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Developing a common medication strategy for disease control and management could be greatly beneficial. Investigating the differences between diseased and healthy states using differentially expressed genes aids in understanding disease pathophysiology and enables the exploration of protein-drug interactions. This study aimed to find the most common genes in diarrhea-causing bacteria such as Salmonella enterica serovar Typhimurium, Campylobacter jejuni, Escherichia coli, Shigella dysenteriae (CESS) to find new drugs. Thus, differential gene expression datasets of CESS were screened through computational algorithms and programming. Subsequently, hub and common genes were prioritized from the analysis of extensive protein-protein interactions. Binding predictions were performed to identify the common potential therapeutic targets of CESS. We identified a total of 827 dysregulated genes that are highly linked to CESS. Notably, no common gene interaction was found among all CESS bacteria, but we identified 3 common genes in both Salmonella-Escherichia and Escherichia-Campylobacter infections. Later, out of 73 protein complexes, molecular simulations confirmed 5 therapeutic candidates from the CESS. We have developed a new pipeline for identifying therapeutic targets for a common medication strategy against CESS. However, further wet-lab validation is needed to confirm their effectiveness., (© 2022. The Author(s).)

Insufficient invasion of trophoblast cells into the uterine decidua is associated with preeclampsia (PE). G protein-coupled estrogen receptor (GPER) is a membrane estrogen receptor involved in non-genomic estrogen signaling. GPER is expressed in human trophoblast cells and downregulated GPER levels are noted in PE. However, to date, the role of GPER in trophoblast cells remains largely unknown. Here, we applied RNA sequencing (RNA-seq) to HTR-8/SVneo human trophoblast cells in response to G1, an agonist of GPER, and identified angiopoietin-like 4 (ANGPTL4) as a target gene of GPER. Treatment of trophoblast cells with G1 or 17β-estradiol (E2) activated Yes-associated protein (YAP), the major downstream effector of the Hippo pathway, via GPER but in a mammalian STE20-like protein kinase 1 (MST1)-independent manner. Using pharmacological inhibitors as well as loss- and gain-of-function approaches, our results revealed that YAP activation was required for GPER-stimulated ANGPTL4 expression. Transwell invasion assays demonstrated that activation of GPER-induced ANGPTL4 promoted cell invasion. In addition, the expression levels of GPER, YAP, and ANGPTL4 were downregulated in the placenta of patients with PE. Our findings reveal a mechanism by which GPER exerts its stimulatory effect on human trophoblast cell invasion by upregulating YAP-mediated ANGPTL4 expression., (© 2021. The Author(s).)

Bacterial disease outbreaks in filter feeder bivalve Hyriopsis cumingii as water contamination become more frequent in the water ecosystem, especially in intensive aquaculture habitats. To characterize host-pathogen interactions between H. cumingii and bacterial infection, we investigated the effects of Stenotrophomonas maltophilia HOP3 and Aeromonas veronii GL1 on the antioxidant response, tissue invasion and transcriptome expression of H. cumingii by infectivity trials. We showed that bacterial infections resulted in tubular necrosis of the hepatopancreas and induced the acute immune response in H. cumingii. The transcriptomic study identified a total of 5957 differentially expressed genes (DEGs) after A. veronii challenge. These DEGs were implicated in 302 KEGG pathways, notably in Apoptosis, Phagosome and Lysosome. The results showed that the relative expressions of all six immune-related DEGs were effectively stimulated with A. veronii, accompanied by tissue differences. Overall, these findings will contribute to an analysis of the immune response of H. cumingii to bacterial infection at the transcriptomic level and its genomic resource for research., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Heat shock proteins (HSPs) represent a group of important proteins which are produced by all kinds of organisms especially under stressful conditions. DnaK, an Hsp70 homolog in prokaryotes, has indispensable roles when microbes was confronted with stress conditions. However, few data on DnaK from Rhodococcus sp. were available in the literature. In a previous study, we reported that toluene and phenol stress gave rise to a 29.87-fold and 3.93-fold increase for the expression of DnaK from R. ruber SD3, respectively. Thus, we deduced DnaK was in correlation with the organic solvent tolerance of R. ruber SD3., Objective: To elucidate the role of DnaK in the organic solvent tolerance of R. ruber SD3, expression, purification and functional analysis of Dnak from R. ruber SD3 were performed in the present paper., Methods: In this article, DnaK from R. ruber SD3 was heterologously expressed in E. coli BL21(DE3) and purified by affinity chromatography. Functional analysis of DnaK was performed using determination of kinetics, docking, assay of chaperone activity and microbial growth., Results: The recombinant DnaK was rapidly purified by affinity chromatography with the purification fold of 1.9 and the recovery rate of 57.9%. Km, Vmax and Kcat for Dnak from R. ruber SD3 were 80.8 μM, 58.1 nmol/min and 374.3 S -1 , respectively. The recombinant protein formed trimer in vitro, with the calculated molecular weight of 214 kDa. According to in-silico analysis, DnaK interacted with other molecular chaperones and some important proteins in the metabolism. The specific activity of catalase in the presence of recombinant DnaK was 1.85 times or 2.00 times that in the presence of BSA or Tris-HCl buffer after exposure to 54 °C for 1h. E. coli transformant with pET28-dnak showed higher growth than E. coli transformant with pET28 at 43°C and in the presence of phenol, respectively., Conclusion: The biochemical properties and the interaction analysis of DnaK from R. ruber SD3 deepened our understanding of DnaK function. DnaK played an important role in microbial growth when R. ruber was subjected to various stress such as heating and organic solvent., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The aim of this study is to investigate the therapeutic role of Tanshinone II A, a key integrant from salvia miltiorrhiza, against pathological vascular remodeling. Completed ligation of mouse left common carotid arteries animal model and rat smooth muscle cells used to investigate the role of Tanshinone II A in regulating pathological vascular remodeling through hematoxylin and eosin staining, immunohistochemistry staining, immunofluorescence staining, adenovirus infection, real time PCR and western blotting. Our data demonstrated that Tanshinone II A treatment suppresses vascular injury-induced neointima formation. In vitro studies on rat smooth muscle cell indicated that Tanshinone II A treatment attenuates PDGF-BB induced cell growth, and promotes smooth muscle cell differentiated marker genes expression that induced by rapamycin treatment. Tanshinone II A treatment significant inhibits rat smooth muscle cell proliferation and migration. Tanshinone II A promotes KLF4 expression during smooth muscle phenotypic switching. Overexpression of KLF4 exacerbates Tanshinone II A mediated smooth muscle cell growth inhibition. Tanshinone II A plays a pivotal role in regulating pathological vascular remodeling through KLF4 mediated smooth muscle cell phenotypic switching. This study demonstrated that Tanshinone II A is a potential therapeutic agent for vascular diseases.

Genetic factors do not fully account for the relatively high heritability of neurodevelopmental conditions, suggesting that non-genetic heritable factors contribute to their etiology. To evaluate the potential contribution of aberrant thyroid hormone status to the epigenetic inheritance of neurological phenotypes, we examined genetically normal F2 generation descendants of mice that were developmentally overexposed to thyroid hormone due to a Dio3 mutation. Hypothalamic gene expression profiling in postnatal day 15 F2 descendants on the paternal lineage of ancestral male and female T3-overexposed mice revealed, respectively, 1089 and 1549 differentially expressed genes. A large number of them, 675 genes, were common to both sets, suggesting comparable epigenetic effects of thyroid hormone on both the male and female ancestral germ lines. Oligodendrocyte- and neuron-specific genes were strongly overrepresented among genes showing, respectively, increased and decreased expression. Altered gene expression extended to other brain regions and was associated in adulthood with decreased anxiety-like behavior, increased marble burying and reduced physical activity. The sperm of T3-overexposed male ancestors revealed significant hypomethylation of CpG islands associated with the promoters of genes involved in the early development of the central nervous system. Some of them were candidates for neurodevelopmental disorders in humans including Nrg3, Nrxn1, Gabrb3, Gabra5, Apba2, Grik3, Reln, Nsd1, Pcdh8, En1, and Elavl2. Thus, developmental levels of thyroid hormone influence the epigenetic information of the germ line, disproportionately affecting genes with critical roles in early brain development, and leading in future generations to disease-relevant alterations in postnatal brain gene expression and adult behavior.

Olive flounder (Paralichthys olivaceus) is a commercially important flatfish species cultured in East Asia. Female flounders generally grow more rapidly than males, therefore control of the sex ratio seems to be a proposed way to increase production. However, the sex determination gene and sex determination mechanism have yet been elucidated. The brain is an important organ that is involved in gonadal development. To explore the sex differences of gene expression in the brain before and during the flounder gonadal differentiation, we used messenger RNA (mRNA)-seq technology to investigate transcriptomes of male and female brains. Between female and male brains, 103 genes were differentially expressed before ovarian differentiation, 16 genes were differentially expressed before testicular differentiation, and 64 genes were differentially expressed during gonadal differentiation. According to annotation and Kyoto Encyclopedia of Genes and Genomes information, the differentially expressed genes (DEGs) were involved in circadian rhythm, circadian rhythm-fly, circadian entrainment, dopaminergic synapse, calcium signaling, glutamatergic synapse, taste transduction, herpes simplex infection, long-term depression, retrograde endocannabinoid signaling, and the synaptic vesicle cycle pathways. MicroRNA (miRNA)-seq was performed during the gonadal differentiation and the target genes of miRNAs were predicted. Integrated analysis of mRNA-seq and miRNA-seq showed that 29 of the 64 DEGs were regulated by the differentially expressed miRNAs during the gonadal differentiation. Our study provides a basis for further studies of brain sex differentiation and the molecular mechanism of sex determination in olive flounder., (© 2019 Wiley Periodicals, Inc.)

Objective: Programmed death-ligand-1 (PDL1) is a molecule involved in immune evasion in various kinds of tumors. Here, we aim to determine whether the expression of PDL1 protein is related to the response of patients to neoadjuvant therapy and survival outcome., Methods: Immunohistochemistry (IHC) was performed on paraffin-embedded tumor samples from core needle biopsy before neoadjuvant therapy (NAT). Univariate and multivariate logistic regression were used to analyze the associations between PDL1 protein expression and pathological complete response (pCR) outcome. Kaplan-Meier plot and log-rank test were used to compare disease-free survival (DFS) between groups. A cox proportional hazards model was used to calculate the adjusted hazard ratio (HR) with 95% confidential interval (95%CI)., Results: A total of 94 patients were included for IHC testing. PDL1 protein expression on tumor cells was associated with better pCR rate in both univariate (OR = 2.621, p = 0.043) and multivariate (OR = 3.595, p = 0.029) logistic regression analysis. It was also associated with shorter DFS both by log-rank test (p = 0.015) and cox hazard model (HR = 22.824, 95%CI 1.621-321.284, p = 0.020). In hormone receptor (HR)-positive patients, PDL1 protein expression was also associated with better pCR (OR = 2.362, p = 0.022). It was also associated with poor DFS (HR = 18.821, 95%CI 1.645-215.330, p = 0.018)., Conclusions: Our results show that PDL1 protein expression is a predictive biomarker of pCR and a prognostic factor of DFS in breast cancer patients and HR-positive subgroups.

Gestational diabetes mellitus (GDM) represents a common carbohydrate metabolism disorder during pregnancy. The objective of this study was to evaluate the expression levels of mitofusin 2 (MFN2) expression in placentae of GDM patients compared to that in the placental tissues from normal uncomplicated pregnancies. A total of 70 subjects were enrolled from September 2014 to June 2016, including 42 patients with GDM (the GDM group) and 28 normal uncomplicated pregnancies (the control group). Immunohistochemical staining and qRT-PCR were used for the detection of the expression levels and distribution of MFN2 in the placentae of GDM patients and normal controls. Kolmogorov-Smirnov test was used for statistical analysis. P < 0.05 and P < 0.01 were used for assessing statistical significance. The baseline characteristics were comparable in both groups. The 1-h and 2-h postprandial glucose levels (PPG) were 7.94±1.26 versus 6.88±0.51 mmol/L and 7.01±1.34 versus 6.14±0.63 mmol/L, respectively,for the GDM group and the control group ( P < 0.05). The relative expression levels of MFN2 mRNA were 0.982±1.242 for GDM and 1.257 ± 0.815 for control, respectively, with significant between group difference ( P < 0.01). Immunohistochemical staining analysis showed that MFN2 was mostly distributed in the cytoplasm of syncytiotrophoblasts under optical microscopy. Additionally,about 50% of samples of the GDM group were within the intensity of moderate staining of MFN2 and more than 50% of patients in the control group were within the intensity of strong staining of MFN2. The expression levels of MFN2 in GDM placentae was significantly lower compared to that of placentae from normal uncomplicated pregnancies.

Background: Real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a critical tool for evaluating the levels of mRNA transcribed from genes. Reliable RT-qPCR results largely depend on normalization to suitable reference genes. Middle cerebral artery occlusion (MCAO) and oxygen-glucose deprivation/reoxygenation (OGD/R) are models that are commonly used to study ischemic stroke. However, the proper reference genes for RNA analysis in these two models have not yet been determined., Results: In this study, we evaluated the expression levels of six candidate housekeeping genes and selected the most suitable reference genes for RT-qPCR analyses of the cortices of MCAO mice and OGD/R-injured N2a cells. Four software programs, geNorm, NormFinder, BestKeeper and RefFinder, were used to validate the stabilities of the candidate reference genes. The results revealed that HPRT and 18S were the most stable reference genes in the cortices of MCAO mice and that β-actin and cyclophilin were the most stable reference genes in the OGD/R-injured N2a cells; in contrast, GAPDH and Sdha were the least stable genes in the cortices of MCAO mice and the OGD/R-injured N2a cells, respectively. Moreover, a combination of HPRT, 18S and cyclophilin was most suitable for normalization in analyses of the cortices of MCAO mice, and a combination of β-actin, cyclophilin, GAPDH, and 18S was most suitable for analyses of the OGD/R-injured N2a cells., Conclusions: This study provides appropriate reference genes for further RT-qPCR analyses of in vivo and in vitro ischemic stroke and demonstrates the necessity of validating reference genes for RNA analyses under variable conditions.

High-producing dairy cows typically experience immunosuppression with dysregulated neutrophil function (e.g., compromised phagocytosis) during the transition period (3 wk before to 3 wk after parturition), which is causally associated with increased risk of infections. Enhanced neutrophil immune competence has significant bearing with the wellbeing of transition dairy cattle. In the current study, we investigated the effect of OmniGen-AF (OG; Phibro Animal Health, Quincy, IL) and its dose in modulation of neutrophil function of transition cows. Forty-eight multiparous cows were stratified by parity, somatic cell count, and expected calving date and randomly assigned to 3 treatments: OG fed at 0 g/head per day (CON), 60 g/head per day (OG60; recommended dose), and 90 g/head per day (OG90; 1.5× recommended dose). The OG was added from dry off (61.8 ± 1.69 d before parturition) to 28 d in milk (DIM), and removed from all treatment groups at 29 to 35 DIM (the last week of the experimental period). Neutrophil phagocytic ability against Staphylococcus aureus and Escherichia coli was improved and tended to be improved, respectively, by OG from d 28 before parturition to 28 DIM. Cows in OG60 had higher neutrophil phagocytic ability against S. aureus and E. coli compared with CON cows from d 28 before parturition to 28 DIM. Neutrophil phagocytosis of S. aureus and E. coli was higher and tended to be higher for OG60 compared with CON on 35 DIM. The relative gene expression of CXCL8 and SELL were upregulated and tended to be upregulated by OG from 60 d before parturition to 28 DIM; this was due to cows in OG60 having greater SELL and CXCL8 gene expression than CON. Expression of SELL in circulating white blood cells of OG60-treated cows was greater than OG90 and the relative expression of CXCL8 gene tended to be greater for OG60 compared with CON on 35 DIM. In conclusion, feeding OG at the recommended dose of 60 g/head per day from dry off was effective in maintaining peripheral blood neutrophil function in transition dairy cows, and it is not necessary to feed OG beyond the recommended dose., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Intramuscular fat (IMF) content has been generally recognized as a desirable trait in pork meat because of its positive effect on eating quality. An effective approach to enhance IMF content in pork is the generation of transgenic pigs. In this study, we used somatic cell nuclear transfer (SCNT) to generate cloned pigs exhibiting ectopic expression of phosphoenolpyruvate carboxykinase (PEPCK-C) driven by an α-skeletal-actin gene promoter, which was specifically expressed in skeletal muscle. Using qRT-PCR and Western blot analysis, we demonstrated that PEPCK-C was functionally expressed and had a significant effect on total fatty acid content in the skeletal muscle of the transgenic pigs, while the n-6/n-3 polyunsaturated fatty acid (PUFA) ratio showed no difference between transgenic and control pigs. Thus, genetically engineered PEPCK-C mus pigs may be an effective solution for the production of IMF-enriched pork.

The postnatal neurodevelopmental disorder Rett syndrome, caused by mutations in MECP2, produces a diverse array of symptoms, including loss of language, motor, and social skills and the development of hand stereotypies, anxiety, tremor, ataxia, respiratory dysrhythmias, and seizures. Surprisingly, despite the diversity of these features, we have found that deleting Mecp2 only from GABAergic inhibitory neurons in mice replicates most of this phenotype. Here we show that genetically restoring Mecp2 expression only in GABAergic neurons of male Mecp2 null mice enhanced inhibitory signaling, extended lifespan, and rescued ataxia, apraxia, and social abnormalities but did not rescue tremor or anxiety. Female Mecp2(+/-) mice showed a less dramatic but still substantial rescue. These findings highlight the critical regulatory role of GABAergic neurons in certain behaviors and suggest that modulating the excitatory/inhibitory balance through GABAergic neurons could prove a viable therapeutic option in Rett syndrome.

Fatty acid synthase (FASN), a lipogenic multi-enzyme complex, is reported to be overexpressed in various types of of tumor tissues and serves an important role in tumor development and progression. However, the expression of FASN and its possible role in gastric cancer (GC) remains to be defined. In the present study, FASN expression in a group sample of 167 GC tissues was detected by immunohistochemistry and its correlation with clinicopathological features was analyzed. By clinical analysis, it was identified that FASN overexpression was positively correlated with the overall survival [P=0.008; hazard ratio (HR), 4.412; 95% confidence interval (CI), 1.463‑13.305] and recurrence rate (P=0.014; HR, 1.705; 95% CI, 1.116‑2.606) in patients with GC. In addition, expression of the FASN protein in GC tissues was correlated with age (P=0.032), clinical stage (P<0.001), gastric wall invasion (P=0.014), lymph node metastasis (P<0.001) and distant metastasis (P<0.001), however not with gender (P>0.05). In addition, FASN was observed to be overexpressed in GC tissues at an mRNA and protein level, compared with the adjacent non-cancerous tissues (P<0.05). Taken together, it was suggested that FASN was closely associated with GC metastasis and survival, which further provided evidence that FASN may be a promising prognostic biomarker for patients with GC.

Cadmium (Cd) is a toxic metal which harms human health through food chains. The mechanisms underlying Cd accumulation in oilseed rape are still poorly understood. Here, we investigated the physiological and genetic processes involved in Cd uptake and transport of two oilseed rape cultivars (Brassica napus). L351 accumulates more Cd in shoots but less in roots than L338. A scanning ion-selective electrode technique (SIET) and uptake kinetics of Cd showed that roots were not responsible for the different Cd accumulation in shoots since L351 showed a lower Cd uptake ability. However, concentration-dependent and time-dependent dynamics of Cd transport by xylem showed L351 exhibited a superordinate capacity of Cd translocation to shoots. Additionally, the Cd concentrations of shoots and xylem sap showed a great correlation in both cultivars. Furthermore, gene expression levels related to Cd uptake by roots (IRT1) and Cd transport by xylem (HMA2 and HMA4) were consistent with the tendencies of Cd absorption and transport at the physiological level respectively. In other words, L351 had stronger gene expression for Cd transport but lower for Cd uptake. Overall, results revealed that the process of Cd translocation to shoots is a determinative factor for Cd accumulation in shoots, both at physiological and genetic levels., (Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.)

Background and Objective: It has been proven that nm23-H1 gene was a tumor metastatic suppressor gene. However, it’s molecular mechanism of suppressing metastasis remains unexplored. There is a closely relationship between the abnormality of stucturs and functions of nm23-H1 gene, and cancer invasion and metastasis. We have constructed the vector with nm23-H1-shRNA and the vector with nm23-H1cDNA resistant to the specific shRNA. So, we plan to construct shRNA-resistant eukaryotic expression vector of nm23-H1 gene by site-directed mutagenesis, rescue experiment was performed to verify the nm23-H1 gene expression, and to provide basement for studying the biochemical mechanisms of nm23-H1 gene., Methods: Site-directed mutagenesis of nm23-H1 gene was performed by overlap extension PCR method. Pure plasmid containing gene of nm23-H1 (shRNA-resistant) was prepared. The desired five mutations were constructed and cloned into the eukaryotic vector pcDNA3.1Hygro(+). The human lung adenocarcinoma cell A549/nm23-H1-shRNA (stable nm23-H1 gene silencing) was transfected with the five mutants, and the expression of the mutant proteins was determined by Western blot., Results: Five eukaryotic expression vectors (shRNA-resistant) of nm23-H1, nm23-H1S44A, nm23-H1(P96S), nm23-H1(H118F), nm23-H1(S120G), nm23-H1(P96S-S120G), were successfully constructed. The results of DNA sequencing confirmed that the base sequences of the genes were completely concordant with experiment design. The expression of nm23-H1 mutant proteins was verified by Western blot., Conclusion: Five eukaryotic expression vectors (shRNA-resistant) of nm23-H1 gene were successfully constructed, and the mutant proteins were verified. The site-directed mutagenesis technical of overlap extension PCR is a efficient, simple and economical method.

Background and Objective: On the condition that laboratory animals survive, we can detect the distribution of tumor cells by in vivo imaging that were labeled with firefly- luciferase (luc) gene. The purpose of this study is to establish a light-emitting cell line A549/nm23-H1-shRNA-luc which could express nm23-H1 shRNA and firefly-luciferase stably, and detect its bioluminescence in vitro and in vivo. It will provide the preparation for the next related experimental research in vivo., Methods: The optimal concentration of hygromycin B for screening A549/nm23-H1-shRNA cells was determined by concentration gradient method. We firstly transfected the plasmid (PGL4.50) with luc gene into A549/nm23-H1-shRNA cells and then screened the monoclonal cell line A549/nm23-H1-shRNA-luc with hyhromycin B. The positive monoclonal cell line was identified with an in vivo imaging system, thereafter the expression stability of luciferase was analyzed in the strongest light-emitting positive monoclonal cell line. The A549/nm23-H1-shRNA-luc cells were inoculated subcutaneously into right-hind groin of nude mice and then observed by the in vivo imaging system., Results: The optimal concentration of hygromycin B used in screening A549/nm23-H1-shRNA cells was 300 μg/mL. After screening, the A549/nm23-H1-shRNA-luc cells established can express luciferase stably in vitro, a great linear correlation existed between the amount of cells (x) and bioluminescence values (y), with an equation of y=3,699.9x+992,237, and the square of the correlation coefficient (R2) was 0.975,1. To evaluate the stability of bioluminescence in vivo, 10 nude mice were randomly divided into two groups that the same number of cells were implanted into. The variation of bioluminescence values detected in vivo between the two groups of the same cells was not statistically significant (P>0.05)., Conclusions: We have successfully established the cell line A549/nm23-H1-shRNA-luc which can express luciferase persistently and stably.

Bone morphogenetic protein 11 (BMP11) is a key regulatory protein in skeletal development. BMP11 propeptide has been shown to antagonize GDF11 activity in vitro. To explore the role of BMP11 propeptide in skeletal formation in vivo, we generated transgenic mice with skeleton-specific overexpression of BMP11 propeptide cDNA. The mice showed a transformation of the seventh cervical vertebra into a thoracic vertebra in our previous report. Presently, further characterizations of the transgenic mice indicated that ossification in calvatia was dramatically enhanced in transgenic fetuses at 16.5 dpc in comparison with their wild-type littermates. At 10 weeks of age, bone mineral content and bone mineral density were significantly (P<0.05) higher in transgenic mice than that in their wild-type littermates based on dual energy X-ray absorptiometry analysis. The relative trabecular bone volume measured by histological analysis was dramatically increased in transgenic mice compared with their wild-type littermates. The enhanced bone formations in the transgenic mice appear to result from increase osteoblast activities as the expressions of four osteoblast markers - α1 type 1 collagen, osteocalcin, alkaline phosphatase and phex were significantly higher in transgenic fetuses than that in their wild-type littermates. These results suggest that over-expression of BMP11 propeptide stimulates bone formation by increasing osteoblast cell functions., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objective: To study the influence of aconite root, a Chinese medicinal herb with hot property, on energy metabolism and gene expression spectrum, and to analyze the possible mechanism of it effect., Method: Thirty two SPF Wistar rats were randomly divided into aconite root group and control group. Decoction of aconite root and NS were intragastrically administrated with the concentration of 10 mL x kg(-1) respectively once a day for 20 days. Temperature, energy intake (EI), digestive energy (DE) and metabolic energy (ME) were measured. The activity of ATPase and succinate dehydrogenase (SDH) in liver was detected by colorimetry. The gene expression of liver was detected with Illumina's rat ref-12 gene array. The differential expression genes were selected, annotated and classified based on gene ontology (GO). Real-time quantitative reverse-transcriptase PCR (Q-RT-PCR) was used to test the accuracy of the array results., Result: Compared with the control group, the toe temperature (TT) on the 10th and 20th day after the administration,the EI/BM( body mass), DE/BM, ME/BM and the activity of Na+ - K+ - ATPase, Ca2+ - Mg2+ - ATPase and SDH of liver in the aconite root group increased significantly (P<0.05). There were 592 differential expression genes in aconite root group compared with the control group. Based on Go analysis, the most significant genes was related to metabolic process (lgP = - 15.5897)., Conclusion: Aconite root could improve the energy metabolism in rats, by influencing the metabolic process of sugar, lipid and amino acid, which may be the main molecular mechanism of warming yang and dispelling cold for the treatment of the cold syndrome according to Chinese medicine theory.

Toll-like receptor 4 (TLR4), which recognizes lipopolysaccharide from Gram-negative bacteria, plays a major role in resistance of mice and humans to Salmonella infection. In chickens, Salmonella may establish a carrier state whereby bacteria are able to persist in the host organism without triggering clinical signs. Based on cellular morphological parameters, we developed a method, without using antibodies, to separate three cecal cell subpopulations: lymphocytes, enterocytes, and a population encompassing multiple cell types. We analyzed the mRNA expression of TLR4, interleukin-1β (IL-1β), IL-8, IL-12, and lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF) in cecal subpopulations of chicks from inbred lines resistant or susceptible to the carrier state infected with Salmonella enterica serovar Enteritidis. The results showed that resistance to the carrier state in chicks is associated with a larger percentage of lymphocytes and with higher levels of expression of TLR4 and IL-8 at homeostasis in the three cell subpopulations, as well as with a higher level of expression of LITAF in lymphocytes during the carrier state. In contrast to the early phase of infection, the carrier state is characterized by no major cell recruitment differences between infected and noninfected animals and no significant modification in terms of TLR4, IL-1β, IL-8, IL-12, and LITAF expression in all cell subpopulations measured. However, TLR4 expression increased in the lymphocytes of chicks from the susceptible line, reaching the same level as that in infected chicks from the resistant line. These observations suggest that the carrier state is characterized by a lack of immune activation and highlight the interest of working at the level of the cell population rather than that of the organ.

Aims: The main aims of this study were to clone and express flagellin flaA gene from Vibrio alginolyticus strain HY9901, also to prepare mouse anti-FlaA polyclonal antibody for future pathogen or vaccine study., Methods and Results: The full-length flaA gene was amplified by PCR with designed primers. The open reading frame of flaA gene contains 1131 bp, and its putative protein consists of 376 amino acid residues. Alignment analysis indicated that the FlaA protein was highly conserved. SDS-PAGE indicated that the FlaA protein was successfully expressed in Escherichia coli BL21 (DE3). Then, the recombinant FlaA protein was purified by affinity chromatography, and the mouse anti-FlaA serum was produced. The expression of flaA gene was verified by various immunological methods, including western blotting, enzyme-linked immunosorbent assay (ELISA) and immunogold electron microscopy (IEM)., Conclusions: Flagellin flaA gene was cloned and identified from V. alginolyticus HY9901, the recombinant FlaA protein was expressed and purified, and high-titre FlaA protein-specific antibody was produced. Western blot analysis revealed that the prepared antiserum not only specifically react to FlaA fusion protein, but also to natural FlaA protein of V. alginolyticus. The expressed FlaA protein was demonstrated, for the first time, as the component of flagella from V. alginolyticus by IEM., Significance and Impact of the Study: This study may offer important insights into the pathogenesis of V. alginolyticus, provide a base for further studies on the diagnosis and evaluation that whether the FlaA protein could be used as an effective vaccine candidate against infection by V. alginolyticus and other Vibrio species. Additionally, the purified FlaA protein and polyclonal antibody can be used for further functional and structural studies.

The present study investigated the effect of curcumin, a phenolic compound with yellow color from Curcuma longa L., on the expression of the apoptosis-related genes [BAX (Bcl-2 associated protein X), PKB, p53, MDM2 (mouse double minute 2), caspase 9, c-Ski, smad1 and smad4] in hamster opisthorchiasis. On Opisthorchis viverrini infection treated with dietary curcumin apoptosis-related gene expression profiles were similar to O. viverrini-infected group, but the expression levels seemed lower. Light microscopic observation revealed that aggregation of inflammatory cells surrounding the hepatic bile ducts in the groups infected with O. viverrini and treated with dietary curcumin was lower than in infected group. The intensity of the response is correlated with expression of the genes studied. The results suggest that curcumin reduces pathogenesis in hamster-opisthorchiasis by controlling apoptosis-related gene expression.

Pygopus, a very important component of the Wnt signaling transcriptional complex, has multiple functions in both Wnt-dependent and -independent pathways. Human Pygopus2 (Pygo2) is expressed in many cancers and plays an important role in tumor growth. In the present study, we generated human carcinoma (HeLa) cell lines stably expressing Pygo2, which counteracts vinblastine- induced apoptosis. The anti-apoptotic function was determined by DNA fragmentation, sub-G1 appearance, loss of mitochondrial membrane potential (Deltapsim) and the activation of caspase-9 and caspase-3. In addition, we found that Pygo2 effectively blocks vinblastineinduced c-Jun and AP-1 activation, maintains the anti-apoptotic protein Bcl-2 in an unphosphorylated state, and thus can render cells resistant to apoptosis. However, Pygo2 does not alter the vinblastine-induced cell cycle changes. Here, we describe an anti-apoptotic activity exerted by Pygo2 through blocking activation of the JNK/AP-1 signaling pathway induced by vinblastine.

Many ligands that affect nervous system development are members of gene families that function together to coordinate the assembly of complex neural circuits. cpg15/neuritin encodes an extracellular ligand that promotes neurite growth, neuronal survival, and synaptic maturation. Here we identify cpg15-2 as the only paralogue of cpg15 in the mouse and human genome. Both genes are expressed predominantly in the nervous system, where their expression is regulated by activity. cpg15-2 expression increases by more than twofold in response to kainate-induced seizures and nearly fourfold in the visual cortex in response to 24 hours of light exposure following dark adaptation. cpg15 and cpg15-2 diverge in their spatial and temporal expression profiles. cpg15-2 mRNA is most abundant in the retina and the olfactory bulb, as opposed to the cerebral cortex and the hippocampus for cpg15. In the retina, they differ in their cell-type specificity. cpg15 is expressed in retinal ganglion cells, whereas cpg15-2 is predominantly in bipolar cells. Developmentally, onset of cpg15-2 expression is delayed compared with cpg15 expression. CPG15-2 is glycosylphosphatidylinositol (GPI) anchored to the cell membrane and, like CPG15, can be released in a soluble-secreted form, but with lower efficiency. CPG15 and CPG15-2 were found to form homodimers and heterodimers with each other. In hippocampal explants and dissociated cultures, CPG15 and CPG15-2 promote neurite growth and neuronal survival with similar efficacy. Our findings suggest that CPG15 and CPG15-2 perform similar cellular functions but may play distinct roles in vivo through their cell-type- and tissue-specific transcriptional regulation., ((c) 2008 Wiley-Liss, Inc.)

To investigate the relationship between the thickness of tongue fur, apoptosis of the tongue fur epithelial cells and expressions of apoptosis-related genes in diseases of the digestive system, apoptosis-related genes TGF-beta3, fas mRNA and protein products were detected with terminal deoxynucleotidyl transferase-mediated deoxyurine triphosphate (d-UTP) nick-end labeling (TUNEL) technique, in situ hybridization, immunohistochemical methods, and image analysis technique, respectively. Results indicated that compared with the normal tongue fur, over-expression of fas gene was found in the peeling fur with an increase in cell apoptosis, while a low-expression of TGF-beta3 in the thick fur with a decrease in cell apoptosis. The changes in expression levels of fas and TGF-beta3 genes, apoptosis-promoting genes in the tongue fur epithelial cells, had a similar tendency of cell apoptosis level. It is concluded that the changes in expression levels of fas and TGF-beta3 are possibly important reasons influencing apoptosis of epithelial cells of tongue fur and leading to changes in thickness of the tongue fur.

Expression of the human DNA topoisomerase IIalpha (topo IIalpha) gene is positively regulated by the binding of the nuclear factor Y (NF-Y) transcription factor to four of five inverted CCAAT boxes (ICBs) located in its promoter. We have demonstrated previously that expression of the p53 tumor suppressor inhibits human topo IIalpha promoter activity in murine (10)1 cells. In this report, we demonstrate that the inhibition of topo IIalpha gene expression by wild-type p53 correlates with the decreased binding of the transcription factor NF-Y to the first four ICBs of the topo IIalpha promoter. The expression of mutant p53 does not affect the binding of NF-Y. In NIH3T3 cells, we show that topo II-targeted drugs inhibit the binding of NF-Y to ICB sites in the topo IIalpha promoter. This effect is seen not only with drugs that result in DNA strand breaks but also with drugs that inhibit the catalytic activity of topo II, and even with the mitotic spindle inhibitor, vinblastine. Further experiments with p53-null (10)1 cells treated with these same drugs also demonstrate decreased NF-Y binding to the topo IIalpha ICBs. The data presented points to the existence of both p53-dependent and -independent mechanisms for regulating NF-Y binding to ICBs in the topo IIalpha promoter and thus the modulation of topo IIalpha gene expression.

Objective: To investigate the killing effect of herpes simplex virus thymidine kinase gene expression actuated by the promoter of the vascular endothelial growth factor gene on human hepatocellular tumor cells under hypoxic condition., Methods: Recombinant adenoviral vectors, AdVEGF-tk and AdVEGF-GFP, were constructed with HSV-tk or GFP under the control of VEGF promoter through AdEasy system. Then GFP expression in hepatoma cell line HepG2 and normal liver cell line L02 transfected with AdVEGF-GFP were observed under fluorescence microscope, and the sensitivity to GCV of the AdVEGF-tk-transfected cells under normoxia or hypoxia condition were monitored by MTT method., Results: GFP expression actuated by VEGF promoter was detected in sporadic L02 cells, but in almost all HepG2 cells after transfected with AdVEGF-GFP. With GCV at 10 micro g/ml and MOI at 100, L02 cells were insensitive to GCV under oxic condition, but more than 70% L02 cells were killed under hypoxic condition. Moreover, HepG2 cells infected with AdVEGF-tk showed the increased GCV sensitivity under hypoxia (over 80% killed) as compared with normoxia (over 60% killed) conditions., Conclusion: Hypoxia enhances the GCV sensitivity of human hepatocellular tumor cells infected with recombinant AdVEGF-tk under the control of VEGF promoter.

Background: The Rel/NF-kappaB transcription factor pathway, regulated by IkappaB proteins, is considered central to immune responses, although there are surprisingly few in vivo data concerning alloresponses., Methods: We undertook analysis of NF-kappaB and IkappaB mRNA intracardiac allograft expression, and NF-kappaB nuclear translocation, during acute rejection versus CD154 monoclonal antibody (mAb)-induced tolerance induction in fully MHC-disparate mice., Results: Intragraft expression of all nine NF-kappaB and IkappaB genes increased during development of rejection, and nuclear translocation of p50, p52, and p65 was detected. CD154 mAb therapy decreased mRNA levels of all nine NF-kappaB and IkappaB genes, and impaired nuclear translocation of p50, p52, and p65 NF-kappaB proteins. However, prolonged survival could not be induced by CD154 mAb in p50- or p52-deficient allograft recipients, indicating an absolute requirement for expression of these genes in CD154 mAb-induced tolerance., Conclusions: We conclude that, whereas blanket approaches to NF-kappaB suppression are unlikely to be effective strategies for tolerance induction, a better understanding of the roles of individual NF-kappaB and IkappaB genes may allow development of more precise and effective therapies.

Muscle tissue is the major site for insulin-stimulated glucose uptake in vivo, due primarily to the recruitment of the insulin-sensitive glucose transporter (GLUT4) to the plasma membrane. Surprisingly, virtually all cultured muscle cells express little or no GLUT4. We show here that adenovirus-mediated expression of the transcriptional coactivator PGC-1, which is expressed in muscle in vivo but is also deficient in cultured muscle cells, causes the total restoration of GLUT4 mRNA levels to those observed in vivo. This increased GLUT4 expression correlates with a 3-fold increase in glucose transport, although much of this protein is transported to the plasma membrane even in the absence of insulin. PGC-1 mediates this increased GLUT4 expression, in large part, by binding to and coactivating the muscle-selective transcription factor MEF2C. These data indicate that PGC-1 is a coactivator of MEF2C and can control the level of endogenous GLUT4 gene expression in muscle.

Objective: To investigate whether apoptotic injury is induced by Fas mRNA expression mediated by H(2)O(2) and concerned with intracellular Ca(2+) unbalance at a single cell level., Methods: Fas mRNA expression in human hepatocyte (LO(2) cell) was analyzed by whole-cell patch-clamp combined with single cell reverse transcription polymerase chain reaction (RT-PCR). Cytosolic Ca(2+) concentration was measured with another whole-cell patch-clamp combined with single-cell microfluorescence [Ca(2+)] i measurement at the same time. The index of early phase apoptotic cells (Annexin-V(+) cells) measured with flow cytometry (FCM) and the hepatocytic morphological changes were observed by scanning electron microscopy., Results: Fas mRNA of LO(2) cells cultured with H(2)O(2) (10 micromol/L) was expressed at 2 h. The single cell [Ca(2+)] i analyzed by another whole-cell patchclamp increased to 1 115 nmol/L +/- 227 nmol/L (P < 0.01, t = 13.37); at the same time, an elevated Ca(2+) wave was recorded. The index of Annexin-V(+) cells was as high as 16.18 +/- 0.65 (P < 0.01, t = 48.41), and the membrane surface protuberances were observed., Conclusions: The activation of Fas mRNA, as a major apoptotic factor mediated by H(2)O(2) in LO(2) cells, is probably associated with the imbalance of intracellular Ca(2+).

Soil acidification is common in some Chinese tea plantations, which seriously affected growth of tea trees. Hence, it is essential to explore soil remediation in acidified tea plantations for sustainable development of the tea industry. We sought to determine how different fertilizers affect acidified soil and their N transformation in tea plantations.Different fertilizers were used on acidified tea plantation soils for 4 years (2017–2021), and changes in soil pH, indices related to soil N transformation and tea yield were analysed to construct interaction networks of these indices and find which had the largest influence on fertilization.Long‐term use of sheep manure reduced soil acidification, increased soil pH, enhanced the number and intensity of N‐fixing and ammonifying bacteria, urease, protease, asparaginase and N‐acetamide glucose ribosidase activity and nifH gene expression. This treatment reduced the number and intensity of soil nitrifying and denitrifying bacteria, nitrate reductase and nitrite reductase activity, while the expression of amoA‐AOA, nirK, nirS, narG and nosZ in turn increased ammonium N content of the soil, reduced nitrate N content, and enhanced tea yield. Topsis index weight analysis showed that ammonium N content in the soil had the largest impact among fertilization effects.Long‐term use of sheep manure was beneficial in restoring the balance of the micro‐ecosystem in acidified soil. This study provides an important practical basis for soil remediation and fertilizer management in acidified tea plantation soils. [ABSTRACT FROM AUTHOR]

Immune checkpoint blockade (ICB) targeting PD-1/PD-L1 has been used for the treatment of head and neck squamous cell carcinoma (HNSCC). However, the overall response rate to ICB therapy for HNSCC remains less than 20%. It has recently been reported that the appearance of tertiary lymphoid structures (TLSs) in tumor tissue is correlated with better prognosis and response to ICB treatment. Here, we demonstrated an immune classification for the tumor microenvironment (TME) of HNSCC by analyzing The Cancer Genome Atlas (TCGA)–HNSCC data set and found that immunotype D with TLS enrichment had a better prognosis and response to ICB treatment. Furthermore, we observed that TLSs were present in a part of tumor samples of human papillomavirus (HPV) infection negative HNSCC (HPV− HNSCC) and were associated with the densities of dendritic cell (DC)–LAMP+ DCs, CD4+ T cells, CD8+ T cells, and progenitor T cells in TME. We established an HPV− HNSCC mouse model with TLS-enriched TME by overexpressing LIGHT in a mouse HNSCC cell line. We found that the induction of TLS formation enhanced the response to PD-1 blockade treatment in the HPV− HNSCC mouse model, accompanied by increases in DCs and progenitor exhausted CD8+ T cells in the TME. Elimination of CD20+ B cells attenuated the therapeutic effect of PD-1 pathway blockade in TLS+ HPV− HNSCC mouse models. These results indicate that TLSs contribute to the favorable prognosis and antitumor immunity of HPV− HNSCC. Inducing TLS formation in HPV− HNSCC tumors is a potential therapeutic method for improving the ICB response rate in patients with HPV− HNSCC. [ABSTRACT FROM AUTHOR]

Populus euphratica belongs to Salicaceae family and grows in extreme desert environments. At present, the identification of the AP2/ERF gene family of transcription factors in Salicaceae is rare, and the role of the AP2/ERF gene family in P. euphratica under salt stress and exogenous hormones has not been reported. In this study, 197, 210, 231, 192, and 147 AP2/ERF genes were identified in P. euphratica, Populus trichocarpa, Populus deltoides, Salix sinopurpurea, and Arabidopsis thaliana, respectively. The 197 AP2/ERF gene family members of P. euphratica were divided into five subfamilies, namely, AP2 (35), RAV (5), ERF (96), DREB (65), and Soloist (1), by sequence alignment and phylogenetic analysis. In addition, these genes were scattered across 19 chromosomes. The detection of 10 motifs in the P. euphratica AP2/ERF gene family revealed that motif-8 and motif-9 only appeared in the ERF subfamily and DREB subfamily, respectively. Transcriptome data showed that PeAP2/ERF genes had different expression patterns under salt stress, abscisic acid (ABA) and gibberellic acid (GA3) treatments, suggesting that the genes PeERF002, PeERF037, PeERF082, PeERF090, and PeAP2-14 may play important roles under salt stress and exogenous hormone treatments. This study provides a reference for the functional study of the PeAP2/ERF gene, and it also lays a foundation for the breeding strategy to improve the salt tolerance of P. euphratica. [ABSTRACT FROM AUTHOR]

To evaluate the impacts of guanidinoacetic acid (GAA) and coated folic acid (CFA) on growth performance, nutrient digestion and hepatic gene expression, fifty-two Angus bulls were assigned to four groups in a 2 × 2 factor experimental design. The CFA of 0 or 6 mg/kg dietary DM folic acid was supplemented in diets with GAA of 0 (GAA−) or 0·6 g/kg DM (GAA+), respectively. Average daily gain (ADG), feed efficiency and hepatic creatine concentration increased with GAA or CFA addition, and the increased magnitude of these parameters was greater for addition of CFA in GAA− diets than in GAA+ diets. Blood creatine concentration increased with GAA or CFA addition, and greater increase was observed when CFA was supplemented in GAA+ diets than in GAA− diets. DM intake was unchanged, but rumen total SCFA concentration and digestibilities of DM, crude protein, neutral-detergent fibre and acid-detergent fibre increased with the addition of GAA or CFA. Acetate:propionate ratio was unaffected by GAA, but increased for CFA addition. Increase in blood concentrations of albumin, total protein and insulin-like growth factor-1 (IGF-1) was observed for GAA or CFA addition. Blood folate concentration was decreased by GAA, but increased with CFA addition. Hepatic expressions of IGF-1, phosphoinositide 3-kinase, protein kinase B, mammalian target of rapamycin and ribosomal protein S6 kinase increased with GAA or CFA addition. Results indicated that the combined supplementation of GAA and CFA could not cause ADG increase more when compared with GAA or CFA addition alone. [ABSTRACT FROM AUTHOR]

Aims: Relatively, few anti‐biofilm polysaccharides against Pseudomonas aeruginosa were done to investigate the underlying molecular mechanism. Exopolysaccharide EPS273 can clearly reduce biofilm formation and infection of P. aeruginosa. This study aims to investigate its anti‐biofilm and anti‐infection mechanism on transcriptional level. Methods and Results: Herein, we used an RNA‐Seq transcriptomic approach to investigate the underlying anti‐biofilm and anti‐infection mechanism of EPS273. The expression levels of a large number of genes were changed after P. aeruginosa PAO1 was treated with EPS273. Especially, the genes related to biofilm formation, such as gene involved in production of extracellular matrix and virulence factor, genes involved in flagella and cell motility and genes involved in iron acquisition. Notably, the expression levels of genes involved in regulatory and signal transduction were markedly downregulated, such as two‐component system PhoP‐PhoQ and quorum sensing (QS) system LasI/LasR and RhlI/RhlR. Furthermore, when genes phoP and phoQ were disrupted, respectively, the reduction of biofilm formation and cell motility in mutant △phoP or △phoQ was also detected. Conclusion: EPS273 may exert its anti‐biofilm and anti‐infection function by downregulating gene expression of two‐component system PhoP‐PhoQ and QS systems LasI/LasR and RhlI/RhlR of P. aeruginosa, which further regulated expression of genes involved in biofilm formation. Significance and Impact of the Study: Our data will expand understanding of anti‐biofilm mechanisms of polysaccharides on transcriptomic level. [ABSTRACT FROM AUTHOR]

In this study, melatonin (MEL)‐mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti‐virus infection activity to 37.4% in virus‐infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT‐PCR and virus titres measured by dot‐ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT‐PCR, in virus‐infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H2O2) in the virus‐infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL‐induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT‐PCR and virus titres analysed by dot‐ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL‐mediated plant resistance to TMV is proposed. MEL‐mediated plant resistance to viruses provides a new avenue to control plant viral diseases. [ABSTRACT FROM AUTHOR]

Species-specific cusp patterns result from the iterative formation of enamel knots, the epithelial signaling centers, at the future cusp positions. The expressions of fibroblast growth factors (FGFs), especially Fgf4, in the secondary enamel knots in the areas of the future cusp tips are generally used to manifest the appearance of species-specific tooth shapes. However, the mechanism underlying the predictive role of FGFs in species-specific cusp patterns remains obscure. Here, we demonstrated that gerbils, which have a lophodont pattern, exhibit a striped expression pattern of Fgf4, whereas mice, which have a bunodont pattern, have a spotted expression pattern, and these observations verify the predictive role of Fgf4 in species-specific cusp patterns. By manipulating FGFs' signaling in the inner dental epithelium of gerbils, we provide evidence for the intracellular participation of FGF signaling, specifically FGF4 and FGF20, in Rac1- and RhoA-regulated cellular geometry remolding during the determination of different cusp patterns. Our study presents a novel explanation of how different FGF expression patterns produce different cusp patterns and implies that a conserved intracellular FGF-GTPase signaling module might represent an underlying developmental basis for evolutionary changes in cusp patterns. [ABSTRACT FROM AUTHOR]

BACKGROUND: Analysis using publicly available algorithms has found that high mobility group AT-hook 2 (HMGA2), a key transcriptional regulatory factor, is a potential target of microRNA-204 (miR-204). Some studies have shown that both miR-204 and HMGA2 are associated with cancer development. OBJECTIVE: We examined the possible relationship between miR-204 and HMGA2 in the development of thyroid cancer. METHODS: We assessed miR-204 expression in thyroid cancer specimens and cell lines using real-time polymerase chain reaction (PCR). Luciferase reporter assay was used to confirm the target associations. The effect of miR-204 on cell proliferation was confirmed with tetrazolium and colony formation assays. Gene and protein expression were examined using real-time PCR and western blotting, respectively. RESULTS: MiR-204 was downregulated in the thyroid cancer specimens and cell lines, and targeted the 3' untranslated region of HMGA2 directly. MiR-204 overexpression decreased cyclin D1 and Ki67 expression and increased P21 expression, which subsequently inhibited thyroid cancer cell proliferation. CONCLUSIONS: Our findings suggest that miR-204 plays a protective role by inhibiting thyroid cancer cell proliferation, and may identify new targets for anti-cancer treatment. [ABSTRACT FROM AUTHOR]

Post-mortem muscle energy metabolism plays an important role in pork quality. To analyse the differences of meat quality and energy metabolism, three commercial pig crossbreeds frequently used in China were studied, they were DT (Duroc x Taihu; n = 16), PIC (five-way crossbreed from Pig Improvement Co., UK; n = 29) and DLY (Duroc x (Landrace x Yorkshire); n = 19) pigs. The results showed that DT pigs had a higher post-mortem pH 45 min and pH24 h, lower shear force and drip loss, higher muscle free-glucose and glycogen contents, and lower lactic acid content than did PIC and DLY pigs. Post-mortem muscle free-glucose content of these three pig crossbreeds changed little, from 45 min to 96 h post-mortem. The expression levels of PRKAG3 (encoding a regulatory subunit of the AMP-activated protein kinase) and GYS1 (encoding muscle glycogen synthase) genes of DT pigs were significantly lower than those of PIC and DLY pigs. DT pigs had a higher expression level of glycogenin-1-like (encoding glycogenin) gene than did PIC and DLY pigs. In conclusion, DT pigs had better meat quality than did the other two pig crossbreeds. We deduced that the post-mortem muscle energy status and metabolism of DT pigs might be an important reason for their good meat quality, and future research should focus on the molecular and physiological mechanism of post-mortem muscle energy metabolism to find ways to improve meat quality. Pork makes a major contribution to worldwide meat consumption, and ways of improving meat quality are imperative. Variation in the rate of post-mortem pH decline is an important reason for variation in meat quality, but the mechanisms involved are not clear. In our research, we found that post-mortem energy status and metabolism might be important factors in the conversion of muscle to meat and might be important determinants of ultimate meat quality. [ABSTRACT FROM AUTHOR]

The mu-opioid receptor ( MOR), a membrane-bound G protein-coupled receptor, is the main target for opioids in the nervous system. MOR1 has been found in several types of cancer cells and reported to be involved in tumor progression and metastasis. However, the expression and clinical significance of MOR1 in esophageal squamous cell carcinoma ( ESCC) remain unclear. In our study, the expression of MOR1 was confirmed in ESCC cell lines ( KYSE180, KYSE150, and EC109) by Western blot. MOR1 was also detected on tissue microarrays of ESCC samples in 239 cases using immunohistochemical staining. We found that MOR1 was mainly located in the cytoplasm and occasionally occurred in the membrane or nucleus of ESCC cells. Moreover, results indicated that MOR1 expression in the cytoplasm was associated with lymph node metastasis ( R = 0.164, P = 0.008, Kendall's tau-b-test). No more associations were found between MOR1 expression status and other clinical parameters. However, no statistical significant differences were found between MOR1 expression in the cytoplasm, nucleus/membrane, and the overall survival of ESCC patients ( P = 0.848; P = 0.167; P = 0.428, respectively, log-rank test). Our results suggest that the cytoplasmic MOR1 may be a high-risk factor for lymph node metastasis of ESCC patients. We also hypothesize that MOR1 agonists used in ESCC patients should be prudent, and opioid receptor antagonists may be novel therapeutic drugs for ESCC patients. [ABSTRACT FROM AUTHOR]

Aberrant expression of microRNAs (miRNAs) has been implicated in cancer initiation and progression. However, little is known about the potential role of miRNAs in glioma tumorigenesis. In this study, we found that miRNA-106b-5p was significantly upregulated in glioma tumor samples and cell lines compared with normal brain tissues, and its expression level correlated with the pathological grading. Overexpression of miR-106b-5p in glioma tumor cells significantly promoted cell proliferation, although inhibited cell apoptosis in vitro and in vivo. In contrast, knockdown of miR-106b-5p significantly inhibited cell proliferation, although enhanced cell apoptosis. Mechanistic study revealed that two target genes, retinoblastoma-like 1 (RBL1) and RBL2, were involved in miR-106b-5p's regulation of cell proliferation and one target gene, caspase-8 (CASP8), mediated miR-106b-5p's regulation of apoptosis. We also investigated the function of the three targets in glioma tumorigenesis by RNA interference manipulation and demonstrated that knockdown of these target genes led to cell proliferation enhancement or cell apoptosis inhibition in vitro. More interestingly, the expression levels of these targets were significantly downregulated in glioma samples and knockdown of these targets in glioma cells inhibited the xenograft tumor formation in vivo. Moreover, we verified the regulation function of miR-106b-5p and its targets on cell proliferation and apoptosis of the primary cultured astrocytes isolated from glioma tumor samples and healthy controls. Collectively, our findings show the critical roles of miR-106b-5p and its targets, RBL1, RBL2 and CASP8, in glioma tumorigenesis and provide potential candidates for malignant glioma therapy. [ABSTRACT FROM AUTHOR]

Angiostrongylus cantonensis ( A. cantonensis) infection is the major cause of eosinophilic meningitis (EM). Severe cases or infant and child cases have poor prognosis. MicroRNAs (miRNAs) play important roles in inflammation; however, little is known about the roles in brain inflammation caused by A. cantonensis. In this study, Illumina deep sequencing and bioinformatics were used to determine the abundance and differential expression of miRNAs in the brain tissues of a mouse model. A total of 648 conserved miRNAs were identified, 157 of which were significantly differentially expressed between infected mice and normal mice. The five most fold-changed miRNAs were miR-511-5p, miR-511-3p, miR-223-3p, miR-155-5p and miR-206-3p. These expressions of miR-511, miR-223, miR-155, miR-206, miR-142 and miR-21a were validated by quantitative reverse transcription polymerase chain reaction (RT-PCR). The analysis of these miRNAs showed that miR-511-3p was more abundant than the miR-511-5p strand, and increased to a peak in 21 days after A. cantonensis infection, miR-223 might be a potential indicator of disease severity and the upregulation of miR-155-5p after stimulation with the somatic antigen of phase IV A. cantonensis implied its involvement in the central nervous system (CNS) inflammation induced by A. cantonensis infection. These observations suggest that miRNAs may play important roles in the regulation of EM caused by A. cantonensis infection. [ABSTRACT FROM AUTHOR]

Background:HOX transcript antisense RNA (HOTAIR), which is expressed from the homebox C gene (HOXC) locus, is capable of reprogramming chromatin organisation and promoting cancer cell metastasis and can simultaneously bind the polycomb repressive complex 2, which enhances H3K27 trimethylation, and the LSD1-CoREST-REST complex, which is critical for H3K4 demethylation. Clinically, the overexpression of HOTAIR is a powerful predictor of the tumour progression and overall survival in patients with diverse cancers. The relationship between HOTAIR and oesophageal squamous cell carcinoma (ESCC), however, remains unclear. We investigated the role of HOTAIR in the pathogenesis of ESCC.Methods:We used quantitative real-time PCR to determine the level of HOTAIR in ESCC cell lines and 100 ESCC samples from patients; 56 adjacent non-neoplastic tissues were used as controls. We measured the effect of HOTAIR knockdown and overexpression in ESCC cell lines using colony formation assays, anchorage-independent growth assays, the CCK-8 assay, transwell migration and invasion assays, and Annexin V-binding assays. We analysed the growth of ESCC xenograft tumours in nude mice. Changes in the gene expression and methylation levels in ESCC cell lines were analysed using gene expression microarrays and the Infinium HumanMethylation450K BeadChip assay, respectively.Results:The levels of HOTAIR were increased in ESCC cell lines and patient samples compared with the controls; the expression levels correlated with the disease stage and survival time. The knockdown of HOTAIR in the KYSE510 and KYSE180 ESCC cell lines using small hairpin RNAs (shRNAs) reduced the ability of the cells to form foci, migrate, and invade the extracellular matrix in culture, altered cell cycle progression, and increased the sensitivity of the cells to apoptosis. The HOTAIR knockdown reduced cancer cell metastasis in vivo, and the tumours formed by HOTAIR-silenced ESCC cells were smaller, both in size and weight, than the tumours and metastases formed by the shRNA vector control cells in a mouse xenograft model. The results of the gene microarray study showed that HOTAIR reprogrammed the gene expression profile of ESCC cells, and the gene ontology analysis revealed an enrichment in genes that are important for tumorigenesis, such as genes involved in cell migration and the regulation of the cell cycle. Comparing the gene expression profiles and DNA methylation analysis between the KYSE180 and KYSE180_HOTAIR cells revealed that only a small proportion of the methylation changes were correlated with gene expression changes.Conclusion:HOX transcript antisense RNA is upregulated in ESCC cell lines and patient samples, and promotes ESCC cell proliferation and tumour metastasis in mice. The knockdown of HOTAIR resulted in significant changes in gene expression, and data analysis suggested that HOTAIR-mediated gene regulation has a critical role in ESCC progression and is a novel epigenetic molecular target for treating ESCC patients. [ABSTRACT FROM AUTHOR]

Abstract: Recent molecular studies have revealed that many genes are mobilized during nurse cell formation, including those involved in activation and proliferation of satellite cells, dedifferentiation, cell cycle re-entry and arrest, apoptosis and transformation. This article reviews the kinetics of gene expression from a cellular biology point-of-view in an effort to dissect the complex events that lead to unusual pathological changes after a Trichinella infection. [Copyright &y& Elsevier]

Calcium is an important element in water bodies that plays a pivotal role in numerous biological processes; however, its effects on bloom-forming cyanobacteria have been studied only rarely. In the present study, the effects of calcium onMicrocystis aeruginosa, a major bloom-forming cyanobacterium, were investigated. Significant decreases in growth, chlorophyllaand maximum electron transport rate were shown when calcium was absent, while similar reductions in these parameters were also found at high concentrations of calcium (240 mg/l). The expression ofpsbA,grpEandrecAwas up-regulated significantly for 0 and 240 mg/l calcium treatments. The activity of catalase (CAT) also increased remarkably under these two treatments, suggesting thatMicrocystissuffered from stress under both calcium absence and under high calcium concentrations. Additionally, the polysaccharide synthesis gene,epsL, was up-regulated at high concentrations of calcium. These data suggest that calcium plays an important role in the growth ofMicrocystis, and that enhancement in polysaccharide synthesis in response to increases in calcium concentration might be responsible for bloom formation. [ABSTRACT FROM PUBLISHER]

We previously reported regulated expression of erythropoietin (EPO) over 4 weeks in the peripheral nerve in vivo, using a herpes simplex virus (HSV)-based vector containing a Tet-on regulatable gene expression cassette. To create a vector that would be appropriate for the treatment of chronic neuropathy, we constructed a HSV vector with expression of EPO under the control of the Tet-on system in which the HSV latency-associated promoter 2 element was used to drive the expression of the Tet-on transactivator. EPO expression from the vector was tightly controlled by administration of doxycycline (DOX) in vitro. One month after inoculation of the vector to transduce dorsal root ganglion (DRG) in vivo, administration of DOX-containing chow-induced expression of EPO. Mice with streptozotocin-induced diabetes, inoculated with the vector, were protected against the development of neuropathy by continuous administration of DOX-containing chow over the course of 3 months. Identical results were achieved when DOX was administered every other week over 3 months of diabetes, but administration of DOX, 1 week out of 3, provided only partial protection against the development of neuropathy. Taken together, these results suggest such a vector is well suited for clinical trial for the treatment of chronic or subacutely developing neuropathy. [ABSTRACT FROM AUTHOR]

We evaluated the therapeutic effect of erythropoietin (EPO) delivered by direct injection of a nonreplicating herpes simplex virus (HSV)-based vector coding for EPO (vEPO) in a model of cervical hemicord contusion at C7. At 1 h after spinal cord injury (SCI), either vEPO or control vector carrying a reporter gene (vC) was injected into the cord above and below the lesion. Animals injected with vEPO showed a statistically significant improvement in the ipsilateral forelimb function, as measured by open-field evaluation of motor performance, forelimb reaching in the cylinder test and misplacement in grid walk. This correlated with preservation of gray matter in the area of the lesion. There was also mild but significant improvement of hindlimb motor function measured by Basso-Beattie-Bresnahan score and computerized gait analysis in vEPO compared with control vector-injected animals. Microtubule-associated protein tau, phosphorylated and nonphosphorylated neurofilament protein and the synaptic proteins synaptophysin and PSD-95 were all significantly increased in the spinal cord of vEPO-treated animals compared with control vector-injected animals. These data suggest that gene transfer of EPO after cervical SCI by minimizing the injury size and enhancing tissue sparing preserves large-caliber axons and promotes synaptogenesis. [ABSTRACT FROM AUTHOR]