Notch Signaling Mediates Radiation-Induced Smooth Muscle Cell Hypermuscularization and Cerebral Vasculopathy.

BACKGROUND: Emerging evidence highlighted vascular injury in aggravating radiation-induced brain injury (RIBI), a common complication of radiotherapy. This study aimed to delineate the pathological feature of cerebral small vessel and investigate the functional roles of Notch signaling in RIBI. METHODS: Brain tissue and functional MRI from RIBI patients were collected and analyzed for radiation-induced vasculopathy. A RIBI mouse model was induced by a single dose of 30-Gy cranial irradiation. Vascular morphology, pulsatility, and reactivity to pharmacological interventions, such as nimodipine and 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid, were monitored by 2-photon imaging in mice at 6 weeks postirradiation. Western blot, real-time quantitative PCR, immunofluorescence staining, and behavioral tests were performed. The effect of N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-s-phenylglycinet-butyl ester, a Notch inhibitor, was used to investigate the vascular pathogenesis of RIBI mouse model. RESULTS: Morphologically, radiation resulted in vascular malformation featured by focal contractile rings together with general stenosis. Functionally, radiation also led to hypoperfusion, attenuated vascular pulsatility, and decreased dilation to nimodipine and 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid. Mechanically, Notch activation and increased expression of α-SMA protein were found in both surgical specimens of RIBI patients and the irradiated mice. Importantly, Notch inhibition by N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-s-phenylglycinet-butyl ester significantly alleviated cerebral hypoperfusion, vasculopathy, and cognitive deficits in the RIBI mouse model. CONCLUSIONS: Radiation-induced cerebral vasculopathy showed bead-like shape and increased contractile state. Inhibition of Notch signaling by N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-s-phenylglycinet-butyl ester effectively attenuated vasculopathy and relieved cognitive impairment, suggesting Notch signaling as a therapeutic target for the treatment of RIBI.

Notch1 Has an Important Role in ß-Cell Mass Determination and Development of Diabetes.

BACKGROUND: Notch signaling pathway plays an important role in regulating pancreatic endocrine and exocrine cell fate during pancreas development. Notch signaling is also expressed in adult pancreas. There are few studies on the effect of Notch on adult pancreas. Here, we investigated the role of Notch in islet mass and glucose homeostasis in adult pancreas using Notch1 antisense transgenic (NAS). METHODS: Western blot analysis was performed for the liver of 8-week-old male NAS mice. We also conducted an intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test in 8-week-old male NAS mice and male C57BL/6 mice (control). Morphologic observation of pancreatic islet and ß-cell was conducted in two groups. Insulin secretion capacity in islets was measured by glucose-stimulated insulin secretion (GSIS) and perifusion. RESULTS: NAS mice showed higher glucose levels and lower insulin secretion in IPGTT than the control mice. There was no significant difference in insulin resistance. Total islet and ß-cell masses were decreased in NAS mice. The number of large islets (≥250 µm) decreased while that of small islets (<250 µm) increased. Reduced insulin secretion was observed in GSIS and perifusion. Neurogenin3, neurogenic differentiation, and MAF bZIP transcription factor A levels increased in NAS mice. CONCLUSION: Our study provides that Notch1 inhibition decreased insulin secretion and decreased islet and ß-cell masses. It is thought that Notch1 inhibition suppresses islet proliferation and induces differentiation of small islets. In conclusion, Notch signaling pathway may play an important role in ß-cell mass determination and diabetes.

Notch signaling pathway mediates the immunomodulatory mechanism of Yangfei Huoxue decoction alleviating bleomycin-induced pulmonary fibrosis in rats.

OBJECTIVE: To investigate the immunomodulatory mechanism by which Yangfei Huoxue decoction (YHD) alleviates bleomycin (BLM)-induced pulmonary fibrosis (PF) in rats. METHODS: Rats were randomly divided into two time-point groups (day 14 and 28), and each time-point group comprised the following six subgroups: control, BLM, dexamethasone (DXM), YHD high dose (YHD-H), YHD middle dose (YHD-M), and YHD low dose (YHD-L). Haematoxylin and eosin and Masson staining, flow cytometry, enzyme-linked immunosorbent assay, Western blotting and UPLC-QT of analyses were examined. RESULTS: The results showed that YHD reduced the degree of alveolar inflammation and fibrosis; downregulated the expression of CD28, CD80, CD86, Delta-like 1, Notch2, and Notch3; and upregulated the proportions of Th1/Th2 and Tc1/Tc2. The seven components of YHD were detected. CONCLUSION: The current study indicates that YHD mainly functions by regulating the immune system and that the molecular mechanism may be related to the regulation of the Notch signaling pathway.

High-content screen in human pluripotent cells identifies miRNA-regulated pathways controlling pluripotency and differentiation.

BACKGROUND: By post-transcriptionally regulating multiple target transcripts, microRNAs (miRNAs or miR) play important biological functions. H1 embryonic stem cells (hESCs) and NTera-2 embryonal carcinoma cells (ECCs) are two of the most widely used human pluripotent model cell lines, sharing several characteristics, including the expression of miRNAs associated to the pluripotent state or with differentiation. However, how each of these miRNAs functionally impacts the biological properties of these cells has not been systematically evaluated. METHODS: We investigated the effects of 31 miRNAs on NTera-2 and H1 hESCs, by transfecting miRNA mimics. Following 3-4 days of culture, cells were stained for the pluripotency marker OCT4 and the G2 cell-cycle marker Cyclin B1, and nuclei and cytoplasm were co-stained with Hoechst and Cell Mask Blue, respectively. By using automated quantitative fluorescence microscopy (i.e., high-content screening (HCS)), we obtained several morphological and marker intensity measurements, in both cell compartments, allowing the generation of a multiparametric miR-induced phenotypic profile describing changes related to proliferation, cell cycle, pluripotency, and differentiation. RESULTS: Despite the overall similarities between both cell types, some miRNAs elicited cell-specific effects, while some related miRNAs induced contrasting effects in the same cell. By identifying transcripts predicted to be commonly targeted by miRNAs inducing similar effects (profiles grouped by hierarchical clustering), we were able to uncover potentially modulated signaling pathways and biological processes, likely mediating the effects of the microRNAs on the distinct groups identified. Specifically, we show that miR-363 contributes to pluripotency maintenance, at least in part, by targeting NOTCH1 and PSEN1 and inhibiting Notch-induced differentiation, a mechanism that could be implicated in naïve and primed pluripotent states. CONCLUSIONS: We present the first multiparametric high-content microRNA functional screening in human pluripotent cells. Integration of this type of data with similar data obtained from siRNA screenings (using the same HCS assay) could provide a large-scale functional approach to identify and validate microRNA-mediated regulatory mechanisms controlling pluripotency and differentiation.

Notch-1 Inhibition Promotes Immune Regulation in Transplantation Via Regulatory T Cell-Dependent Mechanisms.

BACKGROUND: Transplantation is the treatment of choice for many patients with end-stage organ disease. Despite advances in immunosuppression, long-term outcomes remain suboptimal, hampered by drug toxicity and immune-mediated injury, the leading cause of late graft loss. The development of therapies that promote regulation while suppressing effector immunity is imperative to improve graft survival and minimize conventional immunosuppression. Notch signaling is a highly conserved pathway pivotal to T-cell differentiation and function, rendering it a target of interest in efforts to manipulate T cell-mediated immunity. METHODS: We investigated the pattern of Notch-1 expression in effector and regulatory T cells (Tregs) in both murine and human recipients of a solid-organ transplant. Using a selective human anti-Notch-1 antibody (aNotch-1), we examined the effect of Notch-1 receptor inhibition in full major histocompatibility complex-mismatch murine cardiac and lung transplant models, and in a humanized skin transplant model. On the basis of our findings, we further used a genetic approach to investigate the effect of selective Notch-1 inhibition in Tregs. RESULTS: We observed an increased proportion of Tregs expressing surface and intracellular (activated) Notch-1 in comparison with conventional T cells, both in mice with transplants and in the peripheral blood of patients with transplants. In the murine cardiac transplant model, peritransplant administration of aNotch-1 (days 0, 2, 4, 6, 8, and 10) significantly prolonged allograft survival in comparison with immunoglobulin G-treated controls. Similarly, aNotch-1 treatment improved both histological and functional outcomes in the murine lung transplant model. The use of aNotch-1 resulted in a reduced proportion of both splenic and intragraft conventional T cells, while increasing the proportion of Tregs. Furthermore, Tregs isolated from aNotch-1-treated mice showed enhanced suppressive function on a per-cell basis, confirmed with selective Notch-1 deletion in Tregs (Foxp3EGFPCreNotch1fl/fl). Notch-1 blockade inhibited the mammalian target of rapamycin pathway and increased the phosphorylation of STAT5 (signal transducer and activator of transcription 5) in murine Tregs. Notch-1low Tregs isolated from human peripheral blood exhibited more potent suppressive capacity than Notch-1high Tregs. Last, the combination of aNotch-1 with costimulation blockade induced long-term tolerance in a cardiac transplant model, and this tolerance was dependent on CTLA-4 (cytotoxic T-lymphocyte-associated antigen-4) signaling. CONCLUSIONS: Our data reveal a promising, clinically relevant approach for immune modulation in transplantation by selectively targeting Notch-1.

Inhibitory Effect of Gamma-Secretase on the Survival Rate of Dental Pulp Stem Cells: An in Vitro Study

Objective: To evaluate in vitro the effect of gamma-secretase inhibition on the survival of dental pulp stem cells. Material and Methods: Sound teeth have been used. Dental pulp stem cells were isolated by enzymatic digestion. An appropriate number of cells were treated with different concentrations of gamma secretase enzyme (DAPT) (1, 3, 6.25, 12.5, 25.5, 37.5, 50 and 100 µM). The metabolic activity of cells and the distribution of cells in different phages of cell cycle was evaluated by MTT assay and flow cytometry, respectively. Statistical analysis was made one-way ANOVA. Comparison was made between the groups on the level of p<0.05. Results: In low concentration of DAPT (1, 3, 6.25, 12.5) the growth rate of the cells increases, whereas in high concentration of DAPT (25.5, 37.5, 50, 100) can significantly reduce the viability of the treated cells. The results also indicate that DAPT can interrupt the cell cycle in G1 phase. Conclusion: The DAPT for dose-dependent survival rate of dental pulp stem cells and affect cell population increase in the G1 phase of the cell cycle.

Efficacy of Banxia Xiexin decoction in a rat model of chronic atrophic gastritis.

OBJECTIVE: To investigate the effectiveness of Banxia Xiexin decoction (BXD) in a rat model of chronic atrophic gastritis (CAG). METHODS: Sixty 6-week-old healthy Wistar rats (30 males, 30 females) were used in the present study. A rat model of CAG was successfully established using the combined active immunization/ethanol/sodium deoxycholate method. BXD was prepared from a mixture of seven Chinese herbs, and was intragastrically administered to CAG rats at three different doses (6, 12, and 24 g·kg－1·d－1). After 24 weeks, the rats were euthanized, and gastric tissue specimens were collected. Gastric mucosal specimens were stained with hematoxylin and eosin for histological examination to evaluate the degree of inflammation and morphological changes. Immunohistochemical staining was performed to examine the mucosal expression of proliferating cell nuclear antigen. Serum gastrin levels were measured using radioimmunoassay. The expression of Notch signaling-associated genes was examined by quantitative polymerase chain reaction and Western blot assay. RESULTS: BXD at all three doses significantly reversed the adverse effects generated by CAG in rats. Compared with control rats, the CAG rats who were administered BXD had an accelerated growth rate, reduced inflammatory cell infiltration, improved gastric mucosal morphology, augmented thickness of the gastric mucosa, increased number of gastric glands, enhanced mucosal expression of proliferating cell nuclear antigen, and elevated serum gastrin levels. CONCLUSION: BXD has a therapeutic effect in a rat model of CAG by targeting the Notch signaling pathway, thereby blocking the CAG from progressing to early gastric cancer.

Effect of Notch-Hes1 signaling blockade by a γ-secretase inhibitor on the expression of γδT17 cells in a mouse model of psoriasis-like skin inflammation / 中华皮肤科杂志

Objective@#To evaluate the effect of Notch-Hes1 signaling blockade by a γ-secretase inhibitor on the expression of γδT17 cells in a mouse model of psoriasis-like skin inflammation.@*Methods@#Forty-five healthy specific pathogen-free (SPF) male BALB/c mice were randomly divided into control group, model group and intervention group by simple random sampling. The model group and intervention group were both topically treated with imiquimod 5% cream (62.5 mg once a day) on the shaved back, the intervention group were then intraperitoneally injected with the γ-secretase inhibitor DAPT (10 mg/kg once a day) immediately after topical application of imiquimod, and the control group were topically treated with equivalent amount of vaseline once a day. After 6-day treatment, psoriasis area and severity index (PASI) was used to evaluate changes of skin lesions. On day 7, blood samples were obtained from all the mice through heart puncture after anesthetization, and spleen and skin tissues were acquired to prepare single cell suspension. Spleen index was compared among the 3 groups. Skin tissues on the mouse back were resected and subjected to hematoxylin-eosin staining to observe histopathological changes. Flow cytometry was performed to determine the percentage of γδT17 cells in the spleen and skin tissues, real-time reverse transcription (RT) -PCR to measure the mRNA expression of Hes1 in single cell suspension of the spleen, and enzyme-linked immunosorbent assay (ELISA) to determine the serum level of interleukin (IL) -17A. Statistical analysis was carried out by using one-way analysis of variance and repeated measures analysis of variance for comparison of indices among groups, and Pearson correlation analysis for evaluating the correlation between different indices.@*Results@#Twenty-four hours after the final treatment, the intervention group showed milder psoriasis-like skin inflammation, lower PASI score, and milder degree of epidermal thickening and dermal inflammatory cell infiltration compared with the model group. The model group showed significantly increased spleen index (12.534 ± 1.636) , proportions of γδT17 cells in the spleen (24.659% ± 4.603%) and skin tissues (22.127% ± 5.670%) , mRNA expression of Hes1 in the spleen (4.867 ± 0.543) , and serum level of IL-17A ([22.478 ± 2.776] ng/L) compared with the control group (all P < 0.01) . However, the above indices were significantly lower in the intervention group (9.449 ± 1.040, 14.966% ± 5.770%, 13.631% ± 5.946%, 2.541 ± 0.347, [18.639 ± 1.816] ng/L) than in the model group (all P < 0.01) . In the model group and intervention group, there were positive correlations between the proportions of γδT17 cells in the spleen and serum levels of IL-17A (r = 0.56, 0.53 respectively, both P < 0.05) , between the proportions of γδT17 cells in skin lesions and PASI scores (r = 0.56, 0.52 respectively, both P < 0.05) , as well as between the mRNA expression of Hes1 in the spleen and the proportions of γδT17 cells (r = 0.61, 0.58 respectively, both P < 0.05) or serum levels of IL-17A (r = 0.60, 0.54 respectively, both P < 0.05) .@*Conclusion@#Notch-Hes1 signaling blockade by γ-secretase inhibitor can markedly inhibit the expression of γδT17 cells, and effectively alleviate the severity of psoriasis-like skin inflammation in mouse models.

γ-Secretase mutations in acne inversa / 中华皮肤科杂志

Acne inversa is a chronic inflammatory skin disease of the folliculo-sebaceous-apocrine unit. Currently, genetic and immunological factors are hot topics in the study of its pathogenesis. Genetic factors are mainly related to γ-secretase mutations, and abnormal expression of the γ-secretase-Notch axis leads to increased keratinization of hair follicles and inflammation in some patients with haploinsufficiency of the γ-secretase gene. Mutations in the γ-secretase gene are not necessary for acne inversa, and the risk of Alzheimer′s disease in familial acne inversa patients still remains unclear. Some progress has been made in researches on the association of genotype with phenotype in acne inversa patients, but further studies with large sample size are needed for verification.

γ-Secretase mutations in acne inversa / 中华皮肤科杂志

Acne inversa is a chronic inflammatory skin disease of the folliculo-sebaceous-apocrine unit.Currently,genetic and immunological factors are hot topics in the study of its pathogenesis.Genetic factors are mainly related to γ-secretase mutations,and abnormal expression of the γ-secretase-Notch axis leads to increased keratinization of hair follicles and inflammation in some patients with haploinsufficiency of the y-secretase gene.Mutations in the γ-secretase gene are not necessary for acne inversa,and the risk of Alzheimer's disease in familial acne inversa patients still remains unclear.Some progress has been made in researches on the association of genotype with phenotype in acne inversa patients,but further studies with large sample size are needed for verification.

Expression of DLL3 in small cell lung cancer and its application in targeted therapy / 国际肿瘤学杂志

Small cell lung cancer (SCLC) has a poor biological behavior,high probability of recurrence and metastasis,and limited treatment.The Notch signaling pathway is an evolutionarily conserved pathway that regulates the growth of many cell types through local cell-cell interactions.It controls the differentiation,proliferation and survival of cells.As a ligand for the Notch pathway,delta-like protein 3 (DLL3) is highly expressed on the membrane of SCLC cells.DLL3 plays an important role in cancer initiation and epithelial mesenchymal transition,invasion and metastasis of SCLC.Rovalpituzumab tesirine is a conjugate of directed against DLL3,which shows great potential for SCLC therapy.

[Screening of microRNAs targeting Notch signaling pathway implicated in inner ear development and the role of microRNA-384-5p].

Objective: To screen the key microRNAs targeting Notch signaling pathway in inner ear and investigate its potential regulating function. Methods: The interaction network and the Core-Notch network, involved with key genes in Notch signal pathway and differential-expressed microRNAs in inner ear, were constructed by bioinformatics methods. The important microRNAs in regulating Notch signaling pathway were screened via topological and GO analysis, followed by in vivo and in vitro investigation. Results: MiRNA-384-5p was identified as a key regulator specifically expressed in mouse brain and inner ear, which could down-regulate Notch1. The Notch1 expression was found significantly down-regulated in miRNA-384-5p-mimic-transfected HeLa cells. The dual-luciferase reporter gene assay further confirmed the effect of miRNA-384-5p on the down-regulation of Notch1 and Dll4 in Notch signaling pathway. Conclusions: The Core-Notch network is constructed to screen microRNAs implicated in inner ear development, and miRNA-384-5p is screened and verified to be target-regulating the Notch signaling pathway, which could be the potential target in the regeneration of impaired hair cells.

Notch signaling pathway in lung cancer and lung cancer stem cells and its inhibitors / 国际肿瘤学杂志

Notch signaling pathway is involved in the abnormal differentiation and self-renewal of lung cancer stem cells.The further studies for the roles of Notch signaling pathway in the regulation of lung cancer stem cells are expected to find new targets in the diagnosis and treatment of lung cancer.Inhibitors of the Notch signaling pathway may be effective in the treatment of lung cancer.Lung cancer stem cells are thought to be a major cause of recurrence of lung cancer,therefore,targeted therapy for lung cancer stem cells may be more effective than treatment for the entire tumor.

Screening of microRNAs targeting Notch signaling pathway implicated in inner ear development and the role of microRNA-384-5p / 中华耳鼻咽喉头颈外科杂志

Objective@#To screen the key microRNAs targeting Notch signaling pathway in inner ear and investigate its potential regulating function.@*Methods@#The interaction network and the Core-Notch network, involved with key genes in Notch signal pathway and differential-expressed microRNAs in inner ear, were constructed by bioinformatics methods. The important microRNAs in regulating Notch signaling pathway were screened via topological and GO analysis, followed by in vivo and in vitro investigation.@*Results@#MiRNA-384-5p was identified as a key regulator specifically expressed in mouse brain and inner ear, which could down-regulate Notch1. The Notch1 expression was found significantly down-regulated in miRNA-384-5p-mimic-transfected HeLa cells. The dual-luciferase reporter gene assay further confirmed the effect of miRNA-384-5p on the down-regulation of Notch1 and Dll4 in Notch signaling pathway.@*Conclusions@#The Core-Notch network is constructed to screen microRNAs implicated in inner ear development, and miRNA-384-5p is screened and verified to be target-regulating the Notch signaling pathway, which could be the potential target in the regeneration of impaired hair cells.

Relationship between the Notch signaling pathway and the development and progression of liver fibrosis / 临床肝胆病杂志

The Notch signaling pathway mainly includes Notch receptors and ligands, transcription factors, and DNA binding proteins. It decides the way of cell proliferation, differentiation, and apoptosis. Recent studies have shown that the Notch signaling pathway plays an important role in the development and progression of liver fibrosis, and blocking or activating the Notch signaling pathway can influence the progression of liver fibrosis. This article reviews the research advances in the composition of the Notch signaling pathway, the mechanism by which it is activated, and its association with liver fibrosis.

Expression of Notch pathway receptors and ligands in cutaneous malignant melanoma / 中华皮肤科杂志

Objective To determine the expression of Notch pathway receptors (Notch1 and Notch4) and ligands (Jagged1 and Dll4) in cutaneous malignant melanoma (CMM) tissues,and to preliminarily explore the role of the Notch signaling pathway in the pathogenesis of CMM.Methods Immunohistochemical study was performed to determine the expression pattern and intensity of Notch1,Notch4,Jagged1 and Dll4 in 40 paraffin-embedded CMM specimens and 15 paraffin-embedded pigmented nevus specimens.Statistical analysis was carried out by chi-square test and Spearman rank correlation analysis with the SPSS 21.0 software.Results Notchl was detected in 31 (77.5％) of 40 CMM specimens,as well as in 3 of 15 pigmented nevus specimens,and the positive rates significantly differed between the two groups (x2 =15.281,P ＜ 0.001).However,no significant difference in the expression intensity of Notch1 was observed between 18 in situ melanoma tissues and 22 invasive melanoma tissues (x2 =0.631,P =0.427).In addition,the positive rates of Notch4,Jagged1 and Dll4 were also significantly higher in the CMM group than those in the pigmented nevus group (all P ＜ 0.05),and the expression intensity of Notch4,Jagged1 and Dll4 significantly differed between in situ and invasive melanoma tissues (all P ＜ 0.05).In CMM tissues,the expression of Notch1 was positively correlated with that of Jagged1 (rs =0.350,P =0.027) and Dll4 (rs =0.562,P ＜ 0.001),while the expression of Jaggedl was negatively correlated with that of Dl14 (rs =-0.734,P ＜ 0.001).Conclusion Abnormality of the Notch signaling pathway may be involved in the pathogenesis of melanoma,but further researches are still needed to elucidate the detailed mechanism.

Advances in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy / 国际脑血管病杂志

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common hereditary cerebral small vessel disease.NOTCH3 missense mutation causes its coded cysteine occurring odd change and then affects the conformation and function of protein of NOTCH3.The abnormal NOTCH3 protein has vascular smooth muscle toxicity and finally deposits in the cerebral small blood vessels and causes the disease.Usually,CADASIL can be suspected by its typical clinical manifestations and neuroimaging findings.Its diagnosis needs genetic testing or skin biopsy to find the outer granular osmiophilic deposits of small vascular smooth muscle cells or immunohistochemical NOTCH3-ECD staining positive.For nearly two decades,the studies on genetics,pathogenesis,clinical manifestations,and diagnostic techniques of CADASIL have made great progress,however,many important questions have not been fully clarified and have new discoveries,such as the NOTCH3 gene mutation pattern and loci,and the relationship between gene phenotype and clinical phenotype,optimization of diagnosis process,depth study of pathogenic mechanism,exploration of new discoveries,new therapeutic targets and concepts.This article reviews the genetic characteristics,pathogenesis,and clinical diagnosis and treatment technology of CADASIL.

R607C mutation of NOTCH3 gene and clinical features in 4 CADASIL families in Henan, China / 国际脑血管病杂志

Objective To investigate the NOTCH3 gene mutation and clinical features in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) families.Methods The clinical features of 4 CADASIL probands in Henan,China were analyzed retrospectively,and the incidences of other members in their families were investigated.The NOTCH3 gene mutations in the 3rd,4th,llth,and 18th exons were detected and the results were analyzed in the patients and some family members.Results Gene sequencing showed that 6 patients in 4 families and 1 mutant carrier had NOTCH3 gene R607C mutation in exon llth,they all met the clinical features of CADASIL.Three patients accompanied with vascular risk factors.The clinical stroke patients had unilateral limb weakness.All 5 patients with complete head MRIdata had thalamic infarction.Conclusions In the 4 CADASIL families of R607C mutation,the clinical features of 6 patients with CADASIL were similar,but there were individual differences in different family members.Imaging examination has important role in the diagnosis of CADASIL.The vascular risk factors,such as hyperte.

Expression analysis of notsh signaling pathway molecules in shed cultured in keratinocyte growth medium

Aim: To detect the expression of molecules associated with Notch signaling pathway in stem cells from human exfoliated deciduous teeth (SHED) cultured in specific differentiation medium, namely, keratinocyte growth medium (KGM). Methods:RNA was extracted from SHED harvested on day 1, 3 and 7. RNA was reverse-transcribed to obtain the cDNA and then proceeded with PCR using specific primers for the Notch signaling pathway molecules (Notch1, Jagged-1, Jagged-2 and, Hes1) as well as stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel and stained with SYBR green. Results:Notch-1 was highly expressed in SHED cultured in KGM and showed increase in density as the days progressed, while Jagged-1 showed a decrease. Jagged-2 on the other hand, showed a slight increase on day 3 followed by a decrease on day 7. However, Hes-1 was not expressed in SHED cultured in KGM. Nanog showed expression only on day 3 and gradually increased in expression on day 7. Conclusions:Notch signaling pathway associated molecules; Notch-1, Jagged-1, Jagged-2, and stem cell marker Nanog are expressed in SHED cultured in KGM which may be involved in the differentiation into epithelial-like cells in human dental pulp tissues.

Neuroprotective effect of cerebral ischemic preconditioning is associated with regulation of expressions of nuclear factor-κB and Hes1 in rats / 国际脑血管病杂志

Objective To investigate the neuroprotective mechanism of cerebral ischemic preconditioning by detecting the expression changes of hippocampus nuclear factor-κB (NF-κB) and Hesl mRNA after cerebral ischemia-reperfusion in rats.Methods A total of 108 healthy male SD rats were randomly divided into a cerebral ischemia group,a cerebral ischemic preconditioning group,and a sham operation group,and then redivided into 22 h,48 h,72 h,7 d,and 14 d subgroups.Ischemic preconditioning was performed at day 3 before establishing the cerebral ischemia-reperfusion injury model by transient occlusion of right internal carotid artery for 10 min.At each time point after cerebral ischemia-reperfusion,the neurological deficit score and cerebral infarction volume measurement were performed,and the expressions of NF-κB and Hes1 mRNA in the hippocampus were detected by using real-time fluorescence quantitative polymerase chain reaction.Results The neurological function scores and the percentage of cerebral infarction volume in the cerebral ischemic preconditioning goup at each time point were significantly lower than those in the cerebral ischemia-reperfusion group (all P ＜0.05).The expression levels of NF-κB and Hesl mRNAs in each group had progressive reduction with time.Compared with the same time point,it showed that the expression levels of NF-κB and Hes1 mRNAs in the cerebral ischemic preconditioning group and the cerebral ischemia-reperfusion group were significantly higher than those in the sham operation group,the expression level of NF-κB mRNA in the cerebral ischemic preconditioning group was significantly lower than that in the cerebral ischemia-reperfusion group,and the expression level of Hesl mRNA was significantly higher than that in the cerebral ischemia-reperfusion group (all P ＜0.05).Conclusions The upregulation of Hesl and down-regulation of NF-κB may be involved in the neuroprotective mechanisms of cerebral ischemic preconditioning.