61 results on '"Xiao-Dong Xi"'
Search Results
2. [Promoting Effect of Rat
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Jian-Hua, Mao, Yan, Shen, Zheng, Ruan, and Xiao-Dong, Xi
- Abstract
To explore the effects of the rathHC745, hHC1690, humanIn 293T cells, the coagulation activity of FVIII was significantly enhanced when hHC745 and hHC1690 combined with rLC compared with them combined with hLC. The FVIII activity of hHC745+rLC was about 4.6 times higher than that of hHC745+hLC, while hHC1690+rLC was about 2.9 times higher than that of hHC1690+hLC. The data from ELISA showed that there was no significant difference in FVIII antigen expression when hHC745 and hHC1690 combined with rLC and hLC. The specific activity of hHC745+rLC increased to about 4.1 times compared with hHC745+hLC, while that of hHC1690+rLC increased to about 2.8 times compared with hHC1690+hLC. In HA mice administrated with hydrodynamic injection, the FVIII activity of hHC745+rLC and hHC1960+rLC was significantly higher than that of hHC745+hLC and hHC1690+hLC at comparable expression level. The FVIII activity of hHC745+rLC was significantly higher than that of hHC745+hLC, increasing to about 5.1 times, while, that of hHC1690+rLC increasing to about 2.1 times than hHC1690+hLC. ELISA results also showed that there was no significant difference in FVIII antigen expression when hHC745 and hHC1690 combined with rLC and hLC. The specific activity of hHC745+rLC increased to about 4.2 times compared with hHC745+hLC, while that of hHC1690+rLC increased to about 1.8 times compared with hHC1690+hLC. In addition, the activity of hHC1690 combined with rLC was significantly higher than that of hHC745 combined with rLC.rLC can significantly enhance the coagulation activity of FVIII when co-expressed with hHC of different length including hHC745 and hHC1690.腺相关病毒载体血清型8递送的大鼠研究大鼠将hHC745、hHC1690、人在293T细胞中,hHC745和hHC1690与rLC共表达时的FVIII活性显著高于与hLC共表达时的活性,分别提高到约4.6倍和2.9倍;ELISA结果表明hHC745和hHC1690与hLC和rLC共表达时的FVIII抗原表达无统计学差异;根据aPTT测得的FVIII活性与ELISA测得的FVIII抗原表达量计算的比活性结果显示,hHC745与rLC共表达时的比活性提高到与hLC共表达时的比活性的约4.1倍,而hHC1690与rLC共表达时的比活性提高到与hLC共表达时的约2.8倍。在高压注射的HA小鼠中,hHC745和hHC1690与rLC共表达时的FVIII活性显著高于与hLC共表达时的活性,活性分别提高到约5.1倍和2.1倍;ELISA结果同样显示,hHC745和hHC1690与hLC和rLC共表达时FVIII抗原表达无统计学差异;比活性结果表明,hHC745和hHC1690与rLC共表达时的比活性较与hLC共表达时的比活性显著提高,分别提高到约4.2倍和1.8倍;hHC1690与rLC共表达时的FVIII活性显著高于hHC745与rLC共表达时的活性。.rLC可以显著提高不同长度
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- 2022
3. The role of SLC26A8 homozygous variants in male infertility and flagellum abnormalities
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Bin Mao, Sha-Sha Zhang, Liang-Tao Zhao, Hui-Rong Huang, Feng-Die Ma, Wen-Jing Shi, Shan Li, Zi-Yan Nie, Han-Dan Xiao, Xiao-Dong Xie, and Pei-Qiang Li
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Diseases of the genitourinary system. Urology ,RC870-923 - Published
- 2024
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4. Inherent hepatocytic heterogeneity determines expression and retention of edited
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Qiang, Wang, Lin, Zhang, Guo-Wei, Zhang, Jian-Hua, Mao, Xiao-Dong, Xi, Lu, Jiang, Gang, Lv, Jing, Lu, Yan, Shen, Zhu, Chen, Jiang, Zhu, and Sai-Juan, Chen
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Factor IX ,Gene Editing ,Enhancer Elements, Genetic ,Mutation ,Hepatocytes ,Clustered Regularly Interspaced Short Palindromic Repeats ,Dependovirus ,Biological Sciences ,Promoter Regions, Genetic ,Hemophilia B ,Alleles ,Transcription Factors - Abstract
Infusing CRISPR/donor-loaded adeno-associated viral vectors (AAV/CRISPR) could enable in vivo hepatic gene editing to remedy hemophilia B (HB) with inherited deficiency of clotting factor IX (FIX). Yet, current regimens focus on correcting HB with simple mutations in the coding region of the F9, overlooking those carrying complicated mutations involving the regulatory region. Moreover, a possible adverse effect of treatment-related inflammation remains unaddressed. Here we report that a single DNA cutting-mediated long-range replacement restored the FIX-encoding function of a mutant F9 (mF9) carrying both regulatory and coding defects in a severe mouse HB model, wherein incorporation of a synthetic Alb enhancer/promoter-mimic (P2) ensured FIX elevation to clinically meaningful levels. Through single-cell RNA sequencing (scRNA-seq) of liver tissues, we revealed that a subclinical hepatic inflammation post-AAV/CRISPR administration regulated the vulnerability of the edited mF9-harboring host cells to cytotoxic T lymphocytes (CTLs) and the P2 activity in a hepatocytic subset–dependent manner via modulating specific sets of liver-enriched transcription factors (LETFs). Collectively, our study establishes an AAV/CRISPR-mediated gene-editing protocol applicable to complicated monogenetic disorders, underscoring the potentiality of improving therapeutic benefits through managing inflammation.
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- 2021
5. Identification of the pathogen of hepatopancreas necrosis syndrome (HPNS) in Litopenaeus vannamei and evaluation of the effect of plant extracts against HPNS
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Ying Zhou, Yu-Tong Ji, Zi Wei, Shuang Wang, Xiao-Dong Xie, Xin-Yu Chen, Ying-Yi Wei, Li-Ji Xie, Zhi-Xun Xie, Jia-Xun Feng, Yong-Zhen Zhao, Ting-Jun Hu, and Mei-Ling Yu
- Subjects
Hepatopancreas necrosis syndrome (HPNS) ,Litopenaeus vannamei ,Plant extracts ,Vibrio ,Proteus vulgaris ,Microbiology ,QR1-502 ,Veterinary medicine ,SF600-1100 - Abstract
Hepatopancreatic necrosis syndrome (HPNS) has recently become a major epidemic in shrimp farms, resulting in enormous economic loss. However, its etiology remains controversial. Diseased Litopenaeus vannamei (L. vannamei), collected from shrimp farms, were analyzed for pathological diagnosis, bacterial isolation, and identification. After that, the effects of the three plant extracts on the primary pathogens of HPNS were analyzed in vitro and in vivo. The results revealed that the primary pathogens of HPNS were Vibrio parahaemolyticus (V. parahaemolyticus) with ToxR and TLH gene, V. cholerae with ompW gene, and Proteus vulgaris (P. vulgaris) with rpoA gene. These three bacterial strains exhibited extensive antibiotic resistance. The “Chaiyinhugan” mixture, the “Sanhuanglian” mixture, and the Polygonum hydropiper flavonoid extract inhibited the growth of all three dominant strains. Additionally, the three plant extracts reduced HPNS shrimp mortality, improved shrimp immunity, and attenuated hepatopancreatic pathology. Among them, the Polygonum hydropiper flavonoid extract displayed better effects and could be applied to prevent and treat HPNS in producing and breeding L. vannamei. In conclusion, V. parahaemolyticus, V. cholerae, and P. vulgaris are the major pathogens causing HPNS. Herbal extracts such as Polygonum hydropiper flavonoid extract can be applied to prevent and treat HPNS.
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- 2024
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6. Lymphadenopathy in POEMS syndrome: a correlation between clinical features and imaging findings
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Xiao-Feng, Shi, Shu-Dong, Hu, Li-Li, Wu, Xiao-Yan, Chen, Jian-Nong, Wu, Xian-Qiu, Yu, Dong-Ya, Li, Min, Chen, Yi-Chen, Liu, Yan, Zhu, and Xiao-Dong, Xi
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Original Article - Abstract
Lymphadenopathy is an important characteristic of POEMS syndrome, and a Castleman disease (CD)-like pathologic change in the lymph nodes is one of the major diagnostic criteria. However, the characteristics of lymphadenopathy in POEMS still have not been completely elucidated. The lymph node biopsies are available only for a small proportion of patients. A simple and safe way is needed to rule CD in or out. This study aimed to analyse the features of lymphadenopathy and estimate the role of imaging methods, including computed tomography (CT) and positron emission tomography-CT (PET/CT), in the diagnosis of lymphadenopathy in patients with POEMS syndrome. We conducted a retrospective analysis of 23 patients with confirmed POEMS syndrome. All of the patients received chest and abdominal CT scan and/or superficial ultrasound examinations. Four patients underwent PET/CT examinations, and 6 patients received lymph node biopsies. Enlarged lymph nodes (short diameter ≥ 1 cm) were found in 48% (11/23) of patients, but only 1 patient had an enlarged lymph node with a diameter ≥ 2 cm. Lymph nodes with CD-like pathologic changes from 2 patients showed increased maximum standard uptake values (SUV(max)) of (18)F-deoxyglucose ((18)FDG) on PET/CT, while lymph nodes with reactive pathologic changes from 2 other patients showed a normal metabolic PET/CT profile. The extent of lymph node enlargement in patients with POEMS was less than that in patients with CD per se. We draw the conclusion that most of the enlarged lymph nodes had diameters ≤ 2 cm, which is less than that in cases of CD per se and PET/CT may be helpful in determining whether enlarged lymph nodes are characterized by CD-like changes or not.
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- 2020
7. [Effect of Amino Acid Motifs in Integrin β3 Cytoplasmic Tail on αⅡbβ3-Mediated Cell function in 293T cell models]
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Dong-Ya, Li, Jian-Hua, Mao, Wei, Zhang, Xin-Jie, Chen, Bing, Xiao, Zheng, Ruan, Yun, Wang, Guo-Xiong, Jiang, Xiao-Feng, Shi, and Xiao-Dong, Xi
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Cricetulus ,HEK293 Cells ,Cricetinae ,Amino Acid Motifs ,Cell Adhesion ,Integrin beta3 ,Animals ,Humans ,CHO Cells ,Platelet Glycoprotein GPIIb-IIIa Complex ,Signal Transduction - Abstract
To establish 293T cell lines stably expressing Calpain-cleavage related α3 cytoplasmic tail mutants, and to explore the effect of amino acid motifs in integrin β3 cytoplasmic tail on αⅡbβ3-mediated cell function.293T cell lines stably co-expressing human wild type integrin αⅡb and full length β3 or mutant β3, including β3-ΔNITY (β3 cytoplasmic tail NITY motif deleted), β3-Δ754 (β3 cytoplasmic tail TNITYRGT motif deleted) and β3-Δ759 (β3 cytoplasmic tail RGT motif deleted) were established. Spreading and adhesion of these stable cell lines on immobilized fibrinogen were tested.293T-αⅡbβ3ΔNITY, 293T-αⅡbβ3Δ754, 293T-αⅡbβ3Δ759 and 293T-αⅡbβ3 cell lines were successfully established. Compared with the 293T cells, 293T-αⅡbβ3 cells which expressed full β3, possessed well adhesion and spread ability on immobilized fibrinogen, suggesting it can be as a surrogate for platelet. Compared with 293T-αⅡbβ3 cells, the 293T-αⅡbβ3ΔNITY cells showed a partial impairment of adhesion and spreadability on immobilized fibrinogen. while the 293T-αⅡbβ3Δ754 cells and 293T-αⅡbβ3Δ759 cells failed to adhere or spread on immobilized fibrinogen.To the cell spreading function mediated by integrin β3, RGT motif is vital, while NITY can be dispensable. These established 293T cell lines stably expressing different β3 mutants provide a solid basis for a further analysis of mass spectrometry.
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- 2019
8. [Role and Mechanism of the Interaction of BCR-ABL with E3 Ligase c-CBL in Targeting Therapy of Chronic Myelogenous Leukemia]
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Jian-Hua, Mao, Qiu-Hua, Huang, Ping, Liu, Cheng, Luo, and Xiao-Dong, Xi
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive ,Ubiquitin-Protein Ligases ,Fusion Proteins, bcr-abl ,Humans ,Proto-Oncogene Proteins c-cbl ,Phosphorylation ,Arsenic ,HeLa Cells - Abstract
To explore the interaction domains between BCR-ABL and E3 liagase c-CBL, so as to reveal the structure-basis for the arsenic to treat chronic myelogenous leukemia(CML).The interactional interface of BCR-ABL and c-CBL was simulated and analyzed according to the available structure model. Based on the structural information, the WT and mutant Migr1-BCR-ABL-GFP (ΔSH2,ΔTyrKC,ΔSH2/TyrKC (S/H) and pFlag-c-CBL (ΔRF) were constructed and co-transfected into the 293T and HeLa cells. The co-immunoprecipitation (Co-IP) was performed by using M2 beads (anti-Flag), anti-GFP antibody and protein A beads, and the interaction was identified by using GFP and M2 antibody, respectively. Moreover, the colocalization of BCR-ABL and c-CBL was further evaluated by using immunofluorescent(IF) assay in transfected HeLa cells.Co-IP demonstrated that the TyrKC domain of BCR-ABL was primarily involved in the interaction with c-CBL, while both the SH2 domain of BCR-ABL and the RF domain of c-CBL also participated in the interaction to a certain degree, which were consistent with the structure-based simulation. IF elucidated that the colocalization of BCR-ABL and c-CBL was almost entirely vanished when the deleted TyrKC domain of BCR-ABL was co-transfected with c-CBL, which were elegantly coincident with the results from Co-IP.The TyrKC domain of BCR-ABL is sufficient and necessary to mediate the interaction between BCR-ABL and c-CBL, the SH2 domain of BCR-ABL and the RF domain of c-CBL are also involved in the association between the two proteins. It suggests that the association of BCR-ABL and c-CBL can modulate the stability and degradation of BCR-ABL, thus illustrating the molecular mechanisms of the targeting therapy for CML by arsenic.
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- 2017
9. An atom-efficient and powerful method for direct esterification of silyl ethers catalyzed by HClO4–SiO2
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Yi-Nan Shu, Xi Chen, Qing-Shan Zhang, Xiao-Dong Xi, Yunzheng Li, Qin-Pei Wu, Ti-Jian Du, and Hai-Xia Liu
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chemistry.chemical_classification ,Silylation ,Silica gel ,Aryl ,Organic Chemistry ,Biochemistry ,Catalysis ,Silyl ether ,Acetic anhydride ,chemistry.chemical_compound ,chemistry ,Drug Discovery ,Organic chemistry ,Perchloric acid ,Alkyl - Abstract
An efficient and convenient procedure for direct esterification of alkyl and aryl silyl ethers with Ac2O and a catalyst system of perchloric acid immobilized on a silica gel (HClO4–SiO2) has been developed. The silyl protecting groups are directly replaced by acetyls and the protecting groups themselves are transformed into acetates as the sole byproducts, which can be readily recovered and converted back to silylchlorides, the original protecting agents, thus minimizing wastes.
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- 2011
10. Synthesis and antitumor activity of novel 2′,3′-diethanethio-2′,3′,5′-trideoxy-5′-triazolonucleoside analogues
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Qin-Pei Wu, Hongquan Yin, Yunzheng Li, Yan-Hong Liu, Xiao-Dong Xi, Ning-Ning Liu, Qing-Shan Zhang, and Jin-Lan Yu
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Pharmacology ,Antitumor activity ,Stereochemistry ,Organic Chemistry ,Triazole ,Cancer therapy ,Antineoplastic Agents ,Nucleosides ,Tumor cells ,General Medicine ,Inhibitory Concentration 50 ,chemistry.chemical_compound ,chemistry ,Cell culture ,Cell Line, Tumor ,Drug Discovery ,Humans ,Inhibitory concentration 50 ,Nucleoside - Abstract
A series of novel 2',3'-diethanethio-2',3',5'-trideoxy-5'-triazoloribonucleosides was synthesized in excellent yields and their antitumor activity was evaluated. These nucleoside analogues with aromatic substituted triazole rings showed significantly improved activity towards a broad range of tumor cell lines and those without arene substitutes were inactive.
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- 2010
11. A Vicinal Acyloxy Group Participation SN2 Reaction of Thiol Nucleophiles in the Formation of Thioacetals
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Qing-Shan Zhang, Qinpei Wu, Xi Chen, Hui Li, and Xiao-Dong Xi
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chemistry.chemical_classification ,chemistry.chemical_compound ,Nucleophile ,Chemistry ,Ethanethiol ,Thioacetal ,Acetal ,Thiol ,SN2 reaction ,Organic chemistry ,General Chemistry ,Protecting group ,Acyl group - Abstract
Thioacetalization of acyl protected furanosides led to products with an ethanethiol group at C-2 and 3-O-acetyl- 1,2-di-O-isopropylidene-D-furanoses were converted into corresponding thioacetals with two ethanethiol groups at both C-2 and C-3 positions under the standard thioacetalization conditions. All products were characterized by 1H NMR, 13C NMR and HRMS data. X-ray structure analysis indicates that the vicinal acyloxy group is stereoselectively substituted by ethanethiols. The supposed mechanisms for these two kinds of transformations were presented.
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- 2009
12. RGT, a synthetic peptide corresponding to the integrin β3 cytoplasmic C-terminal sequence, selectively inhibits outside-in signaling in human platelets by disrupting the interaction of integrin αIIbβ3 with Src kinase
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Hongchen Liu, Xiaoyu Su, Xiao-dong Xi, Jingsong Yan, Xiaoping Du, Jian-Qing Mi, Panagiotis Flevaris, Xuefeng Wang, Yuanjing Lu, Sai-Juan Chen, Zheng Ruan, and Nelly Kieffer
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Immunology ,Integrin ,Clot Retraction ,Platelet Glycoprotein GPIIb-IIIa Complex ,Biochemistry ,CD49c ,Collagen receptor ,Platelet Adhesiveness ,Humans ,Phosphorylation ,Dose-Response Relationship, Drug ,biology ,Integrin beta3 ,Fibrinogen binding ,Cell Biology ,Hematology ,Molecular biology ,Peptide Fragments ,Cell biology ,src-Family Kinases ,Integrin alpha M ,biology.protein ,Integrin, beta 6 ,Protein Binding ,Signal Transduction ,Proto-oncogene tyrosine-protein kinase Src - Abstract
Mutational analysis has established that the cytoplasmic tail of the integrin β3 subunit binds c-Src (termed as Src in this study) and is critical for bidirectional integrin signaling. Here we show in washed human platelets that a cell-permeable, myristoylated RGT peptide (myr-RGT) corresponding to the integrin β3 C-terminal sequence dose-dependently inhibited stable platelet adhesion and spreading on immobilized fibrinogen, and fibrin clot retraction as well. Myr-RGT also inhibited the aggregation-dependent platelet secretion and secretion-dependent second wave of platelet aggregation induced by adenosine diphosphate, ristocetin, or thrombin. Thus, myr-RGT inhibited integrin outside-in signaling. In contrast, myr-RGT had no inhibitory effect on adenosine diphosphate-induced soluble fibrinogen binding to platelets that is dependent on integrin inside-out signaling. Furthermore, the RGT peptide induced dissociation of Src from integrin β3 and dose-dependently inhibited the purified recombinant β3 cytoplasmic domain binding to Src-SH3. In addition, phosphorylation of the β3 cytoplasmic tyrosines, Y747 and Y759, was inhibited by myr-RGT. These data indicate an important role for β3-Src interaction in outside-in signaling. Thus, in intact human platelets, disruption of the association of Src with β3 and selective blockade of integrin αIIbβ3 outside-in signaling by myr-RGT suggest a potential new antithrombotic strategy.
- Published
- 2008
13. A facile one-pot procedure for the transformation of acetonides into diacetates catalyzed with Bi(OTf)3·xH2O
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Hai-Xia Liu, Ming-Xin Zhou, Yunzheng Li, Qin-Pei Wu, Qing-Shan Zhang, Di Song, Xiao-Dong Xi, and Yuan Wang
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Anomer ,Primary (chemistry) ,Chemistry ,Stereochemistry ,Organic Chemistry ,Acetal ,Homogeneous catalysis ,Biochemistry ,Chemical synthesis ,Catalysis ,chemistry.chemical_compound ,Transformation (genetics) ,Drug Discovery ,Organic chemistry - Abstract
The transformation of acetonides into the corresponding diacetates is often required in the synthetic chemistry. An efficient procedure for direct conversion of acetonides into diacetates in the presence of Bi(OTf) 3 · x H 2 O under mild conditions has been developed. Primary hydroxyl-acetonides could be selectively transformed into diacetates in the presence of anomeric acetonides and the anomeric acetonides could be tunably converted into 2-acetoxyisopropyl or diacetate groups.
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- 2008
14. Fibrinogen interaction of CHO cells expressing chimeric αIIb/αvβ3 integrin
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Juan-juan Chen, Liping Lin, Xiao-yu Su, Jian Ding, He Lu, and Xiao-dong Xi
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Pharmacology ,biology ,Chinese hamster ovary cell ,Protein subunit ,Integrin ,Fibrinogen binding ,General Medicine ,Molecular biology ,Integrin alpha M ,biology.protein ,Pharmacology (medical) ,Signal transduction ,Cell adhesion ,Glycoprotein IIb/IIIa - Abstract
Aim: The molecular mechanisms of the affinity regulation of αvβ3 integrin are important in tumor development, wound repairing, and angiogenesis. It has been established that the cytoplasmic domains of αvβ3 integrin play an important role in integrin-ligand affinity regulation. However, the relationship of structure-function within these domains remains unclear. Methods: The extracellular and transmembrane domain of αIIb was fused to the αv integrin cytoplasmic domain, and the chimeric α subunit was coexpressed in Chinese hamster ovary (CHO) cells with the wild-type β3 subunit or with 3 mutant β3 sequences bearing truncations at the positions of T741, Y747, and F754, respectively. The CHO cells expressing these recombinant integrins were tested for soluble fibrinogen binding and the cell adhesion and spreading on immobilized fibrinogen. Results: All 4 types of integrins bound soluble fibrinogen in the absence of agonist stimulation, and only the cells expressing the chimeric α subunit with the wild-type β3 subunit, but not those with truncated β3, could adhere to and spread on immobilized fibrinogen. Conclusion: The substitution αIIb at the cytoplasmic domain with the αv cytoplasmic sequence rendered the extracellular αIIbβ3 a constitutively activated conformation for ligands without the need of “inside-out” signals. Our results also indicated that the COOH-terminal sequence of β3 might play a key role in integrin αIIb/αvβ3-mediated cell adhesion and spreading on immobilized fibrinogen. The cells expressing αIIb/αvβ3 have enormous potential for facilitating drug screening for antagonists either to αvβ3 intracellular interactions or to αIIbβ3 receptor functions.
- Published
- 2008
15. [Effect of the Integrin β3 Cytoplasmic NITY Motif on α II bβ3-Mediated Cell Functions in CHO Cell Model]
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Ji-Chun, Yang, Xiao-Feng, Shi, Jian-Song, Huang, Zhang-Biao, Long, Bing, Xiao, Zheng, Ruan, and Xiao-Dong, Xi
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Cricetulus ,Cricetinae ,Integrin alpha2 ,Integrin beta3 ,Animals ,Fibrinogen ,Humans ,CHO Cells ,Platelet Glycoprotein GPIIb-IIIa Complex ,Protein Structure, Tertiary ,Signal Transduction - Abstract
OBJLECTIVE: To investigate the effect of integrin β3 cytoplasmic NITY motif on αIIbβ3-mediated cell functions.Stable Chinese hamster ovary (CHO) cell lines that co-express human wild type integrin αIIb and wild type β3 or mutant β3ΔNITY (β3 deleting cytoplasmic NITY motif) were established. Expression of αIIb and β3 were tested by Western blot and flow cytometry in CHO cell lines. Spreading and adhesion of stable cell lines on immobilized fibrinogen were examined. The co-immunoprecipitation was used to detect protein interactions.CHO-αIIbβ3, CHO-αIIbβ3ΔNITY cells were successfully established. The CHO cells transfected with wild type αIIbβ3 had the ability of adhesion and spreading. Compared with CHO-αIIbβ3 cells, CHO-αIIbβ3ΔNITY cells showed an impaired capacity of adhesion but no significant difference was observed in spreading of adhered cells. The co-immunoprecipitation showed that kindlin-2 associated with wild type integrin αIIbβ3. The β3ΔNITY mutation substantially reduced kindlin-2 association.Deletion of NITY motif causes an impaired ability of adhesion. The deletion mutation can suppress kindlin-2 binding to integrin β3, thereby partially inhibit the integrin β3 signaling.
- Published
- 2015
16. [Myr-RKEFAK Peptide Selectively Regulates Outside-in Signaling Transduction-related Functions in Human Platelets]
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Zhang-Biao, Long, Jian-Song, Huang, Xiao-Feng, Shi, Ji-Chun, Yang, Zheng, Ruan, Bing, Xiao, and Xiao-Dong, Xi
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Blood Platelets ,Platelet Adhesiveness ,Platelet Aggregation ,Integrin beta3 ,Fibrinogen ,Humans ,Peptides ,Signal Transduction - Abstract
To study the effect of interaction of the talin rod domain integrin binding site 2 with integrin β3 on platelet signal transduction.A peptide that mimics the membrane proximal α helix 6 residues R724 KEFAK729 of the integrin β3 cytoplasmic tails was designed and synthesized, to which the myristoylation was covalently linked to the N-terminal of the peptide enabling membrane penetration. The effects of myr-RKEFAK peptide on the typical platelet outside-in signaling ovent (stable adhesion and spreading on immobilized fibrinogen, aggregation, fibrin clot retraction) and inside-out signaling events (soluble fibrinogen binding) were tested.myr-RKEFAK peptide dose-dependently inhibited platelet stable adhesion and spreading on immobilized fibrinogen, irreversible aggregation, as well as fibrin clot retraction, but not soluble fibrinogen binding and reversible phase of platelet aggregation.The cell-penetrating peptide myr-RKEFAK causes an inhibitory effect on integrin β3 outside-in signaling-regulated platelets functions, but did not affect inside-out signaling-regulated platelets functions.
- Published
- 2015
17. Multimodal imaging and clinical characteristics of bone lesions in POEMS syndrome
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Xiao-Feng, Shi, Shu-Dong, Hu, Jun-Min, Li, Xian-Fu, Luo, Zhang-Biao, Long, Yan, Zhu, and Xiao-Dong, Xi
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Original Article - Abstract
POEMS syndrome is a rare plasmacyte-associated disease, one of the major diagnostic criteria of which is sclerotic bone lesion. To detect bone lesions in POEMS syndrome, which imaging method should be routinely applied and what characteristics they display are still unconfirmed. We analyzed clinical data and imaging characteristics of bone lesions in 22 patients with POEMS using multimodal methods, including conventional X-ray, computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography/computed tomography (PET/CT). Images on X-ray and CT exhibited plaque-like high-density for osteosclerotic lesions and punched-out low-density appearance for osteolytic ones. X-ray had advantage in detecting bone lesions in skull, extremity long bones, clavicle, and scapula, while CT could display sharp outline of lesions and was more sensitive than X-ray in detecting the small lesions. Osteosclerotic lesions on MRI demonstrated decreased signal intensity on both T1 and T2-weighted sequences, while osteolytic lesions or osteolytic part of mixed lesions showed high signal intensity on T2-weighted sequences. MRI had same sensitivity as CT, but with superiority in distinguishing the active osteolytic lesions from the osteosclerotic ones. PET-CT showed (18)F-FDG uptake was normal in the majority of osteosclerotic lesions, and slightly increased in mixed ones, but obviously elevated in osteolytic ones. PET/CT was less sensitive in detecting osteosclerotic lesions than in detecting osteolytic ones. In conclusion, to detect bone lesions in POEMS, conventional X-ray scan should be first performed, further followed by more sensitive CT or MRI. PET-CT is optional when the osteolytic lesions are suspected.
- Published
- 2015
18. Homoharringtonine deregulates MYC transcriptional expression by directly binding NF-κB repressing factor.
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Wei-Na Zhang, Bing Chen, Wen-Da Xi, Ying Lu, Jin-Yan Huang, Yue-Ying Wang, Jun Long, Song-Fang Wu, Yun-Xiang Zhang, Shu Wang, Si-Xing Li, Tong Yin, Min Lu, Xiao-Dong Xi, Jun-Min Li, Kan-Kan Wang, Zhu Chen, Sai-Juan Chen, and Xin-Jie Chen
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LEUKEMIA ,PROTEIN synthesis ,ENZYME inhibitors ,MYC proteins ,BINDING sites - Abstract
Homoharringtonine (HHT), a known protein synthesis inhibitor, has an anti-myeloid leukemia effect and potentiates the therapeutic efficacy of anthracycline/cytarabine induction regimens for acute myelogenous leukemia (AML) with favorable and intermediate prognoses, especially in the t(8;21) subtype. Here we provide evidence showing that HHT inhibits the activity of leukemiainitiating cells (Lin
- /Sca-1- /c-kit+ ; LICs) in a t(8;21) murine leukemia model and exerts a down-regulating effect on MYC pathway genes in human t(8;21) leukemia cells (Kasumi-1). We discovered that NF-κB repressing factor (NKRF) is bound directly by HHT via the second double-strand RNA-binding motif (DSRM2) domain, which is the nuclear localization signal of NKRF. A series of deletion and mutagenesis experiments mapped HHT direct binding sites to K479 and C480 amino acids in the DSRM2 domain. HHT treatment shifts NKRF from the nucleus (including nucleoli) to the cytoplasm by occupying the DSRM2 domain, strengthens the p65-NKRF interaction, and interferes with p65-p50 complex formation, thereby attenuating the transactivation activity of p65 on the MYC gene. Moreover, HHT significantly decreases the expression of KIT, a frequently mutated and/or highly expressed gene in t(8;21) AML, in concert with MYC down-regulation. Our work thus identifies a mechanism of action of HHT that is different from, but acts in concert with, the known mode of action of this compound. These results justify further clinical testing of HHT in AML. [ABSTRACT FROM AUTHOR]- Published
- 2019
- Full Text
- View/download PDF
19. Effects of different water quality regulators on growth performance, immunologic function, and domestic water quality of GIFT tilapia
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Liang-Gang Wang, Meng-Qian Liu, Xiao-Dong Xie, Yu-Bo Sun, Ming-Lin Zhang, Yi Zhao, Qi Chen, Yi-Qu Ding, Mei-Ling Yu, Zheng-Min Liang, Ting-Jun Hu, Wan-Wen Liang, and Ying-Yi Wei
- Subjects
Medicine ,Science - Abstract
Water quality regulation is widely recognized as a highly effective strategy for disease prevention in the field of aquaculture, and it holds significant potential for the development of sustainable aquaculture. Herein, four water quality regulators, including potassium monopersulfate (KMPS), tetrakis hydroxymethyl phosphonium sulfate (THPS), bacillus subtilis (BS), and chitosan (CS), were added to the culture water of Oreochromis niloticus (GIFT tilapia) every seven days. Subsequently, the effects of these four water quality regulators on GIFT tilapia were comprehensively evaluated by measuring the water quality index of daily growth-related performance and immune indexes of GIFT tilapia. The findings indicated that implementing the four water quality regulators resulted in a decrease in the content of ammonia nitrogen, active phosphate, nitrite, total organic carbon (TOC), and chemical oxygen demand (COD) in the water. Additionally, these regulators were found to maintain dissolved oxygen (DO) levels and pH of the water effectively. Furthermore, using these regulators demonstrated positive effects on various physiological parameters of GIFT tilapia, including improvements in final body weight, weight gain rate (WGR), specific growth rate (SGR), condition factor (CF), feed conversion ratio (FCR), spleen index (SI), hepato-somatic index (HSI), immune cell count, the activity of antioxidant-related enzymes (Nitric oxide, NO and Superoxide dismutase, SOD), and mRNA expression levels of immunity-related factors (Tumor Necrosis Factor-alpha, TNF-α and Interleukin-1 beta, IL-1β) in the liver and spleen. Notably, the most significant improvements were observed in the groups treated with the BS and CS water quality regulators. Moreover, BS and CS groups exhibited significantly higher serum levels of albumin (ALB) and total protein (TP) (P < 0.05), whereas the other indicators showed no significant difference (P > 0.05) compared to the control group. However, the KMPS and THPS groups of GIFT tilapia exhibited significantly higher serum levels of aspartate aminotransferase (AST), alanine transaminase (ALT), creatinine (CRE) and blood urea nitrogen (BUN) (P < 0.05), whereas they exhibited significantly decreased HSI (P < 0.05). In addition, the partially pathological observations revealed the presence of cell vacuolation, nuclear shrinkage, and pyknosis within the liver. In conclusion, these four water quality regulators, mainly BS and CS, could improve the growth performance and immunity of GIFT tilapia to varying degrees by regulating the water quality and then further increasing the expression levels of immune-related factors or the activity of antioxidant-related enzymes of GIFT tilapia. On the contrary, the prolonged use of KMPS and THPS may gradually diminish their growth-enhancing properties and potentially hinder the growth of GIFT tilapia.
- Published
- 2023
20. [Research progress on the effect of shear stress on thrombosis]
- Author
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Xiao-feng, Shi, Jian-song, Huang, Yu-lan, Zhou, Xiong-ying, Cui, Ping, Liu, and Xiao-dong, Xi
- Subjects
Humans ,Thrombosis ,Stress, Mechanical - Published
- 2014
21. [The binding mechanisms of F VIII Trp1707Ser mutation-associated inhibitor]
- Author
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Xi, Wu, Ye-ling, Lu, Qiu-lan, Ding, Jing, Dai, Xiao-dong, Xi, Hong-li, Wang, and Xue-feng, Wang
- Subjects
Male ,Young Adult ,Binding Sites ,Factor VIII ,Mutation ,Humans ,Exons ,Hemophilia A - Abstract
To investigate the binding mechanisms of FVIII Trp1707Ser mutation-associated inhibitor.The APPT, PT, TT, Fg and FVIII:C were detected to make phenotypic diagnosis of haemophilia A. Inhibitors titer were measured by Bethesda method. Long distance-PCR (LD-PCR) and sequence-specific PCR were adopted for screening the intron 22 and intron 1 inversions respectively. FVIII coding and boundary sequences were analyzed by direct DNA sequencing. Inhibitor was reacted with different segments of FVIII, including heavy chain and its components A1 and A2, light chain and its components A3, C1 and C2. Corrected test was used to measure the remaining F VIII:C (% ) by adding pooled normal plasmas. After labeling purified inhibitors with biotin, western blot was performed to further confirm the binding reactions between inhibitors and segments.The haemophilia A patient had mild deficiency of FVIII:C (1.1%) and had high FVIII inhibitor titer of 18.4 BU. A mutation c.97223CG in exon 14 of F8 gene resulted to p.Trp1707Ser was identified by DNA sequencing. Corrected test showed that the remaining F VIII:C was increased when inhibitors reacted with heavy chain and light chain, especially with heavy chain. The remaining FVIII:C was also increased in the A2 and C2 domain reactions. No significant differences were seen in the A1, A3 and C1 domain reactions. Antigen-antibody reaction bands were confirmed by western blots when degenerated B-domain deleted recombinant FVIII, A2 and C2 were used as antigens.The binding sites of FVIIITrp1707Ser mutation inhibitor were the A2 domain of heavy chain and C2 domain of light chain. The binding reaction with heavy chain was more intense.
- Published
- 2013
22. [Transgenic mouse models of the truncated platelet integrin β3 cytoplasmic tail established by stem cell transplantation]
- Author
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Xiong-Ying, Cui, Xiao-Feng, Shi, Jian-Song, Huang, Ping, Liu, Lan-Lan, Tao, Yu-Lan, Zhou, Zheng, Ruan, and Xiao-Dong, Xi
- Subjects
Blood Platelets ,Mice, Inbred C57BL ,Mice ,Retroviridae ,Genetic Vectors ,Hematopoietic Stem Cell Transplantation ,Integrin beta3 ,Animals ,Mice, Transgenic - Abstract
This study was purpose to establish the transgenic mouse models of the truncated platelet integrin β3 by retrovirus-infected hematopoietic stem cells (HSCs) transplantation and to provide the basis for further study of the role of integrin β3 cytoplasmic domain in platelet bi-directional signaling pathways. Wild-type β3, β3-Δ759 (R(760) GT(762) truncated β3) and β3-Δ754 (T(755) NITYRGT(762) truncated β3) cDNAs were subcloned into MSCV MigR1 retroviral vector bearing a GFP gene and packaged into infective retrovirus with BOSC23 cell strain. The bone marrow HSCs of the β3 deficient mice were infected by the retroviruses, and transplanted into lethally-irradiated wild type C57BL/6 mice. GFP positive rate and surface β3 expression of the recipients' platelets at 6 to 8 weeks after transplantation were detected by flow cytometry to evaluate the transgenic efficiency. The results showed that four kinds of transgenic mouse models including vector, wild-type β3, β3-Δ759 and β3-Δ754 were established successfully. GFP positive rates of transgenic mouse platelets ranged from 18% to 66% and the β3 expression of transgenic mouse reached heterozygote (β3(+/-) level of mouse). It is concluded that establishment of transgenic mouse models mediated by retrovirus-infected HSCs transplantation is a feasible, fast, and high throughput transgenic approach and laid a solid foundation for further research on the role of integrin β3 cytoplasmic domain for bi-directional signaling of platelets in vivo, and for the gene therapy of platelet disorders.
- Published
- 2013
23. [Functional study of abnormal fibrinogen caused by Arg275His mutation in fibrinogen γ chain]
- Author
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Jing-yi, Zhou, Xue-feng, Wang, Qiu-lan, Ding, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Male ,Phenotype ,Genotype ,Fibrinogens, Abnormal ,Mutation ,Fibrinogen ,Humans ,Female ,Afibrinogenemia ,Child ,Pedigree - Abstract
To investigate the function of abnormal fibrinogen in two inherited dysfibrinogenemia pedigrees.Routine coagulation tests were conducted in the probands and related family members. The antigen and activity levels of fibrinogen were detected by immunoturbidimetry assay and clauss assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes and antithrombin gene(AT3)were analyzed by PCR amplification and direct sequencing. Routine thrombelastography (TEG) test and functional fibrinogen TEG test were both used to make a comprehensive evaluation of coagulation status and functional fibrinogen level in patients. The molecular weights of the three peptides from fibrinogen were measured by Western blot. The function of abnormal fibrinogen was assessed by fibrinogen dynamic polymerization and fibrinolysis velocity.The coagulation routine tests were normal in two probands except for prolonged thrombin time (TT) and reptilase time (RT), as well as reduced activity levels of 0.5 g/L and 0.6 g/L fibrinogen, respectively. The antigen levels of fibrinogen were 2.32 g/L and 2.66 g/L in two probands, which were in the normal reference range. The genotype analysis showed that Arg275His in fibrinogen γ chain (γ Arg275His) existed in both probands and patients in these two pedigrees. Meanwhile, proband B's grandfather and aunt also carried heterozygote g.5876TC (Ser116Pro) mutation in AT3. The results of routine TEG test demonstrated that the α values of proband B and his father were close to and lower than the lower limit of reference range, respectively, while the MA values were normal in both of them. However, functional fibrinogen TEG test revealed obviously reduced MA value. All the probands and patients demonstrated prolonged lag-off time and reduced peak value in fibrinogen dynamic polymerization tests. Meanwhile, most of fibrin formed from the patients' plasma could not be dissolved completely by plasminogen (PLG) and urokinase-typeplasminogenactivator (u-PA) at a certain time.We first reported cases of inherited dysgibrinogenemia associated with inherited AT deficiency. γArg275His mutation caused the abnormal fibrinogen in terms of fibrin mono polymerization and possibly in fibrinolysis. Combined use of routine TEG test and functional fibrinogen TEG test with comprehensive analyses of the parameters in both tests could better evaluate the level of functional fibrinogen and predict the risk of hemorrhage and thrombosis in patients with inherited dysfibrinogenemia.目的 对两个遗传性异常纤维蛋白原血症家系的突变纤维蛋白原(Fg)进行功能研究。方法 常规筛查凝血功能;Fg抗原和活性分别用免疫比浊法和Clauss法测定;抽提DNA,对Fg 3个基因(FGA、 FGB和FGG)以及抗凝血酶基因(AT3)所有外显子及侧翼序列进行PCR扩增、测序及分析;采用常规血栓弹力图(TEG)和功能性Fg TEG检查对家系B先证者及其父亲进行凝血功能的综合评价及血浆功能性Fg评估;应用Western blot 检测血浆Fg肽链分子量;采用Fg动态聚集曲线和纤维蛋白溶解曲线实验检测血浆Fg的功能。结果 2例先证者凝血酶原时间(TT)和爬虫酶凝固时间(RT)明显延长,Fg活性仅为0.5 g/L和0.6 g/L,但其抗原均正常,分别为2.32 g/L和2.66 g/L。两个家系先证者均存在γ链Arg275His杂合突变,家系B先证者的祖父和姑母同时检出AT3 g.5876TC (Ser116Pro)杂合突变。家系B先证者及其父亲TEG检测结果中α值分别接近和低于正常参考值范围下限,但最大波幅(MA值)均为正常;在功能性Fg TEG检测中,MA值明显偏低。Fg动态聚集曲线中先证者和家系患者的起跳时间明显延长、峰值明显降低。纤维蛋白溶解曲线中多数患者的纤维蛋白在特定时间内不能被纤溶酶原完全溶解。结论 首次发现遗传性异常纤维蛋白原血症合并AT缺陷的患者。γ链 Arg275His突变使Fg在纤维蛋白单体聚合以及纤维蛋白溶解方面出现异常。联合应用常规TEG和功能性TEG检测,可以更好地评估异常纤维蛋白原血症患者Fg的功能。
- Published
- 2013
24. RNA-Seq and 16S rRNA Reveals That Tian–Dong–Tang–Gan Powder Alleviates Environmental Stress-Induced Decline in Immune and Antioxidant Function and Gut Microbiota Dysbiosis in Litopenaeus vannami
- Author
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Xiao-Dong Xie, Ying Zhou, Yu-Bo Sun, Shou-Li Yi, Yi Zhao, Qi Chen, Ying-Hong Xie, Mi-Xia Cao, Mei-Ling Yu, Ying-Yi Wei, Ling Zhang, and Ting-Jun Hu
- Subjects
Asparagus cochinchinensis (Lour.) Merr. ,Panax notoginseng (Burkill) F.H. Chen ex C.H. ,Litopenaeus vannamei ,antioxidant function ,gut microbiota ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Ammonia stress and nitrite stress can induce immune depression and oxidative stress in Litopenaeus vannami (L. vannamei). Earlier reports showed that L. vannamei immunity, resistance to ammonia stress, and resistance to nitrite stress improved after Tian–Dong–Tang–Gan Powder (TDTGP) treatment, but the mechanism is not clear. In this study, three thousand L. vannamei were fed different doses of TDTGP for 35 days and then subjected to ammonia and nitrite stress treatments for 72 h. Transcriptome and 16-Seq ribosomal RNA gene sequencing (16S rRNA-seq) were used to analyze hepatopancreas gene expression and changes in gut microbiota abundance in each group. The results showed that after TDTGP treatment, hepatopancreas mRNA expression levels of immunity- and antioxidant-related genes were increased, the abundance of Vibrionaceae in the gut microbiota was decreased, and the abundance of Rhodobacteraceae and Flavobacteriaceae was increased. In addition, after TDTGP treatment, the effects of ammonia and nitrite stress on the mRNA expression of Pu, cat-4, PPAF2, HO, Hsp90b1, etc. were reduced and the disruption of the gut microbiota was alleviated. In short, TDTGP can regulate the immunity and antioxidant of L. vannamei by increasing the expression levels of immunity- and antioxidant-related genes and regulating the abundance of Rhodobacteraceae and Flavobacteriaceae in the gut microbiota.
- Published
- 2023
- Full Text
- View/download PDF
25. [Phenotype and genotype analysis of two Chinese pedigrees with type 3 von Willebrand diseases]
- Author
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Lin-lin, Jiang, Xue-feng, Wang, Qiu-lan, Ding, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Hong-li, Wang, and Xiao-dong, Xi
- Subjects
Male ,Phenotype ,Adolescent ,Genotype ,Mutation ,von Willebrand Factor ,Humans ,Female ,von Willebrand Disease, Type 3 ,Pedigree - Abstract
To analyze the phenotype and genotype of two Chinese pedigrees with von Willebrand diseases, and to investigate the molecular pathogenesis.Bleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor-ristocetin cofactor (vWF:Rco), von Willebrand factor antigen (vWF:Ag), von Willebrand factor activity (vWF:A), von Willebrand factor collagen binding assay (vWF:CB) and multimer analysis were used for phenotype diagnosis. DNA was extracted. All of the 52 exons and exon-intron bounda ries of the VWF gene were amplified with polymerase chain reaction(PCR) and analyzed by direct sequencing.APTT and BT were prolonged. Plasma RIPA, vWF:Rco, vWF:Ag, vWF:A and vWF:CB was significantly decreased. No VWF multimer can be found by plasma VWF multimer analysis. Homozygous insertional mutation g.82888_82889insCATG in exon 17 was found in proband A. Compound heterozygous mutations g.94865 G to A (Trp856stop) in exon 20 and g.110698_110699delinsG in exon 28 were found in proband B.Homozygous insertional mutation g.82888_82889insCATG and compound heterozygous mutations g.94865G to A(Trp856X) and g.110698_110699delinsG probably have respectively induced type 3 von Willebrand diseases in the two probands.
- Published
- 2012
26. [Genotype and function analyses of four inherited dysfibrinogenemia pedigree caused by Arg16 amino acid substitution in fibrinogen Aα chain]
- Author
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Lin-lin, Jiang, Xue-feng, Wang, Qiu-lan, Ding, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Male ,Phenotype ,Genotype ,Fibrinogens, Abnormal ,Thrombin Time ,Fibrinogen ,Humans ,Female ,Blood Coagulation Tests ,Middle Aged ,Afibrinogenemia ,Pedigree - Abstract
To analyze the phenotype, genotype and function in four Chinese pedigrees with inherited dysfibrinogenemia.Routing tests including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), reptilase time (RT), the activities of antithrombin (AT), protein C (PC) and protein S (PS) were detected in four pedigrees. The activity and antigen of plasma fibrinogen were analyzed by Clauss and immunoturbidimetry methods, respectively. The molecular weight of fibrinogen of four probands was assessed by Western blot. The function of abnormal fibrinogen was evaluated by fibrinogen clottability, fibrinogen dynamic polymerization and fibrinolysis velocity, respectively. The sequences of all the exons and exon-intron boundaries of the three fibrinogen genes were amplified by PCR and analyzed by direct sequencing.Four probands had prolonged TT and RT, reduced plasma fibrinogen activity levels and normal antigen levels. The assays of Western blot showed no abnormal molecular weight of fibrinogen. Function tests revealed reduced fibrinogen clottability, delayed and decreased fibrinogen dynamic polymerization and reduced fibrinolysis velocity. Aα chain Arg16His and Arg16Cys mutations were identified in the four probands, respectively.The four probands with dysfibrinogenemia were caused by the mutations of Aα chain Arg16His or Arg16Cys. Mutation of the fibrinogen induced dysfunction of plasma fibrinogen.
- Published
- 2012
27. [Molecular mechanisms of recurrent venous thrombosis in two pedigrees with type I antithrombin deficiency]
- Author
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Yan, Xia, Qiu-lan, Ding, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Xue-feng, Wang, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Venous Thrombosis ,Fibrin ,Phenotype ,DNA Mutational Analysis ,Mutation ,Humans ,Female ,Middle Aged ,Pedigree - Abstract
To investigate the clinical phenotype, genotype and molecular mechanism of recurrent venous thrombosis in two Chinese pedigrees with type I antithrombin (AT) deficiency.The routine coagulation screening tests were detected, thrombin generation tests was performed to evaluate the hypercoagulation. Anticardiolipin antibody (ACA) and lupus anticoagulant (LA) were detected with enzyme-linked immunosorbent assay (ELISA) and diluted viper venom time assay (DVVT), respectively. The activities of protein C, protein S and AT (PC:A, PS:A, AT:A) were tested with chromogenic substrate assay or clotting method. The antigen of AT (AT:Ag) was performed with immunoturbidimetry methods. Western blot was used to analyze the molecular weight (MW) and the plasma levels of AT:Ag. All 7 exons and the flanking sequences were amplified by PCR. The mutation of AT gene and thrombophilia associated gene polymorphisms were analyzed by direct DNA sequencing. The expression plasmid of Ala404Asp mutant was constructed with site-directed mutagenesis method based on the wild-type (WT) AT cDNA contained in pcDNA 3.1 vector, and transiently expression of AT WT and the Ala404Asp mutant was performed using HEK293T cells. Cultured supernatant and cell lysates were collected and measured for AT:Ag by ELISA and Western blot.The results of routine coagulation tests in two probands were normal, thrombin generation tests indicated that proband 1 presented hypercoagulable state with 2.8 and 1.5 times higher of the endogenous thrombin potential (ETP) and peak height compared with that of normal, respectively. The levels of PC:A, PS:A, ACA and LA were normal. AT:A in proband 1 and proband 2 were 45% and 32%, and AT:Ag were almost half of the normal (121 mg/L and 158 mg/L), respectively. The results of Western blot showed that both probands' plasma levels of AT:Ag were lower than the normal pooled plasma and MW was normal. Two heterozygous mutations of g.3291C→T(Thr98Ile), g.13863CA(Ala404Asp) were identified in the probands, respectively. No proband had venous thrombosis associated gene polymorphisms. Expression in vitro showed that AT:Ag in culture media and lysates of Ala404Asp are 4.8% and 60.6% of that of WT, respectively.Thr98Ile and Ala404Asp mutation of AT gene significantly correlate with recurrent venous thrombosis in the two probands, respectively. Ala404Asp has not been described before. The mutant Ala404Asp protein can not be expressed due to impaired secretion and increased intracellular degradation, resulting in type I AT deficiency.
- Published
- 2012
28. [Molecular analysis of a patient with hemophilia A caused by FVIII His99Arg mutation]
- Author
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Huan-huan, Qin, Xue-feng, Wang, Qiu-lan, Ding, Ye-ling, Lu, Jing, Dai, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Male ,Factor VIII ,Genotype ,DNA Mutational Analysis ,Mutation, Missense ,Humans ,Hemophilia A - Abstract
To investigate the molecular mechanism of a Chinese hemophilia A patient in whom there was a discrepancy between the clinical bleeding symptoms and laboratory assay of FVIII activity (FVIII: C).FVIII: C was detected by chromogenic and one-stage methods, and FVIII: Ag by ELISA. The APTT corrected test was used to screen the FVIII inhibitor and PCR amplification to analyze all the exons and flanking sequences of F8 gene of the proband, PCR products were purified and sequenced directly. The corresponding gene sites of family members were detected according to the gene mutation sites. Two B domain deleted human FVIII mutant expression plasmids His99Arg and His99Ala (pRC/RS V - BDhFVIIIcDNA) were constructed and transfected into HEK293T transiently. FVIII: Ag and FVIII: C of the expression products were assayed.The proband APTT was prolonged, FVIII: Ag was 120% but FVIII: C1% and no FVIII inhibitor in plasma. The results of anticoagulation and fibrinolytic functions were normal. The cross reacting material positive (CRM+) hemophilia A was diagnosed. Gene analysis revealed a A28828G substitution in exon 3 resulted in a H (His) to R (Arg) missense mutation and the same heterozygous was identified in his mother. In vitro expression of FVIII: Ag and FVIII: C of His99Arg were 180.0% and 5.8% , respectively, while FVIII: Ag and FVIII: C of His99Ala were 45.0% and 20.0% of that of wild type, respectively. His99Arg and His99Ala were diagnosed as CRM+ and CRM- mutations, respectively.Both the two F VIII mutations could express FVIII protein. However, CRM His99Arg mutant protein has little FVIII procoagulant activity and His99Ala has reduced FVIII function by routine methods.
- Published
- 2012
29. [A single E726Q mutation in the membrane proximal α-helix of integrin β3 subunit induces membrane blebbing by disrupting the membrane-actin cortex interaction]
- Author
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Yong-Kui, Kong, Yue, Zhang, Ming-Hui, Lin, and Xiao-Dong, Xi
- Subjects
Cricetulus ,Cricetinae ,Mutation ,Integrin beta3 ,Mutagenesis, Site-Directed ,Animals ,CHO Cells ,Cell Surface Extensions ,Heterocyclic Compounds, 4 or More Rings ,Protein Structure, Tertiary - Abstract
The membrane proximal α helix of integrin β subunit cytoplasmic tails plays an important functional role by interacting with various intracellular proteins, namely talin, α-actinin or skelemin. This study was designed to investigate the functional role of 5 highly conserved charged amino acids (R(724), K(725), E(726), E(731), E(733)) within this α helix by site-directed mutagenesis. The result showed that CHO cells expressing the αIIbβ3E726Q mutant had the most prominent phenotype and characterized by defective cell spreading on immobilized fibrinogen. In addition, this E726Q mutation induced membrane blebbing in cells adherent on fibrinogen, and this blebbing could be inhibited by the myosin light chain ATPase inhibitor blebbistatin. It is concluded that the membrane proximal α-helix of integrin β3 subunit is important in linking the phospholipid membrane to the submembraneous actin cortex.
- Published
- 2011
30. ChemInform Abstract: An Atom-Efficient and Powerful Method for Direct Esterification of Silyl Ethers Catalyzed by HClO4-SiO2
- Author
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Qin-Pei Wu, Yi-Nan Shu, Ti-Jian Du, Yunzheng Li, Xiao-Dong Xi, Xi Chen, Qing-Shan Zhang, and Hai-Xia Liu
- Subjects
Silylation ,Chemistry ,Organic chemistry ,Atom (order theory) ,General Medicine ,Environmentally friendly ,Catalysis - Abstract
The fluoride-free one-pot method is environmentally friendly since the silyl protecting groups are transformed into acetates as the sole byproducts, which are readily recoverable and converted back to silyl chlorides as protecting agents by treatment with conc.
- Published
- 2011
31. [Analysis of phenotype and genotype in three Chinese pedigrees with inherited dysfibrinogenemia]
- Author
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Qi, Ouyang, Qiu-lan, Ding, Dan-dan, Huang, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Xue-feng, Wang, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Phenotype ,Asian People ,Base Sequence ,Genotype ,Mutation, Missense ,Fibrinogen ,Humans ,Female ,Middle Aged ,Afibrinogenemia ,Pedigree - Abstract
To analyze the phenotype and genotype in three Chinese pedigrees with inherited dysfibrinogenemia.Laboratory tests including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), reptilase time (RT), and the activities of antithrombin (AT:C), protein C (PC:C) and protein S(PS:C) were detected in three pedigrees. The activity and antigen of plasma fibrinogen (Fg) were analyzed by Clauss and immunoturbidimetry methods, respectively. The Fg of three probands was assessed by Western blot and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The sequences of all the exons and exon-intron boundaries of the three Fg genes FGA, GFB and FGG were amplified by PCR and analyzed by direct sequencing.Three probands had normal APTT, PT, PC:C, PS:C and AT:C, but prolonged TT and RT. The activity levels of the 3 probands's plasma Fg were reduced, but antigen levels were normal. Western blot and SDS-PAGE showed no abnormal molecular weight of Fg. The 3 heterozygous mutations of γ Arg275His, Aα Pro18Leu and Aα Arg16Cys were identified in the 3 probands, respectively.The three probands with dysfibrinogenemia were caused by the mutations of γ Arg275His, Aα Pro18Leu and Aα Arg16Cys, respectively. Both Aα Pro18Leu and Aα Arg16Cys were first reported in Chinese population.
- Published
- 2011
32. [Two novel mutations in one pedigree with hereditary Factor VII deficiency]
- Author
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Zhi-fang, Xing, Xue-feng, Wang, Jing, Dai, Ye-ling, Lu, Guan-qun, Xu, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Male ,Heterozygote ,Polymorphism, Genetic ,Base Sequence ,Child, Preschool ,Factor VII Deficiency ,DNA Mutational Analysis ,Mutation, Missense ,Humans ,Factor VII ,Pedigree ,Sequence Deletion - Abstract
To explore the mutations of coagulation factor VII (FVII) gene in one pedigree with hereditary FVII deficiency, and to investigate the molecular mechanisms of FVII deficiency.FVII gene mutations were analysed in the pedigree by direct DNA sequencing. The mutated DNA fragments were cloned into pMD19-T simple TA vector, and sequenced to confirm their distribution on chromosome. The plasma activity of FVII of the probands and their family members was detected with coagulation assay. The antigen of FVII were identified with ELISA.Three gene mutations were detected in the pedigree: A/G to C at 15386 resulting in Arg353Pro/Gln353Pro, A to T at 15274 resulting in Lys316Stop, all three mutations were heterozygotes. Three kinds of polymorphisms were identified in his father: A to G transition at position 15386 resulting in Arg353Gln, heterozygotic deletion of 2050 - 2059 cctatatcct in promoter and G to A mutation in intron 1a, the same polymorphisms were found in his grandfather. The three polymorphisms were located in the same chromosome of his father.Two mutations were found in the pedigree with hereditary FVII deficiency. One is nonsense mutation (Lys316Stop), the other is missense one (Gln353Pro). Gln353Pro and Lys316Stop might be the molecular mechanisms of FVII deficiency. The two novel mutations were reported for the first time in the literature.
- Published
- 2011
33. [Phenotype and genotype analysis of three Chinese pedigrees with von Willebrand disease]
- Author
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Huan-huan, Qin, Xue-feng, Wang, Qiu-lan, Ding, Guan-qun, Xu, Li-wei, Zhang, Jing, Dai, Ye-ling, Lu, Xiao-dong, Xi, and Hong-li, Wang
- Subjects
Adult ,Male ,Heterozygote ,von Willebrand Diseases ,Phenotype ,Genotype ,DNA Mutational Analysis ,von Willebrand Factor ,Humans ,Female ,Child ,Pedigree - Abstract
To analyze phenotype and genotype of three Chinese pedigrees with von Willebrand disease (vWD), and explore the molecular mechanism.Bleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor (vWF): ristocetin cofactor (RCof) (vWF:RCof), vWF antigen (vWF:Ag), vWF activity (vWF:A) test, vWF collagen binding assay (vWF:CB), vWF and Factor VIII (FVIII) binding assay (vWF:FVIII:B) and multimer analysis were used for phenotype diagnosis. Genomic DNA was extracted from the peripheral blood (PB). All the 52 exons and flanking sequences of the probands' vWF gene were amplified by PCR and analyzed by direct sequencing.APTT were prolonged in all three probands, while BT were normal excepting for proband 3. Plasma RIPA, vWF:RCo, vWF:Ag, vWF:A and vWF:CB were decreased in different extents. In multimer analysis, proband 3 lost the large and intermediate molecular weight multimers, while proband 1 and 2 were normal. Gene analysis in the three probands revealed three heterozygous missense mutations of 144067 G→A (R2287Q) in exon 39, 110374G→A (R1374H) and 110770C→T (S1506L) in exon 28 and heterozygous polymorphism 110667G→A (D1472H) in exon 28, respectively.The three heterozygous mutations (R2287Q, R1374H and S1506L) and an heterozygous polymorphism (D1472H) are genetic defects of the hereditary vWD of the three pedigrees respectively. R2287Q is a novel mutation reported for the first time in the literature.
- Published
- 2011
34. ChemInform Abstract: Synthesis and Antitumor Activity of Novel 2′,3′-Diethanethio-2′,3′,5′-trideoxy-5′-triazolonucleoside Analogues
- Author
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Jin-Lan Yu, Hongquan Yin, Xiao-Dong Xi, Qin-Pei Wu, Yan-Hong Liu, Yunzheng Li, Qing-Shan Zhang, and Ning-Ning Liu
- Subjects
Antitumor activity ,chemistry.chemical_compound ,chemistry ,Stereochemistry ,Nucleic acid ,General Medicine ,Derivative (chemistry) - Abstract
A series of novel triazoloribonucleosides (VII) is synthesized by the route described for the preparation of derivative (VI) and evaluated for antitumor activity.
- Published
- 2010
35. [Two new mutations of AT gene in type I inherited antithrombin deficiency.]
- Author
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Qiong, Chen, Ye-Ling, Lu, Guan-Qun, Xu, Qiu-Lan, Ding, Xue-Feng, Wang, Xiao-Dong, Xi, and Hong-Li, Wang
- Subjects
Heterozygote ,Antithrombin III Deficiency ,Phenotype ,Mutation ,Humans ,Pedigree - Abstract
To identify the clinical phenotype and gene mutation in two kindreds with type I inherited antithrombin (AT) deficiency.The coagulation and anticoagulation testing and thrombophilia screening were used for phenotypic diagnosis and immunonephelometry and chromogenic assay for plasma level of AT antigen (AT:Ag) and AT activity (AT:A), respectively. All of the seven exons and intron-exon boundaries and untranslation regions of AT gene were amplified by PCR, and the PCR products analysis was by direct sequencing. The corresponding gene sites of the two family members and healthy individuals were detected according to the gene mutation sites.The plasma levels of AT:Ag of proband 1 and proband 2 were 126 mg/L and 117 mg/L, and AT:A was 49% and 48%, respectively. Heterozygotic deletion of 3239-3240delCT in proband 1 and nonsense mutation 3206A--T (K70Stop) in proband 2 were rchaacterized in exon 2 of AT gene. And some of their family members were also detected with the heterozygotic gene mutation.Type I inherited antithrombin deficiency of the two probands were caused by AT gene mutation 3239-3240delCT and 3206A--T (K70Stop).
- Published
- 2010
36. [Analysis of phenotype and genotype in four Chinese pedigrees with inherited coagulation factor V deficiency.]
- Author
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Dan-Dan, Huang, Xue-Feng, Wang, Hua-Yun, Chen, Guan-Qun, Xu, Li-Wei, Zhang, Jin, Dai, Ye-Ling, Lu, Qiu-Lan, Ding, Xiao-Dong, Xi, and Hong-Li, Wang
- Subjects
Phenotype ,Genotype ,Factor V ,Humans ,Factor V Deficiency ,Pedigree - Abstract
To identify the phenotype and genotype in four Chinese pedigrees with inherited coagulation factor V (FV) deficiency.The tests of activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) were used for phenotype diagnosis. All the exons and exon-intron boundaries of F5 gene were amplified by PCR and analyzed by direct sequencing.The APTT and PT in each of the four probands were obviously prolonged, and both activity and antigen of FV in the four probands were extremely lower compared with that of normal mixed plasma. Sequencing of F5 gene in proband 1 identified a heterozygous mutation, G16088C (Asp68His), and four polymorphisms, T35788C (Met385Thr), A47295G (His1299Arg), A58668G (Met1736Val) and A74083G (Asp2194Gly), which were located in the same chromosome; proband 2 was homozygous for two mutations, C46253T (Arg952Cys) and C46724T(Gln1109stop); the F5 gene of proband 3 showed a homozygous missense mutation, C67793G(Pro2006Ala); and proband 4 was homozygous for one missense mutation, C74022T (Arg2174Cys).Five mutations (Asp68His, Arg952Cys, Gln1109stop, Pro2006Ala and Arg2174Cys) and four polymorphisms (Met385Thr, His1299Arg, Met1736Val and Asp2194Gly) may lead to type I inherited FV deficiency for these four probands, respectively. Gln1109stop, Pro2006Ala and Arg2174Cys haven't been identified before.
- Published
- 2010
37. [Preparation and characterization of a monoclonal antibody against human c-Kit]
- Author
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Hong-chen, Liu, Chao-ming, Mao, Xiao-yu, Su, Zheng, Ruan, Qiu-lan, Ding, Xue-feng, Wang, Hong-li, Wang, and Xiao-dong, Xi
- Subjects
Mice ,Mice, Inbred BALB C ,Proto-Oncogene Proteins c-kit ,Antibody Specificity ,Recombinant Fusion Proteins ,Blotting, Western ,Animals ,Antibodies, Monoclonal ,Humans ,Female ,Plasmids - Abstract
To prepare anti-c-Kit monoclonal antibodies and characterize their specificity of epitope recognition.cDNA encoding human c-Kit extracellular domain was constructed into a procaryotic expression vector pQE30 and the correctness of the reconstructed plasmid pQE30-KitD4-5 was verified by sequencing. The plasmid was transformed into E.coli M15 strain. Recombinant 6 x His pQE30-KitD4-5 was expressed after induction by IPTG for 4 h. Then SDS-PAGE results suggested that the products mainly formed inclusion bodies. The fusion protein was further purified with Ni-NTA-His affinity chromatography and then used to immunize BALB/c mice. The hybridomas were achieved by fusing the immunized spleen cells with the Sp2/0 myeloma cell line. The positive clones were screened by FCM with CHO-hKit cells. Hybidoma clones secreting anti-c-Kit antibodies were further subcloned and investigated for their biological activities by Western blot, rapid isotyping analysis and FCM.Recombinant human c-Kit fusion proteins were in vitro expressed and purified to be used as immunogen. One stable hybridoma cell line, which continuously secrets specific anti-c-Kit monoclonal antibody ((SRJ1)) was established. The biological activity studies showed that the monoclonal antibody recognized the natural c-Kit expressed on the Kasumi leukemia cell line, but failed to bind to the normal human peripheral blood cells. Interestingly, this monoclonal antibody failed to recognize a subpopulation of Kasumi cells that is reactive with the commercial anti-c-Kit mAb Ab81 suggesting that the c-Kit expressed by this subpopulation contains some sequencial and/or structural aberrations that are distinguishable by mAb SRJ1.With an immunization procedure using purified recombinant human c-Kit fusion proteins. a hybridoma cell line continuously and stably secreting anti-c-Kit monoclonal antibody has been established. The monoclonal antibody SRJ1 specifically recognizes human c-Kit expressed on the leukemia cells, and may provide a novel approach to analyze the possible structural variations of c-Kit expressed by different cells.
- Published
- 2009
38. [Study of signal transduction mediated by integrin alphaIIbbeta3 using a dominant negative model]
- Author
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Yuan-Jing, Lu, Xiao-Yu, Su, Zheng, Ruan, and Xiao-Dong, Xi
- Subjects
Cricetulus ,Cricetinae ,Recombinant Fusion Proteins ,Animals ,Receptors, Interleukin-2 ,CHO Cells ,Platelet Glycoprotein GPIIb-IIIa Complex ,Signal Transduction - Abstract
This study was purposed to investigate the role of integrin beta3 cytoplasmic domain in signal transduction mediated by integrin alphaIIbbeta3 and to explore the effect of integrin beta3 on signal transduction and specificity in condition without alphaIIb subunit. The fusion protein (Tac/beta3) was stably expressed in CHO cell line expressing GPIbIX, integrin alphaIIbbeta3 (IbIX/IIbIIIa-CHO cell line) by combining extracellular and transmembrane domains (Tac) of IL-2 receptor with integrin beta3 cytoplasmic domain (beta3) for formation of fusion protein (Tac/beta3). Then a series of tests were performed, including spreading and stable adhesion of IbIX/IIbIIIa-CHO cell line in solid phase fibrinogen (Fg), fibrin clot restriction and soluble fibrinogen binding, which represent outside-in and inside-out signal transduction events. The results showed that the bidirectional signal transduction mediated by alphaIIbbeta3 in IbIX/IIbIIIa-CHO/Tac-762 cells stably expressing Tac/beta3 was seriously inhibited. It is concluded that the Tac/beta3 can play a significant role in IbIX/IIbIIIa-CHO/Tac-762 cells through a dominant negative mode, the independent presence of beta3 subunit cytoplasmic domain can regulate the bidirectional signal transduction mediated by integrin alphaIIbbeta3.
- Published
- 2009
39. A mild, efficient, and selective procedure for transprotection of acetonides to acetates catalyzed with HClO4-SiO2
- Author
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Yi-Nan Shu, Qin-Pei Wu, Qing-Shan Zhang, Hai-Xia Liu, Ti-Jian Du, Xiao-Dong Xi, and Xi Chen
- Subjects
Anomer ,Perchlorates ,Chemistry ,Organic Chemistry ,General Medicine ,Acetates ,Silicon Dioxide ,Biochemistry ,Chemical synthesis ,Catalysis ,Analytical Chemistry ,Acetone ,Acetals ,Organic chemistry - Abstract
The transformation of acetonides into acetates is frequently required in synthetic chemistry. An efficient procedure for direct conversion of acetonides into acetates in the presence of HClO(4)-SiO(2) under mild conditions was developed. The acetonides of primary hydroxy groups are directly converted to diacetates, and the anomeric acetonides of furanosides are stereoselectively transformed into the corresponding acetyl beta-d-furanosides with a 2-acetoxyisopropyl group.
- Published
- 2009
40. [The application of thrombin generation tests to warfarin anticoagulation monitoring]
- Author
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Hua-Yun, Chen, Qiu-Lan, Ding, Li-Wei, Zhang, Guan-Qun, Xu, Jing, Dai, Xue-Feng, Wang, Xiao-Dong, Xi, and Hong-Li, Wang
- Subjects
Adult ,Aged, 80 and over ,Male ,Thrombin ,Anticoagulants ,Hemorrhage ,Middle Aged ,Atrial Fibrillation ,Prothrombin Time ,Humans ,Female ,International Normalized Ratio ,Warfarin ,Drug Monitoring ,Aged - Abstract
To explore the thrombin generation capacity in patients on warfarin therapy with different prothrombin time international normalized ratio (PT-INR), the capacity in relation to bleeding, and the application of thrombin generation tests to warfarin therapy monitoring.Seventy eight blood samples were taken from patients on warfarin therapy for more than 3 months owing to valve replacement or atrial fibrillation. The patients' case history and PT-INR were collected and thrombin generation tests were performed in all samples.Patients were ranked into three groups according to different PT-INR. There were 23 patients in group I with PT-INR from 1.51 to 2.00, 39 patients in group II with PT-INR from 2.01 to 3.00, and 16 patients in group III with PT-INR from 3.01 to 4.26. There were significant differences between each two of the three groups in lag time, peak, and ttpeak (time to peak) (P0.01). There was a significant difference between group I and group II in endogenous thrombin potential (ETP) (P = 0.0001), but not between group II and group III (P= 0.06). Five patients developed bleeding and their ETP was less than 15% of normal control.In patients on warfarin therapy, when the PT-INR was more than 3.0, increasing the dose of warfarin doesn' t decrease the thrombin generation, but increase bleeding risk. PT-INR combined with ETP may better reflect patient's coagulation status, therefore be of more significance in preventing bleeding.
- Published
- 2008
41. [Analysis of clinical features and genotype in three Chinese pedigrees with Glanzmann thrombasthenia]
- Author
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Pei-Pei, Jin, Wei-Zhang, Shen, Fang, Yang, Qiu-Lan, Ding, Xue-Feng, Wang, Xiao-Dong, Xi, and Hong-Li, Wang
- Subjects
Male ,Platelet Membrane Glycoprotein IIb ,Mutation ,Humans ,Female ,Exons ,Pedigree ,Thrombasthenia - Abstract
To study the clinical feature and alpha II b beta 3 gene mutations of three Glanzmann thrombasthenia (GT) pedigrees.Platelet counts (BPC), blood film, bleeding time, platelet aggregation and flow cytometry were used for phenotype diagnosis of all the patients. All the exons of alpha II b and beta 3 genes were amplified by polymerase chain reaction (PCR) and direct sequencing was performed for mutational screening. One hundred and three healthy blood donors were as normal controls.Three probands showed normal BPC, defective platelets aggregation, prolonged bleeding time and significantly reduced platelet aggregation to ADP, epinephrine, and collagen, while relatively normal aggregation to ristocetin. Flow cytometry showed platelet surface expressed alpha II b beta 3 was strongly reduced in proband 1 and proband 3 and mildly reduced in the amount of surface expressed alpha II b beta 3 (63%) in proband 2. Sequencing results showed that proband 1 had a G10A homozygous mutation in alpha II b, and a G1412T homozygous mutation in beta3. Compound heterozygous mutations in beta3, G1199A and 1525delC were identified in proband 2. No mutations in alpha II b beta 3 gene were identified in proband 3.Compound homozygous mutations, GI0A in alpha II b and G1412T in beta3, lead to GT in proband 1. Compound heterozygous mutations in beta3, G1199A and 1525delC, lead to GT in proband 2. The mutations of G10A, G1412T and 1525delC were reported for the first time in GT patients.
- Published
- 2008
42. Fibrinogen interaction of CHO cells expressing chimeric alphaIIb/alphavbeta3 integrin
- Author
-
Juan-juan, Chen, Xiao-yu, Su, Xiao-dong, Xi, Li-ping, Lin, Jian, Ding, and He, Lu
- Subjects
Cricetulus ,Cricetinae ,Mutant Chimeric Proteins ,Animals ,Fibrinogen ,Humans ,CHO Cells ,Integrin alphaVbeta3 ,Signal Transduction - Abstract
The molecular mechanisms of the affinity regulation of alphavbeta3 integrin are important in tumor development, wound repairing, and angiogenesis. It has been established that the cytoplasmic domains of alphavbeta3 integrin play an important role in integrin-ligand affinity regulation. However, the relationship of structure-function within these domains remains unclear.The extracellular and transmembrane domain of alphaIIb was fused to the alphav integrin cytoplasmic domain, and the chimeric alpha subunit was coexpressed in Chinese hamster ovary (CHO) cells with the wild-type beta3 subunit or with 3 mutant beta3 sequences bearing truncations at the positions of T741, Y747, and F754, respectively. The CHO cells expressing these recombinant integrins were tested for soluble fibrinogen binding and the cell adhesion and spreading on immobilized fibrinogen.All 4 types of integrins bound soluble fibrinogen in the absence of agonist stimulation, and only the cells expressing the chimeric alpha subunit with the wild-type beta3 subunit, but not those with truncated beta3, could adhere to and spread on immobilized fibrinogen.The substitution alphaIIb at the cytoplasmic domain with the alphav cytoplasmic sequence rendered the extracellular alphaIIbbeta3 a constitutively activated conformation for ligands without the need of pinside-outq signals. Our results also indicated that the COOH-terminal sequence of beta3 might play a key role in integrin alphaIIb/alphavbeta3-mediated cell adhesion and spreading on immobilized fibrinogen. The cells expressing alphaIIb/alphavbeta3 have enormous potential for facilitating drug screening for antagonists either to alphavbeta3 intracellular interactions or to alphaIIbbeta3 receptor functions.
- Published
- 2008
43. A coma-free super-high resolution optical spectrometer using 44 high dispersion sub-gratings
- Author
-
Hua-Tian Tu, An-Qing Jiang, Jian-Ke Chen, Wei-Jie Lu, Kai-Yan Zang, Hao-Qi Tang, Osamu Yoshie, Xiao-Dong Xiang, Young-Pak Lee, Hai-Bin Zhao, Yu-Xiang Zheng, Song-You Wang, Junpeng Guo, Rong-Jun Zhang, Jing Li, Yue-Mei Yang, W. D. Lynch, and Liang-Yao Chen
- Subjects
Medicine ,Science - Abstract
Abstract Unlike the single grating Czerny–Turner configuration spectrometers, a super-high spectral resolution optical spectrometer with zero coma aberration is first experimentally demonstrated by using a compound integrated diffraction grating module consisting of 44 high dispersion sub-gratings and a two-dimensional backside-illuminated charge-coupled device array photodetector. The demonstrated super-high resolution spectrometer gives 0.005 nm (5 pm) spectral resolution in ultra-violet range and 0.01 nm spectral resolution in the visible range, as well as a uniform efficiency of diffraction in a broad 200 nm to 1000 nm wavelength region. Our new zero-off-axis spectrometer configuration has the unique merit that enables it to be used for a wide range of spectral sensing and measurement applications.
- Published
- 2021
- Full Text
- View/download PDF
44. [Establishment of urokinase receptor gene antisense RNA transfer system and its application in leukemia research]
- Author
-
Xia, Bai, Jian-Xin, Fu, Xiao-Dong, Xi, and Chang-Geng, Ruan
- Subjects
Leukemia ,Retroviridae ,Gene Transfer, Horizontal ,Matrix Metalloproteinase 9 ,Humans ,RNA, Antisense ,Receptors, Cell Surface ,U937 Cells ,Flow Cytometry ,Receptors, Urokinase Plasminogen Activator - Abstract
Overexpression of urokinase-type plasminogen activator receptor (uPAR) on tumor cell surface is essential for invasion and metastasis in a variety of tumor cells. To establish a retroviral-mediated antisense RNA transfer system of uPAR gene for exploring its function on down-regulation of uPAR expression in leukemia cells, the retroviral vector LaCD87SN was constructed by inserting uPAR gene into LXSN vector in an antisense orientation. An uPAR gene antisense RNA transfer system was established by liposome-mediated transfection in combination with cross infection with retrovirus. Human leukemia cells U937 were transduced with aCD87 amphotropic retrovirus, expressing uPAR antisense RNA, and the U937/aCD87 cells was obtained by G418 selection. The integration and expression of antisense uPAR gene were analyzed by polymerase chain reaction (PCR) assay. The cell surface expression of CD87 and the activities of matrix metalloproteinase (MMP) were assayed by flow cytometry (FCM) and gelatin zymography, respectively. The results showed that the amphotropic retroviral producers Am12/aCD87, which expressed antisense RNA of uPAR gene with a titer of 6.3 x 10(5) cfu/ml in supernatants, were obtained by means of transfection and superinfection. U937/aCD87 cells were established by continuative G418 selection after retrovirus infection. In U937/aCD87 cells, the integrated provirus and the overexpression of antisense uPAR gene was confirmed. Compared with U937/NeoR cells, FCM analysis revealed that CD87 expression on U937/aCD87 cell surface was not downregulated significantly. However, MMP-9 secretion was significantly suppressed in U937/aCD87 cells. In conclusion, although the retroviral-mediated antisense RNA transfer could not efficiently suppress uPAR expression on leukemic cell surface, it may interfere the uPAR-MMP interactions.
- Published
- 2004
45. Clinical significance of day 5 peripheral blast clearance rate in the evaluation of early treatment response and prognosis of patients with acute myeloid leukemia.
- Author
-
Cong Yu, Qing-lei Kong, Yun-xiang Zhang, Xiang-qin Weng, Jing Wu, Yan Sheng, Chun-lei Jiang, Yong-mei Zhu, Qi Cao, Shu-min Xiong, Jun-min Li, Xiao-dong Xi, Sai-juan Chen, and Bing Chen
- Subjects
ACUTE myeloid leukemia ,CANCER chemotherapy ,RECEIVER operating characteristic curves ,FLOW cytometry ,CANCER patients - Abstract
Background: Minimal residual disease detection in the bone marrow is usually performed in patients with acute myeloid leukemia undergoing one course of induction chemotherapy. To optimize the chemotherapy strategies, more practical and sensitive markers are needed to monitor the early treatment response during induction. For instance, peripheral blood (PB) blast clearance rate may be considered as such a monitoring marker. Methods: PB blasts were monitored through multiparameter flow cytometry (MFC). Absolute counts were determined before treatment (D
0 ) and at specified time points of induction chemotherapy (D3 , D5 , D7 , and D9 ). The cut-off value of D5 peripheral blast clearance rate (D5 -PBCR) was defined through receiver operating characteristic (ROC) analysis. Prognostic effects were compared among different patient groups according to D5 -PBCR cut-off value. Results: D5 -PBCR cut-off value was determined as 99.55%. Prognostic analysis showed that patients with D5 -PBCR ≥99.55% more likely achieved complete remission (94.6% vs. 56.1%, P < 0.001) and maintained a relapse-free status than other patients (80.56% vs. 57.14%, P = 0.027). Survival analysis revealed that relapse-free survival (RFS) and overall survival (OS) were longer in patients with D5 -PBCR ≥99.55% than in other patients (two-year OS: 71.0% vs. 38.7%, P = 0.011; two-year RFS: 69.4% vs. 30.7%, P = 0.026). In cytogenetic-molecular intermediate-risk group, a subgroup with worse outcome could be distinguished on the basis of D5 -PBCR (<99.55%; OS: P = 0.033, RFS: P = 0.086). Conclusions: An effective evaluation method of early treatment response was established by monitoring PB blasts through MFC. D5 -PBCR cut-off value (99.55%) can be a reliable reference to predict treatment response and outcome in early stages of chemotherapy. The proposed marker may be used in induction regimen modification and help optimize cytogenetic-molecular prognostic risk stratification. [ABSTRACT FROM AUTHOR]- Published
- 2015
- Full Text
- View/download PDF
46. N-trans-Feruloyloctopamine Wakes Up BBC3, DDIT3, CDKN1A, and NOXA Signals to Accelerate HCC Cell Apoptosis
- Author
-
Bin Ma, Jing Li, Wen-Ke Yang, Mei-Gui Zhang, Xiao-Dong Xie, and Zhong-Tian Bai
- Subjects
Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 ,Cytology ,QH573-671 - Abstract
N-trans-Feruloyloctopamine (FO), a natural compound, was reported in our previous study to inhibit a tumor cell malignant phenotype by AKT- and EMT-related signals and might be used as a promising drug for HCC treatment. However, the specific targets and detailed mechanisms still need to be clarified. Screening with RNA-Seq in Huh7 cells treated with FO revealed that 317 genes were modulated, of which 188 genes were upregulated and 129 genes were downregulated. Real-time cell analyzer and flow cytometry data reveal that tumor cell proliferation and apoptosis were impacted by FO. DAVID bioinformatic data showed that most of the biological process GO terms are related to proliferation and apoptosis. KEGG enrichment analysis showed that FO mainly regulates PI3K-AKT- and apoptosis-related signals, in which BBC3, DDIT3, NOXA, and CDKN1A on the surface serve as the novel targets of FO inducing HCC cell apoptosis. The result implied that FO might exacerbate HCC cell apoptosis by regulating BBC3, DDIT3, CDKN1A, and NOXA signals. The obstacle effect of FO can provide new targets and new credibility for the treatment of liver cancer.
- Published
- 2021
- Full Text
- View/download PDF
47. Separation, Purification, Structure Analysis, In Vitro Antioxidant Activity and circRNA-miRNA-mRNA Regulatory Network on PRV-Infected RAW264.7 Cells of a Polysaccharide Derived from Arthrospira platensis
- Author
-
Mi-Xia Cao, Xiao-Dong Xie, Xin-Rui Wang, Wen-Yue Hu, Yi Zhao, Qi Chen, Lu Ji, Ying-Yi Wei, Mei-Ling Yu, and Ting-Jun Hu
- Subjects
Arthrospira platensis polysaccharide ,pseudorabies virus ,antioxidant ,structure ,circRNA-miRNA-mRNA ,Therapeutics. Pharmacology ,RM1-950 - Abstract
To investigate the structure of Arthrospira platensis polysaccharide (PAP) (intracellular polysaccharide) and the antioxidant activity of the first component of PAP (PAP-1) on pseudorabies virus (PRV) -infected RAW264.7 cells. The PAP was separated and purified by the Cellulose DE-52 chromatography column and Sephacryl S-200 high-resolution gel column to obtain PAP-1. The antioxidant activity and regulation of PAP-1 on PRV-infected RAW264.7 cells of circRNA-miRNA-mRNA network were investigated by chemical kit, Q-PCR, and ce-RNA seq. The results indicated that the molecular weight (Mw) of PAP-1, which was mainly composed of glucose and eight other monosaccharides, was 1.48 × 106 Da. The main glycosidic bond structure of PAP-1 was →4)-α-D-Glcp-(1→. PAP-1 may be increased the antioxidant capacity by regulating the circRNA-miRNA-mRNA network in PRV-infected RAW264.7 cells. This study provided a scientific foundation for further exploring the antioxidant activity of PAP-1 based on its structure.
- Published
- 2021
- Full Text
- View/download PDF
48. Preferential eradication of acute myelogenous leukemia stem cells by fenretinide.
- Author
-
Hui Zhang, Jian-Qing Mi, Hai Fang, Zhao Wang, Chun Wang, Lin Wu, Bin Zhang, Minden, Mark, Wen-Tao Yang, Huan-Wei Wang, Jun-Min Li, Xiao-Dong Xi, Sai-Juan Chen, Ji Zhang, Zhu Chen, and Kan-Kan Wang
- Subjects
ACUTE myeloid leukemia treatment ,STEM cells ,FENRETINIDE ,VITAMIN A ,METHYLCELLULOSE ,XENOGRAFTS - Abstract
Leukemia stem cells (LSCs) play important roles in leukemia initiation, progression, and relapse, and thus represent a critical target for therapeutic intervention. However, relatively few agents have been shown to target LSCs, slowing progress in the treatment of acute myelogenous leukemia (AML). Based on in vitro and in vivo evidence, we report here that fenretinide, a welltolerated vitamin A derivative, is capable of eradicating LSCs but not normal hematopoietic progenitor/stem cells at physiologically achievable concentrations. Fenretinide exerted a selective cytotoxic effect on primary AML CD34
+ cells, especially the LSC-enriched CD34+ CD38- subpopulation, whereas no significant effect was observed on normal counterparts. Methylcellulose colony formation assays further showed that fenretinide significantly suppressed the formation of colonies derived from AML CD34+ cells but not those from normal CD34+ cells. Moreover, fenretinide significantly reduced the in vivo engraftment of AML stem cells but not normal hematopoietic stem cells in a nonobese diabetic/ SCID mouse xenotransplantation model. Mechanistic studies revealed that fenretinide-induced cell death was linked to a series of characteristic events, including the rapid generation of reactive oxygen species, induction of genes associated with stress responses and apoptosis, and repression of genes involved in NF-κB and Wnt signaling. Further bioinformatic analysis revealed that the fenretinide-down-regulated genes were significantly correlated with the existing poor-prognosis signatures in AML patients. Based on these findings, we propose that fenretinide is a potent agent that selectively targets LSCs, and may be of value in the treatment of AML. [ABSTRACT FROM AUTHOR]- Published
- 2013
- Full Text
- View/download PDF
49. Structural framework of c-Src activation by integrin β3.
- Author
-
Run Xiao, Xiao-Dong Xi, Zhu Chen, Sai-Juan Chen, and Guoyu Meng
- Subjects
- *
INTEGRINS , *CELL adhesion molecules , *GLYCOPROTEINS , *CANCER cells , *CELLS - Abstract
The integrin β3-mediated c-Src priming and activation, via the SH3 domain, is consistently associated with diseases, such as the formation of thrombosis and the migration of tumor cells. Conventionally, activation of c-Src is often induced by the binding of proline-rich sequences to its SH3 domain. Instead, integrin β3 uses R760GT762 for priming and activation. Because of the lack of structural information, it is not clear where RGT will bind to SH3, and under what mechanism this interaction can prime/activate c-Src. In this study, we present a 2.0-Å x-ray crystal structure in which SH3 is complexed with the RGT peptide. The binding site lies in the "N"-Src loop of the SH3 domain. Structure-based site-directed mutagenesis showed that perturbation on the "N"-Src loop disrupts the interaction between the SH3 domain and the RGT peptide. Furthermore, the simulated c-Src:β3 complex based on the crystal structure of SH3:RGT suggests that the binding of the RGT peptide might disrupt the intramolecular interaction between the SH3 and linker domains, leading to the disengagement of Trp260:"C"-helix and further activation of c-Src. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
50. Effect of statins on blood pressure: Analysis on adverse events released by FDA
- Author
-
Tao You, Xing-guang Liu, Xiao-dong Hou, Xin-kuan Wang, Han-hui Xie, Fan Ding, Kang Yi, Peng Zhang, and Xiao-dong Xie
- Subjects
atorvastatin ,blood pressure ,fda adverse event reporting system ,hypotension ,simvastatin ,rosuvastatin ,Diseases of the circulatory (Cardiovascular) system ,RC666-701 - Abstract
Objective: As a class of cholesterol-lowering drugs, statins have been reported to cause unexpected decrease in blood pressure (BP). However, most studies in this issue were subject to inadequate study design or very small sample size. The present study was designed to examine the BP-lowering effect of various statins. Methods: Here we retrieved 5.9 million clinical reports submitted to FDA Adverse Event Reporting System (FAERS) from 2004 to 2015. Meta-analysis was performed to estimate the overall reporting odds ratio (ROR) of hypotension adverse events concurrent with various statins (i.e., atorvastatin, simvastatin, and rosuvastatin). Results: Comparing the reporting rate of hypotension event between statins and other drugs found that atorvastatin (pooled ROR = 1.26, adjusted p-value = 8.60 × 10−4) and simvastatin (pooled ROR = 1.94, adjusted p-value = 4.16 × 10−45) were significantly associated with reduction in BP. On the other hand, the association between rosuvastatin and hypotension was observed to be nonsignificant (adjusted p-value = 0.65). Conclusion: To our knowledge, this is the first pooled analysis on large-scale data of adverse events to identify the BP-lowering effect of statins. The results will contribute to the development of novel statin-based antihypertensive therapies. In addition, the differential effects of individual statins can warrant subsequent research on the underlying mechanisms of BP control.
- Published
- 2017
- Full Text
- View/download PDF
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