0 0 5。结论 :PCNA及nm2 3参与了胃癌的发生、发展、浸润及转移     

99Tcm标记人源化抗人血小板膜糖蛋白Ⅲ a单克隆抗体SZ21F(ab)2血栓栓塞显像

目的用动物血栓栓塞模型显像评估99^Tc^m标记人源化抗人血小板膜糖蛋白（GP）Ⅲa单克隆抗体（简称单抗）F（ab）2片段SZ21F（ab）2的应用价值。方法通过体外抑制二磷酸腺苷（ADP）诱导的犬血小板聚集实验，评估SZ21F（ab）2与血小板结合的亲和性和特异性。以2-亚氨基噻吩盐酸盐（IT）法修饰SZ21F（ab）2分子的亚氨基，以99^Tc^m-葡庚糖酸钠（GH）转换络合法进行标记。分别用快速薄层层析（ITLC）法和ELISA测定标记物的放化纯及生物活性。对3条肺动脉血栓和8条（包括上述3条）后肢深静脉血栓栓塞比格犬模型进行显像研究。结果SZ21F（ab）2抑制ADP诱导的犬富含血小板血浆（PRP）聚集实验的半数有效剂量（IC50）为（2．49±1．88）μg／ml。ITLC法测得标记物的放化纯为90％～95％，ELISA测定结果表明标记后抗体保留了免疫活性。注射显像剂后1h，肺动脉血栓部位和后肢深静脉血栓部位均出现放射性浓聚。注射后3h，在体显像肺动脉血栓和后肢深静脉血栓栓塞模型的放射性靶／本底（T／B）比值分别为3．03±1．18和3．51±0.62。肺动脉血栓部位和后肢深静脉血栓部位的每克组织百分注射剂量率（％ID／g）分别为0．083％ID／g和0．076％ID／g。体外检测结果表明：肺动脉血栓与正常肺组织的单位质量放射性比值平均为12．8，后肢深静脉血栓与血液的单位质量放射性比值平均为5．2，与肌肉的比值平均为127.0。结论99^Tc^m-SZ21F（ab）2与人血小板具有较高的亲和力，有潜在的血栓显像应用前景。     

喉癌患者血小板聚集功能的测定

目的 探讨喉癌与血小板聚集功能的关系。方法 采用比浊法，对50例喉癌患进行血小板聚集功能的检测并以良性肿瘤病人、喉慢性炎性疾病病人及正常人作为对照，应用SAS计算机系统进行分析。结果 与其他组病人相比，喉癌组血小板聚集功能增强并且随着临床分期的增高有增强趋势。结论 喉癌病人的血小板聚集功能有一定的增强；测定血小板聚集功能对恶性肿瘤的疗效判定、预后评估及预测转移有一定的意义；血小板抑制剂有望成为治疗肿瘤的新方法。     

小剂量阿司匹林对缺血性脑血管病患者血小板聚集功能的影响

目的 :观察小剂量阿司匹林 (ASA)对ICVD患者血小板聚集功能的影响。方法 :用花生四烯酸 (AA ,5 0 0 μmol·L-1)、二磷酸腺苷 (ADP ,5 μmol·L-1)、肾上腺素 (EPN ,5 μmol·L-1)和胶原 (COL ,2 μg·mL-1)做诱导剂检测ICVD患者服用不同剂量ASA组 ( 2 5、5 0和10 0mg·d-1)的血小板聚集率。结果 :各ASA组对AA、COL诱导聚集的变异系数较大。对AA诱导聚集无显著抑制者的比例分别为2 5mg·d-1组 44 4%、5 0mg·d-1组 2 9 6%、10 0mg·d-1组 2 9% ,其中 5 0mg·d-1组中合并糖尿病的例数较显著抑制者中多。结论 :ICVD患者中小剂量ASA作用个体差异较大 ,部分与自身危险因素有关。提示临床中ASA的效果需要实验室检测并应个体化     

A multiparametric index of platelet in vitro aggregation in cerebrovascular disease

148 patients with various forms of cerebrovascular disease (CVD) were studied by means of a multiparametric analysis ofin vitro platelet aggregation, based on the following six parameters: ADP and epinephrine primary and secondary aggregation thresholds and percent maximum aggregation induced by optimal concentrations of ADP and epinephrine. These patients were assigned to four study groups, according to clinical diagnosis supported by CT scan, of transient ischemic attack and reversible neurological deficit (TIA-RIND), or completed stroke, in the presence or absence respectively of antiplatelet medical treatment at the time of the study. A statistically significant increase of thein vitro platelet aggregation was found in 44.4% of the untreated TIA-RIND patients and in 33.9% of the untreated stroke patients. However this last group showed a higher percentage of very marked hyperaggregation. Differences between the two treated study groups and controls were not signicant. No difference was found in collagen-and ristocetin-induced aggregation between the patient groups and the controls.

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Sommario 148 pazienti con varie forme di malattia cerebrovascolare, sono stati studiati con analisi multiparametriche dell'aggregazione piastrinica in vitro sulla base dei seguenti sei parametri: le soglie di aggregazione primaria e secondaria e l'aggregazione massima percentuale indotta da ADP ed Epinefrina. Questi pazienti sono stati suddivisi in 4 gruppi di studio in accordo con la diagnosi clinica confortata dai dati della TAC e cioè: TIA, RIND, o rammollimento in presenza o in assenza rispettivamente di un trattamento antiaggregante nel momento dello studio. è stato trovato un aumento statisticamente significativo dell'aggregazione in vitro delle piastrine nel 44.4% dei casi TIA, RIND non trattati e nel 33,9% dei casi di rammollimento non trattati. Quest'ultimo gruppo, però, ha dimostrato una più alta percentuale di iperaggregazione molto marcata. Le differenze tra i 2 gruppi di studio trattati con antiaggreganti e i controlli non erano significative. Inoltre nessuna differenza è stata riscontrata tra i gruppi e i controlli nell'aggregazione indotta da collageno e ristocetina.

     

特异性血小板抗体检测的实验研究

目的　建立一种简易、特异性强的血小板抗体检测方法。方法　利用磷酸氯喹溶液对血小板进行处理 ,去掉血小板相关抗原 (HLA抗原 ) ,用只含特异性抗原的血小板包被聚乙烯板来捕获血小板抗体。结果　与常用的SEPSA和淋巴细胞毒实验 (LCT)比较 ,该方法只表现血小板特异性抗体反应。结论　本方法能够鉴别血小板相关抗原的抗体和血小板特异性抗原的抗体 ,为临床血小板减少疾病的鉴别诊断、非溶血性发热反应和血小板输注无效的原因提供有用的参考依据     

Score Statistic to Test for Genetic Correlation for Proband-Family Design

In genetic epidemiological studies informative families are often oversampled to increase the power of a study. For a proband‐family design, where relatives of probands are sampled, we derive the score statistic to test for clustering of binary and quantitative traits within families due to genetic factors. The derived score statistic is robust to ascertainment scheme. We considered correlation due to unspecified genetic effects and/or due to sharing alleles identical by descent (IBD) at observed marker locations in a candidate region. A simulation study was carried out to study the distribution of the statistic under the null hypothesis in small data‐sets. To illustrate the score statistic, data from 33 families with type 2 diabetes mellitus (DM2) were analyzed. In addition to the binary outcome DM2 we also analyzed the quantitative outcome, body mass index (BMI). For both traits familial aggregation was highly significant. For DM2, also including IBD sharing at marker D3S3681 as a cause of correlation gave an even more significant result, which suggests the presence of a trait gene linked to this marker. We conclude that for the proband‐family design the score statistic is a powerful and robust tool for detecting clustering of outcomes.