Neuraminidase- and benzalkonium chloride-dependent inhibition of basic peptide-induced rat mast cell secretion

Two types of inhibition of basic peptide-induced rat mast cell secretion are reported. Pretreatment of rat peritoneal mast cells with Vibrio comma neuraminidase, an enzyme which cleaves sialic acid from oligosaccharides, led to inhibition of 5-hydroxytryptamine release induced by the basic peptides polylysine, corticotropin_1–24 and a decapeptide sequence of human IgE. Inhibition was similarly observed when mast cells were challenged in the presence of the cationic cell membrane-active substance benzalkonium chloride. It is postulated that both of these experimental procedures inhibit basic peptide-induced secretion by depletion of cell surface negative charge. Sialic acid itself does not act as a specific receptor for basic peptides, since a molar excess of sialic acid in free solution failed to inhibit secretion by binding to basic peptides in the fluid phase.     

苯扎氯铵改性粘接剂对牙本质-树脂远期粘接强度的影响研究

目的:评估苯扎氯铵(benzalkonium chloride,BAC)改性粘接剂对牙本质-树脂远期粘接强度的影响.方法:将BAC加入到美国3M第八代全酸蚀树脂粘接剂(single bond universal,SBU)中,制成不同浓度的改性粘接剂.将保存于百里香酚溶液中的100颗第三磨牙随机分为10组(n=10),A...     

HPLC法测定吡嘧司特钾滴眼液中苯扎溴铵和苯扎氯铵的含量

摘要：目的建立HPLC法测定吡嘧司特钾滴眼液中苯扎溴铵和苯扎氯铵的含量。方法色谱柱：C18（4．6mm×150mm,5μm）;流动相：0．02mol·L^-1庚烷磺酸钠溶液（含0．1％三乙胺,用磷酸调节PH为3．45±0．1）-乙腈（35：65）;检测波长：210nm;柱温：40℃。结果苯扎溴铵在49．86～997．2ng范围内线性关系良好（r=0．9999）,最小检出量为0．15ng,平均回收率为97．4％,RSD为1．0％;苯扎氯铵在9．912-24．78ng范围内呈良好的线性关系（r=0．9999）,最小检出量为0．85ng,平均回收率为99．9％,RSD为1．0％。结论本方法简便、快速、准确、灵敏度高、重复性好,可用于吡嘧司特钾滴眼液中苯扎溴铵和苯扎氯铵的含量测定。     

Topical hyaluronan alone promotes corneal epithelial cell migration whereas combination with benzalkonium chloride impairs epithelial wound healing

Abstract

Purpose: To evaluate the effects of topical hyaluronan (HA) on corneal epithelial wound healing when administered with or without benzalkonium chloride (BAC).

Methods: A cultured human corneal epithelial cell line (HCE-T) was subjected to in vitro scratch assays and in situ epithelial migration was evaluated in organ-cultured rabbit corneas. The corneal epithelium of C57BL/6J mice was also evaluated to determine in vivo wound healing. An in vivo imaging system was also used to evaluate the effects of HA on eye drop retention on the ocular surface.

Results: The findings revealed the promotion of HCE-T migration, in situ rabbit corneal epithelial migration, and in vivo wound healing in mouse corneal epithelium by HA. Pre-treatment with HA also protected against delayed epithelial wound healing in BAC in vitro. However, pre-treatment with 3?mg/mL HA did not show a protective effect against BAC in vivo, but instead delayed epithelial wound healing and increased detection of cleaved caspase-3. This suggested that HA promotes the retention of BAC on the ocular surface. The instilled HA was retained after 15?min, at a significantly higher rate than for phosphate-buffered saline.

Conclusions: The combination of HA and BAC impaired wound healing in the corneal epithelium.     

阿奇霉素滴眼液抑菌剂筛选与抑菌效力研究

目的:探索阿奇霉素滴眼液中抑菌剂的合理添加剂量。方法:根据药物成分的特性,选择若干适宜的抑菌剂,并对不同配比浓度抑菌剂的抑菌效果进行初步考察,筛选出适宜的供试品抑菌剂加入量。选择有代表性的挑战微生物加入供试品中进行挑战试验,测试微生物在不同时间段的存活情况。结果:0.003%的苯扎氯铵对5种挑战微生物的生长有良好的抑制作用,且为最低有效剂量。结论:0.003%的苯扎氯铵可作为阿奇霉素滴眼液的抑菌剂。     

复方醋酸地塞米松制剂中9种抑菌剂含量测定方法的建立及评价

目的 建立HPLC同时测定复方醋酸地塞米松乳膏、凝胶中可能含有的9种常见抑菌剂的方法。对2018年国家药品抽验到的国内44个生产厂家的样品进行检测及评价。方法 采用Waters Xbridge Shield RP18色谱柱（4.6 mm×250 mm,5 μm）;以1%三乙胺溶液（加磷酸调节pH至3.0）为流动相A,乙腈为流动相B,进行梯度洗脱;检测波长215 nm。采用该方法对国内样品进行检测,并对该复方制剂中抑菌剂使用进行有效性和安全性评价。结果 在该色谱条件下,9种抑菌剂分离良好,在1~70 μg·ml^-1内线性关系良好,平均回收率为91.2%~100.4%,RSD为0.1%~6.7%。样品中共检测出布罗波尔、羟苯甲酯、羟苯乙酯、羟苯丙酯和苯扎溴铵5种抑菌剂,抑菌剂浓度为0.48~2.89 mg·g^-1。结论 样品中存在抑菌剂使用不规范现象。本研究可为该品种中抑菌剂的规范使用提供技术支持。     

IL6Rα function in myeloid cells modulates the inflammatory response during irritant contact dermatitis

Irritant contact dermatitis (ICD) is characterized by epidermal hyperplasia, infiltration of leucocytes into lesional skin and inflammatory cytokine release. The cellular infiltrate during ICD comprises primarily cells of the myeloid lineage. Our group has previously shown that the cytokine IL‐6 confers a protective effect to lesional skin during ICD. How IL‐6Rα function in myeloid cells is involved in the inflammatory response during ICD is, however, unknown. In the present study, utilizing a chemical model of ICD, it is shown that mice with a myeloid‐specific knockout of the IL‐6Rα (IL‐6Rα^Δmyeloid) display an exaggerated inflammatory response to benzalkonium chloride (BKC) and Jet propellant‐8 (JP8) fuel, two well‐characterized irritants relative to littermate control. Results from immunohistochemical and flow cytometric analyses revealed that IL‐6Rα^Δmyeloid mouse skin displayed increased epidermal hyperplasia and inflammatory monocyte influx into lesional skin but lower numbers of resident macrophages relative to littermate controls after irritant exposure. Multiplex immunoassay revealed significantly higher levels of pro‐inflammatory cytokines IL‐1α and TNF‐α, but reduced expression of chemokine proteins including CCL2‐5, CCL7, CCL11, CXCL1 and CXCL10 in IL‐6Rα^Δmyeloid mouse skin relative to littermate control following irritant exposure. These results highlight a previously unknown role of IL‐6Rα function in myeloid cells in modulating the inflammatory response and myeloid population dynamics during ICD.     

In vitro methods of assessing ocular biocompatibility using THP-1-derived macrophages

Macrophages play an important role in the elimination of infections, the removal of debris and in tissue repair after infection and trauma. In vitro models that assess ocular biomaterials for toxicity typically focus on the effects of these materials on epithelial or fibroblast cells. This investigation evaluated known ocular toxins deposited on model materials for their effects on the viability and activation of macrophages. THP-1-derived macrophages were cultured onto silicone films (used as a base biomaterial) deposited with chemical toxins (benzalkonium chloride (BAK), zinc diethyldithiocarbamate (ZDEC) and lipopolysaccharide (LPS)). Utilizing three fluorescent dyes calcein, ethidium homodimer-1 (EthD-1) and annexin V, the viability of macrophages attached to the biomaterial was determined using confocal microscopy. Propidium iodide (PI) staining and alamarBlue® (resazurin) reduction were used to assess cell death and metabolic activity. CD14, CD16, CD33, CD45, and CD54 expression of adherent macrophages, were also evaluated to detect LPS activation of macrophages using flow cytometry. The sensitivity of this test battery was demonstrated as significant toxicity from treated surfaces with ZDEC (0.001–0.01%), and BAK (0.001%–0.1%) was detected. Also, macrophage activation could be detected by measuring CD54 expression after exposure to adsorbed LPS. These in vitro methods will be helpful in determining the toxicity potential of new ocular biomaterials.     

普拉洛芬滴眼液中苯扎氯铵的含量测定方法研究

摘要 目的：建立普拉洛芬滴眼液中苯扎氯铵含量的HPLC测定方法。方法：用十八烷基硅烷键合硅胶为填充剂,以0.07mol?L乙酸铵溶液（含三乙胺1%,冰醋酸调节pH至5.0）-乙腈（25：75）为流动相,检测波长为210nm。结果：在选定的色谱条件下,各组分分离度良好,苄基十二烷基二甲基氯化铵、苄基十四烷基二甲基氯化铵分别在3.4～33.8μg?mL-1,0.7～5.5μg?mL-1的浓度范围内线性关系良好（r>0.999）;精密度试验的RSD分别为0.2%和0.7%、稳定性试验的RSD均不大于1.0%;回收率分别为99.9%,101.2%（n=9,RSD<1.0%）。C12与C14的检出限分别为3.4 ng?mL-1和110.5 ng?mL-1。在测定3批样品中,其中C12含量（μg?mL-1）分别为57.195,57.009,56.830;C14含量（μg?mL-1）分别为12.850,12.714,12.937;苯扎氯铵含量（μg?mL-1）分别为70.045,69.723,69.767。结论：本方法专属性强,准确度高,重现性好,能分别控制普拉洛芬滴眼液中苯扎氯铵两种同系物的含量。     

HPLC法测定氯霉素滴眼液中苯扎溴铵含量

目的：建立HPLC测定氯霉素滴眼液中苯扎溴铵含量的方法。方法：采用苯基硅烷键合硅胶为固定相;流动相为0.05 moL· L^-1磷酸二氢钾溶液（用三乙胺调节pH值至5.5）-乙腈（40∶60）;检测波长220 nm;流量1.2 mL· min^-1;柱温40℃;进样量20μL。结果：苯扎溴铵在0.0304～1.218μg范围内呈良好的线性关系（r＝0.9998）;检测限为3.45 ng,定量限为6.9 ng;平均回收率为99.8％（n＝9）,RSD＝1.5％。结论：该方法准确、可靠,可有效控制氯霉素滴眼液中苯扎溴铵的含量。