Effect of consolidation before allogeneic hematopoietic stem cell transplantation for non-favorable acute myeloid leukemia patients with first complete remisson and negative minimal residual disease / 中华血液学杂志

Objective: To probe the prognostic value of consolidation chemotherapy in non-favorable acute myeloid leukemia (AML) patients who were candidates for allogeneic hematopoietic stem cell transplantation (allo-HSCT) with first complete remission (CR(1)) and negative minimal residual disease (MRD(-)) . Methods: A retrospective analysis was conducted on 155 patients with non-favorable AML who received allo-HSCT in CR(1)/MRD(-) from January 2010 to March 2019. The survival data were compared between patients who received and those not received pre-transplant consolidation chemotherapy. Results: A total of 102 patients received pre-transplant consolidation chemotherapy (consolidation group) , and 53 cases directly proceeded to allo-HSCT when CR(1)/MRD(-) was achieved (nonconsolidation group) . The median ages were 39 (18-56) years old and 38 (19-67) years old, respectively. Five-year post-transplant overall survival [ (59.3±7.5) % vs (62.2±6.9) %, P=0.919] and relapse-free survival [ (53.0±8.9) % vs (61.6±7.0) %, P=0.936] were not significantly different between the two groups (consolidation vs nonconsolidation) . There was a weak relationship between consolidation therapy and cumulative incidence of relapse [consolidation: (21.9±5.4) % vs nonconsolidation: (18.3±6.0) %, P=0.942], as well as non-relapse mortality [consolidation: (22.4±4.3) % vs nonconsolidation: (28.4±6.5) %,P=0.464]. Multivariate analysis indicated that pre-transplant consolidation and the consolidation courses (< 2 vs ≥2 courses) did not have an impact on allo-HSCT outcomes. Conclusion: Allo-HSCT for candidate patients without further consolidation when CR(1)/MRD(-) was attained was feasible.

Effect of SET7/9-mediated endoplasmic reticulum stress on arsenic-induced hepatocyte apoptosis / 中国病理生理杂志

AIM:To investigate the effect of SET7/9 (SET domain containing 7/9) -mediated endoplasmic reticulum stress (ERS) on protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway, and to explore the mechanisms of arsenic-induced hepatocyte apoptosis.METHODS:Human liver LO2 cells were divided into control group, arsenic poisoning model group, negative transfection group and SET7/9 siRNA transfection group.The apoptosis of the LO2 cells in each group was analyzed by flow cytometry.The protein levels of SET7/9, glucose-regulated protein 78 (GRP78) , PERK and p-PERK in the LO2 cells of each group were observed by Western blot.RESULTS:Inhibition of SET7/9 expression reduced the apoptotic rate of arsenic-induced LO2 cells.Arsenic exposure increased the expression of SET7/9 in the LO2 cells.Arsenic exposure increased the protein levels of GRP78 and p-PERK in the LO2 cells, but decreased the protein levels of GRP78 and p-PERK after transfection with SET7/9 siRNA (P<0.05).CONCLUSION:Arsenic exposure induces hepatocyte apoptosis by increasing SET7/9 to activate ERS by PERK signaling pathway.

Prognostic value of donor chimerism at +90 days after allogeneic hematopoietic stem cell transplantation in young patients with intermediate-risk acute myeloid leukemia / 中华血液学杂志

Objective: To investigate the relationship between donor chimerism and relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: The clinical data of 105 patients with acute myeloid leukemia (AML) who underwent allo-HSCT and recurrence-free survival>90 days from January 2010 to January 2019 were retrospectively analyzed. The bone marrow samples were collected at 15, 30, 60, 90, 180, 270, 360 days after transplantation. Donor chimerism was detected by single nucleotide polymorphism (SNP) -PCR. Results: Of the 105 patients, 43 cases were male and 62 cases were female, with a median age of 38 (16-60) years. Till April 2019, the median follow-up was 843 (94-3 261) days. Ninety days after transplantation, 18 cases relapsed, 33 cases died, and 72 cases survived. The 3-year overall survival (OS) rate was (66.8±5.1) %, and the recurrence-free survival (RFS) rate was (65.1±5.0) %. Pre-transplant disease status, pre-transplant minimal residual disease (MRD) , and 90 day post-transplantation chimerism were independent risk factors related to RFS. The risk of recurrence was significantly increased in patients with a donor chimerism rate ≤97.24% at 90 days after transplantation[HR=6.921 (95%CI 2.669-17.950) , P<0.001], which was considered as a sign of early relapse. Conclusion: SNP-PCR is an applicable method for detecting donor chimerism in patients after allo-HSCT. Chimerism rate equal or less than 97.24% at 90 days after transplantation predicts a higher risk of relapse.

Clinical and prognostic values of TP53 mutation in patients with acute myeloid leukemia / 中华血液学杂志

Objective: To explore the clinical and prognostic values of TP53 gene mutation in patients with acute myeloid leukemia (AML) . Methods: A retrospective analysis of 265 newly diagnosed AML patients with next-generation sequencing (NGS) data in the Hematology Department of Changhai Hospital from January 2010 to January 2019 was performed. Mutation analysis was carried out by targeted sequencing technology including 200 hematological malignancy related genes. The association of TP53 mutation with clinical features was analyzed. Results: Alterations in TP53 were found in 20 (7.5%) patients, including 17 case (6.4%) of missense mutations, 2 cases (0.7%) of frame-shift deletion mutations and 1 case (0.4%) of splicing sites mutation. A total of 23 kinds of TP53 mutations were detected, most of them (16, 69.6%) were located in the DNA binding domain of exon 5-8, 4 in the DNA binding domain of exon 3-4, 2 in exon 10 and 1 in splice site, respectively. The median age of patients with TP53 alterations was higher than those without [52 (26-72) years old vs 45 (14-75) years old, P= 0.008]. The frequency of complex karyotypes was higher in patients with TP53 alterations than those without [45.0% (9/20) vs 6.1% (15/245) , P<0.001]. Median overall survival (OS) of patients with TP53 alterations was shorter than those without[14.1 (95%CI 6.78-21.42) months vs 31.4 (95%CI 13.20-49.59) months, P=0.029]. The OS of patients treated with "Decitabine + CAG" was superior than that of patients treated with "3 + 7" regimen [30.0 (95%CI 27.35-38.84) months vs 12.5 (95%CI 5.80-19.19) months, P=0.018]. Multivariate analysis indicated that TP53, DNMT3A and USH2A alterations, WBC ≥ 12.45×10(9)/L had negative impacts on OS. Conclusion: The frequency of TP53 mutation was 7.5% in our cohort. Most mutations were located in the DNA binding domain. TP53 alterations were strongly associated with older age, complex karyotype and shorter OS. Decitabine-based induction chemotherapy and hematopoietic stem cell transplantation may improve OS, more cases and/or multicenter randomized studies are needed for further confirmation.

Rosai-Dorfman disease: a clinicopathologic analysis and whole exome sequencing in 23 cases / 中华血液学杂志

Objective: To evaluate the clinicopathologic features of Rosai-Dorfman disease (RDD) , and elucidate the potential pathogenesis by whole exome sequencing (WES) . Methods: Clinico-pathological data of 23 RDD patients diagnosed between 2010 and 2018 in Changhai hospital were reviewed, and 9 paraffin-embedded specimens were performed for WES. Results: The median age of 23 RDD patients was 47 (10-79) years. Of them, 19 cases had extranodal lesions, 3 had nodal lesions, and 1 had nodal and extranodal lesions coincidently. All patients received surgery for lesion resection. Histiocytosis in lymph node sinuses or in extranodal tissues accompanied by lymphocyte phagocytosis are typical pathological features of RDD. Immunohistochemical staining shows histocytes are positive for S100, CD68 and CDl63, and negative for CD1a. mTOR, KMT2D and NOTCH1 mutations were detected with WES in these cases. Conclusion: Mutations in mTOR, KMT2D and NOTCH1 genes may be involved in the pathogenesis of RDD, and their clinical significance needs to be further studied.

Efficacy of Hyper-CVAD/MA and CHALL-01 regimens in the treatment of Philadelphia chromosome-positive adult acute lymphoblastic leukemia patients under 60 years old / 中华血液学杂志

Objective: To compare the difference of efficacy between traditional Hyper-CVAD/MA regimen and the adolescents inspired chemotherapy regimen, CH ALL-01, in treatment of adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL) . Methods: In this study we retrospectively analyzed 158 Ph(+) ALL patients receiving Hyper-CVAD/MA regimen (n=63) or CHALL-01 regimen (n=95) in our center and Changzheng hospital from January 2007 to December 2017, excluding patients with chronic myeloid leukemia in blast crisis. Tyrosine kinase inhibitor (TKI) was administered during induction and consolidation chemotherapy. Patients who underwent hematopoietic stem cell transplantation received TKI as maintenance therapy. Results: Of them, 91.1% (144/158) patients achieved complete remission (CR) after 1-2 courses of induction. CR rate was 90.5% (57/63) for patients in Hyper-CVAD/MA group and 91.6% (87/95) for patients in CHALL-01 group. There was no difference in CR rates between the two groups (χ(2)=0.057, P=0.811) . The last follow-up was June 2018. A cohort of 134 CR patients could be used for further analysis, among them, 53 patients received Hyper-CVAD/MA regimen and other 81 patients received CHALL-01 regimen. The molecular remission rates were significantly higher in CHALL-01 group (complete molecular response: 44.4%vs 22.6%; major molecular response: 9.9% vs 18.9%) (χ(2)=7.216, P=0.027) . For the patients in Hyper-CVAD/MA group, the 4-year overall survival (OS) was 44.81% (95%CI: 30.80%-57.86%) and the 4-year disease free survival (DFS) was 37.95% (95%CI: 24.87%-50.93%) . For patients received CHALL-01 regimen, the 4-year OS was 55.63% (95%CI: 39.07%-69.36%) (P=0.037) and 4 year DFS was 49.06% (95%CI: 34.24%-62.29%) (P=0.015) , while there was no significant difference in 4 year cumulative incidence of relapse (CIR) (P=0.328) or cumulative incidence of nonrelapse mortality (CI-NRM) (P=0.138) . The rate of pulmonary infection was lower in patients received CHALL-01 regimen compared with patients received Hyper-CVAD regimen (43.4% vs 67.9%, χ(2)=7.908, P=0.005) . Conclusions: Outcome with CHALL-01 regimen appeared better than that with the Hyper-CVAD/MA regimen in Ph(+) ALL, which has lower incidence of pulmonary infection, higher molecular remission rate and better OS and DFS.

Precision therapy of acute myeloid leukemia: A new focus / 第二军医大学学报

Acute leukemia is a malignant tumor with the highest morbidity and mortality in patients younger than 35 years old. Three-year overall survival of middle-risk patients receiving conventional chemotherapy is only 30%-50%, although the stratified chemotherapy based on cell and molecular genetics has improved the overall survival in recent years. To further optimize the treatment, we used flow cytometry in combination with fluorescent in situ hybridization to detect the competing of leukemia stem cells with hemopoietic stem cell, which could diagnose the relapse of patients 2-3 months ahead of time, thus allowing early intervention and improving the survival rate. In allogeneic hematopoietic stem cell transplantation, we have designed a novel conditioning regimen, which balanced the graft-versus-host disease and graft-versus-leukemia effect and reduced transplant-related mortality. This is a new focus on acute leukemia treatment and a further extension of precision therapy in leukemia.

Research Progress on Thrombocytopenia after Allogeneic Hematopoietic Stem Cell Transplantation -Review / 中国实验血液学杂志

Thrombocytopenia is a major complication following allogeneic stem cell transplantation(allo-HSCT). Overall survival(OS) and disease-free survival(DFS) of patients with thrombocytopenia were lower than those without thrombocytopenia. Lower platelet counts before conditioning, graft-versus-host disease(GVHD) and cytomegalovirus(CMV) infection are adverse factors for the patiens with thrombocytopenia. Bone marrow microenvironment may be involved in the pathogenesis. Thrombopoietin(TPO) and mesenchymal stem cells can improve the platelet counts. In this review the definition, prognosis, pathogenesis and potential therapy for thrombocytopenia after allo-HSCT are summarized.

Clinical and laboratory characteristics of hematological diseases associated with eosinophilia / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the clinical and laboratory characteristics of hematological diseases associated with eosinophilia.</p><p><b>METHODS</b>Karyotype analysis was performed by direct method and/or short-time culture of bone marrow cells for R-banding. Fluorescence in situ hybridization (FISH) was performed using PDGFRα, PDGFRβ and FGFR1 break-apart probes.</p><p><b>RESULTS</b>The clinical and hematological findings of 44 patients were diagnosed as hematological diseases associated with eosinophilia. Abnormal karyotypes were detected in 6 cases (13.64%) with karyotyping. The efficiency of the detection of abnormal clone was markedly increased to 29.55% (13/44) with FISH techniques, including 7 cases with FIP1L1-PDGFRα (F/P, 15.91%), 3(6.82%) PDGFRα rearrangement, 2 (4.55%) aberrant PDGFRβ gene and 1(2.27%) FGFR1 rearrangement. Patients being PDGFRα, PDGFRβ or FGFR1 positive (13 cases) or negative (31 cases) showed predominant difference in clinical and laboratory features. The incidence of gut involvement, the absolute count of eosinophils in peripheral blood and the percentage of immature eosinophils in bone marrow were significantly increased in positive patients (P < 0.05).</p><p><b>CONCLUSIONS</b>The hematological diseases associated with eosinophilia are characterized by unique clinical and laboratory features. Karyotyping should be a routine approach to detect the abnormal clone in these diseases. Screening for PDGFRα, PDGFRβ and FGFR1 gene with FISH can provide more genetic information.</p>

Discussion on the teaching strategies of pharmaceutical vocabulary / 中华医学教育探索杂志

Professional terms are the key points in the teaching of pharmaceutical English.By taking the strategies of comparing amphibious words,analyzing morphemes,emphasizing on phonetics,providing study material in real context and strengthening review,the teaching of pharmaceutical vocabulary can be efficiently performed and interest in learning stimulated.

Comparison of bupivacaine, ropivacaine and levobupivacaine with sufentanil for patient-controlled epidural analgesia during labor: a randomized clinical trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Ropivacaine and levobupivacaine have been introduced into obstetric analgesic practice with the proposed advantages of causing less motor block and toxicity compared with bupivacaine. However, it is still controversial whether both anesthetics are associated with any clinical benefit relative to bupivacaine for labor analgesia. This study aimed to compare the analgesic efficacy, motor block and side effects of bupivacaine, ropivacaine and levobupivacaine at lower concentrations for patient-controlled epidural labor analgesia.</p><p><b>METHODS</b>Four hundred and fifty nulliparous parturients were enrolled in this randomized clinical trial. A concentration of 0.05%, 0.075%, 0.1%, 0.125% or 0.15% of either bupivacaine (Group B), ropivacaine (Group R) or levobupivacaine (Group L) with sufentanil 0.5 microg/ml was epidurally administered by patient-controlled analgesia mode. Effective analgesia was defined as a visual analogue scale score was <or=30 mm. The relative median potency for each local anesthetic was calculated using a probit regression model. Parturients demographics, sensory and motor blockade, obstetric data, maternal side effects, hourly volumes of local anesthetic used, and others were also noted.</p><p><b>RESULTS</b>There were no significant differences among groups in the numbers of effective analgesia, pain scores, hourly local anesthetic amount used, sensory and motor blockade, labor duration and mode of delivery, side effects and maternal satisfaction (P>0.05). The relative median potency was bupivacaine/ropivacaine: 0.828 (0.602-1.091), bupivacaine/levobupivacaine: 0.845 (0.617-1.12), ropivacaine/levobupivacaine: 1.021 (0.774-1.354), respectively. However, a significantly less number of effective analgesia and higher hourly local anesthetic use were observed in the concentration of 0.05% than those of >or=0.1% within each group (P<0.05).</p><p><b>CONCLUSIONS</b>Using patient-controlled epidural analgesia, lower concentrations of bupivacaine, ropivacaine and levobupivacaine with sufentanil produce similar analgesia and motor block and safety for labor analgesia. The analgesic efficacy mainly depends on the concentration rather than the type of anesthetics.</p>

Study on hematopoiesis reconstitution by co-transplant of human bone marrow mesenchymal stem cells and umbilical cord blood CD34(+) cells at different ratios in NOD/SCID mice / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of co-transplant of human bone marrow mesenchymal stem cells (BMMSCs) and umbilical cord blood (UCB) CD34(+) cells on hematopoiesis reconstruction in NOD/SCID mice and to investigate the optimal proportion between the two kind of cells.</p><p><b>METHODS</b>Female NOD/SCID mice were sublethally irradiated by (60)Co gamma-ray and transplanted with BMMSCs and different ratios of UCB CD34(+) cells. From day +3 till day +42 after transplantation, 20 microl peripheral blood (PB) was collected from the retro-orbital plexus of mice weekly, and the variations of WBC and PLT were counted. Mice were sacrificed 42 days after transplantation, and human CD45 positive (huCD45(+)) cells in PB, BM, and spleen were detected by flow cytometry.</p><p><b>RESULTS</b>Compared with transplant of UCB CD34(+) cells alone, co-transplantation of BMMSCs and UCB CD34(+)cells at ratios of 1:1, 5:1 and 10:1, (1) significantly mitigated the decrease range (P < 0.01) and led to the recovery of WBC and platelet in PB one week earlier (P < 0.05), and the difference among the three groups was not statistically significant (P > 0.05); (2) significantly enhanced hematopoietic stem cells (PB, BM and spleen cells) engraftment in recipient mice, and the effect was most pronounced at the ratio of 10:1. huCD45(+) cells in PB, BM and spleen were increased by (2.75 +/- 0.63), (3.51 +/- 0.86) and (5.18 +/- 0.57) fold, respectively (P < 0.01).</p><p><b>CONCLUSION</b>The optimal hematopoiesis reconstruction is achieved by co-transplant of UCB CD34(+) cells and BMMSCs at a ratio of 1:10.</p>

Synergistic effects of arsenic trioxide and proteasome inhibitor bortezomib on apoptosis induction in Raji cell line / 中国实验血液学杂志

The aim of this study was to explore the synergistic effect of arsenic trioxide and bortezomib on apoptosis of Raji cell line. The cells were treated with arsenic trioxide, bortezomib, low-dose arsenic trioxide combined with bortezomib, respectively. The cell viability and proliferative curve were estimated by trypan blue dye exclusion. The cell apoptosis and cell cycle status were analyzed by flow cytometry. The apoptosis related elements such as caspase-3, BCL-2, BAX, JNK2 and IkappaB-alpha, were measured with Western blot. The results showed that compared with cells treated with mentioned above drugs alone, the proliferative potential of cells in combination group was significantly inhibited (p < 0.01), and apoptosis rate markedly increased (p = 0.001), while obvious cell cycle arrest was not observed. On the protein level, the expression of Caspase-3, BAX and IkappaB-alpha increased, while the expression of BCL-2, and JNK2 decreased. It is concluded that low-dose arsenic trioxide combined with bortezomib synergistically induced apoptosis in Raji cell line which may be mediated by inhibiting NK-kappaB and JNK2 signaling.

Hematopoietic reconstitution by co-transplantation of human BM-MSCs and UCB CD34+ cells at various times in NOD/SCID mice / 中国实验血液学杂志

To investigate the effect of co-transplantation of bone marrow derived MSCs and UCB CD34+ cells at different time points on hematopoietic reconstitution, all NOD/SCID mice were sublethally exposed to irradiation of 60Co gamma ray and transplanted with UCB CD34+ with or without MSCs (3 mice per group). Animals were divided into HSC group and MSC+HSC group (M+H group). In HSC group, 1x10(6) UCB CD34+ cells for each mouse were infused within 4-6 hours after irradiation; the M+H group again was divided into 3 subgroups according to infusion sequence of MSCs and HSCs. (A) M+H simultaneously infused group: MSCs and UCB CD34+ cells were infused simultaneously; (B) M+48H group: MSCs were infused within 4-6 hours after irradiation, while UCB CD34+ cells were infused at 48 hours after irradiation; (C) H+48M group: UCB CD34+ cells were infused within 4-6 hours after irradiation, while MSCs were infused at 48 hours after irradiation. In 3 subgroups infused amounts of MSCs and UCB CD34+ cells all were 1x10(6) cells. From the 3rd day after transplantation, 20 microl peripheral blood was collected from the retro-orbital plexus of mice every week until 42th day after transplantation. 42 days after transplantation, mice were sacrificed, and the percentages of human CD45, CD34, CD19 and CD11b in bone marrow, peripheral blood and spleen were detected by FACS. The results showed that (1) Co-transplantation of MSCs and UCB CD34+ cells simultaneously (M+H group) can mitigate the decrease of WBC and platelet levels (p<0.01) in peripheral blood, and accelated the hematopoietic recovery. While co-transplanting MSCs and UCB CD34+ cells at different time points (M+48H or H+48M), the similar effect was not observed (p>0.05). As far as platelets was concerned, the recovery of platelets in M+48H group was lagged behind that in M+H group (p<0.01). (2) Co-transplantation of MSCs at different time points enhanced the engraftment of hematopoietic cells (p<0.05 or p<0.01), compared with transplantation of CD34+ cells alone. The effect of engraftment enhancement was not lineage restriction (p>0.05). It is concluded that the ideal transplantation effect is achieved when MSCs and UCB CD34+ cells were co-transplanted at the same time, these study results provide experimental basis for clinical application of MSCs.

Inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adherence of ovarian cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adhesion ability in vitro of ovarian cancer cells, and to investigate the mechanisms of action.</p><p><b>METHODS</b>MMP-9 antisense oligonucleotides were transfected by lipofectinmin into ovarian cancer cell line HO-8910PM cells expressing MMP-9 induced with fibronectin. RT-PCR, Western blot and gelatin zymography were used to detected MMP-9 expression of mRNA and protein and enzymatic activity. The ability of invasion and migration of ovarian cancer cells was assayed in Transwell cell culture chamber. Cell adhersion assay was carried out in a microculture well pre-coated with fibronectin.</p><p><b>RESULTS</b>MMP-9 expressions of mRNA and protein were significantly decreased in the antisense-transfected cells. Comparing with the control group, the inhibition rate was 34. 8% and 42. 5% , respectively (P <0. 05). Its gelatin enzymatic activity was inhibited. Matrigel invasion assay and Transwell migration assay revealed markedly reduction in invasion and migration for the antisense group. The inhibition rates were 22. 4% and 24. 8% , respectively. The adhesion ability was also reduced. The inhibition rates were 49. 8% and 38. 3% at 60 min and 90 min, respectively.</p><p><b>CONCLUSION</b>MMP-9 down-regulation can significantly inhibit the ability of invasion and attachment of ovarian cells in vitro. MMP-9 may play an important role in invasion and metastasis of ovarian cells and potentially be a molecular target of blocking invasion and metastasis of ovarian cancer.</p>

Report of a case of hybrid acute leukemia with t (12; 22) and literature review / 中华血液学杂志

<p><b>OBJECTIVE</b>To report a hybrid acute leukemia (HAL) patient with t (12; 22) (p13; q12).</p><p><b>METHODS</b>Chromosome specimens were prepared by direct method and/or short-time culture of bone marrow cells. Karyotyping was performed by R-banding technique. Leukemia surface markers were detected by anti-biotin-biotin complex and monoclonal antibodies. Chromosome painting (fluorescence in situ hybridization, FISH) was performed by using whole chromosome 12 and 22 probes labeled with green and red fluorescence, respectively.</p><p><b>RESULTS</b>The clinical and hematological findings were compatible with the diagnosis of HAL. Lymphoid and myeloid markers were positive on the leukemia cells. Karyotype analysis showed that the patient had t (12; 22) (p13; q12) translocation. A reciprocal translocation between chromosomes 12p and 22q was proved by FISH.</p><p><b>CONCLUSIONS</b>t (12; 22) translocation is a rare chromosome abnormality in leukemia. Patients with t (12; 22) had unique clinical, cytogenetic features. This translocation as a cytogenetic marker for poor-prognosis in leukemia needs to be further studied.</p>

Migration and distribution of allogeneic T lymphocytes in organs of graft-versus-host disease mouse model / 中国实验血液学杂志

This study was aimed to investigate the migration and distribution processes of allogeneic donor T lymphocytes in the organs of recipient mice. GVHD model was established by transfusion of the splenocytes of eGFP transgeneic C57BL/6 mice together with born marrow cells harvested from C57BL/6 mice into BALB/c mice underwent 8.0 Gy total body irradiation. The migration and homing of eGFP(+) cells were tracked by stereo-fluorescent microscopy or inverted fluorescent microscopy and flow cytometry. The enzyme linked immunosorbent assay (ELISA) was performed on supernatants from the tissue homogenates to detect the amount of MIP-1alpha. The results indicated that GVHD clinical manifestation and pathological changes of organs appeared on day 8 post transplantation. eGFP-positive donor T cells in recipient organs were observed by inverted fluorescence microscope in frozen section, or by stereo-fluorescence microscopy in living organs, such as liver, spleen, skin, lungs, bowels, and tongue. The highest expression of MIP-1alpha was on day 7 post transplantation in the liver (491.3 +/- 32.1 pg/ml), and day 3 post transplantation in the spleen (881.5 +/- 45.2 pg/ml), respectively (P < 0.05). It is concluded that GVHD was induced by splenocytes of eGFP transgeneic C57BL/6 mice. eGFP(+) cells in the organs can be observed by fluorescent microscopy. In this GVHD model, donor T cells proliferate and infiltrate in liver, skin, bowels, as well as lungs and tongue. MIP-1alpha may be in relation with the infiltration of T lymphocytes in liver and spleen.

Effects of human mesenchymal stem cells and fibroblastoid cell line as feeder layers on expansion of umbilical cord blood CD34(+) cells in vitro / 中国实验血液学杂志

To investigate the effects of human mesenchymal stem cells (MSC) and human fibroblastoid cell line (HFCL) as feeder layer on expansion of umbilical cord blood CD34(+) cells in vitro, (60)Co gamma-ray irradiated MSC and HFCL were used as feeder layer to expand cord blood CD34(+) cells in culture. The efficiencies of MSC and HFCL on expansion of CD34(+) cells in culture with or without cytokines were compared. The results showed that no matter whether cytokines (rhFL, rhSCF, rhTPO) were added, the proliferation of nucleated cells after expansion for 12 days in HFCL group was statistically higher than that in MSC group, i.e. with cytokines (9797 +/- 361)% vs (7061 +/- 418)%; without cytokines (5305 +/- 354)% vs (1992 +/- 247)%, when the cell numbers at day 0 was accounted as 100%), P < 0.01. The proliferation of propagated CD34(+) cells between MSC group and HFCL without addition of cytokines was not statistically different (820 +/- 191)% vs (825 +/- 305)%, P > 0.05. However, in the presence of cytokines, the propagating rate of MSC group was lower than that of HFCL group (939 +/- 212)% vs (1617 +/- 222)%, P < 0.01. MSC was better than HFCL in maintaining the LTC-IC of UCB CD34(+) cells, i.e. the number of CFU-GM colonies in the fifth week was (129.95 +/- 8.73) /10(5) seeded cells vs (89.81 +/- 10.29) colonies/10(5) cells, P < 0.05; with addition of cytokines, the effect was more obvious, i.e. the number of CFU-GM colonies in the fifth week (192.93 +/- 4.95)/10(5) seeded cells vs (90.47 +/- 14.28) colonies/10(5) seeded cells, P < 0.01. MSC mixed with a certain proportion of HFCL facilitated maintaining the LTC-IC of UCB CD34(+) cells. When the proportion was 4:1, the number of CFU-GM colonies was the highest (186.89 +/- 11.11)/10(5) seeded cells, which was higher than that of both 3:2 group [(138.92 +/- 14.84) colonies/10(5) seeded cells] and MSC only group, i.e. (64.63 +/- 6.11) colonies/10(5) seeded cells, both P < 0.01. It is concluded that HFCL is better than MSC in maintaining the expansion of CD34(+) cells and cytokines can enhance this effect, while MSC are stronger than HFCL in maintaining the LTC-IC of UCB CD34(+) cells in vitro. MSC with addition of a certain proportion of HFCL can significantly enhance the efficiency of CD34(+) cell expansion.

RNA interference (RNAi) as novel approach for gene silencing--review / 中国实验血液学杂志

RNA interference (RNAi), a highly conserved evolutionary process of post-transcriptional gene silencing, can be triggered by small interfering RNAs (siRNAs) that mediate sequence-specific mRNA degradation. The article summarized some aspects of the mechanism of RNAi, siRNA design and delivery of siRNAs to mammalian somatic cells. And some hurdles in practice were also discussed.

The protective effect of amifostine on hydroquinone-induced apoptosis in bone marrow / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the protective effect of amifostine on hydroquinone-induced apoptosis of bone marrow mononuclear cells in vitro.</p><p><b>METHODS</b>The mononuclear cells were separated and divided into four groups: blank control, amifostine group, hydroquinone group, amifostine + hydroquinone group. The cell apoptotic rate was examined in separated group at different time point, and apoptosis was detected by HT stain, then cell morphology was observed under fluorescent microscope and DNA fragments was tested by agarose gel electrophoresis. In addition, apoptotic and necrotic rate was detected by flow cytometer.</p><p><b>RESULTS</b>After 10 hour culture, DNA ladder was detected in the hydroquinone group, but not in other groups. The apoptotic rate was not significantly different between amifostine group and blank control group at different culture time (P > 0.05). After 8 - 12 hour culture, the apoptotic rate in amifostine + hydroquinone group was significantly lower than that in the group of hydroquinone alone (P < 0.01). After 18 - 48 hour culture, the necrotic rate in amifostine + hydroquinone group was lower than that in the group of hydroquinone alone (P < 0.05).</p><p><b>CONCLUSION</b>Amifostine can protect cell from hydroguinone-induced bone marrow damage through inhibition on cell apoptosis, and decrease in cell necrosis.</p>