Development of Complex Probiotics

[目的]研制益生菌复合制剂。[方法]分离、筛选纳豆芽孢杆菌XW1、乳酸菌XW2、酵母菌XW3,进行动物灌胃试验与肠道菌群检测试验。[结果]通过饲喂益生菌复合制剂,小鼠肠道厌氧菌菌群有不同程度地增多,且一些致病肠杆菌受到抑制。综合分析,得到1个益生菌复合制剂的适宜配方(108 Cfu/M l):纳豆芽孢杆菌XW1:乳酸菌XW2:酵母菌XW3为2∶1∶1。[结论]益生菌复合制剂有促进动物生长,维持肠道微生态平衡的功效。[Objective] To develop complex probiotics.[Method] Probiotic strains including bacillus natto xw1,laciobacillus xw2,microzyme xw3 were isolated.Animal feeding experiment and intestinal flora detecting experiment were carried out.[Result] After feeding by complex probiotics,the counts of anaerobic bacteria in intestine of rats significantly increased,and the counts of some pathogenic E.coli significantly declined.Througy comprehensive analysis,a appropriate scheme of complex probiotics(108 cfu/ ml)was abtained∶bacillus natto xw1∶laciobacillus xw2∶microzyme xw3 was 2∶1∶1.[Conclusion]The growth of rats and balance of bacteria in rats intestine were promoted by complex probiotics

Essence of tongue diagnosis of HE Meng-yao

岭南,指中国五岭以南的地区,相当于现在广东、广西及海南全境。因气候炎热潮湿,人群多脾胃虚弱,病多阳虚、痰湿。从晋代以后,当地医家以中原医学为基础; ,结合岭南特殊气候和人群体质特点,逐渐形成独具风格的岭南医学流派。何梦瑶作为岭南名医之一,其医术高明,医学著作颇丰,极大地促进了岭南医学的发展。; 《医碥》是其代表作之一,亦是其学术思想的精髓,为现代研究何梦瑶学术思想和岭南医家流派的主要古籍。其中所记载的舌诊方法,至今仍具有临床实用意义。Lingnan, Chinese refers to the south of the five ridges area, which is; equivalent to the current Guangdong province, Guangxi province and; Hainan province. All due to the hot and humid climate, majority of local; people are with spleen and stomach deficiency, most of the clinical; manifestation are yang deficiency and dampness. After the Jin dynasty,; based on the traditional Chinese medicine of central plains, the local; doctors gradually formed their unique medical schools, combining with; the characteristics of the local climate and peoples constitutions. As; one of the famous specialists in the south of the five ridges, HE; Meng-yao, greatly promote the development of local medical health level.; Yibian is one of his famous masterpieces, which is also the essence of; the his medicine. Records of tongue diagnosis in this book are still; practical in clinical.国家自然科学基金资助项目; 福建中医药大学校管课题; 福建省科技计划引导性项

基于微流控技术的循环肿瘤细胞分析研究进展

癌症的精准诊疗是提高癌症患者生存率和生存质量的重要手段.液体活检通过采用非侵入采样方式,获取肿瘤病人全面、准确、实时的基因组、转录组及蛋白组等生物学信息,是一种新兴的癌症诊断技术,对癌症精确诊断、个体化治疗、预后评估等方面具有重要意义.循环肿瘤细胞(CTC)是一种从实体瘤组织脱落进入外周血的肿瘤细胞,因能提供完整的细胞生物学信息,是最具应用前景的液体活检靶标.然而, CTC的数量极其稀少、异质性强、所处外周血环境复杂等特点,给CTC的富集和分析带来了巨大的技术挑战.本文将总结本课题组近年来发展的基于CTC液体活检策略,着重讨论在CTC识别、富集与单细胞分析等方面的研究进展

Promotion of proliferation of luminal B breast cancer cells by mesenchymal stem cells and its underlying molecular mechanisms

目的分析人脐带间充质干细胞(hUC-MSCs)对Luminal B型乳腺癌细胞生长增殖的影响,并初步探讨其可能的分子机理。方法绿色荧光蛋白(GFP)和荧光素酶共表达慢病毒感染人Luminal B型乳腺癌细胞BT474,并经嘌呤霉素筛选两周后,于荧光显微镜下观察GFP的表达情况,IVIS Kinetic成像系统拍照以观察和记录慢病毒感染后BT474细胞荧光素酶的表达情况;荧光显微镜下直接观察,结合MTS实验分析hUC-MSCs共培养或其浓缩上清处理对GFP和荧光素酶共表达BT474细胞生长增殖的影响;Western blot法检测hUC-MSCs浓缩上清处理对BT474细胞Akt和MAPK信号通路激活情况以及下游细胞周期调控蛋白Cyclin D1表达的影响;常规RT-PCR法检测hUC-MSCs中NRG-1、NRG-2、IGF-Ⅰ、IGF-Ⅱ和EGF等配体的表达。荧光素酶表达强度与细胞数量的相关性经由Excel软件行统计学分析,MTS实验数据则经由SPSS13.0统计软件行统计学分析。结果荧光显微镜和IVIS Kinetic成像系统的观察结果分别证实,GFP和荧光素酶经慢病毒载体系统的介导可在BT474细胞中成功地共表达,且荧光素酶的表达强度与细胞数量呈直线相关。MSCs共培养或其浓缩上清处理均可显著促进Luminal B型乳腺癌细胞BT474的生长增殖,其细胞存活比例分别为各自对照组的148.06%(P<0.005)和147.99%(P<0.001);MSCs浓缩上清处理同时激活BT474细胞内Akt和MAPK信号通路,并上调细胞周期调控蛋白Cyclin D1表达。此外,RT-PCR结果显示,hUC-MSCs中NRG-1和EGF的mRNAs水平呈高表达,而NRG-2、IGF-Ⅰ和IGF-Ⅱ等配体的mRNAs表达也可见。结论 MSCs可通过表达并分泌NRG-1等配体,从而激活Luminal B型乳腺癌细胞BT474的下游Akt和MAPK信号转导通路以上调细胞周期调控蛋白Cyclin D1的表达,进而促进其生长增殖。Objective To investigate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs) on proliferation of luminal B breast cancer cells and its underlying molecular mechanisms. Methods Human luminal B breast cancer cells BT474 were infected with GFP and luciferase co-expressing lentiviruses and then subjected for selection with Puromycin for 2 weeks. The expression of GFP and luciferase was detected by fluorescent microscopy and IVIS Kinetic image system, respectively. The effect of coculture or treatment with conditioned medium of hUCMSCs on proliferation of BT474 was analyzed with MTS assay. Western blot was carried out to detect the effect of treatment with conditioned medium of hUC-MSCs on the activation of both Akt and MAPK signalings in BT474, as well as the expression of downstream cell cycle regulator Cyclin D1. Regular RT-PCR was applied to analyze the mRNAs expression of ligands such as NRG-1, NRG-2, IGF-Ⅰ, IGF-Ⅱ, and EGF in hUC-MSCs. The correlation between relative luciferase activity and cell number was analyzed with Excel software, while MTS assay data was statistically analyzed with SPSS 13.0 software. Results The co-expression of GFP and luciferase in BT474 via lentiviral expression system was visualized by fluorescent microscopy and IVIS Kinetic image system. The linear correlation between relative luciferase activity and cell number was determined by curve fitting analysis. Coculture or treatment with conditioned medium of hUC-MSC significantly promoted the proliferation of BT474, with survival rates being 148.06 %(P < 0.005)and 147.99 %(P < 0.001)of control, respectively. In addition, treatment with conditioned medium of hUC-MSC was shown to induce activation of both Akt and MAPK signalings, which further upregulated the expression of Cyclin D1. Moreover, high mRNAs expression levels of both NRG-1 and EGF, as well as moderate mRNAs expression levels NRG-2, IGF-Ⅰ, and IGF-Ⅱ were showed by RT-PCR. Conclusion Our results here demonstrated that MSCs may promote the proliferation of luminal B breast cancer cells through paracrine of ligands such as NRG-1, which in turn results in the activation of both Akt and MAPK signalings and upregulation of the expression of Cyclin D1.国家自然科学基金面上项目(81272922);; 福建省自然科学基金面上项目(2016J01577);; 福州总医院院内课题国际合作研究专项(2015G01

Improvement on Determination of Lead in Blood by Graphite Furnace Atomic Absorption Spectrometry

[目的]改进石墨炉检测低浓度血铅国标方法,对测定结果进行不确定度评定。[方法]应用0.20%硝酸和0.10%曲拉通X-100混合溶液作为稀释剂,0.10%硝酸钯和0.06%硝酸镁作为基体改进剂,参考仪器推荐测定条件,应用AA-900T石墨炉原子吸收分光光度计测定血中铅。对所建立的方法进行检出限及定量下限等方法学验证,并应用国家标准物质对准确度进行评定。[结果]结果显示:检出限为0.84μg/l;定量下限为2.80μg/l;线性范围为2.80~200.00μg/l;血铅浓度为100.00μg/l时相对标准偏差为1.24%;对国家标准物质进行测定,检测结果为112.10μg/l,落在其标准值及不确定度范围之内[(120.00±15.00)μg/l]。[结论]所建立的血铅检测方法各项确认参数符合要求,方法的不确定性在容许范围内。[Objective]To improve a method on determination of lead in blood by graphite furnace atomic absorption spectrometry and to estimate the uncertainty of the application.[Methods]Mixed solution containing 0.20% nitric acid and 0.10% Triton X-100 was used as diluent,and mixed solution containing 0.10% palladium nitrate and 0.06% magnesium nitrate was used as matrix modifier.Referring to the recommended instrument measuring conditions,an AA-900T graphite furnace atomic absorption spectrophotometer was applied to determine lead in blood.The limits of detection and the lower limit of quantification of the proposed revised measurement were estimated.The accuracy were determined by national standard reference material.[Results] The detection limit was 0.84μg/L and the quantification lower limit was 2.80μg/L.The linear range was 2.80-200.00μg/L,and the relative standard deviation was 1.24% for blood lead at 100.0μg/L.The concentration of the national standard reference material was 112.10μg/L and within the required uncertainty range of(120.00±15.00) μg/L.[Conclusion]All parameters of the proposed method meet the requirement of the national clinic laboratory technique guideline.The estimated uncertainty of the method falls within the acceptable range.北京市自然科学基金资助项目(编号:2122060

Process technology on recovery of sulphur from copper-bearing gold concentrate by using kerosene method

某含铜金精矿尾矿浸铜渣中含有质量分数约15%的单质硫,严重影响后续提金过程的氰化效果。实验进行了以煤油为溶剂的提硫过程研究,考察溶硫时间、温度、液固质量比等因素。实验表明:采用煤油二段浸取法,当工艺条件为液固质量比2—3,温度140℃,加热40MIn,单质硫的回收率为98.4%,纯度达99%以上。对硫化矿处理过程尾矿中硫的回收,采用煤油为溶媒,是一种有效、环保的清洁生产过程,可以提高贵金属及伴生硫的综合回收率,易于工业应用。In the residues of copper-bearing gold concentrate there is mass fraction 15% sulfur which influences the effect of latish cyanogenation.The experiment was carried out with kerosene as solvent in sulfur removal process and the effect of leaching temperature and time,the mass ratio of kerosene to residue (liquid to solid) on the experiment was investigated.The results show that when the mass ratio of kerosene to residue is 2-3,heating temperature is 140 ℃,heating time is 40 min,the recovery efficiency of sulfur can be more than 98.4%,the purity of sulfur can be more than 99%.The new hydrometallurgical method which uses kerosene as solvent is an effective and clean process.It can improve comprehensive recovery efficiency of precious metals and sulfur,and is easy for industrialization without pollution

铅胁迫对双孢蘑菇积累铅及抗氧化酶活性的影响

铅（Pb）是环境中最普遍的重金属污染物之一，不仅直接毒害土壤生物，破坏生态结构，还可以通过食物链迁移转化，危害人体健康，因此Pb污染的治理迫在眉睫。大型真菌具有强的金属富集能力及较高的相对年生物量，因此在对Pb污染的修复中有一定优势。本文以双孢蘑菇菌丝为研究对象，采用马铃薯葡萄糖琼脂培养的方法，研究不同浓度Pb胁迫对双孢蘑菇菌丝体的生长状况、形态以及机体内抗氧化酶系统的影响。结果表明，在实验浓度下Pb对双孢蘑菇菌丝体的生长率无显著性影响，且菌丝体中Pb含量随着培养基中Pb浓度的增加而升高，达到10.40 μg/g。扫描电镜和透射电镜结果显示，Pb胁迫下菌丝体的细胞结构受到一定破坏，细胞膜透性增加。X射线能谱分析观察得到细胞中的黑色颗粒为铅，表明Pb在细胞内的富集。菌丝体中丙二醛含量随Pb浓度的增加而增加，超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）等抗氧化酶活性随着Pb浓度的增加有不同程度的反应。在酶活力达到最大时，相比对照组分别增加了56.45%、61.36%、13.08%和10.59%，说明双孢蘑菇通过调节自身抗氧化酶系统适应Pb胁迫环境，据此在Pb污染的生物修复中具有应用潜力。福建省教育厅中青年教师教育科研项目（JAT170364）;;\n国家自然科学基金项目（31530008和318704832）资

Outline of outcomes of the “Declaration of International Japan-Studies” 21st Century COE Research Program

The basic concept behind the formation of a center for the “Declaration of International Japan-Studies” is the development of Japanese studies as cross-cultural research. Building on this concept, the program has produced the following four main outcomes.1) The first outcome concerns research approaches and methodologies, and is apparent in the adoption of a cross-cultural approach to Japanese studies through the incorporation of the perspectives of foreign researchers on Japanese studies (i.e., perspectives of “the other”). The construction of international Japanese studies is a meta-scientific venture that entails exploration of the conditions for the establishment of academic dialogue at home and abroad through dialogue between Japanese and non-Japanese researchers based on open approaches to research. This has yielded a model for international and academic collaborative research that overthrows the archaic modi operandi of the humanities in Japan—the closed, inward-looking approach and poor awareness of method—by founding a new academic discipline backed by the clear methodology of international Japanese studies.2) The second outcome has been the rediscovery of Japanese culture from a cross-cultural perspective and the opening of a new realm in Japanese cultural studies. Abroad, “Japanese studies” has consisted of research on Japan as “another culture,” and so has always incorporated a comparative element. This comparative perspective, particularly when it has taken the form of comparison between Japan and China, Japan and Germany, and Japan and France, has enabled the rediscovery of new and unconventional aspects of Japanese culture through the reinterpretation of Japanese culture itself. The location of nō, designated by UNESCO one of the “Masterpieces of the Oral and Intangible Heritage of Humanity,” within the framework of international Japanese studies and its reinterpretation from a comparative perspective has opened up new realms in nō studies, while the shift in interest to the southern boundary region of the Ryukyus and Okinawa has demonstrated that Japanese culture, rather than being a single uniform and homogenous culture, in fact consists of “many cultures” having multiple cross-cultural sources and a multilayered structure that extends beyond Japan\u27s borders to China, Korea, and other parts of Asia.3) The third outcome has been the promotion of joint research activities on a global scale through the sharing of research data in real time and effective use of networks with researchers and research institutes in Japan and overseas. To assist in this process, content of value to Japanese studies has been converted to electronic format to create an electronic library system accessible to researchers from around the world.4) Finally, the fourth outcome has been the development of researchers in the field of Japanese studies who are equipped to communicate their findings to a wider domestic and international audience, achieved by applying the fruits of the “Declaration of International Japan-Studies” COE Program to postgraduate education at Hosei University\u27s Institute of International-Japan Studies

Preparation and biological safety of basic fibroblast growth factor/double-layered collagen composite

背景:胶原特殊的分子结构和生物活性有利于多种细胞黏附、增殖和分化,并可降解为新生组织提供足够空间。目的:制备一种复合负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子双层胶原基复合材料,并评价其生物安全性。方法:制备交联风干胶原膜和交联冻干胶原膜。将壳聚糖-肝素纳米粒滴于交联冻干胶原膜上,再将湿态交联风干胶原膜置于复合纳米粒子的交联冻干胶原膜上风干,即碱性成纤维细胞生长因子/双层胶原基复合材料。采用急性全身毒性试验、溶血试验、热原试验和细胞毒性试验评价其生物安全性。结果与结论:碱性成纤维细胞生长因子/双层胶原基复合材料为双层结构,一侧表面致密,另一侧疏松多孔。在其中间负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子呈不规则球形分布于胶原膜内侧面;急性全身毒性试验、热原试验、溶血试验均为阴性,细胞毒性为0级。说明碱性成纤维细胞生长因子/双层胶原基复合材料具有良好的生物安全性,对机体无毒,符合ISO10993-1评价标准。BACKGROUND: Collagen is a common used scaffold in tissue engineering, its specific molecular structure and biological activity are conducive to a variety of cell adhesion, proliferation and differentiation, and can speed up wound healing and degradation to provide sufficient space for new tissues.OBJECTIVE: To prepare a kind of inhomogeneous double layered collagen composite incorporated with basic fibroblast growth factor (bFGF) loaded chitosan-heparin (CS-Hep) nanoparticles, and to evaluate its biological safety.METHODS: The air-dried dense layer collagen membranes and freeze-dried loose layer collagen membrane were prepared respectively and then cross-linked by D-ribose (termed as CAM and CFM respectively).CS-Hep nanoparticles were prepared and then dropped on the surface of CFM.The wet CAM was laid on the surface of CFM with nanoparticles, and then air dried completely at 4 ℃.The obtained material was termed as bDM.Its physical and chemical properties were investigated.In addition, its bio-safety was also examined through acute systemic toxicity, pyrogen, hemolysis and cellular toxicity tests.RESULTS AND CONCLUSION: The bFGF/double-layered collagen composite was made up of double layers with one dense layer (CAM) and another loose layer (CFM).As part of bDM, CS-Hep nanopartiles with irregular global shape were aligned in the scope of D-period structure of the inner surface of CFM; the bFGF/double-layered collagen composite had no systemic acute toxicity, no cytotoxicity and pyrogen reactions as well as no hemolytic effect.The bFGF/double-layered collagen composite is prepared and found to have good biocompatibility and safety.Moreover, this material conforms to the ISO 10993-1, and can be used as a basic scaffold material in tissue engineering天津市科技支撑计划重点项目(08ZCKFSF02100);博士点基金项目(20101106110042)---

50味中药及其复方与抗生素对河流弧菌的体外抑菌作用

用牛津杯法选择大黄、黄柏、黄连等50种单味中药进行河流弧菌、金黄色葡萄球菌体外抑菌试验,并以抗菌活性较好的药物组成二联、三联复方,药物质量浓度分别为60和240 mg·mL-1,进行中药复方抗菌活性研究,同时用抗生素进行河流弧菌药敏试验。目的是筛选出对河流弧菌敏感的中草药、抗生素及其复方。结果表明,黄连、诃子等单方对河流弧菌抑菌作用明显,为极敏感;在60 mg·mL-1低药物浓度时,黄柏、黄连组成的复方对河流弧菌抑菌作用为极敏感;在240 mg·mL-1药物浓度时,黄柏、黄连等组成的9个复方中药对河流弧菌抑菌作用属极敏感,黄连、板蓝根、黄柏等4个复方中药对河流弧菌的抑菌作用明显,为极敏感。河流弧菌对恩诺沙星、盐酸多西环素、氟苯尼考、土霉素及甲氧苄啶/磺胺甲噁唑5种药物高度敏感。部分中药对河流弧菌抑制作用不明显,黄连、板蓝根、黄柏等4个三联复方中药药效最强。可组成多种三联复方中药或中西药联用复方,均有良好的抑制作用。2018年开放课题基金项目(LYC2018RS04,闽海鸥[2018]31号)福建省自然基金项目(2016J01167,2018J01455)国家自然科学基金(31702384,31502194)福建省科技重大专项(2016NZ0001-3)2017年集美大学研究生教育教学改革研究项目(集大研No.[2017]18号)教育部鳗鱼工程研究中心开放基金项目(RE201704