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.

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.

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.

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>

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>

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.

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.

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>

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.

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>

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>

Inhibitory effects of RNA interference on expression of matrix metalloproteinase-9 gene and invasiveness and adhesion in ovarian cancer cells / 中华妇产科杂志

Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.