Identification of the effect of cisplatin on the transcriptome of human hepatocellular carcinoma cell lines / 基础医学与临床

Objective To investigate the effect of cisplatin treatment on the transcriptional level of human liver cancer cells by conducting transcriptome sequencing analysis after treating human liver cancer cell lines with differ-ent concentrations of cisplatin(CDDP).Methods Liver cancer cell lines HepG2 and Huh7 were incubated with cisplatin at different final concentrations of 0,20,50,100 and 200 μmol/L.After 12 hours,cell viability,immuno-fluorescence and RNA-sequencing(RNA-seq)were performed.Differential gene expression analysis(DEG),KEGG pathway analysis,and protein-protein interaction network analysis were conducted.Results Cisplatin de-creased cell viability and increased DNA damage in HepG2,Huh7 cells.Among the genes regulated after cisplatin treatment at different concentrations,59 genes were commonly up-regulated in both HepG2 and Huh7 cells,while 81 genes were commonly down-regulated.The commonly upregulated genes were mainly enriched in cancer initiation and progression pathways.The 81 commonly down-regulated genes were mainly enriched in Rap1 signaling pathway,Ras signaling pathway,signaling pathways regulating pluripotency of stem cells,axon guidance,and cell adhesion-related pathways.Survival analysis of key nodes in the protein-protein interaction network of commonly up-regulated and downregulated genes revealed a significant correlation between high expression of Jun proto-oncogene,AP-1 transcription factor subunit(JUN)and prolonged patient survival and a significant correlation between low ex-pression of growth arrest and DNA damage inducible alpha(GADD45A)and prolonged patient survival.Conclu-sions The study revealed common transcriptional changes in liver cancer cells under cisplatin treatment.Differential expression of JUN and GADD45A is a potential core mechanism to explain drug resistance.This conclusion provides some important prognostic indicators for clinical treatment.

Effect of cyclin D1 on HBV transcription and replication / 临床肝胆病杂志

Objective To investigate the effect of cyclin D1 (with CCND1 as the gene name) on HBV replication and its potential mechanism. Methods With reference to GSE84044 dataset, the Spearman's rank correlation analysis was used to investigate the correlation between the expression levels of genes in liver tissue and serum HBV DNA load in patients with HBV-related liver fibrosis. Cyclin D1 and cyclin D1 T286A mutant were transiently expressed in the HBV cell replication model, and time-resolved immunofluorescence and quantitative real-time PCR were used to measure the levels of HBsAg/HBeAg and HBV DNA in cell culture supernatant; Western blot was used to measure the level of HBV core protein in cells; reverse-transcription quantitative real-time PCR was used to measure the level of HBV RNA in cells; dual-luciferase reporter assay was used to observe the effect of cyclin D1 on the activity of HBV basic core promoter (BCP). GSE83148 dataset was used to investigate the correlation between CCND1 and HBV-related regulatory factors. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. Results The analysis of GSE84044 data showed that 7 cell cycle genes were significantly negatively correlated with HBV DNA load in liver tissue of the patients with HBV-related liver fibrosis (all r < -0.3, all P < 0.05), which included the CCND1 gene ( r =-0.474, P < 0.001). Exogenous expression of cyclin D1 and cyclin D1 T286A mutant reduced the levels of HBsAg, HBeAg, and HBV DNA in culture supernatant of the HBV replication cell model, as well as the levels of HBV core protein and HBV RNA in cells. Exogenous expression of cyclin D1 significantly inhibited the transcriptional activity of HBV BCP. The expression level of CCND1 in liver tissue of chronic hepatitis B patients was significantly positively correlated with the expression of APOBEC3G ( r =0.575, P < 0.001), SMC5 ( r =0.341, P < 0.001), and FOXM1 ( r =0.333, P < 0.001) which inhibited HBV replication, while it was significantly negatively correlated with the expression of the HBV entry receptor NTCP ( r =-0.511, P < 0.001) and HNF1α as the transcription factor for positive regulation of HBV replication ( r =-0.430, P < 0.001). Overexpression of cyclin D1 in HepG2 cells reduced the transcriptional levels of HNF1α and NTCP. Conclusion Cyclin D1 inhibits HBV transcription and replication possibly by downregulating the expression of HNF1α and NTCP.

Isoflurane anesthesia regulates transcription of RORα and Rev-erbα genes and their circadian rhythm changes in the hippocampus and cerebral cortex of mice / 中华神经医学杂志

Objective:To investigate the effects of isoflurane anesthesia on the transcription levels of clock genes RORα and Rev-erbα and their circadian rhythm changes in the hippocampus and cerebral cortex of mice. Methods:Thirty-six 7-week-old male C57BL/6J mice were divided into control group and isoflurane anesthesia group ( n=18) according to random number table method. They were subjected to 12 h light/12 h dark environment for 1 week. Mice in the isoflurane anesthesia group were anesthetised with 1.2% isoflurane at 22:00 pm on the night before the experiment for 6 h; 4 h after anesthesia, the light time of 8:00 am was taken as the zeitgeber time (ZT0), and then, every 8 h was taken as the recorded time (ZT8, ZT16, ZT24, ZT32 and ZT40). The RORα and Rev-erbα mRNA expressions were measured by real-time fluorescent quantitative PCR (RT-qPCR) at each time point in the cerebral cortex and hippocampus of the two groups. The cosine curves of RORα and Rev-erbα mRNA expressions were analyzed by Chronos-Fit software. Results:As compared with the control group, the isoflurane anesthesia group had significantly down-regulated RORα mRNA expression in the hippocampus, significantly up-regulated RORα mRNA expression in the cerebral cortex, and significantly down-regulated Rev-erbα mRNA expression in the cerebral cortex ( P<0.05). The RORα and Rev-erbα mRNA expressions in the hippocampus and cerebral cortex of the control group showed a rhythmic cycle of about 24 h; while the circadian rhythm of RORα mRNA shifted to the right by 5.41 h, and the circadian rhythm of Rev-erbα mRNA shifted to the left by 0.89 h in the hippocampus of the isoflurane anesthesia group. The peak circadian rhythm of RORα mRNA shifted to the right by 0.35 h, and the peak circadian rhythm of Rev-erbα mRNA shifted to the left by 0.56 h in the cerebral cortex of the isoflurane anesthesia group. Conclusion:Isoflurane anesthesia can induce the changes of RORα and Rev-erbα transcription levels and circadian rhythm changes in the hippocampus and cerebral cortex of mice.

Analysis of the endometrial transcriptome at the time of implantation in women receiving a single post-ovulatory dose of levonorgestrel or mifepristone

Abstract Background: Levonorgestrel (LNG) is a progesterone receptor agonist used in both regular and emergency hormonal contraception; however, its effects on the endometrium as a contraceptive remain widely unknown and under public debate. Objective: To analyze the effects of LNG or mifepristone (MFP), a progesterone receptor antagonist and also known as RU-486, administered at the time of follicle rupture (FR) on endometrial transcriptome during the receptive period of the menstrual cycle. Methods: Ten volunteers ovulatory women were studied during two menstrual cycles, a control cycle and a consecutively treated cycle; in this last case, women were randomly allocated to two groups of 5 women each, receiving one dose of LNG (1.5 mg) or MFP (50 mg) the day of the FR by ultrasound. Endometrial biopsies were taken 6 days after drug administration and prepared for microarray analysis. Results: Genomic functional analysis in the LNG-treated group showed as activated the bio-functions embryo implantation and decidualization, while these bio-functions in the T-MFP group were predicted as inhibited. Conclusions: The administration of LNG as a hormonal emergency contraceptive resulted in an endometrial gene expression profile associated with receptivity. These results agree on the concept that LNG does not affect endometrial receptivity and/or embryo implantation when used as an emergency contraceptive.

Consumption of minas frescal cheese may be a source of human infection by Campylobacter jejuni / Consumo de queijo minas frescal pode ser fonte de infecção humana por Campylobacter jejuni

Campylobacter spp. is an emerging pathogen that causes gastroenteritis in humans and the consumption of dairy food can characterize sources of infection. We aimed to verify the viability and a presence of transcripts associated with characteristics of virulence and adaptation of C. jejuni isolated from Minas Frescal cheeses, produced with contaminated milk and stored under refrigeration for up to ten days. The samples were analyzed for bioindicators, Campylobacter spp., pH, acidity, moisture and sodium chloride. Campylobacter spp. recovered were evaluated for the production of transcripts of: ciaB, dnaJ, p19 and sodB. The results were correlated with the viability of C. jejuni and changes in their transcriptome. Storage at lowtemperatures reduced C. jejuni from the first to the fourth day. The variations in humidity, pH and acidity influenced the decreasing of C. jejuni. There was a reduction in transcripts' production of the four genes, more pronounced on the fourth day, indicating the inability of the microorganism to perform its metabolic activities, due to the conditions of injury. Despite the presence of mechanisms of virulence and adaptation, C. jejuni could not remain viable four days after production. However, consumption of fresh cheese contaminated with Campylobacter jejuni can be a source of infection when consumed up to four days after production.

Campylobacter spp. é um patógeno emergente que causa gastroenterite em seres humanos e o consumo de produtos lácteos pode caracterizar fontes de infecção. O objetivo deste estudo foi verificar a viabilidade e a presença de transcritos associadas a características de virulência e adaptação de C. jejuniisoladas de queijos frescos, produzidos com leite contaminado e mantidos refrigeradas por dez dias. Foram analisados bioindicadores, Campylobacter spp., pH, acidez, umidade e cloreto de sódio. Campylobacter spp. recuperados foram avaliados quanto à produção dos transcritos: ciaB, dnaJ, p19 e sodB. Os resultados foram correlacionados com a viabilidade de C. jejuni e alterações no transcriptoma. O armazenamento em baixas temperaturas reduziu C. jejuni do primeiro ao quarto dia. As variações na umidade, pH e acidez influenciaram a queda de C. jejuni. Houve uma redução na produção de transcritos dos quatro genes, mais pronunciada no quarto dia, indicando a incapacidade do micro-organismo em realizar suas atividades metabólicas, devido às condições de injúria. Apesar da presença de mecanismos de virulência e adaptação, C. jejuni não permaneceu viável quatro dias após a produção. Porém, o consumo de queijo fresco contaminado com Campylobacter jejunipode ser uma fonte de infecção quando consumido até quatro dias após a produção.

Experimental Study on Inhibition Effect of Berberine in Escherichia coil Gene Transcription / 世界科学技术-中医药现代化

There have been many reports on berberine (BBR) effect of the inhibition on gut bacteria,but more from the protein level.In view of the preference of BBR for DNA binding,we here investigated the expression of BBR from the transcriptional expression level of the gene.The results showed that BBR had a higher affinity for UP element of Escherichia coli (E.coli) gene,and the transcription initiation region of this element contained TATA base sequence.The expression of genes sulA,recA and 16S which contain the genes of the UP element regulatory elements in the upstream of the promoter could be suppressed by BBR,and the expression of lpxC,secG and mutT which did not contain the genes of the UP element regulatory elements in the upstream of the promoter could not be inhibited by BBR.It is shown that the TATA sequence is the target of BBR.This result provides a new perspective for exploring the effect of BBR's inhibition of microbiota from gene transcription.

Berberine Effect of Anti-microbiota-Possibility and Clinical Significance on Gut Microbiota and Metabolism: A Review / 世界科学技术-中医药现代化

Berberine (BBR) is known as a classic drug for intestinal infection treatment.BBR inhibits intestinal bacteria,which is the core of its role in the treatment of intestinal infection.With the survival of local intestinal bacteria and its related metabolites on the physiological and pathological functions of the body continue to recognize the impact of it,more and more literatures have presented the effect of BBR through the impact of intestinal bacteria on the body glycol-lipid metabolism,even brain function.This allows us to re-understand the pathophysiology of BBR in inhibiting gut microbiome.In this paper,the antibacterial activity of BBR was reviewed and analyzed.The possible molecular target of BBR was analyzed according to the characteristics of prokaryotes gene expression,which was helpful to the in-depth study of BBR on intestinal bacteria.Thus,a more comprehensive understanding of the pharmacological effects of BBR is given.

Principales promotores utilizados en la transformación genética de plantas / Main promoters used in plant genetic transformation

El conocimiento pleno de los promotores determina el éxito en la obtención de nuevos cultivares de plantas a través de técnicas biotecnológicas, ya que dicha secuencia del ADN regula la transcripción de otras regiones adyacentes o cercanas, encontrándose los siguientes promotores: constitutivos, tejido-específicos o estadio-específicos, inducibles y sintéticos. En esta revisión se resume de manera precisa los conceptos, ventajas y limitaciones de los distintos tipos de promotores, con ejemplos claros de ello.

Full knowledge of promoters determines success in obtaining new plant cultivars through biotechnology techniques. This DNA sequence regulates the transcription of adjacent or nearby regions, which are mainly constitutive, tissue-specific or stage-specific, inducible and synthetic. This review summarizes the precise concepts, advantages and limitations of different types of promoters, including clear examples of them.

Immunohistochemical expression of Ets‑related gene‑transcriptional factor in adenocarcinoma prostate and its correlation with Gleason score.

Background: Prostate carcinoma is the second leading cause of cancer‑related deaths in males worldwide. The burden is expected to grow 1.7 million new cases and 499,000 new deaths by 2030. In developing countries such as India, prostate carcinoma will show an increase by 140% in the next few years. Although the diagnosis of prostate carcinoma can usually be made on histological features, now a days many immunohistochemical (IHC) markers are used to distinguish it from benign mimickers as well as in predicting prognosis and treatment. Out of these markers, Ets‑related gene (ERG product) is a proto‑oncogene which participates in chromosomal translocations and is frequently over expressed in prostate carcinoma which harbors ERG‑transmembrane protease, serine 2 fusion. Materials and Methods: Fifty cases of carcinoma prostate diagnosed in needle biopsies and prostatic chips, in the Department of Pathology of a tertiary care teaching hospital in Punjab, India, were included in the present study. The slides were observed under the light microscope, and Gleason scoring was done using the 2005 International Society of Urological Pathology modified Gleason system. IHC study for ERG expression was done on all the cases, for which anti‑ERG monoclonal rabbit clone antibody EP111 (Dako, Denmark) was used. Lymphocytes and endothelial cells were taken as in built positive controls for staining. The intensity of ERG positivity was scored as no staining (0), weak staining (+1), moderate staining (+2) and intense staining (+3). The H score was then calculated by multiplying the intensity of the stain with the percentage (0–100) of the cells showing that staining intensity. The H‑score has a range of 0–300. The relationship between IHC expression and clinico‑pathological parameters was compared and analyzed using Chi-square test. P < 0.05 was considered statistically significant. Results: Majority of patients included in the study were in the age group of 61–80 (84% of the total). When ERG expression was studied with age‑specific rates, it was not found to be statistically significant. The most common pattern noted in the present study was 4 + 3, constituting 36% of total, followed by 3 + 4 constituting 32%. Calculating the score, the majority of patients had a Gleason score of 5–8, constituting 76% of total. Out of the total fifty cases of prostate carcinoma, ERG was positive in 29 cases (58%) and negative in 21 cases (42%). Fourteen out of 21 (48%) of the ERG positive cases had an intensity score of 3. When the ERG intensity was correlated with the Gleason score group, it was seen that patients having Gleason score 7–8 showed ERG positivity in 19 out of 38 cases (50%), with 11/19 (57%) cases showing an ERG intensity score of 3. The Gleason score group 9–10 showed ERG positivity in 83% (10/12) cases, 20% (2/10) cases showing intensity score of 3. This correlation was found to be statistically significant. Conclusion: ERG immunostaining was performed in a small Indian cohort of prostate cancer patients, diagnosed in trucut biopsy specimens and prostatic chips. ERG expression was found in 58% patients. An increase in the ERG expression was observed with an increase in Gleason score. The intensity of ERG expression, however, decreased with an increasing Gleason score.

Chronopharmacokinetics of gefitinib and its mechanisms in tumor-bearing nude mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of the dosing time on the pharmacokinetics of gefitinib and its potential mechanism. METHODS Female BALB/c nude mice were housed under standardized 12 h light/dark circadian conditions(light on at 7:00,off at 19:00)for two weeks before a non-small cell lung cancer(NSCLC)model was established. Two weeks later,they were divided into 2 groups(8:00,20:00)randomly. Gefitinib was orally administered at the dose of 1 mg · kg-1 to the mice in each group at 8:00 or 20:00,respectively. Blood was collected at 10 different time points after each administration. Livers were collected every 4 h during the 24 h period from the non-administrated nude mice of NSCLC model. The plasma concentration of gefitinib was determined through an HPLC-MS/MS and the parameters were calculated by WinNonlin 6.3. The total RNA was extracted from livers,purified, synthesized to cDNA that was subjected to qRT-PCR analysis for mRNA expression levels of cyto?chrome P450 enzymes(Cyp)3a11,Cyp3a13,pregnane X receptor(PXR)and constitutive androstane receptor (CAR). RESULTS The area under the plasma concentration-time curve (AUC) and mean residence time (MRT) of 8:00 administration group were higher than those of 20:00 administration group(P<0.05). The clearance(Clz/F) of 8:00 administration group was lower than that of 20:00 administration group(P<0.05). The mRNA expression levels of PXR and CAR were consistent with those of Cyp3a11 and Cyp3a13. CONCLUSION Circadian rhythm exists in the pharmacokinetics of gefitinib and it may be closely related to CYP3A and its regulator genes.

The role of rs10993994 for the transcriptional activity of MSMB gene in prostate cancer cell lines / 中国癌症杂志

Background and purpose:Rs10993994 at 10q11 was found to be associated with prostate cancer risk by genome-wide association studies. This study tried to reveal its mechanism and to explore the role of rs10993994 for the transcriptional activity of microseminoprotein beta (MSMB) gene in prostate cancer cell lines. Methods:Promoter fragments were generated by chemical synthesis. Due to the two possibility of rs10993994 (T/C) in the region, we generated two promoter fragments: MSMB promoter-T and MSMB promoter-C. The fragments were then cloned into pGL3-basic vectors, positive clones were transfected into prostate cancer cell lines PC-3 and LNCaP, ifnally, relative level of lfuorescence was detected by lfuoresce detector.Results:We generated two promoter fragments of MSMB, MSMB promoter-T and MSMB promoter-C. The two promoter fragments were cloned with pGL3-basic vectors to pGL3-MSMB promoter-T and pGL3-MSMB promoter-C. In PC-3, the relative level of lfuorescence of pGL3-MSMB promoter-C was signiifcant higher than that of pGL3-MSMB promoter-T(2.27±0.39vs 0.57±0.13,P<0.05); In LNCaP, the relative level of lfuorescence of pGL3-MSMB promoter-C was signiifcant higher than that of pGL3-MSMB promoter-T (1.70±0.32vs 0.37±0.09,P<0.05).Conclusion:The transcriptional activity of pGL3-MSMB promoter-C was stronger than that of pGL3-MSMB promoter-T. Rs10993994 could inlfuence the transcriptional activity ofMSMB gene promoter in prostate cancer cell lines PC-3 LNCaP, allele C in rs10993994 could facilitate transcription than T.

Las adicciones, la genómica y la proteómica / Addictions, genomics and proteomics

Drug addiction is a chronically relapsing disorder that has been characterized by (1) compulsion to seek and take the drug, (2) loss of control in limiting intake, and (3) emergence of a negative emotional state (e.g, dysphoria, anxiety, irritability) reflecting a motivational withdrawal syndrome when access to the drug is prevented (defined as Substance Dependence by the Diagnostic and Statistical Manual of Mental Disorders [DSM] of the American Psychiatric Association). Acute exposure to drugs of abuse initiates molecular and cellular alterations in the Central Nervous System that lead to an increased overall vulnerability to addiction with subsequent drug exposures. These drug-induced alterations employ changes in gene transcription that result in the synthesis of new proteins. Therefore, one of the important goals of addiction research is to identify the drug-induced gene expression changes in the specific brain structures related to the addictive properties of various drugs. The molecular and genomic mechanisms by which drugs of abuse induce neuroplastic changes related to addiction remain largely unknown. Several studies have evaluated changes in gene and protein expression profiles in the brain after administration of drugs of abuse. Exposure to psychostimulants induces the activity-dependent gene expression of several transcription activators and repressors. Genomic research strategies have recently transitioned from the search for unknown genes to the identification and evaluation of coordinated gene networks and transcriptional signatures. New opportunities arising from the analysis of these networks include identifying novel relationships between genes and signaling pathways, connecting biological processes with the regulation of gene transcription, and associating genes and gene expression with diseases. The identification of gene networks requires large gene expression data sets with multiple data points. Functional genomics methods, studying the steady-state levels of these mRNA species, such as quantitative RT-PCR (qRT-PCR), whole-genome microarray analysis, and next generation sequencing methods, provide sensitive and high-throughput approaches to quantitatively examining mRNA (and miRNA) species present within the cells of the Nervous System. Functional genomics studies can help to illuminate genes involved in the development of behaviors related to drug abuse and relapse liability, but cannot provide insight into post-translational modifications (e.g., phosphorylation and glycosylation of proteins after translation has occurred) or subcellular localization of the protein product. Therefore, using proteomic techniques presents the opportunity to assess the totality of gene expression, translation, modification, and localization. Unfortunately, the sensitivity of proteomic tools lags behind those of functional genomics. Moreover, examining the mRNA provides a restricted view of primarily the cell body. Indeed, from a systems biology standpoint, analysis of both mRNA and protein levels (as well as miRNA and epigenetic changes) will ultimately provide a more integrated view of the molecular underpinnings of addiction. When applying proteomic technologies to addiction research, an understanding of the power of proteomic analysis is essential. After genetic information is transcribed into mRNA, a template is provided to the cell from which proteins will be synthesized. Neuroproteomic studies offer great promise for increasing understanding of the biochemical basis of addiction. While proteomics is still an evolving field, proteomic approaches have proven useful for elucidating the molecular effects of several drugs of abuse. With a number of ongoing research programs in addiction proteomics and a growing number of investigators taking advantage of these tools, the addiction research field will benefit from a consideration of the capabilities and limitations of proteomic studies. As with other biomedical research fields, drug abuse research is making use of new proteomic capabilities to examine changes in protein expression and modification on a large scale. To obtain the maximum benefit and scientific advancement from these new technologies, a clear understanding of the power and limitations of neuroproteomics is necessary. With the main limitation of neuroproteomic studies being the complexity of the proteome, approaches that focus these studies need to be employed. The salient message is that there is not a single best technical approach for all studies and that the main driver for the choice of proteomic technology and experimental design should be the advancement of the understanding and treatment of drug abuse. An important area that has heretofore received limited attention is the experimental design and interpretation specific to neuro-proteomic studies of drug abuse. These challenges include choice of animal model, ensuring sample quality, the complexity of brain tissue, confirming discovery findings, data analysis strategies, and integration of large data sets with the existing literature. Epigenetics is the study of heritable changes other than those in the DNA sequence and encompasses two major modifications of DNA or chromatin: DNA methylation and post-translational modification of histones. In this context, now it is known that regulation of gene expression contribute to the long-term adaptations underlying the effects of drugs of abuse. The precise molecular events that are required for modification of chromatin and that underlie gene repression or activation have not been elucidated. Recent reports have addressed this question and demonstrated that drugs of abuse modify specific methyl-CpG-binding proteins that control histone acetylation and gene expression. Further elucidation of the wide-range of histone modifications and the ensuing consequences on gene expression will be necessarily before the potential for drug development can be realized. It is important to characterize the molecular alterations underlying chromatin remodeling and the regulation of the epigenetics events by drugs of abuse. It is clear that modification in gene expression by drugs of abuse promote cellular changes. This review is intended to provide guidance on recent advances in the field of drug addiction. This review also presents a number of experimental design and sample approaches that have been applied to genomic, proteomic and epigenetic studies of addiction. Coupled with new technologies for data collection, analysis, and reporting, these approaches represent the future of the addiction field and hold the key to unlocking the complex of profile of drug abuse disorders.

La adicción a las drogas es una enfermedad mental que se caracteriza por ocasionar graves implicaciones sociales, económicas y de salud de los individuos que la padecen. La exposición aguda a las drogas de abuso provoca alteraciones moleculares y celulares en el Sistema Nervioso Central que ocasionan una vulnerabilidad para sufrir adicción a subsecuentes exposiciones a sustancias de abuso diferentes. Las alteraciones inducidas por las drogas producen cambios en la transcripción de genes que resultan en la síntesis de nuevas proteínas. Uno de los objetivos importantes en la investigación en el campo de las adicciones es identificar los cambios en la expresión de genes inducidos por las drogas en estructuras específicas del cerebro que están relacionadas con las propiedades adictivas de diferentes sustancias. El campo de la genómica y la proteómica, aplicada al estudio de las adicciones, tiene como objetivo identificar a los genes y las proteínas candidatos involucrados en la regulación de los procesos adictivos. Se han logrado progresos considerables en la identificación de genes y proteínas que regulan las conductas complejas presentes en los procesos adictivos en modelos de animales y modelos de estudio en humanos con material obtenido post-mortem. Estos descubrimientos se han sumado a los esfuerzos por identificar los circuitos neurales implicados en las manifestaciones conductuales relacionadas con las adicciones. También han permitido la identificación de genes candidatos que podrán ser blancos de futuras estrategias terapéuticas desarrolladas para tratar los procesos adictivos. Los estudios de genómica funcional han permitido identificar algunos de los genes involucrados en el desarrollo de las conductas adictivas, pero no tienen la capacidad de proporcionar información sobre las modificaciones post-traduccionales ni de la localización sub-celular de las proteínas para las que codifican los genes. Por lo tanto, la incorporación de estudios proteómicos ofrece la oportunidad de lograr evaluar, en su totalidad, la expresión, la traducción, las modificaciones y la localización de los genes y sus productos de expresión. Para obtener los máximos beneficios y avances con el empleo de estas nuevas tecnologías, deben comprenderse en su totalidad los alcances y limitaciones de la neuroproteómica. En este sentido, se debe tener especial cuidado en la elección del modelo de estudio, asegurar la calidad de la muestra, la complejidad de la estructura en estudio, confirmar los resultados obtenidos, las estrategias de análisis de resultados y la integración de los datos obtenidos con los ya reportados en la literatura científica. Los estudios recientes sobre los mecanismos moleculares que controlan los cambios inducidos por las drogas de abuso sobre la función transcipcional, la conducta y la plasticidad sináptica han identificado el importante papel que desempeña la remodelación de cromatina en la regulación y estabilidad de los programas genéticos neuronales mediados por las drogas y la subsecuente manifestación de las conductas adictivas. Se han identificado alteraciones epigenéticas sobre el genoma, tales como metilación del DNA y modificaciones en la función de las proteínas histonas. Estos importantes mecanismos se ven afectados como una respuesta neurobiológica a la administración de sustancias de abuso. Esta revisión pretende mostrar algunos de los avances recientes en el campo de las adicciones, presentando una breve descripción de los hallazgos que emplean aproximaciones genómicas, proteómicas y epigenéticas. Las implicaciones de estos estudios moleculares ponen de manifiesto nuevos conocimientos sobre el probable desarrollo de intervenciones terapéuticas en el futuro.

Transcript atlas of blood coagulation-associated gene uncovers physiological activities in 8 liver cell types of rat regenerating liver / 解剖学报

ObjectiveTo study the transcript atlas of cell immunity-associated genes in 8 liver cell types including hepatocytes, bile duct epithelial cells, oval cells, hepatic stellate cells, sinus endothelial cells, Kupffer cells, pit cells and dendritic cells from the rat regenerating liver. Methods Isolation of rat 8 liver cell types, detection of their genes expression change and the prediction physiological activities were accorded to cluster program, and the methods of bioinformation and systematic biology, and their gene expression patterns were analyzed by Microsoft Excel software. Results A total of 40 blood coagulation-associated genes yielded the meaningful expression changes in liver regeneration. Serpine1, a2m in eight liver cell types, vwf, klkb1 in seven liver cell types other than Kupffer cells, and other genes in the two or more liver cell types yielded the meaningful expression changes. It suggested that the synthesis of kallikrein and prothrombin、the formation of thrombin were enhanced at the priming and progressing phases of liver regeneration. The biological activities, such as fibrin monomer aggregated into fibrin polymers, were elevated at termination phase.Conclusion The rat liver regeneration is closely associated with the blood coagulation.

Gene transcript profiles of liver cells predict that AI408225 promotes antigen peptide combining with TCR in rat regenerating liver / 解剖学报

ObjectiveTo explore the correlation of unknown gene AI408225 and humoral immunity involved in rat regenerating liver. MethodsIsolation of rat 8 liver cell types, detection of their genes expression change were accorded to Percoll density gradient centrifugation combined with immune magnetic beads separation method, and their genes sequence homoeology and coexpression relation were analyzed by Microsoft Excel, BLAST software, and the prediction physiological activities were analyzed by bioinformatics and systematic biology. Results A total of 93 humoral immunity-ass ociated genes yielded the meaningful expression changes in liver regeneration, the corresponding gene numbers in 8 liver cell types included hepatocytes, bile duct epithelial cells, oval cells, hepatic stellate cells, sinus endothelial cells, Kupffer cells, pit cells and dendritic cells were 33, 46, 17, 59, 48, 35, 53, 68. Among that AI408225 and cd4 are homology and coexpression.Conclusion The rat liver regeneration is closely associated with humoral immunity, and AI408225 participates in the formation of compound of MHC II-antigen peptide-CD4-TCR.

Transcript atlas of cell immunity-associated gene uncovers physiological activities in 8 liver cell types of rat regenerating liver / 解剖学报

ObjectiveTo study the transcript atlas of cell immunity-associated genes in 8 liver cell types including hepatocytes, bile duct epithelial cells, oval cells, hepatic stellate cells, sinus endothelial cells, Kupffer cells, pit cells and dendritic cells from the rat regenerating liver. Methods The 8 liver cell types were isolated respectively by Percoll density gradient centrifugation combined with immune magnetic beads separation method. Their expression changes in the regenerating livers were detected by Rat Genome 230 2.0 Array, the expression patterns and the predicted physiological activities of cell immunity-associated genes were analyzed by Cluster program, and the methods of bioinformation and systematic biology. Results A total of 40 cell immunity-associated genes yielded the meaningful expression changes in liver regeneration, the corresponding gene numbers in 8 liver cell types were 19, 19, 9, 19, 19, 21, 22 and 21, respectively. It suggested that the formation of antigen peptide-MHC complexes, the NF-κB kinase activity and the production of cytokines like IL-2 were enhanced at the priming and progressing phases of liver regeneration. The biological activities, such as NF-κB-promoting cell differentiation and caspase-induced T cell apoptosis, were elevated at termination phase.Conclusion The rat liver regeneration is associated with cell immunity.

Transcriptome Analysis of Artemisinin Biosynthetic Genes / 广州中医药大学学报

Objective To investigate the expression patterns of artemisinin biosynthetic genes in Artemisia annua L.during the development stage and in different tissues,and to explore the mechanism of spatial and temporal modulators for artmisinin production.Methods The transcriptional profiles of artemisinin biosynthetic genes in the capital and cooperative pathways were quantitatively assayed by real time fluorescence quantitative-polymerase chain reaction(RTFQ-PCR) in different tissues of roots,stems,leaves and flowers,and in leaf-flourishing,pre-floral,and post-floral periods.Results The expression levels of the tested genes were extremely low in June,raised in July,and reached their peak values in August(before flowering),but dropped gradually in September(after blooming).In August,the transcription levels of the tested genes increased by 3 to 15 times compared to the lowestlevels.In particular,ADS and CYP71AV1 mRNA levels had the great elevation,which were 12 and 15 times as much as those in June.During the flowering period,the artemisinin biosynthetic genes mRNA expression was detectable in roots,stems,leaves and flowers,and the expression levels had no obvious difference except that ADS mRNA level in leaves was 2 times higher than that in other tissues(P

Human p100 protein:a novel coactivator that enhances ST AT6-mediated gene transcriptional activation / 中国免疫学杂志

Objective:To setup a stable cell line which can overexpress human p100 protein,and then st udy the physiologic functions of p100 in regulation of STAT6-mediated gene tran scription.Methods:Using coimmunoprecipitation method to identify protein-protein interactions.STA T6-mediated gene transcriptional activation was detected by luciferase assay.Results:Human p100 protein interacted with both STAT6 and RNA polymerase Ⅱ,resulting in the enhancement of STAT6-mediated gene transcriptional activation.Conclusion:Human p100 protein acts as a bridging factor between STAT6 and basal transcripti onal machenary. [

Chinese herbs improve transcription of the gene for albumin in nephrotic syndrome rats / 中华肾脏病杂志

; 3. therapy with A&A (A&A); 4. Astragali alone (A); 5. Astragali polysac-charide Ⅰ (APⅠ); 6. AP Ⅱ ; 7. Astragali glucoside (AG), Results The level of serum albumin,albumin mRNA and albumin gene transcription were measured by biochemistry, Northern blot hybridization, nuclear run-on assay and quantity in laser density screening. The level of serum albumin in N was significantly lower than C. The serum albumin concentration in each therapy group was higher than N group. The transcription of the albumin gene was higher in N than in C and highest in A&A. The alterations of northern blot hybridization were same as the transcription results. But both the level of albumin mRNA and the transcription of albumin gene in A, AP I , AP I and AG were no change compared with N. Conclusion A&A increases albumin mRNA expression at least in part by improving the rate of gene transcription, which participate the protection of the decrease of serum albumin in NS. But the effect of A, AP 1 , AP I and AG may mediated by other unknown mechanisms.

Effect of sibelium on apoptosis and transcription of converting gene in limbic system of rats' cerebral ischemia / 临床神经病学杂志

Objective To study the effect and the mechanism of sibelium on apoptosis and transcription of bcl 2 clan and interleukin 1? converting enzyme(ICE) gene in limbic system after rats' middle cerebral artery occlusion(MCAO).Methods MCAO model was made, and determined the dynamic changes on the cell counts of apoptosis, bcl 2 clan and ICE gene positive transcription in temple ortex, hippocampus and hypothalamus in the controls of normal saline,sibelium 50 ?g intervenient group and sibelium 100 ?g intervenient group during rats MCAO or reperfusion.Results The apoptosis counts of every times in sibelium 100?g intervenient group was significant lower than another two groups( P 0.05).The bcl 2 gene positive transcription cell counts at every times in sibelium 100?g intervenient group were significant higher than another two gruops( P

REGULATION OF EXOGENOUS POLYAMINES ON ORNITHINE DECARBOXYLASE GENE TRANSCRIPTION IN REGENERATING RAT HEPATOCYTES / 解剖学报

Objective To study the regulation of natural polyamines(putrescine,spermidine and spermine) on ornithine decarboxylase gene transcription in regenerating rat hepatocytes and to assess the roles of polyamines in liver regeneration. Methods Rat regenerating liver was induced by surgical partial hepatectomy(PH).The expressive levels of ODC mRNA in regenerating liver were measured by in situ hybridization analysis.Exogenous polyamines(dissolved in 0.9% NaCl) were administered subcutaneously to adult male Sprague-Dawley rats. Results ODC mRNA expression was always lower in high dose(20 mg/kg body weight) putrescine-treated rats compared with that in control rats during the observed regeneration,especially at the 2(nd),4(th) and 10(th) hours(P