Minimal residual disease (MRD) is the cause of relapse in leukemia,and its dynamic monitoring has been a hotspot yet proven to be difficult.MRD in acute B-lineage lymphoblastic leukemia has its own characteristics,normal bone marrow produces a large number of B precursor cells,easily mistaken for leukemia cells,and there were no specific gene alterations that can be conveniently detected.This paper summarizes the antibody combinations in flow cytometry,timing for surveillance and polymerase chain reaction for gene detection,and provide a reference for B lymphocytic leukemia MRD monitoring.

Laboratory animal welfare is an important guarantee for the accuracy and reliability of experimental results, so it has been paid attention in scientific research .But the situation is not optimistic in the teaching practice .This paper has specially analyzed laboratory animal welfare and ethics in experiment teaching and provided specific measures involved in legislation , spread of animal welfare and implementation of 3R principle to ensure animal welfare .

Objective To investigate the expression and distribution of leptin in pre-implantation embryo,and its effect on mouse embryo development in vitro. Methods Adult NIH female mice were superovulated and then mated with fertile males of the same strain. On the morning of 1st to 4th d of pregnancy,the oviduct and horn of uterus were dissected out and flushed with DMEM/F12 medium adequately. The embryos collected were subjected to the following procedures: ①Total RNA was isolated from embryos of each stage respectively and mRNA expression of leptin was detected by RT-PCR. ②Immunofluorescence staining was performed on blastocysts to analyze the expression and the distribution of leptin protein under laser confocal microscope. ③Eight-cell embryos were cultured in medium containing leptin antibody at various concentrations to observe the formation and hatching of blastocysts. Results ①Leptin mRNA expression was only detected in blastocyst,and no expression was detected in embryos in other stages. Leptin protein was detected in cytoplasma,membrane of trophoblastic cell and inner cell mass,however no expression was detected in cell nucleus. ②Leptin polyclonal antibody significantly inhibited formation of blastocysts (1∶400,P

Objective Uterine epithelial cells were isolated from pregnant mice and cultured in vitro , and exam-ined the effect of CD82 on the expression of integrin αV,β3, E-cadherin and β-catenin in the cells.Methods The uter-ine epithelial cells were primarily isolated from pregnant mouse uterus .The recombinant adenovirus containing mouse CD 82 gene which had been constructed in our lab infected the uterine epithelial cells .Immunocytochemistry was used to detect the protein expressions of integrin αV,β3, E-cadherin and β-catenin in the uterine epithelial cells , which were infected with CD82 adenovirus or not .Results 1.The purity of primary cultured cells was (93.2 ±0.6)%.2.The transfection efficiency of CD82 recombinant adenovirus was ( 92 ±4.5 )%.The adenoviral particles carrying CD 82 gene indeed ex-pressed CD82 gene and protein in the primary uterine epithelial cells after 24 hours or 48 hours.3.The uterine epithelial cells of pregnant mice on d4 expressed integrin αV, β3, E-cadherin and β-catenin.4.In contrast to the control group, when CD82 adenovirus infected cells , the uterine epithelial cells of pregnant mice on d 4 increased the expression of integrinαV,β3 and β-catenin protein, had no significant changes of E-cadherin.Conclusions CD82 may have effect on the ex-pression of integrin αV,β3 and β-catenin in mouse uterine epithelial cells before implantation .

Comparative,medicine is a branch of laboratory animal seiences which also cuts the first edge of the morden medical sciences.This article mainly discussed the origin of comparative medicine,its contributions to life sciences.relationships between it and laboratory animal science,its importance for life science and medical researches.Most of all.the necessarity and feasibity to teach comparative medicine for the undergraduate students were highlighted

Objective:To evaluate the expression of basic fibroblast growth factor (bFGF) in fetal liver stromal cell(FLSC) during hypoxia so as to provide evidences for hematopoietic stromal cell therapy of brain ischemia. Methods: FLSCs from 14.5-day-old mouse fetuses were cultured and passaged. Subconfluent cells of the fifth passage were cultured in hypoxia condition (95% N_(2) + 5% CO_(2) ). bFGF mRNA and bFGF protein were detected at 2,4,8,16 and 32 h of culture by semi-quantitative RT-PCR and Western blot, respectively. Results: bFGF mRNA and bFGF protein were very low in normally cultured cells, but increased significantly under the condition of hypoxia. Expression of bFGF mRNA obviously increased at 2 h of hypoxia culture and reached peak at 16 h, with the peak-value being 5.52 folds that of the control (-[32.86?9.57-]% vs -[5.94?2.65-]%,P

Objective:To evaluate the expression of transforming growth factor-?1 (TGF-?1) in fetal liver stromal cells during hypoxia,in an attempt to provide evidences for cell therapy of brain ischemia. Methods: Cells from 14.5-day-old mouse fetuses were cultured and passaged. Subconfluent cells of the fifth passage were cultured in hypoxia condition (95% N2 + 5% CO2 ). TGF-?1 mRNA and TGF-?1 were detected by semi-quantitative RT-PCR and Western blot,respectively. Results: TGF-?1 mRNA and TGF-?1 levels,which are very low in normally cultured cells, increased significantly in hypoxia-cultured cells. Expression of TGF-?1 mRNA increased from (5.47?2.76)% to (10.38?3.65)% at 4 h of hypoxia(P

Objective To explore the expression and role of LAT 1 in mouse uterus on early placenta formation on day 8 of pregnancy (D8).Methods One hundred and twenty 6-8-week old SPF female Kunming mice were used in this study.Immunohistochemistry was applied to determine the localization of LAT 1 protein in the mouse uterus on D8 of preg-nancy.The ectoplacental cones (EPCs) were dissected out from D8.5 uterus, and then cultured in vitro with different con-centrations of BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, specific antagonist of LAT1) and L-leucine ( substrate of LAT1) to determine the role of LAT1 during the EPC attachment and outgrowth .Results LAT1 protein was highly expressed in secondary decidual zone and also positively expressed in the mouse uterus on D 8.As a specific antago-nist of LAT1, BCH significantly suppressed the ectoplacental cone outgrowth , whereas L-leucine showed no significant effect on it.Conclusions LAT1 is expressed in the mouse uterus during early placenta formation and promotes ectoplacen -tal cone outgrowth , suggesting that LAT1 may promote the trophoblast invasion into maternal decidual tissue , and partici-pates in the early formation of placenta .

Objective To investigate the effect of recombinant adenovirus containing CD82/KAI1 on the expressions of integrin ?5,integrin ?3,MMP-9,E-cadherin and ?-catenin in uterine stromal cells.Methods Uterine stromal cells were isolated from pregnant mice during implantation window and cultured primarily.The recombinant adenovirus containing mouse CD82/KAI1 gene was constructed.Protein expression of integrin ?5,integrin ?3,MMP-9,E-cadherin and ?-catenin were detected by immunocytochemical methods in the cultured cells with or without CD82/KAI1 adenovirus transfection.Results When the cells which were cultured for 48 h were transfected with the recombinant adenovirus vector at a titer of 6.5?1012 pfu/ml,CD82/KAI1 were obviously upregulated in the primary uterine stromal cells.In the uterine stromal cells from pregnant mice on the 4 d after gestation,integrin ?5,E-cadherin,?-catenin and MMP-9 were detected,while,integrin ?3 were not.But in the cells of nonpregant rats,no expression of these 5 proteins was found.Compared with the cells transfected with blank adenovirus,the recombinant adenovirus encoding CD82/KAI1 upregulated the expressions of ?-catenin and MMP-9(P0.05)and no integrin ?3 was detected in the stromal cells of pregnant mice on day 4 of gestation.Conclusion The recombinant adenovirus containing mouse CD82/KAI1 gene is successfully constructed.CD82/KAI1 might have certain effect on the expressions of integrin ?5,MMP-9,E-cadherin and ?-catenin in mouse uterine stromal cells before embryo implantation.

Objective To investigate the expression of the preferentially expressed antigen of melanoma (PRAME) gene in acute myeloid leukemia (AML),and to evaluate its applicability in monitoring minimal residual disease (MRD).Methods Bone marrow specimens were collected from 63 cases of de-novo AML,while 34 samples from 11 patients were tracked for 28 months.The level of PRAME mRNA was measured by real time RT-PCR.Results The PRAME gene expressed in 52.4 ％ (33/63) of de-novo patients,and the positive rate was highest in M3 than that in other subtypes of AML.The expression of PRAME became negative after treatment and increased in the following months before morphology relapse.Conclusion The PRAME gene is highly expressed in AML and could be a useful marker to monitor MRD.