Aim To study the effect of genistein on contractility of isolated papillary muscles in guinea pig and its mechanisms.Methods Isolated guinea pig papillary muscles were suspended in organ baths containing KH solution,and the effect of genistein on contractility of myocardium was observed after an equilibration period.Results Genistein and isoprenaline hydrochloride showed positive inotropic effects on contractility of myocardium in a dose-dependent manner(1～100 ?mol?L~(-1)) Propranolol(1 ?mol?L~(-1)) and verapamil hydrochloride(0.5 ?mol?L~(-1)) attenuated the positive inotropic effect of isoprenaline hydrochloride(2 ?mol?L~(-1)),but did not change the effect of genistein(100 ?mol?L~(-1)).Furthermore,the enhancement of the contractility induced by elevation of extracellular Ca~(2+) concentration in papillary muscles did not show any change after pretreatment with genistein(1,10 ?mol?L~(-1)).Finally,the positive inotropic effect of genistein was attenuated partially by tamoxifen(1 ?mol?L~(-1)),however,was not influenced by sodium orthovanadate(1 ?mol?L~(-1)).Conclusions Genistein has the positive inotropic effect on guinea pig papillary muscles,which is not related to the activation of Ca~(2+) channel on cell membrane,? adrenoceptor and tyrosine kinase pathway,but this effect may involve in uptake and utilization of sarcoplasmic reticulum Ca~(2+).

Objective:To explore the application value of dynamic monitoring of serum soluble interleukin-2 receptor (sIL-2R) and stromal cell-derived factor 1 α (SDF-1α) in patients with acute leukemia (AL).Methods:A total of 187 patients with AL admitted to the Second Hospital of Shanxi Medical University from March 2017 to August 2018 were selected and 90 healthy subjects at the same period were selected as the healthy controls. The levels of serum sIL-2R and SDF-1α were detected at the initial diagnosis, remission phase and relapse phase, respectively. The clinical value of dynamic monitoring of sIL-2R and SDF-1α was analyzed.Results:Among 187 patients with AL, 135 patients (72.19%) had complete remission (CR) after chemotherapy, 52 patients (27.81%) had partial remission, and 43 patients (31.85%) relapsed. The level of serum sIL-2R at the AL initial diagnosis period was (533±32) U/ml, which was higher than that in the healthy controls [(247±30) U/ml], and the difference was statistically significant ( t = 71.976, P < 0.01); the level of serum SDF-1α at the AL initial diagnosis was (2 968±305) pg/ml, which was higher than that in the healthy controls [(1 358±160) pg/ml], and the difference was statistically significant ( t = 47.043, P < 0.01). The levels of serum sIL-2R [(308±30) U/ml] and SDF-1α [(1 576±184) pg/ml] in the CR phase were lower than those in the initial diagnosis of patients with AL; and the levels of sIL-2R [(599±36) U/ml] and SDF-1α [(2 894±301) pg/ml] in the relapse phase were higher than those in the CR phase of patients with AL (all P < 0.01). Conclusions:The levels of serum sIL-2R and SDF-1α in patients with AL are increased, and there are big differences in the levels of sIL-2R and SDF-1α at the initial diagnosis, remission phase, and relapse phase. Dynamic monitoring of both can provide the data support for early clinical intervention.

Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

BACKGROUNDAcute lymphoblastic leukemia (ALL) and chemotherapy can cause immune imbalance, and gaseous molecule hydrogen sulfide (H2S) can participate in the process of immune response. This study aimed to investigate the immune regulation of H2S in pediatric ALL.

METHODSChildren (n = 78) with ALL admitted during 2010-2013 were included in this study. Two blood samples were collected in period of before chemotherapy, bone marrow remission and two days after chemotherapy, respectively. Serum contents of H2S and cytokines, including interleukin-1β (IL-1β), interleukin-2 (IL-2), interferon-γ (IFN-γ), tumor necrosis factor (TNF-α), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10) and macrophage inflammatory protein-1α (MIP-1α), were detected using ELISA method. Stepwise regression was used to analyze the correlation between H2S and cytokines. Furthermore, human Jurkat cells were cultured in vitro, and nucleoprotein of Jurkat cells and peripheral blood mononuclear cells (PBMCs) were collected, contents of cystathionine γ-lyase (CSE) and certain cytokines were measured by Western blotting.

RESULTSSerum concentrations of H2S, IL-1β, IL-6, IL-10 and MIP-1a in children with ALL were increased significantly (P < 0.01), while concentrations of IL-2, TNF-α, IFN-γ and IL-4 decreased obviously (P < 0.01). In patients after chemotherapy, concentrations of H2S and IL-10 were decreased significantly (P < 0.05), but IL-4 and IFN-γ concentrations increased markedly (P < 0.05). At remission stage, H2S, IL-1β, IL-4, IL-6, IL-10 and MIP-1α concentrations were further decreased markedly (P < 0.05), but concentrations of IL-2, TNF-α and IFN-γ increased again (P < 0.05). Protein contents of CSE, IL-10, IL-4 and IL-2 of PBMCs also increased markedly in children with ALL. Moreover, changes of CSE protein contents of PBMCs were consistent with serum H2S contents, and there were significant correlation between H2S and certain cytokines based on stepwise regression analysis. Furthermore, compared with those of PBMCs group, in vitro study indicated that Jurkat cells of H2S group expressed IFN-γ, IL-10, IL-4 and IL-2 protein increased obviously (P < 0.05), while IL-4, IL-2 and CSE expression of PPG group decreased markedly (P < 0.05).

CONCLUSIONGaseous molecule H2S might participate in the process of immune regulation in pediatric ALL through modulating transcription and expression of cytokines.

Child ; Child, Preschool ; Cystathionine gamma-Lyase ; blood ; Female ; Humans ; Hydrogen Sulfide ; blood ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-1beta ; blood ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; Tumor Necrosis Factor-alpha ; blood