Allogeneic Hematopoietic Stem Cell Transplantation for Paroxysmal Nocturnal Hemoglobinuria / 中国实验血液学杂志

OBJECTIVE@#To analyze the clinical efficacy and safety of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH), and preliminarily explore the role of an improved post-transplantation cyclophosphamide (PTCy) based conditioning regimen in PNH patients receiving transplantation.@*METHODS@#Clinical related data of PNH sufferers receiving allo-HSCT in Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology were collected, and hematopoietic reconstitution, chimerism, PNH cloning, graft-versus-host disease (GVHD), infection, and survival were analyzed.@*RESULTS@#Totally five PNH patients receiving allo-HSCT were enrolled, including 1 case with classic PNH, 3 cases with aplastic anemia-PNH syndrome, 1 case with myelodysplastic syndrome, three of them (case 1-3) received the improved PTCy based conditioning regimen before HSCT. All sufferers engrafted successfully within 28 days, the median time of neutrophil and platelet engraftment was 11 days and 12 days, respectively, no patient occurred acute or chronic GVHD, after a median follow-up of 16 months, all recipients survived and completely eliminated PNH cloning.@*CONCLUSION@#Allo-HSCT can completely clear PNH cloning and restore hematopoietic function with controllable complications, and the improved PTCy based conditioning regimen is proved to be effective in PNH transplantation.

Inhibitory Effect of Serum Containing Fuzheng Jiedu Decoction on the Leukemia K562/A02 Multi-drug Resistance Cells and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect of serum containing Fuzheng Jiedu decoction on leukemia multi-drug-resistance K562/A02 cells and its possible mechanism.</p><p><b>METHODS</b>The MTT method was used to detect the inhibitory rate of K562/AO2 cells treated with serum containing Fuzheng Jiedu decoction; the flow cytometry was used to detect the inhibitory effect of serum containing medicin on growth of K562/AO2 cells and P-gp expression; the Q-PCR was used to assay the BCL-2 mRNA expression; the Western blot was used to detect the BCL-2 protein expression.</p><p><b>RESULTS</b>MTT cytotoxic test showed serum containing Fuzheng Jiedu decoction could inhibit K562/A02 cell growth, and the inhibitory rate increased with the increase of drug concentration; the flow cytometry showed that the serum containing Fuzheng Jiedu decoction could promote K562/A02 cell apoptosis in a concentration-dependent manner. qPCR and Western blot showed that serum containing Fuzheng Jiedu decoction could down-regulate the protein expression of BCL-2. Fuzheng Jiedu decoction could reduce the protein expression of P-gp on the K562/A02 cell membrane.</p><p><b>CONCLUSION</b>serum containing Fuzheng Jiedu decoction can promote K562/A02 cell apoptosis, its mechanism of inducing apoptosis may be related with the inhibition of BCL-2 and P-gp protein expression.</p>

Gene mutation and myelodysplastic syndromes with ring sideroblast excess / 中国实验血液学杂志

Myelodysplastic syndromes (MDS) are heterogeneous clonal hematopoietic stem cell disorders with different mechanisms and diverse prognosis. The excess of ring sideroblasts (RS) is an important presentation MDS, but the mechanisms of RS appearance are obscure and the treatment of MDS-RS is intractable. Splicing factors play a very important role in the maturation process of eucaryon mRNA, recent studies indicate that there is a significant causal relationship between splicing factor 3B subunit 1 (SF3B1) mutation and the presence of ring sideroblasts. Lucubrating the downstream molecular of the mutated SF3B1 can facilitate exploring the mechanisms and new therapeutic strategies of MDS-RS.

Therapeutic effect of lithium chloride combined with cyclosporine A on mouse model with aplastic anemia / 中国实验血液学杂志

Wnt signaling has been shown to inhibit adipogenic differentiation while inducing the osteogenic pathway of bone marrow stromal cells (BMSC). Patients with aplastic anemia (AA) often show excess fat accumulation in the bone marrow, possibly due to overactivation of the adipogenic pathway. Therefore, an activator of the Wnt signaling may alleviate the symptoms by enhancing the inhibition on the differentiation of BMSC towards adipocytes. To judge this hypothesis, the therapeutic effects of Wnt signaling activator lithium chloride (LiCl) combined with the currently used immunosuppressor cyclosporine A (CsA) on mice with AA in vivo was investigated. Mouse model with AA was established and the disease was confirmed by increased fat cell counts and decreased hematopoietic cell counts in the bone marrow of these animals. These mice treated with CsA 50 mg/(kg·d) alone or together with LiCl 20 mg/(kg·d), once daily for 5 d, then at day 14, 21 and 28 after establishment of mouse model with AA, the treatment effects were observed, including peripheral blood cells, bone marrow mononuclear cells (BMMNC) count and bone marrow biopsy examination in each group. The results showed that compared with the AA group, Hb content, WBC and BMMNC counts of CsA group and the combination group significantly increased. HE staining of bone marrow biopsy sample showed that the fat cells were significantly reduced in the bone marrow cavity (P < 0.05). Compared with the CsA group, Hb content, WBC and BMMNC counts of the combination group significantly increased (P < 0.05); HE staining of bone marrow biopsy sample showed that fat cells were reduced, the hematopoiesis of bone marrow was close to the normal. It is concluded that Wnt signal activator (LiCl) combined with CsA displayed a better treatment effect on AA in mouse models than the effect of using CsA only. They can promote the hematopoietic function of bone marrow, which may correlate with inhibiting differentiation of bone marrow mesenchymal stem cells into the fat cells by Wnt signaling.

Analysis of neutralized antibody level of the poliomyelitis in healthy population in Fujian province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the neutralized antibody level of the poliomyelitis among healthy people and provide scientific evidence for the immunization strategy since routine and intensified immunization with oral polio vaccine (OPV) in Fujian province.</p><p><b>METHODS</b>The poliomyelitis antibody level of healthy people were detected by neutralization test of the micro cells.</p><p><b>RESULTS</b>The neutralizing antibody positive rates were 99.0%, 99.3%, 97.5% and GMTs were 1:79.1, 1:31.2, 1:24.7 for polio I, II, III respectively in 400 serum specimens from 1-59 years old. GMTs present a trend of decreasing as age's increasing.</p><p><b>CONCLUSION</b>A protective barrier had been built against poliomyelitis in healthy people in Fujian province through routine and intensified immunization with OPV.</p>

Relation of notch pathway to senescence of murine bone marrow stromal cells / 中国实验血液学杂志

This study was purposed to investigate the relation of the Notch signaling pathway to senescence of murine bone marrow stromal cells in vitro. Intracellular domain of Notch 1 (ICN) was transfected into cultured murine bone marrow stromal cells by lipofectamine transfection. After transfection for three days the proliferation of transfected cells was measured by MTT, cell cycle distribution was analyzed by flow cytometry. The percentage of senescence associated beta-galactosidase (SA-beta-Gal) positive cells were measured by cytochemical method, and the expression rates of P53 and p21Cip1/Waf1 at gene and protein levels were analyzed by RT-PCR and Western blot respectively. The results showed that after transfection for 3 days the proliferation of murine bone marrow stromal cells was inhibited with induction of G1 arrest, the percentage of SA-beta-gal positive cells increased and the p53 and p21Cip1/Waf1 mRNA and protein expression levels were upregulated. It is concluded that the activated Notch signaling can induce premature senescence of bone marrow stromal cells through the p53-p21Cip1/Waf1 pathway.

Biological characteristics of hematopoietic progenitor cells at different stages of hematopoietic development / 生理学报

The aim of the present paper is to better understand the mechanism of hematopoietic development through studying the biological characteristics of hematopoietic progenitor cells at different stages of development. Firstly, the c-kit expression levels of the mononuclear cells from murine embryonic aorta-gonad-mesonephros (AGM) region at embryonic day (E)10.5 and E11.5, fetal liver (FL) at E12.5, E14.5, E16.5, E18 and bone marrow (BM) were assayed with fluorescence activated cell sorting (FACS). Secondly, hematopoietic progenitor cells derived from AGM at E10.5, FL at E14.5 and BM were isolated by using c-kit microbeads. Isolated c-kit(+) population cells from AGM, FL and BM were then co-cultured with E14.5 FL-derived stromal cells in transwell co-culture system in vitro. After 3, 7, 10 days of co-culture, numerous floating cells were generated. The floating cells generated in transwell inserts were collected for FACS cell count, migration activity detection and colony forming unit (CFU) formation assay. The results showed that the c-kit was highly expressed in E10.5 AGM, with the percentage of c-kit(+) cells declining during AGM development. c-kit expression was highly expressed again in E12.5 FL, declining along with the progressive development of the FL region. Co-cultured with FL-derived stromal cells, E10.5 AGM-derived c-kit(+) cells produced the highest number of hematopoietic cells, while BM-derived c-kit(+) cells produced the lowest number of hematopoietic cells. Compared with E10.5 AGM-derived c-kit(+) cells, E14.5 FL- and BM- derived c-kit(+) cells inclined to differentiate after 7 to 10 days of culture in vitro. E10.5 AGM and E14.5 FL-derived c-kit(+) cells exhibited a higher migration activity than BM-derived c-kit(+) cells. Moreover, E10.5 AGM-derived c-kit(+) cells showed a higher ability to form mixed colony-forming unit (CFU-Mix) colony. In conclusion, compared with FL- and BM-derived c-kit(+) cells, E10.5 AGM-derived c-kit(+) hematopoietic progenitor cells exhibit better proliferation, migration potential, and have a higher ability to maintain the undifferentiation state in vitro, providing an insight into their clinical manipulation.

Establishment of a bone marrow failure mouse model by immune injury / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a mouse model for the study of pathophysiologic mechanism and treatment of bone marrow failure (BMF).</p><p><b>METHODS</b>Balb/c mice (recipient) were irradiated 5.0 Gy by gamma rays of (60)Co, and then infused 5 x 10(6) lymph node (LN) cells from DBA/2 mice (donor) in 4 hours. Pancytopenia was monitored by cell counting, bone marrow damage was assessed by histological staining and mononuclear cell counting. Serum IFN-gamma concentration was measured by ELISA. The proportion of Treg in spleen was detected by flow cytometry.</p><p><b>RESULTS</b>Irradiation and infusion of LN cells led to rapid development of severe pancytopenia and BM hypoplasia, which reached the most severity at d14. The pancytopenia remained at d28 and displayed no signs of recovery. The bone marrow was full of adipose cells with scarcity of hematopoietic cells at d14 and persisted at least for 28 days, being similar to the feature of aplastic anemia. Serum IFN-gamma concentration was 6.3 fold increased \[(170.0 +/- 17.0) vs (27.7 +/- 7.1) pg/ml\] at d6. Tregs were decreased after infusion, and then increased \[(3.38 +/- 0.52)%\] and recovered to normal \[(4.04 +/- 0.44)%\] at d21. The expression level of the specific transcription factor Foxp3 was similar to normal.</p><p><b>CONCLUSION</b>The MHC antigen of Balb/c mice is identical to that of DBA/2 mice, but their minor antigen differs. 5.0 Gy irradiation and then 5 x 10(6) lymphocyte infusion can induce BMF similar to the features of aplastic anemia.</p>

Study on Wnt and Notch signalling involves in regulation of hematopoietic microenvironment / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of Wnt and Notch pathway involved modulating time and spatial restricted hematopoiesis.</p><p><b>METHODS</b>Murine hematopoietic stem and progenitor cells (HSPCs) were isolated from bone marrow (BM) by using c-kit microbeads. E10.5 aorta-gonad-mesonephros (AGM), E12.5, E14.5, E16.5 fetal liver (FL) and adult BM derived stromal cells (StroCs) were isolated and co-cultured with c-kit(+)HSPCs. The floating cells in co-culture system were sorted and counted by FACS. Gene expressions of Wnt and Notch pathway were detected by quantitative PCR and protein expressions by immunostaining.</p><p><b>RESULTS</b>Co-culturing HSPCs with AGM and FL-derived StroCs resulted in an expansion of c-kit(+)population from 0.4 x 10(5)/well to (19.2 +/- 3.2) x 10(5)/well and (26.8 +/- 5.4) x 10(5)/well, respectively, being greater than that with BM-derived StroCs (P < 0.05). The percentage of c-kit(+)cells detected in AGM- and BM- derived StroCs culture system was (75.2 +/- 7.1)%, (74.1 +/- 6.2)% respectively, being higher than FL- derived StroCs culture system (63.4 +/- 5.3)% (P < 0.05). Wnt and Notch pathway genes expression varied at different phases of hematopoiesis. Wnt was highly expressed in AGM and FL derived StroCs, and, Notch did in AGM and BM derived StroCs.</p><p><b>CONCLUSION</b>Wnt and Notch pathway are important modulators in regulating time and spatial restricted hematopoiesis.</p>

Clinical study on needling method for regulating mental activities and soothing liver for treatment of melancholia / 中国针灸

<p><b>OBJECTIVE</b>To observe clinical therapeutic effect of needling method for regulating mental activities and soothing liver on melancholia.</p><p><b>METHODS</b>One hundred and ten cases of depression were selected according to the diagnostic criteria of depression in mood disturbance (emotional and mental disturbance) and randomly divided into a regulating mental activities and soothing liver needling group (n=50), a traditional acupuncture group (n=30) and a prozac treatment group (n=30). Their therapeutic effects were evaluated by Hamilton depression scale, Self-rating depression scale (SDS), psychiatric self-rating scale (SCL-90).</p><p><b>RESULTS</b>In the regulating mental activities and soothing liver needling group, 34 cases were cured, 10 cases were markedly effective, 4 cases were effective and 1 cases were ineffective, with the cured rate of 69.4% and the total effective rate of 98.0%, which were significantly better than those in the traditional acupuncture group (the cured rate was 48.3% and the total effective rate was 89.7%), and was not significantly different with the prozac treatment group (the cured rate was 64.0% and the total effective rate was 96.0%).</p><p><b>CONCLUSION</b>The needling method for regulating mental activities and soothing liver has a better therapeutic effect on depression.</p>