Analysis of gene expression in human bullous keratopathy corneas containing limiting amounts of RNA

PURPOSE: To validate the use of polymerase chain reaction (PCR)-amplified full-length cDNA as a substitute for mRNA in nucleic acid array and gene expression analysis. METHODS: Total RNA was isolated from age-matched normal autopsy corneas and pseudophakic bullous keratopathy (PBK) corneas. Full-length cDNA was generated and PCR amplified using the Smart cDNA synthesis technology. Southern blot analysis of this cDNA was compared with Northern blot analysis of the RNA. Amplified cDNA was used to probe a commercial gene array. By immunohistochemistry, the expression pattern of several adhesion molecules represented on the array was assessed. RESULTS: The cDNA produced by the Smart cDNA system gave results very similar to those of northern blot analysis when examined for beta2-microglobulin, Rab geranylgeranyl transferase, and tenascin-C. This cDNA obtained from normal or PBK corneas was labeled and used to probe a 588 gene array (Clontech). Among other differences, beta6 integrin was detected only with the PBK probe, beta-catenin was markedly elevated in PBK, and beta4 integrin appeared to be reduced in PBK. Immunohistochemical patterns of these proteins were consistent with the hybridization signals on the gene array. CONCLUSIONS: Smart cDNA synthesis and nucleic acid arrays were combined and validated for the first time to identify differential gene expression in normal and diseased corneas. These techniques require very little RNA such as that equivalent to a half of a single cornea, which is useful when the amount of tissue is limiting. Altered expression of adhesive proteins beta6 integrin and beta-catenin may be related to the formation of epithelial bullae and microcystic changes in PBK patients.     

Selective toxicity of the tricyclic thiophene NSC 652287 in renal carcinoma cell lines: differential accumulation and metabolism

The tricyclic compound 2,5-bis(5-hydroxymethyl-2-thienyl)furan (NSC 652287) has shown a highly selective pattern of differential cytotoxic activity in the tumor cell lines comprising the National Cancer Institute (NCI) Anticancer Drug Screen. The mechanism underlying the selective cytotoxicity is unknown. We hypothesized that differential sensitivity to the compound observed in several renal tumor cell lines could be the result of selective accumulation or differential metabolism of this agent. We demonstrated here that the capacity of certain renal cell lines to accumulate and retain the compound, determined by accumulation of [14C]NSC 652287-derived radioactivity and by flow cytometric determination of unlabeled compound, paralleled the sensitivity of the renal cell lines to growth inhibition by NSC 652287: A-498 > TK-10 > ACHN approximately/= to UO-31. The ability of the cell lines to metabolize [14C]NSC 652287 to a reactive species capable of binding covalently to cellular macromolecules also directly correlated with sensitivity to the compound. Different patterns of metabolites were generated by relatively more drug-sensitive cell lines in comparison with drug-resistant cell lines. The metabolizing capacity for NSC 652287 was localized primarily to the cytosolic (S100) fraction. The rate of metabolism in the cytosolic fraction from the most sensitive renal cell line, A-498, was faster than that observed in the cytosolic fractions from the other, less sensitive cell lines. The data support the hypothesis that both selective cellular accumulation and the capacity to metabolize NSC 652287 to a reactive species by certain renal carcinoma cell types are the basis for the differential cytotoxicity of this compound class.     

Anomalies of centriolar derivatives manifest in spermatic flagella and respiratory cilia of the stallion.

Anomalies of centriolar derivatives were identified in ejaculates and testicular and tracheal biopsies of a sterile stallion, using light, scanning, and transmission electron microscopy. LM revealed that over half the sperm population had only a vestigial or no tail, while the rest had tails of variable length and shape. The vestigial tail was represented by its anlage, which was implanted on the nucleus and differentiated up to capitulum and collum stage. The stunted tail had an axoneme and its derivatives, but was short in all tail segments. Regardless of the tail length or shape, virtually all axonemes were devoid of the central tubular complex ("9 + 0" defect). Abnormal tail segmentation was associated with missing or defective flagellar sheaths and a profusion of extraneous dense fibers, which contributed to the knobby, bulbous, or lobuliform tail configurations. The gradient of flagellar anomalies seems associated with the inability of the distal centriole to implant on the plasmalemma, to produce the axoneme, or maintain its growth, and to induce the normal differentiation of periaxonemal structures. In contrast to sperm, the tracheal epithelium displayed moderate changes, which are manifest in circumscribed rarefaction of cilia, increased incidence of compound cilia, and disturbed orientation of cilia regarding the plane of central tubular complex. The tracheal cilia were free of "9 + 0" defect.     

蒙药忠论-5胶囊总生物碱测定及其体外溶出度研究

目的 考察蒙药忠伦-5胶囊中总生物碱含量及其体外溶出情况。方法 以苦参碱为对照品，利用酸性染料比色法测定总生物碱；以水为介质，用转篮法考察其体外溶出度。结果 含量测定方法精密度为RSD＝1．89％，平均回收率为100．89％，RSD＝3．49％。；苦参总生物碱相对累积溶出度与溶出时间相关显著(P＜0．01)，T50＝24min，Td＝31min，m＝1．32。结论 用上法测定总生物碱含量及体外溶出度，对评估忠伦-5胶囊质量有实用价值。     

A prospective trial of short‐fractionation radiotherapy for the palliation of liver metastases

The purpose of this study was to prospectively examine the effectiveness and tolerability of a simple radiotherapy technique for the palliation of symptomatic liver metastases. Twenty‐eight patients with symptomatic liver metastases were enrolled from seven centres, and received targeted (partial or whole) liver irradiation consisting of 10 Gy in two fractions over 2 days. Symptoms at baseline were hepatic pain (27 patients), abdominal distension (19), night sweats (12), nausea (18) and vomiting (eight). Twenty‐two patients (76%) had failed previous treatment with chemotherapy, hormonal therapy and/or high‐dose steroids. Symptoms and potential toxicities were prospectively assessed at the time of treatment, then 2, 6 and 10 weeks later. Individual symptom response rates were 53?66% at 2 weeks. Partial or complete global symptomatic responses were noted in 15 patients (54%) overall. The treatment was well tolerated with two patients (7%) experiencing grade 3 toxicity (one vomiting and one diarrhoea); however, four patients reported temporary worsening of pain shortly after treatment. This simple and well‐tolerated treatment achieves useful palliation.     

Aberrant crypt focus promotion and glucose intolerance: correlation in the rat across diets differing in fat, n-3 fatty acids and energy

McKeown-Eyssen (Cancer Epidemiol. Biomarkers Prevent., 3, 687-695, 1994) and Giovannucci (Cancer Causes Control, 6, 164-179, 1995), noting the striking similarity in lifestyle risk factors for colorectal cancer and insulin resistance, proposed that the hyperinsulinemia, glycemia and hypertriglyceridemia associated with insulin resistance promotes colon cancer. To compare the effect of diet on colon cancer promotion and insulin resistance in the F344 rat, we assessed the effect of fat, n-3 fatty acids and energy in pairwise comparisons on average size of aberrant crypt foci (ACF) and on glucose intolerance in the same animals in a single experiment. Diets high in fat and energy increased and diets with increased n-3 fatty acids and calorie restriction decreased both ACF growth and glucose intolerance compared with control diets. The measures of promotion of colon cancer and insulin resistance were strongly correlated (n = 98, r = 0.67, P < 0.001). In addition, both were highly correlated with daily energy intake (r = 0.62 and 0.66) and were also correlated with basal (post-prandial) insulin, glucose and triglycerides (r = 0.31-0.53, P < 0.01). We concluded that ACF growth and glucose intolerance are correlated for a wide range of diets and that increased circulating energy (glucose and triglycerides) may lead to both colon cancer promotion and insulin resistance.      

多非替利衍生物CPU 228(V03)对大鼠心室肌细胞钙瞬变的调节

目的在β-受体激动情况下,研究多非替利衍生物CPU 228对电刺激触发心肌细胞的胞浆Ca2+浓度([Ca2+]i)变化及钙瞬变动力学参数的影响.方法游离单个大鼠心室肌细胞,Fluo-3/AM负载,电刺激诱发心室肌细胞钙瞬变,定量测定心室肌细胞内Ca2+浓度变化,异丙肾上腺素(isoproterenol, ISO)100nmol·L-1激动β-受体,观察CPU 228 1 μmol·L-1对钙瞬变动力学参数、[Ca2+]i及钙负荷水平的影响.结果CPU 228 1 μmol·L-1对大鼠心室肌细胞[Ca2+]i的影响不明显(P>0.05);ISO 100 nmol·L-1显著升高大鼠心室肌细胞[Ca2+]i和细胞内钙负荷水平,缩短钙瞬变时程,并且增加钙瞬变的消除速率.CPU 228 1 μmol·L-1显著抑制ISO的上述作用.结论在β-受体激动情况下,CPU 228可以降低心肌细胞[Ca2+]i,使ISO改变的钙瞬变动力学参数恢复到正常水平.     

Safety and efficacy of mangafodipir trisodium (MnDPDP) injection for hepatic MRI in adults: results of the U.S. multicenter phase III clinical trials (safety)

The short-term safety of mangafodipir trisodium (MnDPDP) injection was studied in 546 adults with known or suspected focal liver lesions. An initial contrast-enhanced computed tomography examination was followed by unenhanced magnetic resonance imaging (MRI), injection of MnDPDP (5 micromol/kg), and enhanced MRI. Adverse events were reported for 23% of the patients; most were mild to moderate in intensity, did not require treatment, and were not drug related. The most commonly reported adverse events were nausea (7%) and headache (4%). The incidence of serious adverse events was low (nine events in six patients) and not drug related. Injection-associated discomfort was reported for 69% of the patients, and the most commonly reported discomforts included heat (49%) and flushing (33%). Changes in laboratory values and vital signs were generally transient, were not clinically significant, and did not require treatment. There were no clinically significant short-term risks from exposure to MnDPDP.     

乌贼墨对小鼠脾细胞和巨噬细胞NO生成及 IFN-γ分泌水平的影响

目的：探讨乌贼墨对脾细胞和巨噬细胞NO生成及IFN-γ分泌的影响。方法：用Griess法和ELISA法分别检测了经乌贼墨灌胃处理后小鼠脾细胞和腹腔巨噬细胞培养上清中NO和IFN-γ的水平的变化。结果：乌贼墨灌胃处理小鼠后的第4、6天脾细胞及腹腔巨噬细胞(Mφ)可产生较高水平的NO；脾细胞可产生较高水平的IFN-γ，并且小鼠脾细胞NO的产生水平与IFN-γ产生水平呈正相关(r=0．98)；用LPS和L-NMMA(NO抑制剂)分别同乌贼墨灌胃第6d小鼠的脾细胞一起培养，结果显示LPS可协同IFN-γ促NO水平升高。L-NMMA可抑制小鼠脾细胞的NO生成。结论：乌贼墨可促进巨噬细胞NO生成；脾细胞NO的产生与IFN-γ分泌水平呈正相关。     

食品级番茄红素对小鼠的免疫调节功能

目的研究番茄红素对免疫系统的调节作用。方法实验小鼠随机分成3组：对照组、低剂量组（10mg／kg）、高剂量组（20mg／kg）。低剂量组、高剂量组分别用色拉油稀释的番茄红素灌胃，对照组用色拉油灌胃。30天后检测小鼠的脾淋巴细胞增殖程度、NK细胞的杀伤活性、迟发性变态反应的程度、巨噬细胞吞噬功能。结果与对照组比较，高剂量组能显著增强脾淋巴细胞的增殖能力，提高自然杀伤细胞的活性和巨噬细胞吞噬功能，增强小鼠的足跖肿胀度（P〈0．05）。除增殖反应外，低剂量组其他指标与对照组比较，差异均无显著性。结论番茄红素具有增强小鼠免疫调节的功能。