Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome bac...Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome background. Transcriptomic analysis based on high-throughput RNA sequencing has been shown to be an efficient and powerful method for the discovery of relevant genes in non-model and genome reference-free organisms. Here, we constructed two c DNA libraries(control and Cd exposure groups) from the digestive gland of C. farreri and compared the transcriptomic data between them. A total of 227 673 transcripts were assembled into 105 071 unigenes, most of which shared high similarity with sequences in the NCBI non-redundant protein database. For functional classification, 24 493 unigenes were assigned to Gene Ontology terms. Additionally, Eu Karyotic Ortholog Groups and Kyoto Encyclopedia of Genes and Genomes analyses assigned 12 028 unigenes to 26 categories and 7 849 unigenes to five pathways, respectively. Comparative transcriptomics analysis identified 3 800 unigenes that were differentially expressed in the Cd-treated group compared with the control group. Among them, genes associated with heavy metal accumulation were screened, including metallothionein, divalent metal transporter, and metal tolerance protein. The functional genes and predicted pathways identified in our study will contribute to a better understanding of the metabolic and immune system in the digestive gland of C. farreri. In addition, the transcriptomic data will provide a comprehensive resource that may contribute to the understanding of molecular mechanisms that respond to marine pollutants in bivalves.展开更多
AIM: To investigate the expression and significance of caudal-related homeobox transcription factor (Cdx2) in gastric carcinoma (GC) and precancerous lesions. METHODS: The expression of Cdx2 in GC, precancer- ou...AIM: To investigate the expression and significance of caudal-related homeobox transcription factor (Cdx2) in gastric carcinoma (GC) and precancerous lesions. METHODS: The expression of Cdx2 in GC, precancer- ous lesions and normal gastric mucosa were detected using immunohistochemical method. Hematoxylin and eosin staining, alcian blue/periodic acid-schiff and high iron diamine/alcian blue staining were used to classify intestinal metaplasia (IM) and GC. RESULTS: Cdx2 was not detected in normal gas- tric mucosa. Cdx2 expression was detected in 87.1% (101/116) of IM, 50% (36/72) of dysplasia and 48.2% (41/85) of GC. The Cdx2-expressing cells in IM were more prevalent than in dysplasia and carcinoma (P 〈 0.05). There was no relationship between Cdx2 ex- pression and the classification of IM or the degree of dysplasia. Expression of Cdx2 was significantly higher in intestinal-type carcinoma than in diffuse and mixed- type carcinoma (P 〈 0.05). Positive expression of Cdx2was mainly found in moderately to well differentiated GC. There was a negative association between nuclear Cdx2 expression and lymph node metastasis and tumor, nodes, metastasis stage of GC (P 〈 0.05). The patients with Cdx2-positive expression showed a higher survival rate than those with Cdx2-negative expression (P = 0.038). Multivariate analysis revealed that the expres- sion of Cdx2 and lymph node metastasis were indepen- dent prognostic indicators of GC (P 〈 0.05).展开更多
OBJECTIVE To investigate the correlations between the expression of thyroid transcription factor-1 (TTF-1) and apoptosis and angiogenesis in lung carcinomas. METHODS A 829 microarray of the paraffin tissue chips was...OBJECTIVE To investigate the correlations between the expression of thyroid transcription factor-1 (TTF-1) and apoptosis and angiogenesis in lung carcinomas. METHODS A 829 microarray of the paraffin tissue chips was constructed, which contained 196 lung carcinomas, 10 normal lung tissues, and 1 muscular tissue. Terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) and immunohistochemical SP method were used to detect apoptosis and expression of TTF-1 and CD34 in different types of lung carcinomas. A Leica Q500 MC image analysis system was used to measure and calculate TTF-1 positive unit (PU), apoptotic index (AI) and microvessel density (MVD). RESULTS AI of lung small cell carcinoma and large cell carcinoma were smaller than those of lung adenocarcinoma and squamous cell carcinoma (P = 0.000). AI of lung carcinomas with lymph node metastases was smaller than that of those without (P = 0.039). AI of lung carcinomas in TNM stage I-IV was smaller than that in stage I (P = 0.008). The PU of the TTF-1 was negatively correlated with AI in small cell lung carcinoma (r = -0.752, P = 0.000). MVD of lung carcinomas without lymph node metastases was smaller than that of those with lymph node metastasis (P = 0.031). MVD of lung carcinomas in TNM stage I was smaller than that in stage I-IV (P = 0.040). The PU of TTF-1 was positively correlated with MVD in lung adenocarcinoma (r = 0.708, P = 0.000). CONCLUSION There is a negative correlation between TTF-1 PU and AI in small cell lung carcinoma. TTF-1 PU and AI may be correlated with each other. There is a positive correlation between TTF-1 PU and MVD in lung adenocarcinoma. TTF-1 may induce the development of lung adenocarcinoma by inducing tumor angiogenesis.展开更多
Calorie restriction(CR) promotes longevity among distinct organisms from yeast to mammals. Although CR-prolonged lifespan is believed to associate with enhanced respiratory activity, it is apparently controversial for...Calorie restriction(CR) promotes longevity among distinct organisms from yeast to mammals. Although CR-prolonged lifespan is believed to associate with enhanced respiratory activity, it is apparently controversial for accelerated energy consumption regardless of insufficient nutrient intake. In reconciling the contradiction of less food supply versus much metabolite dispense, we revealed a CR-based mode of dual-phase responses that encompass a phase of mitochondrial enhancement(ME) and a phase of post-mitochondrial enhancement(PME), which can be distinguished by the expression patterns and activity dynamics of mitochondrial signatures. ME is characterized by global antioxidative activation, and PME is denoted by systemic metabolic modulation. CR-mediated aging-delaying effects are replicated by artesunate, a semi-synthetic derivative of the antimalarial artemisinin that can alkylate heme-containing proteins, suggesting artesunate-heme conjugation functionally resembles nitric oxide-heme interaction. A correlation of artesunate-heme conjugation with cytochrome c oxidase activation has been established from adduct formation and activity alteration. Exogenous hydrogen peroxide also mimics CR to trigger antioxidant responses, affect signaling cascades, and alter respiratory rhythms, implying hydrogen peroxide is engaged in lifespan extension. Conclusively, artesunate mimics CR-triggered nitric oxide and hydrogen peroxide to induce antioxidative networks for scavenging reactive oxygen species and mitigating oxidative stress, thereby directing metabolic conversion from anabolism to catabolism, maintaining essential metabolic functionality, and extending life expectancy in yeast.展开更多
Objective: To differentiate rat adipose tissue-derived mesenchymal stem cells (ADSCs) into cells with a nucleus pulposus-like phenotype in vitro, so as to lay a foundation for the cell-based transplantation therapy...Objective: To differentiate rat adipose tissue-derived mesenchymal stem cells (ADSCs) into cells with a nucleus pulposus-like phenotype in vitro, so as to lay a foundation for the cell-based transplantation therapy of degenerated intervertebral discs. Methods: Rat ADSCs were isolated only from the subcutaneous inguinal region and purified by limited dilution. ADSCs of the third passages were analyzed by fluorescence activated cell sorter (FACS) to detect the cell surface markers (Sca-1, CD44, CD45, CDI lb). To induce ADSCs to- wards a nucleus pulposus-like phenotype, ADSCs were immobilized in 3-dimensional alginate hydrogels and cultured in an inducing medium containing transforming growth factor-beta1 (TGF- β1) under hypoxia (2% O2), while control groups under normoxia (21% O2) in alginate beads in medium with or without the presence of TGF-β 1. Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) was carried out to evaluate phenotypic and biosynthetic activities in the process of differentiation. Meanwhile, Alcian blue staining were used to detect the formation of sulfated glycosaminoglycans (GAGs) in the differentiated cells. Results: The purified ADSCs were fibroblast-like and proliferated rapidly in vitro. The flow cytometry showed that ADSCs were positive for Sca-1 and CD44, negative for CD45 and CD11b. The results of RT-PCR manifested that the gene expressions of Sox-9, aggrecan and collagen Ⅱ, which were chondrocyte specific, were upregulated in medium containing TGF-β1 under hypoxia (2% O2). Likewise, gene expression of HIF-1 a, which was characteristics of in- tervertebral discs, was also upregulated. Simultaneously, Alcian blue staining exhibited the formation of many GAGs. Conclusions: The approach in our experiment is a simple and effective way to acquire a large quantity of homogenous ADSCs. Rat ADSCs can be differentiated into nucleus pulposus-like cells. ADSCs may replace bone marrow mesenchymal stem cells as a new kind of seed cells in regeneration of degenerated intervertebral discs using cell transolantation therarw.展开更多
基金Supported by the Shandong Provincial Post-Doctoral Special Fund Innovative Project(No.S43201403)the Post-Doctoral Applied Research Project of Qingdao City(No.Q51201413)the Shandong Provincial Natural Science Foundation,China(No.ZR2015PD004)
文摘Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome background. Transcriptomic analysis based on high-throughput RNA sequencing has been shown to be an efficient and powerful method for the discovery of relevant genes in non-model and genome reference-free organisms. Here, we constructed two c DNA libraries(control and Cd exposure groups) from the digestive gland of C. farreri and compared the transcriptomic data between them. A total of 227 673 transcripts were assembled into 105 071 unigenes, most of which shared high similarity with sequences in the NCBI non-redundant protein database. For functional classification, 24 493 unigenes were assigned to Gene Ontology terms. Additionally, Eu Karyotic Ortholog Groups and Kyoto Encyclopedia of Genes and Genomes analyses assigned 12 028 unigenes to 26 categories and 7 849 unigenes to five pathways, respectively. Comparative transcriptomics analysis identified 3 800 unigenes that were differentially expressed in the Cd-treated group compared with the control group. Among them, genes associated with heavy metal accumulation were screened, including metallothionein, divalent metal transporter, and metal tolerance protein. The functional genes and predicted pathways identified in our study will contribute to a better understanding of the metabolic and immune system in the digestive gland of C. farreri. In addition, the transcriptomic data will provide a comprehensive resource that may contribute to the understanding of molecular mechanisms that respond to marine pollutants in bivalves.
基金Supported by The Natural Science Foundation of Anhui Province,No. 090413118
文摘AIM: To investigate the expression and significance of caudal-related homeobox transcription factor (Cdx2) in gastric carcinoma (GC) and precancerous lesions. METHODS: The expression of Cdx2 in GC, precancer- ous lesions and normal gastric mucosa were detected using immunohistochemical method. Hematoxylin and eosin staining, alcian blue/periodic acid-schiff and high iron diamine/alcian blue staining were used to classify intestinal metaplasia (IM) and GC. RESULTS: Cdx2 was not detected in normal gas- tric mucosa. Cdx2 expression was detected in 87.1% (101/116) of IM, 50% (36/72) of dysplasia and 48.2% (41/85) of GC. The Cdx2-expressing cells in IM were more prevalent than in dysplasia and carcinoma (P 〈 0.05). There was no relationship between Cdx2 ex- pression and the classification of IM or the degree of dysplasia. Expression of Cdx2 was significantly higher in intestinal-type carcinoma than in diffuse and mixed- type carcinoma (P 〈 0.05). Positive expression of Cdx2was mainly found in moderately to well differentiated GC. There was a negative association between nuclear Cdx2 expression and lymph node metastasis and tumor, nodes, metastasis stage of GC (P 〈 0.05). The patients with Cdx2-positive expression showed a higher survival rate than those with Cdx2-negative expression (P = 0.038). Multivariate analysis revealed that the expres- sion of Cdx2 and lymph node metastasis were indepen- dent prognostic indicators of GC (P 〈 0.05).
文摘OBJECTIVE To investigate the correlations between the expression of thyroid transcription factor-1 (TTF-1) and apoptosis and angiogenesis in lung carcinomas. METHODS A 829 microarray of the paraffin tissue chips was constructed, which contained 196 lung carcinomas, 10 normal lung tissues, and 1 muscular tissue. Terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) and immunohistochemical SP method were used to detect apoptosis and expression of TTF-1 and CD34 in different types of lung carcinomas. A Leica Q500 MC image analysis system was used to measure and calculate TTF-1 positive unit (PU), apoptotic index (AI) and microvessel density (MVD). RESULTS AI of lung small cell carcinoma and large cell carcinoma were smaller than those of lung adenocarcinoma and squamous cell carcinoma (P = 0.000). AI of lung carcinomas with lymph node metastases was smaller than that of those without (P = 0.039). AI of lung carcinomas in TNM stage I-IV was smaller than that in stage I (P = 0.008). The PU of the TTF-1 was negatively correlated with AI in small cell lung carcinoma (r = -0.752, P = 0.000). MVD of lung carcinomas without lymph node metastases was smaller than that of those with lymph node metastasis (P = 0.031). MVD of lung carcinomas in TNM stage I was smaller than that in stage I-IV (P = 0.040). The PU of TTF-1 was positively correlated with MVD in lung adenocarcinoma (r = 0.708, P = 0.000). CONCLUSION There is a negative correlation between TTF-1 PU and AI in small cell lung carcinoma. TTF-1 PU and AI may be correlated with each other. There is a positive correlation between TTF-1 PU and MVD in lung adenocarcinoma. TTF-1 may induce the development of lung adenocarcinoma by inducing tumor angiogenesis.
基金supported by grants to Zeng Qing Ping from National Natural Science Foundation of China(81273620)
文摘Calorie restriction(CR) promotes longevity among distinct organisms from yeast to mammals. Although CR-prolonged lifespan is believed to associate with enhanced respiratory activity, it is apparently controversial for accelerated energy consumption regardless of insufficient nutrient intake. In reconciling the contradiction of less food supply versus much metabolite dispense, we revealed a CR-based mode of dual-phase responses that encompass a phase of mitochondrial enhancement(ME) and a phase of post-mitochondrial enhancement(PME), which can be distinguished by the expression patterns and activity dynamics of mitochondrial signatures. ME is characterized by global antioxidative activation, and PME is denoted by systemic metabolic modulation. CR-mediated aging-delaying effects are replicated by artesunate, a semi-synthetic derivative of the antimalarial artemisinin that can alkylate heme-containing proteins, suggesting artesunate-heme conjugation functionally resembles nitric oxide-heme interaction. A correlation of artesunate-heme conjugation with cytochrome c oxidase activation has been established from adduct formation and activity alteration. Exogenous hydrogen peroxide also mimics CR to trigger antioxidant responses, affect signaling cascades, and alter respiratory rhythms, implying hydrogen peroxide is engaged in lifespan extension. Conclusively, artesunate mimics CR-triggered nitric oxide and hydrogen peroxide to induce antioxidative networks for scavenging reactive oxygen species and mitigating oxidative stress, thereby directing metabolic conversion from anabolism to catabolism, maintaining essential metabolic functionality, and extending life expectancy in yeast.
文摘Objective: To differentiate rat adipose tissue-derived mesenchymal stem cells (ADSCs) into cells with a nucleus pulposus-like phenotype in vitro, so as to lay a foundation for the cell-based transplantation therapy of degenerated intervertebral discs. Methods: Rat ADSCs were isolated only from the subcutaneous inguinal region and purified by limited dilution. ADSCs of the third passages were analyzed by fluorescence activated cell sorter (FACS) to detect the cell surface markers (Sca-1, CD44, CD45, CDI lb). To induce ADSCs to- wards a nucleus pulposus-like phenotype, ADSCs were immobilized in 3-dimensional alginate hydrogels and cultured in an inducing medium containing transforming growth factor-beta1 (TGF- β1) under hypoxia (2% O2), while control groups under normoxia (21% O2) in alginate beads in medium with or without the presence of TGF-β 1. Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) was carried out to evaluate phenotypic and biosynthetic activities in the process of differentiation. Meanwhile, Alcian blue staining were used to detect the formation of sulfated glycosaminoglycans (GAGs) in the differentiated cells. Results: The purified ADSCs were fibroblast-like and proliferated rapidly in vitro. The flow cytometry showed that ADSCs were positive for Sca-1 and CD44, negative for CD45 and CD11b. The results of RT-PCR manifested that the gene expressions of Sox-9, aggrecan and collagen Ⅱ, which were chondrocyte specific, were upregulated in medium containing TGF-β1 under hypoxia (2% O2). Likewise, gene expression of HIF-1 a, which was characteristics of in- tervertebral discs, was also upregulated. Simultaneously, Alcian blue staining exhibited the formation of many GAGs. Conclusions: The approach in our experiment is a simple and effective way to acquire a large quantity of homogenous ADSCs. Rat ADSCs can be differentiated into nucleus pulposus-like cells. ADSCs may replace bone marrow mesenchymal stem cells as a new kind of seed cells in regeneration of degenerated intervertebral discs using cell transolantation therarw.