Panax notoginseng stems and leaves affect microbial community and function in cecum of duzang pigs.

Panax notoginseng is a Chinese medicine with a long history in which stems and leaves are the wastes of processing Panax notoginseng and have not been effectively utilized. The effects of diets containing Panax notoginseng stems and leaves on the cecal short-chain fatty acid (SCFA) concentration and microbiome of independent pigs were studied. Diets containing Panax notoginseng stems and leaves did not affect the concentration of SCFA in the cecal contents of Duzang pigs but affected the microbial composition and diversity. Firmicutes, Proteobacteria, and Bacteroidetes dominate in the cecal of Duzang pigs. Feeding Duzang pigs with a 10% Panax notoginseng stems and leaves diet increases the abundance of Lactobacillus, Christensenellaceae R-7 group, and Akkermansia in the cecal. We found 14 genera positively associated with acetate, and they were Lactobacillus, Ruminococcaceae UCG 005, Ruminiclostridium 6; Escherichia Shigella and Family XIII AD3011 group showed negative correlations. Solobacterium, Desulfovibrio, and Erysipelatoclostridium were positively associated with propionate. Campylobacter, Clostridium sensu stricto 11, and Angelakisella were positively associated with butyrate. In conclusion, Panax notoginseng stems and leaves could affect the cecal microbial community and functional composition of Duzang pigs. Panax notoginseng stems and leaves reduce the enrichment of lipopolysaccharide biosynthetic pathway of the cecal microbiome, which may have a positive effect on intestinal health. The higher abundance of GH25 family in Duzang pig's cecal microbiome of fed Panax notoginseng stems and leaves diet. This increase may be the reason for the microbial diversity decrease.

Single-nucleus transcriptome sequencing reveals hepatic cell atlas in pigs.

BACKGROUND: As the largest substantive organ of animals, the liver plays an essential role in the physiological processes of digestive metabolism and immune defense. However, the cellular composition of the pig liver remains poorly understood. This investigation used single-nucleus RNA sequencing technology to identify cell types from liver tissues of pigs, providing a theoretical basis for further investigating liver cell types in pigs. RESULTS: The analysis revealed 13 cells clusters which were further identified 7 cell types including endothelial cells, T cells, hepatocytes, Kupffer cells, stellate cells, B cells, and cholangiocytes. The dominant cell types were endothelial cells, T cells and hepatocytes in the liver tissue of Dahe pigs and Dahe black pigs, which accounts for about 85.76% and 82.74%, respectively. The number of endothelial cells was higher in the liver tissue of Dahe pigs compared to Dahe black pigs, while the opposite tendency was observed for T cells. Moreover, functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic endothelial cells were significantly enriched in the protein processing in endoplasmic reticulum, MAPK signaling pathway, and FoxO signaling pathway. Functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic T cells were significantly enriched in the thyroid hormone signaling pathway, B cell receptor signaling pathway, and focal adhesion. Functional enrichment analysis demonstrated that the differentially expressed genes in pig hepatic hepatocytes were significantly enriched in the metabolic pathways. CONCLUSIONS: In summary, this study provides a comprehensive cell atlas of porcine hepatic tissue. The number, gene expression level and functional characteristics of each cell type in pig liver tissue varied between breeds.

Comparison of differentially expressed genes in longissimus dorsi muscle of Diannan small ears, Wujin and landrace pigs using RNA-seq.

Introduction: Pig growth is an important economic trait that involves the co-regulation of multiple genes and related signaling pathways. High-throughput sequencing has become a powerful technology for establishing the transcriptome profiles and can be used to screen genome-wide differentially expressed genes (DEGs). In order to elucidate the molecular mechanism underlying muscle growth, this study adopted RNA sequencing (RNA-seq) to identify and compare DEGs at the genetic level in the longissimus dorsi muscle (LDM) between two indigenous Chinese pig breeds (Diannan small ears [DSE] pig and Wujin pig [WJ]) and one introduced pig breed (Landrace pig [LP]). Methods: Animals under study were from two Chinese indigenous pig breeds (DSE pig, n = 3; WJ pig, n = 3) and one introduced pig breed (LP, n = 3) were used for RNA sequencing (RNA-seq) to identify and compare the expression levels of DEGs in the LDM. Then, functional annotation, Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and Protein-Protein Interaction (PPI) network analysis were performed on these DEGs. Then, functional annotation, Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and Protein-Protein Interaction (PPI) network analysis were performed on these DEGs. Results: The results revealed that for the DSE, WJ, and LP libraries, more than 66, 65, and 71 million clean reads were generated by transcriptome sequencing, respectively. A total of 11,213 genes were identified in the LDM tissue of these pig breeds, of which 7,127 were co-expressed in the muscle tissue of the three samples. In total, 441 and 339 DEGs were identified between DSE vs. WJ and LP vs. DSE in the study, with 254, 193 up-regulated genes and 187, 193 down-regulated genes in DSE compared to WJ and LP. GO analysis and KEGG signaling pathway analysis showed that DEGs are significantly related to contractile fiber, sarcolemma, and dystrophin-associated glycoprotein complex, myofibril, sarcolemma, and myosin II complex, Glycolysis/Gluconeogenesis, Propanoate metabolism, and Pyruvate metabolism, etc. In combination with functional annotation of DEGs, key genes such as ENO3 and JUN were identified by PPI network analysis. Discussion: In conclusion, the present study revealed key genes including DES, FLNC, PSMD1, PSMD6, PSME4, PSMB4, RPL11, RPL13A, ROS23, RPS29, MYH1, MYL9, MYL12B, TPM1, TPM4, ENO3, PGK1, PKM2, GPI, and the unannotated new gene ENSSSCG00000020769 and related signaling pathways that influence the difference in muscle growth and could provide a theoretical basis for improving pig muscle growth traits in the future.

Effects of substitution of soybean meal with rapeseed meal and glutamine supplementation on growth performance, intestinal morphology, and intestinal mucosa barrier of Qiandongnan Xiaoxiang Chicken.

OBJECTIVE: The present study was to evaluate the effects of different rapeseed meal substitution (RSM) and glutamine (Gln) supplementation on growth performance, intestine morphology, and intestinal mucosa barrier of broilers. METHODS: Four hundred and twenty Qiandongnan Xiaoxiang Chicken at 1 day of age with similar weight were chosen and were randomly assigned into 7 groups, consisting of 10 replicates per group and 6 broilers per replicate. Three groups were provided with diets separately containing 0%, 10%, and 20% RSM, and the other four groups were fed with diets separately supplemented with 0.5% and 1% Gln based on the inclusion of 10% and 20% RSM. At 21 and 42 days of age, 10 broilers per group were chosen to collect plasma and intestinal samples for further analysis. RESULTS: The results showed that 10% RSM decreased average daily feed intake (ADFI) and average daily weight gain (ADG) of broilers at 21 days of age (p<0.05). Furthermore, both ADFI and ADG of broilers at 21 and 42 days of age were decreased by 20% RSM, while feed conversion ratio (FCR) was increased (p<0.05). Besides, 10% RSM resulted in lower intestinal villus height and the ratio of villus height to crypt depth, deeper crypt depth (p<0.05), combined with the lower mRNA expressions of occludin, claudin-1, and zonula occludens-1 (ZO-1) in broilers at 21 days of age (p<0.05). Similar results were also observed in broilers at 21 and 42 days of age fed with 20% RSM. However, 1% Gln improved the growth performance of broilers fed with 10% and 20% RSM (p<0.05), ameliorated intestine morphology and elevated mRNA expressions of occludin, claudin-1 and ZO-1 (p<0.05). CONCLUSION: In conclusion, the increasing inclusion of RSM resulted in more serious effects on broilers, however, 1.0% Gln could reverse the negative effects induced by the inclusion of RSM.

Dietary guanidineacetic acid supplementation ameliorated meat quality and regulated muscle metabolism of broilers subjected to pre-slaughter transport stress by metabolomics analysis.

Pre-slaughter transport stress could induce multiple comprehensive variations in physiological and metabolic parameters of broilers. However, the entire metabolomics of pre-slaughter transport stress and supplementation of exogenous energy regulatory substances on broilers is still poorly understood. The metabolome characteristics of broilers subjected to 3 h pre-slaughter transport stress combined with 1,200 mg/kg guanidinoacetic acid (GAA1,200) supplementation were analyzed using gas chromatography-mass spectrometry (GC-MS) in this study. The results showed that, compared to the control group (no transport), 3 h pre-slaughter transport stress (T3h) decreased creatine (Cr), phosphocreatine (PCr) and adenosine triphosphate (ATP), and increased adenosine diphosphate (ADP), adenosine monophosphate (AMP) and the ratio of AMP to ATP in pectoralis muscle (PM) of broilers by high performance liquid chromatography (HPLC) analysis. However, GAA1,200 supplementation reversed the negative effects induced by 3 h pre-slaughter transport stress. Besides, GAA1,200 supplementation elevated mRNA expression of creatine transporter in PM. Our metabolomics approaches demonstrated that 38 and 48 significant metabolites were separately identified between the control group and T3h group, and T3h group and 3 h pre-slaughter transport stress combined with GAA1,200 supplementation group using the standard of variable importance in the projection values >1 and P < 0.05. Among these, the metabolites involved in amino acid metabolism (alanine, glycine, serine, threonine, cysteine , methionine, phenylalanine, tyrosine, and tryptophan), oxidative stress (3-methylhistidine, 1-methylhistidine and glutathione), non-protein amino acid (citrulline) metabolism, and energy metabolism (Cr, PCr, sarcosine, and glycocyamine) were confirmed through pathway enrichment analysis, which could be chosen as suitable candidate targets for further analysis of the effects of exogenous energy substances on broilers subjected to transport stress.

Optogenetics stimulates nerve reorganization in the contralesional anterolateral primary motor cortex in a mouse model of ischemic stroke.

The anterolateral motor cortex of rodents is an important motor auxiliary area, and its function is similar to that of the premotor area in humans. Activation and inhibition of the contralesional anterolateral motor cortex (cALM) have been shown to have direct effects on motor behavior. However, the significance of cALM activation and inhibition in the treatment of stroke remains unclear. This study investigated the role of optogenetic cALM stimulation in a mouse model of cerebral stroke. The results showed that 21-day optogenetic cALM inhibition, but not activation, improved neurological function. In addition, optogenetic cALM stimulation substantially altered dendritic structural reorganization and dendritic spine plasticity, as optogenetic cALM inhibition resulted in increased dendritic length, number of dendritic spines, and number of perforated synapses, whereas optogenetic activation led to an increase in the number of multiple synapse boutons and the number of dendritic intersections. Furthermore, RNA-seq analysis showed that multiple biological processes regulated by the cALM were upregulated immediately after optogenetic cALM inhibition, and that several immediate-early genes (including cFOS, Erg1, and Sema3f) were expressed at higher levels after optogenetic inhibition than after optogenetic activation. These results were confirmed by quantitative reverse transcription-polymerase chain reaction. Finally, immunofluorescence analysis showed that the c-FOS signal in layer V of the primary motor cortex in the ischemic hemisphere was higher after optogenetic cALM activation than it was after optogenetic cALM inhibition. Taken together, these findings suggest that optogenetic cALM stimulation promotes neural reorganization in the primary motor cortex of the ischemic hemisphere, and that optogenetic cALM inhibition and activation have different effects on neural plasticity. The study was approved by the Experimental Animal Ethics Committee of Fudan University (approval No. 201802173S) on March 3, 2018.

Metabolic Engineering of Non-carotenoid-Producing Yeast Yarrowia lipolytica for the Biosynthesis of Zeaxanthin.

Zeaxanthin is vital to human health; thus, its production has received much attention, and it is also an essential precursor for the biosynthesis of other critical carotenoids such as astaxanthin and crocetin. Yarrowia lipolytica is one of the most intensively studied non-conventional yeasts and has been genetically engineered as a cell factory to produce carotenoids such as lycopene and ß-carotene. However, zeaxanthin production by Y. lipolytica has not been well investigated. To fill this gap, ß-carotene biosynthesis pathway has been first constructed in this study by the expression of genes, including crtE, crtB, crtI, and carRP. Three crtZ genes encoding ß-carotene hydroxylase from different organisms were individually introduced into ß-carotene-producing Y. lipolytica to evaluate their performance for producing zeaxanthin. The expression of crtZ from the bacterium Pantoea ananatis (formerly Erwinia uredovora, Eu-crtZ) resulted in the highest zeaxanthin titer and content on the basis of dry cell weight (DCW). After verifying the function of Eu-crtZ for producing zeaxanthin, the high-copy-number integration into the ribosomal DNA of Y. lipolytica led to a 4.02-fold increase in the titer of zeaxanthin and a 721% increase in the content of zeaxanthin. The highest zeaxanthin titer achieved 21.98 ± 1.80 mg/L by the strain grown on a yeast extract peptone dextrose (YPD)-rich medium. In contrast, the highest content of DCW reached 3.20 ± 0.11 mg/g using a synthetic yeast nitrogen base (YNB) medium to culture the cells. Over 18.0 g/L of citric acid was detected in the supernatant of the YPD medium at the end of cultivation. Furthermore, the zeaxanthin-producing strains still accumulated a large amount of lycopene and ß-carotene. The results demonstrated the potential of a cell factory for zeaxanthin biosynthesis and opened up an avenue to engineer this host for the overproduction of carotenoids.

Transcriptional profiling of buffalo mammary gland with different milk fat contents.

Milk fat is the most important energy substance in milk and contributes to its quality and health benefits. Water buffalo milk is well known for its high milk quality with higher fat contents compared with cattle milk. Dehong buffalo is a unique local swamp breed in Yunnan Province with higher milk fat and excellent milk quality which provides a good model for the investigation of the molecular mechanisms of milk fat deposition. In this study, we used RNA-seq to obtain mammary tissue transcriptomics of buffalo with different milk fat phenotypes including high(H), medium (M)and low (L) fat content groups. Comparative analyses of buffalo among three groups yielded differentially expressed genes (DEGs). Analyzing the number of different genes among H_VS_L, H_VS_M, and M_VS_L showed the same expression pattern between H_VS_M. The increasing expression levels of genes including CSN1S1, BTN1A1, LALBA, ALDH1L2, SCD and MUC15, and down-regulated expression levels of genes containing CCL2, CRABP2, ADTRP, CLU and C4A in H_VS_L and M_VS_L were found. GO and KEGG enriched pathways revealed these DEGs involved in milk protein and fat as well as immune response. The findings would enhance the understanding of the interplay between the milk composition and immune response, which suggests that the immune capacity should be considered when we tried to improve the milk quality.

Challenges and Potential in Increasing Lutein Content in Microalgae.

Research on enhancing lutein content in microalgae has made significant progress in recent years. However, strategies are needed to address the possible limitations of microalgae as practical lutein producers. The capacity of lutein sequestration may determine the upper limit of cellular lutein content. The preliminary estimation presented in this work suggests that the lutein sequestration capacity of the light-harvesting complex (LHC) of microalgae is most likely below 2% on the basis of dry cell weight (DCW). Due to its nature as a structural pigment, higher lutein content might interfere with the LHC in fulfilling photosynthetic functions. Storing lutein in a lipophilic environment is a mechanism for achieving high lutein content but several critical barriers must be overcome such as lutein degradation and access to lipid droplet to be stored through esterification. Understanding the mechanisms underlying lipid droplet biogenesis in chloroplasts, as well as carotenoid trafficking through chloroplast membranes and carotenoid esterification, may provide insight for new approaches to achieve high lutein contents in algae. In the meantime, building the machinery for esterification and sequestration of lutein and other hydroxyl-carotenoids in model microorganisms, such as yeast, with synthetic biology technology provides a promising option.

[Clinical efficacy of different rehabilitation modes for lumbar degenerative diseases after operation].

OBJECTIVE: To compare clinical effects of different postoperative rehabilitation modes on lumbar degenerative diseases, and explore influence of rehabilitation mode and other factors on postoperative effect. METHODS: From June 2013 to July 2016, totally 900 patients were admitted from nine tertiary hospitals in Beijing to perform single segment bone grafting and internal fixation due to lumbar degenerative diseases were prospectively analyzed. There were 428 males and 472 females, the age of patient over 18 years old, with an average of (51.42±12.41) years old;according to patients' subjective wishes and actual residence conditions, all patients were divided into three groups, named as observation group 1 (performed integrated rehabilitation approach and orthopedic treatment model intervention), observation group 2 (performed integrated rehabilitation approach and orthopedic treatment, classified rehabilitation model intervention), and control group(performed routine rehabilitation model intervention). Visual analogue scale(VAS), Oswestry Disability Index(ODI) and Japanese Orthopaedic Association (JOA) were used to evaluate postoperative efficacy among three groups at 24 weeks. Possible factors affecting the postoperative efficacy including age, age grouping, gender, body mass index (BMI), BMI grouping, education level, visiting hospital, payment method of medical expenses, preoperative complications, preoperative JOA score, clinical diagnosis, surgery section, operative method, intraoperative bleeding volume, postoperative complications and rehabilitation mode were listed as independent variables, and postoperative ODI score at 24 weeks as dependent variables. Univariate analysis was used to analyze relationship between influencing factors and postoperative efficacy. Multiple linear regression was used to analyze relationship between influencing factors, rehabilitation mode and postoperative ODI score at 24 weeks, in further to find out the main reasons which affect postoperative efficacy, and to analyze impact of rehabilitation mode on postoperative efficacy. RESULTS: All patients were followed up for 24 weeks after operation. All incisions healed at stage I with stable internal fixation. (1)Evaluation of postoperative efficacy:① There were no statistical differences in preoperative VAS and ODI among three groups(P>0.05), the degree of pain and dysfunction decreased among three groups after operation, and had differences in postoperative VAS and ODI among three groups (P<0.05). There were no significant differences between observation group 1 and observation group 2(P>0.05); while compared with observation group 1 and control group, observation group 2 and control group, there were significant differences (P<0.05). ②The function among three groups were improved in varying degrees after operation. There was difference in JOA score among three groups before operation and 24 weeks after operation (P<0.05). There were no difference in JOA score among three groups between observation group 1 and observation group 2 (P>0.05);while compared with observation group 1 and control group, observation group 2 and control group, there were significant differences (P<0.05). (2)Influencing factors at 24 weeks after operation:①Univariate analysis showed gender, age, age grouping, education level, preoperative complications, clinical diagnosis, operative section, operative method, preoperative JOA score and rehabilitation mode had statistical significance with postoperative ODI score at 24 weeks (P<0.05). BMI, BMI grouping, payment method of medical expenses, visiting hospital, intraoperative bleeding volume, postoperative complications had no statistical significance with postoperative ODI score at 24 weeks (P<0.05).②Multivariate analysis results showed gender, rehabilitation mode, age, preoperative JOA score entered the equation eventually, stepwise multiple linear equation obtained had statistical significance (F=12.294, P= 0.000). Among rehabilitation mode, standardized regression coefficient of the integrated rehabilitation approach and orthopedic treatment with classified rehabilitation model was absolute value of the largest (0.176), which had the greatest influence on postoperative curative effect. The degree of dysfunction in control group was higher than that in observation group 1 and observation group 2. Postoperative dysfunction was more severe in males than that of in females. Older age has higher degree of dysfunction after operation. Lower preoperative JOA score has higher degree of dysfunction after operation. CONCLUSION: Preoperative JOA score, gender, age could predict postoperative clinical effects of lumbar degenerative diseases in varying degrees treated with single level bone graft fusion and internal fixation. Different rehabilitation modes could improve clinical effects. Intergrated rehabilitation orthopedic treatment model and integrated rehabilitation approach and orthopedic treatment with classifiedrehabilitation model are superior to conventional rehabilitation model in improving patients' postoperative function and relieving pain, which is worthy of promoting in clinical.

Plant-Derived Polysaccharides Regulated Immune Status, Gut Health and Microbiota of Broilers: A Review.

In modern intensive breeding system, broilers are exposed to various challenges, such as diet changes and pathological environment, which may cause the increase in the incidence rate and even death. It is necessary to take measures to prevent diseases and maintain optimal health and productivity of broilers. With the forbidden use of antibiotics in animal feed, polysaccharides from plants have attracted much attention owing to their lower toxicity, lower drug resistance, fewer side effects, and broad-spectrum antibacterial activity. It had been demonstrated that polysaccharides derived from plant exerted various functions, such as growth promotion, anti-inflammation, maintaining the integrity of intestinal mucosa, and regulation of intestinal microbiota. Therefore, the current review aimed to provide an overview of the recent advances in the impacts of plant-derived polysaccharides on anti-inflammation, gut health, and intestinal microbiota community of broilers in order to provide a reference for further study on maintaining the integrity of intestinal structure and function, and the related mechanism involved in the polysaccharide administration intervention.

Guanidino-Acetic Acid: A Scarce Substance in Biomass That Can Regulate Postmortem Meat Glycolysis of Broilers Subjected to Pre-slaughter Transportation.

The different substances in biomass can regulate the metabolism and reproduction of broilers. Guanidino-acetic acid (GAA) is a natural feed additive that showed a potential application in dietary for broilers, while its amount is scarce in biomass. The objective of the present study was to investigate the effects of dietary supplemented with GAA on muscle glycolysis of broilers subjected to pre-slaughter transportation. A total of 160 Qiandongnan Xiaoxiang chickens were randomly assigned into three treatments, including a basal control diet without GAA supplementation (80 birds) or supplemented with 600 mg/kg (40 birds) or 1,200 mg/kg (40 birds) GAA for 14 days. At the end of the experiment, the control group was equally divided into two groups, thus resulting in four groups. All birds in the four groups aforementioned were separately treated according to the following protocols: (1) no transport of birds of the control group fed with the basal diet; (2) a 3-h transport of birds of the control group fed with the basal diet; (3) a 3-h transport of birds fed with diets supplemented with 600 mg/kg GAA; and (4) a 3-h transport of birds fed with diets supplemented with 1,200 mg/kg GAA. The results demonstrated that 3-h pre-slaughter transport stress increased corticosterone contents and lowered glucose contents in plasma (P < 0.05), decreased pH24 h (P < 0.05), and resulted in inferior meat quality evidenced by elevating the drip loss, cooking loss, and L∗ value (P < 0.05). Meanwhile, 3-h pre-slaughter transport stress decreased the contents of Cr and ATP in muscle (P < 0.05) and elevated the ratio of AMP:ATP and the glycolytic potential of muscle (P < 0.05). Moreover, 3-h pre-slaughter transport resulted in a significant elevation of mRNA expressions of LKB1 and AMPKα2 (P < 0.05), as well as the increase in protein abundances of LKB1 phosphorylation and AMPKα phosphorylation (P < 0.05). However, 1,200 mg/kg GAA supplementation alleviated negative parameters in plasma, improved meat quality, and ameliorated postmortem glycolysis and energy metabolism through regulating the creatine-phosphocreatine cycle and key factors of AMPK signaling. In conclusion, dietary supplementation with 1,200 mg/kg GAA contributed to improving meat quality via ameliorating muscle energy expenditure and delaying anaerobic glycolysis of broilers subjected to the 3-h pre-slaughter transport.

A hybrid classifier based on nonlinear-PCA and deep belief networks with applications in dysphagia diagnosis.

Traditional dysphagia prescreening diagnostic methods require doctors specialists to give patients a total score based on a water swallow test scale. This method is limited by the high dimensionality of the diagnostic elements in the water swallow test scale with heavy workload (Towards each patient, the scale requires the doctors give score for 18 diagnostic elements respectively) as well as the difficulties of extracting and using the diagnostic scale data's non-linear features and hidden expertise information (Even with the scale scores, specific diagnostic conclusions are still given by expert doctors under the expertise). In this paper, a hybrid classifier model based on Nonlinear-Principal Component Analysis (NPCA) and Deep Belief Networks (DBN) is proposed in order to effectively extract the diagnostic scale data's nonlinear features and hidden information and to provide the key scale elements' locating methods towards the diagnostic results (The key scale elements that affect different diagnostic conclusions are given to improve the efficiency and pertinence of diagnosis and reduce the workload of diagnosis). We demonstrate the effectiveness of the proposed method using the frame of 'information entropy theory'. Real dysphagia diagnosis examples from the China-Japanese Friendship Hospital are used to demonstrate applications of the proposed methods. The examples show satisfactory results compared to the traditional classifier.

ERK pathway activation contributes to the tumor-promoting effects of hepatic stellate cells in hepatocellular carcinoma.

BACKGROUND: Activated hepatic stellate cell (aHSC) play a critical role in hepatocellular carcinoma (HCC) progression crosstalking with cancer cell via various signaling pathways. The aim of our study is to explore the tumor-promoting effects of aHSCs on HCC via ERK pathway. METHODS: α-SMA, p-ERK and p-JNK expression levels in tumoral and peritumoral tissues of HCC were assessed by immunohistochemical and western blotting. The protein and mRNA expression levels in human hepatoma cell treated with aHSC conditioned medium (CM) were determined by western blotting and real-time quantitative PCR, respectively. Cell migration and invasion abilities were assessed using transwell assays. The proliferation ability of HCC cells induced by aHSCs-CM was detected by CCK-8 assay and cell cycle analysis. RESULTS: We found that aHSC number was positively correlated with p-ERK expression levels in tumoral tissues and aHSC-CM could time-dependently promote PCNA, p-ERK expression in HCC cells. Moreover, aHSC-CM enhanced HCC cells proliferation via ERK. Additionally, aHSC upregulated c-jun and cyclinD1 expression levels, accelerating the transition from G1 to the S phase of HCC cells, and this effect could be arrested by inhibiting ERK pathway. Furthermore, aHSC-CM promoted migration and invasion of HCC cells via ERK. Epithelial-mesenchymal transitions (EMT) phenomenon could be reversed by ERK suppression. CONCLUSION: High expression of p-ERK and aHSCs may be associated with the aggressive behavior of HCC cells. Secretions from aHSCs could promote proliferation and EMT of HCC cells via ERK1/2/c-jun/cyclinD1 axis or ERK pathway.

[Hepatic stellate cells inhibit the proliferation and invasion of HepG2 hepatocellular carcinoma via activating γδ T cells].

Objective To investigate the effects of γδ T cells co-cultured with human hepatic stellate cells (HSCs) on the proliferation and invasion of hepatocellular carcinoma (HCC) cells. Methods Peripheral γδ T cells of healthy volunteers and HCC patients were separated and collected by density gradient centrifugation and flow cytometry. TranswellTM chambers were used to co-culture LX-2 cells and γδ T cells. The concentration of IFN-γ in γδ T cell supernatant was detected by ELISA. The proliferation of HepG2 cells was analyzed by CCK-8 assay, and the invasion ability of HepG2 cells was evaluated by TranswellTM chamber assay. Results After co-cultured with LX-2 cells, the IFN-γ secretion in γδ T cells increased obviously, and the level of IFN-γ was higher in healthy volunteer group than in HCC patient group. The inhibitory effects of γδ T cells on the proliferation and invasion of HepG2 cells were enhanced after the co-culture, and the enhancement of inhibitory abilities was more obvious in healthy volunteer group than in HCC patient group. Conclusion HSC could activate γδ T cells and promote IFN-γ secretion of γδ T cells, thus enhancing the inhibitory effects of γδ T cells on the proliferation and invasion of hepatoma cells.

[Hepatic stellate cell conditioned medium induces proliferation and epithelial-mesenchymal transition via activating ERK1/2 signaling pathway in hepatoma cells].

Objective: To investigate the influence of hepatic stellate cells( HSCs) on malignant biological behavior of hepatoma cells and related mechanisms. Methods: Human hepatoma cell lines SMMC-7721 and Hep G2,and hepatic stellate cell line LX-2 were cultured separately. HSC conditioned medium( LX2-CM),MAPK specific inhibitor U0126 were used to treat hepatoma cells,separately or together. The invasion and migration abilities of hepatoma cells were detected by TranswellTMassay,and cell proliferation was analyzed by CCK-8 assay. The mRNA and protein expression levels of p-ERK1 /2,ERK1 /2,c-Myc,vimentin and E-cadherin were determined by real-time PCR and Western blot analysis,respectively. Results: LX2-CM promoted the proliferation,invasion and migration of SMMC-7721 and Hep G2 cells,and these effects were inhibited by U0126. LX2-CM up-regulated the expression levels of p-ERK1 /2,c-Myc,vimentin and down-regulated the expression level of E-cadherin. Conversely,after U0126 treatment,the expression levels of p-ERK1 /2,c-Myc and vimentin decreased significantly,while E-cadherin expression level increased. Conclusion: LX2-CM could activate c-Myc via ERK1 /2signaling pathway in hepatoma cells,and consequently promote cell proliferation,invasion and migration,and also induce epithelial-mesenchymal transition.

Selection of microalgae and cyanobacteria strains for bicarbonate-based integrated carbon capture and algae production system.

Using microalgae to capture CO2 from flue gas is an ideal way to reduce CO2 emission, but this is challenged by the high cost of carbon capture and transportation. To address this problem, a bicarbonate-based integrated carbon capture and algae production system (BICCAPS) has been proposed, in which bicarbonate is used for algae culture, and the regenerated carbonate from this process can be used to capture more CO2. High-concentration bicarbonate is obligate for the BICCAPS. Thus, different strains of microalgae and cyanobacteria were tested in this study for their capability to grow in high-concentration NaHCO3. The highest NaHCO3 concentrations they are tolerant to were determined as 0.30 M for Synechocystis sp. PCC6803, 0.60 M for Cyanothece sp., 0.10 M for Chlorella sorokiniana, 0.60 M for Dunaliella salina, and 0.30 M for Dunaliella viridis and Dunaliella primolecta. In further study, biomass production from culture of D. primolecta in an Erlenmeyer flask with either 0.30 M NaHCO3 or 2 % CO2 bubbling was compared, and no significant difference was detected. This indicates BICCAPS can reach the same biomass productivity as regular CO2 bubbling culture, and it is promising for future application.

The feasibility and validity of the comprehensive ICF core set for stroke in Chinese clinical settings.

OBJECTIVE: To inspect the feasibility and content validity of the comprehensive International Classification of Functioning, Disability and Health (ICF) core set for stroke by describing relevant aspects of functioning, disability and environmental factors affected in Chinese patients post stroke. DESIGN: Multicentre, cross-sectional study. SETTING: Department of rehabilitation medicine. SUBJECTS: The content validity was evaluated using frequency and percentage of 208 patients with a mean age of 60 years post stroke in China. OUTCOME MEASURES: Aspects of body function and structure, activity and participation, and environmental factors in the comprehensive ICF core set for stroke. RESULTS: Six ICF categories of body function were identified as a problem in over 90% (n = 187) of the patients (functions of the cardiovascular system and neuromusculoskeletal and movement-related functions). Impairments of brain, upper and lower extremity were identified as a problem in over 50% (n = 104) of the patients. Four ICF categories of activities and participation were documented as a problem in 100% (n = 208) of the patients (domestic and civic life). In environmental factors, nine ICF categories were documented as barriers by more than 10% (n = 20) and fewer than 50% (n = 104) of the patients (products and technology, physical geography, societal attitudes, services, systems and polices). Six ICF categories were identified as facilitators in over 90% (n = 187) of the patients (support and relationships and attitudes). CONCLUSION: The findings suggest that it is feasible to apply the comprehensive ICF core set for stroke in the Chinese clinical setting, after the appropriate reduction of some categories according to Chinese patients' characteristics and culture.