Comparison of assays for determination of peptide content for lyophilized thymalfasin*

Abstract: Precise determination of the peptide content in drug substance samples depends highly upon the particular peptide compound and methodology used. Four independent methods were evaluated and compared to determine which would produce the best experimental precision for analysis of thymalfasin (thymosin α‐1). Four different methods were evaluated including elemental analysis (CHN), quantitative amino acid analysis (AAA), high‐performance liquid chromatography (HPLC), and Kjeldahl. This study demonstrates that the AAA method is highly variable in one laboratory while quite precise in another laboratory. Similarly, HPLC results depended on the laboratory conducting the study with more precise values obtained under cGMP. On the contrary, the CHN method yielded highly precise [i.e. <2% coefficient of variation (CV)] values. As precise knowledge of protein content is fundamental for the compounding of final pharmaceutical product of a specific potency, the CHN analysis is recommended for peptide content determination of the drug substance thymalfasin.     

Diabetes mellitus as a contributory factor in oral candidosis.

It has been reported that poor glycaemic control predisposes to oral candidal infection in diabetic patients. For instance, the carriage of Candida species and the density of candidal growth in the oral cavity is frequently claimed to be increased in patients with diabetes mellitus. However, the validity of these observations remains controversial. Hence, we review and discuss here the clinical data in the literature on the relationship between diabetes and oral candidal carriage and infection, and possible mechanisms associated with its pathogenicity.     

Intravenous cyclophosphamide is the drug of choice for steroid dependent nephrotic syndrome

BACKGROUND: Steroid dependency is a major problem seen after therapy for idiopathic nephrotic syndrome in childhood. Although there is consensus about the usage of cyclophosphamide (CYC) in frequent relapsers, there is still a controversy concerning its usage in steroid-dependent nephrotic syndrome (SDNS). METHODS: In the present study, nineteen children with SDNS were treated with CYC: ten via the intravenous (i.v.) route, and nine via the oral route. Remission was then maintained with prednisolone. Oral CYC therapy consisted of CYC at a dose of 2 mg/kg per day for 12 weeks. Intravenous (i.v.) CYC therapy consisted of CYC 500 mg/m2 per month (with intravenous 3500 cc/m2 per 24 h one-third saline hydration) for 6 months. RESULTS: The cumulative dose of CYC was 168 mg/kg in the oral group and 132 mg/kg in the IV group. Daily oral CYC dose was 1.96~0.31 mg/kg, whereas i.v. CYC dose was 0.73~0.03 mg/kg. Long-term complications and side-effects such as alopecia, infection and hemorrhagic cystitis were not observed in the i.v. CYC treated group. In the long term, the dosage of prednisolone that held remission after CYC, the annualized relapse rates and the subsequent relapse time were significantly better in the i.v. CYC group, and the number of patients in remission for 2 years was significantly higher in the i.v. treated group (P<0.05). CONCLUSIONS: In SDNS, i.v. CYC has a long lasting effect with lower annualized relapse rates and longer subsequent relapse time with a lower steroid dosage required to maintain remission than oral CYC. The results of the present study showed the safety of the i.v. route, and it is the preferable treatment in noncompliant patients for its long lasting remission and simple and inexpensive follow up.     

高氧液对心肺脑复苏的作用

目的探讨高氧液静脉给药对肺心病患者心肺脑复苏成功率及预后的作用;方法采用高氧液静脉给药对26例心跳呼吸停止进行心肺脑复苏治疗的患者(观察组),与既往无高氧液静脉给药的35例心跳呼吸停止进行心肺脑复苏治疗的患者(对照组)进行复苏成功率及预后比较;结果观察组Ⅲ级和Ⅳ级复苏成功后生存11例,正常生活自理5例;对照组Ⅲ级和Ⅳ级复苏成功后生存4例,正常生活自理2例。两组Ⅲ级和Ⅳ级复苏成功率比较,观察组成功率较对照组高(P<0.01)。结论高氧液能提高心跳呼吸停止患者的心肺脑复苏成功率,可提高Ⅲ、Ⅳ级复苏的生存率。     

NIKKISO DBB-27型血液透析机液路系统工作原理及故障报警分析

分析了NIKKISO DBB-27型血液透析机液路系统的系统结构及各组成部分的工作原理,并对相关故障报警的诊断和排除进行了详细论述,为及时、准确地诊断和排除血液透析机的故障提供了理论基础。     

柔肝消瘢饮对肝硬化大鼠血清和肝组织IL-1β和IL-6含量的影响

目的：观察柔肝消瘕饮对肝硬化大鼠肝组织病理形态学和血清、肝组织IL-1β、IL-6含量的影响。方法：采用复合因素造模法复制肝硬化大鼠模型，以复方鳖甲软肝片为对照，观察柔肝消瘢饮对肝硬化大鼠肝组织病理形态学、血清和肝组织IL-1β、IL-6含量的影响。结果：与正常组比较，模型组大鼠出现肝小叶损害，纤维组织增生，假小叶形成，血清IL-1β、IL-6含量和肝组织IL-6含量均明显升高（P〈0．01或P〈0．05）；与模型组比较，各治疗组肝组织病理损害减轻，血清IL-1β和肝组织IL-6含量均有明显下降，差异有显著性意义（P〈0．01或P〈0．05）；与对照组比较，高剂量组血清IL-6含量明显降低（P〈0．05）。结论：柔肝消瘢饮能显著下调肝硬化大鼠血清IL-1β、IL-6含量和肝组织IL-6含量，具有减轻肝硬化炎症损害，改善肝组织病理形态学作用。     

HPLC法测定滑膜炎颗粒中原儿茶醛的含量

目的建立滑膜炎颗粒剂中原儿茶醛的含量测定方法质量。方法采用高效液相色谱法对方中原儿茶醛进行含量测定。结果高效液相色谱法测定结果表明原儿茶醛在0．1005～1．005μg范围内呈线性关系，平均加样回收率为99．22％；RSD为0．72％。结论本法简便，重现性好、结果可靠，可作为控制滑膜炎颗粒剂的质量方法。     

丹参营养器官中脂溶性成分的动态变化规律

目的 测定丹参营养器官中丹参酮Ⅰ和丹参酮Ⅱ_A的含量,并揭示丹参生长过程中,丹参酮Ⅰ和丹参酮Ⅱ_A的动态变化规律.方法 采用反相高效液相色谱法,以Zorbax SB-C_(18)(150mm×4.6mm, 5.0μm)为色谱柱;甲醇-0.4%磷酸为流动相,梯度洗脱,流速为1.0 ml/min;二极管阵列检测器:检测波长分别为246, 270 nm;柱温为30℃.结果 丹参叶和茎中均未检出丹参酮Ⅰ和丹参酮Ⅱ_A,根中丹参酮Ⅱ_A和丹参酮Ⅰ的含量变化呈"单峰"曲线,丹参酮Ⅱ_A的含量在7月份最高,为2.94%,丹参酮Ⅰ含量在5月份最高,为0.78%.结论 4月至7月是丹参根中丹参酮积累的关键时期.     

Chronic GVHD in minor salivary glands and oral mucosa: histopathological and immunohistochemical evaluation of 25 patients

BACKGROUND: Graft-vs.-host disease (GVHD) is the major cause of morbidity and mortality in patients undergoing allogeneic Bone Marrow Transplantation (BMT). The aim of our study was to identify the most relevant histological features for diagnosis of chronic Graft-vs.-Host Disease (cGVHD) in oral mucosa and minor salivary glands of 25 patients, as well as to evaluate the immunophenotype of the inflammatory cells. METHODS: Sixteen patients that were submitted to allogeneic BMT but did not present cGVHD were selected as a control group. The sections were studied on H & E and CD68, CD45, CD4, CD8, CD20 staining. RESULTS: The most frequent histologic findings in oral mucosa at the day of diagnosis of cGVHD were: hydropic degeneration of the basal layer of the epithelium, apoptotic bodies, lymphocytic infiltration, and focal or total cleavage between the epithelial and connective tissue. In the labial salivary glands (LSG), lymphocytic infiltration, acinar loss and fibrosis were the main alterations. Cytotoxic CD8-T cells and macrophages were predominant both in the epithelium and connective tissue, as well as in minor salivary glands. CONCLUSIONS: Histological features were useful in the diagnosis of oral cGVHD. It is suggested that CD8-T cells and macrophages play important role in the pathogenesis of the disease.     

Ploidy analysis of field cancerization and cancer development in the hamster cheek pouch carcinogenesis model

BACKGROUND: The hamster cheek-pouch carcinogenesis model is a well-known animal system that closely mimics the development of premalignant and malignant lesions in human oral cancer. Our aim was to numerically characterize the premalignant and malignant lesions and expressions of field cancerization in this model using ploidy as the end-point. METHODS: To study the DNA content and proliferation status of the cells in this model we assessed the Feulgen reaction and the immunohistochemical reaction for 5-bromo-2-deoxiuridine (BrdU) in different histological areas of serial tissue sections of the cheek pouches of animals injected with BrdU. RESULTS: Ploidy values were higher in cancerized epithelia with no unusual microscopic features (NUMF), in preneoplastic and tumor areas than in control epithelia. The aneuploidy index was higher in NUMF areas than in control and differed significantly from control in preneoplastic areas and carcinoma. CONCLUSIONS: The unexpected alteration in DNA content observed in NUMF epithelia is of great relevance as a biomarker of field cancerized areas.