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Published online: December 28, 2015 (DOI: 10.4238/2015.December.28.19). Corrected after publication: June 24, 2016 (DOI: 10.4238/gmr.150267961). There is a change in the corresponding author, who should be Y.F. Liu.

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In this study, a dynamic three-dimensional cell culture technology was used to expand and differentiate rat pancreatic duct-derived stem cells (PDSCs) into islet-like cell clusters that can secrete insulin. PDSCs were isolated from rat pancreatic tissues by in situ collagenase digestion and density gradient centrifugation. Using a dynamic three-dimensional culture technique, the cells were expanded and differentiated into functional islet-like cell clusters, which were characterized by morphological and phenotype analyses. After maintaining 1 x 108 isolated rat PDSCs in a dynamic three-dimensional cell culture for 7 days, 1.5 x 109 cells could be harvested. Passaged PDSCs expressed markers of pancreatic endocrine progenitors, including CD29 (86.17%), CD73 (90.73%), CD90 (84.13%), CD105 (78.28%), and Pdx-1. Following 14 additional days of culture in serum-free medium with nicotinamide, keratinocyte growth factor (KGF), and b fibroblast growth factor (FGF), the cells were differentiated into islet-like cell clusters (ICCs). The ICC morphology reflected that of fused cell clusters. During the late stage of differentiation, representative clusters were non-adherent and expressed insulin indicated by dithizone (DTZ)-positive staining. Insulin was detected in the extracellular fluid and cytoplasm of ICCs after 14 days of differentiation. Additionally, insulin levels were significantly higher at this time compared with the levels exhibited by PDSCs before differentiation (P < 0.01). By using a dynamic three-dimensional cell culture system, PDSCs can be expanded in vitro and can differentiate into functional islet-like cell clusters.

Assuntos

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The prevalence rates of anti-citrullinated protein/peptide antibodies (ACPAs) were investigated in a cohort of juvenile idiopathic arthritis (JIA) patients, and their diagnostic performances were compared. ACPAs, including anti-cyclic citrullinated peptide IgG (anti-CCP), anti-CCP IgG/IgA (anti-CCP3.1), citrullinated recombinant rat filaggrin antibodies (CPA), anti-mutated citrullinated vimentin (anti-MCV), and antibodies to citrullinated human IgG-derived peptides (RA/CP), were measured in the sera from 81 JIA patients. Serum samples from 55 children with other joint diseases or viral infections and 49 healthy donors were tested as controls. Of the 81 JIA patients, 7 (8.6%), 8 (9.9%), 17 (21.0%), 23 (28.4%), and 18 (22.2%) were found to be positive for anti-CCP, anti-CCP3.1, CPA, anti-MCV, and RA/CP, respectively, with specificities of 98.1, 95.1, 93.3, 84.6, and 86.5%. Analysis by subtype revealed that 7/7 (100%) of RF-positive polyarticular JIA patients tested positive at high serum levels for anti-MCV or RA/CP, and 5/7 (71.4%) were positive for anti-CCP, anti- CCP3.1, or CPA (P < 0.001, compared with controls). Eighteen of 81 JIA patients demonstrated joint erosions on radiographs and erosive arthritis occurred more often in ACPAs positive patients (P < 0.01). Our findings indicate that although ACPAs are not satisfactory screening biomarkers for JIA due to low sensitivity, ACPA measurement can aid in diagnosing RF-positive polyarticular JIA and identifying JIA patients with severe bone involvement. The diagnostic performance of each ACPA in JIA is different, and the careful selection of assays is necessary.

Assuntos

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MicroRNA-452 (miR-452) is dysregulated in some human malignancies, and is correlated with tumor progression. However, its expression and function in human colorectal cancer (CRC) remain unclear. The aim of our study was to explore the effects of miR-452 in CRC tumorigenesis and development. Using reverse transcription quantitative real-time polymerase chain reaction, we detected miR-452 expression in CRC cell lines and primary tumor tissues. We also examined the association between miR-452 expression and clinicopathological factors. We then investigated the effects of miR-452 on the biological behavior of CRC cells. miR-452 expression was significantly downregulated in CRC compared with the adjacent noncancerous tissues. A low level of miR-452 was associated with larger tumor size, deeper invasion depth, and advanced TNM stage. Multivariate Cox regression analysis identified decreased miR-452 expression as an independent factor predicting poor prognosis for CRC patients. In addition, in vitro functional analysis showed that overexpression of miR-452 in HCT116 cells reduced cell proliferation, promoted cell apoptosis, and inhibited cell invasion and migration. These findings indicate that miR-452acts as a tumor suppressor in CRC, and would serve as a novel molecular therapeutic agent for the treatment of the disease.

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Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.(AU)

Assuntos

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Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.

Assuntos

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Despite years of effort, current therapies for diabetic wounds are still not fully efficacious. Emerging evidence has suggested that microRNAs (miRNAs) play key roles in multiple physiological and pathological processes in eukaryotes, and could potentially be powerful therapeutic tools. This study investigated the differential expression profiling of miRNAs in cutaneous wounds in streptozotocin-induced diabetic rats and normal rats, and its significance in diabetic wound healing. Using microarrays, 18 miRNAs were identified as being upregulated and 65 as being downregulated in the diabetic group. The miRNA profiling results were validated by quantitative reverse transcriptase polymerase chain reaction. Finally, functional annotation analysis using the DAVID and miR2Subpath databases revealed that the differentially expressed miRNAs were involved in MAPK signaling pathways, the Wnt signaling pathway, and other signaling pathways that may be closely linked to wound healing. This study provides an experimental foundation for further investigation of mechanisms that underlie poor diabetic wound healing, and of miRNA-based therapies that are associated with wound healing.

Assuntos

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Tomato yellow leaf curl virus is one of the main diseases affecting tomato production worldwide. Previous studies have shown that Ty-2 is an important resistance gene located between molecular markers C2_At2g28250 (82.3 cM) and T0302 (89.0 cM), and exhibits strong resistance to tomato yellow leaf curl virus in Asia. In this study, Ty-2 candidate genes were subjected to bioinformatic analysis for the sequenced tomato genome. We identified 69 genes between molecular markers C2_At2g28250 and T0302, 22 of which were disease-related resistant genes, including nucleotide binding site-leucine-rich repeat disease resistance genes, protease genes (protein kinase, kinase receptor, and protein isomerase), cytochromes, and transcription factors. Expressed sequence tag analysis revealed that 77.3% (17/22) of candidate disease-resistance genes were expressed, involving 143 expressed sequence tags. Based on full-length cDNA sequence analysis, 7 candidate genes were found, 4 of which were involved in tomato responses to pathogens. Microarray expression analysis also showed that most candidate genes were involved in the tomato responses to multiple pathogens, including fungi, viruses, and bacteria. RNA-seq expression analysis revealed that all candidate genes participated in tomato growth and development.

Assuntos

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This study aimed to discuss the effects of 3 different analgesia methods on serum IL-6 and IL-10 in patients after cesarean delivery. Thirty full-term women, who underwent cesarean delivery, were randomly assigned to 3 analgesia groups (10 cases each) as follows: intramuscular injection of 100 mg pethidine (NC group), patient controlled epidural analgesia (PCEA) of 5 mg morphine plus 150 mg ropivacaine (MR group), and patient controlled intravenous analgesia (PCIA) of 150 mg sufentanil plus 5 mg droperidol (SF group). An electronic analgesia pump was available in all 3 groups. At 4, 12, 24, and 48 h after surgery, visual analogue scale (VAS) pain scores were evaluated, IL-6 and IL-10 serum levels were measured, and adverse reactions were documented. The MR and SF groups responded well to analgesia. VAS scores at 12 and 24 h in these 2 groups were significantly lower than those in the NC group (P < 0.05). IL-6 and IL-10 levels were elevated to varying degrees postoperatively in all 3 groups. In the MR and SF groups, no significant difference occurred at each time point (P > 0.05), but compared with the NC group, significant differences were observed at 12 and 24 h (P < 0.05). Both PCIA and PCEA produced good analgesic effect, decreased postoperative level of serum IL-6, promoted release of anti-inflammatory factor IL-10, maintained balance in postoperative serum IL-6 level, and reduced the postoperative inflammatory response. Adverse reactions were significantly higher with epidural morphine than with intravenous sufentanil.

Assuntos

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Ischemia time during transplantation has greatly restricted the quality and utilization of grafts. To improve the quality of islet transplantation, adenosine was added into the University of Wisconsin (UW) pancreas perfusate to assess its effect on islet yield and function in porcine pancreas. Ten pancreata from donation after cardiac death pigs were obtained and randomly divided into two groups: control group (N = 5) with UW perfusion solution, and experimental group (N = 5) with adenosine-enriched UW perfusion solution. The yield and purity of the islet cells were counted after they were collected, purified, and stained with dithizone. Acridine orange/propidium iodide staining was applied to determine islet cell viability. Islet function was assessed by glucose stimulated insulin secretion assays, and released insulin was quantified by enzyme-linked immunosorbent assay. The metabolic substrates of the pancreas were analyzed by trace dialysis technology. We found that the addition of adenosine in UW perfusion solution significantly increased the yield, purity and viability of islet cells, as well as enhanced their insulin release. In addition, the levels of metabolic substrates, pyruvate and lactate, were significantly reduced. The addition of adenosine could effectively increase islet cell viability during mechanical perfusions, which may improve islet transplantation. This perfusion protocol may be clinically feasible, and should be considered in the clinical setting.

Assuntos