1.Expression of Truncated NSP2 Protein of Porcine Reproduction and Respiratory Syndrome Virus in E.coli and Preparation of Monoclonal Antibodies Against NSP2 Protein
Hai-Yan WANG ; Ping JIANG ; Yi-Jun DU ; Yu-Feng LI ; Jun-Xing LI ; Fang SHEN ;
China Biotechnology 2006;0(02):-
The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.
2.Expression of NSP 3AB Gene of Encephalomyocarditis Virus(EMCV)in E.coli and Development of Monoclonal Antibodies Against 3AB Protein
Fang SHEN ; Ping JIANG ; Yu-Feng LI ; Jun-Xing LI ; Hai-Yan WANG ;
China Biotechnology 2006;0(09):-
Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.
3.Modified Shengma Biejia Decoction Combined with CAG Program for Elderly Acute Myeloid Leuke- mia Patients with Yin Deficiency Toxin Stasis Syndrome.
Xing-bin DAI ; Xue-mei SUN ; Peng-jun JIANG ; Hai-wen NI ; Jian-yi CHEN ; Wen-xi ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(2):149-154
OBJECTIVETo observe the efficacy and safety of modified Shengma Biejia Decoction (MSBD) combined with CAG program in treating elderly acute myeloid leukemia (AML) patients with yin deficiency toxin stasis syndrome (YDTSS).
METHODSTotally 46 elderly AML patients were assigned to the treatment group (24 cases; treated with MSBD + CAG) and the control group (22 cases; treated with CAG + placebos of Chinese medicine) according to random digit table. The therapeutic course of CM placebo or MSBD was 21 days. The clinical efficacy and adverse reactions were observed. Meanwhile, physical state (ECOG Score), transfusion dependency, and TCM syndrome score were compared before and after treatment.
RESULTS(1) The complete remission rate was 54% (13/24) and the objective response rate (ORR) was 71% (17/24) in the treatment group, obviously higher than those of the control group [36% (8/22); 54% (13/24)], with statistical difference (P = 0.036, 0.042). When comparing the efficacy based on risk level, the moderate and poor ORR was 71% (10/14) and 67% (6/9) in the treatment group, and 57% (8/14) and 33% (2/6) in the control group, with statistical difference between the two groups (P = 0.048; P = 0.010). (2) Compared with before treatment in the same group, the ECOG score significantly decreased, the average infusion time of red cells and platelets were markedly prolonged in the treatment group after treatment (P < 0.05). ECOG score, the average infusion time of red cells and platelets were significantly better in the treatment group than in the control group after treatment (P < 0.05). (3) Compared with before treatment in the same group, scores of fever, hemorrhage, and bone pain were markedly reduced in the control group (P < 0.05); scores of fever, fatigue, hemorrhage, dry mouth, and bone pain were markedly reduced in the treatment group (P < 0.05). Better effect in relief of fever, fatigue, hemorrhage, dry mouth, and so on was obtained in the treatment group than in the control group (P < 0.05). (4) In aspect of hematotoxicity, the incidence of neutropenia, anemia, thrombocytopenia was obviously lower in the treatment group than in the control group [29.2% (7/24) vs 54.5% (12/22); 16.7% (4/ 24) vs 45.5% (10/22); 33.3% (8/24) vs 63.6% (14/22); P < 0.05]. The incidence of fatigue and anorexia was obviously lower in the treatment group than in the control group [37.5% (9/24) vs 63.6% (14/22), 37.5% (9/24) vs 81.8% (18/22); P < 0.05].
CONCLUSIONMSBD combined with CAG program in treating elderly AML patients with YDTSS, with efficacy enhancing toxicity reducing effect, had distinct advantages in improving physical condition and clinical symptoms, and reducing transfusion dependency.
Aclarubicin ; therapeutic use ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cytarabine ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Medicine, Chinese Traditional ; Phytotherapy ; Yin Deficiency ; drug therapy
5.Dose-effect relationship between premature chromosome and irradiation dose.
Bo JIANG ; Qiang LIU ; En-Hai JIANG ; Zhi-Wei XING ; Xin-Ran ZHAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(12):714-717
OBJECTIVETo explore the dose-effect relationship between premature chromosome condensation induced by Calyculin A and irradiation dose.
METHODSThe human peripheral blood was irradiated by (137)Cs gamma radial. The irradiation dose included 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 Gy. The premature chromosome condensation induced by Calyculin A was observed, and dyed by centromeric banding.
RESULTSThere was the quadratic relation between the total aberration, fragment, dicentric+centric ring (dic+r) ration and irradiation dose.
CONCLUSIONPremature chromosome condensation induced by Calyculin A can be used as a biodosimetry.
Cell Line ; Chromosome Aberrations ; radiation effects ; Chromosome Banding ; Chromosomes ; radiation effects ; Dose-Response Relationship, Radiation ; Female ; Gamma Rays ; adverse effects ; Humans ; Lymphocytes ; radiation effects ; Male ; Oxazoles ; pharmacology ; Young Adult
6.Sectional anatomical analysis of auricular and middle ear malformation in patients with microtia.
Juan HAN ; Jiu-xing LU ; Ning XING ; Lin LIN ; Hai-yue JIANG ; Hong-xing ZHUANG
Chinese Journal of Plastic Surgery 2011;27(3):217-221
OBJECTIVETo analyze the sectional anatomical features of auricular and middle ear malformation in patients with microtia so as to improve the clinical classification and the instruction of surgery.
METHODSFrom Jun. to Dec. 2009, 36 cases with microtia were selected in the center of auricular reconstruction in Plastic Surgery Hospital, including 22 cases of unilateral microtia and 14 cases of bilateral microtia. 22 patients with unilateral microtia were studied with the contralateral healthy ears as controls. Spiral CT was performed for high-resolution scan of the temporal bone. The coronal, sagittal and 3D reconstruction images were created with Mimic software. Several distances and degrees were measured.
RESULTSThe patients were classified by Max classification. The anteroposterior diameter and the vertical diameter of tympanic cavity were (7.75 +/- 1.92) mm and (14.66 +/- 4.75) mm for type I; (6.17 +/- 2.56) mm and(14.35 +/- 5.12) mm for type II; (6.31 +/- 3.40) mm and (9.97 +/- 4.36) mm for type III (P = 0.001). The mastoid pneumatization degree for type I, II, III were 13.33%, 13.64%, 30.77% in sclerotic type, 13.33%, 18.18%, 7.69% in diploe type, 0, 9.09%, 38.46% in composite type, 73.33%, 59.09%, 23.08% in pneumatic type (chi2 = 24.11, P = 0.002). The cover of fenestra vestibuli by facial nerve was 21.43%, 47.62%, 54.55% (chi2 = 23.44, P = 0.002) for type I, II, III. There was a statistical difference between the microtia group and the control group.
CONCLUSIONSAccording to the Max classification, the middle ear malformation changed along the auricular malformation. The anatomical variations was complicated in type II microtia, which should be sub-classified.
Adolescent ; Adult ; Anatomy, Cross-Sectional ; Child ; Child, Preschool ; Ear ; abnormalities ; diagnostic imaging ; Ear, External ; abnormalities ; diagnostic imaging ; Ear, Middle ; abnormalities ; diagnostic imaging ; Female ; Humans ; Male ; Tomography, Spiral Computed ; Young Adult
7.Effect of trichloroethylene intake via drinking water on Th17 cells in BALB/c mice.
Tong SHEN ; Jin WANG ; Hui XU ; Shu-Hai XU ; Tao JIANG ; Qi-Xing ZHU
Chinese Journal of Preventive Medicine 2012;46(2):152-157
OBJECTIVEThis study aimed to investigate the effect of trichloroethylene (TCE) intake via drinking water on Th17 cells in mice.
METHODSForty eight six weeks old female BALB/c mice were divided into blank control, vehicle control, 2.5 mg/ml TCE and 5.0 mg/ml TCE groups by random number table (12 mice each group), and exposed to TCE by drinking water. On the 14(th), 28(th), 56(th), 84(th) days, blood were collected and assayed for IL-17, IL-6, and TGF-β concentration in serum through ELISA. Animals were killed and spleen biopsies were taken sterility. The proportion of Th17 cells among CD4(+) T cells and RORγt mRNA expression level in spleen were measured by FCM and real-time PCR.
RESULTSIn 2.5 mg/ml TCE and 5.0 mg/ml TCE group mice, Th17 cells/CD4(+) T cells in spleen were (3.46 ± 0.32)% and (5.45 ± 0.45)% on day 14, (3.47 ± 0.33)% and (4.10 ± 0.39)% on day 84, which were significantly higher than those for solvent control group at the same time point ((2.15 ± 0.20)%, (2.16 ± 0.35)%, respectively) (P < 0.01). RORγt mRNA expression levels were (1.870 ± 0.084) and (1.965 ± 0.060) on 14 day, (1.998 ± 0.079) and (2.028 ± 0.073) on day 56, which were also significantly higher than those for solvent control group at the same time point (1.77 ± 0.04 and 1.75 ± 0.09, respectively) (P < 0.05). IL-17 concentrations in serum were (32.28 ± 5.38) and (34.47 ± 5.02) pg/ml on day 14, and (34.87 ± 5.48) and (41.94 ± 6.19) pg/ml on day 28, which were significantly higher than those for solvent control group at the same time point((21.57 ± 5.23), (22.11 ± 5.11) pg/ml). IL-6 concentration in serum were (43.07 ± 6.71) and (47.86 ± 8.52) pg/ml on day14, (41.32 ± 7.04) and (46.74 ± 9.33) pg/ml on day 56, which were significantly higher than solvent control group at the same time point ((7.56 ± 7.71) and (28.26 ± 7.22) pg/ml). TGF-β concentration were (17.48 ± 3.06) and (18.93 ± 3.12) pg/ml on day 14, which did not show significant difference from solvent control group ((15.25 ± 2.95) pg/ml). Correlation analysis showed that IL-6 in serum were significantly positively correlated with the proportion of Th17 cells among CD4(+) T cells and RORγt expression level in spleen (r = 0.741, 0.765, P < 0.01).
CONCLUSIONTCE might promote the differentiation of Th17 cells and increase IL-17 secretion by inducing IL-6 and up-regulating RORγt expression together with TGF-β.
Animals ; Drinking Water ; chemistry ; Female ; Interleukin-17 ; immunology ; Interleukin-6 ; immunology ; Mice ; Mice, Inbred BALB C ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; immunology ; Th17 Cells ; drug effects ; immunology ; Transforming Growth Factor beta ; immunology ; Trichloroethylene ; toxicity
8.Research on the distributed PACS based on grid technology.
Yong-xing LUO ; Li-jun JIANG ; Hai-bo HU ; Tian-ge ZHUANG
Chinese Journal of Medical Instrumentation 2006;30(4):287-290
In this paper, we introduce the grid technology into the design of the distributed PACS. First, we analyse the architecture and functions of OGSA-DAI, and then, introduce a model of the distributed PACS based on OGSA-DAI, and give a detailed analysis on its basic components and workflow. Finally, we make a conclusion on the distributed PACS based on grid technology.
Computer Communication Networks
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Computer Systems
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Database Management Systems
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Humans
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Information Storage and Retrieval
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methods
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Medical Informatics
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Radiology Information Systems
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Software
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Software Design
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Systems Integration
9.Helicobacter pylori infection enhances atherosclerosis in high-cholesterol diet fed C57BL/6 mice.
Xing-hai CHEN ; Jiang-bin WANG ; Yu-shan WANG ; Zhong-min LIU ; Yan LI
Chinese Journal of Cardiology 2010;38(3):259-263
OBJECTIVETo evaluate the impacts of Helicobacter pylori (H. pylori) infection on atherosclerosis and plasma lipid levels in high-cholesterol diet fed C57BL/6 mice.
METHODFemale C57BL/6 mice were randomly divided into 4 groups (n = 12 each): fed with normal chow diet (A), infected with H. pylori (B), fed with high-cholesterol diet (C) and infected with H. pylori and fed with high-cholesterol diet (D). After 52 weeks, plasma levels of lipids were measured and aortic atherosclerosis was observed. The ureA, ureC, cagA and vacA DNA were also detected by PCR in the aortic arteries.
RESULT(1) Prevalence of atherosclerosis was similar between group C and D (91.6% vs. 100%, P > 0.05) while there was no atherosclerosis in group A and B. H. pylori infected mice showed more obvious inflammation in gastric mucosa than mice without H. pylori infection. (2) The plasma levels of triglyceride, total cholesterol and LDL were higher and HDL was lower in group B, C and D than those in group A and in group D than in group C (all P < 0.05). (3) Roberts & Thompson scores and number of foam cells in plaques were significantly higher in group D compared with those in group C (all P < 0.05). (4) ureC DNA was detected in 5 out of 12 aortic arteries of mice in group D but not in group A, B and C.
CONCLUSIONOur results suggested that H. pylori infection might enhance the atherosclerotic lesion formation in this mouse model.
Animals ; Atherosclerosis ; microbiology ; pathology ; Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; Cholesterol ; blood ; Cholesterol, Dietary ; adverse effects ; DNA, Bacterial ; analysis ; Female ; Helicobacter Infections ; pathology ; Helicobacter pylori ; genetics ; Mice ; Mice, Inbred C57BL ; Triglycerides ; blood ; Urease ; genetics
10.Polymorphism of killer cell immunoglobulin-like receptors gene family in Zhejiang Han population.
Fa-Ming ZHU ; Kan JIANG ; Qin-Feng LÜ ; Wei ZHANG ; Hai-Qin ZHANG ; Qi-Hua FU ; Li-Xing YAN
Journal of Experimental Hematology 2005;13(6):1109-1112
To analyze killer immunoglobulin (Ig)-like receptor (KIR) gene content and allelic polymorphism in Zhejiang Han population, samples were genotyped by polymerase chain reaction sequence-specific primers (PCR-SSP). The results demonstrated that all 17 KIR genes could be observed in the population. All individuals contained 2DL4, 3DL2 and 3DL3 genes. The frequencies of these genes was 1.00. 2DL1, 2DL3, 2DP1, 3DP1*003, 3DL1, 2DS4*001/002 loci were more common, their frequencies were 0.902, 0.902, 0.902, 0.902, 0.7598, 0.5615 respectively, while the frequencies of 2DL2, 2DL5A, 2DL5B, 2DS1, 2DS2, 2DS3, 2DS4*003, 2DS5, 3DS1 and 3DP1*001/002 were relatively lower. The A KIR haplotype was the most prevalent (74.7%) in Zhejiang Han population and there were 12 different KIR haplotypes in total, the most common was 2 (53.0%). Twenty six different genotypes have been found in the population, AJ (2, 2) and AF (1, 2) showed higher frequencies, followed by AH (2, 5), NN2 (2, 6), AI (1, 5) and AG (1, 1). Fifteen of these genotypes have not been found in Caucasians so far and four new KIR profiles could not be assigned to the haplotypes according to standard assign method. In conclusion, there are distinctive frequencies of KIR gene content, haplotype as well as genotype in Zhejiang Han population.
China
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Gene Frequency
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Genotype
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Haplotypes
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Humans
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Polymerase Chain Reaction
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methods
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Polymorphism, Genetic
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Receptors, KIR
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genetics