Construction of a recombinant human GM-CSF/MCAF fusion protein and study on its in vitro and in vivo antitumor effects

A novel cytokine fusion protein was constructed by fusing granulocyte macrophage colony stimulat-ing factor (GM-CSF) with monocyte chemotactic activating factor (MCAF), which acts as a factor directing effector cells (monocytes) to a target site. The recombinant human GM-CSF/MCAF fusion protein could sustain the growth of GM-CSF-dependent cell line TF1 and was chemotactic for monocytes. The in vitro antitumor effect showed that rhGM-CSF/MCAF could activate monocytes to inhibit the growth of several human tumor cell lines, including a promyelocyte leukemia cell line HL-60, a lung adenocarcinoma cell line A549, a hepatoma cell line SMMC-7721 and a melanoma cell line Bowes. Furthermore, the cytotoxicity of monocytes activated by rhGM-CSF/MCAF against HL-60 and A549 was greater than that activated by GM-CSF or MCAF alone, even greater than that activated by a combina-tion of GM-CSF and MCAF, suggesting that the fusion protein has synergistic or enhanced effects. The in vivo anti-tumor effect indicated that     

Construction of a trivalent candidate Shigella vaccine strain with host-vector balanced-lethal system

A trivalent liveShigella vaccine candidate FSD01 against S.flexneri 2a, S.sonnei and S.dysenteriae I was constructed. This candidate strain was based on the S.flexneri 2a vaccine T32. By homologous recombination exchange, the chromosomalasd gene of T32 was site-specifically inactivated, resulting in the strain unable to grow normally in LB broth, while anotherasd gene of S.mutans was employed to construct an Asd⁺ complementary vector. This combination ofasd ^- host/Asd⁺ vector formed a balanced-lethal expression system in T32 strain. By use of this system, two important protective antigen genes coding for S.sonnei Form I antigen and Shiga toxin B subunit were cloned and expressed in T32, which led to the construction of trivalent candidate vaccine FSD01. Experimental results showed that this strain was genetically stable, but its recombinant plasmid was non-resistant. Moreover, it was able to effectively express trivalent antigens in one host and induce protective responses in mice against the challenges of the above threeShigella strains.     

八肽胆囊收缩素在猪,大鼠和豚鼠肠道的免疫组织化学定位和分布

实验采用ＡＢＣ免疫组织化学方法研究了八肽胆囊收缩素样免疫反应物质在猪、大鼠和豚鼠肠道的定位与分布,并用相邻切片免疫双标记法观察了它与５－羟色胺的关系,结果表明,ＣＣＫ－８－ＩＲ细胞主要位于肠腺的底部,少数位于绒毛上皮。节段性分布上,在猪可见于从十二指肠到结肠全长的粘膜,大鼠和豚鼠ＣＣＫ－８－ＩＲ细胞则见于从十二指肠至回肠的粘膜,但均以十二指肠密度最高;免疫双标记法证实,在三种动物肠道中均有ＣＣＫ＝     

类杆菌5-硝基咪唑抗性质粒pIP419的研究

5-硝基咪唑类药物广泛应用于厌氧菌感染症的临床治疗,近几年厌氧菌5-硝基咪唑抗性菌株的报道有所增加。本文介绍分离自类杆菌的编码抗5-硝基咪唑抗性质粒pIP419的研究结果。pIP419分子量为10kb,已建立了限制性内切酶物理图谱。该质粒可通过接合和转化在不同的菌种或菌株间转移。质粒上负责质粒复制和转移的基因片段已被定位和克隆。     

FXR在胃粘膜肠化生及胃癌中的表达及其意义研究

目的:研究FXR在胃炎,胃粘膜肠化生及胃癌组织中的表达,分析其在胃癌发生中的意义。方法:采用免疫组化方法检测FXR在55例胃炎组织,61例胃黏膜肠化生组织及61例胃癌组织中的表达,利用统计学方法 SPSS17.0软件分析其在三种组织中的表达变化,结合文献回顾,分析FXR在胃癌发生中的意义。结果:FXR在胃黏膜肠化生中的表达明显高于胃炎组织(P0.05),而在胃癌组织中,FXR的表达显著低于胃粘膜肠化生组织(P0.05)。结论:FXR是一个潜在的胃癌发生生物标记物,其具体机制有待于进一步探索。     

基于“心与小肠相表里”理论探讨肠道微生态与骨质疏松症的关系

近年来相关研究显示,肠道微生态在骨质疏松症的发生发展中起着重要作用。中医脏腑理论密切关注脏腑之间的生理病理关系,以中医经典《内经》“心与小肠相表里”理论为基础,探讨心、小肠、肠道微生态与骨质疏松症之间的关系。研究发现肠道微生态可能是心系疾病导致骨质疏松症的途径之一,这一发现可能为骨质疏松症的研究与防治提供一定的理论依据。     

MicroPhenoDB Associates Metagenomic Datawith Pathogenic Microbes,Microbial Core Genes,and Human Disease Phenotypes

Microbes play important roles in human health and disease. The interaction between microbes and hosts is a reciprocal relationship, which remains largely under-explored. Current computational resources lack manually and consistently curated data to connect metagenomic data to pathogenic microbes, microbial core genes, and disease phenotypes. We developed the MicroPhenoDB database by manually curating and consistently integrating microbe-disease association data. MicroPhenoDB provides 5677 non-redundant associations between 1781 microbes and 542 human disease phenotypes across more than 22 human body sites. MicroPhenoDB also provides 696,934 relationships between 27,277 unique clade-specific core genes and 685 microbes. Disease phenotypes are classified and described using the Experimental Factor Ontology (EFO). A refined score model was developed to prioritize the associations based on evidential metrics. The sequence search option in MicroPhenoDB enables rapid identification of existing pathogenic microbes in samples without running the usual metagenomic data processing and assembly. MicroPhenoDB offers data browsing, searching, and visualization through user-friendly web interfaces and web service application programming interfaces. MicroPhenoDB is the first database platform to detail the relationships between pathogenic microbes, core genes, and disease phenotypes. It will accelerate metagenomic data analysis and assist studies in decoding microbes related to human diseases. MicroPhenoDB is available through http://www.liwzlab.cn/microphenodb and http://lilab2.sysu.edu.cn/microphenodb.     

房颤射频消融术后复发风险预测评分系统的建立及评价

目的:探讨心房颤动(房颤)患者射频消融术后复发的风险因素,并依此构建个性化的风险评分系统。方法:选取2017年1~8月行射频消融术的房颤患者154例作为研究对象,依据术后3个月的随访结果将患者分为复发组及未复发组,采用单因素分析和Logistic回归分析对各风险因素进行分析,构建其评分系统,采用Hosmer-Lemeshow拟合优度检验和ROC曲线下面积评价评分系统的准确度及区分度。结果:术后随访3个月的结果显示共37例(24.03%)房颤患者出现复发,房颤类型、病程、体质量指数(BMI)、左房前后径(LAD)、左房容积(LAV)及超敏C反应蛋白(hs-CRP)水平均是房颤复发的独立风险因素(P<0.05)。构建的风险评分系统得分为0~26分,Hosmer-Lemeshow拟合优度检验:x^2=7.520,P=0.482;ROC曲线下面积为0.864(95%CI:0.837~0.891),预测评分值为15分时,约登指数最大(0.605),此时的敏感度和特异度分别为77.3%和83.2%。结论:房颤患者射频消融术后的复发率较高,依据风险因素构建的风险评分系统具有较高的预测效率和区分能力,可作为房颤患者射频消融术后复发风险评估的参考工具。     

Etk敲除及过表达细胞株构建并探讨Etk对细胞增殖的影响

目的:利用CRISPR-Cas9技术在人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVECs)中构建Etk(Epithelial and endothelial tyrosine kinase)敲除稳定细胞株以及利用慢病毒构建Etk过表达稳定细胞株,并初步探讨Etk基因对HUVECs细胞增殖的影响。方法:利用CRISPR-Cas9技术,使用在线工具设计针对Etk的sgRNA (https://chopchop.rc.fas.harvard.edu/)。将sgRNA利用连接酶整合到病毒载体中,包被病毒并感染HUVECs敲除HUVECs中的内源性Etk。利用嘌呤霉素筛选得到Etk敲除的稳定细胞株。PCR扩增Etk基因序列,将其整合到pLEX-MCS慢病毒过表达载体中构建Etk过表达重组质粒。包被病毒并感染HUVECs,在HUVECs中过表达Etk,利用嘌呤霉素筛选得到Etk过表达的稳定细胞株。利用qRT-PCR和Western-Blotting检测Etk的敲除和过表达情况。利用CCK-8盒子检测两种稳定细胞株细胞增殖情况。结果:利用CRISPR-Cas9技术有效的敲除HUVECs中内源性Etk,同对照相比,Etk的mRNA和蛋白水平显著地降低(P0.01)。同时利用慢病毒在HUVECs中过表达Etk,同对照相比,在过表达Etk稳定细胞株中Etk的mRNA和蛋白表达显著上调(P0.01)。CCK-8检测发现,Etk敲除降低细胞增殖能力;而Etk过表达增加细胞增殖能力。结论:通过CRISPR-Cas9技术成功在HUVECs中敲除Etk,利用慢病毒过表达系统成功的在HUVECs中过表达Etk,并且初步验证了Etk促进HUVECs细胞增殖。