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Background & objectives: Swiss-type hereditary persistence of foetal haemoglobin (HPFH) has been shown to be responsible for the wide range of F cell levels in healthy Thai adults. However, a survey for F cells in healthy Thai adults has not been performed. This study was conducted to determine the F cell distribution in adult Thai blood donors and to assess the possible involvement of ?-thalassaemia and haemoglobin E (HbE) carriers in increased HbF levels. Methods: Thai blood donors (n=375, 205 males and 170 females) were included in the study. Blood samples were collected for measuring haemoglobin (Hb) concentration and haematocrit (Hct) and F cell levels. Hb and Hct levels were determined by automated blood counter, while F cells were quantified by flow cytometric analysis of F cells stained by fluorescein isothiocyanate-conjugated anti ?-globin monoclonal antibody. Finally, F cell levels were compared between blood samples having mean corpuscular volume (MCV ) <80 fl and ?80 fl as well as between ?-haemoglobinopathies (HbE and ?-thalassaemia carriers) and normal adults. Results: F cell levels varied markedly spanning 0.80-39.2 per cent with a positively skewed distribution. Thirty two per cent of these individuals had F cell levels more than the 4.5 per cent cut-off point. F cell levels in females were significantly higher than those in males (P<0.05). F cell levels in individuals having MCV <80 fl were significantly higher than those having MCV ?80 fl (P<0.05). ?-haemoglobinopathy (HbE and ?-thalassaemia carriers) had significantly higher F cell levels than normal individuals (P<0.05). Interpretation & conclusions: The present results showed that besides Swiss-type HPFH, the ?-haemoglobinopathy was expected to be involved in increased F cell levels in adult Thais. Thus, influence of ?-haemoglobinopathy must be considered in interpreting F cell levels in area endemic of this globin disorder.
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@# Objective: To investigate the effects of prostate cancer exosomes on the migration and invasion ability of stromal cells (WPMY-1), and to explore its mechanism. Methods: Exosomes in LNCaP-AI+F prostate cancer cell supernatant were isolated by ultracentrifugation and the typical structure of exosome was captured by electron microscope. The particle size distribution was analyzed by Zetaview, and Wb was used to identify the marker proteins and other proteins.After co-incubation of WPMY-1 cells and prostate cancer exosomes (40 µg/ml), laser confocal microscope was used to observe the uptake of PKH67-labeled exosomes by WPMY-1 cells; Transwell assay was used to detect the migration and invasion ability of WPMY-1 cells; qPCR was performed to detect the expression of three cancer-associated fibroblast (CAF)-related molecules (IL-8, PDGFB and MMP9) at mRNA level; and the phosphorylation of EGFR and ERK1/2 was analyzed by Wb. Results: Typical cup-shaped structure of exosomes was observed under electron microscope. The Zetaview results showed that the particle size distribution was concentrated at about 100 nm. The expression of exosome marker proteins CD63 andALIX further verified that the isolated particles were exosomes. Besides, EGFR, HER2 and SRC, which were related to the progression of prostate cancer, were also enriched in exosomes. After co-incubation, confocal microscope imaging showed a number of PKH67 labeled exosomes in recipient WPMY-1 cells. Transwell experiments showed that exosomes could significantly enhance the migration and invasion ability of WPMY-1 cells (all P<0.01). Compared with the control group, increased secretion of IL-8, PDGFB and MMP9 was observed after exosome treatment (40 µg/ml) (P<0.05 or P<0.01). Wb indicated that exosomes could promote the phosphorylation of EGFR and ERK1/2 of WPMY-1 cells (P<0.01). Conclusion: Prostate cancer cell exosomes could act on the stromal cell WPMY-1 to highly express multiple CAF-related molecules, promote the phosphorylation of EGFR and ERK1/2 and enhance the migration and invasion ability of WPMY-1 cells.
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This study aimed to describe the Macrobrachium rosenbergii hepatopancreas histomorphology. The hepatopancreas is constituted by a set of blind end tubules, divided into proximal, middle, and distal regions, with the epithelium formed by E, F, B, R, and M cells differently of other species. Measurements of the length and width of the tubules were 419.64+69.09µm and 117.42+16.99µm, respectively. The percentage of each cell type per region was: proximal region (40%B, 20%F, 6.7%M, 33.3%R), middle region (45.4%B, 18.2%F, 9.1%M, and 27.3%R) and distal region (36.4%E, 27.2%B, 18.2%F, 9.1%M, 9.1%R). Cell B that stores glycogen and lipids, is the most commonly found cell in proximal and middle regions. In the distal region, the E, responsible for the mitosis, is the most prominent. M, responsible by nutrient storage, is numerically constant among the portions differently in the Macrobrachium amazonicum. The study for the first time also suggests that in addition to digestive enzymes, the F cell produces protective mucus. The present study generated for the first time a morphometric profile of M. rosenbergii hepatopancreas, demonstrating differences from other species, and can be an important tool for new studies in nutrition, reproduction, and production with the species.(AU)
O objetivo do presente estudo foi descrever a histomorfologia do hepatopâncreas do camarão-de-água-doce Macrobrachium rosenbergii. Observou-se que ele é constituído por um conjunto de túbulos de fundo cego, sendo cada túbulo dividido em regiões proximal, média e distal, com o epitélio formado por cinco tipos de células (E, F, B, R, M), diferentemente de outras espécies. As medidas de comprimento e largura dos túbulos foram de 419,64+69,09µm e 117,42+16,99µm, respectivamente. A porcentagem de cada tipo celular por região foi: região proximal (40%B, 20%F, 6,7%M, 33,3%R), região média (45,4%B, 18,2%F, 9,1%M e 27,3%R) e região distal (36,4%E, 27,2%B, 18,2%F, 9,1%M, 9,1%R). Assim, a B, que armazena glicogênio e lipídeos, é a célula mais encontrada nas regiões proximal e média. Na região distal, a célula E, responsável pela mitose, é a mais encontrada. A célula M, responsável pelo acúmulo de nutrientes, tem um número constante de células nas porções do túbulo, diferentemente do Macrobrachium amazonicum. O estudo também sugere, pela primeira vez, que a célula F produz, além de enzimas digestivas, um muco protetor para o túbulo hepatopancreático. O presente estudo foi o primeiro a gerar um perfil morfométrico do hepatopâncreas do M. rosenbergii e demonstrou diferenças em relação a outras espécies, bem como serviu de importante ferramenta para novos estudos que abranjam a produção, a nutrição e a reprodução para a espécie.(AU)
Subject(s)
Animals , Hepatopancreas , Palaemonidae/anatomy & histology , Digestive SystemABSTRACT
Objective To investigate the effects of MTA1 knock down on migration and invasion of NPC cell 5-8F in vitro. Methods RNAi (Si-MTA1-01 and Si-MTA1-02) that can transiently silenced MTA1 was designed, synthesized and transfected into 5-8F cells by lipofectamine 2000. Control group (transfection with nonsense sequence) was also estab-lished. The efficiency of MTA1 depletion was determined by q-PCR and Western blot. Wound-healing assay ,Matrigel inva-sion assay and thesolid-phase adhesion assay were performed to investigate the effect of MTA1 knockdown on 5-8F cell me-tastasis. Results Transiently knock down of MTA1 decreased MTA1 transcription and expression in 5-8F cells compared to shRNA-con cells, showing by Real-time PCR and western blot. The invasion and migration of the cells transfected with siRNA-MTA1 were much weaker than the control group (P＜0.05). Conclusion silencing MTA 1 gene can effectively in-hibit the migration and invasion of nasopharyngeal carcinoma cell, and might be a promising target for NPC treatment.
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This study was aimed to explore differential effects of various sections of the velvet antler on promoting cell proliferation in vitro. The NRK-49F cell line from rat kidney fibroblasts was used as the cell model. The cell proliferation rates of the water extracts from the upper, middle and lower section of fresh velvet antler were measured by the MTT method. BCA method was used in the detection of protein concentration. The SDS-PAGE method was used in the analysis of difference composition of the sample protein. The results showed that soluble protein content of the upper, middle and lower section were 17.89, 16.04 and 6.89 mg·mL-1, respectively. From the top to the base, the soluble protein content of velvet antler was decreased. After 24 h treatment, when the protein concentration of the upper and middle section samples of the velvet antler were 800 μg·mL-1 and 600 μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 66.76% and 64.36%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 58.87%. After 48 h treatment, when the upper and middle section samples of the velvet antler were 800μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 219.56% and 215.86%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 169.20%. The velvet antler on the proliferation of cells was much better than the 10% fetal bovine serum. The figure of SDS-PAGE showed the slight difference in the protein composition of three part of the velvet antler. It was concluded that all samples had promoting effects on cell proliferation with concentration-depen-dent, and the main protein in different part of the velvet antler had minor differences.
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Objective To investigate the effect of nicotinamide mononucleotide (NMN) on insulin secretion and gene expressions of pancreatic and duodenal homeobox 1 ( PDX-1 ) and forkhead box-containing protein O-1 ( FoxO1 ),which were important transcription factors for insulin secretion.Methods Insulin secretion level in RIN-m5f cells was detected by rat insulin ELISA detection kit.The mRNA expression levels of PDX-1 and FoxO1 in RIN-m5f cells were analyzed by real-time PCR.The protein expression of PDX-1 was measured by Western blot.Results Insulin secretion levels in RIN-m5f cells treated with repaglinide ( 10 nmol/L) plus NMN ( 100 μnol/L) was significantly higher than those in the blank control,the DMSO control group,and the NMN (50μmol/L) treated group (P ＜0.05 ).The mRNA expression levels of PDX-1 in RIN-m5f cells treated with NMN ( 10,50 and 100 μmol/L) for 36 h were significantly higher than those in the control group (P ＜0.05,P ＜ 0.01,and P ＜ 0.001,respectively).There was marked differences in the mRNA expression levels of PDX-1 among different concentrations of NMN (P ＜0.001 ),but no significant differences in the mRNA expression level of FoxO1 ( P ＞ 0.05).No significant difference was found in the protein expression levels of PDX-1 in RIN-m5f cells treated by NMN (50,100,and 200 μmol/L) for 36 or 48 h compared with the control group (P ＞ 0.05).Conclusion NMN can stimulate insulin secretion and upregulate the mRNA expression of PDX-1 in RIN-m5f cells.
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The study was designed to examine the effect of glucose on the expression of c-myc gene in cultured RINm5F cells. After monolayer culture was established in RPMI 1640 media supplemented with 10% fetal calf serum (FCS), the cells were cultured in various concentrations of glucose and 1 or 10% FCS for another 24 hours. A mRNA was extracted from the cultured cells by a single step method, and Northern analysis was done to detect RNA band. A 0.5 kilobase single band was detected as c-myc mRNA. The expression of c-myc gene mRNA was reduced with increased concentration of glucose with 1% FCS. However, supplementation of 10% FCS abolished the effect of glucose on expression of c-myc gene. These findings suggested that glucose in conjunction with other growth promoting factors played an important role in expression of oncogene and cell growth in RINm5F cells.