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Objective:To investigate the change of apoptosis-associated genes in human lung adenocarcinoma cell lines A549/DDP cells,which were induced by the recombinant human interleukin-24(rhIL-24) combined with Cisplatin (DDP). Methods: Six genes expression level by GeXP genetic analysis system at the same time,after rhIL-24,DDP and rhIL-24+DDP were used to intervene in A549/DDP cells. Results:rhIL-24 could induce Bax gene,Caspase3 gene and Rb gene transcription up regulation;Bcl-2 gene and survivin gene transcription down regulation. But no regular change in the genes expression level of P53. Bax,Survivin and Rb were more obviously changed after rhIL-24 combined with DDP. Conclusion: RhIL-24 can induce apoptosis of A549/DDP cells through upregulated the genes expression level of Bax,Caspase3,Rb and down regulated of Bcl-2,Survivin.
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Objective:Adjuvant arthritis(AA)rats interfered with recombinant hIL-10,methotrexate(MTX)separately,we detected the changes of T cell subsets in rat′s blood and IL-17 A in rat′s serum.Methods:AA rats interfered with recombinant hIL-10, MTX and IL-10 plus MTX.The changes of the CD4+and CD8+T cell subsets were detected by flow cytometry;the Levels of IL-17A in serum of rats were measured by ELISA method.Results:Compare to normal control group ,the CD4+T cell total quantity and CD4+/CD8+T cell Proportion was significantly decreased , there were significant differences between the treatment group and the untreated group(P<0.05 ), the IL-17A of treatment group was significantly decreased , there was a significant differences ( P<0.05 ).The combination group had significant difference compared with IL-10 group.Conclusion:Recombinant hIL-10 can down-regulate the blood of AA rats in the number of CD4 +T cells and CD4+/CD8+ratio,reduce serum levels of IL-17A.The combination group compared with the IL-10 group effect on serum IL-17A more significantly,the results displayed recombinant hIL-10 plus MTX in the treatment of rheumatoid arthritis can better play the role.
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Objective:To elucidate the characterization of CD8+T cell in H1N1 influenza vaccine for children.Methods:PBMCs were isolated from 31 children aged from 3 to 6 years old who had accepted H 1N1 influenza vaccine during December 2009 to January 2010.The lymphocytes were joined with the H 1N1 influenza vaccine as experimental group and cultured .The experiment set without vaccine group as control group .At last we detected the surface molecules by FCM .The CCK-8 assay was added to detecting cellular proliferation and cellular proliferation index were detected by CCK-8.Results: CD8+T cells of PBMC in the two groups were 13.41%and 9.41%,P>0.05.CD8+CD45RAA+naive T cells in the two groups were up to more than 80%,P>0.05.The proportion of CD8+CD45ROA+memory T cells in two groups were up to 17%-19%,P>0.05.Two subsets of CD8+CD45ROA+memory T cells :CCR7+and CD62L+single positive memory T cell subsets in the experimental group were significantly lower than that of the control group,P<0.05.The CCK-8 assay was added to detect cellular proliferation .Only 51.16% of which cellular proliferation index was greater than 0.8,with none was greater than 1 in this study.Conclusion:This study showed that the CD4+T cells were low-level,naive T cells (CD8+CD45RAA+)were higher,with antigen stimulation and response.H1N1 vaccination specific memory T cells were few in number , specific memory T cell subsets were diversity , control memory cells were the main phenotypic characteristics .Cellular proliferation index showed that the proliferation of specific CD 8+T cells vaccine was poor .
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Objective To explore the association between HLA-A*31, B*40, B*58 and DRB1*16 allele polymorphisms and onset of hemorrhagic fever with renal syndrome (HFRS) in Zunyi Han population. Methods Using group study, HLA-A*31, B*40, B*58 and DRB1*16 genotyping was conducted in 100 HFRS cases and 100 controls among Han population in Zunyi area with polymerase chain reactionsequence specific primer(PCR-SSP), gene frequency (GF) and relative risk (RR) were calculated and compared. Results The results showed that the frequencies of A*3101, B*5801 and DRB1*1602 were increased in patients as compared to the healthy controls ( RR = 13. 825, x2 = 4. 296, P = 0. 038; RR =2.614,x2 =6. 133,P=0.013;RR =8.523,x2 =8. 865,P=0. 003). The frequency of B*4001 in patients with HFRS were significantly lower than that in the healthy controls( RR =0.414,x2 =6.640,P =0. 010).Conclusion These results suggest that HLA-A*3101, B*5801 and DRB1*1602 haplotypes were strongly associated with susceptibility to HFRS disease in Zunyi Han population and allele HLA-B*4001 might be associated with protection against hantaviruses infection.
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Objective To explore the correlation between HLA-A, B alleles polymorphism and hemorrhagic fever with renal syndrome (HFRS) among Han nationality in Zunyi area. Methods Using group study, HLA-A, B genotypes were conducted in 100 HFRS cases and 100 controls among Han nationality in Zunyi area with polymerase chain reaction-sequence specific primer (PCR-SSP), gene frequency (GF) and relative risk (RR) were compared and calculated. Results The frequencies of HLA-A * 31 and HLA-B * 58 alleles in HFRS cases (GF=4%,12.5%) were strikingly higher than that in the healthy controls (X2=6.380, 7.792, P<0.05;RR=18.47,2.91). The frequencies of HLA-B * 40 alleles in HFRS cases (GF=11%) were strikingly higher than that in the healthy controls (X2=6.095,P<0.01, RR=O.47). Conclusion HLA-A * 31, B * 58 genes are positively related to HFRS of Han nationality in Zunyi area, HLA-B * 40 gene is negatively related to HFRS of Han nationality in Zunyi area.
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Objective To prepare a new kind of biological agent initially,which will be used for emergent prevention or adjuvant therapy for paratyphia.Methods Paratyphia specific factor(PA-STF) was prepared in vivo and scanned with multi-wavelength using ultraviolet spectrophotometer.Then we determined the content of polypeptide and ribose with orcinol assay and modified Lowry assay respectively,followed by sterility test,pyrogen test and safety test.The immunological activity was assayed by immune protection test.Results The physico-chemical properties of PA-STF accorded with the criteria of Chinese Bioproduct Rules(2000 edition).In the immune protection test,the survival rate of mice was higher in the two experiment groups than in control group and NS group(P
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Objective To observe the changes of peripheral blood dendritic cell(DC) subgroup in amount in pulmonary tuberculosis patients.Methods DC1 and DC2 subgroups were detected in peripheral blood of 70 pulmonary tuberculosis patients by tricolor analytic method of flow cytometry.Results In active stage of pulmonary tuberculosis,DC1/PBMC,DC2/PBMC and the absolute quantity of DC1,DC2 in peripheral blood were significantly lower than healthy subjects(P0.05);The absolute quantity of DC2 in active stage of pulmonary tuberculosis was obviously lower than that in inactive stage(P
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Objective:To investigate the effect of IGN4 gene trasfection on SGC-7901 cells,and the possible mechanism for its anti-tumor effect .Methods:Ad-ING4 was obtained by gene recombination and packaging technique in vitro.The expression of ING4 mRNA and protein were analyzed by RT-PCR and western-blot respectively.The effect of Ad-ING4 on growth of SGC-7901 cells was detected by MTT.Apoptotic cells were detected by Laser Scan Co-focal Microscope (LSCM).The expression of p21 in SGC-7901 cells was analyzed by western-blot.Results:After infection with Ad-ING4,the expression of ING4 mRNA and protein were showed in SGC-7901 cells detected.The proliferation of SGC-7901 cells was inhibited significantly(P
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Objective:To establish the technique for real-time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-PCR)by DNA melting curve analysis for detecting the CDR3 shewing of TCR alpha gene repertoire in human peripheral blood.Methods:Total RNA of peripheral blood mononuclear cell(PBMC)from 4 healthy donors and 2 patients with lymphomatous leukemia were transcripted reversely into cDNA.The cDNA of 32 TRAV gene family CDR3 was amplified by FQ-PCR.Analysis of the monoclonal/oligoclonal/polyclonal CDR3 spectratyping with DNA melting curve.Results:The FQ-PCR products of 32 TRAV family CDR3 were showedas a blur land at the predicted of products size in healthy donors and parts of TRAV family CDR3 products disappeared in patients on 1.5% agarose gel by Gold-View staining.The 32 TRAV family CDR3 were showed with different frequencies by relative fluorescence quantitative in healthy donors and the patients.The CDR3 spetratyping for 32 TRAV families was showed as polyclonal peak(Gaussian distribution)in healthy donors but showed as different monoclonal/oligoclonal/polyclonal peak in the patients with lymphomatous leukemia with DNA melting curve analysis(we called "melting curve spectratyping of CDR3")Conclusion:The study suggests that the technique of "FQ-PCR with DNA melting curve analysis be convenience and celerity for detecting the CDR3 skewing of TCR alpha gene repertoire in human peripheral blood.
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Objective:To explore the association between HLA-DR,DQ allele polymorphisms and onset of epidemic hemorrhagic fever(EHF)among Han Nationality in Zunyi area.Methods:Using group study,HLA-DR and DQ genotyping was conducted in 100 EHF cases and 100 controls among Han Nationality in Zunyi area with polymerase chain reaction-sequence specific primer(PCR-SSP),GF(gene frequency)and RR(relative risk)were calculated and compared.Results:The frequency of HLA-DRB1 16 in patients with EHF was higher than in the control group(RR=3.58,?2=4.916,P=0.0266
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Abstract Objective:To explore the suppression of curcumin on leukemia cell line HL60/DAR. Methods: The suppression of curcumin onleukemia cell line HL60/DAR was studied by MTT;the apoptosis was studied by flow cytometric Annexin V/PI dual labeling technique and fluo-rescence microscope;the change in apoptosis related gene bcl-2 was studied by flow cytometry.Results: CUrcumin suppressed the growth ofHL60/DAR obviously IC50 were 8.94,5.2l,3.82 ?g/ml for 24,48,72 h respectively. Curcumin induced Hl60/DAR cells to apoptosisdeath. CUrcumin suppressed the expression of bcl-2. Conclusion: Curcumin can suppress the growth of HL60/DAR cells.
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0.05).(3) IL-17 was positively correlated to CRP(r=0.460,P 0.05).Conclusion:(1) IL-17 in patients is much higher than that in normal people,and is positively correlated to CRP,the acknowledged marker of inflammation status,but not correlated to dialysis duration,suggesting that IL-17 plays a role in pathogenesis of immune anomaly and micro-inflammation in patients with ESRD and HD;(2) There is no correlation between IL-6 and IL-17,and the mechanism of IL-17 elevation in ESRD patients needs to be further investigated.
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Objective:To investigate frequencies and polymorphism of HLA-DRB1 and DQB1 allele in the Hans of Zunyi area.Methods:Polymerase chain reaction-sequence specific primers(PCR-SSP) were used to type HLA-DRB1 and DQB1 genes of 200 unrelated healthy Han individuals in Zunyi area.Results:13 HLA-DRB1 and 7 HLA-DQB1 alleles were obtained at low resolution level in all subjects.The allele DRB1*09,DRB1*08 and DQB1*05 were showed high distributing frequencies;The allele DRB1*10 and DQB1*04 were scarcely found with low distributing frequencies.Comparied with Northern and Southern Han people,it would seem that Han people in Zunyi are more closely related to the Southern ones.The allele B*07 was scarcely found in the Southern Han with a high distributing frequency(GF=2.0%).Conclusion:HLA-DRB1 and DQB1 of Han people in Zunyi have plenty of polymorphisms.They seem to distribute in line with the Southern Han's characteristics but have their own territory feature with a high B*07 frequency.
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Objective:To study the methods of PCR ELISA in testing the RNA of platelet activating factor receptor.Methods:Extracting the total RNA of 20 normal people and 20 psoriasis patients,the PAF R and ? actin RNA assayed by RT PCR,PAF R sense labelled by Biotin and antisense labelled by Digoxin,? actin sense labelled by Biotin and antisense labelled by fluorescein.The results of PCR were tested by ELISA and Agar gel electrophoresisa.Results:The positive number of PAF R RNA was 16 by Agar gel electrophoresis and 18 by PCR ELISA in 20 normal people;The positive number of PAF R RNA was 18 by Agar gel electrophoresisa and 20 by PCR ELISA in 20 Psoriasis patients;The positive number of ? actin was 20 in two groups with two methods.Conclusion:The sense and antisense labelled by different substance in PCR ELISA has high sensitivity and the course was simple in testing the PAF R RNA.
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Objective:To investigate the mechanisms of Mitoxantrone(MTN) induced apoptosis in human acute myloid leukemia HL 60 cells.Methods:Exposured log phase growth HL 60 cells to different Mitoxantrone concentrations for different time respectively and then test the inhibitive effect by modified tertrozalium salt(MTT) assay.Morphologic evidence for apoptosis of MTN induced on HL 60 cells was determined by transmission electron microscope.The detection of MTN induced internucleosamal DNA fragmentation by agurose gele lectrophoresis.The effects on HL 60 cells proteins(Bcl 2,Bax) implicated in apoptosis were determined by Flow cytoetric.Results:(1)The antiproliferative effects were observed following exposure micromilligram level of MTN.It shows a very substantial differences in the dose and duration dependency of the observed antiproliferative and cytotoxic effects.(2)The HL 60 cells treated by MTN are observed apoptosis characteristic morphological changes such as complete cell membrane,nuclear fragmention apoptosic bodies and found about 200 bp trapezoidal belt with DNA electrophoresis.(3)The protein test results of Bcl 2 and Bax indicate that when the time of MTN treating HL 60 cells is the same,Bcl 2 protein and the ratio of Bcl 2 protein and Bax protein(Bcl 2/Bax) descend along with the ascending concentration of MTN.Bax protein has no connection with the concentration of MTN.When the concentration of MTN treating HL 60 cells are the same.The ratio of Bcl 2/Bax descends as the time of MTN treating HL 60 cells is prolonged.Conclusion:MTN has the function of inducing apoptosis on HL 60 cells with descent of Bcl 2 and Bcl 2/Bax.The regulation pathway of Bcl 2 may be a mechanism of MTN induced apoptosis on HL 60 cells.
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Detection of EHF antibody Is usually performed by using patient'sSerum.This artical reports the urine IgG antibody assay with RFC-SpA andIFA in 111 EHF patients.The positive rate was 94.6%,being nearly thesame as serum material(95.5%).And also,the urine samples can be simplycollected.