我院2009～2013年口服降糖药用药分析

目的分析我院2009～2013年近5年来降糖药用药情况,为临床合理用药提供参考。方法通过医院信息管理系统,对2009～2013年我院口服降糖药的销售金额、用药频度（DDDs）、日均费用（DDDc）等进行回顾性分析。结果我院近5年来口服降糖药销售金额排序前2位的是磺酰脲类、胰岛素增敏药;罗格列酮的销售金额始终位居第1位;DDDs值排名位居第1位的是二甲双胍,而DDC排名最低;其中瑞格列奈的用药金额排序和DDDs排序同步性较好。结论我院的口服降糖药使用基本合理,新药用量呈上升趋势,质优价廉的降糖药在临床应用中占优势。     

A comparison of statistical approaches to the derivation of EC3 values from local lymph node assay dose responses.

Effective risk assessment and management of allergic contact dermatitis require three key factors: adequate hazard identification, measurement of the relative potency of identified hazards and an understanding of the nature, extent and duration of exposure. Suitable methods for hazard identification, such as the murine local lymph node assay (LLNA) and the guinea-pig maximization test, are well established and conditions of human exposure normally can be well anticipated. Thus, the need is for a robust and quantitative method for the estimation of relative skin sensitizing potency. One possible approach is via the analysis of LLNA dose-response data. In the LLNA, contact allergens are defined currently as those chemicals that cause a threefold or greater increase in lymph node cell proliferative activity compared with concurrent vehicle-treated controls. It is possible to estimate the concentration of a sensitizer required to generate a threefold stimulation of proliferation in draining lymph nodes; such a concentration is known as the EC3 value. Using a variety of statistical approaches to derive EC3 values from LLNA dose-response data for 10 chemicals, it has been demonstrated that simple linear interpolation between the values either side of the threefold stimulation index provides a robust assessment of the EC3 value without the need for recourse to more sophisticated statistical techniques. Provided that the appropriate concentrations of test chemical have been selected, EC3 values obtained in this way are reproducible both within and between laboratories and form the basis for examination of the utility of this approach for the estimation of relative skin sensitizing potency.     

Sensitivity of different cell lines to phototoxic effect of disulfonated chloroaluminium phthalocyanine.

Photodynamic therapy (PDT) is a treatment for cancer involving three key components: sensitizer, light and tissue oxygen. A sensitizer is a chemical compound that can be excited by light of a specific wavelength. Phthalocyanine ClAlPcS(2), belonging among the promising second generation of sensitizers, was evaluated as an inducer of photodamage on NIH3T3 (mouse fibroblasts), B16 (mouse melanoma), MCF7 (human breast adenocarcinoma) and G361 (human melanoma) cell lines. A semiconductor laser was used as a source for evocation of the photodynamic effect. We report the influence of various concentrations of the sensitizer in combination with laser irradiation on the photodamage of cells. Viability of cells was determined by means of molecular probes (Calcein AM and ethidium homodimer) for fluorescence microscopy. The quantitative changes of cell viability in relation to sensitizer concentrations and laser irradiation were proved by fluorometric measurement. We detected phototoxicity evoked by laser irradiated sensitizer in all studied cell lines. In addition, the viability studies showed that G361 melanoma cells and MCF7 breast adenocarcinoma cells were more sensitive than NIH3T3 mouse fibroblasts and B16 mouse melanoma to photodynamic damage induced by ClAlPcS(2).     

A potential in vitro epidermal equivalent assay to determine sensitizer potency

Most in vitro assays aim to distinguish sensitizers from non-sensitizers. Few aim to classify sensitizers according to potency. Here, we describe a potential method for classifying sensitizers according to their irritant potency with the aid of in house epidermal equivalents (EE). Sixteen sensitizers were applied topically in a dose response to EE for 24 h. The EE-EC₅₀ value (effective chemical concentration required to reduce cell viability by 50%) and the EE-IL-1α_10× value (chemical concentration which increases IL-1α secretion by 10-fold) were calculated. From 16 sensitizers, EE-EC₅₀ and/or EE-IL-1α_10× values were obtained from 12 skin sensitizers. EE-EC₅₀ and IL-1α_10× values decreased in proportion to increasing sensitizer potency. The in vitro assay correlated with existing in vivo mouse and human sensitization data (LLNA, HRIPT), and showed low intra- and inter-experimental variability. Additionally DNCB and resorcinol were correctly assessed as extreme and moderate sensitizers using commercial EE (EST1000™ and RHE™). In conclusion, our data supports the view that irritancy may in part be a factor determining sensitizer potency. Since this assay does not distinguish sensitizers from non-sensitizers, its potential application is in a tiered strategy, where Tier 1 identifies sensitizers which may then tested in Tier 2, this assay, which determines sensitizer potency.     

2008-2010年我院降糖药物应用分析

目的:了解和分析我院降糖药物的使用情况和发展趋势,为临床合理用药提供参考。方法:以我院2008-2010年降糖药物的出库记录为依据,采用金额排序法和频度分析法,分析统计降糖药物的年消耗金额,各类药物消耗金额排序以及DDDs(用药频度)、DDC(限定日费用)排序。结果:3年内降糖药物销售金额逐年上升,但降糖药物销售金额占药品总销售金额的比例平衡;胰岛素类的销售金额排在第一位,其中诺和灵30R笔芯销售金额连续3年位居首位;磺酰脲类格列美脲的DDDs连续2年位居首位;胰岛素增敏剂吡格列酮(卡司平)的销售金额排序和DDDs逐年上升。结论:我院临床使用的降糖药以胰岛素类、α-糖苷酶抑制剂、磺酰脲类、胰岛素增敏剂为主,临床对磺酰脲类、胰岛素增敏剂的选择倾向较大。     

Factors which determine the accumulation of immunoblasts in gut and skin.

The capacity of immunoblasts from two sources (1) peripheral lymph nodes draining the site of application of a contact sensitizer and (2) mesenteric lymph nodes from mice infected with the gut parasite T. spiralis to migrate to the gut and to inflamed skin sites were compared. The peripheral lymph node blasts readily entered skin sites in a non-specific way but failed to migrate to the gut even when inflammation was induced. By contrast, the mesenteric lymph node blasts readily migrated to the gut in normal mice and in increased amounts to the gut of mice infected with T. spiralis or inflamed with oral turpentine. A small proportion of mesenteric lymph node blasts did, however, migrate, non-specifically to the skin but in much smaller amounts than peripheral lymph node blasts. We conclude that the migration of immunoblasts to the gut has some specificity related to the source from which the cells were taken but little specificity with regard to intraluminal antigen.     

Comparison of sensitizers by detecting reactive oxygen species after photodynamic reaction in vitro.

The production of reactive oxygen species (ROS) has a crucial effect on the result of photodynamic therapy (PDT). Because of this fact, we examined the ROS formation by means of three porphyrin sensitizers (TPPS(4), ZnTPPS(4) and PdTPPS(4)) and compared their effectivity for induction of cell death in the G361 (human melanoma) cell line. The porphyrins used are very efficient water-soluble aromatic dyes with a potential application in photomedicine and have a high tendency to accumulate in the membranes of intracellular organelles such as lysosomes and mitochondria. Interaction between the triplet excited state of the sensitizer and molecular oxygen leads to the production singlet oxygen and other reactive oxygen species to induce cell death. Production of ROS was investigated by molecular probe CM-H(2)DCFDA. Our results demonstrated that ZnTPPS(4) induces the highest ROS production in the cell line compared to TPPS(4) and PdTPPS(4) at concentrations of 1, 10, and 100 microM and light dose of 1 J cm(-2). We also observed a consequence between ROS production and cell survival. In conclusion, these results demonstrate that photodynamic effect depends on sensitizer type, its concentration and light dose.     

Photodynamic therapy with zinc-tetra(p-sulfophenyl)porphyrin bound to cyclodextrin induces single strand breaks of cellular DNA in G361 melanoma cells.

The basis of photodynamic therapy (PDT) is the phototoxicity resulting from co-action of light, sensitizer and oxygen. In this study we demonstrate in vitro phototoxicity measurement on G361 cell lines using ZnTPPS(4) sensitizer bound to cyclodextrin hpbetaCD. We have proved its photodamage effect on cancer cell lines in the visible region of spectrum. We used the halogen lamp (24V/250W) as a source of radiation. After 24h incubation of cell cultures with 10 microM ZnTPPS(4) and 1mM cyclodextrine hpbetaCD, the cells were irradiated for 7.5 min at the total irradiation dose of 12.5 Jcm(-2). Analysis of DNA damage in the cell line after PDT was proved by comet assay and using inversion fluorescent microscope with image analysis. This treatment method gave rise to DNA damage. The used radiation dose of visible light in the absence of sensitizers does not induce DNA breaks in tumour cells. In conclusion, binding of ZnTPPS(4) sensitizer to cyclodextrin hpbetaCD may improve the efficacy of PDT for the treatment of malign melanoma.     

Distribution of crushing strength of tablets.

The distribution of a given set of data is important since most parametric statistical tests are based on the assumption that the studied data are normal distributed. In analysis of fracture mechanics the Weibull distribution is widely used and the derived Weibull modulus is interpreted as a material constant. However, the estimation of this parameter is laborious and subject to estimation problems. It is shown that the Weibull modulus is inherently connected to the coefficient of variation and that the information obtained from the modulus is unclear. The distribution of crushing strength data from nine model tablet formulations and four commercial tablets are shown to follow the normal distribution. The importance of proper cleaning of the crushing strength apparatus is demonstrated.     

Amended final report on the safety assessment of ethyl methacrylate

Ethyl Methacrylate is a methacrylate ester used as a chemical additive in artificial fingernail enhancement products. These products may be applied by trained professionals or be provided directly to consumers with instructions for use. Ethyl Methacrylate readily polymerizes and rapidly reacts with multifunction methacrylates to form a highly cross-linked polymer. The oral LD(50) of Ethyl Methacrylate for rats ranged from 12.7 to 18.14 g/kg. In acute studies with rats, hemoglobinuria and respiratory tract lesions were observed. Animal studies indicate that Ethyl Methacrylate is a skin irritant and sensitizer. In some cases the results were dependent on the vehicle. Evidence of embryotoxicity and teratogenicity were observed in rats injected intraperitoneally with 0.1223 to 0.4076 ml/kg Ethyl Methacrylate. Positive evidence of mutagenicity was observed in the L5178Y mouse lymphoma cell assay, but not in two Ames tests. Case reports cite examples of individuals suffering allergic contact dermatitis from exposure to Ethyl Methacrylate and related methacrylates, and some degree of cross-reactivity appears to exist between widely used acrylates and methacrylates. Information from several clinical registries of sensitization reactions to various agents reported that Ethyl Methacrylate is a sensitizer, but not a potent one. Because Ethyl Methacrylate monomer is short-lived in the normal course of using artificial fingernail-enhancement products, the primary hazard is expected to be inadvertant skin contact. In order to avoid sensitization, it is necessary to avoid skin contact. It is recommended that fingernail-enhancement products containing Ethyl Methacrylate include directions to avoid skin contact because of the sensitizing potential. Based on the available data on the formulation of nail products containing this ingredient, it is concluded that Ethyl Methacrylate is safe as used, when application is accompanied by directions for use as above.     

Non-enzymatic glutathione reactivity and in vitro toxicity: a non-animal approach to skin sensitization.

The development of non-animal methods to predict the potential of chemicals to cause skin sensitization is of great importance. On the basis of many published studies into the underlying chemical mechanisms skin sensitization, the immunological priming which leads to the disease allergic contact dermatitis, is recognized as a reactive chemistry endpoint. Consequently, the combination of chemical assays with in vitro techniques may provide a useful surrogate to animal testing for skin sensitization. This study attempts to investigate the relationship between skin sensitization assessed in the local lymph node assay (LLNA) initially and a thiol reactivity index based on glutathione (GSH), pEC(50) thiol (EC(50) being defined as the concentration of the test substance which gives 50% depletion of free thiol under standard conditions) in combination with a measure of cytotoxicity (pIGC(50)) to Tetrahymena pyriformis (TETRATOX). The pEC(50) thiol values and the pIGC(50) values were determined for twenty-four compounds for which LLNA test data were available. Thiol reactivity was found to discriminate sensitizers from non-sensitizers according to the rule: pEC(50) thiol>-0.55 indicates that the compound will be a skin sensitizer. However, because of metabolic activation a pEC(50) thiol<-0.55 does not necessarily mean that the compound will be a non-sensitizer. Excess toxicity to T. pyriformis (i.e. the extent of toxic potency over that expected by non-polar narcosis) was determined in order to assess biological reactivity. The best discrimination based on excess toxicity in the TETRATOX assay was given by the "rule": excess toxicity>0.50 indicates that the compound will be a skin sensitizer. These approaches become more powerful when combined. When taken together, the thiol and TETRATOX assays predict the sensitization potential of 23 of the 24 compounds correctly. alpha-Hexylcinnamic aldehyde is incorrectly predicted to be a non-sensitizer, whereas LLNA results suggest it may be a weak sensitizer, this inaccuracy being rationalized in terms of its high hydrophobicity. Due to the selectivity of electro(nucleo)philic reactions some sensitizing compounds will not be identified using a single nucleophile such as thiol.     

C57BL/6 mice are resistant to acute restraint modulation of cutaneous hypersensitivity.

C57BL/6 mice, in contrast to BALB/c mice, display minimal behavioral changes in response to environmental stressors and are considered relatively stress-resistant. We have shown that application of acute restraint prior to chemical challenge enhanced cutaneous hypersensitivity (CHS) in BALB/c mice and that this enhanced response is partially glucocorticoid dependent. Due to strain differences in the immune response and in the response to environmental stressors, we hypothesized that acute restraint would not enhance CHS in the less stress-sensitive C57BL/6 mice. We sensitized and challenged C57BL/6 mice with the contact sensitizer, 2, 4-dinitrofluorobenzene (DNFB) in the presence and absence of restraint. Acute restraint, applied prior to chemical challenge, significantly increased serum corticosterone, but to concentrations approximately 60% of those reported for BALB/c mice. Neither restraint nor the exogenous administration of corticosterone enhanced chemical-induced ear swelling in C57BL/6 mice. Pharmacological interruption of the hypothalamic pituitary adrenal axis (HPAA) with the glucocorticoid type II receptor antagonist, RU486, did not alter the development of CHS, however, adrenalectomized (ADX) mice exhibited decreased ear swelling, a measurement that was decreased further by restraint. Combined application of acute restraint and corticosterone prior to chemical challenge significantly enhanced the ear swelling response in C57BL/6 wild-type mice. These data confirm that C57BL/6 mice have a blunted corticosterone response to restraint and that acute restraint does not modulate cutaneous hypersensitivity. Furthermore, our data demonstrate that stress-resistance is not conferred exclusively through the glucocorticoid pathways.     

Acetonitrile serum concentrations and cyanide blood levels in a case of suicidal oral acetonitrile ingestion.

Acute acetonitrile toxicity is mainly dependent on the release of cyanide via hepatic metabolism. Although evaluated in animals, few data are available concerning the toxicokinetic parameters of acetonitrile and acetonitrile-liberated cyanide in human. This paper reports a case of suicidal oral acetonitrile ingestion of about 5 mL without severe symptoms of intoxication in a previously healthy adult male with a body weight of 60 kg. Acetonitrile serum concentrations as well as cyanide blood levels were determined over the whole hospitalization. The elimination half-lives calculated from these data were 32 h for acetonitrile and 15 h for cyanide. After sodium thiosulfate bolus application, the cyanide blood level rapidly decreased to 10% of the initial value, indicating that sodium thiosulfate sufficiently detoxifies acetonitrile-liberated cyanide. Since cyanide levels again increased to maximal values about 4.5 h after sodium thiosulfate application, continued thiosulfate therapy is required as predicted by the long elimination half-lives of acetonitrile and acetonitrile-liberated cyanide. Determination of cyanide and acetonitrile concentrations is recommended for the estimation of optimal individual sodium thiosulfate dosage.     

Contact sensitizers specifically increase MHC class II expression on murine immature dendritic cells

Contact sensitivity is a T-cell-mediated immune disease that can occur when low-molecular-weight chemicals penetrate the skin. In vivo topical application of chemical sensitizers results in morphological modification of Langerhans cells (LC). Moreover, within 18 h, LC increase their major histocompatibility complex (MHC) class II antigens expression and migrate to lymph nodes where they present the sensitizer to T lymphocytes. We wanted to determine if such an effect could also be observed in vitro. However, because of the high genetic diversity encountered in humans, assays were performed with dendritic cells (DC) obtained from a Balb/c mouse strain. The capacity of a strong sensitizer, DNBS (2,4-dinitrobenzene sulfonic acid), to modulate the phenotype of bone marrow-derived DC in vitro, was investigated. A specific and marked increase of MHC class II molecules expression was observed within 18 h. To eliminate the use of animals in sensitization studies, the XS52 DC line was tested at an immature stage. A 30-min contact with the strong sensitizers DNBS and oxazolone, or the moderate mercaptobenzothiazole, resulted in upregulation of MHC class II molecules expression, analyzed after 18-h incubation. This effect was not observed with irritants (dimethyl sulfoxide and sodium lauryl sulfate) nor with a neutral molecule (sodium chloride). These data suggested the possibility of developing an in vitro model for the identification of the sensitizing potential of chemicals, using a constant and non animal-consuming material.     

The acute toxicity of 2-benzylaminopyridine.

2-Benzylaminopyridine (2-BAP) has been suspected of causing adverse reactions in man including headache, malaise, and skin rash. The purpose of this investigation was to determine and partially characterize the acute toxicity of 2-BAP. In mice the estimated LD50 of 2-BAP dissolved in equimolar HCl was 1187 mg/kg po and 224 mg/kg ip. Daily administration of 600 mg/kg po for 30 days indicated no significant cumulative toxicity. Disappearance from whole mice was determined following 50 mg/kg ip. The half-life calculated from the first order disappearance rate constant was found to be 15 min. In mice, pretreatment with 2-BAP (50 mg/kg ip) significantly prolonged the loss of righting reflex induced by ethanol (4 g/kg ip). Guinea pigs were used to study the dermal irritant and sensitizing properties of 2-BAP. Acute exposure of the bare skin to a 2% solution resulted in no primary irritation. 2-BAP solution was applied on Days 1, 3, and 5. Three weeks later a challenge application to a different area of skin resulted in a slight erythema indicating that 2-BAP may be a dermal sensitizer.     

Validity of methods to predict the respiratory sensitizing potential of chemicals: A study with a piperidinyl chlorotriazine derivative that caused an outbreak of occupational asthma.

A piperidinyl chlorotriazine (PCT) derivative, used as a plastic UV-stabilizer, caused an outbreak of occupational asthma. We verified, in BALB/c mice, the sensitizing potential of PCT in comparison to a known respiratory sensitizer (toluene diisocyanate [TDI]) and a known dermal sensitizer (oxazolone), using three different methods in order to evaluate the validity of current models of sensitization. These included the local lymph node assay (LLNA) and the mouse IgE test. In addition, respiratory hyper-reactivity was assessed following a novel protocol involving dermal sensitization (20 microl of a 3% solution on each ear for three days) and intranasal challenge (0.1% or 1%, 10 microl per nostril on day 10), followed, after 24 h, by a methacholine challenge (using whole-body plethysmography), bronchoalveolar lavage, and histology. PCT was also used for structure-activity relationship (SAR) models for (respiratory) sensitization. High concentrations of PCT (10 and 20%) resulted in significant responses in the local lymph node assay (LLNA; stimulation indices (SI) of 2.7 +/- 0.9 and 3.2 +/- 0.6, respectively). The mouse IgE test was positive with 20% PCT only. Methacholine responsiveness was increased only in previously sensitized mice receiving a challenge with TDI or PCT. However, there was no evidence for pulmonary inflammation. The SAR studies indicated that PCT could be a respiratory sensitizer. Based on an approved test protocol such as the LLNA and the mouse IgE test, PCT proved to be a weak sensitizer when compared to TDI and oxazolone. However, in a protocol involving an intranasal challenge, PCT appeared to be a respiratory sensitizer of similar potency to TDI.     

Application of a generic physiologically based pharmacokinetic model to the estimation of xenobiotic levels in human plasma.

Estimation of xenobiotic kinetics in humans frequently relies upon extrapolation from experimental data generated in animals. In an accompanying paper, we have presented a unique, generic, physiologically based pharmacokinetic model and described its application to the prediction of rat plasma pharmacokinetics from in vitro data alone. Here we demonstrate the application of the same model, parameterized for human physiology, to the estimation of plasma pharmacokinetics in humans and report a comparative evaluation against some recently published predictive methods that involve scaling from in vivo animal data. The model was parameterized through an optimization process, using a training set of in vivo data taken from the literature, and validated using a separate test set of published in vivo data. On average, the vertical divergence of the predicted plasma concentrations from the observed data, on a semilog concentration-time plot, was 0.47 log unit. For the training set, more than 80% of the predicted values of a standardized measure of the area under the concentration-time curve were within 3-fold of the observed values; over 70% of the test set predictions were within the same margin. Furthermore, in terms of predicting human clearance for the test set, the model was found to match or exceed the performance of three published interspecies scaling methods, all of which showed a distinct bias toward overprediction. We conclude that the generic physiologically based pharmacokinetic model, as a means of integrating readily determined in vitro and/or in silico data, is potentially a powerful, cost-effective tool for predicting human xenobiotic kinetics in drug discovery and risk assessment.     

A population analysis of the pharmacokinetics and pharmacodynamics of midazolam in the rat.

The concentration-EEG effect relationship of midazolam in the rat was studied from a population perspective. Plasma concentration and EEG effect data from 27 rats were available for analysis. Effect parameters derived from aperiodic EEG analysis were used as effect parameters. The population analysis gave results that were similar to the sample theory estimates (means s and SDs) obtained from the fits to individual data sets. Reanalysis of the EEG data using mean population pharmacokinetic parameters as input to the pharmacodynamic model led to poorer estimation of the pharmacodynamic parameters: particularly EC50. Inclusion of one observed plasma concentration per individual significantly improved the estimation of the pharmacodynamic parameters and led to results that were virtually indistinguishable from those obtained using complete pharmacokinetic data.     

Residues of lindane and DDT in drugs from wild medicinal plants in a cultivated forest. 2

The results of testing the persistency of Lindane and DDT in wild medicinal plants collected in a forest after application of bercema-Aero-Super are demonstrated. They illustrate, that after the application the herbs from the first year of vegetation will not meet the state quality requirements. An estimation is given about the toxicological relevance of the pesticide residues in herbs which are used for tea infusions.     

Estimation of agonist affinity, using the slope of the concentration-effect curve: comment on the method of Calderone and Martinotti

Recently, Calderone and Martinotti (J Pharmacol Toxicol Meth40:57–62, 1998) presented a simple equation for the estimation of agonist dissociation equilibrium constants from data obtained from receptor inactivation experiments using an irreversible, competitive antagonist. In the present paper, however, it is demonstrated that application of this equation will result in significant over- and underestimation of agonist affinity in the case of flat and steep concentration-effect curves, respectively. Therefore, accurate estimation of agonist affinity using irreversible receptor inactivation requires detailed curve-shape information from both control and antagonist-treated tissues.