Objective To evaluate the significance of dual-color interphase fluorescence in situ hybridization (FISH) in the engraftment estimation and minimal residual disease (MRD) monitoring after sex-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method The fluorescence signal of samples from 35 cases in different periods after sex-mismatched allo-HSCT were detected by interphase FISH using chromosome enumeration probes (CEP) X and Y. Results All of 35 patients had been determined to obtain engraftment after allo-HSCT. When the disease relapsed,FISH showed that the percentage of donor chromosomes was decreased and when the disease got remission,the percentage of donor chromosomes increased. When conventional cytogenetics showed 100 % XX or 100 % XY,FISH showed different percentage of host chromosomes.Conclusions The test of dual-color interphase FISH is reliably sensitive and simple for engraftment evaluation and MRD monitoring post HSCT. It is a good complement method to cell morphology and traditional karyotype analysis.

BACKGROUND:There are less studies addressing whether bone marrow mesenchymal stem cells from systemic lupus erythematosus patients are different from healthy people.
OBJECTIVE:To compare the multi-differentiation capacity of bone marrow mesenchymal stem cells isolated from systemic lupus erythematosus model mice and normal control mice.
METHODS:Bone marrow mesenchymal stem cells of systemic lupus erythematosus model mice and C57BL mice were isolated and cultured fol owed by osteogenic and adipogenic differentiations, respectively. Differentiation abilities of two kinds of mouse bone marrow mesenchymal stem cells were observed.
RESULTS AND CONCLUSION:Passaged bone marrow mesenchymal stem cells from C57BL mice were long spindle-shaped and evenly distributed, while bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice showed slow growth and were relatively smal er than those from C57BL mice. After osteogenic induction, the amount of calcium salt and calcium nodules were significantly less in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice than from normal control mice. PCR detection showed that expressions of Runx2, alkaline phosphatase and osteocalcin were also significantly decreased in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice. After adipogenic induction, the number of lipid droplets in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice was significantly less than the control group, and PCR detection also showed significantly decreased expression of adipogenic genes, including PPARγ2 and lipoprotein lipase. These findings suggest that the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice exhibit lower osteogenic and adipogenic capacities than those from normal C57BL mice, and also have the impaired multi-differentiation ability.

BACKGROUND:Tree shrew is a representative between insectivore and primates, has a high degree of evolution, is more inexpensive primates, has high use of medical biology, and has been attached by scholars. OBJECTIVE:To detect whether the commonly used secondary antibodies have immune response with tree shrew serum. METHODS:Western blot assay and enzyme linked immunosorbent assay were utilized to detect whether the tree shrew serum had cross-reacts with anti-rabbit, anti-goat, anti-human, anti-mouse, anti-rat, and anti-monkey secondary antibodies. RESULTS AND CONCLUSION:Western blot assay results indicated that tree shrew serums did not react with anti-rabbit, anti-goat, anti-human, anti-mouse, and anti-rat secondary antibodies and had cross reaction with anti-monkey secondary antibody. Enzyme linked immunosorbent assay results also indicated that tree shrew serums were cross-reactive with anti-monkey secondary antibody, but did not have cross-reactivity with the other secondary antibodies. Above data confirmed that the usual y soled secondary antibody cannot be used to immunoassay with tree shrews IgG. Only anti-monkey secondary antibody has cross-react with tree shrew serum. It is necessary to prepare anti-tree shrew IgG monoclonal and polyclonal antibodies. When no antibody is readily available at present, anti-monkey secondary antibody can be used to substitute detection, and can be widely applied in the study of tree shrew models of disease.

Objective To evaluate the MR perfusion imaging in measuring the hemodynamics of liver.MethodsCT and MR perfusion imaging were performed in livers of 13 pigs.The hepatic artery perfusion(HAP),portal vein perfusion(PVP),total hepatic blood flow(THBF),portal vein perfusion index(PVI),distribution volume(DV)and mean transit time(MTT)were calculated and compared respectively.Hemodynamics of the liver from two kinds of imaging was compared by paked t-test,and the relativity was analyzed by Pearson correlation analysis.ResultsIn CT and MR perfusion imaging of the 13 pigs:the hepatic artery perfusion(HAP)were(37.7±7.38)and(35.80±7.31)ml-min-1·100ml-1.the portal vein perfusion(PVP)were(123.16 35.89)and(121.40±36.81)ml·min-1·100 ml-1.the total hepatic blood flow(THBF)were(160.88±37.05)and(157.21±38.71)ml·min-1·100 ml-1.the portal vein perfusion index(PVI)were(75.95±5.21)%and(76.63±5.24)%,the distilbution volume(DV)were(35.10±11.17)%and(41.03±10.06)%。The mean transit time(MTT)were(14.08±1.40)and(14.94±1.32)s.There were no statistical differences for HAP,PVP,THBF and PVI between CT and MR perfusion imaging(t=1.263,0.926,1.225,-1.115,P＞0.05).There were no statistical differences for DV and MTT(t=-3.345,-3.200,P＜0.01).Analysis of these two kinds of imaging showed the correlation coefficient of PVP,THBF and PVI were hishly related(r＞0.85,P＜0.01).Correlation coefficient of HAP,DV and M1T were moderately related(r＞0.70,P＜0.01).ConclusionsMR perfusion Call quantify the blood flow parameters of liver accurately.It is close to that of the CT perfusion imaging.

AIM:To explore the effects of confluent endothelial progenitor cells (EPCs) derived from young and aged rats on the phenotype conversion, proliferation and migration of vascular smooth muscle cells ( SMCs) .METH-ODS:Mononuclear cells were obtained from the bone marrow of young (1~2 month old) and aged (19 to 26 month old) Sprague-Dawley rats and cultured with medium DMEM/F12 ( containing 15% fetal bovine serum, endothelial cell growth supplements (ECGs) 100 g/L, 1 ×105 units/L of penicillin and streptomycin, respectively).EPCs were characterized as double positive for DiI-Ac-LDL uptake and lectin binding.Abdominal aorta was obtained from 1 to 2 month old Sprague-Dawley rats.Vascular SMCs were cultured by tissue explant method and identified byα-SM-actin immunofluorescence.In transwell co-culture system, the confluent EPCs located in the upper chamber and SMCs were seeded on the lower cham-ber.The experiments were divided into passage 3 SMCs group (P3), passage 4 SMCs group (P4), passage 4 SMCs co-culture with EPCs derived from young rats group (P4YE) and passage 4 SMCs co-culture with EPCs derived from aged rats group (P4AE).The protein expression ofα-SM-actin and osteopontin was detected by Western blotting.[3H]-TdR incor-poration assay was used to determine the proliferation.SMC migration was analyzed by scratch wound healing assay.RE-SULTS:Compared with P3 group,α-SM-actin expression in P4 group significantly decreased and osteopontin protein ex-pression obviously increased, whereas no significant change was found in P4YE group.Compared with P4 group, confluent EPCs derived from young and aged rats both markedly increased α-SM-actin and decreased osteopontin expression in P4 SMCs.Compared with aged rat-derived EPCs, young rat-derived EPCs were more effectively to induce a delayed SMC phe-notype transition (from contractile phenotype to a synthetic phenotype), and to inhibit SMC proliferation and migration. CONCLUSION:Co-culture of confluent EPC induces a delayed vascular SMC phenotype transition and inhibits SMCs pro-liferation and migration.Young rat derived EPCs are more effective to induce a delayed vascular SMC phenotype transition and has stronger inhibitory effects on SMCs proliferation and migration compared with that derived from aged rats.

BACKGROUND:Umbilical cord as childbirth waste has wide variety of sources and can be easily obtained, without any ethical and legal restrictions. Therefore, human umbilical cord mesenchymal stem cells break al the restrictions originated from other sources of mesenchymal stem cells. OBJECTIVE:To review the application of human umbilical cord mesenchymal stem cells in cartilage diseases, neuroglioma, ischemic brain injury, lung disease, liver disease and models of myocardial infarction. METHODS:The PubMed and Wanfang databases were searched by the first author using the keywords of“human umbilical cord mesenchymal stem cells, disease models, celltherapy”in English and Chinese, respectively. Seventy-three articles were searched and final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells have multilineage differentiation capacity similar to bone marrow mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, human umbilical cord mesenchymal stem cells have lower immunogenicity. Human umbilical cord mesenchymal stem cells show certain curative effects on cartilage disease, neuroglioma, ischemic brain injury, lung disease, liver disease and myocardial infarction, indicating that human umbilical cord mesenchymal stem cells can be used for celltransplantation to treat various diseases.

Objective To understand the molecular aberration at whole genomic level,CGH (comparative genomic hybridization) was used to investigate genetic abnormality in lung cancer.Methods Comparative genomic hybridization was performed in 17 cases to detect the global genomic aberration in cancer tissue cells.Results All of 17 cases detected by CGH showed chromosomal aberrations.The average numbers of chromosomal gains and losses in each case were 7.0 and 4.8 in NSCLC and 8.4 and 9.6 in SCLC,respectively.The frequency of gains and losses on chromosome had no significant differences between NSCLC and SCLC.The frequencies of gains on chromosomal arms 3q24 -28 and 11q13(58.3％ and 58.3％ ) in NSCLC were significantly higher than that in SCLC(0％ and 0％ ) ( P ＜0.05 and ＜0.05,respectively).Conclusions The cytogenetic aberration generally existed in lung cancer cells.Several regions ( more than one) of chromosomal aberration were involved in the carcinogenesis of NSCLC and SCLC.The regions and frequencies of chromosomal aberration in NSCLC were somewhat different from that in SCLC,which might result in the different biological behavior of the two types of lung cancer.The chromosomal aberration might be served as a marker to differentiate the two types of lung cancer.

BACKGROUND:Systemic lupus erythematosus is an autoimmune disease characterized as an emergence of a variety of autoantibodies in serum and multi-system and multi-organ lesions. Currently, there is a lack of effective treatment options, and umbilical cord mesenchymal stem cel s are a promising therapy for systemic lupus erythematosus based on cel biological roles. OBJECTIVE:To observe the therapeutic efficacy of human umbilical cord mesenchymal stem cel transplantation in the treatment of systemic lupus erythematosus in mice. METHODS:Human umbilical cord mesenchymal stem cel s were isolated and cultured fol owed by labeling with DiR fluorescence. Experimental mice were divided into normal control group (C57BL mice), model control group (C57BL/lpr mice), low-, medium-and high-dose umbilical cord mesenchymal stem cel therapy groups (C57BL/lpr mice), with 10 mice in each group. Mice in the low-, medium-and high-dose groups were respectively injected 0.5×106, 1×106, 2×106 human umbilical cord mesenchymal stem cel s, once a week, for 3 consecutive weeks. At the end of treatment, blood samples were col ected to measure antinuclear antibody, anti-histone antibody, anti-double stranded DNA antibody changes;OPG and Foxp3 gene expression changes were detected by quantitative PCR method. RESULTS AND CONCLUSION:After treatment, the levels of anti-nuclear antibodies, anti-histone antibodies and anti-double stranded DNA antibodies in the peripheral blood of mice were al declined in the low-, medium-and high-dose groups, while the number of peripheral blood CD4+CD25+T cel s was significantly elevated. OPG and Foxp3 gene expression was also increased dramatical y in the low-, medium-and high-dose groups, which was similar to that in the normal control group and significantly different from that in the model control group (P<0.01). Experimental findings demonstrate that after transplantation of human umbilical cord mesenchymal stem cel s, al relevant indicators in C57BL/lpr mice recovered to the normal levels, and the high-dose treatment group had the most obvious effect.

BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.

OBJECTIVETo evaluate fluorescence in situ hybridization (FISH) in the detection of inv (16) (p13; q22).

METHODSSpectrum red labeled yeast artificial chromosome (YAC) clone 854E2 which spans the breakpoint cluster region in MYH11 in band 16p13 and single color interphase FISH were used to detect inv (16) in 26 cases of acute myelomonocytic leukemias (AML-M(4)), and the results were compared with that of conventional cytogenetic analysis.

RESULTSR banding karyotyping test revealed no inv (16) in 25 cases, one AML M(4Eo) case showed inv (16) by G banding. Nine cases including all three M(4Eo) had inv (16) by FISH analysis, among whom the characteristic fluorescence signal pattern of the inv (16) was seen in 13.3% to 32.1% (median, 21.3%) of the tested cells.

CONCLUSIONYAC 854E2 and interphase FISH provide a powerful technique in the detection of inv (16) (p13q22).

Adult ; Aged ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Interphase ; genetics ; Leukemia, Myelomonocytic, Acute ; genetics ; pathology ; Male ; Middle Aged