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Objective: Glucocorticoids (GCs) are the effective first-line drugs and indispensable in chemotherapy regimens to treat patients with multiple myeloma (MM). Previous studies in a variety of hematologic malignancies have shown that the biological action of GC is mediated through the expression and activation and of glucocorticoids receptor (GR) isoforms in vitro . GR and its regulation are crucial determinants of the efficacy of GC independent therapy. There is currently lack of research on patients with MM., Methods: 132 patients with MM were divided into responders (78 cases) and nonresponders (54 cases) according to the efficacy evaluated after four cycles of GC-dependent regimen. 66 patients with iron-deficiency anemia were served as controls. Preparation of mononuclear bone marrow cells (MBMCs) was purified by Ficoll-Hypaque gradient centrifugation. The mRNA expression of GR α, β, γ, P, SRp30, SRp40, HSP90, NF-κB and AP-1 were detected by real time RT-PCR., Trial Registration: CHiCTR-RCH-12002872., Results: The expression of four GR isoforms exhibited the following trend in MM patients and controls: GRα>GR-P>GRγ>GRβ. GRα and HSP90 expression in responders was significantly higher than that of the nonresponders ( P <0.050). HSP90/GRα expression in MM patients exhibited significantly higher than that in controls ( P <0.001). SRp30c and SRp40 mRNA expression both showed significant positive correlation with GRα transcript ( P <0.001). Compared with controls, NF-kB and AP -1 expression in MM patients was higher. NF-kB and AP-1 expression of nonresponders were significantly higher than that of responders. The difference was not obvious statistically ( P >0.050)., Conclusion: Our findings raise the possibility that low expression of GRα and HSP90 plays important roles in nonresponders. Lack of HSP90 might affect GR structure and further take part in nonresponse. SRp30c and SRp40 mRNA expression both showed significant positive correlation with GRα. That might become new targets for treatment of nonresponders in MM patients, although further studies are needed for clarification., (© 2021 by the Association of Clinical Scientists, Inc.)

Background/aims: MiR-19b has been reported to be involved in several malignancies, but its role in multiple myeloma (MM) is still unknown. The objective of this study was to explore the biological mechanism of miR-19b in the progression of MM., Methods: First, we performed real-time polymerase chain reaction (PCR) and Western blot to study the expression of miR-19b, tuberous sclerosis 1 (TSC1), and caspase-3 in different groups. MTT assay was performed to explore the effect of miR-19b on survival and apoptosis of cancer stem cells (CSCs). Computation analysis and luciferase assay were utilized to confirm the interaction between miR-19b and TSC1., Results: A total of 38 participants comprising 20 subjects with MM and 18 healthy subjects as normal controls were enrolled in our study. Real-time PCR showed dramatic upregulation of miR-19b, but TSC1 was evidently suppressed in the MM group. MiR-19b overexpression substantially promoted clonogenicity and cell viability, and further inhibited apoptosis of CSCs in vitro. Furthermore, miR-19b overexpression downregulated the expression of caspase-3, which induced apoptosis. Using in silico analysis, we identified that TSC1 might be a direct downstream target of miR-19b, and this was further confirmed by luciferase assay showing that miR-19b apparently reduced the luciferase activity of wild-type TSC1 3´-UTR, but not that of mutant TSC1 3´-UTR. There was also evident decrease in TSC1 mRNA and protein in CSCs following introduction of miR-19b. Interestingly, reintroduction of TSC1 abolished the miR-19b-induced proliferation promotion and apoptosis inhibition in CSCs., Conclusion: These findings collectively suggest that miR-19b promotes cell survival and suppresses apoptosis of MM CSCs via targeting TSC1 directly, indicating that miR-19b may serve as a potential and novel therapeutic target of MM based on miRNA expression., (© 2018 The Author(s). Published by S. Karger AG, Basel.)

In this study, we first initiated a multicenter, single-arm, phase-II clinical trial using decitabine (DAC) (20mg/m 2 for five days) based chemotherapy, followed by haploidentical lymphocyte infusion (HLI) that was applied as induction therapy for elderly patients with AML. Furthermore, the role of HLI infusion was explored in a mouse model. The clinical trial included 29 elderly patients (median age: 64, range 57-77) with AML. Sixteen cases achieved complete remission (CR) and 9 cases achieved partial remission (PR) after the first treatment cycle. Of the patients with PR, 5 subjects achieved remission after the second induction, which brings the overall CR rate to 72.4%. The 2-year overall survival (OS) and disease-free survival (DFS) was 59.6% and 36.9% respectively. The treatment regimen was well tolerated with only one patient died of severe pneumonia one month after the first treatment. In the mouse experiment, we found that DAC/HLI significantly enhanced the survival of leukemic mice. These results suggested that DAC-based chemotherapy combined with HLI is an alternative first line induction therapy for elderly patients with AML. This trial is registered at ClinicalTrials.gov (NCT01690507)., Competing Interests: CONFLICTS OF INTEREST The authors declare that there is no conflict of interest.

Previous studies have indicated that leptin and the soluble leptin receptor (SLR) might influence inflammatory and immune processes in autoimmune diseases, but this remains unclear in systemic lupus erythematosus (SLE). The aim of our study was to assess if leptin and SLR are involved in the etiopathology of SLE and the possible mechanism of immune regulation. We studied 87 patients with SLE and 85 matched subjects. We assessed the levels of serum leptin and SLR, tested the long isoform leptin receptor (Ob-Rb) mRNA levels in SLE patients and a control group. Furthermore, we measured Th1 and Th2 percentage in SLE patients' lymphocytes and examined lymphocytes activation and proliferation assays with leptin stimulation in vitro. The study found a higher level of serum leptin in SLE patients, however, no difference was found in serum SLR levels or Ob-Rb mRNA levels between SLE patients and the control group. The percentage of Th1 cells decreased and Th2 cells increased after treatment with glucocorticoids in SLE patients. Leptin stimulated the proliferation of T cells in vitro, and differentiation to Th1 cells increased. The present study demonstrated that leptin may play an important role in the pathogenesis of SLE, inducing dysfunction of autoimmune processes.

Alternative splicing of the glucocorticoid receptor (GR) gene results in several GR isoforms, we examined their expression (GRα, GRβ, GRγ and GR-P) by real-time RT-PCR in glucocorticoid (GC) sensitive (CEM-C7), GC resistant (CEM-C1) cells and adult acute lymphoblastic leukemia (ALL) patients, to determine the association of GR isoform expression profiles and GC resistance in adult ALL patients. With GC treatment, GR levels in C1 cells showed no obvious changes. In C7 cells, the mRNA levels of GRα, GRβ and GRγ first increased and then decreased, whereas GR-P mRNA had a continued rising trend. C7 cells had a higher GRα/GRγ, lower GRα/GR-P and GRγ/GR-P ratios than C1 cells (P < 0.01). In adult ALL patients, GRγ mRNA varied in different ALL stages (complete remission CR 15.82 vs. relapsed 8.21 vs. initial 1.93 P < 0.05). It also did in the ratios between GR isoforms that GRα/GRγ and GRα/GR-P in initial patients were higher than relapsed and CR (P < 0.05), while GRγ/GR-P in CR was higher than initial and relapsed patients (P < 0.05). GR-P mRNA in T-ALL patients was much higher than that in B-ALL patients (P < 0.05). Peripheral blood hemoglobin (HB) was positively correlated with GRα mRNA and GR-P mRNA (P < 0.05), while white blood cells (WBC) negatively correlated with GRγ mRNA (P < 0.05). The present study demonstrates that GR autoinduction is more important to GC sensitivity than its basal level expression. GC sensitivity is also significantly correlated with GRα mRNA and mildly associated with GRβ mRNA expression. Both GRγ mRNA and the ratios between GR isoforms (GRα/GRγ, GRα/GR-P and GRγ/GR-P) are correlated with ALL stages. The changes of mRNA expression levels of GRα, GR-P and GRγ may provide valuable information for GC resistance. Peripheral blood HB and WBC affect GR isoform expression.

Previous studies have indicated that autoimmune diseases might be caused by an imbalance of T helper cells (Th), cytokines, and regulatory T cells (Treg) cytokines. We measured the plasma concentrations of Th1-associated cytokines (IFN-gamma, IL-2), Th2 -associated cytokines (IL-4, IL-10), Th17-associated cytokine (IL-17) and Treg -associated cytokine (TGF-beta1) in adult patients with immune thrombocytopenia (ITP) and evaluated their clinical relevance. Plasma IFN-gamma, IL-2, IL-4, IL-10, IL-17 and TGF-beta1 concentrations of 52 ITP patients and 30 age- and sex-matched healthy controls were measured by enzyme-linked immunosorbent assay method (ELISA). Concentration of Th2 cytokines (IL-4 and IL-10) were significantly higher in ITP patients compared to controls (P < 0.05). However, concentrations of Th1 cytokines (IFN-gamma, IL-2), Th17 cytokine (IL-17) and Treg cytokine (TGF-beta1) were lower in ITP patients (P < 0.05). Concentration of IL-17 was significantly higher in chronic ITP patients compared to severe ITP patients (P < 0.05), and no significant difference of cytokine concentration among the other subgroups in ITP patients was found. Among the ITP patients, concentration of IFN-gamma correlated positively and significantly with PAIgG (r = 0.48, P = 0.02). A significant correlation was neither found between other cytokine levels and platelet count, nor between cytokine levels and megakaryocytes number, nor between cytokines levels and PAIgG or GPIIb/IIIa and/or GPIb/IX autoantibodies. The present study demonstrates that an imbalance of Th and Treg cytokines may mediate the pathogenesis of ITP.

Glucocorticoids (GCs) are considered the important drugs used in treatment of idiopathic thrombocytopenic purpura (ITP). However, about 10-30% patients with ITP develop GC resistance after standard treatment with GC. The macrophage migration inhibitory factor (MIF) has been shown to act as a counter-regulator of the anti-inflammatory and immunosuppressive effects of GCs on immune cells. In addition, MIF-173G/C polymorphism was associated with higher MIF expression both in vitro and in vivo. In this case-control study, we investigated the association of GC resistance with MIF polymorphism and expression. MIF mRNA expression was analyzed by semiquantitative real-time RT-PCR in GC-sensitive and GC-resistant ITP patients. MIF protein expression in serum was performed by ELISA. Genotyping for the MIF -173G/C polymorphism was analyzed by a Polymerase chain reaction Tm-shift genotyping method. We found no association of GC resistance and MIF mRNA and protein expression. According to sex, age, initial blood platelet count, and disease course, the patients were further subdivided into 8 groups, no statistical difference was found. In addition, we compared the distribution of the MIF -173G/C genotype and allele frequencies between the GC-sensitive ITP patients and the GC-resistant and found no statistical difference. The present study suggested that MIF polymorphism and expression does not contribute to GC resistance in ITP.

The FKBP5 gene codes for the FK506-binding protein 51 (FKBP5), a co-chaperone of hsp90, which regulates glucocorticoid (GC) receptor sensitivity. The FKBP5 gene single nucleotide polymorphisms (SNP), rs1360780, has been found to modulate GC sensitivity in stress-related psychiatric disorders. The aim of the present study was to examine the effects of rs1360780 on the treatment outcome of patients suffering from idiopathic thrombocytopenic purpura (ITP) administered with GC. The polymorphism of FKBP5, rs1360780, was genotyped in 55 GC-resistant ITP patients, 157 GC-sensitive ITP patients and 110 unrelated healthy individuals using real-time PCR and cycling probe technology with DNA extracted from peripheral blood. No significant differences in FKBP5 rs1360780 genotypes (P=0.51) and alleles (P=0.89) were observed between the GC-resistant ITP patients and the healthy controls. There were no significant differences observed between the GC-sensitive ITP patients and the healthy controls (P=0.40 for genotypes and P=0.62 for T allele), as well as between the GC-sensitive ITP patients and the GC-resistant patients (P=0.67 for genotypes and for T allele). The present study demonstrates that the FKBP5 polymorphism may not affect the response of ITP patients to GC treatment.