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Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies.
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Animals , Amino Acid Sequence , Basic Helix-Loop-Helix Transcription Factors , Genetics , Cell Differentiation , Genetics , Gene Expression Regulation, Developmental , Mesoderm , Embryology , Metabolism , Mice, Knockout , Molecular Sequence Data , Pluripotent Stem Cells , Metabolism , Sequence Homology, Amino AcidABSTRACT
Objective To investigate the application value of the self-rated health measurement scale (SRHMS) in health check-up.Method A total of 8 083 health check-up receivers frow Fujian Province underwent SRHMS self-assessment,and the results of the assessment were analyzed by Chi-square test.Result The results of the survey showed that the total prevalence rate of chronic disease was 57.8％ top five were stomach/duodenum diseases (14.57％),hypertension (11.58％),mammary gland disease (8.04％),abnormal blood lipid (7.16％) and prostate disease (6.54％).The proportion of these five kinds of illness was 43.38％.A positive correlation between the prevalence rate of chronic disease and age was found.The difference of the prevalence rate of chronic diseases between each age group was statistically significant (x2=863.816,P=0.000).Those with family history of chronic diseases accounted for 69.27％ and those with history of present illness accounted for 57.83％ (4 674/8 083).Bodily pain,dietary preferences,refined grain intake,lack of exercise,and alcohol drinking was found in 75.03％,60.15％,54.84％,46.07％ and 41.50％ participants.Conclusion The SRHMS could reflect the chronic disease incidence and unhealthy daily habits.
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Objective To investigate the effect of different concentrations of methotrexate (MTX) on IL-17 from peripheral blood mononuclear cells(PBMCs) and To clarify the active mechanisms of MTX on RA. Methods PBMCs were isolated from heparinized blood of healthy donors or patients with RA using Ficoll-Hypaque density gradient centrifugation. The cells were pretreated with various concentrations of MTX and then stimulated by anti-human CD3/anti-human CD28 at 37℃5%CO2. The IL-17 mRAN level was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The supernatants were harvested and the protein level of IL-17 was tested by ELISA kit. The percentage of CD4+IL-17+cells in PBMCs was detected by flow cytometry. Results For the four different concentrations of MTX groups (0.1,1.0, 5, 25μg/ml), the IL-17 mRNA/GAPDH ratio(0.58±0.09,0.48±0.11, 0.50±0.09, 0.51±0.14) were lower than those of the non-drug group(0.76±0.08). Paired-t test or independent-samplet test showed significant difference between the MTX treatment group and the non-drug group(P＜0.01). The level of IL-17 of the four MTX groups was(121±54)pg/ml and(104±45)pg/ml and(90±36)pg/ml and(115±41)pg/ml, which was lower than the non-drug group(370±187)pg/ml(P＜0.01). The average C D4+IL-17+cell ratio was reduced, but had no statistically signficant differences(P＞0.05). Conclusion MTX can decrease Th17 cells differentiation and suppress IL-17 production of PBMCs, but no association can be found between its effect on the expression of IL-17 and the concentration of MTX.
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Objective To detect steroidogenic acute regulatory protein (StAR) expression in apolipoprotein E-deficient mice at different ages and serum lipid levels. Methods Nighty-six C57BL/6J and apoE-/- mice were enrolled, which were divided into 16 groups with 6 mice per group according to age (1 day, 1, 3, 5 months), sex and genotype (C57BL/6J and apoE-/-). The serum lipid levels in C57BL/6J and apoE-/- mice were detected by commercial kits. StAR mRNA and protein expressions in liver were detected by Real-time PCR and Western blot respectively. Results ApoE-/- mice had higher LDL-cholesterol and lower HDL-cholesterol compared with C57BL/6J mice of the same age and sex. StAR mRNA and protein expressions were decreased following with aging in C57BL/6J mice. However, in apoE-/- mice with higher lipid levels, StAR mRNA and protein expressions were changed with the lipid levels other than ages. StAR mRNA and protein increased in the early stage, and then decreased with the increasement of lipids levels. Conclusions StAR could affect lipids levels and may be an effective regulator for atherosclerosis and other cardiovascular diseases.
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In the report of the definition and classification subcommittee of the International Dry Eye WorkShop (2007), inflammation was emphasized specially, and anti-inflammation therapy was also included in the treatment recommendations by severity level. Inflammation has been highly valued in the pathogenesis of dry eye recently. The management of dry eye through anti-inflammation therapy is also a hot point in recent years. What is the role of inflammation in the development of dry eye? This article mainly reviews the advancement of the study about their relevance in recent years.
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Objective To explore the content and gene expression of endothelial nitric-oxide synthase (eNOS) in heart of hyperlipidemia apoE-/- mice. Methods apoE-/- mice were fed with high fat diet as high-fat diet group (n= 7) and normal fat diet as control group (n = 6) for 12 weeks. Serum lipid profiles including total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) were detected, eNOS mRNA expression in heart were determined by Real-Time RT-PCR, as well as protein expression by immunohistochemistry. Results Levels of serum TC, TG and LDL clearly increased in high fat diet group, while HDL decreased (P< 0. 01). mRNA and protein expression of eNOS in heart reduced in high fat diet group(P<0.01). Conclusions High fat diet may lead to hyperlipidemia in apoE-/- mice, which may down-regulate eNOS expression and decrease eNOS content in heart.
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Objective By using percutaneous endovascular microcatheter technique to establish an animal model of coronary microembolization in mini-swine which is suitable for long-term observation.Methods Coronary microembolization was established in 10 mini-swine by infusing 15 × 10~4 microspheres (φ45μm) selectively into the left anterior descending artery (n = 10). Coronary flow reserve (CFR) was measured by Doppler wire and left ventricular eject fraction (EF) was assessed by echocardiography.Hematoxylin and eosin (HE) staining and nitroblue tetrazolium (NBT) dye were used to demonstrate the presence of microembolization after the procedure of coronary microembolization. The ultra-structures of cardiomyocyte were observed by transmission electron microscopy (TEM). Before sacrifice, the CFR measurement and coronary angiography were performed again in survival animals. Results The coronary microvascular integrity (CFR < 2.0) and left ventricular function (EF < 50% ) were damaged by coronary microembolization. One month after the procedure, all the 10 animals survived and were able to receive the angiography and CFR measurement again. HE staining and NBT dye could demonstrate the presence of microembolization. The edema and fibrosis of cardiomyocytes could be revealed with TEM. Conclusion The animal model of coronary microembolization can be established in mini-swine by using percutaneous endovascular microcatheter technique. The model is suitable for long-term observation, the preparation is technically-simple and minimally-invasive with very low mortality. Therefore, this kind of animal model is an ideal experimental form for studying the mechanism of coronary microembolization.
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<p><b>OBJECTIVE</b>To investigate the antitumor and anti-metastatic effect of in situ transduction of adenovirus encoding cytosine deaminase (AdCD) followed by the systemic use of 5-fluorocytosine (5-FC) in the orthotopic (o.t.) prostate cancer mouse model.</p><p><b>METHODS</b>The o.t. prostate cancer model of C57BL/6 mouse was developed by o.t. inoculation of RM-1 cells to the subcapsular area of the prostate gland. In situ transduction of the CD gene, followed by systemic use of 5-FC at a daily dosage of 300 mg/kg for 14 days, was performed two days later.</p><p><b>RESULTS</b>Compared with mice treated with Adbeta-gal/5-FC, 5-FC and PBS, mice of the o.t. model receiving in situ treatment of AdCD/5-FC had significant prolongation of survival and suppression of local tumor growth. More importantly, pathological observations showed that metastatic activity occurred in all mice of the PBS, 5-FC and Adbeta-gal groups including metastasis to the iliac lymph node (10/10, 10/10, 10/10) and the lung (8/10, 7/10, 7/10). However, only two out of ten had iliac lymphatic metastasis in the AdCD/5-FC group with no systemic or preaotic lymphatic metastasis, suggesting a strong metastatic inhibitory effect.</p><p><b>CONCLUSIONS</b>In situ transduction of AdCD followed by systemic use of 5-FC leads to the inhibitory effect on tumor growth and metastatic activity in the o.t. mouse model of prostate cancer. Clinically, it may be possible to treat metastatic or recurrent prostate cancer with a novel gene therapy using in situ injection techniques in future.</p>
Subject(s)
Animals , Male , Mice , Adenoviridae , Genetics , Cell Division , Genetics , Cytosine Deaminase , Disease Models, Animal , Flucytosine , Pharmacology , Lymphatic Metastasis , Genetics , Pathology , Mice, Inbred C57BL , Neoplasm Transplantation , Nucleoside Deaminases , Genetics , Prostatic Neoplasms , Genetics , Mortality , Survival Rate , Transfection , Tumor Cells, CulturedABSTRACT
Objective To study the inhibitory effect of CD/5 FC system on the prostate cancer cell DU145,RM 1 in vitro. Methods In vitro prostate cancer cells DU145 and RM 1 were infected by recombinant adenoviral vector and then the inhibitory effect and the bystander effect of CD/5 FC system were observed. Results CD/5 FC system had a remarkable inhibitory effect on prostate cancer cell DU145 and RM 1 in vitro, the inhibition rates being higher than 97% when infected by AdCMVCD after 4 days added with 15.5mmol/L 5 FC. A strong bystander effect could also be observed, more than 75% tumor cells being killed when only 10% were infected. Conclusions CD/5 FC system may serve as an effective gene therapeutic method.The existence of bystander effect makes itself available in the use of tumor treatment in vivo.
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AIM: To study the effects of VEGF over-expressed C6 glioma cells on the expression of Flk-1 and Flt-1 in cocultured microvascular endothelial cells using the rat hepatic cells BRL 3A as control. METHODS: Cocultured systems of rat pulmonary microvascular endothelial cells with C6 and endothelial cells with BRL 3A were established. Immunocytochemical method was used to investigate the expression change of Flk-1 and Flt-1 protein in cocultured microvascular endothelial cells. The expression of Flt-1 and Flk-1 mRNA was analyzed by RT-PCR and Northern blot. RESULTS: The microvascular endothelial cells cocultured with C6 showed increased expression of Flk-1 and Flt-1 protein ( P
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AIM: To study the role of c-myc oncogene in L6565 leukemia oncogenesis and the effects of therapy by inhibition of its expression with antisense c-myc. METHODS: A recombinant retroviral vector containing antisense c-myc of the murine (pGNCas)was constructed and then transfected into PA317 cells by the method of calcium phosphate precipitation. L6565 clone cells were infected with retrovirus particles. Stable integretion of antisense c-myc was shown by PCR. The change of the malignance and phenotype of L6565as were detected by the examination of the growth, morphology, cells cycle, agar assay and expression of c-myc. RESULTS: The shape of most L6565as cells became spherical. The growth of L6565as was inhibited compared to control cells. The analysis of cells cycle: L6565as cells were arrest in G 0/G 1 phase, decreased in S phase. The ability of L6565as cells to form colony in soft agarose was significantly suppressed. c-myc in L6565as cells was lowly expressed. CONCLUSION: (1)c-myc plays a critical role in L6565 leukemia oncogenesis; (2)Inhibition of expression of c-myc makes partly reversion of malignant phenotype of L6565 murine leukemia clone cells.
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A Review] The biological characteristics of viral L6565 leukemia cell clone were as follows: (1) The chromosome counts varied 38～114 , and stem cells were 42; (2) Virus particles type A and type C found in the cytoplasm of clone cells; (3) X-C assays were positive, c- myc and c- fos gene overexpressed in clone cells; (4) Differential markers CD4, CD8, CD45R were negative, CD45RO ? were positive; (5) The supernatant of clone cells could induce T or B lymphocytic leukemia/lymphoma and granulocytic leukemia in SSB strain mice. The leukemogenic effect of concentrate supernatant was stronger than non-concentrate supernatant( P
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AIM: To study the role of protein kinase B (PKB) in tryptase-induced gene expression on ECV304 cells. METHODS: The expression of PKB, transcript factor AP-1 and NF-?B P65, IL-8, JNK, p38MAPK, and the activity of PKB were measured using RT-PCR and Western blotting. RESULTS: Tryptase at concentration of 1 ?g/L increased the activity of PKB by promoting PKB phosphorylation, promoted the expression of PKB, chemokine IL-8, transcription factor AP-1 and NF-?B P65, however, no changes of JNK and p38MAPK was observed. PI3K specific inhibitor (LY294002) abolished the augment of PKB, NF-?B P65 and IL-8 expression. Antisense PKB cDNA transfection also abolished the augment of PKB, AP-1, NF-?B P65 and IL-8 expression. Though PAR2 antibody did not inhibit PKB expression, it did inhibit the phosphorylation by tryptase in ECV304 cells. CONCLUSION: These results indicate that tryptase can activate PKB through PAR2 receptor and subsequently NF-?B, AP1, IL-8 and PKB expression.
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AIM: To study the effect of hypoxia on CD73 expression in mouse microvascular endothelial cell line bEnd.3. METHODS: ① bEnd.3 cells were exposed to different periods of hypoxia. ② Concentration of LDH released by bEnd.3 cells into the culture medium was detected. ③ Surface CD73 activity in bEnd.3 cells was measured by HPLC according to the conversion of E-AMP to E-ADO. ④ CD73 mRNA expression were analyzed by semiquantitative RT-PCR. ⑤ Cell surface proteins were biotinylated and CD73 was detected by avidin blots of immunoprecipitation with mAb TY23. RESULTS: ① bEnd.3 cells exposed to hypoxia for 24 h demonstrated a significant increase in LDH release (P
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AIM: To study the cytological characteristics and gene expression of normal cultured bEnd.3, a mouse brain microvascular endothelial cell strain. METHODS: The morphology of bEnd.3 was studied by light and electronic microscopy, its molecular markers were observed by immunocytochemistry. Cell proliferation kinetics and apoptosis were analyzed by flow cytometry and MTT assay, PGE_2 level was measured by ELISA, and expression of the genes that closely related with vascular endothelial functions was studied by gene micro-array. RESULTS: bEnd.3 had morphological characteristics of microvascular endothelial cells (MVEC) growing in a cobblestone pattern, forming tube-like structure or capillary network and having microvilli. Furthermore, bEnd.3 showed positive staining for vW factor and CD34 and secreted high level of PGE_2 (644.55?30.24 ng/L). Gene micro-array analysis showed CD31, CD36, CD105 expression, and other genes closely related to microvascular endothelial functions also expressed at relatively high level. In addition, bEnd.3 responsed sensitively to mitogen such as basic fibroblast growth factor. CONCLUSION: bEnd.3 is a kind of MVEC, and it can be utilized to study the mechanisms of some diseases such as cancers and cardio- cerebral vascular diseases.
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AIM: To isolate, purify and differentiate endothelial progenitor cells from cord blood in vitro and to study their biological characteristics. METHODS: CD133~+ cells were selected from fresh cord blood mononuclear cells (MNC) by magnetic activated cell-sorting system (MACS). EPC was studied by flow cytometry, immunocytochemistry and immunofluorescence staining. Isolated cells were cultured in IMDM medium supplemented with or without VEGF, bFGF, SCF. RESULTS: The percentage of CD133~+ cells of cord blood MNC was (1.41?1.14)%, and purity was 75%-85% (FACS method). CD133~+ cells were grown on fibronectin-coated chamber slides in the presence of VEGF, bFGF, SCF. Within 1-2 hours of culture cells became adherent. On day 7-10, the adherent cells displayed a typical "cobblestone" morphology. After 14 days of culture, the adherent cells revealed a heterogeneous cell population, comprising small-sized round cells, spindle-like cells and formed tube-like structure. Weibel-Palade bodies were shown on the transmission electron microscopy photomicrographs. Compared with the (original,) cell markers CD133 and CD34 decreased significantly (77.0%?3.3% to 1.6%?2.2% and 93.1%?4.7% to 37.4%?4.9%, P
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It was found that protooncogene c-fos was over-expressing in SAC-IIB_2 cell strains. Targeting the code 2-7 sequence of c-fos cDNA of mice, we designed and synthesised eighteen mers antisense. sense and nonsense oligodeoxynucleotides(ODN). The results indicated that antisense c-fos ODN could induce SAC- IIB_2 maturative differentiation, inhibit its growth rate, decrease its ability of forming tumor in nude mice and expression of Fos protein, however, antisense ODN has no remarkable effect on cellular dynamics of SAC-IIB2. But sense and nonsense c-fos ODN have no similar effect on SAC-IIB_2 cell strains. The results demonstrated that antisense c-fos oligodeoxynucleotide has inhibition effects on malignant phenotype of mice SRS lymphoma cell.
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Thyptase,a predominant serine protease in mast cells,is released by mast cell degranulation in an enzymatically active form.Recent research suggests that ttyptase plays important roles in mast cell mediated anaphylaxis,angiogenesis,initiation and promotion of neoplasm.This paper reviews tryptase's struture,molecular biology,biochemistry,processing,activation and its relationship with diseases.
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Cytosine deaminase (CD) gene is a kind of suicide gene, cytosine deaminase can catalyze the conversion from 5 - fluorocytosine (5 - FC) into 5 - fluorouracil (5 - FU), this combination therapy (CDgene/5 - FC) were widely used in cancer therapy. The results of many researches in different cancer cells have demonstrated that it can inhibit tumor growth significantly. The mechanism includes 5 - FU's direct killing tumor cells and its bystander effect. It can also be used as a potential purging method for autologous hematopoietic cells transplantion after chemotherapy in breast cancer patients. CD/5 - FC combined with other therapies would be an important future research approach.
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The growth and spread of malignant neoplasms largely depend on angiogenesis.Recent studies demonstrate that a various types of neovascularization including angiogenesis,vasculogenesis and vasculogenic mimicry exist in a few highly aggressive tumors.Ovarian carcinoma is the leading cause of death in gynecologic malignancy.Here,we review the effect of angiogenesis,vasculogenesis and vasculogenic mimicry in development and spread of ovarian carcinoma.