Objective:To systematically evaluate the efficacy and safety of demethylation drugs intreatment of myelodysplastic syndrome (MDS), and to provide reliable basis and guidance for clinical application of these drugs.Methods:The randomized controlled trials (RCT) of demethylation drugs for treatment of MDS published in the PubMed, Web of Science, Embase, Cochrane Library, Chinese Journal Full-text Database (CNKI), Chinese Biomedical Literature Database (CBM) and VIP database from January 2000 to December 2019 were searched by computer. RevMan 5.3 software was used for Meta-analysis of the efficacy and safety.Results:Seven RCT studies involving 1 172 patients were obtained. Meta-analysis showed that the complete remission rate ( OR = 6.26, 95% CI 1.74-22.49, P = 0.005), partial remission rate ( OR = 4.65, 95% CI 1.51-14.29, P = 0.007), overall response rate ( OR = 14.14, 95% CI 7.27-27.51, P < 0.01), hematology improves ( OR = 3.47, 95% CI 1.44-8.32, P = 0.005) in the demethylation drug treatment group were better than those in the best supportive treatment group. Meanwhile, the overall survival of patients in azacytidine group was improved ( HR = 0.62, 95% CI 0.50-0.77, P < 0.01). In terms of adverse reactions, demethylation drugs increased the incidence of neutropenic fever ( OR = 4.19, 95% CI 2.26-7.76, P < 0.01). However, there was no significant difference in the incidence of neutropenia, thrombocytopenia, anemia, infection, nausea, liver damage and fatigue between the two groups (all P > 0.05). Conclusions:The effect of demethylation drugs in treatment of MDS is obvious, the remission rate can be improved, and azacytidine can prolong the patients' survival, but demethylation drugs increase the incidence of neutropenia fever. In the clinical application of demethylation drugs, it is necessary to further carefully evaluate their clinical safety.

Objective To explore the influence of IL-2 pretreatment on splenic lymphocyte following mobilization with G-CSF, which may provide a new approach to attenuate acute graft-versus-host disease (GVHD). Methods Splenic cells and na(i)ve CD4+T cells from C57BL/6N mice receiving G-CSF-mobilized were pretreated with IL-2(50 U/ml) , and cocultured with the allogeneic antigens from BALB/c mice. The proliferation responses and the polarization of T cells were determined. C57BL/6N mice were randomly divided into 4 groups: G-CSF + IL-2 group, G-CSF group, IL-2 group , control group. Results Compared with the control group, IL-4 increased obviously while IFN-γ decreased significantly in the group of G-CSF + IL-2. The proliferation responses were also suppressed in vitro. Conclusion The proliferation responses of splenic cells and naive CD4 + T cells from C57BI/6N mice receiving G-CSF-mobilized to the allogeneic antigens were significantly abrogated by the pretreatment with IL-2, T cells were polarized toward the production of type-2 cytokines. The combination of G-CSF and IL-2 is potentially synergetic in the induction of T lymphocyte immune tolerance.

Objective To established the method of increasihg the proportion of T regulatory cells(Tr) to T effector cells(Te), which could suppresses allogeneic ahtigen reaction, by in vivo of glucocorti-coid (dexamethasone, DXM) combined with IL-2. Methods After combined treatment to male C57BL/6N mice (donor) with DXM(5 mg. kg-1· d-1 ) and IL-2 (300 000 IU · mouse-1·d-1) for 3 d, spleen mono-nuclear cells were made and were carried out by flow cytometry analysis. Using the spleen cells of BALB/c mice as aliogeneic antigen to stimulate the spleen cells of male C57BL/6N mice for 7 d after combined treat-ment of glucoeorticoid and IL-2, the reaction of cell proliferation was detected. Results After the treatrment of DXM and IL-2, CD4+ CD25+ Foxp3+ Tr cells in the spleen of C57BL/6N mice increased abviously. The ratio of CD25+ Foxp3+ Tr to CD4+ T cells was 24.22%±7.60% in the group of DXM combined with IL-2, while the control group was 4.02% ±0. 84% ( P =0. 01 ). Compared with the control group (0. 14±0.01 ), the ratio of Tr to Te increased obviously in the group of DXM combined with IL-2 (0.43±0. 15 ) ( P = 0.01 ). The group of DXM combined with IL-2 also expressed more glucocorticoid-induced tumor necrosis factor receptor(GITR) and alloreaction was suppressed/n vitro obviously ( P < 0. 05 ). Conclusion DXM amplifies IL-2 induced the proportion of Tr to Te, and suppresses the cell proliferation stimulated by alloge-neic antigen.

Purpose:To evaluate the effect of Auto-PBSCT combined with long-time low-dose interleukin-2 (after transplant) for increasing the rate of 5-year disease-free survival of lymphoid malignancies (ALL and NHL).Methods:U- sing chemotherapy combined with low dose G-CSF to mobilize peripheral blood stem cells,we carried out auto-PBSCT in lymphoid malignancies (7 cases CR_1 ALL、1 case CR_2 ALL、3 cases CR_1 NHL、5 cases refractory NHL).After transplant, 15 cases used low dose interleukin-2 at 5.10~5U/d for one year or longer.Results:7 cases CR_1 ALL are still alive,the aver- age disease-free survival is 73 months.Median disease-free survival is 51.5 months.2 of 3 cases CR_1 NHL are still alive. The 6 years disease-free survival in 10 cases CR_1 lymphoid malignancies(CR_1 NHL and CR_1 ALL) is 0.90?0.11,the aver- age disease-free survival is 66.8 months.In another PR (refractory) group of NHL and ALL (CR_2),3 years disease-free survival was 0.33?0.18,and average survival time was 20.66 months,but a chemotherapy-sensitive NHL is still alive (63 months),a case of lymphoblast cell NHL also survived for 46 months and died after relapse.Conclusions:Auto-PBSCT combined with long-turm low dose IL-2(after transplant) could effectively increase the rate of 5-year disease-free survival in CR_1 lymphoid malignancies(ALL and NHL).

Objective To further explore the relationship between increasing genetic material and clinical manifestation of partial trisomy 5p .Methods G‐banding karyotypes of peripheral blood lymphocytes in the patient and his parents ,and at the same time to summary the partial trisomy 5p clinical performance .Results patient ,46 ,XX ,der(6)t(5 ;6)(p13;q25) mat ;partial trisomy for 5p13→pter resulting from the balanced translocation of the mother .Mother:46 ,XX ,t(5;6)(p13 ;q25);carrier of a balanced 5/6 translocation .Father :46 ,XY .Conclusion The phenotype of trisomy 5p may be associated with express and function of gene at spe‐cial chromosome region .

Objective To investigate the influence of Herba Epimedium, a kidney-strengthening Chinese herb, on the regenerate ossification of nano-hydroxyapatite/collagen/ polylactic acid (nHAC/PLA) in the repair of rabbit mandibular defect. Methods Sixteen rabbit models with 10 mm × 8 mm subperiosteal mandibular defect were established. The rabbits were randomly divided into 2 groups, namely Herba Epimedium group and control group. In both groups, the left mandibular defect region (short for left side) was not filled with any material, while the right mandibular defect region ( short for right side) was filled with nHAC/PLA. On week 2, 4, 8, 12 after modeling, rabbits in each group were killed, and then the macro-view, imageology and histology of the bilateral mandibles were observed. The data of the regenerate bone areas were calculated and analyzed with analysis of variance (ANOVA) . Results The imageological results showed bone mineral density in the right side of Herba Epimedium group and the control group, and in the left side of Herba Epimedium group were higher than those in the left side of the control group on week 2, 4, 8, 12 after modeling. The histological results showed that the regenerate ossification in the right side of Herba Epimedium group and the control group was superior to that in the left side of the corresponding groups, with more regenerative bone and vessels. In the right side of Herba Epimedium group and the control group, amount of bone-like tissues were grown into nHAC/PLA, and the maturity degree of bone tissue was increased with the prolongation of time. And the changes were more obvious in the right side of Herba Epimedium group, showing as the remaining of nHAC/PLA having been surrounded and replaced by regenerative bone tissue. Regenerative bone area was increased in both sides of Herba Epimedium group, which differed from the same side of the control group (P<0.05) . The differences of the regenerative bone area were also significant between the left side and right side of the same group (P<0.05) . Conclusion Herba Epimedium can effectively accelerate the repair of mandibular defect, and nHAC/PLA is of good biocompatibility, osteoinduction and bone conduction, and can be expected for bone defect repairing in clinic.

Objective To valuate the applicability of multiplex ligation‐dependent probe amplication (MLPA ) for detection of chromosomal unbalance .Methods Aneuploid and subtelomeric MLPA technology were used to analyze 6 samples with chromosom‐al unbalances ,confirmed by karyotype analysis .Results All samples were identified to have abnormal signal of corresponding probes .The extent of unbalances contained subtelomeric region .Conclusion MLPA technology could have important role in diag‐nosis of chromosomal unbalance ,which could complement conventional karyotype analysis .

Objective To identify gene mutations and explore the molecular mechanism of a pedigree with inherited coagulation F Ⅶ deficiency. Methods The levels of F Ⅶ: Ag in the proband and other family members were measured by ELISA assay. The values of PT, F Ⅶ: C and other coagulant parameters were determined by one-stage clotting for laboratory phenotype diagnosis. All the exons,exon-intron boundaries and 5',3' untranslated sequences of F7 gene were amplified by direct sequencing. The detected mutations were further confirmed by sequencing the other stand. The CLC Protein Workbench software was used to analyze the species conservation of the mutated site and the protein secondary structure. 100 healthy individuals were selected to exclude gene polymorphism. Results PT, FⅦ∶C and FⅦ: Ag in the proband and his sister were abnormal, which were 36. 3 s, 5.0%, 40. 7% and 33.4 s,5. 0%, 37.4%, respectively. Both PT and FⅦ∶C in the proband's father, mother, daughter and niece were slightly abnormal, which were 14.9 s, 14. 6 s, 15.5 s, 14. 6 s and 70%, 85%, 59%, 79%, respectively.The heterozygous mutations c. 784T ＞ C and c. 964T ＞ G in exon 8 of F7 gene were found in the proband,resulting in the substitutions of Ser269Pro and Cys329Gly respectively. Compound heterozygous mutations c. 784T ＞ C and c. 964T ＞ G were found in the proband's sister. The proband's mother was heterozygous for c. 784T ＞ C. His father, daughter and niece were heterozygous for c. 964T ＞ G. The protein biological characteristics analysis revealed that the Cys329Gly caused the change of spatial configuration, and Ser269Pro led to the change of amino acid polarity and hydrophobicity. Conclusion Compound heterozygous mutations of Cys329Gly and Ser269 Pro in F7 gene may be the underlying molecule mechanism of FⅦ deficiency in this pedigree.

Objective To explore the polarization and proliferation of naive CD4+TT cells after preincubation with interleukin-2. Methods The expression of suppressor of cytokine signal (SOCS)-3 was detected by the real-time PCR in the naive T lymphocytes of DO11.10 TCR transgenic and C57BL/6N mice after they were preincubated by 50 U/ml of IL-2. The naive CD4+T T lymphocytes preincubated for 4 h by IL-2 were stimulated with ovalbumin (OVA) and inactivated BALB/c spleen cells, respectively. After 14 d, IL-12Rβ1, IL-12β2 and cytoplasm IL-4 (cyIL-4) were detected by flow cytometry. Results SOCS-3 in the naive CD4+T lymphocytes of DO11. 10 TCR transgenic and C57BL/6N mice reached the peak after preincu- bated by IL-2 for 6 h. The polarization to TH1 and the proliferation ability of naive CD4+ T lymphocytes stimulated with specific antigen and allogeneic antigen were inhibited conspicuously during the time of the peak of SOCS-3. Conclusion The expression of SOCS-3 in naive CD4+T lymphocytes can be upregulated by IL-2. The upregulation of SOCS-3 can suppress the polarization to TH1and the proliferation ability of naive CD4+T lymphocytes stimulated by specific antigen and allogeneic antigen.

Objective To investigate the variety and clinical value of the neutrophil volume and cytoplasm-nucleus complex in patients with bacterial infection, cardiovascular or cerebrovascular accident and major surgery operation. Methods 125 patients with bacterial infection, 64 patients with acute cardiovascular or cerebrovascular accident, 66 patients after major surgery operation and 69 normal subjects were selected in the study. Total WBC counts (WBC), percentage of neutrophils (NE), and the VCS parameters of neutrephils including the mean channels of cell volume ( NEV), conductivity ( NEC), light scatter(NES) and the SD of these parameters( NEVSD, NECSD, NESSD)were measured by automatic blood cell analyzing instrument. The sensitivity and specificity of the WBC, NE and the VCS parameters of neutrophils were analyzed with receive operating characteristic (ROC) curve. Results The levels of NEV, NES and NEVSD in acute bacterial infection group were 154.3 ± 15.2, 135.7 ± 9.9, 26.8 ± 4.2 respectively. The levels of NEV, NES and NEVSD in post major surgery operation group were 147.2±8.9, 141.5 ± 7.7, 23.0 ± 2. 8 respectively. The levels of NEV, NES and NEVSD in acute cardiovascular or cerebruvascular accident group were 144.9 ± 5. 2, 146.0 ±5.0, 19. 6±1.6 respectively. The levels of NEV, NES and NEVSD in healthy control group were 139.7±4.6, 145.0±3.8, 18.2±1.3 respectively. The differences of these parameters among these groups had statistical significance ( F = 17. 650, 38. 122, 54. 604,P<0. 05). And the changes of NEV, NES and NEVSD in bacterial infection group were most obvious among those three groups. The levels of NEV, NES and NEVSD were 146.5±9.5, 144.3 ± 9.4, 21.3 ± 3.3 respectively in stress diseases groups which included acute cardiovascular or cerebrovascular accident group and post major surgery operation group. The differences of these parameters between stress diseases group and acute bacterial infection group had statistical significance ( t = - 2.840, 7.533, - 8.999,P<0.01). The areas under the ROC curve of NEVSD, NEC, NES and NECSD were 0.893, 0. 845, 0. 833 and 0. 849 respectively. The sensitivity of 83. 3% and specificity of 82. 0% could be achieved by selecting the cut-off equal to or greater than 24. 0. Conclusions The variety of neutrophil volume, nuclear size and cytoplasmic granularity changed obviously in patients with acute bacterial infection and common stress diseases, and the variety in acute bacterial infection is more obvious than that in common stress diseases. The sensitivity and specificity of VCS parameters of neutrophils are higher than those of WBC or NE for predicting infection, and the NEVSD is the most predictable indicator of acute bacterial infection.