Molecular analysis of induced idiotypes associated with autoanti-thyroglobulin

Idiotype (Id) and autoanti-thyroglobulin were induced in different strains of mice by priming with anti-Id to monoclonal anti-thyroglobulins (D8 and G4) and challenged with a subimmunogenic dose of thyroglobulin (Tg). Both D8.Id and G4.Id were induced in CBA mice by priming with the appropriate anti-Id, but only priming with anti-D8.Id also induced an increase in anti-Tg. D8.Id was induced in other strains by the same schedule but it only appeared to be associated with anti-Tg in 129 and, to a lesser extent, BALB/c mice, both of which have the allotype Iga. The extent of the overlap between the D8 Id and the anti-Tg was estimated and shown to be greatest in the CBA strain from which the D8 clone was originally derived. Spectrotypic analysis of the induced Ids in CBA mice showed that some of the D8.Id, but none of the G4.Id, was identical to the original clonotype, implying that CBA mice normally have cells which can be induced to produce D8.Id-positive autoanti-Tg, which are normally weakly expressed or regulated. The observation that anti-D8.Id priming in some strains increased D8.Id-negative anti-Tg responses suggests that the D8.Id may also be associated with anti-Tg T-cells.     

A Radioimmunoassay Procedure for the Determination of the Antiviral Nucleoside DHPG (9-[(l,3-Dihydroxy-2-propoxy)-methyl]guanine) in Plasma or Serum

A procedure is described that is suitable for the radioimmunoassay (RIA) of 9-[(l,3-dihydroxy-2-propoxy)-methyl]guanine (DHPG) in plasma or serum at concentrations as low as 0.7 ng/ml (2.75 × 10^–9 M). Antiserum was prepared by coupling DHPG monohemisuccinate to bovine serum albumin and immunizing rabbits with the resulting conjugate. The antibodies did not show significant cross-reactivities with structurally related endogenous compounds. For RIA, tritium-labeled DHPG was used as the tracer and charcoal–dextran was used to separate the free and bound fractions. No purification of samples was required prior to RIA. The accuracy of the method was assessed by adding known quantities of DHPG to DHPG-free plasma and determining the ratio of measured to added analyte. Linear regression analysis for the concentration range 0.0007 to 15.0 µg/ml yielded the following equation; y = 0.90 x + 0.033 (r = 0.999). Additional validation was obtained from studies in which DHPG was administered to a monkey, mice, dogs, and rats, and plasma-clearance profiles were determined by RIA and high-performance liquid chromatography (HPLC). The results obtained by RIA were in good agreement with those obtained by HPLC.     

Monoclonal radioimmunoassay for hepatitis B surface antigen in sera of children with acute leukemia

Hepatitis B surface antigen (HBsAg) was detected by a monoclonal antibody radioimmunoassay in sera from five of 43 children (11.6%) with acute leukemia, who were negative by conventional assay. None of the nine positive sera had evidence of reactivity for HBV-DNA or DNA-polymerase activity. No correlation was found between the presence of HBsAg in serum by monoclonal RIA and the behaviour of anti-viral antibodies. Twenty-two children could be studied for liver HBsAg by immunofluorescence, and nine of them (40.9%) were positive, including three patients having HBsAg reactivity in serum. These data indicate that monoclonal antibodies increase the sensitivity of RIA for the detection of serum HBsAg in children with acute leukemia, who previously have frequently been found to have an atypical hepatitis B virus (HBV) serology.     

男性不育与性功能障碍患者生殖激素水平的改变

目的 :探讨男性不育与性功能障碍患者生殖激素水平的改变状况及其临床意义。方法 :采用体外放射免疫分析法测定患者血清生殖激素 (T、E2、LH、FSH)水平。结果 :(1)男性不育症患者血清T值与正常男性无差异 (p >0 0 5 ) ,E2 、LH和FSH水平升高 (p <0 0 0 1) ,T/LH、T/FSH降低 (p <0 0 0 1) ;T与E2 水平相关性减弱 ,FSH与LH相关性增强。 (2 )男性性功能障碍患者血清T值降低、E2 升高、T/LH降低 ,LH和FSH水平与正常男性无异 ;T与E2 水平呈负相关 ,E2 与LH由正相关变为负相关。结论 :男性不育症除了与丘脑 垂体 性腺轴功能失调有关外 ,还可能伴有睾丸间质细胞的损害 ;而E2 升高、T降低则可能在男性性功能障碍中扮演着十分重要的角色。因此生殖激素水平测定将有助于男性不育症和性功能障碍的诊断。     

基因重组人碱性成纤维细胞生长因子药物动力学实验研究──家兔静脉快推给药室分析

目的：研究家兔静脉快推基因重组人碱性成纤维细胞生长因子（ｒｈｂＦＧＦ）后血药浓度随时间的变化关系,并对其代谢动力学参数进行估计。方法：家兔静脉一次快推ｒｈｂＦＧＦ４０００ＩＵ（１０μｇ）,给药后１０,２０,３０ｍｉｎ及１．０,１．５,２．０,３．０,４．０,６．０,８．０ｈ耳缘静脉采血,放免分析法检测血中各个时间点的药物浓度。用数学方法拟合血药浓度－时间的函数关系,并估计药物动力学参数。结果：家兔静脉一次快推给药后血中ｒｈｂＦＧＦ浓度属双室模型一级动力学特征,其血药浓度－时间函数的解析表达式为：Ｃｔ＝９０．１１５６ｅ－２．０１０７ｔ＋１３．０７０７ｅ－０．１７８３ｔ。结论：本研究为指导ｒｈｂＦＧＦ的临床应用提供药物动力学的基本依据。     

Evaluation and comparison of therapeutic monitoring of whole-blood levels of cyclosporin A and its metabolites in renal transplantation by HPLC and RIA methods

Background: The aim of the work was to evaluate the possibility to estimate the level of cyclosporin A (CyA) metabolites as the difference of radioimmunoassay (RIA) non-specific and RIA specific methods. Methods: Blood samples of renal transplant patients were analyzed by three different methods: RIA specific method (CYCLO-Trac, DiaSorin, USA) (RIA_SP), RIA non-specific method (Immunotech, Czech Republic) (RIA_NS), and high performance liquid chromatography (HPLC) method. Results: Although values obtained by RIA_SP correlated well those obtained by HPLC (RIA_SP=0.995·HPLC+9.68; r²=0.962, n=448), the results of HPLC methods were lower by 8%. The values obtained by RIA_NS were 2.57 times higher than the values obtained by RIA_SP (RIA_SP=0.356RIA_NS; r²=0.713, n=448). The ratio (CyA+CyA metabolites)/(CyA) calculated as the ratio RIA_NS/RIA_SP values for 42 renal transplant patients was relatively stable for each particular patient. The sum of selected CyA metabolites (M1+M17+M21) measured by HPLC correlated well with that estimated from the difference of RIA_NS−RIA_SP: HPLC_metab=0.921·(RIA_NS−RIA_SP)+21.3; (r²=0.746, n=448). Conclusion: The combination of both the specific and non-specific methods for the determination of CyA presents an improved means for the TDM of CyA and CyA metabolites in renal transplant patients. Moreover, a combination of both methods can help to elucidate some unexpected events, such as the persistence of high cyclosporin blood levels.     

Modulation of aflatoxin toxicity and biomarkers by lycopene in F344 rats

Modulation by lycopene of aflatoxin B(1) (AFB(1))-induced toxic effects, metabolism, and metabolic activations was studied in young F344 rats. Animals were pretreated orally with either corn oil (control group) or lycopene [100 mg/kg body weight (b.w.), intervention group] 5 days/week for 2 weeks. Control animals were then treated daily with AFB(1) (250 microg/kg b.w) alone. Intervention animals were administered lycopene (100 mg/kg b.w.) at 1 h following a daily treatment with AFB(1) (250 mug/kg b.w.). Pretreatment and intervention with lycopene significantly reduced the toxic effect caused by AFB(1) and greatly modulated AFB(1) metabolism and metabolic activation. Urinary excretion of AFB(1) phase 1 metabolites, AFM(1), AFQ(1), and AFP(1), was significantly decreased in lycopene-treated animals. Formation of serum AFB(1)-albumin adducts was also significantly reduced. The rate of reduction was from approximately 30% on day 1 (p<0.05) to 67.7% on day 15 (p<0.001). Lycopene intervention also significantly reduced formation of AFB(1)-DNA adducts in liver compared to control animals, with the highest reduction (52.7%) occurring on day 3 (p<0.05). Levels of AFB(1)-N(7)-guanine excreted in urine were also significantly decreased. Urinary excretion of the phase 2 detoxification metabolite, AFB(1)-mecapturic acid, was significantly increased in lycopene-intervened animals. AFB(1)-induced urinary excretion of 8-hydroxydeoxyguanosine was also reduced to 50% on day 7 after lycopene intervention. Collectively, these results suggest that inhibition of phase 1 metabolism and metabolic activation, as well as induction of phase 2 detoxification enzyme activity are the potential mechanisms for the chemopreventive effects of lycopene.     

放射免疫法血清学指标对慢性乙肝患者肝纤维化诊断的评价

目的评价放射免疫法（RIA）检测血清层黏连蛋白（LN）、透明质酸（HA）、Ⅲ型前胶原（PCⅢ）和Ⅳ型胶原（Ⅳ-C）的诊断价值,为临床选择灵敏、特异的肝纤维化检测方法提供理论依据。方法应用放射免疫法检测119例慢性乙型肝炎患者血清中层黏连蛋白（LN）、透明质酸（HA）、Ⅲ型前胶原（PCⅢ）和Ⅳ型胶原（Ⅳ-C）的血清学含量;对肝穿刺活检标本进行病理组织学检查,包括HE染色、网状纤维染色及肝纤维化病理分期等;采用Spearman等级相关分析比较各血清指标检测结果与其相对应乙肝患者肝纤维化病理分期的关系。结果 LN、HA、PCⅢ、Ⅳ-C与肝组织纤维化病理分期的等级相关系数分别为0.073（P=0.390）、0.260（P=0.002）、0.138（P=0.104）、0.120（P=0.156）。结论血清肝纤维化放射免疫法的检测结果除HA外,LN、PCⅢ、Ⅳ-C的血清学水平与慢性乙肝患者肝纤维化的病理分期无相关性。     

Inhibitory effects of salvianolic acid B on CCl4-induced hepatic fibrosis through regulating NF-κB/IκBα signaling

Ethnopharmacological relevance

Hepatic fibrosis, a precursor of liver cirrhosis, is a consequence of severe liver damage that occurs in many patients with chronic liver diseases. Salvianolic acid B (SA-B) is one of water soluble compounds derived from Salvia miltiorrhiza Bunge (Danshen in Chinese) widely used for chronic liver diseases. In this study we investigated the protective effects of SA-B on CCl₄-induced hepatic fibrosis.

Materials and methods

Hepatic fibrosis in rats was induced by carbon tetrachloride (CCl₄). Rats were divided into four groups, including normal controls (N group), model (M group), low SA-B of 10 mg/kg body weight (L group), or high SA-B of 20 mg/kg body weight (H group). After 6 weeks, macroscopic features of the liver and weight ratio of liver to body were measured. Liver fibrosis of the rats was evaluated by HE and Massion staining. Activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL) were checked with automated biochemistry analyzer. Serum levels of hyaluronic acid (HA), type IV collagen (IV-C), Laminin (LN) and procollagen III peptide (PIIIP) were detected by radioimmunoassay (RIA). The expression of NF-κB and IκBα was detected by western blotting.

Results

SA-B was shown to reduce CCl₄-induced hepatic fibrosis in rats. The serum levels of ALT, AST, and TBIL were significantly lower in the SA-B treatment groups than in the M group. Compared the M group, the serum levels of HA, LN, IV-C and PIIIP were decreased markedly after treatment with SA-B, especially in the H group. Treatment with SA-B at 10–20 mg/kg (L and N groups, respectively) dose-dependently decreased the expression of NF-κB in the nucleolus and increased the expression levels of NF-κB and IκBα protein in the cytoplasm compared to that of the M group.

Conclusions

This study reveals that SA-B could prevent the progression of liver angiogenesis and alleviate liver fibrosis possibly by regulating the expression of NF-κB and IκBα.