OBJECTIVE:To evaluate the cost and effectiveness of two chemotherapy regimens in the treatment of initially di-agnosed multiple myeloma (MM). METHODS:63 patients who were newly diagnosed as MM without transplant chance in our hospital were analyzed retrospectively,and then divided into MPT group (alkeran+metacortandracin+thalidomide,41 cases) and VDT group (bortezomib +dexamethasone+thalidomide,22 cases) according to chemotherapy regimens. Clinical efficacy,survival time and ADR of 2 groups were observed,and cost-effectiveness analysis of them were conducted. RESULTS:The overall re-sponse rates of MPT group and VDT group were 63.4% and 77.3%,respectively(P=0.260);the complete response rate of them were 12.2% and 36.4%,with statistical difference(P=0.024). The progression free survival of MPT group and VDT group were (14.314±0.488)months and(18.557±0.848)months,with statistical significance(P<0.001);the overall survival of MPT group and VDT group were(33.536±1.143)months and(41.048±1.868)months,with statistical significance(P=0.007). Leukocytope-nia,nausea and vomit and peripheral neurotoxicity of VDT group were both higher than those of MPT group,with statistical signif-icance (P<0.05). The cost-effectiveness ratio of MPT group was 247.48,and that of VDT group was 2 922.77;the incremental cost-effectiveness ratio was 15 125.18. The results were consistent with the sensitivity test. CONCLUSIONS:Clinical efficacy and survival time of VDT regimen is better than that of MPT regimen,but MPT regimen is superior to VDT regimen in respect of cost-effectiveness analysis and the incidence of ADR. which will more suitable for the MM patients without transplant chance or goood economic conditions.

Objective To investigate the role of IL-6/STAT3 signaling in Th17/Tr imbalance of Kawasaki disease(KD). Methods Forty-eight children with KD and eighteen age-matched healthy children were consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of IL-17A, IL-17F, RORγt, Foxp3, SOCS1 and SOCS3 were assessed by real-time PCR. The proportion of CD4+CD25+Foxp3+ regulatory T(Tr) cells and mean fluorescence intensity(MFI) for phosphorylated-STAT3(pSTAT3) protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCS1 exon2, three potential bind sites for STAT3 in 5'-untraslated region(5'-UTR) of SOCS3 in CD4+ T cells. Results (1)Compared with healthy volunteers, plasma IL-6 concentration and MFI for pSTAT3 in CD4+ T cells were elevated significantly during acute phase of KD[IL-6:(54.02±20.58) pg/ml vs (8.72±2.06) pg/ml, P<0.05;pSTAT3 MFI:(55.41±15.08) vs (9.35±3.76), P<0.05], and the two items in KD patients with coronary artery lesion (KD-CAL+) were found to be higher than those in KD patients without coronary artery lesion (KD-CAL-)[IL-6:(84.76±29.35) pg/ml vs (38.65±13.76) pg/ml, P<0.05;pSTAT3 MFI:(72.36±16.81) vs (46.93±13.57), P<0.05]. (2)Transcription levels of IL-17A, IL-17F and RORγt in patients with KD were significantly elevated (P<0.05) while the proportion of CD4+CD25+Foxp3+ Treg and expression levels of Foxp3 were detected to be lower than those in normal controls (P<0.05). The mRNA levels of IL-17A, IL-17F and RORγt in KD-CAL+ group were higher than those in KD-CAL- group(P<0.05), as well as expression level of Foxp3 were found to be lower in KD-CAL+ group(P<0.05). (3)The mRNA levels of SOCS1 and SOCS3 in CD4+ T cells increased significantly during acute phase of KD(P<0.05), while the two items in KD-CAL+ group were lower than those in KD-CAL- group(P<0.05). Furthermore, CpG islands in SOCS1 exon2 and the third potential bind site for STAT3 in SOCS3 5'-UTR were hypomethylated in acute KD, while those in healthy controls were fully demethylated(P<0.05). Demethylation levels of SOCS1 exon2 and the third potential bind site for STAT3 in SOCS3 5'-UTR in KD-CAL+ group were lower than those in KD-CAL- group(P<0.05). CpG islands in the other two bind sites for STAT3 in SOCS3 5'-UTR were fully demethylated among all the groups(P>0.05). ConclusionAberrant activation of IL-6/STAT3 signaling caused by hypomethylation of SOCS1 and SOCS3 might be one contributing factor to unbalance of Th17/Tr in KD.

Objective To investigate the effect of SOCS1 and SOCS3 hypomethylation on homeostasis of Th1/Th2 in Kawasaki disease(KD). Methods Thirty-six children with KD and sixteen age-matched healthy children consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of SOCS1, SOCS3, T-bet, IFN-γ, GATA3 and IL-4 were assessed by realtime PCR. The proportion of Th1 and Th2 cells, and mean fluorescence intensity(MFI)for phosphorylated STAT3(pSTAT3)protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCSl exon2, and three potential binding sites for STAT3 in 5'-untraslated region(5'-UTR)of SOCS3 in CD4+T cells. Comparisons between groups were performed with t-test. Results ①Compared with healthy volunteers, plasma IL-6 concentration[(51.8±16.3)pg/ml vs(8.6±2.0)pg/ml, respectively]and MFI for pSTAT3[(52±14)vs(10±4), respectively]in CD4+ T cells were elevated significantly during acute phase of KD(P＜0.05), and the two items in KD patients with coronary artery lesion(KD-CAL+)were found to be higher than those in KD patients without coronary artery lesion(KD-CAL-)[IL-6:(87.2±27.4)pg/ml vs(36.2±12.8)pg/ml, P＜0.05; pSTAT3 MFI:(75±15)vs(42±11), P＜0.05]. ② The proportions of Th1 and Th2 cells and transcription levels of Th-associating factors(T-bet, IFN-γ, GATA3 and IL-4)in CD4+ T cells increased significantly in acute KD(P＜0.05), while the rate of Thl div Th2 in KD patients was found to be lower than that in normal controls(P＜0.05). In addition, the proportions of Th1 and Th2 cells and expressions levels of Th-associating factors in KD-CAL+ group were higher than those in KD-CAL-group, as well as the rate of Thl div Th2 cells in KD -CAL+ group were lower than that in KD-CAL- group(P＜0.05). ③ The mRNA levels of SOCSl and SOCS3 in CD4+ T cells increased significantly during acute phase of KD(P＜0.05), while the two items in KDCAL+ group were lower than those in KD-CAL- group(P＜0.05). Furthermore, CpG islands in SOCSl exon2 and the third potential binding site for STAT3 in SOCS3 5'-UTR were hypomethylated in acute KD, while those in healthy volunteers were fully demethylated(P＜0.05). Demethylation levels of the two items mentioned above in the KD-CAL+ group were lower than those in the KD-CAL-group(P＜0.05). CpG islands in the other two binding sites for STAT3 in SOCS3 5'-UTR were fully demethylated among all the groups(P＞0.05).Conclusion Relative insufficiency of SOCS1 and SOCS3 expression caused by hypomethylation may be one contributing factor for the imbalance of Th1/Th2 in KD.

Objective To investigate the methylation status of Foxp3 gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Thirty children with KD and eighteen agematched healthy children consented to participate in this study. Quantitative methylation specific polymerase chain reaction(MSQP) was used to assess the methylation status of Foxp3 promoter and regulatory T cells specific demethylated region(TSDR) in CD4+ T cells. The proportion of CD4+ CD25 + Foxp3 + regulatory T (Tr) cells was analyzed by flow cytometry. Transcriptional levels of CD4+ CD25 + Foxp3 + Tr associating genes (Foxp3, CTLA4, GITR, LAG3 and CCR8 ) and Foxp3-dependent molecules (UBD and LGAIS3)were measured by real-time PCR. Results ( 1 ) Demethylation level of Foxp3 promoter in CD4 + T cells from patients with KD was lower significantly than that of health subjects( P ＜ 0.01 ), and increased significantly after treated with intravenous gamma globulin therapy(IVIG) (P ＜ 0.01 ). No difference of demethylation level of TSDR region was observed among all groups ( P ＞ 0. 05 ). ( 2 ) The proportion of CD4 + CD25 + Foxp3 + Tr in peripheral blood from patients with KD , as well as mRNA levels of Foxp3 gene in CD4+ T cells, was significantly lower than those of health subjects ( P ＜0. 01 ), and increases significantly after IVIG therapy (P ＜0.01 ). Significant positive correlations between demethylation level of Foxp3 promoter and the proportion of CD4 + CD25 + Foxp3 + Tr , or expression levels of Foxp3 in CD4 + T cells, were observed during acute phase of KD (CD4+CD25+ Foxp3+ Tr: r=0.76, P＜0. 01; Foxp3: r=0.89, P＜0. 01). (3)Transcription level of CD4+ CD25+ Foxp3+ Tr associating factors, such as CTLA4, GITR, LAG3 and CCR8, was significantly down-regulated in acute phase of KD(P＜0. 01 ), and up-regulated to some extent after treated with IVIG(P ＜0. 01 ). Expression levels of Foxp3-dependent molecules UBD and LGALS3 in CD4 + T cells decreased significantly during acute phase of KD (P ＜ 0.01 ), and basically recovered to the levels of health subjects( P ＜ 0.01 ). Conclusion Decrease of demethylation level of Fopx3 promoter is correlated with immune dysfunction in Kawasaki disease.

Objective To investigate the treatment of recombinant human interleukin-11(rhIL-11)to chemotherapy-induced thrombocytopenia in acute leukemia(AL).Methods 42 AL patients whose platelet count dropped below 20×109/L after chemotherapy received rhIL-11 by 1.5 mg daily until the platelet count was increased above 40×109/L.The efficiency of chemotherapy to 17 newly diagnosed acute myelocytic leukemia(AML)patients was evaluated after receiving two periods of chemotherapy.35 AL patients and 15 newly diagnosed AML patients were used as controls.Results The mean time of platelet count increasing from 20×109/L to above 40×109/L Was shorter in treating group (9.8±2.7)d than in control group(14.6±4.8)d .The number of patients whose platelet＜15×109/L was less in treating group than in control group after second chemotherapy,and the minimum mean count of platelet Was higher in treating group(23.5±18.3)×109/L than that in control group(10.2±9.8)×109/L .CR and CR+PR rate were not different between treating group and control group. Conclusion rhlL-11 can safely and effectively promote chemotherapy-induced platelet recovery in patients of acute leukemia with persistent affection.

Objective To investigate the effects of polycyclic aromatic hydrocarbons ( PAHs) on regulatory T ( Treg) cells in newborns.Methods Blood samples were taken from the umbilical cord of sev-en newborn babies.CD4+CD25+T cells were isolated and treated with or without 300 nmol/L of phenan-threne for 72 hours in the presence of 100 IU/ml of IL-2.The expression of forkhead box P3 (Foxp3), sig-nal transducer and activator of transcription 3 (STAT3) and STAT5 were analyzed by 8-color flow cytometry. RT-PCR was performed to detect the expression of DNA ( cytosine-5 )-methyltransferase 1 ( DNMT1 ) , DNMT3a, DNMT3b and IL-4 at mRNA level.Pyrosequencing in combination with bisulphite sequencing was used to evaluate the methylation within the promoter and the Treg-specific demethylated region ( TSDR) of Foxp3 locus.Treg cells were cultured for 7 hours with autologous Tresp at Tresp/Treg ratios of 1 ∶1, 2 ∶1, 4 ∶1 and 8 ∶1 and stimulated with anti-CD3/CD28 beads and IL-2 for the evaluation of the immunosuppres-sive activities of Treg cells.Results (1) PAHs inhibited the expression of Foxp3 and the function of Treg cells collected from newborns.(2) PAHs significantly decreased the expression of STAT5 and Foxp3, but increased the expression of STAT3 (P<0.05).(3)PAHs enhanced the methylation of the promoter and the TSDR within Foxp3 gene.(4) The transcription levels of DNMT1, DNMT3a and DNMT3b in PAHs treated group were significantly higher than those of the control group (P<0.05).(5) More IL-4 was secreted by PAHs treated CD4+CD25+T cells, indicating that IL-4 was negatively correlated with STAT5, but positively correlated with STAT3.Conclusion PAHs decreased the number and inhibited the function of Treg cells in newborns.The possible mechanism might be related to the abnormal expression of STAT3 and STAT5 in-duced by IL-4 as well as the methylation within Foxp3 gene.

Objective To investigate the pathogenesis,clinical feature,laboratory examination characteristics and the prognosis of the bone marrow necrosis.Methods To analyze the clinical data of one case of diagnostic bone marrow necrosis and review the relevant literature.Results The peripheral blood examinations were as follows:the hemoglobin(HGB)level was 36g/L,platelet count was 17 ×109 /L.The biochemistry tests showed that lactate dehydro genase (LDH)was 1 454.9U /L and alkaline phosphatase(ALP)was 1 319.4U /L.Bone marrow necrosis was detected by bone marrow smear.Bone marrow biopsy was considered as a bone marrow metastatic carcinoma,which was prone to adenocarcinoma.Conclusion The bone marrow necrosis is mainly caused by the cancer,the serious infection and the drug.Its main performances were bone pain,fever,progressive decline in bloods cells,the LDH and ALP increasing and poor prognosis.In order to extend the lifetime of this kind of patients,the key lies in the early detection,the early diagnosis,and the early treatment of its primary diseases.

Objective To investigate the changes and significance of ubiquitination of Foxp3 pro-tein during the acute phage of Kawasaki disease ( KD) .Methods Forty-eight children with KD and twenty-eight age-matched healthy children were recruited in this study.Co-immunoprecipitation and Western blot assays were performed to determine the poly-ubiquitination status of Foxp3 in CD4+T cells.The percentages of CD4+CD25high Foxp3+regulatory T cells ( Treg) and the levels of IL-10, transforming growth factor-β( TGF-β) , cytotoxic T lymphocyte-associated protein-4 ( CTLA4 ) , STIP1 homology and U-Box containing protein 1 (STUB1), heat shock protein 70 (HSP70), ubiquitin-specific-processing protease 7 (USP7) and pSTAT3 were analyzed by flow cytometry analysis.Quantitative real-time PCR was used to evaluate the tran-scription levels of TLR1-10, IL-1R1, IL-1RAP, IL-6Rα, gp130, TNFR1, MyD88 and RIP1 in CD4+T cells.Plasma concentrations of IL-1β, IL-6 and TNF-αwere measured by enzyme-linked immunosorbent as-say.Results (1) The percentages of Treg cells and the levels of IL-10, TGF-β, CTLA4 and forkhead box P3 (Foxp3) in patients with acute KD were lower than those of healthy subjects (P<0.05), while the poly-ubiquitination of Foxp3 protein was significantly enhanced in patients with acute KD [(0.52 ±0.19) vs (0.08±0.02),P<0.05].Meanwhile, the four former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those of KD patients without coronary artery lesions (KD-CAL-) (P<0.05), while the polyubiquitination level of Foxp3 protein in KD-CAL+group was much higher than that of KD-CAL-group [(0.70±0.28) vs (0.43±0.17), P<0.05].The levels of Treg cells, IL-10, TGF-βand CTLA4 in patients with KD were increased and the ubiquitination of Foxp3 protein was inhibited [(0.24±0.10) vs (0.52±0.19), P<0.05] upon the treatment with IVIG.(2) The levels of STUB1 and HSP70 in CD4+T cells were significantly elevated during acute KD, while the levels of USP7 were decreased (P<0.05).The ratios of STUB1/USP7 in patients with acute KD were much higher than those of the control group [(2.65± 0.92) vs (1.09±0.37), P<0.05], but were significantly decreased after IVIG therapy [(1.46±0.53) vs (2.65±0.92), P<0.05].A negative correlation was found between STUB1/USP7 ratio and Foxp3 level during acute KD (r=-0.56, P<0.05).Moreover, KD patients with CAL+showed higher levels of STUB1 and HSP70 and higher ratios of STUB1/USP7 (P<0.05), but lower levels of USP7 as compared with those of KD-CAL-group (P<0.05).(3) The plasma concentrations of inflammatory cytokines (IL-1β, IL-6 and TNF-α), the levels of surface receptors ( IL-1R1/IL-1RAP/TLR4, IL-6Rα/gp130 and TNFR1) and its downstream molecules ( MyD88, pSTAT3 and RIP1) in CD4+T cells were up-regulated during acute KD ( P<0.05), especially in patients with CAL+, but were down-regulated upon the IVIG therapy (P<0.05).No significant differences with other TLRs were found among the groups (P>0.05).Conclusion Hyper-ubiq-uitination of Foxp3 protein might be involved in the immune dysfunction during Kawasaki disease.

Objective To investigate the changes and significances of inducible IL-35-producing regulatory T cells(iTR35) in immunological pathogcnesis of Kawasaki disease (KD).Methods Forty-eight children with KD and 32 age-matched healthy children (healthy control group) consented to participate in this study.Flow cytometry was performed to evaluate the proportions of CD4+ FOXP3-IL-12p35+IL-27EBI3+iTR35 and CD4+CD25high FOXP3+regulatory T cells (Treg),and expression levels of associated molecules such as programmed death-ligand 1 (PD-L1),CD169,programmed death 1 (PD-1),CD43,IL-12p35,Epstein-Barr virus induced 3 (IL-27EBI3),glycoprotein 130(gp130),IL-12 receptor beta 2 (IL-12Rβ2),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 4 (pSTAT4).Transcription levels of the Sre homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),Vavl guanine nucleotide exchange factor(Vav) in CD4+T cells were determined by quantitative real-time PCR.Plasma concentrations of IL-35,IL-10,TNF-α and IL-12 were measured by enzyme-linked immunosorbent assay.Results (1) The proportions of iTR35 and its expressions of IL-12p35 and IL-27EBI3 in patients with acute KD dccreased remarkably[iTR35:(0.72±0.26) ‰ vs (1.65±0.43) ‰,P＜0.05],and restored after treatment [iTR35:(1.58±0.63) ‰ vs (0.72±0.26) ‰,P＜0.05].(2) The proportions of Treg and transcriptional levels of IL-12p35 and IL-27EBI3 were down-regulated during acute phase of KD [Treg:(3.26±1.21) ％ vs (7.26±2.86) ％,P＜0.05],and increased to some extent after therapy [Treg:(5.89±2.60)％ vs (3.26±1.21)％,P＜0.05].Meanwhile,plasma concentrations of IL-35 and IL-10,and expressions of gp130,IL-12Rβ2,pSTAT1 and pSTAT4 in iTR35 of patients with acute KD were found lower than those of the healthy control group (all P＜0.05),and increased after treatment (P＜0.05).Additionally,positive correlations were found between plasma concentrations of IL-35 and the proportion of iTR35 or its expressions of IL-12p35 and IL-27EBI3,respectively.(3) Expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated significantly during acute KD(all P＜0.05),as well as expression levels of the ligands (PD-1 and CD43) and its downstream molecules (SHP-2,PTEN,Vav) in CD4 + T cells were found to be lower in patients with acute KD (P＜0.05),and restored remarkably after therapy.Conclusion Insufficiency of iTR35 and its expression of IL-35 might be one of the important factors contributing to immunological dysfunction in KD.

Objective To investigate the role of IL-35-producing regulatory B cells(IL-35 + Breg)in immunological pathogenesis of Kawasaki disease (KD).Methods Thirty-two children with KD and 28 age-matched healthy children were allowed to participate in this study.Flow cytometry was performed to evaluate the proportions of IL-35 + Breg as well as requlatory T cells (Treg)and expression levels of associated molecules such as programmed death-ligand 1 (PD-LI),CD169,programmed death 1 (PD-1),CD43,IL-12p35,epstein-Barr virus induced 3 (IL-27 EBI3).IL-12 receptor beta 2 (IL-12 Rβ2),IL-27 receptor alpha (IL-27 Rα),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 3 (pSTAT3).Transcription levels of the Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),vav1 guanine nucleotide exchange factor(Vav) in CD19 + B cells were determined by using quantitative real-time PCR.Plasma concentrations of IL-35,tumor necrosis factor α(TNF-α) and IL-12 were measured by adopting enzyme-linked immunosorbent assay.Results (1) The proportions of IL-35 +Breg and its expressions of IL-12p35,IL-27EBI3 and IL-10 in patients with acute KD were lower than those of healthy controls [IL-35 + Breg:(5.79 ± 2.60) ％ vs (12.65 ± 5.34) ％;F =19.23,9.70,14.30.7.08;all P ＜ 0.05],but they were significantly increased after intravenous immune globulin (IVIG) treatment [IL-35 + Breg:(10.52 ± 4.95) ％;all P ＜ 0.05].(2) The proportions of Treg and its transcriptional levels of IL-12p35 and IL-27 EBI3 were down-regulated during acute KD [Treg:(4.12 ± 1.51) ％ vs (8.06 ± 3.32) ％;F =19.70,17.69,38.22;all P ＜ 0.05],but were increased after therapy [Treg:(7.39 ± 2.85) ％;P ＜ 0.05].A positive correlation was found between the proportions of Treg and IL-35 + Breg during acute KD (r =0.69,P ＜ 0.05).Meanwhile,plasma concentrations of IL-35 and expression levels of IL-12Rβ2,IL-27Rα,pSTAT1 and pSTAT3 in CD19 + B cells were significantly down-regulated in children with acute KD,but they were increased after treatment(F =8.09,7.54,7.69,5.89,12.59,all P ＜ 0.05).(3) Compared with healthy controls,expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated during acute KD (F =24.94,16.53,34.71,19.51;all P ＜ 0.05).Expression levels of PD-1,CD43 and its downstream molecules (SHP-2,PTEN,Vav) in CD19 + B cells were down-regulated during acute KD (F =6.43,5.57,19.52,10.37,11.37;all P ＜ 0.05),and restored remarkably after therapy (all P ＜ 0.05).Conclusion Insufficiency of IL-35 + Breg and its expression of IL-35 may be the important factors contributing to immunological dysfunction in KD.