Effects of PRDX1 gene silencing on invasion and migration of human colorectal cancer SW480 cells / 中国免疫学杂志

Objective:To investigate the effects of RNA interference(RNAi)-mediated silencing of Peroxiredoxin 1(PRDX1)gene on the invasion and migration of human colorectal cancer SW480 cells.Methods: Lentiviruses negative control vector and PRDX1 RNAi were transfected respectively into colorectal cancer SW480 cells.The transfected cells were divided into PRDX1 silencing group(si-PRDX1)and negative control group(Vector).The expressions of PRDX1 mRNA and protein in SW480 cells were exa mined by quantitative real-time PCR(qRT-PCR)and immunoblotting(Western blot),respectively.The cell migration and invasion capabilities were evaluated with transwell chamber assay and transwell chamber,respectively.The protein expressions of TIMP-2,MMP-2 and MMP-9 were detected by Western blot.Results: Compared with control group,the expressions of PRDX1 mRNA and protein were significantly decreased in PRDX1 silencing group(P<0.01),PRDX1 gene silencing cell line was successfully constructed.The levels of invasion and migration capacities of SW480 cells transfected with si-PRDX1 were lower than those in the cells transfected with control-siRNA(vector)(P<0.01).The expression of TIMP-2 was significantly increased,while the expressions of MMP-2 and MMP-9 were significantly decreased(P<0.05).Conclusion: Silencing of PRDX1 inhibits the invasion,migration and metastasis of human colorectal cancer SW480 cells by regulating the expressions of TIMP-2,MMP-2 and MMP-9.

Study of expression and regulation of TLR2/4 in mycobacterium tuberculosis heat shock proteins 16. 3 effect on mouse bone marrow-derived macrophages / 中国免疫学杂志

Objective:To study the expression and regulation of TLR2/4 in mycobacterium tuberculosis heat shock proteins 16. 3 (mycobacterium tuberculosis heat shock proteins 16. 3,MTB Hsp16. 3) effect on mouse bone marrow-derived macrophages in vitro. Methods:Bone marrow cells were isolated from tibia and femurs of BALB/c mice and incubated with GM-CSF,then detected the expression of CD11b and F4/80 with flow cytometry and observed morphology. The M0 macrophages were stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4 in intracellular at different time point. Silencing macrophages cell surface TLR2/4 molecules by siRNA technology which stimulated with MTB Hsp16. 3 for 0 h,12 h,24 h,36 h,48 h and 72 h. Real-time PCR detected the expression of TLR2/4,Ym-1,Fizz1,IL-10,TNF-α,iNOS and TGF-βin intracellular at different time point. Results:Morphology analysis showed that MTB Hsp16. 3 stimulated macrophages were round cells stretching out pseudopodia,whereas MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages had elongated fibroblastoid. Real time PCR detected the expression of TLR2/4 were upregulated after MTB Hsp16. 3 stimulated M0 macrophages. MTB Hsp16. 3 stimulated silencing TLR2/4 macrophages the expression of IL-6, TNF-α, iNOS were upregulated, whereas IL-10, TGF-β, Ym-1 and Fizz1 were downregulated. Conclusion:MTB Hsp16. 3 may stimulated M0 macrophages to M2 macrophages and suppress M1 macrophages through binding with TLR2/4 receptor,which may be involved the progresss of MTB evaded macrophage phagocytosis.

Effects of miR-581 overexpression on proliferation of human colorectal cancer SW620 cells / 中国免疫学杂志

Objective:To investigate the role of miR-581 overexpression on the proliferation of human colorectal cancer cell line SW620. Methods:The expression group,colorectal cancer SW620 cells were transfected with recombinant lentivirus vector ( LV-miR-581) and miR-581 mimics(miR-581),the negative control group were transfected with negative control lentiviral vector (LV-GFP) and negative control mimics (vector). The mRNA expression of miR-581 was identified by qRT-PCR. Proliferation of the cells were detected by CCK8 assary and colony forming assary. Results:The expression of miR-581 at mRNA significantly increased in LV-miR-581 group compared with control groups were detected by qRT-PCR ( P<0. 05 ) . Up-regulation of miR-581 markedly enhanced human colorectal cancer SW620 cells proliferation than those in the cells transfected with control vector ( P<0. 05 ) . Conclusion: Forced expression of miR-581 accelerates the proliferation of colorectal cancer SW620 cells.

Expression and significance of Gab2 in colorectal cancer tissues / 中国免疫学杂志

Objective:To investigate the expression and significance of Gab 2 in colorectal cancer tissues .Methods:Immuno-histochemistry was used to detect the expression of Gab 2 in 78 cases of colorectal cancer tissues and 46 cases of the adjacent tissues and to analyze the association of Gab2 expression with the clinicopathologic features of colorectal cancer;The expression of Gab2 in samples from 10 cases of colorectal cancer tissues and matched adjacent nontumorous tissues was detected by Western blot .Results: The results of immunohistochemistry demonstrated that the Gab 2 protein positive expression rate in 78 cases of colorectal cancer was 53.85%;whereas was negative expression or weak in the adjacent tissues , showing a significant difference of comparison within this result (P0.05) .Western blot showed that the Gab2 protein expression of colorectal cancer cases was significantly higher than that of matched adjacent nontumorous tissues ( P<0.05 ).Conclusion: Gab2 was overexpressed in colorectal cancer .Gab2 maybe play an important role in carcinogenesis and progression of colorectal carcinoma .

Effects of Gab2 overexpression on proliferation and migration of human colorectal cancer cell line SW480 / 中国免疫学杂志

Objective:To investigate the effects of Gab2 overexpression on the proliferation and migration of human colorectal cancer cell line SW480.Methods: The experimental group (LV-Gab2-GFP group),colorectal cancer SW480 cells were transfected with recombinant lentivirus vector (LV-Gab2-GFP),the negative control group was transfected with negative control lentiviral vector ( LV-GFP) ,and the blank control group without any treatment.The mRNA and protein expression of Gab 2 in cells were identified by RT-PCR and Western blot respectively.Proliferation of the cells was detected by CCK-8 colorimeter and colony forming assay.Wound-healing assay was used to determine the cells migration .Results: RT-PCR and Western blot demonstrated that Gab 2 mRNA and protein expression significantly increased in LV-Gab2-GFP group compared with control groups;overexpression of Gab2 markedly enhanced human colorectal cancer SW 480 cells proliferation and migration compared with control groups .Conclusion:Overexpression of Gab2 accelerates human colorectal cancer SW 480 cells proliferation and migration.

Comparing TCR beta chain variable gene profile skewness between children with tuberculosis and BCG-vaccinated children

We compared the T cell antigen receptor [TCR-BV] gene families of peripheral blood mononuclear cells [PMBC] between children with tuberculosis [TB] and those inoculated with the Bacille Calmette Guerin [BCG] vaccine. The total RNA was extracted from PMBC of 15 TB children, 15 BCG-vaccinated children and 15 healthy controls. The RNAs were reverse-transcribed and amplified by polymerase chain reaction [PCR]. PCR products were separated on 1.5% agarose gel and analyzed with the Genescan technique. Some TCR-BV gene families in TB children and BCG-vaccinated children exhibited a blur band in the predicted position on 1.5% agarose gel, some showed a distinct or fainted band. In general, many shared predominant clonal TCR-BV gene families [V beta2, V beta16, V beta21, V beta22] and the restricted-expression families [V beta14 and V beta17]. All the gene families of the control children only exhibited blur bands and polyclonal. The skewed profile of TCR-BV gene families in TB children and BCG-vaccinated children are similar, which may probably explain the protective effects of BCG-vaccine against TB in children

Association of HLA-A * 3101, B * 4001, B * 5801 and DRB1 * 1602 alleles with hemorrhagic fever with renal syndrome disease in Han population of Zunyi province / 中华微生物学和免疫学杂志

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.

The expression of monocyte chemoattractant protein-1 and CCR2 in tuberculous pleurisy / 中国综合临床

Objective To investigate the expression of monocyte chemoattractant protein-1(MCP-1)and CCR2 in peripheral blood and pleural fluid of tuberculous pleurisy patients.Methods Flow cytometry was used to detect the expression of CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients;ELISA was used to detect the content of MCP-1 in serum and pleural fluid.Results MCP-1 in surem and pleural fluid and CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients was significantly higher than that of the healthy normal controls[MCP-1:(340.8±220.8)and(9.0±3.8)ng/L,P＜0.01;CCR2(18.2±10.1)%and(6.9±3.5)%,P＜0.05];Both MCP-1 and CCR2 were detected in pleural fluid and both of them were positivley correlated(r=0.227,P＜0.05).Conclusion The expression of MCP-1 and CCR2 in peripheral blood of tuberculous pleurisy patient are significantly elevated which are significance molecule participating in the pathogenesis of tuberculous pleurisy.

Correlation between the HLA-A, B alleles polymorphism and hemorrhagic fever with renal syndrome of Han nationality in Zunyi area / 中华微生物学和免疫学杂志

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.

Detection and clinical significance of serum interferon-inducible protein-10 and interferon-γ in patients with severe hepatitis B / 中华传染病杂志

Objective To explore the relationship between serum levels of interferon-inducible protein-10 (IP-10), interferon-γ (IFN-γ) and hepatic inflammatory reaction, disease progression in patients with severe hepatitis B (SHB). Methods Sera of 40 patients with SHB at time of admission,at the beginning of single plasma exchange (PE), at time of PE completion and 5 days after PE. The SHB patients were divided into improved group and aggravated group. And 20 patients with chronic hepatitis B (CHB) and 20 healthy controls were enrolled in this study. Serum levels of IP-10, IFN-γand tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Results The serum levels of IP-10 in patients with SHB and CHB on admission were (683.6 174.6)ng/L and (216.1 102.9)ng/L, respectively, which were notably higher than those in healthy controls [(107.6 55.8)ng/L F=9.036, both P＜0. 01],and those in patients with SHB was significantly higher than that in patients with CHB (P＜0. 01). The serum level of IFN-γ in patients with SHB and CHB on admission were (19. 8 8. 8) ng/L and (16. 7 7. 8) ng/L,respectively, which were significantly higher than those in healthy controls [(2.6 1.2) ng/L F=9. 288, both P＜0. 01]. The serum level of IP-10 and IFN-γ were both positively correlated with TNF α (r=0. 366 and r=0. 365, respectively;P＜0.05) and both negatively correlated with prothrombinase activity (r=-0.401 and r=-0.350, respectively;P＜0.05), but not correlated with serum total bilirubin(r=0. 223 and r=0. 219, respectively;P＞0.05). The serum level of IP-10 and IFN-γ were positively correlated ( r= 0. 602 ; P= 0. 000 ). On day 5 after PE, serum level of IP-10 in patients with SHB was significantly decreased compared with that'in patients before PE (t= 8. 947, P＜0.01 in improved group;t=4. 121, P＜0.05 in aggravated group) and that in aggravated group was significantly higher than improved group (t=7.862, P＜0.01). But serum level of IFN-γ was not decreased significantly (t=0. 491, P＞0.05). Conclusions IP-10 and IFN-γ are involved in the hepatic immunopathological mechanism. Serum level of IP-10 is correlated with the severity of hepatic inflammatory injury and IP-10 could reflect the progression and development of disease in patients with SHB.

Preparation and standardization of paratyphi a specific transfer factor / 西安交通大学学报(医学版)

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

Changes of peripheral blood dendritic cell subgroup in pulmonary tuberculosis patients / 第三军医大学学报

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

An association of the HLA-DR and DQ polymorphisms with EHF in Han Nationality at Zunyi area / 中国免疫学杂志

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

Research on genetic polymorphisms of HLA-DRB1 and DQB1 in the Hans of Zunyi / 中国免疫学杂志

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.

Construction of prokaryotic expression vector of mouse CD25 extracellular domain / 第三军医大学学报

Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.