The role of Notch signaling pathway in immune regulation of sepsis / 国际儿科学杂志

Sepsis is a clinical syndrome manifested by organ dysfunction due to disordered inflammatory response after severe infection.The occurrence, development and prognosis of sepsis are closely related to the immune regulation of the body.The essence of sepsis is that the state of excessive proinflammatory response in the early stage gradually progresses to the state of immunosuppression in the late stage, which leads to the body′s inability to resist inflammation and endangers life.As a highly conserved signaling pathway, Notch pathway has the ability to regulate cell growth and differentiation, and participates in the occurrence and development of various inflammatory diseases.In recent years, the important role of Notch signaling pathway in the occurrence and development of sepsis has attracted extensive attention.This article mainly reviews the role of Notch signaling pathway in immune regulation of sepsis.

Effect of Modified Shengjiangsan on Notch Signal Pathway and Podocyte Apoptosis in MN Rats / 中国实验方剂学杂志

Objective:The purpose of this article was to observe the effect of modified Shengjiangsan on podocyte apoptosis in membranous nephropathy （MN） rats， to explore the molecular mechanism of its treatment of MN and to provide experimental basis for its clinical application. Method:The MN rat model was established by injection of cationic bovine serum albumin into the tail vein of rats. The successfully modeled rats were then randomly divided into model group （equal volume of normal saline）， modified Shengjiangsan group （27.3 g·kg^-1） and benazepril group （10 mg·kg^-1）， with corresponding drug dosage once a day for 4 weeks of continuous intervention. After drug administration， the 24-hour urine protein （UTP） was detected. Real time fluorescent quantitative polymerase chain reaction （Real-time PCR） and immunohistochemical （IHC） methods were used to detect Podocalyxin， Nephrin， Podocin， Synaptopodin mRNA and protein expression levels in rat kidney tissue. terninal-deoxynucleoitidyl transferase medsated nick and labeling （TUNEL） method was used to detect cell apoptosis rate in rat kidney tissue， and Western blot was used to detect Notch1， Hes1， B lymphoblastoma-2 （Bcl-2） associated X protein （Bax）， and Bcl-2 protein expression levels in rat kidney tissue. Result:Compared with the normal group， UTP in the model group increased significantly， renal tissue cell apoptosis increased significantly， podocyte marker proteins podocalyxin， Nephrin， Podocin， Synaptopodin mRNA and protein expression levels decreased significantly， and Notch1， Hes1， Bax protein expression increased significantly， and Bcl-2 protein expression was significantly reduced（<italic>P</italic><0.05）. Compared with the model group， UTP levels in MN rats were significantly reduced in modified Shengjiangsan and benazepril groups， with reduced rate of renal cell apoptosis， increased mRNA and protein expression levels of podocalyxin， Nephrin， Podocin， and Synaptopodin in renal tissue， decreased Notch1， Hes1， Bax protein expression， and increased Bcl-2 protein expression（<italic>P</italic><0.05）. Conclusion:Modified Shengjiangsan can inhibit the Notch signaling pathway， reduce the apoptosis of rat kidney tissue podocytes， and reduce the kidney injury of MN rats.

Inhibitory Effect of Essential Oil from Fructus Alpinia zerumbet on Endothelial-to-mesenchymal Transition Induced by High Glucose via Nrf2/Notch1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the inhibitory effect and the possible mechanism of essential oil from fructus Alpinia zerumbet （EOFAZ） on endothelial-to-mesenchymal transition （EndMT） induced by high glucose （HG）. Method:Human umbilical vein endothelial cells （HUVECs） was cultured in vitro to analyze the pharmacodynamic effects of EOFAZ on EndMT and oxidative stress damage induced by HG. The experiment was set the blank group， HG group （35 mmol·L-1）， EOFAZ low dose group （1 μg·L-1） and EOFAZ high dose group （4 μg·L-1）. After EOFAZ intervention for 2 h， HG was added to incubate for 72 h in order to establish EndMT cell model. Western blot was used to detect the protein expression of vimentin and platelet endothelial cell adhesion molecule （CD31）. Angiogenesis experiment was used to detect the ability of cell migration ability in order to analyze the effect of EOFAZ on EndMT. The changes of reactive oxygen species （ROS） levels were detected by 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescence probe and the contents of malondialdehyde （MDA）， superoxide dismutase （SOD） and catalase （CAT） in cells were detected by the kit method to analyze the effect of EOFAZ on oxidative stress. Western blot was used to detect the protein expression levels of nuclear transcription factor E2 related factor 2 （Nrf2） and Notch1. The overexpression of Nrf2 was achieved by adenovirus （AD） transfection and the mechanism of EOFAZ inhibiting EndMT was further analyzed. The experiment was set the blank group， HG group （35 mmol·L-1）， AD-Nrf2 group， EOFAZ group （4 μg·L-1）， AD-Nrf2+EOFAZ group （4 μg·L-1）. The cells were infected with recombinant adenovirus overexpression plasmid of Nrf2 gene for 6 h， then replaced with normal medium for 24 h. After EOFAZ intervention for 2 h， HG was added to co-incubate for 72 h to induce EndMT. Western blot was used to detect the protein expressions of Nrf2， CD31， vimentin， Notch1 and Snail. Result:Compared with the HG group， after treatment with EOFAZ， the protein expression of CD31 was significantly up-regulated （P<0.05）， the protein expression of vimentin was significantly down-regulated （P<0.01）， the ability of cell migration was decreased （P<0.01）， and the contents of ROS and MDA were decreased （P<0.05， P<0.01）， the levels of CAT and SOD were increased （P<0.01）. In addition， EOFAZ could significantly up-regulate the protein expression of antioxidant signal Nrf2 （P<0.01） and down-regulate the protein expression of Notch1 （P<0.01）. High expression of Nrf2 was achieved by stable AD transfection into HUVECs. The results of Western blot showed that， compared with the HG group， the protein expression levels of Nrf2 and CD31 in each treatment group were significantly increased （P<0.01）， while the protein expression levels of vimentin， Notch1 and Snail were down-regulated （P<0.01）. At the same time， compared with the AD-Nrf2 group， the AD-Nrf2+EOFAZ group could further up-regulate the protein expressions of Nrf2 and CD31 （P<0.05， P<0.01）， while decrease the protein expression levels of vimentin， Notch1 and Snail （P<0.01）. Conclusion:EOFAZ ameliorates oxidative stress injury of vascular endothelial cells induced by HG and inhibits EndMT， which is related to Nrf2/Notch1 signaling pathway.

Effects of tumor necrosis factor-α on osteogenic differentiation and Notch signaling pathway in human periodontal ligament stem cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of tumor necrosis factor-α (TNF-α) on osteogenic differentiation and Notch signaling pathway of periodontal ligament stem cells (PDLSCs) and to investigate the regulatory role of Notch signaling pathway on the osteogenic differentiation of PDLSCs under the influence of TNF-α.</p><p><b>METHODS</b>PDLSCs were obtained through enzyme digestion and tissue block method. The expression levels of stem cell surface markers CD105, CD90, CD146, CD45, and CD31 were detected by fluorescence activated cell sorter (FACS). PDLSCs were divided into experimental (10 ng·mL⁻¹ TNF-α) and control groups (0 ng·mL⁻¹ TNF-α). The proliferation ability of PDLSCs was detected using cell counting kit-8 (CCK-8). The effect of TNF-α on the osteogenic ability of PDLSCs were tested by measuring alkaline phosphatase (ALP) activity and conducting alizarin red staining and quantitative real-time polymerase chain reaction (PCR). We tested Notch signal pathway receptors Notch1, Notch2, ligand JAG1, JGA2, and downstream gene Hes-1. Changes in DLL1 expression were detected by quantitative real-time PCR.</p><p><b>RESULTS</b>FACS profiling showed that PDLSCs were strongly positive for CD105, CD90, and CD146 but negative for CD45 and CD31. CCK-8 results showed that TNF-α could promote the proliferation of PDLSCs (P<0.05). ALP activity in the experimental group was lower than that in the control group (P<0.05). Alizarin red staining showed that the experimental group had decreased mineralized nodules as compared with the control group. Quantitative real-time PCR results showed that the mRNA expression of osteogenic marker genes cementum attachment protein (CAP), osteopontin (OPN), and Runt-related transcription factor 2 (Runx2) significantly decreased in the experimental group as compared with those in the control group (P<0.05). The expression levels of Notch1, Notch2, JAG1, JGA2 and Hes-1 were significantly decreased (P<0.05), whereas those of Notch3 and DLL1 were increased in Notch signaling pathway-related molecules (P<0.05).</p><p><b>CONCLUSIONS</b>TNF-α can promote PDLSCs proliferation and inhibit bone differentiation and Notch signaling pathway expression, indicating that the Notch signaling pathway regulates PDLSCs osteogenic differentiation.</p>

Study on davallia mariesil flavones improving condition of osteoporosis via Notch signaling pathway / 中国免疫学杂志

Objective:To evaluate the effect and mechanism of davallia mariesil flavones (DMF) improving osteoporosis via Notch signaling pathway.Methods:(1) 120 cases patients with osteoporosis in our hospital from January 2016 to January 2017 were analyzed,and divided into experiment group (60 cases) and control group (60 cases) using digital random grouping method.Experiment group was treated with DMF combined with D-calfor.Control group was treated with D-calfor only.After treatment, the levels of serum calcium,serum phosphor,TNF-α,MCP-1 and IL-6 in serum were detected to evaluate the clinical efficacy.(2) Bone marrow stromal cells were separated and cultivate.NC group:DMEM(contain 10% FBS).RA group:RA(0.4 mmol/L).DMF+RA group:DMEM(contain DMF)+RA(0.4 mmol/L).Jaggedl+RA group:Jaggedl(1 000 μg/L)+RA(0.4 mmol/L).Jaggedl+DMF+RA group:Jaggedl(1 000 μg/L)+DMEM(contain DMF)+RA(0.4 mmol/L).DAPT+RA group:DAPT(16 μmol/L)+RA(0.4 mmol/L). DAPT+DMF+RA group:DAPT(16 μmol/L)+DMEM(contain DMF)+RA(0.4 mmol/L).Western blotting assays and PCR were performed to assess mRNA and protein levels of Notch-1,Hes-1.Results: (1) In clinical study,the effective rate in treatment group was obviously higher than control group (91.67%>76.67%,P<0.05).The levels of serum calcium and serum phosphor in the experiment group was higher than in the control group (P<0.05).The levels of TNF-α,MCP-1 and IL-6 in the experiment group was lower than in the control group (P<0.05).(2) In experimental study,compared with the RA group,the expressions of Notch-1,Hes-1 mRNA and protein were upregulated in Jaggedl+RA group,but were downregulated in DAPT+RA group,DMF+RA group (P<0.05). Compared with the Jaggedl+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in Jaggedl+DMF+RA group (P<0.05).Compared with the DAPT+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in DAPT+DMF+RA group (P<0.05).Conclusion:DMF could improve the condition of osteoporosis.The mechanism may be associated with inhibiting the Notch signaling pathway by DMF.

DAPT regulates Treg/Th17 cells immune balance to inhibit atherosclerosis / 中国免疫学杂志

Objective:To observe the effect of Notch signal inhibitor DAPT (γ-secretase inhibitor ) on the pathological changes of atherosclerosis mice and the immune balance of Treg/Th17.Methods:24 ApoE knockout C57BL mice were randomly divided into blank group,model group and DAPT group.The blank group were fed with normal diet ,the model group and the experimental group were fed with high fat diet.After 5 weeks of feeding,the mice in the experimental group were injected with DAPT [100 mg/(kg· d),re-suspended in DMSO],and the other two groups were injected with the equivalent amount of DMSO .After another 5 weeks,pathological changes of the mice in each group were analyzed by HE staining .ELISA was used to detect the level of IL-17 in plasma,and the propor-tion of splenic Treg/Th17 cells in each group was detected by flow cytometry .Results:HE staining results showed that the model group had obvious plaque formation and foam cell formation ,which showed that the AS model was successfully prepared .The degree of arterial disease in the DAPT group was significantly less than that in the model group .The plasma levels of IL-17 in the blank group , model group and DAPT group were(293.94±28.59),(454.05±172.68) and (335.40±89.57) pg/ml,respectively .The percentages of Treg cells in the blank group,model group and DAPT group were(3.80±0.56)%,(2.54±0.38)%and(4.73±0.64)%,respective-ly.The Th17 cell subsets of mice in the blank group , model group and DAPT group were ( 3.46 ±0.23 )%, ( 4.52 ±0.85 )% and (1.38±0.37)%,respectively .Conclusion:DAPT decreased the plasma level of IL-17 in AS mice,inhibited the differentiation of Th17 cell subsets,and promoted the differentiation of Treg ,and reduced the atherosclerosis by changing the Treg/Th17 cells immune balance.

Progress on the cell-surface markers and signaling pathways of colorectal cancer stem cells / 基础医学与临床

Colorectal stem cells have many bio-markers, including Lgr5 which expression is associated with THE stage of disease , also regulating the cell cycle , anothers is +4 stem cell , which is associated with tumor heteroge-neity, also expressed Bmi1, arresting cell cycle.Besides there is Msi1.Many studies show that those markers are highly expressed in colorectal cancer , which activate Notch and Wnt signaling pathway , and can promote the pro-gress of tumor .

Inhibitory effect and mechanism of epigallocatechin-3-gaUate on autogenous vein graft stenosis in rat models / 中国胸心血管外科临床杂志

@#Objective    To investigate the effect and mechanism of epigallocatechin-3-gallate (EGCG) on restenosis of the vein graft. Methods    Totally 90 Sprague-Dawley rats were randomly divided a the control group, a vein graft group and an EGCG+vein graft group. At week 1, 2 and 4, the intimal and tunica thickness of the venous graft wall was evaluated by hematoxylin-eosin staining, and the expression of Ki-67 was assessed by immunohistochemistry analysis, and then the expression of hairy and enhancer of split-1 (HES1) was measured by Western blot assay. Results    At week 2, the intimal thickness (46.76±4.89 μm vs. 8.93±0.82 μm, 46.76±4.89 μm vs. 34.24±3.57 μm), tunica thickness (47.28±4.37 vs. 16.33±1.52 μm, 47.28±4.37 vs. 36.27±3.29 μm), positive cell rate of Ki-67 (21.59%±2.29% vs. 1.12%±0.22%, 21.59%±2.29%vs. 15.38%±1.30%), expression of HES1 respectively increased in the experimental group than those in the control group and the EGCG+vein graft group (P<0.05, respectively). At week 4, the intimal thickness (66.38±6.23 μm vs. 8.29±0.79 μm,   66.38±6.23 μm vs. 48.39±4.23 μm), tunica thickness (63.27±6.18 μm vs. 15.29±1.49 μm, 63.27±6.18 μm vs. 44.63±4.49 μm), positive cell rate of Ki-67 (33.19%±3.03% vs. 1.09%±0.19%, 33.19%±3.03% vs. 24.37%±2.73%), expression of HES1 increased in the experimental group than those in the control group and EGCG+vein graft group (P<0.05, respectively). Conclusion    EGCG may inhibite restenosis of vein graft by inhibiting Notch signal pathway.

Directional differentiation of mouse mesenchymal stem cells into neuronal cells induced by salidroside via Notch and BMP signal pathway / 中草药

Objective: To study the molecule mechanism of salidroside inducing mesenchymal stem cells (MSCs) to directionally differentiate into neuronal cells via bone morphogenetic protein (BMP) and Notch signal pathways. Methods: Experiments were divided into control, induced, and blocked groups. The technologies, such as immunofluorescence, real-time PCR, and Western blotting were used to analyze the effect of salidroside on cellular proliferation, morphosis, and BMP and Notch signal pathways. Results: The immunofluorescence results showed that salidroside could affect cellular proliferation and induce MSCs to form the morphosis of neuronal cells. The positive rate of Notch1 and Jadge1 was significantly decrease to compare with the control (P < 0.05), real-time PCR results indicated that mRNA expression of Notch1 and Hes1 was obviously down-upregulated when treated with salidroside for 12-72 h (P < 0.05). However, Notch signal pathway was blocked with DAPT, a special inhibitor of Notch, the marker molecules of neuronal cells expression, such as neuron-specific enolase (NSE), microtubule associated protein 2 (MAP2), and β-tubulin III, were significantly increased when cells were treated with salisroside (P < 0.05). The mRNA levels of Smad5 and Smad8 were up-regulated when cells were treated with salidroside for 12 h, expression of Smad1/5/8 protein was increased at 12 and 24 h. When BMP signal pathway was blocked with Noggin, a special inhibitor of BMP, NSE, MAP2, and β-tubulin III mRNA expression was decreased to compare with the salidroside induced group (P < 0.05); When Notch and BMP signal pathways were simultaneously blocked with DAPT and Noggin, MAP2 and β-tubulin III mRNA expression was increased more obviously than that of the blocked with Noggin. Meanwhile the expression of NSE and β-tubulin III protein was also up-regulated (P < 0.05). Conclusion: Salidroside promotes neuron-like differentiation of MSCs by negatively regulating the Notch pathway and activating BMP signal pathway, it plays a vital role for salidroside to inhibit Notch pathway on affecting MSCs differentiation.

Effects of Osthole on Differentiation of Neural Stem Cells in vitro / 医药导报

Objective To investigate the effects of osthole on neural stem cells ( NSCs) differentiation and explore the potential mechanism. Methods Brain-derived NSCs from newborn mice were isolated and cultured in vitro and determined by immunofluorescence. The P5 generations of NSCs were placed in culture solution with osthole at concentrations of (0,10,50, 100 μmol·L-1 ) . The neuron, astrocyte and oligodendroglia cell differentiation were determined by immunofluorescence. The mRNA expression of Notch 1 and its target genes Mash 1 and Neurogenin 2 were assessed by RT-PCR. Results The neurosphere displayed Nestin and Sox 2-postive by immunofluorescence, suggesting that the cultured cells were NSCs. Osthole promoted NSCs differentiating into more neuron(P<0. 01) and oligodendrocyte(P<0. 05), but not astrocyte. Meanwhile, osthole significantly reduced the mRNA expression of Notch 1(P<0. 01) and increased Ngn 2(P<0. 01)at the dose of 100 μmol·L-1. Conclusion Osthole enhances NSCs differentiating into more neuron and oligodendrocyte via probablly inhibiting Notch signal pathway.

Notch signaling pathway regulates epithelial-mesenchymal transition and affects the invasiveness and drug resistance of bladder cancer / 基础医学与临床

Objective To investigate the effect of notch signaling pathway on drug resistance and invasiveness of bladder cancer .Methods We observed the changes of growth and morphology of bladder cancer T 24 , 5637 and J82 cells which treated for 48 hours using γ-secretase inhibitor by inverted microscope .The mRNA and protein lev-els of the EMT molecular markers , including E-cadherin , N-cadherin , vimentin and Alpha-smooth muscle actin were examined by RT-PCR and Western blot in bladder cancer cells;Detected the changes of drug resistance and invasion respectively by MTT and Transwell in bladder cancer cells .Results After completely blocking the Notch signaling pathway , the inverted microscope showed that bladder cancer cells became smaller and more disperse ;RT-PCR and Western blot showed the mRNA and protein levels of E-cadherin were up-regulated ( P<0.05 ) , contrast , N-cadherin , vimentin and Alpha-smooth muscle actin were down-regulated ( P<0.05 ); The prolifera-tion of bladder cancer cells were significantly inhibited by MTT test;The number of through microporous membrane cells significantly decreased ( P<0.05 ) shown by Transwell test .Conclusions The Notch signaling pathway is completely blocked that nhibites proliferation and EMT of bladder cancer cells , reduces drug resistance and inva-sion in bladder cancer cells .It suggests that drug resistance and invasiveness of bladder cancer can be changed through EMT which is regulated through notch signaling pathway .

Effect of spinal cord extracts after spinal cord injury on proliferation of rat embryonic neural stem cells and Notch signal pathway in vitro

OBJECTIVE@#To investigate the effect of the spinal cord extracts (SCE) after spinal cord injuries (SCIs) on the proliferation of rat embryonic neural stem cells (NSCs) and the expressions of mRNA of Notch1 as well as of Hes1 in this process in vitro.@*METHODS@#The experiment was conducted in 4 different mediums: NSCs+PBS (Group A-blank control group), NSCs+SCE with healthy SD rats (Group B-normal control group), NSCs+SCE with SD rats receiving sham-operation treatment (Group C-sham-operation group) and NSCs+ SCE with SCIs rats (Group D-paraplegic group). Proliferative abilities of 4 different groups were analyzed by MTT chromatometry after co-culture for 1, 2, 3, 4 and 5 d, respectively. The expressions of Notch1 and Hes1 mRNA were also detected with RT-PCR after co-culture for 24 and 48 h, respectively.@*RESULTS@#After co-culture for 1, 2, 3, 4 and 5 d respectively, the MTT values of group D were significantly higher than those of group A, group B and group C (P0.05). Both the expressions of Notch1 and Hes1 mRNA of group D were significantly higher than those of other 3 groups after co-culture for 24 h and 48 h as well (P0.05). There was no difference in expressions of Notch1 and Hes1 mRNA between 24 h and 48 h treatment in group D.@*CONCLUSIONS@#SCE could promote the proliferation of NSCs. It is demonstrated that the microenvironment of SCI may promote the proliferation of NSCs. Besides, SCE could increase the expression of Notch1 and Hes1 mRNA of NSC. It can be concluded that the Notch signaling pathway activation is one of the mechanisms that locally injured microenvironment contributes to the proliferation of ENSC after SCIs. This process may be performed by up-regulating the expressions of Notch1 and Hes1 gene.

The Notch signal path is involved in the protection of radiation-induced apoptosis of C17.2 neural stem cells by exogenous basic fibroblast growth factor / 中华行为医学与脑科学杂志

Objective To observe the effects of exogenous basic fibroblast growth factor (bFGF) on radiation-induced apoptosis of C17.2 neural stem cells (NSCs) with γ-secretase inhibitor (DAPT) condition and explore the relationship between bFGF and Notch signal pathway.Methods The cell viability was detected by using the MTF method.After the cells attached to the flasks,different concentrations of DAPT was added in accordance with the experimental design and cultured cells for 24 h.C17.2 NSCs were subjected to irradiation exposure by linear accelerator and treated with bFGF (40 ng/ml) 5 min after the exposure.After 48 hours,the apoptosis of cells was detected by using Flow Cytometry.Results After adding in DAPT,the cells growth was inhibited and depended on the concentrations of DAPT.Compared with the control group,all groups had statistically significant differences(P＜0.05).Flow cytometry showed compared with the control group all groups had significant differences (P＜0.05).The apoptosis rate was (11.53±0.81)％ in radiation group,(7.18±0.0.94)％ in radiation+bFGF group,(9.82±0.77) ％ in DAPT group,(21.45±0.98) ％ in Radiation+DAPT group and (10.26+ 1.03) ％ in Radiation+ DAPT +bFGF group.Between Radiation + bFGF group and Radiation group,it had statistically significant difference(P＜0.05).The pairwise comparisons of DAPT group and Radiation + DAPT group which had the same DAPT concentration had statistically significant differences (P＜0.05).The pairwise comparisons of Radiation + DAPT+bFGF group and Radiation + DAPT group which had the same DAPT concentration had statistically significant difference(P＜0.05).Conclusion Fx ogenous bFGF can inhibit apoptosis of C17.2 NSCs.Notch signaling patbway inhibitor DAPT can promote apoptosis of C17.2 NSCs which are subjected to irradiation exposure by linear accelerator and bFGF can weak apoptosis.bFGF protective effect on radiation-induced neural stem cells may be related to the Notch signaling pathway.

Construction of Notch1(NICD) Eukaryotic Expression Vector and Its Influence on the Proliferation and Differentiation of Rat Bone Marrow Mesenchymal Stem Cells in vitro / 天津医药

Objective To investigate the effect of construct the Notch1 (NICD) eukaryotic expression vector on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were experimented as the object. NICD eukaryotic expression vector was constructed. pEGFP-N1-NICD expressing plasmids were used to transfect BMSCs. The study included control group (CON group), empty vector group (VEC group) and the trans-fection group (TRA group). After 48-hour transfection, BMSCs were observed for general morphology. The protein expres-sions of NSE, GFAP and Notch1 were detected by real-time PCR and Western blotting assay respectively. The apoptosis, cy-cle distribution and cell proliferation were evaluated by flow cytometry and MTT assay. Results The DNA sequencing con-firmed that the pEGFP-N1-NICD recombinant plasmid was successfully constructed, and both VEC group and TRA group expressed green fluorescence after 48-hour transfection. The relative expression levels of Notch1 and GFAP mRNA and pro-tein were significantly higher in TRA group than those in VEC group and CON group (P<0.05), and there was no significant difference between VEC group and CON group. After 48-hour transfection, the ratio of living cells was significantly lower in TRA group than that of CON group and VEC group, and the early apoptotic rate and late apoptotic rate were significantly higher in TRA group than those of CON group and VEC group (P<0.05). The late apoptotic rate was significantly higher in VEC group than that of CON group. The proportion of G1/G0 cells was significantly higher in TRA group than that of CON group and VEC group, but S and G2/M cells were significantly lower (P<0.05). The value of growth curve was gradually de-creased in TRA group than that of CON group and VEC group (P<0.05). Conclusion The high expression of NICD gene might induce apoptosis of BMSCs, inhibit the proliferation in part, and induce into glial-like cell differentiation.

Molecular targeted therapy for breast cancer patients based on Notch signal transduction pathway / 国际肿瘤学杂志

Studies show that the Notch signal transduction pathway involves in the regulation and control of cell proliferation,differentiation and apoptosis.Aberrant Notch signaling transduction pathway can induce breast cancer in transgenic mice.High expressions of either Notch receptors or their ligands have been linked to poor prognosis in patients with breast cancer.Inhibition of Notch signal transduction pathway may be beneficial for breast cancer treatment.

Notch signal pathway and the pathogenesis of multiple myeloma / 国际肿瘤学杂志

Notch signal pathway can regulate the morphogenesis,apoptosis and cellular proliferation of normal tissues and organs,which plays an important role in regulating hematopoietic cells proliferation and differentiation in bone marrow microenvironment.The occurrence and development of multiple myeloma(MM)are closely related to the bone marrow microenvironment in which many signal pathways are involved.Recent studies show that Notch signal pathway promotes the development and progression of many cancers including MM,which plays key roles in tumor invasion and drug resistance.This review focus on the recent findings on Notch signal pathway in MM,and reveals that Notch signal pathway will be identified as a potential new therapeutic target in MM.

The mechanism underlying the differentiation of human umbilical cord-derived mesenchymal stem cells into myocardial cells induced by 5-azacytidine.

Objective: To investigate the molecular mechanism underlying the differentiation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) into myocardial cells induced by 5-azacytidine (5-aza), and to explore the expression and significance of DLL4-Notch signaling in this process. Materials and Methods: hUCMSCs were isolated and purified from the umbilical cords of normal or cesarean term deliveries under sterile conditions. After treatment with 5-aza for 24 h, hUCMSCs was continued to culture, the expression of GATA4 and NKx2.5 at 4 weeks after induction, DLL4 and Notch1 mRNA at 1d, 3d, 5d, 7d after induction were detected. The expression of cardiac troponin I (cTnI) after 4 weeks was determined by immunocytochemistry. Results: hUCMSCs treated with 5-aza were stained positively for cTnI 4 weeks after induction. The expression of Notch1 and DLL4 mRNA in the 5-aza-induced group was stable and significantly higher than that in the control group (mean Ct value for the Notch1 gene: 0.51 ± 0.21 in the 5-aza-induced group vs. 7.85 ± 0.35 in the control group; mean Ct value for the DLL4 gene: 1.60 ± 0.49 in the 5-aza-induced group vs. 12.42 ± 0.73 in the control group). Similar results were observed for Nkx2.5 and GATA4 genes. The expressions of Nkx2.5 and GATA4 mRNA in the 5-aza group were 4.72 ± 0.58 and 3.76 ± 0.06 times higher than that in the control group, respectively, with statistical significance. Conclusions: hUCMSCs can be differentiated into myocardial cells by 5-aza induction in vitro. 5-Aza may affect this process by regulating the expression of GATA4 and Nkx2.5 genes. The DLL4-Notch signal pathway may be involved in this process.

Effect of Liuwei Dihuang decoction and its decomposed recipes on gene expression related to thymocytes differentiation in normal mice and SAMP8 / 中国免疫学杂志

Objective:To research the effect of Liuwei Dihuang decoction and its decomposed recipes on gene expression related to thymocyte differentiation in normal mice and SAMP8.Methods:The expression of Notch1,Presenilin1,Presenilin2,HES1 mRNA was detected by quantitative real-time fluorescence PCR assay.Results:After oral administation of Liuwei Dihuang decoction(5,10,20 g/kg),the expressions of Presenilin2 and HES1 gene in thymocytes of normal mice were increased.After oral administation of Liuwei Dihuang decoction(10 g/kg)and SB,SX,the expressions of Presenilin2 and HES1 in thymocytes of normal mice were decreased.Conclusion:It is suggested that Liuwei Dihuang Decoction can increase the Notch signal intensity in the normal mice thymocytes and decrease that in SAMP8,which is immuno-senescenced mice.