Temporal pattern of Fos and Jun families expression after mitogenic stimulation with FGF-2 in rat neural stem cells and fibroblasts

Intense stimulation of most living cells triggers the activation of immediate early genes, such as Fos and Jun families. These genes are important in cellular and biochemical processes, such as mitosis and cell death. The present study focused on determining the temporal expression pattern of Fos and Jun families in fibroblasts and neural stem cells of cerebellum, hippocampus, and subventricular zone (SVZ) of rats of different ages at 0, 0.5, 1, 3, and 6 h after stimulation with fibroblast growth factor (FGF)-2. In neonates, a similar expression pattern was observed in all cells analyzed, with lower expression in basal condition, peak expression at 0.5 h after stimulation, returning to baseline values between 1 and 3 h after stimulation. On the other hand, cells from adult animals only showed Fra1 and JunD expression after stimulation. In fibroblasts and hippocampus, Fra1 reached peak expression at 0.5 h after stimulation, while in the SVZ, peak level was observed at 6 h after stimulation. JunD in fibroblasts presented two peak expressions, at 0.5 and 6 h after stimulation. Between these periods, the expression observed was at a basal level. Nevertheless, JunD expression in SVZ and hippocampus was low and without significant changes after stimulation. Differences in mRNA expression in neonate and adult animals characterize the significant differences in neurogenesis and cell response to stimulation at different stages of development. Characterizing these differences might be important for the development of cell cultures, replacement therapy, and the understanding of the physiological response profile of different cell types.

Major molecular events of reactivation of human cytomegalovirus after allogeneic hematopoietic stem cell transplantation / 器官移植

Immune deficiency of the host caused by allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the initial factor of reactivation of latent human cytomegalovirus (HCMV). The risk factors of reactivation of HCMV in allo-HSCT recipients consist of the serological status of HCMV in donors and recipients, the matching degree of human leukocyte antigen (HLA) and pretreatment patterns, etc. The reactivation of HCMV is associated with the expression of a series of viral cleavage and proliferation proteins induced by the overexpression of major immediate early promoter/enhancer (MIEP) in the viral genome. In this article, the risk factors of reactivation of HCMV after allo-HSCT, the molecular changes related to maintaining latent infection of HCMV, the key role of MIEP overexpression in reactivation of HCMV, and the molecular pathways involved in reactivation of HCMV after allo-HSCT were reviewed and the major molecular events of reactivation of HCMV after allo-HSCT were elucidated, aiming to provide reference for the prevention and treatment of cytomegaloviral disease (CMVD) after allo-HSCT.

Protective Effects of Ursolic Acid on Osteoblastic Differentiation via Activation of IER3/Nrf2 / 치위생과학회지

BACKGROUND: Oxidative stress is a known to be associated with in the pathogenesis of many inflammatory diseases, including periodontitis. Ursolic acid is a pentacyclic triterpenoid with has antimicrobial, antioxidative, and anticancer properties. However, the role of ursolic acid in the regulating of osteogenesis remains undetermined. This study was aimed to elucidate the crucial osteogenic effects of ursolic acid and its ability to inhibit oxidative stress by targeting the immediate early response 3 (IER3)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. METHODS: Cell proliferation was determined using water-soluble tetrazolium salt assay, cell differentiation was evaluated by alkaline phosphatase (ALP) activity, and formation of calcium nodules was detected using alizarin red S stain. Generation of reactive oxygen species (ROS) was determined using by DCFH-DA fluorescence dye in hydrogen peroxide (H₂O₂2)-treated MG-63 cells. Expression levels of IER3, Nrf2, and heme oxygenase-1 (HO-1) were analyzed using western blot analysis. RESULTS: Our results showed that ursolic acid up-regulated the proliferation of osteoblasts without any cytotoxic effects, and promoted ALP activity and mineralization. H₂O₂-induced ROS generation was found to be significantly inhibited on treatment with ursolic acid. Furthermore, in H₂O₂-treated cells, the expression of the early response genes: IER3, Nrf2, and Nrf2-related phase II enzyme (HO-1) was enhanced in the presence of ursolic acid. CONCLUSION: The key findings of the present study elucidate the protective effects of ursolic acid against oxidative stress conditions in osteoblasts via the IER3/Nrf2 pathway. Thus, ursolic acid may be developed as a preventative and therapeutic agent for mineral homeostasis and inflammatory diseases caused due to oxidative injury.

Illuminating the Activated Brain: Emerging Activity-Dependent Tools to Capture and Control Functional Neural Circuits / 神经科学通报·英文版

Immediate-early genes (IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to visualize and discriminate neural ensembles activated by multiple stimuli, and to map whole-brain-scale neural activation at single-neuron resolution. In addition, a collection of IEG-dependent molecular tools has been developed that can be used to complement the labeling of endogenous IEG genes and, especially, to manipulate activated neural ensembles in order to reveal the circuits and mechanisms underlying different behaviors. Here, we review these techniques and tools in terms of their utility in studying functional neural circuits. In addition, we provide an experimental strategy to measure the signal-to-noise ratio of IEG-dependent molecular tools, for evaluating their suitability for investigating relevant circuits and behaviors.

Baicalein Treatment Promotes Osteoblast Proliferation and Osteogenic Differentiation through Activation of Immediate Early Response 3 / 치위생과학회지

BACKGROUND: The primary aims of periodontal disease treatment is to remove dental plaque and calculus, the main causes of tooth loss, and restore periodontal tissue destroyed by inflammation. Periodontal disease treatment should also help maintain the alveolar bone, alleviate inflammation, and promote periodontal ligament cell proliferation, which is essential for tissue regeneration. Conventional antibiotics and anti-inflammatories have adverse side effects, especially during long-term use, so there is a need for adjunct treatment agents derived from natural products. The purpose of this study was to investigate whether the herbal flavone baicalein has the osteogenic activity under inflammatory conditions, and assess the involvement of osteoblast immediate early response 3 (IER3) expression.METHODS: Human osteoblastic MG-63 cells were cultured with the pro-inflammatory cytokines tumor necrosis factor α and interleukin 1β in the presence and absence of baicalein. Proliferation was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and expression of IER3 mRNA was assessed using real-time polymerase chain reaction. The expression of IER3 protein levels and activation of associated signal transduction pathways were assessed using western blotting.RESULTS: Baicalein increased IER3 mRNA and protein expression synergistically. In addition, baicalein reversed the suppression of cell proliferation, and the downregulation of osteogenic transcription factor runt-related transcription factor 2 and osterix induced by pro-inflammatory cytokines. Baicalein also upregulated the phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK 1/2). The upregulation of IER3 by pro-inflammatory cytokines was blocked by pretreatment with inhibitors of AKT, p38, JNK, and ERK 1/2.CONCLUSION: Baicalein mitigates the deleterious responses of osteoblasts to pro-inflammatory cytokines. Further, IER3 enhanced the effect of baicalein via activation of AKT, p38, JNK, and ERK pathways.

Preliminary screening of novel IER5 interacting proteins after ionizing irradiation / 中华放射医学与防护杂志

Objective To investigate the expression of radiation-induced IER5 protein and screen its potential interaction proteins that may participate in DNA repair process.Methods HeLa cells were irradiated with 4 Gy ionizing radiation.IER5 protein expression in whole cell lysate and in nuclear fraction were detected by Western blot at different timepoints after irradiation.3 × Flag-IER5 pCMV plasmid was constrcuted and the Flag tagged-IER5 expression was verified by Western blot.293T cells were transfected with 3 × Flag-IER5 pCMV plasmid.After irradiation the cells were collected and proteins were extracted.The IER5 interaction proteins were purified using immunoprecipitation and separated by 12％ SDS-polyacrylamide gel electrophoresis.Then the binding proteins were cut from the gel and analyzed by Mass spectrometry.Results The expression of IER5 protein began to increase 4 hour post-irradiation and its peak level was observed at 12 hour post-irradiation,and it lasted until 48 hour after irradiation.The expression level of IER5 protein in whole cell lysate and nuclear fraction were both increased.With the mass spectrometry analysis,a total of 347 proteins and 256 proteins were identified in irradiated and nonirradiated groups,respectively.Fourty one differential proteins were obtained,where 10 proteins were associated with DNA metabolic process and DNA rapair in the irradiated group and the poly(ADP-ribose) polymerase 1 (PARP1) protein was further confirmed by Western blot.Conclusions IER5 protein is an DNA damage related protein,and it may participate in DNA repair process.

In vitro antiviral mechanisms of eugeniin and quercetin against varicella-zoster virus / 中华微生物学和免疫学杂志

Objective To comparatively analyze the in vitro antiviral mechanism( s) of eugeniin and quercetin against varicella-zoster virus ( VZV) by using a novel antiviral assay based upon a reporter cell line (MV9G cells) for VZV. Methods Selection indexes (SIs) of potential antiviral compounds extracted from Chinese herbs or plants including eugeniin, eugenol, morin, curcumin, myricetin and quercetin for in vitro inhibition of VZV were calculated. The compounds with relatively higher SIs were screened out for fur-ther investigation of their in vitro inhibitory mechanisms with a cell-free virus ( CFVs) direct-infection assay and a cell-associated virus (CAVs) co-culture assay established with MV9G cells in our previous study. The inhibitory mechanisms analyzed in this study included direct inactivation of CFVs, inhibition of the adhesion and/or penetration capabilities of CFVs to MV9G cells, inhibition of the intracellular replication of CAVs and inhibition of the transcription and / or expression of viral immediate early gene 62 ( IE62 ) . Results Among the tested compounds, eugeniin and quercetin showed relatively higher SIs of 5. 82 and 8. 97, respec-tively. Eugeniin rather than quercetin directly but partly inactivated CFVs and inhibited their attachment to and penetration into MV9G cells in a concentration-dependent manner. Both eugeniin and quercetin revers-ibly inhibited the intracellular replication of CAVs and the transcription and expression of viral IE62 gene, for which eugeniin needed to be added within 12 hours after infection. Conclusion Eugeniin and quercetin had different in vitro inhibitory mechanisms against VZV, but inhibiting the transcription and expression of viral IE62 gene was a common mechanism shared by both of them.

Immediate expression of c-fos and c-jun mRNA in a model of intestinal autotransplantation and ischemia-reperfusion in situ

OBJECTIVE: Intestinal ischemia-reperfusion injury occurs in several clinical conditions and after intestinal transplantation. The aim of the present study was to investigate the phenomena of apoptosis and cell proliferation in a previously described intestinal ischemia-reperfusion injury autograft model using immunohistochemical markers. The molecular mechanisms involved in ischemia-reperfusion injury repair were also investigated by measuring the expression of the early activation genes c-fos and c-jun, which induce apoptosis and cell proliferation. MATERIALS AND METHODS: Thirty adult male Wistar rats were subjected to surgery for a previously described ischemia-reperfusion model that preserved the small intestine, the cecum and the ascending colon. Following reperfusion, the cecum was harvested at different time points as a representative segment of the intestine. The rats were allocated to the following four subgroups according to the reperfusion time: subgroup 1: 5 min; subgroup 2: 15 min; subgroup 3: 30 min; and subgroup 4: 60 min. A control group of cecum samples was also collected. The expression of c-fos, c-jun and immunohistochemical markers of cell proliferation and apoptosis (Ki67 and TUNEL, respectively) was studied. RESULTS: The expression of both c-fos and c-jun in the cecum was increased beginning at 5 min after ischemia-reperfusion compared with the control. The expression of c-fos began to increase at 5 min, peaked at 30 min, and exhibited a declining tendency at 60 min after reperfusion. A progressive increase in c-jun expression was observed. Immunohistochemical analyses confirmed these observations. CONCLUSION: The early activation of the c-fos and c-jun genes occurred after intestinal ischemia-reperfusion injury, and these genes can act together to trigger cell proliferation and apoptosis. .

Expressions of HIF-1αand CTGF proteins in oral submucous fibrosis / 中国医师杂志

Objective To evaluate the effects of hypoxia inducible factor-1 alpha ( HIF-1α) and connective tissue growth factor (CTGF) proteins in the pathogenesis of oral submucous fibrosis (OSF). Methods Expressions of HIF-1αand CTGF proteins in 30 cases of OSF, and 5 cases of normale control o-ral mucosa tissue were studied with streptomyces peroxidase ( SP) immunohistochemistry with the HIF-1αand CTGF rabbit anti-human polyclonal antibodies.Results The negative expressions of HIF-1αand CT-GF proteins were found in normal control and significant increase was found in OSF ( P <0.05).Conclu-sions HIF-1αand CTGF proteins have positive expression in OSF, both of them may act as an important role in pathogenesis of OSF.

Characterization of immediate early gene ORF62 of live attenuated varicella vaccine Oka strains pro-duced in China / 中华微生物学和免疫学杂志

Objective To investigate mutations in immediate early ( IE) gene ORF62 of three var-icella vaccine Oka strains ( vOka ) including two strains produced in China and their parental Oka strain (pOka), and then to further elucidate its possible roles in attenuation mechanism by comparing their ORF 62 promoter sequences and its activities , ORF62 coding regions and its transactivities .Methods ORF62 pro-moter-reporter plasmids and ORF62-expressing plasmids of pOka and three vOka strains ( vOka-BK from Changchun BCHT Biotechnology Co ., vOka-SH from Shanghai Institute of Biological Product Co .Ltd., and vOka-GSK from GlaxoSmithKline plc, as control) were constructed, respectively.ORF62 promoter regions and coding regions of the four strains were sequenced and then compared with each other .Differences of ac-tivities of the ORF62 promoter, and transactivities of the ORF62-encoded IE62 upon immediate early (ORF4), early (ORF28) and late (ORF67) gene promoters between pOka and vOka strains were assayed with transient transfection technique .Results Compared with pOka strain , three vOka strains had a con-sistent T deletion mutation at site 110 050 in ORF62 promoters, which did not result in any change of tran-scription factor binding motif .However , activities of ORF62 promoters from three vOka strains were signifi-cantly lower than those of pOka strain .Three consistent substitution mutations were observed in ORF 62 cod-ing regions of three vOka strains and three new enzyme restriction sites including SmaⅠ, NaeⅠand BssHⅡwere generated, respectively.Transactivities of IE62 from three vOka strains upon ORF4, ORF28 and ORF67 promoters were significantly higher than those of pOka both in CV-1 and MeWo cells , except that vOka-SH IE62 showed significantly lower transactivities upon ORF 4 promoter than those of pOka strain in CV-1 cells.Conclusion Consistent T deletion mutation at site 110 050 in ORF62 promoters of three vOka strains might be responsible for the reduced promoter activities and the changes of IE 62 transactivities .How-ever , it seemed that cell types have no significant effect on ORF 62 promoter activity or IE 62 transactivity be-tween pOka and vOka strains .

Investigation on inhibitory mechanisms of resveratrol on varicella-zoster virus in vitro / 中华微生物学和免疫学杂志

Objective To further investigate inhibitory mechanism(s) of resveratrol on varicellazoster virus (VZV) in vitro with our previously generated reporter cell line MV9G.Methods Cell-free VZVs were directly inoculated onto MV9G cells (CFVs direct-infection) or cell-associated VZVs wereco-cultured with MV9G cells (CAVs co-culture) to activate expression of reporter gene firefly luciferase in MV9G cells.Resveratrol was added before or after virus infection,roles of resveratrolon direct inactivation,on viral attachment to and penetration into MV9G cells,on intracellular viral replication and its IC50,inhibitorytime points and reversibility were assayed by comparing the luciferase activities reduction by resveratrol.Thereductions of VZV IE62 mRNA copies and IE62-antibody positive cells by resveratrol were further assayed.Results ATPs contents of MV9G cells in the presence of resveratrol over 30.0 μg/ml were concentrationdependently reduced,the CD50 of which was around 60.3 μg/ml.CFVs were premixed with 25.0 μg/ml resveratrol andincubated at 37℃ waterbath for two hours and then directly inoculated onto MV9G cells,luciferases activated by resveratrol-treated CFVs were reduced to around half of the untreated controls.MV9G cells were pre-incubated with resveratrol at 37℃ for 2 h and then directly infected with CFVs at 37℃ for another 2 h,the CFVs-activated luciferase was concentration-dependently reduced,but no big change was observed in those pre-incubated at 4℃.MV9G cells were co-cultured with CAVs in the presence of resvertrolfor 72 h,the CAVs-activated luciferases were markedly reduced in a concentration-dependent manner,the IC50 of which was around 8.7 μgml.Resveratrol was added in CAVs co-culture at 1,3,6,9,12,24,30,and 36 h post infection,the CAVs-activated luciferase in those resveratrol was added at 3,6,9,12,and 24 h post infection were significantly higher than those of controls.Resveratrol was withdrawn from CAVs coculture media,the CAVs-activated luciferases after withdrawal were significantly higher than those before,especially in those withdrswn at 24 and 72 h post infection.The IE62 mRNA levels shown by cDNA copiesdetected with SYBR Green RT-PCR and IE62 positive cells shown by monoclonal anti-IE62 antibody of thevirus-infected cells treated with resveratrol were significantly reduced with increase of incubation time withresveratrol.Conclusion Resveratrol was cytotoxic to MV9G cells,and the maximum resistant concentrationon MV9G cells was around 30.0 μg/ml,the CD50 of which was around 60.3 μg/ml.Non-cytotoxic resveratrol partly inactivated CFVs,inhibited viral penetration into rather than attachment to MV9G cells.Resveratrol inhibited CAVs' intmcellular replication strongly but reversibly in a concentration-dependent manner,the IC50 of which was around 8.7 μ/ml.The inhibition of resveratrol on VZV in vitro might be through suppression of IE62 gene transcription and expression in the early stage of infection.

Study on the binding sites of radiosensitivity associated transcription factor in the promoter region of Ier5 gene / 中华放射医学与防护杂志

Objective To clarify the mechanism of immediate early response gene 5 (ler5)transcription induced by radiation. Methods Deletant construction, site-specific mutagenesis,electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were used to forecast the promoter region,binding sites and transcription factors of Ier5 gene in HeLa cells.Results The promoter region of Ier5 gene might be in the region of Ier5 -8 deletant ( -408 - -238 bp).The Ier5 gene had two transcription factors of GCF and NFI,and GCF had two binding sites located in the region of - 388 - - 382 bp and - 274 - - 270 bp of Ier5 promoter.The binding site of NFI was located in - 362 --357 bp of Ier5 promoter. GCF could inhibit the expression of Ier5 gene and this inhibition was diminished when the radiation dose increased. In contrast, NFI increased the expression of Ier5.Conclusions The most possible region of Ier5 promoter is from -408 to - 238 bp which has two binding sites for the radiosensitivity transcription factors of GCF and NFI that could negatively and positively regulate the expression of Ier5 respectively.

Genetic association of the EGR2 gene with bipolar disorder in Korea

The early growth response gene 2 (EGR2) is located at chromosome 10q21, one of the susceptibility loci in bipolar disorder (BD). EGR2 is involved in cognitive function, myelination, and signal transduction related to neuregulin-ErbB receptor, Bcl-2 family proteins, and brain-derived neurotrophic factor. This study investigated the genetic association of the EGR2 gene with BD and schizophrenia (SPR) in Korea. In 946 subjects (350 healthy controls, 352 patients with BD, and 244 with SPR), nine single nucleotide polymorphisms (SNPs) in the EGR2 gene region were genotyped. Five SNPs showed nominally significant allelic associations with BD (rs2295814, rs61865882, rs10995315, rs2297488, and rs2297489), and the positive associations of all except rs2297488 remained significant after multiple testing correction. Linkage disequilibrium structure analysis revealed two haplotype blocks. Among the common identified haplotypes (frequency > 5%), 'T-G-A-C-T (block 1)' and 'A-A-G-C (block 2)' haplotypes were over-represented, while 'C-G-G-T-T (block 1)' haplotype was under-represented in BD. In contrast, no significant associations were found with SPR. Although an extended analysis with a larger sample size or independent replication is required, these findings suggest a genetic association of EGR2 with BD. Combined with a plausible biological function of EGR2, the EGR2 gene is a possible susceptibility gene in BD.

Preliminary study on Epstein-Barr virus infection in patients with drug eruption / 中华皮肤科杂志

Objective To explore the role of Epstein-Barr virus (EBV) infection in the etiology of drug eruption. Methods PCR-Southern blot was used to detect EBV-specific DNA fragment BamH I -W in peripheral blood mononuclear cells of 32 patients with drug eruption and 30 age- and sex-matched normal controls. The mRNA expression of EBV lyric gene BZLF1 in EBV DNA-positive samples was measured by RT-PCR and Southern blot. ELISA was performed to detect EBV virus capsule antigen (VCA)-specific IgM. Results The positivity rate of EBV DNA was significantly higher in patients with drug eruption than in normal controls (78.13% (25/32) vs 10.00% (3/30), P < 0.01), while no significant difference was noted between patients with severe and mild drug eruption (P > 0.05). The expression of BZLF1 mRNA was detected in 3 out of 25 EBV DNA-positive patients; of the 3 patients, 1 suffered from mild drug eruption, and 2 from severe drug eruption. EBV VCA-specific IgM was observed in 6 of 32 patients with drug eruption, but not in any normal controls. No significant difference in the positivity rate of EBV VCA-specific lgM existed between patients with severe and mild drug eruption (P > 0.05). Conclusions There is an active infection of EBV in patients with drug eruption. EBV infection is probably an environmental factor affecting the development of drug eruption.

Biological Analysis of HSV-1 Immediate-early Proteins ICPO, ICP22,and ICP27 in Neuro-blastoma Cells / 中国病毒学·英文版

The three immediate-early proteins of HSV-1, ICPO, ICP22, and ICP27, have specific and pivotal functions in transcriptional activation and inhibition, multiple regulatory and control processes of viral genes. In this paper, the expression and localization of these three proteins were studied in neuroblastoma cells using biochemical assays, and their possible and potential interactive functions are discussed. The data show that the three proteins are localized in different structures, specifically in the PML-NB-associated structure, which is a specific nuclear structure composed of many protein molecules and bound tightly to the nuclear matrix in neuroblastoma cells. The results suggest that the activating and suppressive functions of ICPs are mostly dependent on their transcriptional and regulatory roles, including the PML-NB-associated structure.

Expression of Activating Transcription Factor 3 in Ischemic Penumbra Region Following Focal Cerebral Ischemia / 대한해부학회지

It has been demonstrated that some of immediate early genes (IEGs) such as c-Jun or fos are induced immediately following neuronal injury and they play an important role in determining the fate of the injured neurons. Of IEGs, the activating transcription factor 3 (ATF3) is focused by many investigators, because they are expressed in various types of neural insults and have been known to serve a diverse function in both neuronal survival and death. However, little is known about the functional role of ATF3 in ischemic brain injury. So in this study, the authors examined the expression pattern of the activating transcription factor 3 (ATF3) following middle cerebral artery (MCA) occlusion-reperfusion injury. According to the findings obtained by triphenyltetrazolium chloride (TTC) stains, the authors have classified the infarcted area into two regions, the ischemic core region and the ischemic penumbra region. In both regions, many neurons underwent neuronal degeneration, characterized by the shrunken nuclei with eosinophilic perikaryon. The H & E stain also demonstrated the increased number of probable activated astrocytes and microglia in the ischemic brain regions and this was confirmed by GFAP- and OX42-immunohistochemistry. Immunohistochemical study for ATF3 also demonstrated the specific upregulation of ATF3 in the nuclei of neurons under ischemic injury, but not in those of the contralateral regions. Interestingly, the number of the ATF3 positive neurons in the ischemic penumbra regions outnumbered that of the ischemic core regions. Based on many reports that the neuronal death in ischemic penumbra region is caused by programed cell death rather than by necrosis which is main cause of neuronal death in ischemic core region, our results could suggest that the ATF3 is an important IEGs which determine the fate of the ischemic neurons.

Propofol-induced Immediate Early Gene Expression in Human Neuroblastoma Cell Lines / 대한마취과학회지

BACKGROUND: General anesthetics were known to induce expression of immediate early genes (IEGs), including c-fos and c-jun. However, mechanisms of IEG induction by general anesthetics were not fully understood. METHODS: IEG induction by propofol, a kind of intravenous anesthetics, and signal transduction pathways for propofol-induced IEG expression were investigated in human neuroblastoma cell line IMR32 and CHP134 with Northern and Western blot analysis. RESULTS: Cell viability was significantly decreased in IMR32 and CHP134 treated with increasing concentrations of propofol. IMR32 was more sensitive to propofol-induced cytotoxicity than CHP134. Propofol did not affect the cell cycle profile of IMR32. Expression of cyclin A, cyclin B1, CDK4 and CDK6 was increased in IMR32 by propofol treatment in a time-dependent manner. However, expression of cyclin A and CDK4 was decreased in CHP134. Proliferating cell nuclear antigen (PCNA) was increased in both IMR32 and CHP134 treated with propofol from 6 h to 24 h. c-fos and c-jun were induced by propofol treatment in both cells. Propofol also induced extracellular signal-regulated kinase (ERK) phosphorylation in both cells. Pretreatment of PD98059, an MEK inhibitor, blocked propofol-induced c-fos and c-jun expression.Propofol treatment was decreased nuclear transcription factor-kappa B (NF-kappa B) expression in IMR32, but not in CHP134. CONCLUSIONS: Propofol-induced c-fos expression might be mediated through ERK phosphorylation in both IMR32 and CHP134. Propofol-induced cytotoxicity, changes in expressions of cell cycle regulatory proteins, expression of IEGs, ERK phosphorylation, and NF-kappa B expression were different between IMR32 and CHP134.

A study on the mechanism of traumatic brain injury affecting the speed of bone fracture healing / 中华急诊医学杂志

Objective To evaluate the possible mechanism of traumatic brain injury (TB1) affecting the speed of bone fracture healing.Method TBI combined with unilateral tibial fracture (group A) was used to build multiple injury model and simple unilateral tibial fracture (group B),and the FOS,JUN,bFGF,and VEGF protein expression in different time points between the two groups were compared,and roentgenogram was used for the evaluation of bone healing.Results The expression of FOS,JUN,bFGF,and VEGF protein of the cerebral tissue was low in the normal rats,but was slightly enhanced in group B.There was consistence of development for FOS and JUN expression in the brain tissue in group A,reaching peak at post-TBI 3 hours,and then reducing to control level after 12 hours.The bFGF and VEGF reached peak at post-TBI 12 hours and 24 hours and reduced to control level after 72 hours,respectively.In group A and group B,an increase in the FOS,JUN protein expression around the fracture site was observed at 3 hours after injury,which reached the peak at 6 hours,and reduced to the control level after 24 hours;the comparison between group A,group B and the control group at 3 hours,6 hours and 12 hours had significant difference (P

Detection of Cytomegalovirus by Dual-PCR / 대한임상미생물학회지

BACKGROUND: Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality in transplant recipients and immunocompromised patients. We compared the results of a dual polymerase chain reaction (dual-PCR) and an antigenemia (Ag) test for detection of CMV from blood samples. METHODS: Between February 2002 and May 2005, we analyzed 175 blood samples submitted for CMV tests at Hanyang University Hospital. The late antigen (LA) and major immediate early (MIE) genes of CMV were concurrently amplified in the dual-PCR. The lower matrix protein pp65 of CMV was detected for the Ag test (Chemicon, Temecula, CA, USA). RESULTS: The positive rate of the dual-PCR was 14.3% (25/175) and that of the Ag test was 13.1% (23/175). The concordance rate of the dual-PCR and Ag test was 85.1% (149/175), while the discordance rate was 14.9% (26/175). CONCLUSION: The dual-PCR is a useful method for the early detection of CMV, but we recommend using both the dual-PCR and Ag test for detection of CMV due to a high discordance rate of the two methods.

Expression of immediate early gene following N-methyl-N-nitrosourea-induced retinal degeneration in SD rats / 中国药理学通报

Aim To study the molecular mechanism of retinal toxicity induced by N-methyl-N-nitrosourea (MNU) in SD rats. Methods A single intraperitoneal injection of 60 mg?kg~(-1) MNU was given to 50-day-old female SD rats. After MNU treatment for different times, the rats were sacrificed and both eyes were enucleated immediately and processed for histological examination. The apoptotic index of photoreceptor cells was calculated by TUNEL labeling and the expression of c-jun and c-fos genes was detected by RT-PCR technique. Results After the application of MNU for 24 h,the disorientation of photoreceptor outer segments was seen. The outer nuclear layer and photoreceptor layer were almost completely lost at 7 d. Apoptosis had already started at 12 h post-MNU and peaked at 24 h. MNU time-dependently up-regulated the expression of c-jun and c-fos genes. Conclusion MNU has a toxic effect on retina by up-modulating expression of c-jun and c-fos genes to promote photorecptor cells apoptosis.