The Impacts of Simulated Microgravity on Rat Brain Depended on Durations and Regions / 生物医学与环境科学（英文）

OBJECTIVE@#To explore the dynamic impacts of simulated microgravity (SM) on different vital brain regions of rats.@*METHODS@#Microgravity was simulated for 7 and 21 days, respectively, using the tail-suspension rat model. Histomorphology, oxidative stress, inflammatory cytokines and the expression of some key proteins were determined in hippocampus, cerebral cortex and striatum.@*RESULTS@#21-day SM decreased brain derived neurotrophic factor and induced neuron atrophy in the cerebral cortex. Strong oxidative stress was triggered at day 7 and the oxidative status returned to physiological level at day 21. Inflammatory cytokines were gradually suppressed and in striatum, the suppression was regulated partially through c-Jun/c-Fos.@*CONCLUSION@#The results revealed that the significant impacts of SM on rat brain tissue depended on durations and regions, which might help to understand the health risk and to prevent brain damage for astronauts in space travel.

JNK/c-Jun signaling pathway mediates arginine vasopressin neuron regeneration by promoting cytoskeleton reconstruction in rats with electrical lesions of the pituitary stalk / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which doublecortin promotes the recovery of cytoskeleton in arginine vasopressin (AVP) neurons in rats with electrical lesions of the pituitary stalk (PEL).@*METHODS@#Thirty-two SD rats were randomized into PEL group with electrical lesions of the pituitary stalk through the floor of the skull base (=25) and sham operation group (=7), and the daily water consumption (DWC), daily urine volume (DUV) and urine specific gravity (USG) of the rats were recorded. Four rats on day 1 and 7 rats on each of days 3, 7 and 14 after PEL as well as the sham-operated rats were sacrificed for detection of the expressions of β-Tubulin (Tuj1), doublecortin and caspase- 3 in the AVP neurons of the supraoptic nucleus using immunofluorescence assay and Western blotting.@*RESULTS@#After PEL, the rats exhibited a typical triphasic pattern of diabetes insipidus, with the postoperative days 1-2 as the phase one, days 3-5 as the phase two, and days 6-14 as the phase three. Immunofluorescent results indicated the repair of the AVP neurons evidenced by significantly increased doublecortin expressions in the AVP neurons following PEL; similarly, the expression of Tuj1 also increased progressively after PEL, reaching the peak level on day 7 after PEL. The apoptotic rates of the AVP neurons exhibited a reverse pattern of variation, peaking on postoperative day 3 followed by progressive reduction till day 14. Western blotting showed that the expressions of c-Jun and p-c-Jun were up-regulated significantly on day 3 ( < 0.05) and 7 ( < 0.01) after PEL, while an upregulated p-JNK expression was detected only on day 3 ( < 0.05), as was consistent with the time-courses of neuronal recovery and apoptosis after PEL.@*CONCLUSIONS@#JNK/c-Jun pathway is activated after PEL to induce apoptosis of AVP neurons in the acute phase and to promote the repair of neuronal cytoskeleton by up-regulation of doublecortin and Tuj1 expressions.

Alterations in gene expression profiles in children with Type 2 diabetes mellitus and its mechanisms / 中南大学学报(医学版)

To explore the mechanisms for Type 2 diabetes mellitus (T2DM) in children and provide genomic evidence for its early diagnosis and treatment.
 Methods: The peripheral blood gene chip datasets from 12 children with T2DM and 24 healthy children were retrieved from the Gene Expression Omnibus (GEO) at National Center for Biotechnology Information (NCBI). The differentially expressed genes were screened by R language software. GenCLiP 2.0, STRING, and Cytoscape software were used to analyze the biological functions, protein-protein interaction network, signal pathway, gene-pathway network, expression of key genes, and predictive value between the two differentially expressed genes.
 Results: A total of 79 differentially expressed genes were identified. Among them, 58 (73.42%) were up-regulated, and 21 (26.58%) were down-regulated. Differentially expressed genes mainly involved molecular functions and biological processes, such as defensive response, response to external stimulus, and inflammatory responses. At the same time, they were mainly involved in the Leishmaniasis, cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway. interleukin 1β (IL-1β), jun proto-oncogene (JUN), and IL-8 were 3 important linking nodes in the protein-protein interaction network. JUN and IL-1β were key genes, which were related to interleukin 17 (1L-17) signaling pathway, Toll-like receptor signaling pathway and so on. The expression of JUN gene in peripheral blood of children with T2DM was decreased while the expression of IL-1β gene was increased. JUN and IL-1β genes possessed certain diagnostic and predictive value in children with T2DM.
 Conclusion: The gene expression profile of peripheral blood in children with T2DM changes significantly. The genes of JUN and IL-1β are closely related to T2DM in children. IL-1β gene expression level shows a better predictive value on T2DM in children.

Effect of Inhibiting and Activating Wnt Signalling Pathway on NSC67657-inducing Monocytic Differentiation of HL-60 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of inhibiting and activating Wnt signalling pathway on monocyte differentiation of HL-60 cells induced with a new steroidal drug NSC67657 and its possible mechamism.</p><p><b>METHODS</b>The HL-60 cells were treated with 5, 10 and 20 µmol/L XAV-939 (inhibitor of Wnt signalling pathway) for 3 days, and with 10, 20 and 30 mmol/L LiCl (activator of Wnt signalling pathway) for 1 day; the expression levels of down-stream genes and proteins of Wnt signolling pathway were detected by RT-PCR and Western blot, respectively; the expression of cell surface differentiation antigen CD14 and early apoptosis of HL-60 cells was detected by flow cytometry, moreover the most suitable concentration of Wnt inhibitor and activator for HL-60 cells was determined. Then the HL-60 cells with inhibited and activated Wnt pathway were treated with NSC67657 of 10 µmol/L for 3 days; the expression levels of CD14 and down-stream target proteins of Wnt signalling pathway in blank control (culture mediam) group, simple NSC67657-treated group, NSC67657 combined with inhibitor group and NSC67657 combined activator group were compared and analyzed.</p><p><b>RESULTS</b>20 µmol/L XAV-939 and 20 mmol/L LiCl could effectively inhibit and activate Wnt signalling pathway of HL-60 cells respectively, could significantly down- and up-regulate the expression of cyclinD1, TCF1 and c-Jun genes (P < 0.05) and proteins (P < 0.05); moreover, the number of CD10(+) HL-60 cells in these conditions was below 1%, no early apoptosis of HL-60 cells was found. In the simple NSC67657-treated groups, the expression of cyclinD1, TCF1 and c-Jun proteins was down-regulated (P < 0.05), and the percentage of CD14(+) HL-60 cells accounted for 62.13 ± 9.44; after the HL-60 cells were treated with XAV-939, the NSC67657 could more significantly down-regulate the expression of cyclinD1, TCF1 and c-Jun proteins and the percentage of CD14(+) HL-60 cell accounted for 84.17 ± 5.39%, as compared with simple NSC67657-treated group; as compared with blank controls group, the expression of cyclinD1, TCF1 and c-Jun proteins was more obviously down-regulated and the percentage of CD14(+) HL-60 cells decreased to 33.99 ± 8.37% in NSC67657 combined LiC1 streated group, but which were higher than those in simple NSC67657-treated group (P < 0.05).</p><p><b>CONCLUSION</b>20 µmol/L XAV-939 and 20 mmol/L LiCl as effective inhabitor and activator of Wnt signalling pathway respectively can significantly down- and up-regulate the expression of Wnt down-stream pathway target genes and proteins. The influence of XAV-939 and LiC1 on differentiation of HL-60 cells induced by NSC67657 suggests that Wnt signalling pathway plays a key role in monocyte differentiction of HL-60 cells induced by NSC67657.</p>

Ultraviolet A Enhances Cathepsin L Expression and Activity via JNK Pathway in Human Dermal Fibroblasts / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cathepsin L (CatL) is a cysteine protease with strong matrix degradation activity that contributes to photoaging. Mannose phosphate-independent sorting pathways mediate ultraviolet A (UVA)-induced alternate trafficking of CatL. Little is known about signaling pathways involved in the regulation of UVA-induced CatL expression and activity. This study aims to investigate whether a single UVA irradiation affects CatL expression and activity and whether mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway is involved in the regulation of UVA-induced CatL expression and activity in human dermal fibroblasts (HDFs).</p><p><b>METHODS</b>Primary HDFs were exposed to UVA. Cell proliferation was determined by a cell counting kit. UVA-induced CatL production and activity were studied with quantitative real-time reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and fluorimetric assay in cell lysates collected on three consecutive days after irradiation. Time courses of UVA-activated JNK and p38MAPK signaling were examined by Western blotting. Effects of MAPK inhibitors and knockdown of Jun and Fos on UVA-induced CatL expression and activity were investigated by RT-PCR, Western blotting, and fluorimetric assay. Data were analyzed by one-way analysis of variance.</p><p><b>RESULTS</b>UVA significantly increased CatL gene expression, protein abundance, and enzymatic activity for three consecutive days after irradiation (F = 83.11, 56.14, and 71.19, respectively; all P < 0.05). Further investigation demonstrated phosphorylation of JNK and p38MAPK activated by UVA. Importantly, inactivation of JNK pathway significantly decreased UVA-induced CatL expression and activity, which were not affected by p38MAPK inhibition. Moreover, knockdown of Jun and Fos significantly attenuated basal and UVA-induced CatL expression and activity.</p><p><b>CONCLUSIONS</b>UVA enhances CatL production and activity in HDFs, probably by activating JNK and downstreaming AP-1. These findings provide a new possible molecular approach for antiphotoaging therapy.</p>

Expressiona of c-Jun and collagens I and III in cultured human skin fibroblasts are affected by infrared ray radiation / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of solar infrared ray (IR) radiation on the expressions of c-Jun and collagens I and III in cultured human skin fibroblasts (HSFs) and explore the molecular mechanism by which IR radiation causes aging of the skin.</p><p><b>METHODS</b>Primarily cultured HSFs exposed to IR radiation were examined for changes of the cell viability with MTT assay. The mRNA and protein expressions of c-Jun and collagens I and III was detected with real-time quantitative PCR and immunocytochemistry.</p><p><b>RESULTS</b>MTT assay showed that IR irradiation caused inhibition of cell proliferation compared with the control cells. The mRNA and protein expression of collagen I was decreased significantly by IR irradiation with the increase of the irradiation dose (P<0.01). HSFs irradiated by IR for 12 h showed a dose-dependent reduction of the expression of collagen type III mRNA and protein (P<0.05, P<0.01), but the expression increased dose-dependently in response to IR exposure for 24 h (P<0.05 or 0.01). IR irradiation enhanced the mRNA and protein expression of c-Jun in a dose-dependence manner (P<0.05 or 0.01).</p><p><b>CONCLUSIONS</b>IR irradiation can increase the expression of c-Jun, inhibit the expression of collagen I, and cause disturbance in collagen III expression in human skin fibroblasts, which may be one of the mechanism of IR radiation to initiate and promote skin photoaging.</p>

Hericium erinaceus (Bull.: Fr.) Pers., a medicinal mushroom, activates peripheral nerve regeneration / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the ability of aqueous extract of Hericium erinaceus mushroom in the treatment of nerve injury following peroneal nerve crush in Sprague-Dawley rats.</p><p><b>METHODS</b>Aqueous extract of Hericium erinaceus was given by daily oral administration following peroneal nerve crush injury in Sprague-Dawley rats. The expression of protein kinase B (Akt) and mitogen-activated protein kinase (MAPK) signaling pathways; and c-Jun and c-Fos genes were studied in dorsal root ganglia (DRG) whereas the activity of protein synthesis was assessed in peroneal nerves by immunohistochemical method.</p><p><b>RESULTS</b>Peripheral nerve injury leads to changes at the axonal site of injury and remotely located DRG containing cell bodies of sensory afferent neurons. Immunofluorescence studies showed that DRG neurons ipsilateral to the crush injury in rats of treated groups expressed higher immunoreactivities for Akt, MAPK, c-Jun and c-Fos as compared with negative control group (P <0.05). The intensity of nuclear ribonucleoprotein in the distal segments of crushed nerves of treated groups was significantly higher than in the negative control group (P <0.05).</p><p><b>CONCLUSION</b>H. erinaceus is capable of promoting peripheral nerve regeneration after injury. Potential signaling pathways include Akt, MAPK, c-Jun, and c-Fos, and protein synthesis have been shown to be involved in its action.</p>

Activation of NF-κB and AP-1 Mediates Hyperproliferation by Inducing β-Catenin and c-Myc in Helicobacter pylori-Infected Gastric Epithelial Cells

PURPOSE: In the gastric mucosa of Helicobacter pylori (H. pylori)-infected patients with gastritis or adenocarcinoma, proliferation of gastric epithelial cells is increased. Hyperproliferation is related to induction of oncogenes, such as β-catenin and c-myc. Even though transcription factors NF-κB and AP-1 are activated in H. pylori-infected cells, whether NF-κB or AP-1 regulates the expression of β-catenein or c-myc in H. pylori-infected cells has not been clarified. The present study was undertaken to investigate whether H. pylori-induced activation of NF-κB and AP-1 mediates the expression of oncogenes and hyperproliferation of gastric epithelial cells. MATERIALS AND METHODS: Gastric epithelial AGS cells were transiently transfected with mutant genes for IκBα (MAD3) and c-Jun (TAM67) or treated with a specific NF-κB inhibitor caffeic acid phenethyl ester (CAPE) or a selective AP-1 inhibitor SR-11302 to suppress activation of NF-κB or AP-1, respecively. As reference cells, the control vector pcDNA was transfected to the cells. Wild-type cells or transfected cells were cultured with or without H. pylori. RESULTS: H. pylori induced activation of NF-κB and AP-1, cell proliferation, and expression of oncogenes (β-catenein, c-myc) in AGS cells, which was inhibited by transfection of MAD3 and TAM67. Wild-type cells and the cells transfected with pcDNA showed similar activities of NF-κB and AP-1, proliferation, and oncogene expression regardless of treatment with H. pylori. Both CAPE and SR-11302 inhibited cell proliferation and expression of oncogenes in H. pylori-infected cells. CONCLUSION: H. pylori-induced activation of NF-κB and AP-1 regulates transcription of oncogenes and mediates hyperproliferation in gastric epithelial cells.

Effects of β-catenin on differentially expressed genes in multiple myeloma / 华中科技大学学报（医学）（英德文版）

This study aimed to identify the differentially expressed genes after silencing of β-catenin in multiple myeloma transduced with β-catenin shRNA. The DNA microarray dataset GSE17385 was downloaded from Gene Expression Omnibus, including 3 samples of MM1.S (human multiple myeloma cell lines) cells transduced with control shRNA and 3 samples of MM1.S cells transduced with β-catenin shRNA. Then the differentially expressed genes (DEGs) were screened by using Limma. Their underlying functions were analyzed by employing Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. Moreover, DEGs annotation was conducted based on the databases of tumor associated genes, tumor suppressed genes and the transcriptional regulation from patterns to profiles. Furthermore, the protein-protein interaction (PPI) relationship was obtained from STRING and the protein-protein interaction network and the functional modules were visualized by Cytoscape. Then, the pathway enrichment for the DEGs in the functional module was performed. A total of 301 DEGs, including 124 up-regulated and 117 down-regulated DEGs, were screened. Functional enrichment showed that CCNB1 and CDK1 were significantly related to the function of cell proliferation. FOS and JUN were related to innate immune response-activating signal transduction. Pathway enrichment analysis indicated that CCNB1 and CDK1 were most significantly enriched in the pathway of cell cycle. Besides, FOS and JUN were significantly enriched in the Toll-like receptor signaling pathway. FOXM1 was identified as a transcription factor. Moreover, there existed interactions among CCNB1, FOXM1 and CDK1 in PPI network. The expression of FOS, JUN, CCNB1, FOXM1 and CDK1 may be affected by β-catenin in multiple myeloma.

Antinociceptive Effect of Najanalgesin from Naja Naja Atra in a Neuropathic Pain Model via Inhibition of c-Jun NH2-terminal Kinase / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Najanalgesin, a toxin isolated from the venom of Naja naja atra, has been shown to exert significant analgesic effects in a neuropathic pain model in rats. However, the molecular mechanism underlying this protective effect of najanalgesin is poorly understood. The present study sought to evaluate the intracellular signaling pathways that are involved in the antinociceptive effect of najanalgesin on neuropathic pain.</p><p><b>METHODS</b>The antinociceptive properties of najanalgesin were tested in hind paw withdrawal thresholds in response to mechanical stimulation. We analyzed the participation of the mitogen-activated protein kinase p38, extracellular-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) by western blot analysis. This inhibition of JNK was confirmed by immunohistochemistry.</p><p><b>RESULTS</b>The phosphorylation levels of JNK (as well as its downstream molecule c-Jun), p38, and ERK were significantly increased after injury. Najanalgesin only inhibited JNK and c-Jun phosphorylation but had no effect on either ERK or p38. This inhibition of JNK was confirmed by immunohistochemistry, which suggested that the antinociceptive effect of najanalgesin on spinal nerve ligation-induced neuropathic pain in rats is associated with JNK activation in the spinal cord.</p><p><b>CONCLUSION</b>The antinociceptive effect of najanalgesin functions by inhibiting the JNK in a neuropathic pain model.</p>

La transexualidad: aspectos jurídico-sanitarios en el ordenamiento español / Transsexuality: law and health-related aspects in the Spanish legal system

El fenómeno de la transexualidad es un asunto en el que el peso social, en concreto de los colectivos transexuales, ha sido y sigue siendo crucial en muchos aspectos, desde la progresiva eliminación de la discriminación hasta la influencia para que el poder legislativo se pronuncie. En este artículo de investigación se tratará especialmente una de las reivindicaciones clásicas del colectivo, esto es, el tratamiento sanitario integral de la persona transexual dentro del Sistema Nacional de Salud. En este sentido, se observarán los avances en el desarrollo de un sistema sanitario adecuado para este colectivo, su tratamiento por parte de los distintos ordenamientos jurídicos en España, en general, y en alguna de sus comunidades autónomas con legislaciones más destacables (en especial Andalucía como comunidad autónoma pionera, el País Vasco y la Comunidad Foral de Navarra) y los retos pendientes, haciendo una especial investigación en torno a las sustanciales novedades que ha implantado en este ámbito la publicación de la quinta edición del Manual diagnóstico y estadístico de los trastornos mentales.

The social weight of transsexual groups has been and continues to be crucial in many aspects regarding transsexuality, from the progressive elimination of discrimination to influence in the legislative branch. This paper especially discusses a classic demand of these groups, comprehensive medical treatment of transsexual people within the National Health System. Thus, progress in the development of an adequate healthcare system for these groups, their treatment in the legal systems of Spain in general and of some of its autonomous communities with more noteworthy laws (especially in Andalusia, an autonomous community that has been pioneering in this regard, as well as the Basque Country and Navarre) and remaining challenges will be observed in this work. The article will also take particular note of the substantial developments that the publication of the Fifth Edition of the Diagnostic and Statistical Manual of Mental Disorders has established in this area.

The rescue effect of RANKL on zoledronate induced acid inhibition of osteoclastogenesis and gene expression of NF-kappaB p50 and c-Jun / 生物医学工程学杂志

In this study, the rescue effect of receptor activator for nuclear factor-kappaB ligand (RANKL) on zoledronate acid (ZOL) induced inhibition of osteoclastogenesis and gene expression of NF-kappaB p50 and c-Jun was investigated. Mice calvarial osteoblasts (OBs) were harvested and co-cultured with RAW264.7 cells and the cells were divided into 4 groups and received treatment with ZOL and RANKL, either single or combined. The formation of multi-nucleated osteoclast (OC) was examined and gene expression of NF-kappaB p50 and c-Jun was detected. Group B (ZOL) showed least multi-nucleated OC and resorption lacunae among the 4 groups (P < 0.05 or P < 0.01) and it was followed by group C (ZOL+RANKL). Group D (RANKL) showed highest OC and resorption lacunae while it was similar to Group A (control) (P > 0.05). Gene expression of NF-kappaB p50 and c-Jun was the lowest in group B (P < 0.05 or P < 0.01) among the four groups and was significantly increased in group C when compared with group B (P < 0.05). Group A and D showed highest gene expression and they were similar to each other (P > 0.05). This study suggest that RANKL might partly rescue ZOL induced inhibition of osteoclastogenesis, and the effect of RANKL and ZOL on osteoclastogenesis may be mediated by NF-kappaB p50 and c-Jun.

Hypoxia-induced autophagy contributes to radioresistance via c-Jun-mediated Beclin1 expression in lung cancer cells / 华中科技大学学报（医学）（英德文版）

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.

The effect of forsythiaside on the expression of c-jun induced by cisplatin in the cochlea of guinea pig / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the effect of forsythiaside on the expression of c-jun induced by cisplatin in the cochlea of guinea pig.@*METHOD@#Thirty guinea pigs were randomly divided into control group (10), cisplatin group (10) and forsythiaside group (10). The ototoxicity model was done with intraperitoneal injection of cisplatin solution (8 mg/kg per day) for 7 days. Forsythiaside (25 mg/kg per day) was injected 30 min before cisplatin solution treated in guinea pigs of forsythiaside group for 7 consecutive days. The saline instead of cisplatin was injected in normal control group. The distortion product otoacoustic emission (DPOAE) was detected before animals were killed. The expression of c-jun in cochlea of guinea pigs was detected by western blotting. The expression of c-jun mRNA in cochlea of guinea pigs was detected by reverse transcriptase polymerase chain reaction (RT-PCR).@*RESULT@#DPOAE amplitudes in cisplatin group was significantly lower than in control group (P < 0.01). Compared with cisplatin group, DPOAE amplitudes in forsythiaside group was increased significantly (P < 0.05). The expression of c-jun protein and mRNA were significantly increased in cisplatin group than in control group (P < 0.01). Compared with cisplatin group, the expression of c-jun protein and mRNA were significantly decreased in forsythiaside group.@*CONCLUSION@#Forsythiaside can significantly reduce the side effects induced by cisplatin through down-regulating the expression of c-jun.

Sinalização de morte celular via NA+, K+-ATPase como alvo de drogas anticâncer: estudos com o álcool perílico / Signaling of cell death via NA+, K+ -atpase as a target of anticancer drugs: studies with perillyl alcohol

A Na+, K+ ATPase (NKA) atua na manutenção do potencial de membrana dascélulas e em mecanismos de transdução de sinal. Alterações na atividade da NKAsão importantes em muitos processos biológicos e patológicos. A NKA pode serinibida pelo álcool perílico (POH), um monoterpeno utilizado no tratamento detumores, incluindo cerebrais. Neste trabalho, determinamos que o POH tambématua sobre cascatas de sinalização, moduladas via NKA, que controlam aproliferação e/ou morte celular. Foi avaliado o efeito do POH e do PA seu principalmetabólito, sobre a atividade da NKA em duas linhagens de células de glioblastoma(GBM) humano (U87 e U251), células não tumorais de astrócitos de camundongo eda linhagem (VERO). Nossos resultados, baseados na avaliação da atividade daNKA por incorporação do Rb+ , o qual mimetiza o K+ , mostraram uma sensibilidade àinibição pelo POH semelhante entre os quatro tipos de células (IC50 U87 2 mM;U251 1,8 mM; VERO 2,4 mM e astrócitos de camundongo 1,4 mM), enquanto o PAnão apresentou efeito. Sabe-se que nos GBMs existe uma superexpressão dasubunidade α1 da NKA, situada na estrutura das cavéolas que é provavelmenteresponsável pelo papel sinalizador atribuído a essa enzima, especialmente emrelação aos mecanismos apoptóticos. Comparamos a viabilidade celular,determinando a atividade da enzima lactato desidrogenase presente nosobrenadante das células tratadas por 24 h com POH e PA. O PA não alcançouefeito citotóxico igual ou superior a 30 por cento nas células mesmo na concentraçãoelevada de 4 mM. Já o POH reduziu, de maneira dependente da concentração, aviabilidade das células (IC50 U87 1,1 mM; U251 1,4 mM; VERO 0,9 mM e astrócitosde camundongo 1,4 mM). Na análise por western blot, 1,5 mM de POH ativou aproteína c-Jun N-terminal quinase (JNK), nas células U87, U251 e nos astrócitos decamundongo (incubação de 30 min)...

The Na+, K+ ATPase (NKA) acts in keeping the cell membrane potential and in signaltransduction mechanisms. Modifications in the activity of this enzyme are important inphysiological and pathological processes. The NKA is inhibited by perillyl alcohol(POH), a monotherpene used in the treatment of tumors, including brain tumors. Inthis work, we also show that POH acts in signaling cascades associated to NKA,controlling cell proliferation and/or cellular death. We evaluated the effect of POH andof its main metabolite (perillic acid - PA) on the NKA activity in cultured glioblastomacells (GBM) U87 and U251 and on non-tumor cells (mouse astrocytes and VEROcells). NAK activity was measured by non-radioactive Rb+incorporation by cells (Rb+is a K+substitute). Our results showed a similar sensitivity for the four cells typestested (IC50 U87 – 2 mM; U251 - 1,8 mM; VERO - 2,4 mM and mouse astrocytes -1,4 mM). Perillic acid did not show any effect in any cell type. In GBMs, it is knownthat NKA α1 subunit is super expressed. This isoform is embedded in caveolarstructures and is probably responsible by the signaling properties of this enzyme inapoptosis mechanisms. Cell viability was measured by lactate dehydrogenase in cellsupernatants of POH treated cells. The maximum PA cytotoxic effect obtained was30 percent even at 4 mm. However, POH reduced dose dependently cell viability, (IC50 U87- 1,1 mM; U251 - 1,4 mM; VERO - 0,9 mM and mouse astrocyte - 1,4 mM).Considering the western blot analysis, 1,5 mM POH activated the c-Jun N-terminalKinase (JNK), on U87, U251 and in mouse astrocytes after 30min incubation...

Inhibitory effect of PD98059 on MAPK signaling pathway in acute lymphocytic leukemia cells / 中国实验血液学杂志

This study was purposed to investigate the effect of blocking Ras/Erk signaling pathway on expression of important transcription factor c-fos, c-jun and TAK1 gene in primary acute lymphocytic leukemia (ALL) cells. The best effective concentration and effect time of PD98059 were screened; the expression levels of c-fos, c-jun and TAK1 in primary cultured cells of normal persons, primary cultured ALL cells and primary cultured ALL cells treated by PD98059 were detected by SYBR GreenI real-time quantitative-PCR. The results showed that before treatment by PD98059 the expression levels of c-fos and TAK1 mRNA were significantly up-regulated in primary cultured ALL cells as compared with primary cultured cells of normal persons (P = 0.014 and P = 0.017 respectively). After treatment by PD98059, the expression levels of c-fos, c-jun mRNA decreased in all 7 serum samples, while expression of TAK1 was down-regulated in 5 samples, and up-regulated in 2 samples. After treatment with PD98059, there was no statistical difference of c-fos, c-jun and TAK1 expression levels in primary cultured ALL cells and primary cultured normal cells. It is concluded that the c-fos and TAK1 activity of primary cultured ALL cells increases, and blocking the Ras/Erk signaling pathway of ALL cells can lead to obvious decrease of important transcription factors c-fos, c-jun, TAK1 genes expression.

S632A3 promotes LPS-induced IFN-beta production through inhibiting the activation of GSK-3beta / 药学学报

LPS stimulation of macrophages production of IFN-beta plays a key role in innate immunity defending the microbial invasion. In this study, the effect of S632A3 promoting LPS-induced IFN-beta production and the underlying mechanism were investigated, mRNA level was measured by real-time PCR, cytokine production was determined by ELISA, GSK-3beta activity was investigated by kinase assay, protein phosphorylation and expression were evaluated by Western blotting. The results revealed that S632A3 significantly augmented IFN-beta production by LPS-stimulated macrophages. S632A3 inhibition of the activation of GSK-3beta, reduced the threonine 239 phosphorylation of transcription factor c-Jun but increased the total level of c-Jun in LPS-stimulated macrophages. Moreover, small interfering RNA-mediated knockdown of c-Jun level abrogated the ability of S632A3 to augment IFN-beta. The study thus demonstrates S632A3 being a new anti-inflammation lead compound and provides a molecular mechanism by which S632A3 promoted LPS-induced IFN-beta production in macrophages through inhibiting the activation of GSK-3beta.

Expressions of p-c-jun and cyclinD1 between vascular smooth muscle cells and endothelial cells exposured to cigarette smoke extract / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the cell viabilities of vascular smooth muscle cells and vascular endothelial cells stimulated by cigarette smoke extract(CSE) .</p><p><b>METHODS</b>The CSE was prepared by smoke-bubbled phosphate buffered saline(PBS) generation.After culturing cells with different concentrations of CSE, we used the cell counting kit-8 to determine the cell viability.The expression levels of c-jun and cyclinD1 were analyzed through Western blot.The c-jun plasmid was transfected to detect the change of cyclinD1 expression.</p><p><b>RESULTS</b>The smooth muscle cell viability increased when the CSE concentration ranged 0.625%-10%, whereas the endothelial cells viability decreased when exposed to the CSE concentration. After exposure to CSE for 48 hours, there was no difference in c-jun expression between toxin group and PBS group;however, the expression of p-c-jun in the smooth muscle cells significantly increased in the toxin groups than in the PBS group(P<0.05) and the expression of p-c-jun in the vascular endothelial cells significantly decreased(P<0.05) . The level of cyclinD1 significantly increased after exposed to CSE, and its expression level also increased in respond to the c-jun overexpression.</p><p><b>CONCLUSION</b>CSE can enhance the proliferation of vascular smooth muscle cells and decrease in the activity of endothelial cells proliferation, which may be explained by the phosphorylation of c-jun and the expression of cyclinD1.</p>

Effects of glutamine induced heat shock protein 70 overexpression on atrial fibrosis and connexin 43 remodeling in isoprenaline-treated rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the effects of glutamine (Gln) induced heat shock protein 70(Hsp70) overexpression on atrial fibrosis and connexin 43 remodeling in isoprenaline(ISO)treated rats and related mechanisms.</p><p><b>METHODS</b>Forty male SD rats were randomly divided into five groups (n = 8 each group): control group, DMSO group, ISO 5 mg×kg(-1)×d(-1) group (Fibrosis group), ISO 5 mg×kg(-1)×d(-1) + Ala-Gln 0.75 mg×kg(-1)×d(-1) group (Intervention group) and ISO 5 mg×kg(-1)×d(-1) + QUE 100 mg× kg(-1)×d(-1) + Ala-Gln 0.75 mg×kg(-1)×d(-1) + DMSO group (QUE group).Rats were killed after 7 d. The AngII expression in myocardial tissue was detected by radioimmunoassay; myocardial fibrosis was observed by HE staining.Collagen volume fractions were quantified by Masson staining and as the indicators of atrial fibrosis. The expressions of Hsp70, p-JNK1/2/3, c-Jun and Cx43 were determined with immunohistochemical method.</p><p><b>RESULTS</b>AngII content was similar between the control group [(68.51 ± 10.76) pg/L] and DMSO [(71.47 ± 11.49) pg/L] group (P > 0.05), and significantly increased in fibrosis group [(211.25 ± 49.49) pg/L], intervention group [(185.32 ± 54.85) pg/L] and QUE [(189.90 ± 42.12) pg/L] group (P < 0.01 vs. control group). Atrial fibrosis was significantly higher in the fibrosis group [(29.485 ± 9.966)%] and QUE group [(25.060 ± 8.581)%] but not in the intervention group [(7.861 ± 1.867)%] compared to control group [(6.842 ± 1.674)%] and DMSO group [(7.108 ± 1.343)%]. The expression of Hsp70 was similar among the control group (0.160 ± 0.023), DMSO group (0.163 ± 0.022), fibrosis group (0.166 ± 0.028) and QUE (0.168 ± 0.027) group (P > 0.05) while significantly upregulated in the intervention group (0.215 ± 0.018) (P < 0.01 vs. control group). The expressions of p-JNK1/2/3 and c-Jun were similar between control group (0.151 ± 0.016;0.163 ± 0.022) and DMSO group (0.154 ± 0.021;0.164 ± 0.024)(P > 0.05), while significantly upregulated in fibrosis group (0.202 ± 0.025; 0.254 ± 0.044) and QUE group (0.196 ± 0.024; 0.251 ± 0.027) (P < 0.01 vs. control group) but not in intervention group (0.160 ± 0.025; 0.168 ± 0.024)were not changed obviously (P > 0.05 vs. control group). The content of Cx43 was similar between control group and DMSO group (0.231 ± 0.035 vs. 0.220 ± 0.032, P > 0.05), and was linearly distributed in intercalated disc of the cardiomyocytes, however, the content of Cx43 was significantly reduced (P < 0.01) and the Cx43 distribution was disordered in fibrosis group (0.163 ± 0.013) and QUE group (0.165 ± 0.024), while these changes were not found in intervention group.</p><p><b>CONCLUSION</b>Glutamine could reduce the atrial fibrosis and Cx43 remodeling in isoprenaline-treated rats by up-regulating Hsp70 and inhibiting JNK signaling pathway activation through down-regulating p-JNK1/2/3 and c-Jun expression.</p>

Expression and significance of MMP-7, c-Jun and c-Fos in rats skin photoaging

OBJECTIVE@#To investigate the expression and significance of the MMP-7, c-Jun and c-Fos in rat photoaging skin.@*METHODS@#A total of 45 SD rats were randomly divided into control group, model group, natural recovery group, physiological saline injection group and dermal pluripotent stem cells transplantation (DMSCs group), model group, natural recovery group, physiological saline injection group. DMSCs were treated with UV lamp irradiation to establish light aging skin model. Rats were then sacrificed after model prepared, no treatment was processed in the natural recovery group. Saline injections was adopted in saline group, DESCs group was treated with DESCs transplantation. Rats were sacrificed after 4 weeks. The expression of MMP-7, c-Jun and c-Fos were detected using the immunohistochemical method.@*RESULTS@#In model group, MMP 7 positive expression was higher than that in the other 4 groups, but without statistically difference (P>0.05); c-Jun, c-Fos expression were higher than that in the control group and DESCs group (P0.05).@*CONCLUSIONS@#MMP-7, c-Jun and c-Fos can be used as diagnosis indicators in the early stage of light aging, and they jointly participate in its development. DMSCs transplants is effective in treating light aging skin.