Specific immediate early gene expression induced by high doses of salicylate in the cochlear nucleus and inferior colliculus of the rat / Expressão específica de genes precoces imediatos induzida por doses elevadas de salicilato no núcleo coclear e colículo inferior de rato

Abstract Introduction: Salicylate at high doses induces tinnitus in humans and experimental animals. However, the mechanisms and loci of action of salicylate in inducing tinnitus are still not well known. The expression of Immediate Early Genes (IEG) is traditionally associated with long-term neuronal modifications but it is still not clear how and where IEGs are activated in animal models of tinnitus. Objectives: Here we investigated the expression of c-fos and Egr-1, two IEGs, in the Dorsal Cochlear Nucleus (DCN), the Inferior Colliculus (IC), and the Posterior Ventral Cochlear Nucleus (pVCN) of rats. Methods: Rats were treated with doses known to induce tinnitus in rats (300 mg/kg i.p. daily, for 3 days), and c-fos and Egr-1 protein expressions were analyzed using western blot and immunocytochemistry. Results: After administration of salicylate, c-fos protein expression increased significantly in the DCN, pVCN and IC when assayed by western blot. Immunohistochemistry staining showed a more intense labeling of c-fos in the DCN, pVCN and IC and a significant increase in c-fos positive nuclei in the pVCN and IC. We did not detect increased Egr-1 expression in any of these areas. Conclusion: Our data show that a high dose of salicylate activates neurons in the DCN, pVCN and IC. The expression of these genes by high doses of salicylate strongly suggests that plastic changes in these areas are involved in the genesis of tinnitus.

Resumo Introdução: Salicilato em doses elevadas induz zumbido nos seres humanos e em animais experimentais. No entanto, os mecanismos e loci de ação do salicilato na indução de zumbido ainda não são bem conhecidos. A expressão dos genes precoces imediatos (GPIs) está tradicionalmente associada a alterações neuronais em longo prazo, mas ainda não está claro como e onde os GPIs são ativados em modelos animais de zumbido. Objetivos: No presente estudo investigamos a expressão de c-fos e Egr-1, dois GPIs, no núcleo coclear dorsal (NCD), colículo inferior (CI) e núcleo coclear ventral posterior (NCVp) de ratos. Métodos: Os ratos foram tratados com doses que, conhecidamente, induzem zumbido em ratos (300 mg/kg IP/dia, por três dias) e as expressões das proteínas c-fos e Egr-1 foram analisadas por meio de Western blot e imunoistoquímica. Resultados: Após a administração de salicilato, a expressão da proteína c-fos aumentou significativamente no NCD, NCVp e CI, quando analisados por Western blot. A coloração imunoistoquímica mostrou uma marcação mais intensa de c-fos no NCD, NCVp e CI e um aumento significativo de núcleos positivos de c-fos no NCVp e CI. Não detectamos aumento da expressão de Egr-1 em qualquer dessas áreas. Conclusão: Nossos dados mostram que uma dose alta de salicilato ativa neurônios no NCD, NCVp e CI. A expressão desses genes por doses altas de salicilato sugere que as alterações plásticas nessas áreas estão envolvidas na gênese do zumbido.

Association Study of FOS-Like Antigen-2 Promoter Polymorphisms With Papillary Thyroid Cancer in Korean Population

OBJECTIVES: FOS-like antigen-2 (FOSL-2), a member of the FOS gene family, encode leucine zipper proteins that can heterodimerize with proteins of Jun family. Thus, activating protein (AP)-1 transcription factor is formed, has a crucial role in proliferation, differentiation and apoptosis of normal tissue as well as oncogenic transformation and progression. We performed an association study of single nucleotide polymorphisms (SNPs) in the FOSL-2 with papillary thyroid cancer (PTC). We also estimated the relationships between the SNPs and the clinicopathologic characteristics of PTC. METHODS: One promoter SNPs (rs925255) of FOSL-2 gene were genotyped with direct sequencing method in 94 PTC and 213 controls. PTC patients were dichotomized and compared with respect to clinical parameters of PTC. Genetic data were analyzed using Helixtree, SNPAnalyzer, SNPStats. Multivariate logistic regression analysis was fulfilled to evaluate the genetic effect with adjustment for age and sex. RESULTS: SNP (rs925255) in FOSL-2 showed a significant association (codominant 1 model [G/G vs. A/G]: odds ratio [OR], 0.531, 95% confidence interval [CI], 0.293 to 0.96, P=0.036; dominant model: OR, 0.50, 95% CI, 0.28 to 0.89, P=0.015) with PTC. The frequency of allele G in rs925255 was also significantly associated with PTC (OR, 0.59; 95% CI, 0.34 to 0.91; P=0.02). But we fail to prove significant association between this polymorphism (rs925255) and clinico-pathological parameters. CONCLUSION: Our findings suggest that the rs925255 SNP and its allele G show significant association with the PTC in Korean population.

The relationship between c-fos gene and filamentous actin cytoskeleton in MG-63 osteoblasts under cyclic tensile stress / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between c-fos gene and filamentous actin (F-actin) in MG-63 osteoblasts under cyclic tensile stress.</p><p><b>METHODS</b>MG-63 osteoblasts were subjected to cyclic tensile stress (0.5 Hz, 2 000 microstrain) for 3, 6, and 12 h. The changes of c-fos gene were investigated by fluorescent quantitation polymerase chain reaction. Then the best loading time group was screened as the experimental group compared with 0 h group. The changes of F-actin and c-fos were investigated with or without cytochalasin D treatment.</p><p><b>RESULTS</b>Cyclic tensile stress induced high expression of c-fos mRNA, and peaked at 3 h. After loading, F-actin had a structure reorganization, but had no change in expression. After cytochalasin D treatment, the formation of stress fibers and the fluorescence intensity of F-actin cytoskeleton significantly reduced, meanwhile the c-fos mRNA expression was inhibited.</p><p><b>CONCLUSION</b>After loading, there is only structure reorganization for F-actin, and the expression has not any change. That means the remodeling F-actin is the existing one. F-actin reorganization is an important part in c-fos gene expression induced by stress.</p>

Effects of pyrroloquinoline quinone on proliferation and expression of c-fos, c-jun, CREB and PCNA in cultured Schwann cells / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of pyrroloquinoline quinine (PQQ) on proliferation and expression of c-fos, c-jun, CREB and PCNA in cultured Schwann cells.</p><p><b>METHODS</b>Schwann cells were cultured and purified in vitro. The purity of Schwann cells was identified by immunofluorescence of S-100. After synchronization of cell cycle by serum-free medium, different concentration of PQQ (0,1, 10, 100, 1,000, 10,000 nmol/L) were added into culture medium for 72 h. Flow cytometry was used to determine cell cycle. The content of c-fos, c-jun, and CREB mRNA were detected by RT-PCR, and the expression of PCNA protein was detected by Western blot.</p><p><b>RESULTS</b>After PQQ treatment, the percentage of cells in G0/G1 phase decreased and the percentage of cells in S and G2/M phase increased. After treated by PQQ at concentration of 1-10,000nmol/L, content of c-fos,c-jun,CREB mRNA was increased by 0.33,0.42 and 0. 52 fold (P < 0. 05). However, at concentration of 1 000 nmol/L, there was no difference in mRNAs content when compare to control (P >0.05). And it showed a decline at concentration of 10,000 nmol/L (P < 0.05). PCNA protein expression was up-regulated at PQQ concentration of 1-100 nmol/L. At 100 nmol/L, the expression increased by 1.17 fold (P < 0.05); However, at 1,000 nmol/L, there was no difference in PCNA expression when compared to control. And 10,000 nmol/L of PQQ inhibited the expression of PCNA (P < 0.05).</p><p><b>CONCLUSIONS</b>When treated with PQQ at concentration of 10-100 nmol/L, the proliferation of Schwann cells increased and the expression of c-fos,c-jun, CREB and PCNA was up-regulated.</p>

Gene expression pattern during osteogenic differentiation of human periodontal ligament cells in vitro

PURPOSE: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. METHODS: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 microg/mL ascorbic acid, 10 mM beta-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. RESULTS: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. CONCLUSIONS: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

Regulation of c-fos gene expression in hepatic stellate cells by transforming growth factor beta / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of transforming growth factor beta (TGF ) on c-fos gene expression in hepatic stellate cells.</p><p><b>METHODS</b>Hepatic stellate cells (HSC-T6) were cultured in the medium containing different concentrations of TGF (0.2, 1, and 5 ng/ml), and cells were collected at different time points of incubation (8, 24, 48, and 72 h). The total RNA of the HSCs was isolated and c-fos gene expression level were measured by reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>c-fos gene expression levels of HSCs cultured in the presence of low (0.2 ng/ml), moderate (1 ng/ml) and high (5 ng/ml) concentrations of TGF for 8, 24, 48 and 72 h were significantly greater than those of control group. The c-fos gene expression levels of HSCs increased gradually with the increment of TGF concentration, and significant differences in c-fos gene expression were found between the 3TGF groups.</p><p><b>CONCLUSION</b>TGF strongly up-regulates c-fos gene expression in hepatic stellate cells.</p>

Changes of c-fos, c-jun mRNA expressions in cardiomyocyte hypertrophy induced by angiotensin II and effects of tanshinone II A / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the changes of proto-oncogene c-fos, c-jun mRNA expression in angiotensin II (Ang II)-induced hypertrophy and effects of tanshinone II A (Tan) in the primary culture of neonatal rat cardiomyocytes.</p><p><b>METHOD</b>Twelve neonatal Wistar rats aged one day old of clean grade and both sexes were selected to isolate and culture cardiomyocytes. The cardiomyocytes were divided into: normal control group, Ang II (10(-6) mol x L(-1)) group, Ang II (10(-6) mol x L(-1)) +Tan (10(-8) g x L(-1)) group, Ang II (10(-6) mol x L(-1)) + valsartan (10(-6) mol x L(-1)) group, Tan (10(-8) g x L(-1)) group, valsartan (10(-6) mol x L(-1)) group. The cardiomyocyte size was determined by phase contrast microscope, the rate of protein synthesis in cardiomyocytes was measured by 3H-leucine incorporation. The c-fos, c-jun mRNA expression of cardiomyocytes were assessed using reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULT</b>Ang II was added to the culture medium and 30 min later, the c-fos, c-jun mRNA expression of cardiomyocytes increased significantly (P < 0. 01). After Ang II took effect for 24 h, the rate of protein synthesis in Ang II group increased more prominently than that in normal control group (P < 0.01). After Ang II took effect for 7 days, the size of cardiomyocyte in Ang II group increased obviously (P < 0. 05). If tanshinone II or valsartan was added to the culture medium before Ang II, both of them could inhibit the increase of c-fos, c-jun mRNA expression (P < 0.01), cardiomyocyte protein synthesis rate (P < 0.01), and cardiomyocyte size (P < 0.05) induced by Ang II.</p><p><b>CONCLUSION</b>Tanshinone II could ameliorate Ang II-induced cardiomyocytes hypertrophy by inhabiting c-fos, c-jun mRNA expression.</p>

The effects of superoxide dismutase on the expression of c-fos gene in the chronic post-ischemic pain model rats / 대한마취과학회지

BACKGROUND: Chronic post-ischemia pain (CPIP) model is reported to represent the complex regional pain syndrome type I. The administration of non-specific free radical scavengers reduced mechanical allodynia, but it is not evident which type of free radical is responsible for the development of CPIP. This study was investigated to elucidate the role of superoxide on the development of CPIP and the relationship with the expression of c-fos gene. METHODS: Male Sprague-Dawley rats weighing 290-310 g were housed in one cage with food and water ad libitum. CPIP model was made by placing a tourniquet on the left hindpaw of rats. The tourniquet maintained for 3 hours, then released to allow reperfusion. Thirty minutes before reperfusion, superoxide dismutase (SOD) or normal saline (control group) was injected. Mechanical allodynia and cold allodynia were measured at 1, 3, 5, 7, 14 and 28 days after reperfusion and compared. Also, spinal cord was harvested and the expression of c-fos gene was measured through the real time reverse transcription polymerase chain reaction. RESULTS: Superoxide dismutase reduced mechanical allodynia (1, 3, 5 and 14 day) and cold allodynia (1, 3 and 7 day) compared with control rats in left hindpaw. Expression of c-fos was significantly reduced in the SOD rats at the day 14 and 28 compare to the control rats. CONCLUSIONS: The administration of superoxide dismutase suppressed the allodynia and c-fos gene expression of CPIP model rats and it may be suggested that the superoxide has an important role in the development of CPIP.

Gene expression profiling of light-induced retinal degeneration in phototransduction gene knockout mice

Exposure to light can induce photoreceptor cell death and exacerbate retinal degeneration. In this study, mice with genetic knockout of several genes, including rhodopsin kinase (Rhok-/-), arrestin (Sag-/-), transducin (Gnat1-/-), c-Fos (c-Fos-/-) and arrestin/transducin (Sag-/-/Gnat1-/-), were examined. We measured the expression levels of thousands of genes in order to investigate their roles in phototransduction signaling in light-induced retinal degeneration using DNA microarray technology and then further explored the gene network using pathway analysis tools. Several cascades of gene components were induced or inhibited as a result of corresponding gene knockout under specific light conditions. Transducin deletion blocked the apoptotic signaling induced by exposure to low light conditions, and it did not require c-Fos/AP-1. Deletion of c-Fos blocked the apoptotic signaling induced by exposure to high intensity light. In the present study, we identified many gene transcripts that are essential for the initiation of light-induced rod degeneration and proposed several important networks that are involved in pro- and anti-apoptotic signaling. We also demonstrated the different cascades of gene components that participate in apoptotic signaling under specific light conditions.

Pim-1 kinase inhibits the activation of reporter gene expression in Elk-1 and c-Fos reporting systems but not the endogenous gene expression: an artifact of the reporter gene assay by transient co-transfection

We have studied the molecular mechanism and signal transduction of pim-1, an oncogene encoding a serine-threonine kinase. This is a true oncogene which prolongs survival and inhibits apoptosis of hematopoietic cells. In order to determine whether the effects of Pim-1 occur by regulation of the mitogen-activated protein kinase pathway, we used a transcriptional reporter assay by transient co-transfection as a screening method. In this study, we found that Pim-1 inhibited the Elk-1 and NFkappaB transcriptional activities induced by activation of the mitogen-activated protein kinase cascade in reporter gene assays. However, Western blots showed that the induction of Elk-1-regulated expression of endogenous c-Fos was not affected by Pim-1. The phosphorylation and activation of neither Erk1/2 nor Elk-1 was influenced by Pim-1. Also, in the gel shift assay, the pattern of endogenous NFkappaB binding to its probe was not changed in any manner by Pim-1. These data indicate that Pim-1 does not regulate the activation of Erk1/2, Elk-1 or NFkappaB. These contrasting results suggest a pitfall of the transient co-transfection reporter assay in analyzing the regulation of transcription factors outside of the chromosome context. It ensures that results from reporter gene expression assay should be verified by study of endogenous gene expression.

DNA damage, apoptosis and C-myc, C-fos, and C-jun overexpression induced by selenium in rat hepatocytes / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the effects of selenium on DNA damage, apoptosis and c-myc, c-fos, and c-jun expression in rat hepatocytes.</p><p><b>METHODS</b>Sodium selenite at the doses of 5, 10, and 20 micromol/kg was given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was detected by single cell gel electrophoresis (or comet assay). Hepatocellular apoptosis was determined by TUNEL (TdT-mediated dUTP nick end labelling) and flow cytometry. C-myc, c-fos, and c-jun expression in rat hepatocytes were assayed by Northern dot hybridization. C-myc, c-fos, and c-jun protein were detected by immunohistochemical method.</p><p><b>RESULTS</b>At the doses of 5, 10, and 20 micromol/kg, DNA damage was induced by sodium selenite in rat hepatocytes and the rates of comet cells were 34.40%, 74.80%, and 91.40% respectively. Results also showed an obvious dose-response relationship between the rates of comet cells and the doses of sodium selenite (r=0.9501, P<0.01). Sodium selenite at the doses of 5, 10, and 20 micromol/kg caused c-myc, c-fos, and c-jun overexpression obviously. The positive brown-yellow signal for proteins of c-myc, c-fos, and c-jun was mainly located in the cytoplasm of hepatocytes with immunohistochemical method. TUNEL-positive cells were detected in selenium-treated rat livers. Apoptotic rates (%) of selenium-treated liver cells at the doses of 5, 10, and 20 micromol/kg were (3.72 +/- 1.76), (5.82 +/- 1.42), and (11.76 +/- 1.87) respectively, being much higher than those in the control. Besides an obvious dose-response relationship between apoptotic rates and the doses of sodium selenite (r=0.9897, P<0.01), these results displayed a close relationship between DNA damage rates and apoptotic rates, and the relative coefficient was 0.9021, P<0.01.</p><p><b>CONCLUSION</b>Selenium at 5-20 micromol/kg can induce DNA damage, apoptosis, and overexpression of c-myc, c-fos, and c-jun in rat hepatocytes.</p>

Role of the Peripheral Vestibular System on Neuroplasticity Induced by Hypergravity Stimulation

BACKGROUND AND OBJECTIVES: Altered environmental gravity, including both hypo- and hypergravity, may result in space adaptation syndrome. To explore the characteristics of this adaptive plasticity, the expression of immediate early gene c-fos mRNA in the vestibular system following an exposure to hypergravity stimulus was determined in rats. MATERIALS AND METHOD: The animals were subjected to 2 G force (two-fold earth's gravity) stimulus for 3 hours, and were examined at post-stimulus hours 0, 2, 6, 12, and 24. Real time reverse transcription-polymerase chain reaction (RT-PCR) was adopted to analyze temporal changes in the expression of c-fos mRNA. RESULTS: The hypergravity stimulation produced the expression of c-fos mRNA in the vestibular ganglion, medial vestibular nucleus, inferior vestibular nucleus, hippocampus, vestibulocerebellum, and vestibular cortex. The peak expression occurred at hour 6 in the animals hypergravity-stimulated for 3 hours. Bilateral labyrinthectomy significantly attenuated the degree of up-regulation in c-fos mRNA expression. MK-801, an NMDA receptor antagonist, also significantly attenuated the degree of up-regulation in c-fos mRNA expression. CONCLUSION: These results indicate that the adaptive neuroplasticity in response to an altered gravity occurs in the vestibular-related organs in the central nervous system, in which peripheral vestibular receptors and NMDA receptors play an important role.

The Effect and Mechanism of Intravesical Electrical Stimulation in Spinal Cord Injured Rats / 대한비뇨기과학회지

PURPOSE: The objective of this study was to evaluate whether the effect on afferent c-fiber activity is the underlying mechanism of intravesical electrical stimulation (IVES) in spinal cord injured rats. MATERIALS AND METHODS: Thirty five female Sprague-Dawley rats weighting 200-300g each were divided into the normal and spinalized groups. For the spinalized rats, we observed the c-fos expression, and we compared this in the non-IVES group with that in the IVES group. Cystometrograms were performed for all the groups via a suprapubic catheter. RESULTS: After performing IVES in the normal and spinalized rats, the abnormal increases of the intercontraction interval (ICI) and the voiding pressure (VP) were reduced close to the normal range. In the spinalized rats, the number of c-fos positive cells in the dorsal commissure (DCM) decreased in the group that had IVES performed when compared with the non-IVES group. CONCLUSIONS: The IVES reduced the c-fos gene expression in the L6-S1 spinal cord segment and also the bladder hyperreflexia in the spinalized rats through the inhibition of afferent c-fiber activity, in addition to affecting the A delta mechanoreceptors.

Fos-like immunoreactivity in central nervous system of mice simultaneously exposed to the elevated plus-maze and nociception

It has been demonstrated that mice exhibit antinociception when they are exposed to the elevated plus-maze (EPM), an animal model of anxiety. To investigate which brain structures are activated during EPM exposure, the present study assessed the immunohistochemical staining for Fos-like immunoreactivity (Fos-LI) in mice intraperitoneally injected with saline or 0,6 por cente acetic acid (which produces nociception) and cofined in the open arm (threatening situation) or enclosed arm (Control) of the EPM. The following strutuctures were investigated: magnus, dorsal and median raphe nuclei (MR, DR and MnR), periaqueductal gray matter (PAG), dorsal and ventral hippocampus (DH and VH), amygdala (AMY), hypothalamus (HYP) and superior and inferior colliculi (SC and IC)...

Expressão neuroanatômica de c-Fos em ratos alimentados e em jejum após administração periférica aguda de fluoxetina / Neuroanatomical expression of c-Fos in fed and fasted rats after acute periferical fluoxetine administration

A fluoxetina é um inibidor seletivo de recaptação de serotonina que rapidamente promove aumento da disponibilidade sináptica deste neurotransmissor. A ativação do sistema serotoninérgico central pela fluoxetina, por sua vez, pode ser diferencialmente afetada pelo estado metabólico do animal. No presente estudo investigamos os efeitos da administração aguda de fluoxetina na expressão de c-Fos no cérebro de ratos em duas condições metabólicas distintas: alimentados e em jejum. DESENHO DO ESTUDO: Estudo tipo experimental. Ratos machos Wistar, pesando 220 ::t 30 g, foram sacrificados 2 horas após injeções intraperitoneais de salina (1 mVkg) ou fluoxetina (10 mg/kg). Após perfusão intracardíaca com solução tampão salina fosfato, seguida de paraformaldeído, os cérebros dos animais foram removidos, sendo posteriormente processada a imuno-histoquímica. A imunorreatividade para c-Fos foi quantificada por um sistema computadorizado (Image-Pro, Discovery). Tanto os animais tratados com fluoxetina no estado de jejum quanto no estado alimentado, apresentaram aumento significativo na expressão de c-Fos, comparados ao tratamento com salina, em algumas áreas hipotalâmicas, límbicas, circunventriculares, romboencefálicas e mesencefálicas. A comparação quantitativa dos dados obtidos dos animais alimentados e em jejum tratados com fluoxetina revelou que ratos no estado de jejum apresentaram expressão de c-Fos significativamente maior no hipotálamo ventromedial e núcleo paraventricular, comparados aos animais no estado alimentado...

Protective effects of ligustrazine against photoreceptor cell injury induced by N-methyl-N-nitrosourea and its mechanism / 药学学报

<p><b>AIM</b>To study the protective effect of ligustrazine against photoreceptor cell injury induced by N-methyl-N-nitrosourea (MNU) in Sprague-Dawley (SD) rats.</p><p><b>METHODS</b>Ligustrazine injections of different doses were injected intraperitoneally into 47-day female SD rats once a day and a single intraperitoneal injection of MNU 60 mg x kg(-1) was given to 50-day rats. At different intervals after MNU treatment,the animals were sacrificed. The apoptotic index of photoreceptor cells was calculated by TUNEL labeling at 24 h following MNU treatment; peripheral retinal damage was evaluated based on retinal thickness at the d 7 after MNU treatment, and the expression of c-jun and c-fos genes was detected by RT-PCR technique.</p><p><b>RESULTS</b>Ligustrazine injection could remarkably increase total thickness of peripheral retina and decrease apoptotic index of photoreceptor cells induced by MNU in a dose-dependent manner. Compared with MNU-treated rats, the gene expression of c-jun and c-fos was time-dependently down-regulated in ligustrazine-treated group.</p><p><b>CONCLUSION</b>Ligustrazine injection partially protects against MNU-induced retinal damage by down-modulating the expression of c-jun and c-fos genes to inhibit apoptosis of photoreceptor cells.</p>

Effect of electroacupuncture at "Neiguan" (PC 6) on expression of c-fos gene in myocardium of the rat with acute myocardial ischemia / 中国针灸

<p><b>OBJECTIVE</b>To explore the mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myocardial ischemia.</p><p><b>METHODS</b>The rats were randomly divided into a sham operation group, a myocardial ischemia model group and a myocardial ischemia model plus electroacupuncture group. The acute myocardial ischemia model was developed byligation of the descending anterior branch of the coronary artery, and electroacupuncture was given at bilateral "Neiguan" (PC 6). Serum myocardial enzymes was determined by biochemical method and the expression of c-fos mRNA in myocardium was detected by using reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The activities of serum myocardial enzymes and the expression of c-fos mRNA in ischemic myocardium were significantly increased as compared with those in the sham operation group (P < 0.05), and after electroacupuncture they were significantly decreased (P < 0.05).</p><p><b>CONCLUSION</b>The mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myochadial ischemia is possibly related with down-regulation of expression of c-fos mRNA in myocardium.</p>

Changes in the expression of apoptotic genes in the intestinal tissue of scalded rats before and after resuscitation / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the dynamic changes in the expression of c-fos, proliferation cell nuclear antigen (PCNA) and Bax in the intestinal tissue of scalded rats before and after resuscitation.</p><p><b>METHODS</b>Wistar rats inflicted with 30% TBSA full thickness scald were employed as the model and randomly divided into four groups with 8 in each group, i.e. 2.0, 2.5, 4.0, 6.0 postscald hour (PSH) groups. Rats in each group received routine fluid infusion at 2 PSH, and were sacrificed at 2, 2.5, 4, 6 PSH, respectively. Then the intestinal tissue of the rats was harvested for the detection of the expression of c-fos, PCNA and Bax.</p><p><b>RESULTS</b>The expression of c-fos, PCNA and Bax at 2.0 PSH group (65.8 +/- 4.2%, 74.5 +/- 2.4%, 26.3 +/- 5.7%, respectively) significantly increased when compared with those in 2.5 PSH group (92.4 +/- 5.7%, 85.6 +/- 4.5%, 67.1 +/- 6.6%, respectively) (P < 0.01). The expression of 3 genes increased dramatically at 2.5 and 4.0 PSH, and reached the peak at 2.5 PSH. There was no obvious difference in the gene expression between 4 PSH and 2 PSH groups.</p><p><b>CONCLUSION</b>The expression of apoptotic genes in the intestinal tissue of scalded rats increased significantly during early resuscitation stage after burn injury.</p>

The Subdivision of the Spinal Neurons for Detrusor Function / 대한비뇨기과학회지

Purpose: No ideal method for subdividing and assessing changes in neurons of the spinal cord during specific conditions has been established. We attempted to develop a method for subdividing spinal neurons using immunohistochemical and fluorescent staining, which is an important key towards understanding the mechanism of reflex voiding. Materials and Methods: Thirty Sprague-Dawley rats, weighting 200-300g, were divided into five groups. A cystometrogram was performed during saline or acetic acid instillation. We identified the neuronal pathway associated with the detrusor by injecting a pseudorabies virus (PRV) into the detrusor muscle and inspecting the changes in relation to different time sequences. An immunohistochemical staining method was used to stain the fos-protein encoded by the c-fos gene. Immunofluorescent staining was performed to evaluate changes in the neurons in relation to the voiding reflex, and the neurons then subdivided. Results: We confirmed pseudorabies virus (PRV) infection of the cells in the sacral parasympathetic nucleus through immunohistochemical staining two days after injection. On detection of an increase in c-fos positive cells after dividing the c-fos positive area of the L6 and S1 spinal cord into 4 sections, significant increases were observed in the sacral parasympathetic nucleus (SPN) and dorsal commissure (DCM). Double staining was performed to detect the neurons associated with the voiding reflex in the SPN and DCM areas showing overexpression of c-fos. Conclusions: The establishment of a method for detecting morphological changes, and subdividing neurons by immunohistochemical and fluorescent staining, may provide an important key towards understanding the mechanism of various neuromodulations of clinically applied treatments. (Korean J Urol 2005;46:487-494)

The effects of genistein on epidermal growth factor receptor mediated signal transduction pathway in human ovarian carcinoma cells lines SKOV3 and its xenograft in nude mice / 中华病理学杂志

<p><b>OBJECTIVE</b>It was reported previously that genistein could inhibit proliferation of human ovarian carcinoma cell line SKOV(3), but mechanism was not clear. There is a close relationship between EGFR mediated signal transduction pathway and the development of ovarian tumor. This study aimed to investigate the effects of genistein on the EGFR mediated signal transduction pathway and to clarify its mechanisms of proliferation inhibition on human ovarian carcinoma cell line SKOV(3) and its xenograft in nude mice.</p><p><b>METHODS</b>The expression of c-erbB-2 protein was determined using immunocytochemistry. The expressions of c-jun and c-fos protein were determined using Western blotting. The expression of c-erbB-2, c-raf-1, c-jun and c-fos mRNA were tested by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expression of c-erbB-2, c-raf-1 and its downstream gene c-jun and c-fos were decreased at mRNA level in the 20 micromol/L genistein group. The expression of c-erbB-2 protein were decreased, its average absorbency (A) were decreased after treatment of SKOV(3) with 20 micromol/L genistein for 48 h, reached at 0.42 +/- 0.02 (P < 0.05). Western blotting demonstrated that the expressions of c-jun and c-fos protein were decreased gradually after being treated with 20 micromol/L genistein for 12 - 72 h.</p><p><b>CONCLUSIONS</b>Genistein could down-regulate the expression of two key genes, c-erbB-2 and c-raf-1 at mRNA and protein level in the EGFR mediated signal transduction pathway, and down-regulate the expression of its downstream nuclear transcription factors c-jun and c-fos at mRNA and protein level. It is suggested that interfering the expressions of some key signal molecules in EGFR mediated signal transduction system by genistein may account for its molecular foundation of proliferation inhibition in ovarian carcinoma.</p>