Distribution and health risk assessment of formaldehyde in non-newly decorated houses in Ningbo, China / 环境与职业医学

Background Formaldehyde is a common air pollutant in residential buildings, and the health risks caused by formaldehyde in residential buildings can not be ignored. Objective This study aims to evaluate the air concentration of formaldehyde in non-newly decorated houses in Ningbo and its possible health risks. Methods A total of 72 houses without any decoration in the past one year in Ningbo were selected by multi-stage random sampling method. From July 2018 to January 2019, the air samples of living rooms and bedrooms were collected and their temperature and humidity were also measured. The concentrations of formaldehyde were detected by AHMT method according to Standred method for hygienic examination of formaldehyde in air of residential areas — Spectrophotometric method (GB/T 16129—1995) , the health risk assessment model of U.S. Environmental Protection Agency was used to evaluate the non-carcinogenic risk and carcinogenic risk of formaldehyde, and Monte Carlo simulation was used for sensitivity analysis. Results The median (P25, P75) of formaldehyde concentration in the 72 houses was 0.019 (0.012,0.026) mg·m−3. Only one house showed a formaldehyde concentration that exceeded the national standard in the living room, and the total qualified rate of formaldehyde concentration was 98.61%. The median (P25, P75) of formaldehyde concentration in the bedroom was 0.019 (0.011, 0.031) mg·m−3, which was higher than that in the living room, 0.015 (0.010, 0.024) mg·m−3, and the difference was statistically significant. The median and 90th percentile of non-cancer risk (hazard quotient, HQ) of the 72 houses were 1.35 and 2.80, respectively, and the proportion of the houses with HQ>1 was 62.50%. The median and 90th percentile of cancer risk (CR) of the 72 houses were 1.12×10−4 and 2.32×10−4, respectively, and the proportions of the houses with CR>1×10−6, CR>1×10−5, and CR>1×10−4 were 100.00%, 100.00%, and 54.20%, respectively. After using Monte Carlo simulation, the median (90th percentile) of non-carcinogenic risk was reduced to 0.91 (1.94), where the median was lower than the national limit, and the proportion of samples with HQ>1 was 44.73%; the carcinogenic risk was reduced to 7.52×10−5 (1.79×10−4), and the proportions of samples with CR>1×10−6, CR>1×10−5, and CR>1×10−4 were 100.00%, 98.96%, and 34.37%, respectively. Conclusion The concentration of formaldehyde in non-newly decorated houses in Ningbo basically meets the national requirements, but it is still necessary to pay attention to the non-carcinogenic risk and carcinogenic risk caused by indoor formaldehyde, among which the carcinogenic risk is more important. Residents should prevent the harm of formaldehyde from its source by considering clean decoration materials and environmentally friendly furniture.

Epidemiological characteristics of hospitalized children with burn in Ningbo / 预防医学

Objective@#To analyze the epidemiological characteristics of hospitalized children with burn in Ningbo from 2013 to 2018,and to provide evidence for developing intervention strategies. @*Methods@#The pediatric burn cases,discharged from the Ningbo Women & Children's Hospital from 2013 to 2018,were registered using the Ningbo Hospitalized Injury Monitoring Report Card,and their distributions of time,places,groups,involved body parts and prognosis were analyzed. @*Results@#There were 3 862 pediatric burn inpatients included in this study,with 2 977（77.08%）cases aged 1-3 years and 2 532（65.65%）nonlocal cases. About 898（23.25%）cases occurred during 9:00-12:00 a.m. and 1 833（47.46%）cases occurred during 17:00- 21:00 p.m. Burns predominantly occurred at home,with 3 810（98.65%）cases. The top three body regions involved were multiple regions,lower limbs and upper limbs with 1 820（47.13%）,835（21.62%）and 541 cases（14.01%）. The proportions of involving multiple regions declined with age（Psingle<0.05）. The proportion of involving multiple regions in nonlocal children was higher than that of local children（P<0.05）. @*Conclusion@#Burn is one of the leading causes of child injury-related hospitalization in Ningbo. Home is the main burn scene. Nonlocal and 1-3 year-old children were especially at high risk of burns.

Brain derived neurotrophic factor enhances the role of mesenchymal stem cells in inhibiting follicular helper T cells / 中华血液学杂志

Objective@#To investigate the effect of brain derived neurotrophic factor (BDNF) on mesenchymal stem cells (MSC) inhibiting follicular helper T cells (Tfh cells).@*Methods@#The contents of indoleamine 2,3-dioxygenase (IDO), IL-10, TGF-β and IL-21 in MSC culture supernatant were detected by ELISA; The peripheral blood of healthy volunteers were collected, and lymphocyte in peripheral blood was separated by human lymphocyte separation solution; Co-cultures of MSC and lymphocyte were performed by Transwell chamber, and the proportion of CD4+CXCR5+ Tfh cells and their subtypes were detected by flow cytometry.@*Results@#①The concentrations of IL-10, TGF-β, and IDO in the supernatant of BDNF group (BDNF-stimulated MSC) were higher than those of the control ones (adding PBS with the same volume) [IL-10: (42.1±4.4) ng/ml vs (19.3±2.1) ng/ml, t=4.761, P=0.009; TGF-β: (13.9±1.7) ng/ml vs (5.3±0.6) ng/ml, t=5.129, P=0.008; IDO: (441.3±56.9) ng/ml vs (226.7±37.6) ng/ml, t=3.130, P=0.035]; ②The comparisons between BDNF (co-culture of lymphocyte and BDNF-stimulated MSC) and MSC groups (co-culture of lymphocyte and MSC) were detailed as of follows: the proportion of CD4+ CXCR5+Tfh cells were lower [(3.37±0.21)% vs (6.51±0.27)%, t=9.353, P<0.001], the proportion of CD4+ CXCR5+CXCR3+ CCR6- Tfh cells were higher [(41.14±2.04)% vs (26.72±2.57)%, t=4.383, P=0.012], CD4+CXCR5+CXCR3-CCR6- Tfh2 cells and CD4+CXCR5+CXCR3-CCR6+ Tfh17 cells were lower [Tfh2: (30.16±5.38)% vs (43.26±4.11)%, t=4.426, P=0.012; Tfh17: (15.61±1.52)% vs (22.32±0.72)%, t=4.202, P=0.014], the proportion of CD4+CXCR5+Foxp3+ Tfr cells were higher [(4.95±0.22)% vs (2.32±0.16)%, t=10.241, P<0.001], the concentration of IL-21 in the lymphocyte supernatant was lower [(0.28±0.03) ng/ml vs (0.85±0.08) ng/ml, t=6.675, P=0.003].@*Conclusion@#BDNF could enhance the inhibitory effect of MSC on Tfh cells through inhibiting the increasing of Tfh cells and the differentiations of Tfh2 and Tfh17 cells.

Survival analysis of patients with high risk refractory lymphoma treated by autologous hematopoietic stem cell transplantation / 重庆医学

Objective To investigate the prognosis and survival situation of the patients with high risk refractory lymphoma treated by autologous hematopoietic stem cell transplantation .Methods The follow up data in 110 patients with high risk refractory lymphoma treated by autologous hematopoietic stem cell transplantation in our hospital were retrospectively analyzed .The survival and prognosis factors of the patients after autologous hematopoietic stem cell transplantation were analyzed by using the Kaplan-Meier survival analysis and Cox proportional hazard regression analysis .Results The median survival time was 39 .4 months in 110 cases ,the 3‐year overall survival rate (OS) and progression free survival rate (PFS) were 80 .9% and 76 .4% respectively .The pa‐tients achieved CR status before transplantation(P=0 .016) and consolidation therapy after transplantation (P=0 .006) were the favorable prognostic factors of the patients undergoing transplantation .The prognosis in the patients with high LDH values ,IPI score>2 and bone marrow infiltration and HBV infection were poor(P<0 .05) .Conclusion Autologous hematopoietic stem cell transplantation can improve the long‐term survival rate in the patients with high risk refractory lymphoma .

Association of microRNA-related genes (DROSHA, DICER1 and GEMIN4) polymorphisms with T-cell lymphoma prognosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the association of micoRNA-related genes DROSHA single nucleotide polymorphisms (SNP) rs10719 and rs6877842, DICER1 rs3742330and GEMIN4 rs3744741 with prognosis of T-cell lymphoma.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to determine the genotypes of the above 4SNPs and their associations with complete remission (CR) rate and overall survival (OS) in 163 patients with TCL.</p><p><b>RESULTS</b>Patients carrying the rs6877842 CG genotype had a significantly higher CR rate compared with those carrying the CC genotype (OR=0.07, 95% CI 0.01-0.72, P=0.026); the same for patients carrying the DICER1 rs3742330 GG genotype compared with those carrying the GA genotype (OR=0.15, 95% CI 0.02-0.97, P=0.047) or the AA genotype (OR=0.11, 95% CI 0.02-0.71, P=0.020). In addition, patients with the DICER1 rs3742330 GG genotype had a significantly improved OS compared with those carrying the GA (HR=9.02, 95% CI 1.22-66.92, P=0.031) or AA genotype (HR=8.77, 95% CI 1.19-64.67, P=0.033). The other two SNPs of rs10719 and rs3744741 had no significant association with CR or OS.</p><p><b>CONCLUSION</b>DROSHA rs6877842 and DICER1 rs3742330 were independent factors for TCL CR, and DICER1 rs3742330 was also an independent prognostic factor for TCL OS.</p>

Study on expression of SENP1 and c-myb gene in patients with acute lymphoblastic leukemia / 重庆医学

Objective To investigate the SENP1 and c-myb gene expression and their correlations in bone marrow specimens in the patients with acute lymphoblastic leukemia (ALL ) to provide the basis for expounding the role ,mechanism and prognosis of SENP1 and c-myb in ALL .Methods 31 patients diagnosed with ALL (22 cases of B-ALL ,1 case of T-ALL and 8 cases of uncate-gorized ALL ;6 cases in the low/medium risk group ,25 cases in the high risk group) and 31 patients with proliferative bone marrow and hyperplastic anemia diagnosed by the morphology were taken as the control group .The real-time PCR and immunocytochemical staining(SP method) were adopted to detect the mRNA and protein expressions of SENP1 and c-myb in the bone marrow specimens of the ALL patients and the control group .Results The expression of SENP1 and c-myb were both increased in the bone marrow specimens and smears of ALL patients ,which showed the statistical difference compared with the control group (P< 0 .05) ,the Pearson correlation analysis found that the high expression of SENP1 and c-myb had correlation .The expression of SENP1and c-myb in the low/medium risk group were lower than that in the high risk group ,but the difference had no statistical significance . Conclusion The high expression of SENP1 and c-myb exists in the bone marrow specimens of the ALL patients ,SENP1 and c-myb could possibly have the correlation with the occurrence and development of ALL ;but now the differences of SENP1 and c-myb ex-pression among different risk groups of ALL patients are yet to be proven .

Clinical observation of arsenic trioxide plus all trans retinoic acid versus ATRA combined with chemotherapy treatment on acute promyelocytic leukemia / 重庆医学

Objective To observe the efficacy of arsenic trioxide(ATO) combined all trans retinoic acid (ATRA) versus cytara‐bine (Ara‐C) combined ATRA in the treatment of acute promyelocytic leukemia(APL) .Methods We enrolled 65 patients in our department during the period between January 2002 and August 2008 ,and they were randomly assigned to receive ATRA combined ATO (treatment group ,n= 27) or ATRA combined DA ,HA ,NA which were major of Ara‐C (control group ,n= 38) .Then observe the differences of between the two groups ,such as complete remission(CR) ,the time to complete remission ,overall survival(OS) ,e‐vent free survival(EFS) ,the 5 years disease free survival (DFS) and adverse reactions .Results The CR rate of treatment group (ATRA + ATO) and control group (chemotherapy + ATRA) was 81 .48% and 68 .42% ,respectively ,and the time to complete re‐mission was (28 .50 ± 3 .97)d and (30 .56 ± 2 .39)d ,respectively ,showed that there was no statistical difference between the two groups ( P > 0 .05 ) .The 5 years DFS of the CR patients in the two groups was 51 .9% (ATRA + ATO ) and 50 .0%(Chemotherapy + ATRA) ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .The 5 years EFS of the CR patients in the two groups was 48 .1% and 39 .5% ,respectively ,showed that there was no statistical difference between the two groups(P> 0 .05) .The 5 years DFS of the patients in the two groups was 55 .6% and 67 .6% ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .Bone marrow suppression in the treatment group was significantly lower than in the control group(P< 0 .05) .Conclusion ATRA + ATO can prolong the CR rate ,OS ,EFS and 5 years EFS of newly diagnosed APL patients .ATRA combined with chemotherapy has similar efficacy ,ATRA + ATO has lower bone marrow suppression than the ATRA combined with chemotherapy ,thus may reduce the risk of early death .

Itraconazole application for prevention of fungal infection in patients receiving allogeneic hematopoietic stem cell transplants / 重庆医学

Objective To evaluate the clinical effect of itraconazole in prevention of invasive fungal infections in allogeneic hema-topoietic stem cell transplantion .Methods In this retrospective study ,110 patients receiving allogeneic hematopoietic stem cell transplants were administed itraconazole or fluconazole for prevention of fungal infection .The occurrence and prognosis of invasive fungal infection ,and the side effect of both pyrroles were observed .Results Proven and probable invasive fungal infections occurred in 5 of 69 itraconazole recipients(7 .2% ) and in 8 of 41 fluconazole recipients(19 .5% ) during the first 180 days after transplanta-tion ,the difference had statistical significance(P<0 .05) .The fatality rate related to fungal infection had no statistical difference be-tween the two groups(2 .9% vs .7 .3% ) .The occurrence of itraconazole adverse reactions were more than fluconazole (26 .9% vs . 7 .0% ) ,and both itraconazole and fluconazole were well tolerated .Conclusion Itraconazole significantly reduces the incidence of inva-sive fungal infection in the patients receiving allogeneic hematopoietic stem cell transplants ,and it is a effective and safe prophylaxis .

Efficacy of arsenic trioxide for acute promyelocytic leukemia: a systematic review and meta-analysis / 中西医结合学报

Objective: To systematically review the efficacy and safety of arsenic trioxide (ATO) in treatment of acute promyelocytic leukemia (APL). Methods: The Cochrane Library (Issue 1, 2009), Cochrane Central Register of Controlled Trials (from 1970 to January 2009), MEDLINE (from 1978 to October 2008), EMBASE (from 1950 to March 2009), Chinese Biological Medical Literature Database (from 1978 to December 2008), China National Knowledge Infrastructure (CNKI, from 1994 to December 2008), and China Medical Academic Conference Database (from 1994 to December 2008) were electronically searched. We also searched the Meta-Register of controlled trials, Conference Proceedings of American Society of Hematology (from 1946 to December 2008) and Conference Proceedings of American Society of Clinical Oncology (from 1946 to December 2008) on the internet for grey literature. The related journals in the library of Third Military Medical University were hand-searched. The randomized controlled trials (RCTs) of ATO in treatment of APL were included. We adopted complete remission, overall survival rate, disease free survival rate, time to complete remission, relapse rate, mortality and adverse reactions as outcome indicators. Data were entered and analyzed with the Cochrane review manager software 5.0 (RevMan 5.0). Results: After merger of the included trials, five eligible RCTs with 328 cases were included. All the RCTs focused on the comparison of all-trans retinoic acid (ATRA) plus ATO regimen with ATRA monotherapy. Meta-analysis showed that the effect indexes for time to complete remission, two-year disease free survival rate, relapse rate, incidence of edema and incidence rate of QT interval prolongation were -1.20 [-1.68, -0.72], 8.64 [1.66,45.00], 0.21 [0.09,0.47], 4.16 [1.46,11.79] and 22.10 [2.75,177.49], respectively. The influences on other outcome indicators such as complete remission and leukocytosis were statistically non-significant. Conclusion: ATO can prolong disease free survival and reduce the time to complete remission and relapse rate of newly diagnosed APL patients, and increase the incidence of edema and prolongation of corrected QT interval during the treatment. Due to limitation of the included trials, this conclusion needs to be validated by further studies.

Arsenic trioxide in combination with all-trans retinoic acid for acute promyelocytic leukemia: a systematic review and meta-analysis / 中西医结合学报

The studies have demonstrated that arsenic trioxide (ATO) in combination with all-trans retinoic acid (ATRA) takes effects in treatment of acute promyelocytic leukemia (APL) through different underlying mechanisms. This has established the molecular foundation of ATO plus ATRA therapy. Currently, ATO plus ATRA has also been widely used in clinical practice.

Effects of recombinant antisense c-myc adenovirus in induction of differentiation of HL-60 cells / 第三军医大学学报

Objectives To evaluate the potential of antisense c myc gene therapy for acute promyelocytic leukemia by investigating the biological effects and molecular mechanisms in induction of differentiation of HL 60 cell line using recombinant antisense c myc adenovirus (Ad AS c myc). Methods Cultured HL 60 cells treated with Ad AS c myc or Ad LacZ with polybrene and protamine sulfate were analyzed by X gal staining, morphology, MTT, flow cytometric analysis, RT PCR, and immunocytochemical techniques in vitro . Results HL 60 cells could be transfected effectively by Ad LacZ+protamine sulfate (79.8%). The level of c myc transcription and the version could be strongly inhibited by Ad AS c myc in the transfected HL 60 cells. Ad AS c myc could strongly inhibit the cell growth in HL 60 cells (51%). Ad AS c myc could reduce the ratio of nuclear/cytoplasm, and increase the activity of peroxidase in HL 60 cells. Ad AS c myc could lead to the blocking of G 0/G 1 phase in HL 60 cells. Ad AS c myc could also increase the expression of c fos in HL 60 cells. Conclusion The expression of Ad AS c myc can inhibit the growth and induce differentiation of HL 60 cells in vitro . The biological effects of Ad AS c myc may be closely associated with the activity of peroxidase and c fos gene in HL 60 cells. Ad AS c myc is of clinical potential in gene thera py for acute promyelocytic leukemia.

Expressions and activation of JAK2 and STAT5 during the process of differentiation of human cord blood CD34~+ cells into dendritic cells / 第三军医大学学报

Objective To understand the roles of JAK STAT pathway in the process of differentiation of human cord blood CD34 + hematopoietic stem cells into dendritic cells (DCs) by detecting the expressions and activation of JAK2 and STAT5. Methods CD34 + hematopoietic stem cells isolated from human umbilical cord blood and cultured for two weeks were induced to differentiate into DCs in vitro . Total cellular JAK2 and STAT5 and tyrosine phosphorylated protein stimulated by granulocyte/macrophage colony stimulating factor (GM CSF) at different time points (0, 7, and 14) during DC differentiation were detected by Western blotting. Results The amount of JAK2 protein was similar at 0, 7, and 14 d without GM CSF stimulation. With the differentiation of cells into DCs, the amount of tyrosine phospho JAK2 induced by GM CSF increased markedly. Both total cellular and tyrosine phospho STAT5 expression increased markedly during DC differentiation. Maximal tyrosine phospho STAT5 expression was later than JAK2. Conclusion JAK STAT pathway may take part in the signal mechanism of DCs differentiation from CD34 + hematopoietic stem cells stimulated by GM CSF.

Arsenic trioxide vs all-trans retinoic acid to treat acute promyelocytic leukemia: a systematic comparative review / 第三军医大学学报

Objective To compare the efficacy and safety of arsenic trioxide ( ATO) with all-trans retinoic acid ( ATRA) for the treatment of acute promyelocytic leukemia ( APL) . Methods We searched the database of Cochrane Library ( Issue 1,2009) ,CENTRAL ( 1970 to 2009) ,Medline ( 1978 to 2008) ,EMBASE ( 1950 to 2009) ,CBM ( 1978 to 2008) ,CNKI ( 1994 to 2008) and CMAC ( 1994 to 2008) . We also searched the Meta register,Conference Proceedings of American Society of Hematology ( 1946 to 2008) and American Society of Clinical Oncology ( 2004 to 2008) on the internet for grey literature. We had searched the related journals in the library of Third Military Medical University,too. We included randomized controlled trials which compared ATO with ATRA for the treatment of APL. We adopt complete remission rate,overall survival rate, disease-free survival rate,time to complete remission,relapse rate,mortality and adverse reactions as result indicators. Data were entered and analyzed with the Cochrane review manager software ( Revman 5. 0) . Results Four eligible randomized controlled trials ( RCTs) were included ( n =243) . All the RCTs were methodologically graded as B. They all are focusing on the comparison of ATO monotherapy with ATRA monotherapy in treating newly diagnosed APL patients. Meta analysis showed that effect index for complete remission,2-year disease-free survival,time to complete remission,relapse rate and mortality was 0. 96 ( 0. 50,1. 86) ,2. 76 ( 0.71,10.66) ,-1.30 d ( -1.83,-0.78) ,0.86 ( 0.45,1.63) ,and 1.15 ( 0.45,2.95) ,respectively. All indicated no statistically significant difference. Effect index for incidence of liver dysfunction was 3. 03 ( 1. 25, 7. 37) ,which showed statistically significant difference between ATO group and ATRA group. Conclusion ATO is not superior to ATRA in treating newly diagnosed APL patients regarding complete remission,diseasefree survival rate,time to complete remission,relapse rate and mortality. What is worse,it will increase the incidence of liver dysfunction during treatment. Due to limitation of included trials,this conclusion need to be validated by further studies.

The mechanism of apoptosis of acute promyelocytic leukemia cells induced by arsenic trioxide / 解放军医学杂志

Objective To investigate the effects and the gene expression of mitochondria on arsenic trioxide-induced apoptosis of acute promyelocytic leukemia (APL) NB4 cells. Methods NB4 cells were treated with As2O3. Fluorescent microscopy and flow cytometry were used and mitochondria membrane potential detection and RT-PCR were performed to observe the NB4 cell apoptosis, growth inhibitory effect and mitochondrial transmembrane potentials. Mitochondria genome primers were designed, and the expression of mitochondria genome at gene and protein levels was studied. Results Induced by As2O3, the NB4 cells showed typical morphological changes of apoptosis with a significant growth inhibitory effect. The ratios of NB4 cells apoptosis were 5.02%, 6.40%, 28.40% and 33.34%, respectively, when treated with As2O3 in concentrations of 0.5?mol/L, 1?mol/L, 2?mol/L and 3?mol/L. When treated with As2O3 in 0.5?mol/L, 1?mol/L, 2?mol/L, 4?mol/L and 8?mol/L at 48h, the mitochondria potential of NB4 cells was decreased by 12.8%, 21.6%, 66.9%, 83.7% and 83.8%, respectively. After the NB4 cell apoptosis was induced by As2O3, RT-PCR assay was used to detect the expression of 13 genes of mitochondria genome. The expression of COX2 gene was down-regulated in this process, while no change was found in the expression of other 12 genes. Conclusions As2O3 has shown to exert a significant effect on promoting apoptosis and growth inhibition on APL cells. The apoptotic effect induced by As2O3 on NB4 cells is closely related to a decrease of mitochondrial membrane potential. The expression change in the mitochondria gene COX2 is involved in the As2O3-induced apoptosis of NB4 cells.

Relationship between genetic polymorphism of methylenetetrahydrofolate reductase and the risk of acute lymphocytic leukemia / 解放军医学杂志

Objective To investigate the relationship between genetic polymorphism of methylenetetrahydrofolate reductase (MTHFR) and the risk of acute lymphocytic leukemia (ALL). Methods Eighty-three patients with ALL and a cohort of 83 matched healthy objects were included, and DNA was extracted from their peripheral blood. PCR-RFLP was used to determine the genotypes of MTHFR C677T and A1298C. The adjusted odds ratio (OR) and 95% confidence interval (CI) were calculated using unconditional logistic regression model. Results It was found that the frequency of the MTHFR C677T TT genotype among patients was significantly different from that among control objects (P=0.008). The MTHFR C677T TT genotype had an increased risk of ALL compared with that of 677CC genotype (OR=3.229, 95%CI: 1.328-7.847, P=0.01). No significant association between the MTHFR C677T CT genotype or A1298C polymorphism and the risk of leukemia. Conclusion The present findings suggest that 677C→T polymorphism in MTHFR may be a genetic susceptibility factor for acute lymphocytic leukemia.

The influence of expression level of c-myb on haemopoietic commitment,differentiation and the expression of haemopoietic related genes / 解放军医学杂志

Objective To establish the cell models of c-myb+/+ and c-myb+/-ES in vitro by gene targeting with ES cell culture system, with the aim to examine the detailed role of the transcription factor c-myb in haemopoietic commitment and differentiation. Methods Haemopoietic differentiation was initiated by seeding ES cells in LIF-free methylcellulose medium. The embryoids of c-myb+/+ and c-myb+/-ES were analyzed by methylcellulose colony assay and real-time PCR, the formative process of embryoids and the expression of relative genes were compared in each haemopoietic system at different differentiation stages and procedures. Results The formative process of embryoids was similar for both c-myb+/+ and c-myb+/-ES cells, but the size and frequency of EBs were reduced in the case of the c-myb+/-cells. Similar kinetics gain existed for the formation of CFU-Es in both groups, but the number in c-myb+/-group was less than that of c-myb+/+ group, and the colonies were generally smaller. BFU-E was first detectable on day 7, and the peak value emerged on day 10 in both groups. Similar kinetics gain existed for the formation of CFU-M in the two groups, but the number was larger in c-myb+/-group than that in c-myb+/+ group, while the number of CFU-GM in c-myb+/-group was less than that in c-myb+/+ group. Real-time PCR analysis showed no changes on the gene expressions of ?-globin, ?-globin, GATA3, CD34, Lys, c-fms and c-mpl in both groups. Conclusions Sub-level expressions of c-myb (55%) and c-myb+/-are sufficient to allow progenitor to expand, but they throw a negative influence on the terminal differentiation. Sub-level expression of c-myb may influence the erythropoiesis and granulocytic development, but throw no influence on the precursor cells to differentiate into macrophages. The expressive levels of c-myb have no effect on the expressions of ?-globin, ?-globin, GATA3, CD34, Lys, c-fms and c-mp.

Roles of c-myb in haemopoietic differentiation and commitment of embryonic stem cells in vitro / 第三军医大学学报

Objective The c-myb is an important transcription factor in early haemopoietic system development and differentiation. We wish to use ES cell culture system by gene target getting cell models of c-myb+/+ and c-myb-/- ES in vitro in order to examine the detailed roles of the transcription factor c-myb in haemopoietic commitment and differentiation. Methods Haemopoietic differentiation was initiated by seeding ES cells in methylcellulose medium in the absence of LIF. The embryoid bodies of c-myb+/+ and c-myb-/- ES were analyzed by methylcellulose colony assay and real-time PCR to compare the formation of each haemopoietic system on different differentiation stage and procedure and relative gene expression. Results The formation of embryoid bodies was similar for both c-myb+/+ and c-myb-/- ES cells with the exception that the size and frequency of EBs reduced in the case of the c-myb-/- cells. The number of embryoid bodies increased more markedly in c-myb+/+ than that in c-myb-/-. Erythroid progenitors (CFU-E) were first present in c-myb+/+ group at 6 d after in vitro differentiation, and their number reached the peak at 8 d. Similar kinetics were seen for the formation of CFU-Es in c-myb-/- group, but their number was lower than that in c-myb+/+ group, and the colonies were generally small. CFU-M were first detectable at 7 d, and the peak levels were present at 7 d in c-myb+/+ group. Similar kinetics were seen for the formation of CFU-M in c-myb-/- group, but their number was lower than that in the c-myb+/+ group. CFU-GM was first detectable at 7 d, and the peak levels were present at 12 d in c-myb+/+ group. CFU-GM was not detected in c-myb-/- group. Real-time PCR analysis showed that there was no change in the gene expressions of ?-globin, zeta-globin, Lys, and C-fms in the two groups of c-myb+/+ and c-myb -/-. Conclusion Low levels of c-myb are suff-icient to allow progenitor expansion, but progression of progeniters towards terminal differentiation is significantly altered. Low levels of c-myb can influence erythropoiesis development, but precursor cells capable of differentiating into macrophages are present in haemopoietic commitment. The levels of c-myb can not change the gene expressions of ?-globin, zeta-globin, Lys, and C-fms.

PREPARATION OF MONOCLONAL ANTIBODIES AGAINST HUMAN VASCULAR ENDOTHELIAL CELLS AND THEIR POSSIBLE APPLICATION IN VASCULAR ENDOTHELIAL CELL TYPING / 中国免疫学杂志

Human vascular endothelial cells(VEC)were isolated from umbilicalyeins by trypsin digestion.Spleen cells from BALB/c mice having been im-munized with the isolated endothelial cells were fused with the mousemyeloma cell line SP2/0.After screening and subcloning,three differentmonoclonal antibodies(EC1,EC2,EC3)against human umbilical VEC wereobtained.These antibodies were proved to belong to immunoglobulin sub-classes(IgG_2,IgG_3,IgG_2).EC1 was shown to bind strongly to the vascularendothelial cell membrane,but not to react with peripheral lymphocytes andmonocytes in microcytotoxicity test.We also explored the possible applica-tion of the monoclonal antibodies in VEC typing.

Studies with an oligonucleotide microarray on the changes of gene expression related with NB4 apoptosis induced by arsenic trioxide / 解放军医学杂志

Objective Gene microarray technology was used to investigate the differential gene expression of apoptosis related genes in NB4 cells induced by arsenic trioxide. Methods The databases of Evntrez and Human IPI were searched with "apoptosis or apoptotic" as the key words, and 1384 apoptosis related genes were found after the redundant genes were eliminated by chromosomal localization. The probes of these genes were designed using OligoArray 2.0, and then analyzed by BLAST. All the probes were immobilized on the glass slide, which were used as oligonucleotide microarray. After NB4 cells were treated with 2umol/L As2O3 for 48h, the total RNA were extracted. cDNAs of control group and test group were fluorescently labeled with Cy3 and Cy5, respectively, by RT-PCR. The fluorescent samples were hybridized with an oligonucleotide microarray containing 1384 apoptosis related genes to search for the differentially expressed genes in the cells with or without As2O3 treatment. The hybridization signals were scanned by oligonucleotide microarray, and then the fluorescent intensity of Cy3 and Cy5 and the ratio of two fluoresceins were analyzed using certain software. Then the differential expressed genes were analyzed after As2O3 treatment, in which the most distinctly differential expressed genes were chosen as targets, and through PCR amplification and gel electrophoresis the above genes were verified. Results There are 4 genes up-regulated and 12 genes down-regulated in expression in NB4 cells after 48h treatment with 2umol/L As2O3, which were in accordance with the results of RT-PCR and oligonucleotide microarray. Conclusion Differential gene expression in NB4 cells was induced by As2O3 treatment. These differentially expressed genes, with relation to signal transduction, transcription regulation, cell cycles, oxidation response, protein translation and cell differentiation, may play an important role in NB4 cell apoptosis.