富氢液对脓毒血症大鼠肠黏膜屏障保护作用及机制研究

目的探讨富氢液对脓毒血症大鼠肠黏膜屏障的影响及其可能机制。方法清洁级SD大鼠30只,随机分为假手术组(sham组)、脓毒血症组(SEP组)及富氢液处理组(C组),每组10只。sham组仅开腹,显露盲肠;C组和SEP组行盲肠结扎穿孔术,再分别静脉给予富氢液和等量生理盐水。3组大鼠均于模型建立4 h后处死,观察小肠组织病理学结果;酶联免疫吸附法(ELISA)检测肠脂肪酸结合蛋白(I-FABP)、D-乳酸、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)浓度;蛋白质印迹(Western-blot)法检测B淋巴细胞瘤-2基因(Bcl-2)、Bax、Caspase-3蛋白表达水平。结果光镜下sham组未出现病理损伤,SEP组损伤较重,C组损伤较轻。与sham组比较,SEP组I-FABP、D-乳酸升高[(817.12±50.39)ng/ml比(378.54±40.50)ng/ml;(513.84±42.91)ng/ml比(329.98±38.11)ng/ml,P<0.05]。与SEP组比较,C组I-FABP、D-乳酸降低[(548.92±42.19)ng/ml比(817.12±50.39)ng/ml;(437.89±36.12)ng/ml比(513.84±42.91)ng/ml,P<0.05]。与sham组比较,SEP组TNF-α、IL-6、IL-10升高[(196.51±30.31)ng/ml比(75.54±10.21)ng/ml;(68.69±3.39)ng/ml比(20.97±2.38)ng/ml;(149.61±20.21)ng/ml比(80.69±13.51)ng/ml,P<0.05];与SEP组比较,C组TNF-α、IL-6降低,IL-10升高[(118.47±10.14)ng/ml比(196.51±30.31)ng/ml;(45.36±5.14)ng/ml比(68.69±3.39)ng/ml;(178.52±26.31)ng/ml比(149.61±20.21)ng/ml,P<0.05]。与sham组比较,SEP组Bcl-2蛋白表达升高,Bax与Caspase-3降低(P<0.05);与SEP组比较,C组Bcl-2蛋白表达降低,Bax与Caspase-3升高(P<0.05)。结论富氢液对脓毒血症大鼠肠黏膜屏障有保护作用,其机制与抑制凋亡相关因子有关。     

高渗环境对假体周围感染生物膜形成影响的实验研究

目的探讨高渗环境对关节置换术后假体周围感染中表皮葡萄球菌生长和生物膜形成影响。方法用新西兰大白兔建立假体表皮葡萄球菌感染的动物模型,分为实验组和对照组各15只。实验组注入细菌的同时注入40g/L的氯化钠溶液0.1mL,对照组注入生理盐水0.1mL。分别于注入细菌后2、4、6、8和16d对假体感染动物行关节液细菌计数、假体周围组织学观察和假体扫描电镜观察。结果注入细菌后2-4d实验组和对照组关节液细菌计数差异无显著性（P〉0.05）,注入细菌后6、8和16d两组细菌计数差异有统计学意义（t=2.16-3.17,P〈0.05）。注入细菌后4d实验组动物假体周围组织均有中性粒细胞浸润;对照组注入细菌后4d开始出现炎细胞浸润,注入细菌后6d所有动物假体周围组织中均有炎细胞浸润。两组注入细菌后4d均观察到假体表面有少量细菌黏附;注入细菌后6、8和16d实验组假体表面黏附的细菌较对照组增多。结论关节置换术后假体周围高渗环境能够促进表皮葡萄球菌的生长以及生物膜的形成。     

Blood-Brain Barrier Permeability of Asiaticoside,Madecassoside and Asiatic Acid in Porcine Brain Endothelial Cell Model

Neurotherapeutic potentials of Centella asiatica and its reputation to boost memory, prevent cognitive deficits and improve brain functions are widely acknowledged. The plant's bioactive compounds, i.e. asiaticoside, madecassoside and asiatic acid were reported to have central nervous system (CNS) actions, particularly in protecting the brain against neurodegenerative disorders. Hence, it is important for these compounds to cross the blood-brain barrier (BBB) to be clinically effective therapeutics. This study aimed to explore the capability of asiaticoside, madecassoside and asiatic acid to cross the BBB using in vitro BBB model from primary porcine brain endothelial cells (PBECs). Our findings showed that asiaticoside, madecassoside and asiatic acid are highly BBB permeable with apparent permeability (P_app) of 70.61 ± 6.60, 53.31 ± 12.55 and 50.94 ± 10.91 × 10^?6 cm/s respectively. No evidence of cytotoxicity and tight junction disruption of the PBECs were observed in the presence of these compounds. Asiatic acid showed cytoprotective effect towards the PBECs against oxidative stress. This study reported for the first time that Centella asiatica compounds demonstrated high capability to cross the BBB, comparable to central nervous system drugs, and therefore warrant further development as therapeutics for the treatment of neurodegenerative diseases.     

Pollen shells and soluble factors play non-redundant roles in the development of allergic conjunctivitis in mice

PurposeWe aimed to clarify the role of particulate allergen exposure to the conjunctiva in the development of allergic conjunctivitis.MethodsWe administered ragweed pollen suspension, pollen extract, pollen shell, particulate air pollutants, and their combinations to the mouse conjunctiva five days a week without prior sensitization. Clinical signs were scored. Histological changes, cellular infiltrations, mRNA expressions, lymph node cell recall responses, and serum immunoglobulin levels were assessed. Immune cell-depleting antibodies and ST2 knockout mice were used to investigate the cellular and molecular requirements.ResultsPollen suspension, but not the extract or shell alone, induced robust eosinophilic conjunctivitis, accompanied by a proliferative response of epithelial cells. A combination of pollen extract and shell completely restored eosinophil accumulation. In addition, eosinophilic conjunctivitis was induced by a mixture of particulate air pollutants and pollen extract. Mechanistically, eosinophil accumulation was ameliorated by deficiency of the IL-33 receptor ST2 and abolished by depleting CD4⁺ T cells. Pollen shells, but not the extract, induced IL-33 release from conjunctival epithelial cells in vivo.ConclusionsOur results indicate the non-redundant roles for the allergens’ particulate properties and soluble factors in the development of allergic conjunctivitis.     

Cross-neutralization between vaccine and circulating wild-type mumps viruses in Korea

Mumps is a contagious disease caused by the mumps virus. It can be prevented using mumps vaccines, administered as a measles-mumps-rubella (MMR) vaccine. For first and second dose immunization, children aged 12–15 months and 4–6 years have been administered this vaccine since 1997 in Korea. Nevertheless, mumps outbreaks still occur in vaccinated populations worldwide. Hence, immunity against these diseases may be attenuated, or there are antigenic differences between currently available vaccine strains and circulating wild-type viruses. After the introduction of national immunization programs in Korea, mumps cases became sporadic. Viral genotypes F, H, and I have emerged since 1998 whereas the vaccine strains belong to genotype A. Here, we compared the amino acid sequences of the haemagglutinin-neuraminidase (HN) gene from wild-type viruses and the mumps vaccine and measured the cross-neutralization titers between them. We selected the F, H, and I wild-type mumps strains circulating in Korea from 1998 to 2016 and analyzed changes in the amino acid sequence of the protein encoded by the HN gene. We measured mumps virus-specific IgG and rapid focus reduction neutralization test (FRNT) titers in Korean isolates and sera obtained from 50 children aged 1–2 years who had been administered a single dose of MMR vaccine. Analysis of the HN protein sequences disclosed no changes in the glycosylation sites but did reveal 4–5 differences between the Korean isolates and the genotype A vaccine strain in terms of the neutralizing epitope sites on their HN proteins. Post-vaccination FRNT titers were significantly lower against genotypes F, H, and I than they were against genotype A. This finding highlights the possibility of a recurrence of mumps outbreaks in vaccinated populations depending on the degree of genetic conservation of the HN gene. Further research into this issue is needed to prevent the resurgence of mumps.     

Rociletinib (CO-1686) enhanced the efficacy of chemotherapeutic agents in ABCG2-overexpressing cancer cells in vitro and in vivo

Overexpression of adenosine triphosphate (ATP)-binding cassette subfamily G member 2 (ABCG2) in cancer cells is known to cause multidrug resistance (MDR), which severely limits the clinical efficacy of chemotherapy. Currently, there is no FDA-approved MDR modulator for clinical use. In this study, rociletinib (CO-1686), a mutant-selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), was found to significantly improve the efficacy of ABCG2 substrate chemotherapeutic agents in the transporter-overexpressing cancer cells in vitro and in MDR tumor xenografts in nude mice, without incurring additional toxicity. Mechanistic studies revealed that in ABCG2-overexpressing cancer cells, rociletinib inhibited ABCG2-mediated drug efflux and increased intracellular accumulation of ABCG2 probe substrates. Moreover, rociletinib, inhibited the ATPase activity, and competed with [¹²⁵I] iodoarylazidoprazosin (IAAP) photolabeling of ABCG2. However, ABCG2 expression at mRNA and protein levels was not altered in the ABCG2-overexpressing cells after treatment with rociletinib. In addition, rociletinib did not inhibit EGFR downstream signaling and phosphorylation of protein kinase B (AKT) and extracellular signal-regulated kinase (ERK). Our results collectively showed that rociletinib reversed ABCG2-mediated MDR by inhibiting ABCG2 efflux function, thus increasing the cellular accumulation of the transporter substrate anticancer drugs. The findings advocated the combination use of rociletinib and other chemotherapeutic drugs in cancer patients with ABCG2-overexpressing MDR tumors.     

鼻内镜术后不同冲洗液冲洗鼻腔的效果对比

目的:观察鼻渊通窍颗粒稀释液与生理盐水对鼻内镜术后鼻腔冲洗的效果。方法:临床纳入进行功能性内窥镜鼻窦手术患者90例,研究组给予鼻渊通窍颗粒稀释液,对照组给予生理盐水。每周定期对患者进行鼻内窥镜的检查,观察两组患者4周内鼻腔症状以及术腔清洁、囊泡形成、肉芽生成等体征变化情况。结果:两组术后4周冲洗治疗结果表明,两者均有治疗作用,且治疗效果相当,但鼻渊通窍颗粒稀释液冲洗在术后第2周时对术腔痂皮脱落、黏膜水肿、鼻分泌物减少、囊泡形成治疗作用较生理盐水冲洗要好,差异有显著性(P<0.05);研究组治疗4周,总有效率为93.33%,对照组有效率为91.11%,差异无显著性(P>0.05)。结论:鼻渊通窍颗粒稀释液能够有效清洁术后的鼻腔,具有减轻鼻腔黏膜水肿、囊泡再生以及促进黏膜功能恢复等作用。     

Toxicology of 3-epi-deoxynivalenol,a deoxynivalenol-transformation product by Devosia mutans 17-2-E-8

Microbial detoxification of deoxynivalenol (DON) represents a new approach to treating DON-contaminated grains. A bacterium Devosia mutans 17-2-E-8 was capable of completely transforming DON into a major product 3-epi-DON and a minor product 3-keto-DON. Evaluation of toxicities of these DON-transformation products is an important part of hazard characterization prior to commercialization of the biotransformation application. Cytotoxicities of the products were demonstrated by two assays: a MTT bioassay assessing cell viability and a BrdU assay assessing DNA synthesis. Compared with DON, the IC₅₀ values of 3-epi-DON and 3-keto-DON were respectively 357 and 3.03 times higher in the MTT bioassay, and were respectively 1181 and 4.54 times higher in the BrdU bioassay. Toxicological effects of 14-day oral exposure of the B6C3F₁ mouse to DON and 3-epi-DON were also investigated. Overall, there were no differences between the control (free of toxin) and the 25 mg/kg bw/day or 100 mg/kg bw/day 3-epi-DON treatments in body and organ weights, hematology and organ histopathology. However, in mice exposed to DON (2 mg/kg bw/day), white blood cell numbers and serum immunoglobulin levels were altered relative to controls, and lesions were observed in adrenals, thymus, stomach, spleen and colon. Taken together, in vitro and in vivo studies indicate that 3-epi-DON is substantially less toxic than DON.     

心肾综合征低钠血症患者抗利尿激素测定及高渗盐水强化利尿的疗效

目的:探讨抗利尿激素(ADH)在心肾综合征(CRS)伴低钠血症中的作用及高渗盐水强化利尿(HS aggDiu)对心肾功能和ADH释放的影响。方法:CRS伴低钠血症患者25例,测定其血浆ADH水平。将患者随机分为(HS aggDiu)13例和单纯强化利尿组(aggDiu组)12例,分别给予高渗盐水并大剂量呋塞米静脉滴注和单纯大剂量呋塞米静脉滴注,记录2组患者心肾功能、水钠潴留和血钠的变化。结果:HS aggDiu组尿量增加、体重减轻、血肌酐下降、NYHA心功能分级改善等方面显著改善、水钠潴留减轻,血钠水平升高,其疗效显著优于aggDiu组。同时,血ADH水平并未随着血钠和渗透压的恢复而明显升高,说明HS aggDiu减少了ADH的非渗透性释放。结论:CRS伴低钠血症与ADH的非渗透性释放有关;可有效减轻水钠潴留,改善心肾功能,并减轻ADH的非渗透性释放。