1.Recombinant expression and characterization of CD2-binding domain of Macaca mulatta lymphocyte function-associated antigen 3 in Pichia pastoris.
Jian ZHU ; Shengyun ZHU ; Hao YANG ; Xiaofeng LU ; Lin WAN
Journal of Biomedical Engineering 2015;32(1):120-125
Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.
Animals
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CD58 Antigens
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biosynthesis
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Humans
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Immunoglobulin G
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Lymphocyte Activation
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Macaca mulatta
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Pichia
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Plasmids
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Protein Interaction Domains and Motifs
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Recombinant Fusion Proteins
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biosynthesis
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T-Lymphocytes
2.Analysis of the T cell subsets in the workers exposed to lead.
Lin WANG ; Xiaofeng LI ; Ruxue ZHANG ; Fang WAN ; Lin ZHAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2014;32(6):435-437
Adult
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Female
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Humans
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Lead
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Male
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Middle Aged
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Occupational Exposure
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T-Lymphocyte Subsets
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immunology
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Young Adult
3.Molecular Cloning, Recombinant Expression and Functional Characterization of the Soluble Tumor Necrosis Factor-related Apoptosis-inducing Ligand for the Macaca mulatta.
Fan MIAOMIAO ; Dianlong JIA ; Hao YANG ; Lin WAN ; Xiaofeng LU
Journal of Biomedical Engineering 2015;32(3):605-611
Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropyl β-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
Animals
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Antineoplastic Agents
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Apoptosis
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Cell Line, Tumor
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Cloning, Molecular
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Humans
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Leukocytes, Mononuclear
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Macaca mulatta
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Mice
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Plasmids
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TNF-Related Apoptosis-Inducing Ligand
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genetics
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metabolism
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Xenograft Model Antitumor Assays
4.Development of WMZ-03 anaesthesia workstation with chest intergrated with apparatus
Taihu WU ; Zhenxing SONG ; Zhen WAN ; Xiaofeng ZHANG
Chinese Medical Equipment Journal 1989;0(03):-
This paper introduces a type of anaesthesia workstation which is composed of two vertical medical chests, i.e. the upper and lower parts. Taken out of the chest, the two parts are butted against and locked to make the workstation ready for operation. After operation is finished, the two parts are uncoupled and put into the chests, and then the workstation can be stored and transported with the chests closed.
5.The diagnosis of orbital tumor with dynamic contrast-enhanced MRI
Xiaofeng TAO ; Weiping WAN ; Xiangsheng XIAO ; Zengru SHI
Chinese Journal of Radiology 2001;0(04):-
Objective To improve the diagnostic value of MRI on orbital diseases,especially on benign and malignant tumors,through analyzing the characteristics of orbital diseases with dynamic contrast-enhanced MRI.Methods Forty-one patients with solitary orbital masses, including 31 benign tumors and 10 malignant tumors,underwent MR examinations. All the cases were verified by pathology.1.5 T MR system and head coil were employed.Conventional scan was performed with axial,sagittal,and coronal T_1WI,as well as axial T_2WI and the slice thickness of 4 mm.After the injection of contrast agents,dynamic contrast-enhanced MRI was immediately performed so as to observe dynamic changes of lesions.Enhancement rates(E%) was calculated and time-intensity curve was portraited according to the signal intensity of conventional scan and dynamic-contrast enhanced scan.SPASS Ver10.0 was used to do the statistical analysis,while cartographies were completed with Powerpoint.Results(1)Comparision between benign and malignant tumors in average enhanced rates of the 1~(st) minute: it was higher than 60% in malignant tumors,while lower than 60% in benign ones(P
6.Application of ISSR-PCR to identification of different Dendrobium Sw. species
Ying SHEN ; Cheng XU ; Xiaofeng WAN ; Ming ZHANG
Chinese Traditional and Herbal Drugs 1994;0(03):-
Objective To identify nine species of Dendrobium Sw. by inter-simple sequence repeats-polymerase chain reaction (ISSR-PCR) method and to investigate the difference among the species of Dendrobium Sw. at DNA molecular level. Methods Ten primers constituted by simple sequence repeats (SSR) were tested for PCR and sepharose electrophoresis of the nine species. Results Seven of the ten primers amplified polymorphic bands. The number of polymorphic band positions of each primer ranged from 7 to 14, and the size of fragments ranged from 220 to 1 260 bp. The amplification patterns of two primers, namely UBC-807 and UBC-864, were higher in terms of polymorphic and amplified band ratio. Each of them was able to distinguish all the examined species. Conclusion ISSR-PCR provides a quick, reliable molecular marker technique for identification of different species of Dendrobium Sw.
7.Impact thermal environment of armored vehicle cabin on humans
Yanan WU ; Haijun JIA ; Wei ZHANG ; Xiaofeng LIU ; Xueyou WANG ; Weili CHAI ; Yi LI ; Hong WAN
Military Medical Sciences 2017;41(1):12-16
Objective To prolong the working time of the crew under a high temperature and high humidity environment in armored vehicles , improve work efficiency , and to prevent excessive heat load from doing any harm to the human body . Methods The man-machine environment was analyzed , the corresponding environment and the conditions aboard were further quantitatively studied , and man-machine-environment interactions were analyzed .Also,a questionnaire survey was conducted at the same time .Results and Conclusion The thermal equilibrium and thermal comfort calculation program were compiled and validated through the questionnaire survey in order to provide support for environment assessment and protection design .
8.Relationship between psychological quality and performance capacity among armored crew
Weili CHAI ; Shuwen GUO ; Xueyou WANG ; Haiming WU ; Hong WAN ; Xiaofeng LIU ; Yanan WU
Military Medical Sciences 2016;40(2):110-112
Objective To investigate the relationship between psychological quality and performance capacity of the crew in armored equipment so as to provide scientific evidence for the training of crew .Methods The psychological quality of 82 crew members of armored cavalry was tested and analyzed by neurobehavioral testers and questionnaires .Results and Conclusion The education level and age had no significant effect on the performance capacity among the armored crew . The test results of psychological quality parameters showed the same trend as the performance capacity of the crew , but their psychological health status had obvious effect on the performance capacity .The psychological quality of the crew had great influence on their performance capacity ,so we should pay attention to mental intervention ,enhance psychological protection ability of the crew , and improve their combat effectiveness continuously .
9.Clinical observation of cinobufacini injection used to treat moderate and advanced primary liver cancer
Zhe CHEN ; Xiaofeng ZHAI ; Yonghua SU ; Xuying WAN ; Jin LI ; Juemin XIE ; Bo GAO
Journal of Integrative Medicine 2003;1(3):184-6
OBJECTIVE: To observe the clinical effect of cinobufacini injection in treating moderate and advanced primary liver cancer (PLC). METHODS: One hundred patients with moderate and advanced PLC were randomly divided into cino-treated group (50 patients) and control group (50 patients). The quality of life, tumor size, some changes of laboratory tests, and survival time were observed. RESULTS: The progressive rate of cino-treated group (18%) was lower than that of the control group (32%). The quality of life of the cino-treated group (80%) was better than that of the control group (72%), but without statistical significance. The survival rate of >12 months of the cino-treated group (30%) was higher than that of the control group (18%). The patients' liver function such as serum total bilirubin and ALT decreased obviously in the cino-treated group while increased a lot in the control group. The level of AFP increased after treatment with statistical significance in the control group while there was no statistical significance in the cino-treated group. CONCLUSION: Cinobufacini injection can not only inhibit the proliferation of cancer, but also protect liver function, improve quality of life and prolong survival time.
10.Effects of multimodal combination dialysis on Klotho protein, FGF-23 and BNP in patients with maintenance hemodialysis
Xiaofeng DENG ; Qing DAI ; Li WAN ; Ling TANG ; Yue SHU ; Hengfen XIAO ; Yuanyuan BI ; Hongfen YANG
Chinese Critical Care Medicine 2017;29(7):636-639
Objective To discuss the effects of multimodal combination dialysis on Klotho protein, fibroblast growth factor-23 (FGF-23) and brain natriuretic peptide (BNP) in patients with maintenance hemodialysis (MHD). Methods A randomized controlled trial (RCT) was conducted. 120 patients who was diagnosed with chronic renal failure (CRF) uremia receiving MHD over 3 months admitted to Blood Purification Centre of Department of Nephrology of the Second People's Hospital of Guiyang from December 2015 to December 2016 were enrolled, who were randomly divided into hemodialysis (HD) group (HD for 8 times a month), HD + hemofiltration (HF) group (HD for 8 times a month + HF once a month), and HD + HF + hemoperfusion (HP) group (HD for 8 times a month + HF for 4 times a month + HP once a month), with 40 patients in each group. Before and after treatment for 6 months and 12 months, blood was taken from venous circuit tube, the serum Klotho protein and FGF-23 levels were determined by enzyme linked immunosorbent assay (ELISA), and the serum BNP level was determined by electrochemiluminescence. Results 120 patients with MHD were enrolled in the final analysis without withdrawal. There were no significant differences in the levels of Klotho protein, FGF-23, or BNP before enrollment among the three groups (all P > 0.05). Compared with those before enrollment, the levels of serum Klotho protein after enrollment in three groups showed a sustained upward tendency, which were higher in HD + HF + HP group than in HD + HF group and HD group (μg/L: 2.59±0.61, 1.63±0.35, 1.13±0.26 at 6 months, F = 119.374, P = 0.000; 6.98±1.21, 3.57±1.03, 2.12±0.43 at 12 months, F = 275.675, P = 0.000); the levels of FGF-23 showed a sustained downward tendency, which were lower in HD + HF + HP group than in HD + HF group and HD group (ng/L: 69.22±38.26, 132.28±61.18, 178.50±74.64 at 6 months, F = 33.509, P = 0.000; 32.81±17.32, 87.93±43.27, 146.33±69.28 at 12 months, F = 55.466, P = 0.000);the BNP showed a similar tendency as FGF-23 (ng/L: 4083.39±2864.53, 7245.69±4643.81, 7969.12±5360.85 at 6 months, F = 8.758, P = 0.000; 1521.86±894.63, 4554.32±1969.84, 5013.89±2033.64 at 12 months, F = 49.003, P = 0.000). Conclusion Multimodal combination dialysis can increase the Klotho protein level, and decrease the levels of FGF-23 and BNP in MHD patients with CRF uremia.