Effects of KIF23 Gene Silencing on Proliferation,Migration and Invasion of Human Hepatocellular Carcinoma HepG2 cells / 中山大学学报(医学科学版)

[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.

Screening disease-associated proteins from sera of patients with rheumatoid arthritis: a comparative proteomic study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation at the synovial membrane. Although great progress has been made recently in exploring the etiology and pathogenesis of RA, its molecular pathological mechanism remains to be further defined and it is still a great challenge in determining the diagnosis and in choosing the appropriate therapy in early patients. This study was performed to screen candidate RA-associated serum proteins by comparative proteomics to provide research clues to early diagnosis and treatment of RA.</p><p><b>METHODS</b>Sera isolated from 6 RA patients and 6 healthy volunteers were pooled respectively and high-abundance proteins were depleted by Plasma 7 Multiple Affinity Removal System. The protein expression profiles between the two groups were then compared by two-dimensional gel electrophoresis (2-DE) and the proteins over/under-expressed by more than 3-fold were identified by mass spectrometry analysis. To validate the differential expression levels of the identified proteins between the two groups, ELISA was performed in two of the identified proteins in individual sera from 32 RA patients and 32 volunteers.</p><p><b>RESULTS</b>Eight proteins which over/under-expressed in sera of RA patients were identified. Among them, chain A of transthyretin (TTR) was under-expressed, while serum amyloid A protein, apolipoprotein A (ApoA)-IV, ApoA-IV precursor, haptoglobin 2, ceruloplasmin (Cp), immunoglobulin superfamily 22 and HT016 were over-expressed. ELISA test confirmed that Cp expressed remarkably higher while TTR obviously lower in RA group compared with volunteer group.</p><p><b>CONCLUSION</b>There were 8 identified proteins differentially expressed between RA group and volunteer group, which might be candidate RA-associated proteins and might be promising diagnostic indicators or therapeutic targets for RA.</p>

Value of the peripheral blood B-cells subsets in patients with ankylosing spondylitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of B-cell remains an enigma in the pathogenesis of ankylosing spondylitis (AS). This study aimed to investigate the distributions of B-cells and subsets in peripheral blood of AS patients and observe their changes in etanercept-treated AS patents.</p><p><b>METHODS</b>We detected the proportions of CD19(+) B-cell, naive B-cell (CD19(+)CD27-), memory B-cell (CD19(+)CD27dim) and plasmablast (CD19(+)CD27high) in peripheral blood of 66 patients with AS (39 at active stage, 27 at stable stage; 35 patients with peripheral joint involvement, 31 patients with axial involvement alone), 30 patients with rheumatoid arthritis (RA) and 30 healthy volunteers using flow cytometry. And then we observed the changes of the above indexes of 39 active AS patients treated with etanercept in a randomized, double-blind, placebo-controlled trial.</p><p><b>RESULTS</b>(1) Percentages of CD19(+) B-cells in active or peripheral joint involvement AS patients increased more obviously than those in stable or axial involvement alone AS patients (both P = 0.001), and percentage of CD19(+)CD27high B-cells in AS patients with peripheral joint involvement was significantly higher than that in cases with axial involvement alone or healthy volunteers (P = 0.005 and 0.006, respectively); (2) The percentage of CD19(+) B-cells in AS patients was positively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores, Patient's Global Assessment (PGA) scores, total back pain scores and nocturnal back pain scores (r = 0.270, 0.255, 0.251 and 0.266, P = 0.029, 0.039, 0.042 and 0.031, respectively); (3) At week 6 and week 12, there were no statistical differences of the percentages of B-cells and subsets between etanercept group and placebo group of AS patients (P > 0.05); the percentage of CD19(+) B-cells in etanercept group was higher than that in healthy volunteers at week 12 (t = 3.320, P = 0.003).</p><p><b>CONCLUSIONS</b>Misbalance is present in B-cells and some subsets in peripheral blood of active AS patients with peripheral joint involved. B-cells might play an important role in the pathogenesis of AS patients. The high percentage of CD19(+) B-cells in active AS patients cannot be down-regulated after 12-week etanercept treatment.</p>

Changes of T lymphocyte subsets and expression of costimulatory molecule CD154 on T-cells in peripheral blood from patients with ankylosing spondylitis / 中华风湿病学杂志

Objective To study T lymphocyte subsets and expression of costimulatory molecule CD154 on T-cells in peripheral blood from patients with ankylosing spondylitis and their changes after treated with Enbrel. Methods Sixty-six patients with AS(39 active and 27 inactive, 35 axial and peripheral joint involvement and 31 axial involvement only), 30 patients with rheumatoid arthritis(RA), 30 healthy volunteers were analyzed. The expression of CD154 on CD3+ T cell as well as T-cells subsets were evaluated using flow cytometry respectively. The changes of the expression of costimulatory molecule CD154 in 39 active AS patients(Enbrel treatment or placebo treatment) were observed in a randomized, double-blind, placebothan that of healthy volunteers, and CD154 expression on CD3+ T cells in peripheral blood in AS patients was on CD3+ T cells in the peripheral blood of active AS or AS patients with peripheral joint involvement were significantly higher than those in inactive or axial involvement only AS patients(P＜0.05), and CD154 on CD3+ placebo, in AS patients group, there was significant reduction in CD154 expression on CD3+ T cells in Enbrel group at week 6(P＜0.05), there was no significant difference between Enbrel group and healthy volunteers at week 6(P＞0.05). Conclusion T lymphocyte subsets are significantly abnormal and CD154 is overexpressed on T-cells in peripheral blood of patients with AS. A six-week course of treatment with Enbrel in active AS can induce a down-regulation of the expression of CD154 on T cells.

Expression and analysis of HLA-A, B and DRB1 genes in patients with chronic myelogenous leukemia in Guangdong area / 中国实验血液学杂志

To study the gene polymorphism of HLA-A, B, DRB1 alleles in patients with chronic myelogenous leukemia and to explore the correlation of HLA with chronic myelogenous leukemia, the polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) was used to analyze the polymorphism of HLA-A, B, DRB1 alleles of 293 CML Patients and 406 randomized and synchronous blood donors (healthy and unrelated with patients) from Guangdong Han population. The results indicate that the gene frequency of HLA-A*24 in CML group was 15.53% lower than that of control group (22.09%, RR = 0.63, p = 0.005); the gene frequency of HLA-B*13 in CML group was 10.41% higher than that of control group (6.74%, RR = 1.68, p = 0.016). The gene frequency of HLA- DRB1*14 in CML group was 7.51% lower than that of control group (11.89%, RR = 0.58, p = 0.008). The differences were all statistically significant. It is concluded that the gene frequency of HLA-A*24, HLA- DRB1*14 in CML patients is significantly lower than normal people in Guangdong. The gene frequency of HLA-B*13 in CML patients is significantly higher than normal people in Guangdong. Further study is needed to make sure whether HLA-A*24 and HLA- DRB1*14 are protective gene markers for CML acquisition on Guangdong Chinese Han population and whether HLA-B*13 is a gene marker for CML susceptibility on this population.

Apoptotic effect of oridonin on NB4 cells and its mechanism / 中草药

Objective To investigate the mechanisms of oridonin inducing apoptosis on acute leukeamia NB4 cells and its mechanism. Methods NB4 cells in culture medium in vitro were given with different concentrations (8, 16, 24, and 32 μmol/L) of oridonin. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 expression was detected by Western blotting, and caspase-3 activity was assayed with colorimetric assay kit before and after apoptosis occurred. Results Oridonin (over 16 μmol/L) could inhibit the growth of NB4 cells and cause apoptosis significantly, the suppression was both in a timeand dose-dependent manner. Marked changes of apoptosis including condensation of chromatin and nuclear fragmentation were observed very clearly by Hoechst 33258 fluorescence staining and a characteristic "ladder" of DNA fragments was elicited by agarose gel electrophoresis; Western blot analysis revealed that caspase-3 was activated by the loss of caspase-3 proenzyme (32 kDa) and the appearance of its 20 kDa subunit, and that along with the apoptotic process caspase-3 activity was increased concurrently. Conclusion Oridonin can induce apoptosis in NB4 cells via activation of caspase-3. These results will provide laboratory evidence for the clinical treatment of acute leukemia with oridonin.

Study of the expression and value of CXCRI and CXCR2 in ankylosing spondylitis / 中华风湿病学杂志

Objective To study the expression and value of CXCR1 and CXCR2 on neutrophils, CD14~+ monocytes and CD3~+ T lymphocytes of peripberol blood of ankylosing spondylitis(AS)patients and to investigate the correlation between CXCR1,CXCR2 and disease activity.Methods A case control study was designed and enrolled 30 active AS,30 active rheumatoid arthritis(RA)and 30 healthy controls.The levels of CXCR1 and CXCR2 expression on neutrophils,CD3~+ T cells and CD14~+ monocytes of peripheral blood of the patients and healthy controls enrolled were measured and analyzed by flow cytometry by measuring the mean fluorescence intensity(MFI)channel.The correlations between the level of CXCR1 and CXCR2 anti disease activity or functional index of AS such as BASDAI,BASF1,ESR and CRP were analyzed.Results The MFI of CXCR1 expression on CD3~+ T lymphocytes of peripheral blood was significantly higher in AS patients (41?24)than that in RA patients(18?10)and healthy controls(19?7)(P

WT1 peptide-loaded DC triggers cytotoxic T lymphocytes and killing effects on K562 cells in vitro / 中国病理生理杂志

AIM: To study the effects of WT1 peptide-loaded dendritic cells (DC) stimulating the cytotoxic T lymphocytes (CTL) on K562 cells in vitro. METHODS: DC were generated from normal human peripheral blood mononuclear cells (PBMC) in the presence of granulocyte-macrophage colony stimulating factor(GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-?) , DC were cultured with WT1 peptides , and then triggered T cells into specific CTL. RESULTS: Most suspended cells exhibited distinctive morphological features of DCs and they stimulated proliferation of allogenic lymphocytes. Under the effector : target ratio of ~20∶1 , CTLs derived from cultures with DC and WT1 peptides were showed 86.1%?26.8% cytotoxicity against K562 cells, cytotoxicity by CTLs derived from cultures with unloaded DC against K562 cells were 47.1%?20.8% and cytotoxicity by lymphocytes were 27.7%?15.3%. Cytotoxicity by CTLs derived from culture with WT1 peptides-loaded DC were the strongest among three groups (P