Expression of MiR-155 in Tissue of Patients with Diffuse Large B-Cell Lymphoma and Its Effect on Cell Biological Characteristics / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of miR-155 in patients with diffuse large B-cell lymphoma (DLBCL), and to explore the effect of transfection of miR-155 inhibitor on the biological characteristics of DLBCL cells.@*METHODS@#A total of 76 patients with DLBCL treated in our hospital were selected from April 2013 to December 2017. In the same time, 40 cases of lymph node reactive hyperplasia (LNRH) were selected as control group. DB cells were cultured and divided into miR-155 inhibitor, negative control and blank groups. The expressions of miR-155 in DLBCL, negative and blank control groups were detected by using real-time PCR, the cell proliferation was detected by MTT assay, the apoptosis was detected by flow cytometry, and the cell migration and invasion were detected by Transwell assay.@*RESULTS@#The relative expression level of miR-155 in tissues of DLBCL patients was significantly higher than that in tissne of controls (1.93±0.16 vs 1.01±0.09) (t=33.991, P=0.000). The expression level of miR-155 increased (P＜0.05) in DLBCL patients with LDH level abnormarity, BCL-2, MUM1, Ki-67≥50%, non-GC type, Ann Arbor stage III-IV, extranodal lesion number≥2 and IPI score 3-5. The relative expression level of miR-155 in the miR-155 inhibitor group was lower than that in the negative control group and the blank group (P＜0.05). The absorbance (A) values at 24, 48, 72 and 96 h of culture in the miR-155 inhibitor group were lower than those in the negative control group and the blank group (P＜0.05), while the apoptotic rate was higher than that in the negative control group and the blank group (P＜0.05). Both the migrating cells and invading cell number in the miR-155 inhibitor group were lower than those in the negative control group and the blank group (P＜0.05).@*CONCLUSION@#The miR-155 highly expresses in DLBCL tissue, which relates with tumor malignancy and invasion progression. The specific inhibition of miR-155 expression in DB cells can reduce cell proliferation, accelerate cell apoptosis, and inhibit cell migration and invasion.

Kinetic study of various cytokine mRNA expressions in rhesus treated with haploidentical peripheral blood stem cell transplantation / 中国实验血液学杂志

This study was aimed to analyze the mRNA expression of cytokines (TGF-beta, IL-2, IL-6, IL-10, IFN-gamma, TNF-alpha, FAS-L) in five rhesus treated with haploidentical peripheral blood stem cell transplantation after nonmyeloablative preparative regimens and to explore the role of these cytokines in the development and pathology of acute graft-versus-host-disease (aGVHD). Five rhesus monkeys received nonmyeloablative haploidentical peripheral blood stem cells transplantation. Semi-quantitative reversed transcription polymerase chain reaction (RT-PCR) was used to analyze the kinetics of cytokine mRNA expression in the transplantation and aGVHD. The results showed that five rhesus monkeys acquired hematopoietic reconstitution successfully. The graft was rejected in one monkey which survived without disease, the other four achieved mixed chimerism and full donor chimerism. Chimerism of low centigrade in one monkey achieved high centigrade at 35 days after donor stem cell infusion. Intestinal aGVHD grade III developed in one monkey. Cytokines of Th1 and Th2 changed after transplantation. In period of aGVHD, expression of TGF-beta decreased but all others increased in various levels. When donor chimerism decreased, the cytokines decreased accordingly. It is concluded that the decrease of TGF-beta mRNA may be an indicator to predict aGVHD, and can be used as a differential diagnostic indicator for intestinal GVHD.

Establishment of rhesus model for haploidentical hematopoietic stem cell transplantation with nonmyeloablative conditioning / 中国实验血液学杂志

To study if rhesus haploidentical hematopoietic stem cell transplantation model can be established by non-myeloablative conditioning, parent monkeys were used as donors, offspring monkeys were used as recipients. The recipient monkeys received a nonmyeloablative conditioning consisting of fludarabine, cyclophosphamide, total body irradiation and rabbit anti-human thymocyte globulin. Cyclosporine, mycophenolate mofetil and anti CD25 antibody were used for GVHD prevention. Donor mobilized peripheral blood stem cells were transplantated on day 0. Hematopoietic recovery, chimerism level, GVHD were assessed regularly. The results indicated that hematopoietic recoveries in all 4 cases were observed within 8 days after transplantation. Donor hematopoietic chimerism could be induced in all cases, chimerism analysis showed full donor chimerism (FDC) in case 3 and 4, and II to III grade GVHD developed on day 12 and 14. In case 1, only low level donor chimerism was detected on day 7, and transplantation rejection happened eventually. Unfortunately, kidney failure happened in case 2 after conditioning and died several days later, chimerism analysis showed 50% donor rate on day 7. It is concluded that the rhesus transplantation model was successfully established by nonmyeloablative conditioning for striding over the MHC barrier. This rhesus monkey model would provide a basis for future research.

Establishment and application of a method for assessing hemopoietic chimerism in rhesus after allogeneic stem cell transplantation / 中国实验血液学杂志

Monitoring engraftment of donor cells after allogeneic transplantation is the key of assessing successful establishment of animal transplantation model. The purpose of this study was to establish a method for analysis of chimerism in rhesus transplantation model. Y-specific sequence in rhesus was amplified by the polymerase chain reaction (PCR), method for analysis of chimerism in rhesus after sex-mismatched transplantation was established; the feasibility and sensitivity of the approach were tested by using serial DNA mixtures of sex-mismatched individuals; the accuracy of results was confirmed by chromosome karyotype analysis simultaneously; Chimerisms of one rhesus received allogeneic stem cell transplantation and the other received mesenchymal stem cells (MSC) transfusion were detected by this method. The results showed that a 176 bp long sequence of PCR product was gained in male rhesus, while no product was gained in female rhesus. The sensitivity of this method was up to 0.05% (male/female DNA ratio). Male donor chimerism were found on day 7 and 14 after allogeneic stem cell transplantation by Y-specific sequence and chromosome karyotype analysis. Otherwise, male donor chimerism was found in peripheral blood at 1 hour and in bone marrow on day 30 after MSC transfusion by this method, but no male donor chimerism was found after MSC transfusion using chromosome karyotype analysis. In conclusion, this rapid, sensitive approach can used to assess chimerism in experiments of rhesus alloorgan transplantation and cell transfusion.

Establishment of a rhesus haploidentical hematopoietic stem cell and mesenchymal stem cell transplantation model by nonmyeloablative conditioning / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish rhesus haploidentical hematopoietic stem cell transplantation model by nonmyeloablative conditioning, and examine the effects of mesenchymal stem cells (MSC) in haploidentical transplantation.</p><p><b>METHODS</b>The recipient haploidentical rhesus monkeys were conditioned with a nonmyeloablative regimen consisted of fludarabine, cyclophosphamide, 200 cGy total body irradiation, and rabbit anti-human thymocyte globulin. Cyclosporine A, mycophenolate mofetil and anti CD25 antibody were used for graft versus host disease (GVHD) prophylaxis. Rhesus monkeys in one group were given hematopoietic stem cell (HSC) only, while in the other group HSC combined with MSC. The differences in hematopoiesis recovery, chimerism level, and GVHD between the two groups were evaluated.</p><p><b>RESULTS</b>Stable chimerism could be achieved in recipient monkeys. Hematopoiesis recovery was mainly related with chimerism level. MSC seemed capable of facilitating HSC engraftment, as there were more mixed chimerism and less GVHD occurrence in the HSC combined with MSC recipient group.</p><p><b>CONCLUSION</b>A rhesus haploidentical hematopoietic stem cell transplantation model is successfully established by nonmyeloablative conditioning. MSC was of great benefit to haploidentical transplantation.</p>