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中国生物工程杂志 2024年第5期44卷 40-55页

作者：王瑶姜志洋蔡军雨樊榕榕贾姗姗冯健男石艳春王晶郑源强内蒙古医科大学内蒙古自治区分子生物学重点实验室呼和浩特010058 军事科学院军事医学研究院毒物药物研究所国家安全特需药品全国重点实验室北京100850

目的:利用CRISPR/Cas9技术构建POLQ基因敲除293T细胞系,并探讨该敲除细胞系在外源基因定点整合方面的应用。方法:构建靶向POLQ基因的pX330-sgRNA打靶载体,通过T7EI酶切验证,选取打靶效率最高的载体,转染293T细胞;利用流式细胞分选术、... 详细信息

目的:利用CRISPR/Cas9技术构建POLQ基因敲除293T细胞系,并探讨该敲除细胞系在外源基因定点整合方面的应用。方法:构建靶向POLQ基因的pX330-sgRNA打靶载体,通过T7EI酶切验证,选取打靶效率最高的载体,转染293T细胞;利用流式细胞分选术、稳定单细胞克隆培养、TA克隆测序及蛋白质印迹等方法验证POLQ敲除情况;利用靶向AAVS1和FBL位点的含GFP/mCherry报告基因同源打靶载体,评估敲除的293T POLQ-/-细胞系在外源基因定点整合效率的变化,并通过测序方式验证供体报告载体是否定点整合到基因组中;最后分析POLQ敲除对293T细胞增殖和存活率的影响。结果:成功在293T细胞中稳定敲除POLQ基因;通过报告基因实验验证,敲除细胞系中外源基因定点整合效率平均提高2~7倍,且敲除POLQ基因对293T细胞系增殖和存活率没有明显影响。结论:成功利用CRISPR/Cas9技术构建具有高效定点整合能力的POLQ基因稳定敲除293T细胞系,为进一步利用该细胞系高效制备稳定整合外源基因(包括抗体基因)工程化细胞系奠定基础。

关键词： CRISPR/Cas9 POLQ 293T细胞基因敲除定点整合

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维普期刊数据库

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Attenuation mechanism of Brucella melitensis M5-10,implications for vaccine development and differential diagnosis

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Frontiers of Agricultural Science and Engineering 2014年第4期1卷 331-340页

作者： yuanqiang zheng Yanchun SHI Chang AN Ruisheng LI Dongjun LIU Yuehua KE Kairong MAO Mingjuan YANG Zeliang CHEN Shorgan BOU National Research Center for Animal Transgenic Biotechnology Inner Mongolia UniversityHohhot 010021China Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical UniversityHohhot 010058China Animal Laboratory Center 302 Hospital of PLABeijing 100039China Institute of Disease Control and Prevention Academy of Military Medical SciencesBeijing 100071China China Institute of Veterinary Drug Control Center for Veterinary Drug EvaluationMinistry of AgricultureBeijing 100081China College of Medicine Shihezi UniversityShihezi 832003China

Brucellosis is a worldwide *** is the most efficient means to prevent and control *** current licensed attenuated vaccines for animal use were developed by sequential passage in nonnatural hosts that decreased virulence in its original *** attenuation mechanism of these strains remains largely *** the present study,we sequenced the genome of Brucella melitensis vaccine strain *** analysis showed that a large number of genetic changes occurred in the vaccine strains.A total of 2854 genetic polymorphic sites,including 2548 SNP,241 INDEL and 65 MNV were *** the 2074 SNPs in coding regions,1310(63.2%)were non-synonymous *** number,percent and N/S ratios were disproportionally distributed among the cog *** polymorphic sites were identified in genes of the virB operon,flagella synthesis,and virulence regulating *** data indicate that changes in some cog categories and virulence genes might result in the *** attenuation mechanisms also have implications for screening and development of new vaccine *** genetic changes in the genome represent candidate sites for differential diagnosis between these vaccine strains and other virulence *** analysis of virulence genes showed that expression of dnaK,vjbR were reduced in M5-10 strain when compared with that in 16M.A duplex PCR targeting virB6 and dnaK was successfully used to differentiate between M5-10 and the virulent 16M *** genome re-sequencing technique represents a strong strategy not only for evaluation of vaccines,but also for development of new vaccines.

关键词： Brucella live attenuated vaccine attenuation differential diagnosis

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维普期刊数据库

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Update on laboratory diagnosis of Brucellosis

Update on laboratory diagnosis of Brucellosis

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第十二届全国免疫学学术大会

作者： XU Sen zheng yuanqiang SHI Yanchun Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical University

The persistent worldwide prevalence of Brucellosis causes serious public health concerns and economic loss to *** is a multi-system disease with significant variability and the patient’s symptoms are similar to other infectious *** addition,images may be misleading to the ***,laboratory tests are needed for the diagnosis and treatment of *** separation of the pathogen is the gold standard for diagnosis of Brucellosis,but there are some problems such as long culture cycle and low detection rate,requiring highly skilled operator and special level three biological safety *** detection including PCR and RT-PCR requires more professional operator,technologies and *** the two methods are potential biological risks and not suitable for "highthroughput" *** the establishment of test tube agglutination reaction by Wright and Smith,the serological test technology has been rapidly developed and *** there are some false positive and false negative,it has many advantages including simple operation,stable reagent,low biological risk and a uniform set of *** is now widely used in medical institutions and veterinary *** now,there are some methods/techniques such as Coombs’ test,complement fixation test,ELISA,fluorescence polarization assay and so *** search for reliable,discriminatory diagnostic and prognostic markers,especially for monitoring disease evolution,are ongoing.

关键词： Brucellosis laboratory diagnosis serological test culture

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Progress in cervical cancer vaccine research

Progress in cervical cancer vaccine research

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第十三届全国免疫学学术大会

作者： Yujuan Huo yuanqiang zheng Yanchun Shi Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical University

Cervical cancer is the fourth most common female malignant tumor, and its mortality ranks second among female cancers in *** has been confirmed that human papillomavirus(HPV) plays an important role in the pathogenesis of cervical ***-term infection of high-risk HPV is the primary factor in inducing cervical ***, prevention and treatment of HPV infection is particularly *** launch of HPV prophylactic vaccines makes cervical cancer the only cancer that can be prevented early and is expected to be ***-risk human papillomavirus(HPV) infection is closely related to the occurrence and development of cervical ***, HPV preventive vaccine research is mainly aimed at high-risk HPV(HR-HPV).Vaccines currently used for cervical HPV infection are mainly divided into preventive vaccines and therapeutic *** are three prophylactic HPV vaccines approved worldwide, four-valent HPV(HPV6/11/16/18) vaccine Gardasil, bivalent HPV(HPV16/18) vaccine Cervarix and nine-priced HPV(HPV6/11/16/18/31/33/45/52/58) Vaccine Gardasil 9 has been approved for marketing and for clinical *** preventive vaccines are ineffective against people infected with HPV, the development of HPV therapeutic vaccines has become a hot topic in the field of related diseases in recent *** main types of HPV therapeutic vaccines include: HPV vector vaccines, peptide vaccines/protein vaccines, genetic vaccines, cell vaccines, and the *** HPV vaccine is still in the research stage, and the immunogenicity and safety still need to be *** article reviews the recent advances in the prevention and treatment of cervical cancer by prophylactic and therapeutic HPV vaccines.

关键词： Cervical cancer Human papillomavirus Preventive vaccine Therapeutic vaccine

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Evaluation of a recombinant bacillus calmette-guérin vaccine expressing P39-L7/L12 of Brucella melitensis:An immunization strategy against brucellosis in BALB/c mice

Evaluation of a recombinant bacillus calmette-guérin vaccin...

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第十四届全国免疫学学术大会

作者： Yanchun Shi yuanqiang zheng Yajing Chen Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical University

Brucellosis is a chronic infectious disease caused by bacteria from the genus *** has a serious global economic impact,and poses a threat to public ***,vaccines should exhibit a safe profile as well as enhanced protective ***,there are no licensed vaccines available for humans against *** this study,for the first time,a recombinant BCG(rBCG) vaccine expressing the P39-L7/1 2 fusion protein of Brucella melitensis against brucellosis was *** rBCG vaccine elicited strong antigen-specific humoral and T cell-mediated immunity in *** rBCG vaccine also induced a significant level of protection following challenge with a virulent strain of *** M28 in BALB/c ***,the protection level induced by the rBCG vaccine was sig nificantly higher than that induced by ***,these results suggested that rBCG vaccine acted as a suitable vaccine candidate and a new strategy against human brucellosis.d as a suitable vaccine candidate and a new strategy against human brucellosis.

关键词： Recombinant BCG Vaccine P39-L7/12 Brucellosis

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Macrophage polarization and the mechanism in anti-infection immunity

Macrophage polarization and the mechanism in anti-infection ...

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第十二届全国免疫学学术大会

作者： LIANG Zhina zheng yuanqiang SHI Yanchun Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical University

Macrophage polarization reflects the state of the immune environment and changes in the ***,the fact that macrophage polarization plays a major role in inflammatory response has reached a *** to flexible phenotype traits and feature-rich,mature macrophages can appear phenotype,function and morphological differentiation induced diverse factors,which is the polarization of *** are divided into classically activated macrophages(CAM or M1) and alternatively activated macrophages(AAM or M2) two categories.M1 not only engulf and destroy foreign pathogens but also activate T-cells of the adaptive immune response and advance inflammatory ***,M2 can promote tissue repair and wound healing,in addition the suppression of tumor growth,metastasis,and tumor immune also can be *** these two kinds of polarization states are *** plays an important part in the prevention to study macrophage polarization signal pathways for intervention on various inflammatory *** early TB infection and active TB patients,Lung macrophages are the type of M1,which means M1 macrophages controlling TB infection and ***,in the course of chronic tuberculosis,infiltrating macrophages tend to type M2 that can weaken the anti-TB infected t-cell immunity and promote chronic infection of tuberculosis by anti-inflammatory reactions and antigen-presenting *** of macrophages have some effect on inflammatory *** other words,subtypes of macrophages dominate inflammatory responses by controlling their changes,so that can effectively remove pathogens and validly control the excessive inflammatory response to the body’s *** into role of macrophages in immunity to infection and mechanism,is propitious to find such a new direction for the treatment and prevention of diseases.

关键词： Macrophage polarization anti-infection anti-inflammatory

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Tumor immunotherapy in the treatment of NSCLC

Tumor immunotherapy in the treatment of NSCLC

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第十二届全国免疫学学术大会

作者： SUN Xiaoying zheng yuanqiang CUI zhengrong SHI Yanchun Inner Mongolia Key Laboratory of Molecular Biology Inner Mongolia Medical University Pharmaceutics Division College of Pharmacythe University of Texas at Austin

Lung cancer is the leading cause of cancer mortality worldwide,accounting for 1.3 million deaths each ***-small cell lung cancer(NSCLC) accounts for approximately 80-85%of all lung cancers and is usually diagnosed at an advanced stage with poor *** chemoradiotherapy improves overall survival as compared to sequential chemotherapy and radiation therapy,but is associated with higher rates of *** has shown promising results in various types of ***,immune-checkpoint inhibitor(ICPI) drugs,which include antibodies against Cytotoxic T-Lymphocyte Antigen-4(CTLA-4),programmed cell death protein 1(PD-1) and programmed cell death protein ligand 1(PD-L1),have been shown to induce durable complete responses in a proportion of patients with particular efficacy demonstrated in both the first line and refractory setting in advanced *** inhibitors are the most striking innovation in the clinical development of immunotherapy.T-cell checkpoint inhibitors that target the PD-1 and PD-L1 are changing the current strategy in the treatment of advanced NSCLC without driver gene mutations such as anti-PD-1/PD-L1,pembrolizumab,are a novel class of inhibitors that function as a tumor suppressing factor via modulation of immune cell-tumor cell interaction.

关键词： NSCLC Tumor immunotherapy Immune check point Inhibitors PD-1/PD-L1

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Brucellosis In Chinese and Mongolian Medicine

Brucellosis In Chinese and Mongolian Medicine

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第十三届全国免疫学学术大会

作者： Yumei Zhou Yajing Chen yuanqiang zheng Yanchun Shi Inner Mongolia Key Lab of Molecular Biology School of Basic Medical Sciences Inner Mongolia Medical University

Brucellosis is the world’s most widespread zoonosis, but also ranks as one of the seven most neglected diseases, according to the World Health ***, it is recognized as the world’s most common laboratory-acquired *** are a reported 500,000 incident cases of human brucellosis per ***, true incidence is estimated to be 5,000,000 to 12,500,000 cases *** diagnosed, focus is directed at treating individual patients with antibiotic regimes, yet the drug resistance is common in brucellosis *** Medicine(CM) has been used for several thousand *** Chinese herbal medicine(CHM) has a long-history for treatment of various human diseases including tumors, infection, autoimmune diseases in Asian countries, especially in China, Japan, Korea and *** and Mongolian medicine had achieved great improvements in curing acute and chronic brucellosis patients.I will summarized the immunological mechanism and from Chinese medicine perspective, provide a theoretical basis for the treatment and the development of vaccines of brucellosis.

关键词： Brucellosis Chinese and Mongolian medicine treatment vaccines

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Statistical study of magnetic holes in the upstream region of Mercury’s bow shock

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Earth and Planetary Physics 2024年第2期8卷 326-337页

作者： GuoQiang Wang SuDong Xiao MingYu Wu yuanqiang Chen TieLong Zhang Institute of Space Science and Applied Technology Harbin Institute of TechnologyShenzhen 518055China Shenzhen Key Laboratory of Numerical Prediction for Space Storm Harbin Institute of TechnologyShenzhen 518055China Mengcheng National Geophysical Observatory University of Science and Technology of ChinaHefei 230026China Chinese Academy of Sciences Center for Excellence in Comparative Planetology Hefei 230026China Space Research Institute Austrian Academy of SciencesGrazAustria

Magnetic holes are magnetic depression structures that exist widely in many plasma *** magnetic holes with durations of>1 s in the solar wind at Mercury’s orbit have drawn much attention,but the properties of the magnetic holes with shorter durations are still ***,we investigate the magnetic holes with durations of 0.1-100 s in the upstream region of Mercury’s bow shock based on observations by the MESSENGER(MErcury Surface,Space ENvironment,GEochemistry,and Ranging)*** can be divided into two groups according to the distribution of their duration:small-duration magnetic holes(SDMHs,0.6 s).The duration of each group approximately obeys a log-normal distribution with a median of~0.25 s and 3 s,*** 1.7%(32.6%)of the SDMHs(LDMHs)reduce the magnetic field strength by more than 50%.For both groups,some structures have a linear or quasi-linear polarization,whereas others have an elliptical *** magnetic hole events in both groups tend to have a higher rate of occurrence when the interplanetary magnetic field strength is *** occurrence rates are also affected by Mercury’s foreshock,which can increase(decrease)the occurrence rate of the SDMHs(LDMHs).This finding suggests that Mercury’s foreshock might be one source of the SDMHs and that the foreshock can destroy some *** observations suggest that a new group of magnetic holes with durations of<0.6 s exist in the upstream region of Mercury’s bow shock.

关键词： magnetic hole solar wind Mercury foreshock

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维普期刊数据库

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Lysosomes are Beneficial for(stearoyl) Gemcitabine-incorporated Solid Lipid Nanoparticles to Overcome Tumor Cell Chemoresistance

Lysosomes are Beneficial for(stearoyl) Gemcitabine-incorpora...

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第十三届全国免疫学学术大会

作者： Yanchun Shi Zhe Chen yuanqiang zheng zhengrong Cui Inner Mongolia Key Laboratory of Molecular Biology School of Basic Medical Sciences Inner Mongolia Medical University Division of Molecular Pharmaceutics and Drug Delivery College of Pharmacy The University of Texas at Austin

Despite recent advances in targeted therapies and immunotherapies, chemotherapy using cytotoxic agents remains an indispensable modality in cancer ***, there has been a growing emphasis in using nanomedicine in cancer chemotherapy, and several nanomedicines have already been used clinically to treat *** is evidence that for mulating small molecular cancer chemotherapeutic agents into nanomedicines significantly modifies their pharmacokinetics and often improves their ***, cancer cells often develop resistance to chemotherapy, and formulating anticancer drugs into nanomedicines also helps overcome *** this review, we briefly describe the different classes of cancer chemotherapeutic agents, their mechanisms of action and resistance, and evidence of overcoming the resistance using *** then emphasize on gemcitabine and our experience in discovering the unique(stearoyl) gemcitabine solid lipid nanoparticles that are effective against tumor cells resistant to gemcitabine and elucidate the underlying mecha *** seems that lysosomes, which are an obstacle in the delivery of many drugs, are actu ally beneficial for our(stearoyl) gemcitabine solid lipid nanoparticles to overcome tumor cell resistance to gemcitabine.

关键词： gemcitabine chemoresistance chemotherapeutic agents nanomedicine

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