Preliminary study of contrast-enhanced ultrasonography on the efficacy of knee synovial lesions in rheumatoid arthritis / 中华超声影像学杂志

Objective To investigate the effect of contrast-enhanced ultrasonography (CEUS) on the treatment of knee synovial lesions in rheumatoid arthritis (RA).Methods The results of routine ultrasonography (US) and CEUS were observed in 37 patients with RA.Among them 26 knees were underwent review after treatment.The results before and after treatment were compared.Results Routine US showed that the synovial thickness of patella,medial and lateral condylar and the depth of suprapatellar bursa effusion in 37 knee joints were (0.47 ± 0.26)cm,(0.31 ± 0.15)cm,(0.36 ± 0.21)cm and (0.72 ± 0.42)cm before treatment,and (0.36± 0.16)cm,(0.28 ± 0.17)cm,(0.30 ± 0.19)cm and (0.41 ± 0.19)cm in 26 knee joints after treatment,respectively,the differences were statistically significant(P ＜0.05).The synovial blood flow classification of patella,medial and the lateral condyle in the 26 knee joints had difference between before and after treatment (P ＜0.05).CEUS showed that the peak intensity decreased,the area under the curve reduced,the time from peak to one half decreased,the wash in slope decreased and the time to peak prolonged in synovial after treatment,the differences of the parameters between before and after treatment were statistically significant(P ＜0.05).The area of synovial had some influence on the CEUS parameters and could improve the reliability of the evaluation to CEUS for treatment.Conclusions CEUS is an objective method to evaluate the efficacy of RA,which provides a reliable basis for clinical treatment of RA.

Effects of stromal cells on sentitivity to imatinib in Sup-B15 Philadelphia chromosome positive acute lymphoblastic leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the sensitivity of imatinib (IM) on Sup-B15 Ph+ acute lmphoblastic leukemia (ALL) cells indused by stromal cells OP9, and to further explore its mechanism.</p><p><b>METHODS</b>The study is divided into two group, Sup-B15 cells group and co-cultured with OP9 cells group (Sup-B15/OP9 group). The inhibitory effects of IM on leukemia cells were measured by CCK-8 test, and the apoptosis by Annexin Ⅴ/7-AAD dyeing and the percentage of CD 34+CD38- leukemia cells were determined by flow cytometry. ALDH1, CD144, and β-catenin mRNA were detected by real-time RT-PCR, protein levels by Western blot. Inmunoprecipitation was used to detect the level of β-catenin connected to CD144.</p><p><b>RESULTS</b>IM presented inhibitory effects on Sup-B15 and Sup-B15/OP9 cells at multiple concentrations from 10 μmol/L to 45 μmol/L. The IC50 of IM on Sup-B15/OP and Sup-B15 cells were 35.8 μmol/L and 6.3 μmol/L, respectively (P<0.05). After 24 h of 30 μmol/L IM treatment, the percentages of apoptosis cells in Sup-B15/OP9 and Sup-B 15 cell were (14.24 ± 2.11)% and (3.45 ± 0.68)%, respectively (P<0.05). The percentage of CD34+CD38- cells in Sup-B15/OP9 group was significantly higher than that in Sup-B15 group [(3.42 ± 0.28)% vs (0.16 ± 0.15)%, P<0.05]. As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/OP9 group was remarkably upregulated (0.097 ± 0.012 vs 0.046 ± 0.010, P<0.05), and the CD133 protein level was also upregulated in Sup-B15/OP9 group. The transcription of CD144 in Sup-B15/OP9 group was remarkably upregulated compared with Sup-B15 group (0.103 ± 0.015 vs 0.010±0.003, P<0.05), as well as the CD144 protein. β-catenin mRNA transcription has no obvious changes between Sup-B15 group and Sup-B15/OP9 group (P>0.05), while the whole β-catenin protein and the cell nucleus β-catenin significantly increased, as well as the β-catenin protein combined with CD144.</p><p><b>CONCLUSION</b>Co-cultured with OP9 cells, Sup-B15 cells show less sensitivity to imatinib. The raising activity of CD144 and CD144/β-catenin signaling may work in this procession.</p>

In vitro study of BRD4 inhibitor GSK525762A against primary adult common B-cell acute lymphoblastic leukemia cells in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of bromodomain-containing protein 4 (BRD4) inhibitor GSK525762A on the proliferation and apoptosis of primary common B-cell acute lymphoblastic leukemia (common B-ALL) cells from adult patients, then to further explore the possible mechanisms.</p><p><b>METHODS</b>Purified leukemia cells from 14 common B-ALL adult patients (4 Ph⁺ and 10 Ph⁻ cases) were obtained by flow cytometry sorting, and maintained in a mimic bone marrow microenvironment culture system for short-term culture. Leukemia cells were treated with various concentrations of GSK525762A. The inhibitory effects of BRD4 inhibitor on common B-ALL leukemia cells were measured by CCK-8 assay and the apoptosis of those cells was determined by AnnexinⅤ/7-AAD staining using flow cytometry. The transcripts of c-MYC, CDK6 and Bcl-2 were detected by quantitative RT-PCR, and the expression of c-MYC, CDK6 and Bcl-2 proteins were detected via Western blot.</p><p><b>RESULTS</b>GSK525762A could inhibit the proliferation of leukemia cells from all 14 common B-ALL patients in a dose-dependent manner, the median value of IC50 was 256.25 (90.64-1 378.39)nmol/L. GSK525762A could promote cells apoptosis of B-ALL leukemia cells in a dose-dependent manner, the median apoptosis rates respectively were 45.17%(9.38%-70.91%), 66.02% (24.36%-96.34%) and 89.29% (39.29%-99.37%) after treated by 500, 1 000 and 2 500 nmol/L GSK525762A. GSK525762A has a similar effect on Ph⁺ ALL and Ph⁻ B-ALL, but the effect of proliferation inhibition and apoptosis enhancement on Ph+ B-ALL is weaker than that on Ph⁻ B-ALL. Compared with vehicle control group, the levels of c-MYC, Bcl-2 and CDK6 transcripts in leukemic cells were reduced after treatment for 24 h and 48 h by 1 000 nmol/L GSK525762A, and there are no significant differences in the downregulation of c-MYC and CDK6 mRNA between Ph⁺ and Ph⁻ B-ALL; however, the inhibitory effect on Bcl-2 transcription was weaker in Ph⁺ B-ALL cells than that in Ph⁻ B-ALL cells. Moreover, c-MYC, Bcl-2 and CDK6 protein levels decreased in GSK525762A treated group.</p><p><b>CONCLUSION</b>GSK525762A could strongly inhibit the proliferation of common B-ALL and trigger apoptosis; meanwhile it has certain effects against Ph⁺ ALL in vitro. The effect may be achieved by down-regulation of c-MYC, CDK6 and Bcl-2 expression.</p>

Effect of bromdomain protein 4 inhibitor GSK525762A on the proliferation and apoptosis of B-cell acute lymphoblastic leukemia cells and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of bromdomain protein 4 (BRD4) inhibitor GSK525762A on the proliferation, apoptosis of B-cell acute lymphoblastic leukemia cell line RS4;11 cells, and to further explore the mechanism.</p><p><b>METHODS</b>Compared with Jurkat leukemia cells, the activity of BRD4 on RS4; 11 cells were inhibited by the inhibitor GSK525762A. The inhibitory effects of BRD4 on RS4; 11 cells were measured by CCK-8 test and the apoptosis of those cells was determined by AnnexinV/7-AAD dyeing using flow cytometry. The transcripts of anti-apoptotic genes c-myc, Bcl-2, CDK6 and proapoptotic genes Bad, Bak, Bax were detected by quantitative PCR, and the expression of Bcl-2 and Bak proteins were detected via Western blot.</p><p><b>RESULTS</b>Proliferation of RS4;11 cells could be inhibited by GSK525762A in a time- and dose-dependent manner, and the inhibitory IC50 at 48 and 72 h was 6.174 and 1.996 μmol/L, respectively. Compared with DMSO in control group, the levels of c-myc, Bcl-2 and CDK6 mRNA transcripts in RS4; 11 cells were reduced in GSK525762A treated group, while the levels of Bad, Bak, Bax mRNA transcripts were enhanced,moreover, Bcl- 2 protein levels decreased and Bak protein levels increased. However, the inhibitory effect of GSK525762A on Jurkat cells proliferation was not obvious.</p><p><b>CONCLUSION</b>GSK525762A can inhibit the proliferation of RS4; 11 cells and promoted cells apoptosis. The possible mechanisms underlying this phenomenon might be achieved via downregulation of Bcl-2 protein induced apoptosis of leukemia cells.</p>