Objective To explore mechanism of human bone mesenchymal stem cell(MSC)in treating patients with aplastic anemia(AA)in vitro.Methods MSCs were separated with Percoll(1.073 g/mL)and cultured in low glucose DMEM.T cells were harvested by using nylon column.MSCs of various concentrations were added to PHA induction T cell proliferation cultures with AA patients.The proliferation of T cell was measured by MTT method. The CD25(IL-2R)expression rates of CD~+_3 T cells was analyzed by flow cytometry .Results MSCs were planted in 96-well plates(2?10~4/well for group A,1?10~4/well for group B)and cocultured with T cell isolated from peripheral blood of AA patients. Peripheral blood T cell noncocultured with MSC acted as the control group.MSCs significantly inhibit PHA-induced T cell proliferation and the CD25 expression of CD~+_3 T cells in patients with AA(P

Objective To investigate the changes in the proportion of regulatory T (Treg) cells and in the levels of cytokines secreted by these cells in the peripheral blood in the patients with chronic myeloid leukemia (CML). Methods The enrolled subjects consisted of 30 CML patients who were newly diagnosed , 20 CML patients who were under the effective treatment of tyrosine kinase inhibitors (BCR-ABL 210 transcript ratio is below 10%) and 20 healthy donors whose age and sex were matched .Flow cytometry was used to detect CD4+CD25 high CD127 low /-Treg cells and CD4+ T cells.The enzyme linked immunosorbent assay was used to determine the plasma concentra -tions of interleukin-10(IL-10), transforming growth factor-β1(TGF-β1) and IL-35.Results The proportions of Treg cells in CD4+ T cells were similar among the three groups .As concerns the three kinds of Treg-associated cy-tokines, there were no significant differences in the plasma concentrations of IL -10 among the three groups.Howev-er, compared with the treatment group and the control group , the plasma concentrations of TGF -β1 and IL-35 in the newly diagnosed patients significantly increased (P <0.001), with no significant difference between the treat -ment group and the control group.Conclusion Though the proportion of Treg cells did not significantly change in the newly diagnosed patients, the plasma concentrations of TGF-β1 and IL-35 indeed significantly enhanced , sug-gesting the dysfunction of Treg cells in the newly diagnosed patients might be associated with the progression of dis -ease.Effective treatment of tyrosine kinase inhibitors could down -regulate the plasma levels of these cytokines to baseline, suggesting that monitoring these cytokines might evaluate the efficacy of therapy .

Objective To explore the alterations, relationship and clinical significance of CD4 +CD25 +CD127 low/ -
regulatory T cells ( Treg ) and lymphocyte subsets in peripheral blood of patients with acute myelocytic leukemia ( AML) . Methods The level of peripheral blood lymphocyte subsets and Treg of untreated AML patients and com-plete remission( CR) patients were tested by flow cytometry,and were compared with that of 30 normal controls. Re-sults The proportions of Treg were much higher in untreated AML patients and CR patients than in normal con-trols, while the mean proportion of Treg in untreated AML patients was higher than that in CR patients(P<0. 05). The proportions of NK( CD3 -CD16 +CD56 +) cells in untreated AML patients and CR patients were both decreased compared with normal controls,and the mean proportion of NK cells in untreated AML patients was lower than that in CR patients(P<0. 05). Compared with the normal controls,the proportions of CD3 +T cell, CD4 +T cell,and the ratio of CD4 +/CD8 + decreased in untreated AML patients ( P <0. 05 ) , but the proportions of CD8 +T cell was higher than in normal controls;the proportions of CD3 +T cell, CD4 + T cell, CD8 +T cell and the ratio of CD4 +/CD8 + in CR patients were close to the proportions in normal controls, but there was significant difference between CR patients and untreated AML patients(P<0. 05). Conclusion The increase of Treg, CD8 +T cell and decrease of NK cells, CD3 +T cell, CD4 +T cell, and the ratio of CD4 +/CD8 + in peripheral blood of patients with AML in-dicate that the immune function of patients with AML is depressed. Treg control the immune response of CD8 +T cells,at the same time inhibit the natural immune response of NK cells, playing a major role in the disorders of CD4 +T cells and CD8 +T cell balance,and closely relate with the development of AML. The immune treatment of patients with AML will be optimised by reducing the amount of Treg or removing the suppression function.

AIM:To study the effect of proprotein convertases (PCs) on the transforming growth factor (TGF) ?1-induced inhibition of HBV replication.METHODS:HepG2.2.15 cells cultured regularly were exposed to recombinant TGF?1 at concentration of 2 ?g/L or 5 ?g/L and/or PC inhibitor at concentration of 20 ?mol/L for 18 h.The total RNA and HBV core particle DNA were extracted from these cells,and PC mRNA and core-associated HBV DNA were detected by real-time PCR technique.RESULTS:The mRNA expression levels of 7 PCs in HepG2.2.15 cells were observed with various degrees.Recombinant TGF?1 significantly up-regulated the mRNA expression of all PCs except for the down-regulation of PC5 /6,though PC1 /3 and PC2 were up-regulated most obviously.Furin and PACE4 were the predominant PCs before and after TGF?1 exposure when the basic mRNA expression was taken into account.Further study showed that TGF?1-induced the inhibition of HBV replication was abrogated by PC inhibitor in HepG2.2.15 cells.CONCLUSION:TGF?1-induced the inhibition of HBV replication is mediated by the up-regulation of PCs,which might be of many implications in efficient interferences of TGF?1 on HBV replication.

Objecfive To detect the specific autoantibodies against platelet in idiopathic thrombocytopenic purpura (ITP) and to study the relationship between these autoantibodies and the severity of ITP as well as therapeutic effect.Methods Autoantibodies (GPⅡbⅢa and GPIb) against platelet glycoprotein was measured by a monoclonal antibody immobilization of platelet antigen assay (MAIPA) in 40 ITP patients.Results 10 patients had mono-specific antibodies to GP Ⅱ bⅢa and 6 patients had mono-specific antibodies to GPIbα.Another 20 patients had antibodies to both antigens and 4 patients had no detectable antibody to either platelet antigen.There Was negative correlation between the antibody against GPⅡbⅢa(b=-0.071,P<0.01),CPIbα(b=0.092,P<0.01) and platelet counts.The ratio of refractory cases in patients with antibodies to both antigens(8/20) was significantly higher than that in patients with mono-specific antibodies(1/16)(χ2=6.09,P<0.05).Conclusion The specific autoantibadies against platelet might be valuable for discrininafion of idiopathic thrombocytopenic purpura and non immune thrombocytopenia.The types of antibodies are related with severity of ITP and therapeutic effect.

Objective To analyze the clinical characteristics and prognostic factors in patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL),and to improve the diagnosis and treatment of PGI-DLBCL.Methods Retrospective analysis was conducted in 51 cases of PGI-DLBCL between January 2009 and August 2013.The data included clinical manifestations,pathological features,treatment regimens and prognosis.Results 51 patients included 31 males and 20 females,the range of ages was from 16 to 80 years old,median age was 48 years old.The major clinical presentation were abdominal pain,abdominal distension,abdominal mass,nausea and vomiting,abdominal mass.The occurrences in stomach,small intestine,colon,rectum and multiple involvement were 56.86 ％,29.41％,7.84 ％,1.90 ％ and 3.92 ％ respectively.The mass bigger than 10 cm was found in 13 cases (25.49 ％).47.06 ％ (24/51) of the cases belonged to the GCB subtype and 52.94 ％ (27/51) belonged to the non-GCB subtype.There was no significant impact of lymphoma cell origin,disease distribution (stomach or intestinal) and mass on prognosis of lymphoma treatment.The univariate analysis revealed that the patients with Lugano stage Ⅳ,increased level of serum lactate dehydrogenase (LDH),modified-international prognosis index (modified IPI) 3-5 and increased level of CA125 had poor prognosis (all P ＜ 0.05).There was no difference of survival rate between patients treated with rituximab plus chemotherapy and single CHOP like therapy.Surgery plus postoperative chemotherapy significantly improved survival of patients treated with simple chemotherapy (P ＞ 0.05).Conclusion The clinical Lugano stage,IPI score,increased LDH and CA125 are important prognostic factors of PGI-DLBCL.

By studying the behaviors of surfing in the Internet of the readers in our electronic reading room,we find a majority of readers are for amusement,only a minority of readers are to study and search data.Aiming at these problems,we put forward a series of measures and management countermeasures that guide readers to make good use of the electronic reading room and give some suggestions for our future work.

Objective To investigate the effect of leukemia-related genes on drug resistance in patients with acute myeloid leukemia (AML). Methods 74 patients with newly diagnosed AML were selected and 54 leukemia-associated genes of all patients were sequenced by second-generation gene sequencing. The gene with the highest mutation rate was further analyzed in association with resistance to several common chemotherapy medicines in in vitro drug sensitivity assays. In addition, in vitro drug resistance data were compared with the clinical data of patients. Results The TET2 gene was the most frequent mutation among 74 patients with newly diagnosed AML, with 11 positive patients. Among these 11 TET2 positive patients, 9 (81. 82％ ) were resistant to daunorubicin, while only 4 (6. 35％ ) out of 63 TET2 negative patients were resistant to daunorubicin. Besides, there was no significant difference between in vitro resistance rate to daunorubicin and the clinical data of patients. Conclusion TET2 gene mutation is associated with resistance to daunorubicin in AML patients, which may become an important indicator of the therapeutic efficacy of DA regimen.

Objective:To investigate the correlation between nutritional status and nosocomial infection in elderly patients with acute leukemia.Methods:A total of 202 elderly patients with acute leukemia hospitalized in the Department of Hematology of the First Affiliated Hospital of Anhui Medical University from June 2015 to December 2017 were randomly included as research objects.The nutritional status of patients was assessed by the Patient-Generated Subjective Global Assessment(PG-SGA), and the blood routine and biochemical indexes were compared among patients with different nutritional conditions.The correlations between clinical characteristics and PG-SGA score and between nutritional status and nosocomial infection were analyzed.Univariate analysis and multivariate Logistic regression analysis were used to evaluate the related factors for the nosocomial infection.Results:The hemoglobin and albumin were higher in patients with PG-SGA score of 0-8[(97.02±2.86)g/L and (39.78±0.50)g/L, respectively]than in patients with PG-SGA score ≥ 9[(83.02±3.28)g/L and (37.71±0.71)g/L]( P=0.003, 0.016). And C-reactive protein(CRP)was lower in patients with PG-SGA score of 0-8[(33.98±5.34)mg/L]than in patients with PG-SGA score ≥ 9[(58.82±8.36)mg/L]( P=0.015). There were significant differences in PG-SGA scores among patients with different age, disease stage, disease type and gastrointestinal reaction( t=-6.562, 3.292, 2.869 and 2.268, P=0.000, 0.001, 0.006 and 0.041). The PG-SGA score was positively correlated with the incidence of nosocomial infection( r=0.544, P=0.000). Logistic regression analysis showed that PG-SGA score was an independent risk factor( OR=2.11, 95% CI: 1.66-2.71, P=0.000). And albumin was a protective factor( OR=0.86, 95% CI: 0.77-0.93, P=0.000)for the nosocomial infection in elderly patients with acute leukemia. Conclusions:The nutritional status is closely related to the occurrence of nosocomial infection in elderly patients with acute leukemia.Elderly acute leukemia patients with malnutrition should be given adequate nutritional support as soon as possible to improve their nutritional status and improve prognosis.

OBJECTIVETo investigate the expression of metastasis-associated in colon cancer-1 (MACC-1) mediated by siRNA, and to study the effects of its downregulation on cell proliferation, cell cycle and invasion ability of cervical cancer SiHa cells.

METHODSMACC-1 siRNA and control siRNA were transfected into cervical cancer SiHa cells, and the expression of MACC-1 protein after transfection with MACC-1 siRNA was detected by Western blotting. The changes of cell proliferation, cell cycle and invasion ability of the SiHa cells were determined by CCK-8 kit, flow cytometry and Boyden chamber assay. The expressions of cell cycle- and invasion-related proteins were analyzed by Western blotting.

RESULTSCompared with the untreated group (0.317 ± 0.023) and control siRNA group (0.309 ± 0.021), the expression of MACC-1 protein was downregulated in the MACC1 siRNA group (0.041 ± 0.006) (P < 0.05), and its downregulation significantly suppressed the cell proliferation, altered the cell cycle distribution and reduced the cell invasion ability of the SiHa cells (P < 0.05). Compared with the untreated group (0.217 ± 0.025 and 0.215 ± 0.024) and the control siRNA group (0.222 ± 0.023 and 0.207 ± 0.027), the expression of cyclin D1 and Cdk2 proteins were significantly decreased in the MACC1 siRNA group (0.076 ± 0.010 and 0.039 ± 0.007) (P < 0.05). Compared with the untreated group (0.099 ± 0.007) and control siRNA group (0.105 ± 0.012), the expression of p21 protein was significantly increased in the MACC1 siRNA group (0.676 ± 0.044) (P < 0.05). The downregulation of MACC-1 expression also evoked a decrease of expressions of MMP-2 and MMP-9 proteins and an increase of E-cadherin protein expression (P < 0.05).

CONCLUSIONSMACC-1 downregulation-mediated inhibition of proliferation and decreased invasion ability of tumor cells may be closely associated with the alterations of expressions of cell cycle- and invasion-related proteins.

Cadherins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 2 ; metabolism ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; RNA, Small Interfering ; Transcription Factors ; metabolism ; Transfection ; Uterine Cervical Neoplasms ; metabolism