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1.
Article in Chinese | WPRIM | ID: wpr-829048

ABSTRACT

OBJECTIVE@#To study the expression of multiple negative costimulatory molecules on peripheral blood T cells in patients with acute myeloid leukemia (AML) and its affection on prognosis.@*METHODS@#The peripheral blood samples from patients with newly diagnosed AML, complete remission (CR), and no-remission (NR) were collected, the expression levels PD-1、VISTA and TIM-3 in CD4 and CD8 T cells were detected by flow cytometry , and the clinical data of patients were analyzed.@*RESULTS@#The expression levels of PD-1、VISTA and TIM-3 of CD4 and CD8 T cells in the newly diagnosed AML patients were significantly higher than those in control group (P<0.05). The expression levels of PD-1、TIM-3 and VISTA of CD4 and CD8 T cells in the CR group were significantly lower than those in newly diagnosed and the NR group (P<0.05). The TIM-3 expression level positively correlated with VISTA expression level of CD4 and CD8 T cells in newly diagnosed AML patients (r=0.85 and 0.73). The VISTA and PD-1 expression level of CD4 T cells in newly diagnosed AML, NR after first induction chemotherapy and high risk patients significantly increased (P<0.05), the TIM-3 expression level of CD8 T cells in high risk group significantly increased (P<0.05), and the VISTA expression level of CD8 T cells in CBFβ-MYH11 mutation-positive group significantly decreased (P<0.05).@*CONCLUSION@#The expression of PD-1、TIM-3 and VISTA in AML peripheral blood T cells may be involved in the immune escape of AML and can be the targets of treatment for acute myeloid leukemia patients.


Subject(s)
B7 Antigens , CD8-Positive T-Lymphocytes , Flow Cytometry , Hepatitis A Virus Cellular Receptor 2 , Humans , Leukemia, Myeloid, Acute , Programmed Cell Death 1 Receptor
2.
Journal of Experimental Hematology ; (6): 1228-1233, 2020.
Article in Chinese | WPRIM | ID: wpr-827135

ABSTRACT

OBJECTIVE@#To investigate the expression and clinical significant of VCAN and its related molecules in patients with MM.@*METHODS@#Ficoll density gradient centrifugation method was used to speared the bone marrow mononuclear cell in 25 cases of MM before and after treatment, the relative mRNA expression of VCAN and their related molecules (FAK, FN, MK, and HAS) in bone marrow was detected by real-time quantitative PCR, and their protein expression was determined by Western bolt.@*RESULTS@#The expression of VCAN, FK and FN in the effective group after treatment was significantly lower than that before treatment (P<0.05), however, the expression of MK and HAS showed no statistically significantly different before and after treatment (P<0.05). The expression of VCAN of patients in non remission group was significantly higher than that in control group (P<0.05). The expression of FAK and FN of patients in no remission group was significant increased as compared with the patients in newly diagnosed group (P<0.05). The relative expression of VCAN mRNA in the patients at 3rd stage was significantly higher than those at the 1st stage (P<0.05) and control group but showed no significant difference to the patients at 2nd stage (P<0.05). The expression of VCAN and its related proteins (FAK, MK, FN) showed positively correlation in bone marrow mononuclear cells of MM patients (P<0.05). The correlation between VCAN and HAS was not statistically significant (r=0.259,P>0.05). Survival analysis showed that the relative expression of VCAN mRNA was associated with OS (P=0.049) and PFS (P=0.041) in MM patients.@*CONCLUSION@#VCAN and its related molecules are highly expressed in MM patients; VCAN may act as potential biomarker in the development of multiple myeloma.


Subject(s)
Bone Marrow , Humans , Multiple Myeloma , RNA, Messenger , Versicans
3.
Article in Chinese | WPRIM | ID: wpr-827196

ABSTRACT

OBJECTIVE@#To explore the expression and clinical significance of EZH2 in DLBCL patients accompanied by HBV infection.@*METHODS@#The clinicopathological data of 59 patients with DLBCL accompanied by HBV infection in our hospital from February 2015 to October 2017 were analyzed retrospectively. The patients were divided into HBV negative and HBV positive groups by serological testing before surgery. The expression of EZH2 was detected by immumohistochemical staining, and the clinicopathological characteristics and survival were analyzed and compared between these two groups.@*RESULTS@#There were 30 patients (50.8%) in the HBV negative group and 29 patients (49.2%)in the HBV positive group. The differences of age, LDH level and IPI score between two groups were statistically significant (P<0.05). The expression of EZH2 in HBV- positive group was significantly higher than that in the HBV- negative group (P<0.05), where the expression of EZH2 correlated with the expression of the BCL-6 (r=0.282, P<0.05), especially in the GCB-DLBCL (r=0.549, P<0.05). PFS was not significantly different between two groups of HBV (P>0.05), while the PFS in the R-CHOP regimen group was higher than that in the CHOP regimen group (P<0.05). COX multivariate analysis showed that both the chemotherapy regimen without R and the increased level of LDH were the risk factors affecting the prognosis of DLBCL patients (P<0.05).@*CONCLUSION@#EZH2 highly expresses in HBV positive group, suggesting that the significance of EZH2 in DLBCL with HBV infection is worth further explore.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols , Cyclophosphamide , Doxorubicin , Enhancer of Zeste Homolog 2 Protein , Genetics , Hepatitis B , Hepatitis B virus , Humans , Lymphoma, Large B-Cell, Diffuse , Genetics , Prognosis , Retrospective Studies , Rituximab , Vincristine
4.
Journal of Experimental Hematology ; (6): 1190-1195, 2019.
Article in Chinese | WPRIM | ID: wpr-775743

ABSTRACT

OBJECTIVE@#To investigate the relationship of cytogenetic features, clinical characteristics and prognosis in patients with myelodysplastic syndrome.@*METHODS@#The clinical characteristics and prognosis of 236 patients with MDS admitted to the Affiliated Hospital of Xuzhou Medical University from January 2013 to September 2017 were analyzed retrospectively, the follow-up observation and correlation analysis were performed.@*RESULTS@#There were 33 cases of refractory cytopenia with unilateral dysplasia (RCUD), 8 cases of refractory anemia with ring-shaped iron granulocytes (RARS), 70 cases of refractory cytopenia with multiple dysplasia (RCMD), 23 cases of refractory anemia (RA), 46 cases of refractory anemia with excessive blasts (RAEB-1), 48 cases of (RAEB-2), MDS-U 2 cases, simple del(5q) 6 cases. The detection analysis showed that the chromosome abnormality rate and complex chromosome abnormality rate in RAEB group (RAEB-1 + RAEB-2) and in non-RAEB group (RARS+RCMD+RCUD+RA) were 48.94% vs 43.94%, and 18.09% vs 12.69%, respectively, which were no statistically different. The grouping according to IPSS-R and IPSS showed that the chromosome abnormality rate gradually increased along with enhancement of risk stratifi-cation (P<0.05). The cytogenetic characteristics analysis showed that a total of 108 cases had chromosomal abnormalities, the detection rate was 45.76%, Out of 108 cases, 36 cases had complicated karyotypes, accounted for 15.25% of all patients. The types of chromosomal abnormalities mainly include numbers, structural abnormalities and mixed abnormalities. The chromosome abnormality with the highest detection rate was +8, accounted for 30.56% (33/100) of patients with chromosome abnormalities; followed by -7/7q-, del(5q), del(20q), etc. -7/7q-chromosome abnormalities accounted for 29.63% of all karyotypic abnormalities (including -7/7q-chromosome abnormalities alone and other chromosome abnormalities). The median age of the patients with MDS was 61 (13-88) years old, and the male-female ratio was 1.36∶1. Analysis of blood cell characteristics showed that the three lines were reduced or increased to varying degrees. The median WBC count was 2.8 (0.3-267.1)×10/L, the median Hb level was 69 (20-156) g/L, and the median Plt count was 55 (2-1733)×10/L. The 1 year OS rate in 32 cases of chromosome 7 abnormality and 128 cases of normal chromosome was 25% and 44.53%, respectively, the difference was statistically significant (χ=4.05, P<0.05) .@*CONCLUSION@#Chromosome karyotype is an independent factor affecting the prognosis of patients with MDS. It is important for the diagnosis, treatment and prognosis evalnation of patients with MDS. The overall prognosis of patients with abnormal chromosome 7 is poor. .


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Chromosome Aberrations , Cytogenetics , Female , Humans , Karyotyping , Male , Middle Aged , Myelodysplastic Syndromes , Prognosis , Retrospective Studies , Young Adult
5.
Journal of Experimental Hematology ; (6): 1311-1315, 2019.
Article in Chinese | WPRIM | ID: wpr-775723

ABSTRACT

Abstract  Double-hit lymphoma (DHL) is a high-grade B-cell lymphoma with MYC and BCL-2/BCL-6 rearrangement, which is a invasive disease with a poor prognosis. FISH is the most important diagnostic method. There is no standard protocol for this disease yet. New therapeutic approaches including targeted therapy,checkpoint inhibitors and chimeric antigen receptor T-cell therapy are changing the paradigm of treatment for DHL. This review summarizes new developments in diagnosis and treatment of double-hit lymphoma.


Subject(s)
Genetic Predisposition to Disease , Humans , Immunotherapy, Adoptive , Lymphoma, B-Cell , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins c-bcl-6 , Proto-Oncogene Proteins c-myc
6.
Journal of Experimental Hematology ; (6): 1602-1606, 2019.
Article in Chinese | WPRIM | ID: wpr-775678

ABSTRACT

OBJECTIVE@#To investigatc the curative efficacy of low dose rituximab for glucocorticoid ineffective on dependent ITP patients and its relation with sensitivity to glucocorticoid so as to provide reference basis for rational use of drugs in clinical treatmant.@*METHODS@#Seventy-ninth ITP patients enrolled in this study included the glucocorticoid-ineffective patients (19 cases) and glucocorticoid-dependent patients (60 cases). All ITP patients were treated with regimen consisted of high dose dexamethasone plus low dose rituximab (dexal-methasone 40 mg/d for 4 days per os, ritaximab 100 mg by intravenous infusion at D7, 14, 21 and 28 respectively). The patients after treatment were followed-up for 12 month, and the relation of patients sensitivity to glucocorticoid with therapentic response of rituximab was analyzed. The changes of Treg cell ratio and BAFF, IL-2 and sCD40L levels before and after treatment were detected by flow cytometry and ELISA respectively.@*RESULTS@#The overall response rate (ORR) of patients treated with above- mentioned regemen at 1, 3, 6 and 12 months after treatment was 79.7% (63/79), 69.6% (55/79), 63.3% (50/79) and 60.8% (48/79) respectivcly, out of which the ORR of glucocorticoid ineffective and glucocorticoid-dependent ITP patients treated with above-mentioned regimen at 1, 3, 6 and 12 months after treatment was 47.4% (9/19) vs 90.0% (54/60), 36.8% (7/19) vs 80.0% (48/60), 21.1% (4/19) vs 76.7% (46/60), 21.1% (4/19) vs 73.3% (44/60), and the difference between 2 groups was statistically significant. The detection of T reg cell showed that the T reg cell ratio in glucocorticoid- ineffective and dependent patients at 1, 3, 6 and 12 months after treatment was (1.70±0.43)% vs (3.47±0.72)%, (1.66±0.33)% vs (4.29±0.91)%, (1.71±0.37)% vs (4.44±0.97)%, (3.36±0.54)% vs (4.29±1.04)%, respectively. The detection of cytokines showed that the levels of BAFF, IL-2 and sCD40L in plasma of glucocorticoid-dependent patients at 1 month after treatment significanlly decreased (P<0.05), the levels of BAFF, IL-2 and sCD40L in plasma of glucocorticoid-ineffective patients although decreased at 1 mouth after treatment, but there was no statistical difference as compared with glucocosticoid-depenment patients.@*CONCLUSION@#The treatment of glucocorticoid-dependent ITP patients with rituximab is more effective. The regulatory effect of rituximab on the T-reg cells, BAFF, IL-2 and sCD40L may be one of its mechanisms.


Subject(s)
Dexamethasone , Glucocorticoids , Humans , Inosine Triphosphate , Purpura, Thrombocytopenic, Idiopathic , Drug Therapy , Rituximab , Therapeutic Uses
7.
Journal of Experimental Hematology ; (6): 1329-1333, 2016.
Article in Chinese | WPRIM | ID: wpr-332693

ABSTRACT

<p><b>OBJECTIVE</b>To establish a BALB/c nude mouse model with the huamanized chronic myeloid leukemia (CML) for the study of human CML.</p><p><b>METHODS</b>The BALB/c nude mice aged 4 weeks pretreated by splenectomy, the cyclophosphamide intraperitoneal injection and sublethal irradiation (SLI) were transplanted intravenously with bone marrow mononuclear cells from CML patients. The SLI-pretreated nude mice were divided into 2 groups: group A, in which the nude mice were injected with 0.3 ml PBS; group B, in which the nude mice were infused intravenously with 4.5×10mononuclear cells from CML patients. Then the changes of body weight and appetite were observed, the hemogram and cell morphology were determined, the expressions of human CD13 and CD45 were detected by flow cytometry, the pathologic analysis of bone, liver and intestine were performed by biopsy, and the BCR/ABL fusion gene was detected by RT-PCR.</p><p><b>RESULTS</b>The mice in group B displayed weakness, auantic, less foodintake and instabiligy of gait as time want on. The average survival time was 46.2±4.2 d (45-57 d). On the third week, the CD13CD45cells accounted for 0.56±0.05% and 2.56±0.36% respectively in group A and B. While on the sixth week, the CD13CD45cells accounted for 0.44±0.07% and 4.97±0.43% in A and B groups respectively, these results showed that cell count in B group was significantly higher than that in A group(P<0.05). Pathological examination showed that the leukemic cells were found in bone marrow of group B. The BCR/ABL fusion gene could be detected in bone marrow.</p><p><b>CONCLUSION</b>BALB/c nude mouse model with huamanized chronic myeloid leukemia(CML) model has been established by pretreating mice with SLI. The survival time of mice in this model has been long, and the cost to establish the model is low.</p>

8.
Chinese Journal of Hematology ; (12): 478-481, 2013.
Article in Chinese | WPRIM | ID: wpr-235422

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the change of CD4⁺CD25(high)CD127(low) regulatory T cells (Tregs) percentage in patients with primary immune thrombocytopenia (ITP) treated by different methods.</p><p><b>METHODS</b>One hundred and thirty-eight newly diagnosed adult ITP patients (57 male, median age 40 years, range 18-70 years) were enrolled in this study, who were randomly separated into three regiment groups, namely prednisolone (PSL, 1.5 mg/kg for 2-4 weeks and subsequently stepwise reduction) group enrolled 49 patients, dexamethasone [(one course of high-dose dexamethasone (HDD) 40 mg/day, d1-4] 45 patients, and rituximab plus HDD (rituximab 100 mg on days 7, 14, 21, 28 and HDD) group 44 patients. Peripheral blood was taken in ITP patients of each group before treatment, 14 d and 28 d after treatment. The percentages of CD4⁺CD25(high)CD127(low) Tregs in 30 healthy controls and 138 patients were analyzed by flow cytometry.</p><p><b>RESULTS</b>Overall response (OR) rates of PSL, HDD and R+HDD groups at day 28 were 69.4%, 66.7% and 79.5% respectively with no difference. After the following 12 months, sustained response (SR) was more pronounced in R+HDD group compared to the other two groups (R+HDD vs PSL: 66.7% vs 37.8%, P<0.05; R+HDD vs HDD: 66.7% vs 22.7%, P<0.05). The percentage of CD4⁺CD25(high)CD127(low) Tregs in peripheral blood of ITP patients [(1.67±0.70)%] was significantly lower than in healthy control group; After treatment, the percentages of Tregs in peripheral blood of patients both at day 14 and 28 in R+HDD group remarkably decreased compared with before treatment [(4.28±1.09)% vs (1.68±0.68)%, P<0.05; (4.44±0.63)% vs (1.68±0.68)%]. The percentages of Tregs at day 14 in both other two groups decreased notably compared with before treatment. But the Tregs levels measured at day 28 in PSL and HDD groups were similar with before treatment.</p><p><b>CONCLUSION</b>The percentage of CD4⁺CD25(high)CD127(low) Tregs in peripheral blood of ITP patients was lower than healthy individual. The higher SR of patients treated by R+HDD was related to its ability to up-regulate the percentage of CD4⁺CD25(high)CD127(low) Tregs.</p>


Subject(s)
Adolescent , Adult , Aged , Antibodies, Monoclonal, Murine-Derived , Therapeutic Uses , Dexamethasone , Therapeutic Uses , Female , Humans , Male , Middle Aged , Prednisolone , Therapeutic Uses , Rituximab , T-Lymphocytes, Regulatory , Thrombocytopenia , Blood , Drug Therapy , Allergy and Immunology , Young Adult
9.
Chinese Journal of Hematology ; (12): 782-787, 2013.
Article in Chinese | WPRIM | ID: wpr-272114

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of immature dendritic cells (imDC) expressing chemokine receptor-7 (CCR7) on acute graft-versus-host disease (aGVHD) in allogeneic bone marrow transposed (allo-BMT) mouse model.</p><p><b>METHODS</b>We constructed the lentiviral vectors carrying mouse CCR7 gene and infect imDC effectively in vitro. GVHD model was established with C57BL/6(H-2b) donor mice and BALB/c (H-2d) recipient mice. After irradiation, recipients were injected with donor bone marrow and spleen cells along with CCR7-modified dendritic cells. Mice were randomized into irradiation, transplant control, pXZ9-imDC (empty vector control) and CCR7-imDC groups. Survival, GVHD score, histopathological analysis and plasma levels of inflammatory cytokines were observed.</p><p><b>RESULTS</b>The mean survival in irradiation, transplantation, pXZ9-imDC and CCR7-imDC groups were (8.20±1.48)d, (12.20±2.78)d, (20.70±6.01)d and (27.5±7.55)d respectively. The survival in CCR7- imDC group was significantly improved compared with other groups (P<0.05). GVHD scores in transplantation, pXZ9-imDC and CCR7-imDC groups were (6.90±1.66), (5.60±0.97) and (4.10±1.79) respectively. CCR7-imDC group had significantly lower GVHD score and minor tissue damages shown by histopathological analysis than the other groups. Plasma IFN-γ level increased and reached the peak at +10 day in transplant group, while it gradually decreased in pXZ9-imDC and CCR7-imDC groups, and then reached the nadir at +20 day post-allo-BMT, with the lowest level in CCR7-imDC group (P<0.01). Plasma IL-4 decreased in transplant group, while it gradually increased in pXZ9-imDC and CCR7-imDC groups and reached the highest level at + 10 day in CCR7- imDC group (P<0.01). The 95%-100% of H-2b positive cells in recipient mice on + 30 day post-allo-BMT demonstrated the complete donor- type implantation.</p><p><b>CONCLUSION</b>Genetically modified immature DC by CCR7 gene could alleviate damages by GVHD and prolong survival of recipient mice after allo-BMT.</p>


Subject(s)
Animals , Bone Marrow Transplantation , Dendritic Cells , Cell Biology , Genetic Vectors , Graft vs Host Disease , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Receptors, CCR7 , Genetics , Transplantation, Homologous
10.
Journal of Experimental Hematology ; (6): 1546-1551, 2013.
Article in Chinese | WPRIM | ID: wpr-264978

ABSTRACT

This study was aimed to investigate the effect of Wnt/β-catenin signaling pathway on the biologic behavior of mouse bone marrow mesenchymal stem cells (mBM-MSC) by constructing a RNAi lentiviral vector specific to β-catenin. Three pairs of shRNA coding sequences directed against different sites of β-catenin mRNA were designed and were linked into lentiviral vector plasmid PLB for constructing the PLB-β-catenin/shRNA1, PLB-β-catenin/shRNA2 and PLB-β-catenin/ shRNA3. Those plasmids and lentiviral packaging plasmids were co-transfected into the packaging cells 293FT, then the virus particles were collected and the viral titer was assayed after concentration, and these viral particles were infected to MSC. The flow cytometry was used to sort GFP (+) cells, Western blot and RT-PCR were used to verify the inhibitory effect of those cells on expression of β-catenin gene in cells, CCK-8 method was used to detect the cell proliferation level, Annexin-V/7-AAD was used to determine the cell apoptosis after interference, the cell scratch and transwell tests were used to detect the migration capability of MSC. The results showed that the efficient inhibition of β-catenin mRNA and protein expression, and the suppression of MSC proliferation were observed in group of PLB-β-catenin/shRNA2 (interference group), while there was no significant changes of MSC proliferation between negative group (PLB group) and the normal group (control group). The flow cytometric detection indicated that after induced by serum starvation for 72 h, the apoptosis of MSC increased in interference group, but there was no difference between PLB and control groups (P > 0.05). The cell scratch and transwell tests demonstrated that the migration capability of MSC in interference group decreased significantly, while the migration capability of MSC in control group was not changed obviously. It is concluded that the constructed specific RNAi lentivirus can effectively inhibit the expression of β-catenin gene, decrease expression level of β-catenin mRNA and protein. The Wnt/β-catenin signaling pathway plays an important role in biological behavior of BM-MSC.


Subject(s)
Animals , Apoptosis , Bone Marrow Cells , Cell Biology , Cell Line, Tumor , Cell Movement , Genetic Vectors , Lentivirus , Genetics , Mesenchymal Stem Cells , Cell Biology , Mice , RNA Interference , RNA, Small Interfering , Wnt Signaling Pathway , beta Catenin , Metabolism
11.
Chinese Journal of Hematology ; (12): 362-365, 2012.
Article in Chinese | WPRIM | ID: wpr-359483

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of thrombopoietin (TPO) on platelet engraftment in hematological malignancies patients after allogeneic haematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>One hundred and twenty patients were enrolled in a multicenter, open-label, randomized, controlled clinical trial, and were randomized into 4 treatment groups following allo-HSCT. Group A was the control arm without TPO, while group B, C and D were trial arms with received 300 U×kg(-1)×d(-1) of TPO starting from day +1, +4 and +7, respectively. A total of 89 cases were evaluated, of which 22 cases in group A, 23 in group B, 20 in group C and 24 in group D. Efficacy evaluation (the time of platelet engraftment, the number of platelet transfusion) and safety evaluation \[adverse events, routine blood tests, liver and renal function, coagulation function and occurrence of graft-versus-host disease (GVHD)\] were observed.</p><p><b>RESULTS</b>The median platelet engraftment time in experimental groups (groups B, C and D) were on day (13.17 ± 2.89), day (12.15 ± 2.08), day (12.33 ± 1.76), respectively, and that in control group was on day (14.82 ± 5.05). There was statistically significant difference between two groups (P = 0.029), There were no statistically significant difference in the average amount of platelet transfusion, platelet engraftment time, and platelet nadir value among the 3 experimental groups. No significant adverse events were observed in experimental groups.</p><p><b>CONCLUSIONS</b>TPO administration following allo-HSCT for patients with hematologic malignancies appears to shorten platelet engraftment time. TPO given starting from day +7 is effective and safe.</p>


Subject(s)
Adolescent , Adult , Blood Platelets , Child , Female , Hematologic Neoplasms , General Surgery , Hematopoietic Stem Cell Transplantation , Humans , Male , Middle Aged , Platelet Transfusion , Methods , Thrombopoietin , Therapeutic Uses , Transplantation, Homologous , Young Adult
12.
Chinese Journal of Hematology ; (12): 88-93, 2012.
Article in Chinese | WPRIM | ID: wpr-345934

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of immature dendritic cells (inDC) genetically modified to express sTNFR I on acute graft-versus-host disease (aGVHD) and the graft-versus-leukemia (GVL) effect ofter allogeneic bone marrow transplantation (allo-BMT) in leukemic mice and its mechanism.</p><p><b>METHODS</b>An EL4 leukemia allo-BMT model was established with the BALB/c (H-2d) donor mice (DM)and C57BL/6 (H-2b) recipient mice (RM). The RM received DM bone marrow (BM) cells at a 1:1 ratio with spleen cells intravenously via tail vein at 4 h after TBI. Fifty DM were separated randomly into five groups: (1) Group A: total body irradiation (TBI) group, (2) Group B: lymphoma cell leukemia group, (3) Group C: allo-BMT group, (4) Group D: pXZ9-DC group, (5) Group E: sTNFR I-DC group. Acute GVHD scores, incidence of leukemic cell infiltration, histopathological analysis, survival rate, and survival rate of the recipients were estimated after allo-BMT. Enzyme-linked immunosorbent assay (ELISA) method was used to detect cytokines (INF-gamma and IL-4 ) production. Flow cytometry (FCM) analysis was used to detect allogeneic chimerism.</p><p><b>RESULTS</b>(1) The mice in group A and group B all died of the BM failure and lymphoma cell leukemia, respectively. The mice in group C developed typical clinical signs of a GVHD after BMT with an average survival time(AST) of (11.50 +/- 3.50) d. The signs of aGVHD were less evident in the group D and E, and their AST (21.70 +/- 5.80 and 25.80 +/- 5.20 days, respectively) were all longer than that in group C (P < 0.05). AST of group E was the longest (P < 0.05). The mice in group B all died of leukemia within 18 days after engraftment of EL4 cells. There was was no significant difference in groups C, D and E in the incidence of leukemia (P > 0.05). (2) Serum IFN-gamma level reached peak value. At + 12 d, then decreased gradually in group C, D, and E, and then reached the nadir at +18 d post-BMT, with the lowest in group E (P < 0.05), and the level was significantly lower in group D than in group C (P < 0.05). After BMT, serum IL-4 level slightly decreased in group C, but gradually elevated in group D and E and reached their peak at +12 d, and even more significantly increased in group E (P < 0.05). There was no statistical significance in the pair wise comparison among three group (P < 0.05). (3) The average proportion of H-2d positive cells in RM was 95%-100% on day 30 post-BMT, with complete donor-type implantation.</p><p><b>CONCLUSION</b>Immature DC can induce immuno tolerance. Immature DC genetically modified to express sTNFR I has been shown to prevent acute GVHD in lethally irradiated mice reconstituted with allogeneic bone marrow grafts while maintaining the GVL response.</p>


Subject(s)
Animals , Bone Marrow Transplantation , Methods , Dendritic Cells , Allergy and Immunology , Female , Graft vs Host Disease , Graft vs Leukemia Effect , Immune Tolerance , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Receptors, Tumor Necrosis Factor, Type I , Genetics , Transplantation, Homologous
13.
Chinese Medical Journal ; (24): 911-917, 2011.
Article in English | WPRIM | ID: wpr-239925

ABSTRACT

<p><b>BACKGROUND</b>Atherosclerotic plaque rupture is the primary mechanism of thrombosis which plays a key role in the onset of acute coronary syndromes. Detection of these plaques prone to rupture (vulnerable plaque) could be clinically significant for prevention of cardiac events. It has been shown that high metabolism cells have a high uptake of fluorine-18 fluorodeoxyglucose ((18)F-FDG). The objective of this study was to investigate the correlation of FDG uptake and the immuno-histochemistry parameters of plaques, and the effect of atorvastatin on vulnerable atherosclerotic plaque in a rabbit model.</p><p><b>METHODS</b>Ten male New Zealand White rabbits were divided into three groups as follows: (1) normal control group (n = 2, C group): the animals were fed a standard diet at 120 g/d and were given water ad labium; (2) atherosclerosis group (n = 4, As group): animals were fed with high fat diet for 5 months after aortic endothelia damage; (3) treatment group (atherosclerosis + atorvastatin, n = 4, Statin group): animals were fed with high fat diet for 5 months and then changed into normal chow plus atorvastatin (2.5 mg·d(-1)·kg(-1)) treatment for another 4 months. Then these four rabbits were imaged with fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) and sacrificed for pathohistologic studies. FDG uptake by the aorta was expressed as target-to-background ratio (TBR). Maximal standardized uptake value (SUV) was measured over the thoracic and abdominal aortas. The aortic smooth muscle cell (SMC) number, CD-14 antibody positive cell (macrophage) number and the ratio of the thickness of fibrous cap to the thickness of lipid core (cap-to-core ratio) in atherosclerotic plaques were analyzed.</p><p><b>RESULTS</b>As group showed significantly higher uptake of FDG than C group (SUVs: 0.746 ± 0.172 vs. 0.286 ± 0.073, P < 0.001). After 4 months of atorvastatin treatment and the modification of diet, SUVs decreased significantly (Statin group: 0.550 ± 0.134, compared to As group, P < 0.001). However, no marked difference was found in TBR, the number of macrophages, the number of SMC and the cap-to-core ratio in the aortic segments between Statin group and As group. The correlation of aortic FDG uptake with SMC assessed by histopathology was negatively significant (r = -0.57, P < 0.001). When aortic FDG uptake was expressed as TBR, it correlated significantly (r = 0.69, P < 0.001) with the macrophage number, and also correlated significantly (r = -0.78, P < 0.001) with the cap-to-core ratio.</p><p><b>CONCLUSION</b>(18)F-FDG PET/CT might serve as a useful non-invasive imaging technique for detection of atherosclerotic plaque and potentially permit monitoring of relative changes in inflammation within the atherosclerotic lesion.</p>


Subject(s)
Animals , Aorta , Diagnostic Imaging , Pathology , Atherosclerosis , Diagnostic Imaging , Fluorodeoxyglucose F18 , Male , Plaque, Atherosclerotic , Diagnostic Imaging , Positron-Emission Tomography , Methods , Rabbits
14.
Article in Chinese | WPRIM | ID: wpr-230318

ABSTRACT

This study was purposed to constructe the three-plasmid system of the lentiviral vector carrying the green fluorescent protein (GFP) gene and to investigate the expression of GFP in T lymphocytes of the mouse. The polypurine tract (PPT) element, ubiquinone promoter (PUB) and GFP were ligated to plasmid pLO134 using subcloning technology to construct plasmid pTK153. Human kidney 293T cells were co-transfected with the three-plasmid system containing packaging plasmid DeltaNRF, plasmid pTK153 and envelope plasmid VSV-G by using calcium phosphate DNA precipation and the expression of GFP was observed under fluorescence microscope after 12 hours. The viral particles were collected after transfection 72 hours, were frozen at -80 degrees C and were used to infect mouse T lymphocytes at multiplicity of infection (m.o.i.) of 3. The expression of GFP in mouse T lymphocytes was observed by fluorescence microscopy and fluorescence-activated cell sorting (FACS). The results showed that the transfection efficacy was 63.04 +/- 7.24% in 293T cells analysed by FACS and the viral titer was (3.09 +/- 0.61) x 10(6) U/ml. The expression of GFP was also evident in mouse T lymphocytes and the transduction efficacy was (37.98 +/- 6.26)%. It is concluded that the three-plasmid system of lentiviral vector containing GFP gene is successfully constructed and the transduction efficacy is high in mouse T lymphocytes.


Subject(s)
Animals , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Lentivirus , Genetics , Mice , Mice, Inbred BALB C , RNA, Viral , T-Lymphocytes , Metabolism , Transduction, Genetic
15.
Chinese Journal of Hematology ; (12): 727-730, 2007.
Article in Chinese | WPRIM | ID: wpr-262958

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effects of Tripterygium hypoglaucum (level) Hutch (THH) on cytokine production in acute graft-versus-host disease (aGVHD)mice and explore the mechanisms.</p><p><b>METHODS</b>2 x 10(7) bone marrow cells mixed with 2 x 10(7) spleen cells from the same C57BL/6 mouse were transplanted into the myeloablative irradiated inbred BALB/c mouse to establish a aGVHD model. The experiments were designed as follows: control group (group A), CsA prophylaxis group (group B), THH prophylaxis group (group C), and combined THH with CsA prophylaxis group (group D). aGVHD was assessed by histologic changes of skin, liver and intestines. Chimerism was detected by H-2b molecular expression on recipient mice bone marrow cells with flow cytometer. Serum concentrations of IFN-gamma, IL-4 and IL-10 were determined by ELISA.</p><p><b>RESULTS</b>The serum concentrations of IFN-gamma in group B, C, D were significantly lower than that in group A, while those of IL-10 was significantly higher than that in group A (P < 0.05). There was no changes in concentration of IL-4 in all the groups (P > 0.05). The median survival time for group A was nine days, while that of group B, C, D each was more than 30 days being significantly longer than that of group A (P < 0.05). The recipient mice of group A displayed significant clinical symptoms of GVHD, and died within 20 days; whereas those of group B, C, D showed only ruffled fur and uplift posture. Histologic changes of liver and intestines in group B and C displayed a few lymphocytes infiltration while the histologic morphology of skin, liver and intestines in the survived mice of group D was normal. Allogeneic chimerism rates of group B, C, D at day 30 after allo-BMT were (99.18 +/- 0.58)%, (97.68 +/- 0.59)%, and (99.15 +/- 0.11)%, respectively.</p><p><b>CONCLUSION</b>THH could regulate the production of cytokines and prevent aGVHD. THH and CsA at low dose combination showed synergic effect in preventing aGVHD.</p>


Subject(s)
Animals , Bone Marrow Transplantation , Disease Models, Animal , Female , Graft vs Host Disease , Blood , Interferon-gamma , Blood , Interleukin-10 , Blood , Interleukin-4 , Blood , Lymphocyte Transfusion , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Phytotherapy , Tripterygium , Chemistry
16.
Article in Chinese | WPRIM | ID: wpr-356497

ABSTRACT

To construct retroviral vector carrying green fluorescent protein (GFP) gene, and determine whether T cells can be infected by retrovirus, the phosphoglycerate kinase (PGK) promoter gene and GFP full-length cDNA were subcloned into the retroviral vector pLXSN. The recombinant vector was transfected into PA317 packaging cells by DNA-calcium phosphate coprecipitation. The G418 resistant clones were selected, then NIH3T3 cells and T cells were infected by the culture supernatant of the retrovirus containing GFP. The results showed that GFP expression in packaging cell line PA317 was detectable under fluorescence microscope, which demonstrated that the GFP retrovirus were able to infect T cells. It is concluded the retrovirus vectors can introduce a foreign gene into T cells efficiently and can be used as important tools to deliver gene into T cells.


Subject(s)
Animals , Cell Line , Cells, Cultured , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Mice , Microscopy, Fluorescence , NIH 3T3 Cells , Recombinant Fusion Proteins , Genetics , Metabolism , Retroviridae , Genetics , T-Lymphocytes , Cell Biology , Metabolism , Transfection , Methods
17.
Chinese Medical Journal ; (24): 474-479, 2005.
Article in English | WPRIM | ID: wpr-250904

ABSTRACT

<p><b>BACKGROUND</b>Nonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).</p><p><b>METHODS</b>A total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Co gamma-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3 x 10(7) bone marrow cells and 1 x 10(7) spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells (2 x 10(7)) on day 14 or 21, or donor spleen cells (5 x 10(7)) pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.</p><p><b>RESULTS</b>The median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P < 0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.</p><p><b>CONCLUSION</b>Donor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.</p>


Subject(s)
Animals , Bone Marrow Transplantation , Allergy and Immunology , Cyclosporine , Pharmacology , Female , Graft vs Host Disease , Graft vs Leukemia Effect , Allergy and Immunology , Hydrocortisone , Pharmacology , Lymphocyte Activation , Lymphocyte Transfusion , Male , Mice , Mice, Inbred BALB C , Transplantation Chimera , Transplantation, Homologous
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