参金止痢口服液对腹泻哺乳仔猪肠道微生物菌群影响的差异性分析

研究参金止痢口服液对腹泻哺乳仔猪肠道微生物菌群的影响。选取同等饲养条件下7~15日龄体重相近的“杜×长×大”三元杂交仔猪共计9头,其中有腹泻症状的6头将其随机平均分为腹泻空白组(FXK)和腹泻中药组(FZ),正常的3头为健康空白组(JK),中药组灌服参金止痢口服液,空白组灌服同等体积生理盐水,连续灌胃5d,在给药后的第8天对仔猪采血进行抗体及细胞因子水平测定,并采集结肠内容物提取样本中菌群总DNA进行16S rRNA高通量测序。结果显示：与FXK组相比,FZ组血清中IgG含量显著升高(P＜0.05),IgM含量显著降低(P＜0.05),炎性因子IL-1β、IL-6、TNF-α含量显著下调(P＜0.01)。α、β多样性分析结果表明,三个组结肠肠道菌群结构差异显著。在门分类水平上,FZ组仔猪肠道拟杆菌门相对丰度显著高于FXK组(P＜0.05),变形菌门相对丰度显著低于FXK组(P＜0.05),而FZ与JK组之间两个菌门差异不显著。属水平上,FZ组韦荣球菌属和乳杆菌属相对丰度显著低于FXK组(P＜0.05),普雷沃氏菌属相对丰度显著高于FXK组(P＜0.05),FZ组与JK组之间这三个菌属差异不显著。综上结果表明,参金止痢口服液可能通过调节免疫及炎性相关因子水平和调节肠道菌群结构从而发挥治疗哺乳仔猪腹泻的作用。     

Passive immunisation of fish against vibriosis, comparison of intraperitoneal, oral and immersion routes

Passive immunisation of fish was conducted to determine whether anti-Vibrio anguillarum whole sera (AVA) and affinity-purified AVA raised in sheep, rabbits and rainbow trout (Oncorhynchus mykiss) were persistent when injected and orally administered into rainbow trout. These responses were compared with active immunisation by immersion in, and intraperitoneal (i.p.) injection with, formalin-killed V. anguillarum cells. Sheep and rabbit AVA were detected in rainbow trout sera for up to 70 days (half-life 21 days) after i.p. injection as determined by an enzyme-linked immunosorbent assay (ELISA). The relative percentage survival (RPS) of passively immunised rainbow trout challenged with virulent V. anguillarum after an injection was comparable to that of active immunisation by immersion after 1 month post-immunisation (p.i). Affinity-purified sheep and rabbit AVA exhibited the same protective potential as whole serum in rainbow trout. Rabbit and sheep immune sera diluted 1:8 and 1:50, respectively, provided equivalent protection as undiluted fish immune serum. An active immune response against passively acquired heterologous immunoglobulins was demonstrated by ELISA, with responses against sheep AVA being less than those against rabbit AVA. Rainbow trout given purified sheep AVA conjugated to LTB (the GM-1-binding subunit of Escherichia coli heat-labile toxin) and administered orally had an RPS of 37.5% at 15 days and 27% at 1 month p.i. In contrast, fish given sheep AVA conjugated to TraT (an internal membrane of E. coli) or in micellar form with Quil-A had RPSs of only 18.7 and 6.2%, respectively, after 15 days, and 13.3 and 0% after 1 month, respectively. The protection conferred by immune sera was shown to be due to the immunoglobulin component alone. Heat inactivation of the complement in sera had no effect on the potency of immune sera.     

The effect of bentonite concentration on the drug delivery efficacy of a pH-sensitive alginate/bentonite hydrogel

We produced a protein loaded, pH-sensitive alginate-bentonite hydrogel for wound dressings. Alginate is a nontoxic polysaccharide with favorable pH-sensitive properties that make it useful for the intestinal delivery of protein drugs. However, the use of alginate for drug delivery is limited by drug leaching and rapid dissolution of alginate at the higher pH, which may result in lower entrapment efficiency and a burst in the release of entrapped protein drugs. To overcome these problems, we created a novel cross-linked alginate-bentonite hydrogel by combining mineral-rich bentonite with the alginate matrix along with an additive to ensure controlled release. We analyzed the gel in the drug loading process in an aqueous environment by looking at the release profiles of a model protein drug (BSA) from the hydrogel at pH values of 4.5, 5.2 (skin area) and 7.4, 9.2 (wound area). The swelling ratio decreased with bentonite concentration, but did not fall below 6. The rate of drug release was slowest at a pH value of 4.5 and fastest at a pH value of 9.2. The rate of drug release decreased with bentonite concentration. The presence of bentonite prevents the rapid dissolution of alginate at the higher pH, ensuring the controlled release of the entrapped drug.     

苯甲酰基脲类和肼类昆虫生长调节剂对棉铃虫的生物活性

研究了作用机制完全不同的苯甲酰基脲类（氟铃脲）和双苯甲酰基肼类（抑食肼和虫酰肼）昆虫生长调节剂（ＩＧＲ）及其混用对棉铃虫的生物活性和作用方式。供试３龄幼虫经氟铃脲点滴处理后,逐渐表现活动缓慢、反应迟钝、取食减少,最终死亡。死亡幼虫体壁薄而易破,头部背面呈现明显突起的白色透明水泡状（易破体并有体液流出）,部分虫体黑化,表现明显的蜕皮障碍。３龄幼虫经抑食肼处理后,最先也表现为行为迟缓、食量减小,最终死亡的幼虫虫体缩小并发黑,部分存活的幼虫２ｄ后即蜕皮。氟铃脲与抑食肼混合处理后,部分试虫呈现氟铃脲的中毒症状,部分试虫则表现抑食肼中毒症状,少量个体两种症状并发。氟铃脲对棉铃虫３幼虫的ＬＤ５０值（点滴法）和１龄幼虫的ＬＣ５０值（叶片法）分别为０．７４３４μｇａ．ｉ．／头和２．０５９２ｍｇ／Ｌ,表现出很强的触杀作用和胃毒作     

Preparation and evaluation of self-microemulsifying drug delivery system of baicalein

The main object of this work is to prepare self-microemulsifying drug delivery system (SMEDDS) for oral bioavailability enhancement of a poorly water-soluble drug, baicalein. SMEDDS is the mixture of surfactants, cosurfactants, and oils, which are emulsified in aqueous media under conditions of gentle agitation or gastrointestinal motility. Solubility of baicalein was determined in various vehicles. Pseudo-ternary phase diagrams were constructed to identify the efficient self-emulsification region and droplet size distributions of the resultant microemulsions were determined using a particle size analyzer. Optimized SMEDDS formulations for baicalein were Cremophor RH40 (53.57%) as surfactant, Transcutol P (21.43%) as cosurfactant, and Caprylic capric triglyceride (ODO, 25%) as oil. The drug release rate of SMEDDS was significantly higher than that of the baicalein suspension. Comparison of the pharmacokinetics between baicalein-loaded SMEDDS and baicalein suspension was also performed in rats. The plasma concentrations of baicalein and baicalin, its mainly conjugated metabolite, were determined by HPLC method. The in vivo results showed that the absorption of baicalein from SMEDDS resulted in about 200.7% increase in relative bioavailability compared with that of the baicalein suspension. Our studies illustrated the potential use of SMEDDS for the delivery of hydrophobic compounds, such as baicalein by the oral route.     

FAM3C activates Akt to promote viability of oral squamous-cell carcinoma cells

AIM: To investigate the expression and roles of family with sequence similarity 3, member C (FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte (DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with siFAM3C or FAM3C antibody. After 24, 48 and 72 h, the viability of WSU-HN6 cells was measured by CCK-8 assay, and the activation of protein kinase B (Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24, 48 and 72 h, and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells, the mRNA and protein levels of FAM3C were significantly increased in the WSU-HN6 cells (P<0.05). The viability of WSU-HN6 cells transfected with siFAM3C was significantly inhibited at 48 h and 72 h (P<0.05). siFAM3C treatment inhibited the activation of Akt (P<0.05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt (P<0.05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt (P<0.05). CONCLUSION: FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.     

嗜水气单胞菌外膜蛋白(OMP)在乳酸乳球菌中的表达及其对BALB/c小鼠的免疫保护效果

乳酸球菌(Lactococcus lactis)是一种安全级微生物,为活载体疫苗传递抗原的理想选择.嗜水气单胞菌(A eromonas hydrophila,Ah)能引起鱼类等多种动物的败血病,因其血清型众多,寻找共同保护性抗原是进行疫苗研究的重点.为探究嗜水气单胞菌AS1.927株外膜蛋白(OMP)在乳酸乳球菌中的表达和检测其表达产物对小鼠的免疫保护效果,本研究将己克隆的ompA基因经BamH Ⅰ和Xba Ⅰ双酶切后连接载体pNZ8048,转化乳酸乳球菌NZ9000,nisin诱导表达,并进行SDS-PAGE分析鉴定.结果显示,重组基因工程菌L.lactis[pNZ-ompA]表达的融合蛋白分子量约为36.2 kD,成熟蛋白分子量约为33.7 kD.将重组基因工程菌L.lac tis [pNZ-ompA]口服免疫BALB/c小鼠(Mus musculus),用放射免疫法检测末次免疫一周后的各组小鼠肠粘膜分泌型免疫球蛋白A(sIgA)的水平以及用间接ELISA法检测小鼠血清特异性抗体IgG,结果发现,该重组基因工程菌能显著提高sIgA的分泌(P＜0.05);同时在小鼠血清中的特异性抗体含量也显著高于对照组(P＜0.05).末次免疫两周后用100 LD50的Ah AS 1.927(3.3×105 cfu/mL)腹腔注射攻击小鼠,口服重组菌对小鼠的相对免疫保护力(relative percent survival,RPS)为87.5％,表明重组基因工程菌可诱导小鼠对AhAS1.927产生一定的免疫保护作用.该结果将为开发安全有效的口服基因工程鱼用疫苗提供一定的实验基础.