1.Changing trends in the development of the thymus in duckling
Jing FANG ; Hengmin CUI ; Xi PENG ; Min HE
Chinese Journal of Veterinary Science 2008;28(5):544-548
By using routine H. E method and measuring the absolute weight and growth index of thymus and ANAE positive ratios, 1 to 49 days Tianfu duckling were selected to study the changing trends in the normal development of the thymus. The results are as follows:the thymus of duckling develops slowly from 1 to 14 days,and quickly from 21 to 35 days,then steadily from 35 to 49 days. By the 28th day,most T-lymphocytes have well differentiated and the thymus has reached its maturation. There are three types of thymie corpuscles whose functions have been discussed.
2.Bioinformatic prediction and functional characterization of human KIAA0100 gene
He CUI ; Xi LAN ; Shemin LU ; Fujun ZHANG ; Wanggang ZHANG
Journal of Pharmaceutical Analysis 2017;7(1):10-18
Our previous study demonstrated that human KIAA0100 gene is a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online software;Secondly, human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signal peptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.
3.Preparation of monoclonal antibody against human KIAA0100 protein and Northern blot analysis of human KIAA0100 gene
He Cui ; ?Xi Lan ; ?Shemin Lu ; ?Fujun Zhang ; ?Wanggang Zhang
Journal of Pharmaceutical Analysis 2017;7(3):190-195
Monoclonal antibodies (MAbs) are important tools for the study of proteins′ function and structure. But there has been no report on the preparation of MAbs against human KIAA0100 protein up to date. Here, first, we generated the mouse MAb against human KIAA0100 protein using purified recombinant 6×Histidinc (6×His)- tagged human KIAA0100 protein segment (1557–2234) as an antigen; then, the mRNA expression of human KIAA0100 gene was detected in U937 cells using Northern blot analysis. The results showed that the mouse MAb against human KIAA0100 protein could sensitively recognize the human KIAA0100 protein using Western blot analysis and immunocytochemistry analysis. Besides, Western blot analysis revealed that human KIAA0100 gene possibly encoded two different protein products (254 kDa and < 250 kDa) in U937 cells. Moreover, Northern blot analysis confirmed that human KIAA0100 gene might produced two different mRNA products (6000–10000 bp and 5000–6000 bp) in U937 cells. The results provide a basis for large-scale production of the MAb against human KIAA0100 protein, which will be useful for the study of human KIAA0100 protein′s function/structure and MAb-targeted drugs in the future.
4.Growth and histological development of bursa of Fabricius in ducklings
Jing FANG ; Hengmin CUI ; Min HE ; Xi PEN
Chinese Journal of Veterinary Science 2008;28(11):1301-1305,1309
By measuring the absolute weight and growth index of the bursa of Fabricius and using routine H.E method,the growth and histological development of the bursa in 1 to 49 days duckling were studied.The results showed that the absolute weight of the bursa increased as the ducklings grew.The growth index of the bursa reached its peak at 21 days.The height and width of both the large plica and projection formed by the follicle-associated epithelium(FAE)enlarged gradually,the same as the lymphoid follicle area and cortex width.The results indicated that the bursa of Fabricius in duckling develops slowly from 1 to 14 days,and quickly from 14 to 35 days(especially from 21 to 28 days),then steadily from 35 to 49 days.Histologically,the duckling bursa has approached its maturation by the 28th day.
5.Construction and biological identification of the transfectant cells expressing ??TCR with specific CDR3 sequence
Xueyan XI ; Wei CAO ; Lianxian CUI ; Wei HE
Chinese Journal of Immunology 2001;0(10):-
Objective:To construct of transfectant cells expressing ??TCR with specific CDR3 sequence.Methods:Specific CDR3 region sequence of DBS4.3,a known ??T cell clone,was inserted into ?9 and ?2 chain to substitute original CDR3 sequence using overlapping PCR method.After the full-length ?9 and ?2 chains were ligated with expression vector pREP7and pREP9 respectively,they were co-trasfected into a cell line of human Jurkat cells with TCR ? chain gene-defective mutant J.RT3-T3.5.When the transfectant cells expressing specific ??TCR were stimulated by antigen,production of IL-2 was detected by ELISA and Realtime PCR.Results:By ELISA and Realtime PCR,it was exhibited that the transfectant cells expressing DBS4.3 specific ??TCR secreted IL-2 under stimulation of iso-butylamine and anti ??TCR antibody.Conclusion:The transfectant Jurkat cells expressing ??TCR with specific CDR3 sequence are successfully constructed.It provides a platform for the research of recognition mechanism of ??TCR.
6.Characterization and Investigation of Reduction Capacity of Hydrophilic Organic Matter from Compost and its Influence Factors
Dongyu CUI ; Xiaosong HE ; Beidou XI ; Wenbing TAN ; Ying YUAN ; Rutai GAO
Chinese Journal of Analytical Chemistry 2015;(2):218-225
Reduction capacity ( RC ) is an important index to evaluate the redox ability of dissolved organic matter. In order to determine the RC, hydrophilic organic fractions ( HyI ) isolated from dissolved organic matter extracted from the uncomposted and composted samples were used as electron donators and mediators, and three kinds of irons were chosen as electron acceptors. The results showed that, the RC values from the composted sample were 15. 88, 13. 41 and 51. 45 mmol e -/mol C for the electron acceptors Fe2(SO4)3, Fe(NO3)3 and FeCit, respectively, which were higher than the corresponding values (13. 45, 11. 77 and 43. 16 mmol e-/mol C) from the uncomposted sample. The electron acceptor type shows a dramatic influence on the RC value of HyI. The RC value determined by FeCit was obviously higher than that measured using Fe2( SO4 ) 3 and Fe( NO3 ) 3 , and the microbial reducing capacity of the HyI was lower than the corresponding native reducing capacity. By analyzing the special absorbencies ( SUVA254 and SUVA280 ) , absorbance ratios ( A2/A3 and A4/A6 ) and integrated area from UV-vis spectra, it can be found that the RC was affected by aromatic degree, unsaturated conjugated structure, and molecular weight. Excitation-emission matrix spectra coupled with regional integration analysis showed that the relative content of humic-like substances ( humic-like acids and fulvic-like acids) was the main factor influencing the RC value of HyI. The results obtained can be used to characterize the redox properties of HyI, and reveal its role in the transformation and degradation of pollutants during composting.
7.Characteristic Study of Dissolved Organic Matter for Electron Transfer Capacity During Initial Landfill Stage
Chao YANG ; Xuaosong HE ; Beudou XI ; Cauhong HUANG ; Dongyu CUI ; Rutau GAO ; Wenbung TAN ; Huu ZHANG
Chinese Journal of Analytical Chemistry 2016;44(10):1568-1574
To unvestugate the evolutuon law and unfluenced factors of dussolved organuc matter for electron transfer capacuty durung unutual landfull stage, dussolved organuc matter ( DOM ) was extracted from landfull wastes at dufferent depth. Shewanella oneudensus MR-1 and cutrate uron ( FeCut ) were respectuvely used as electron donor and electron acceptor to measure electron donatung capacuty, electron acceptung capacuty and electron shuttlung capacuty. Afterwards, the unfluenced factors of electron transfer capacuty were studued by analyzung spectral unformatuon. The results showed that proteun-luke components and humuc-luke components were able to transfer electrons, and they also accepted electrons from mucroorganusms. Electron donatung capacuty and electron acceptung capacuty uncreased furstly and then decreased. However, the electron shuttlung capacuty uncreased persustently durung the landfull process. Proteun-luke components were the maun components of dussolved organuc matter durung the unutual landfull stage, and ut was maunly responsuble for the electron donorung capacuty and electron acceptung capacuty of DOM. Electron shuttlung capacuty resulted from humuc-luke components durung the cycluc redox process. Electron shuttlung capacuty persustently uncreased durung the landfull process based on humuc-luke components generated durung the stage.
8.Imaging findings of soft tissue infections in AIDS(report of 3 cases)
Cui-Yu JIA ; Xuan ZHAO ; Yong DUAN ; Ning HE ; Chun-Wang YUAN ; Xiao-Xi MAO ; Wei WANG ; Da-Wei ZHAO ;
Chinese Journal of Radiology 2001;0(03):-
Objective To evaluate X-ray,CT and MRI findings of soft tissue infections in AIDS. Methods Three cases of soft tissue infections with AIDS were retrospectively analyzed by comparing the imaging findings with pathological results.All patients were performed MRI,X-ray was in 1 case,CT was in 1 case.Results Cellulitis was in 1 case:MRI showed extended thickening of subcutaneous tissues, ill-defined hypointense areas on T_1WI and hyperintensity on T_2WI,and reticular pattern on GRE. Necrotizing fasciitis was in 1 case:MRI showed obvious thickening of subcutaneous tissues and deep fasciae, abnormally increased signal intensity on T_1 and T_2WI.Fluid collections were within muscles and muscles interval on fat-suppressed T2 WI.Tuberculosis was in 1 case:CT demonstrated multiple low density areas in the subcutaneous tissues and clear peripheral rim enhancement.MRI appeared hypointense on T_1WI and hyperintensity on T_2WI,and peripheral rim enhancement following gadolinium injection.Conclusion Infections of soft tissue are common complication in patients with AIDS,radiology is important in early diagnosis and treatment planning in this population.
9.Effect of small nuclear ribonucleoprotein polypeptide A on the malignant biological behavior of hepatocellular carcinoma cells and its mechanism
YAO Menglin ; WANG Ruhua ; CUI Xiaomeng ; CHEN Yifei ; GUO Dan ; HE Shuixiang ; LI Yarui
Chinese Journal of Cancer Biotherapy 2023;30(12):1074-1081
[摘 要] 目的:探究小核核糖核蛋白多肽A(SNRPA)在肝细胞癌(HCC)组织和细胞中的表达及其调控HCC细胞HepG2和Hep3B恶性生物学行为的作用及其机制。方法: 数据库分析SNRPA在泛癌组织中的表达及其与病理分期、HCC患者预后的相关性。常规培养HepG2和Hep3B细胞,将si-NC,si-SNRPA#1、si-SNRPA#2转染HepG2和Hep3B细胞,实验分为si-NC组、si-SNRPA#1组和si-SNRPA#2组;将SNRPA-vector和SNRPA-oe载体转染LO2细胞,分为SNRPA-vector组和SNRPA-oe组。qPCR法检测正常肝细胞和肝癌细胞以及转染各组HepG2和Hep3B细胞中SNRPA mRNA的表达,MTT法、Transwell法和WB法分别检测转染后各组HepG2和Hep3B细胞的增殖、迁移和侵袭能力以及EMT相关蛋白表达的变化。结果: 数据库分析显示,SNRPA mRNA在多数肿瘤组织中均呈高表达(均P<0.001)且与病理分期有关联(P<0.05或P<0.01)。SNRPA在HCC组织和细胞中均呈高表达(P<0.05或P<0.01),且与HCC患者的预后有关联(P<0.01)。敲减SNRPA表达明显抑制HepG2和Hep3B细胞增殖(P<0.05或P<0.01)而过表达SNRPA则能促进LO2细胞增殖(P<0.01),敲减SNRPA表达明显抑制HepG2和Hep3B细胞的迁移和侵袭能力(均P<0.01),明显促进E-cadherin的表达上调(P<0.01),而抑制N-cadherin、vimentin的表达(P<0.01)。结论: SNRPA在HCC组织及细胞中呈明显高表达,其可能通过调控上皮间质转化(EMT)进程进而促进HepG2和Hep3B细胞的增殖、迁移和侵袭。
10.A new method to obtain molecular marker of sequence-tagged site (STS) of Panax ginseng and P. quinquefolius.
Guang-hong CUI ; Lu-qi HUANG ; Xiao-jing TANG ; Xi-rong HE ; Xin LI
China Journal of Chinese Materia Medica 2007;32(11):1012-1015
OBJECTIVESearching a new molecular method to authenticate Panax ginseng and P. quenquefolium.
METHODSingle primers based on rDNA sequences of Panax species were designed to obtain polymorphic bands of P. ginseng and P. quinquefolius and then sequenced. Four PCR primers (two forword and two reverse primers) specific to P. ginseng and P. quinquefolius were designed.
RESULTPrimer Pg-6F, Pg-479R only amplified 474 bp band for P. ginseng and primer Pq-442F, Pq-658R only amplified 217 bp band for P. quinquefolius. It is indicated that the four primers could serve as specific STS primers for Panax species.
CONCLUSIONA new way to obtain STS primers of Panax species was established. This method is more quick and efficient than SCAR-PCR method and can serve as a model to obtain molecular markers for other Chinese material medica.
Base Sequence ; DNA Primers ; DNA, Plant ; genetics ; DNA, Ribosomal ; genetics ; Genetic Markers ; genetics ; Molecular Sequence Data ; Panax ; classification ; genetics ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; methods ; Sequence Analysis, DNA ; Sequence Tagged Sites ; Species Specificity