细胞三维动态培养微器件的设计与制作

细胞培养是进行细胞研究的基础,为了在细胞体外培养时提供一种近似于体内的微环境,设计了一种可供细胞三维动态培养的微器件。首先设计了用于输运流体的微通道网络,培养池对称布置于微通道网络中,通过一系列\"多进多出\"型微通道分别与进样口和出样口相连。利用Comsol软件中的层流物理场和多孔介质物理场耦合对培养池内的流场进行仿真,通过比较流场的均一性和稳定性优化微通道网络结构。然后,采用静电直写技术在培养池内集成聚己内酯(PCL)三维支架,构建细胞三维培养空间。最后,封合微器件,检测微器件培养池内的流体流动情况,并进行细胞实验。实验结果表明,\"2×2\"型微器件培养池内的流体稳定性和均一性较好;PCL三维支架的纤维间距400μm,纤维直径80μm,孔隙率64%,细胞存活率达到90%以上。该细胞三维动态培养微器件更好地模拟了生物体内细胞生存所需的微环境,培养池内的细胞生长良好,满足设计要求。国家自然科学基金资助项目(No.51475079,No.51375076

“动物酵素营养液”对母猪便秘的影响

从200头母猪中选出同一时期怀孕的母猪40头做两次试验,以验证“动物酵素营养液“对母猪便秘的影响。结果显示,A组试验,试验组便秘猪痊愈,治愈时间短,且没发现复发情况。b组试验,对照组有便秘情况发生,腹泻发病率达14.46%,仔猪死淘率达13.25%,试验组无便秘发生,腹泻发病率仅为3.125%,仔猪死淘率仅为1.56%。黑龙江省教育厅自然科学研究项目:酵素在黑龙江省养猪生产中的应用研究。项目编号为1253515

动物酵素营养液对保育猪呼吸道疾病影响的研究

本试验选择28日龄的仔猪,喂养动物酵素营养液至70日龄,结果表明:酵素组的仔猪中,康复率为75%,28.6%完全康复,57.1%转为轻微患病,14.3%处于严重状态,在对照组中只有40%完全康复,严重时有1头死亡,4头转为轻微患病,说明动物酵素营养液对仔猪呼吸道疾病有较好的促进康复的效果

动物酵素营养液对繁殖母猪生产性能的研究

为验证“动物酵素营养液“对母猪生产性能的影响,选120头母猪进行饲养试验。结果表明,“动物酵素营养液“能够提高母猪群产仔率、成活率,平均窝产仔最多12.5头,成活率最高可达98%~99%,仔猪出生重为1.60千克/头,断奶后仔猪体重为11.2千克/头。猪无腹泻现象,长势快,皮毛光滑,整齐度提高,母猪妊娠期正常,产程时间少,无三炎,母猪健康,可在产后3~4天开始发情,配种率与受胎率均为100%,每年可产2.4窝,每头母猪与对照组的经济差额为4 520元

Screening of adjuvant enhancing cellular immune response induced by ESAT6-CFP10 fusion protein in mice

通讯作者：叶祥忠，E-mail：[email protected][中文文摘] 目的 筛选能增强特异性抗原早期分泌抗原靶6蛋白(Early secretory antigenic target6,ESAT6)-培养滤出液蛋白-10(Culture filtrate protein10,CFP-10)融合蛋白(E1C0)诱导小鼠细胞免疫应答的佐剂,建立基于细胞免疫应答的小鼠模型,以评价基于体外干扰素γ释放分析(IFNγrelease assay,IGRA)结核诊断方法中特异性刺激抗原E1C0的活性。方法 建立小鼠IFNγ双抗体夹心SABC-ELISA检测系统,并验证系统的线性、灵敏度、重复性和特异性。将BALB/c小鼠随机分为7组:E1C0+单磷酸类脂A(Monophosphoryl lipid A,MPL)+双十八烷基二甲基溴化铵(Dimethyl dioctadecylammonium bromide,DDA)组、E1C0+DDA组、E1C0+MPL组、E1C0+弗氏不完全佐剂(IFA)组、E1C0组、生理盐水组和MPL+DDA联合组,每组6只,经小鼠后肢内侧皮下免疫3次,间隔2周,免疫剂量为:E1C0100滋g/只,MPL25μg/只,DDA250μg/只,IFA100滋l/只。末次免疫4周后处死小鼠,无菌取脾,分离脾淋巴细胞,加入E1C0进行培养,MTT法检测特异性淋巴细胞增殖反应,ELISA法检测培养上清中IFNγ水平。采用筛选出的最佳佐剂与抗原组合免疫3批BALB/c小鼠,进行IFNγ诱生测定。结果 检测系统的线性范围为:40～2560pg/ml(R>0.98);灵敏度为40pg/ml;变异系数(CV)0.05)。结论 E1C0与MPL和DDA联合免疫所诱导的小鼠Th1型细胞免疫应答最强,成功建立了用于评价刺激抗原E1C0活性的小鼠模型。[英文文摘]Objective To screen the adjuvant enhancing the cellular immune response induced by early secretory antigenic target 6（ESAT6）-culture filtrate protein-10（CFP10）in mice, and establish an animal model based on cellular immunγe response for evaluation of activity of specific stimulating antigen E1C0 in IFNγ release assay（IGRA）for diagnosis of tuberculosis（TB）. Methods mDouble antibody sandwich SABC-ELISA system for mouse IFNγ was developed and verified for linearity, sensitivity, reproducibility and specificity. BALB/c mice were randomly divided into seven groups, 6 for each, and immunized s.c. with E1C0 + monophosphoryl lipid A（MPL）+ dimethyl dioctadecylammonium bromide（DDA）, E1C0 + DDA, E1C0 + MPL, E1C0 + IFA, E1C0, physiological saline and MPL + DDA for 3 times, respectively, each at an interval of 2 weeks. The dosages of E1C0, MPL, DDA and IFA for immunization were 100 μg, 25μg, 250μg and 100 μl, respectively. The mice were killed 4 weeks after the last immunization, and their spleens were collected aseptically, from which splenic lymphocytes were isolated, cultured with E1C0, then determined for proliferation level by MTT method, and for IFNlevel in culture supernatant by ELISA. Three batches of BALB/c mice were immunized with the screened adjuvant combined with antigen, and determined for IFNγ induced. Results The linear range, sensitivity and CV value of developed SABC-ELISA system were 40 ~ 2 560 pg / ml（R > 0. 98）, 40 μg/ml and less than 15%respectively, by which all the detection results of IFN酌in rat, guinea pig and rabbit sera were negative. The stimulating indexox（SIs） of specific lymphocyte proliferation in E1C0 + MPL + DDA, E1C0 + IFA and E1C0 + DDA groups were significantly higher than those in physiological saline group （P < 0. 01）. The IFN酌level secreted by lymphocytes in E1C0 + MPL + DDA group after stimulation with E1C0 in vitro was significantly higher than those in other groups （P < 0. 001）. No significant differences were observed in IFNγ levels induced in 3 batches of mice in E1C0 + MPL + DDA group（P > 0. 05）. Conclusion The immunization with E1C0 in a combination with MPL and DDA elicited a strong Th1 cellular immune response in mice. Mouse model for evaluation of activity of E1C0 antigen was successfully established

Experimental study on reperfusion of intraocular lens

作者简介: 祁明信, 男, 1945 年7 月 出生, 教授、主任医师、博士研究生 导师, 主要从事白内障的基础与临 床研究。联系电话: 0591-83570887; E-mail:qihuang@netease. com 通讯作者: 黄秀榕,E-mail:[email protected][中文文摘]目的开展晶状体再灌注的离体和动物实验研究,并对再灌注人工晶状体技术进行评价。方法采用新鲜离体幼兔眼、离体猪眼、新西兰白兔眼,应用自行研制的人工晶状体材料,进行以下实验:(1)体外固化实验;(2)晶状体前囊膜微型撕囊及其稳固性实验;(3)经微型前囊膜开口超声乳化吸出晶状体内容物实验;(4)活的新西兰白兔眼内人工晶状体再灌注实验。结果(1)按硅酮聚合物与固化剂50:1的比例可获得柔软、弹性好、固化时间短(完全固化时间为60min)的注入材料;(2)晶状体前囊膜1.8～2.0mm的连续环形撕囊口具有较好的稳定性,可经该微型开口吸出晶状体内容物并灌注材料;(3)超声能量18%、流量25mL·min-1、负压120mmHg(1kPa=7.5mmHg)为晶状体内容物经微型前囊膜开口吸出的最佳条件;(4)注入灌注材料后可形成由晶状体囊膜包裹的、置换原晶状体皮质和核的、新的再灌注人工晶状体。结论采用再灌注人工晶状体的方法可进行新型人工晶状体再灌注,可为治疗白内障和老视提供参考。[英文文摘]Objective To carry out the experimental study on reperfusion of intraocular lens(IOLs) in vitro or in animal,and to assess the technique of IOLs reperfusion.Methods The following experiments were performed by using self-developed materials in fresh rabbit eyes and pig eyes in vitro,as well as in eyes of alive New-Zea-land rabbits:(1)Solidification study of self-developed material in vitro;(2)Continuous circular capsulorhexis(CCC) in anterior capsule of lens and its stability;(3)Draw of lens contents via phaco through mini-CCC;(4)IOLs ref illing in the eyes of alive New-Zea land rabbits. Results(1) Thematerialwhich was soft, springy and short-term solidification(full solidification time was 60 minutes) were obtained in certain proportion of geland solidified agent(50:1) in vitro; ( 2)The CCC in anterior capsule of lens with 1.8-2.0 mm diameter had very good stability. The lens contents were drawn and the materialwere refilled through themini-CCC; (3) The best conditions of drawing out lens contents through m ini-CCC were phaco energy 18% , flow 25 mL·min- 1, and negative pressure 120 mmH g (1kPa=7.5 mmHg);(4) The new refilled IOLs, which were wrapped by capsule of lens and were replaced original cortex and nucleus of lens, were obtained after thematerial refillied. Conc lusion. New IOLs are refilled through this method, which can prov ide reference for the treatment of cataract and presbyopia.福建省科技三项费用;教育厅重点资助项目基金资助(编号:K98041

城市生活垃圾焚烧体系化学热力学平衡分析

采用化学热力学平衡分析方法，分析城市生活垃圾焚烧体系污染物的排放特性。主要计算了聚氯乙烯(PVC)及典型城市垃圾焚烧过程氯元素的化学平衡组成及其浓度，同时研究了脱氯剂对PVC及垃圾中氯转化的影响。计算结果表明，PVC热态反应的主要产物是HCl气体，在加入脱氯剂后温度低于600℃时，脱氯剂对HCl的脱除作用很明显；城市生活垃圾热解和燃烧过程中在温度超过600℃，垃圾中的有机氯和无机氯都将转化为HCl气体，而此时脱氯剂失去效果。由计算结果得到控制HCl生成，消除二噁英污染物的生成反应工艺条件