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1.
Rev. bras. ciênc. avic ; 24(1): eRBCA-2021-1547, 2022. tab, graf, ilus
Article in English | VETINDEX | ID: biblio-1368398

ABSTRACT

Intramuscular fat (IMF) content is a crucial parameter for estimating meat quality. Growing evidence indicates that gene regulation plays an important role in IMF deposition. This study aimed to determine the function of Mfsd2a in chicken intramuscular preadipocytes. In the present study, high Mfsd2a mRNA levels were observed in the liver and adipose tissues of broilers. Subsequently, we synthesized small interfering RNAs to silence the expression of Mfsd2a in chicken intramuscular preadipocytes. The following results suggested that CDK2, PCNA, CCND1, CCND2 and MKI67 were inhibited, with CCK-8 and EdU assays revealing that cell proliferation was inhibited. Scratch test showed that cell migration ratios were declined. We also found that Mfsd2a silencing decreased the mRNA levels of PPARγ, RXRG and their target genes. The similar results were found in some key genes that contribute to lipid synthesis, including C/EBPα, C/EBPß, FABP4, FASN, ACACA and ACSL1. Finally, Oil red O staining showed that IMF accumulation was blocked after Mfsd2a silencing. In conclusion, our results implied that Mfsd2a promotes the proliferation and migration of chicken intramuscular preadipocytes, as well as the differentiation and adipogenesis through PPARγ signaling pathway, which may provide a potential target to improve chicken meat quality.(AU)


Subject(s)
Animals , Chickens , Proliferating Cell Nuclear Antigen , Adipogenesis , Symporters
2.
Rev. bras. ciênc. avic ; 24(4): eRBCA-2021-1589, 2022. tab, graf, ilus
Article in English | VETINDEX | ID: biblio-1382114

ABSTRACT

Lipid metabolism dysfunction is closely related to obesity, inflammation, diabetes, lipodystrophy, cardiovascular disease. Along with having a positive effect on energy homeostasis during fasting or prolonged exercise through mitochondrial fatty acid oxidation (FAO), more than two dozen enzymes and transport proteins are involved in the activation and transport of fatty acids into the mitochondrial, providing insights into their critical roles in metabolism. CPT1A has been reported to be expressed ubiquitously in the body and associated with dire consequences affecting fat deposition as the key rate-limiting enzyme of FAO. However, there is a dearth of data on the specific role of CPT1A on adipogenic differentiation and adipocyte lipolysis on chicken. This study assessed CPT1A's function in adipocyte differentiation andadipocyte lipolysis, and the mechanisms were investigated. We found that CPT1A knockdown (KD) promotes the differentiation of chicken preadipocytes into mature adipocytes. CPT1A KD increased PPARγ protein expression level. Expression levels of lipid synthesis-related genes were increased, and lipolysis genes were reduced. Also, CPT1A KD can encourage the formation of lipid droplets. So our results confirmed that knockdown of CPT1A induced the lipid differentiation and inhibited the ß-oxidation process to promote the formation of lipid droplets. These findings may deepen our understanding on CPT1A function, especially its regulatory role in adipocyte biology.(AU)


Subject(s)
Animals , Carnitine O-Palmitoyltransferase , Chickens/physiology , Adipocytes/classification , Lipid Metabolism , Gene Knockdown Techniques/instrumentation
3.
Clin Transl Oncol ; 23(11): 2394-2401, 2021 Nov.
Article in English | MEDLINE | ID: mdl-33876417

ABSTRACT

PURPOSE: This pilot study aimed on generating insight on alterations in circulating immune cells during the use of FOLFIRINOX and gemcitabine/nab-paclitaxel in pancreatic ductal adenocarcinoma (PDAC). PATIENTS AND METHODS: Peripheral blood mononuclear cells were isolated before and 30 days after initiation of chemotherapy from 20 patients with advanced PDAC. Regulatory T cells (FoxP3+) and immune checkpoints (PD-1 and TIM-3) were analyzed by flow cytometry and immunological changes were correlated with clinical outcome. RESULTS: Heterogeneous changes during chemotherapy were observed in circulating T-cell subpopulations with a pronounced effect on PD-1+ CD4+/CD8+ T cells. An increase in FoxP3+ or PD-1+ T cells had no significant effect on survival. An increase in TIM3+/CD8+ (but not TIM3+/CD4+) T cells was associated with a significant inferior outcome: median progression-free survival in the subgroup with an increase of TIM-3+/CD8+ T cells was 6.0 compared to 14.0 months in patients with a decrease/no change (p = 0.026); corresponding median overall survival was 13.0 and 20.0 months (p = 0.011), respectively. CONCLUSIONS: Chemotherapy with FOLFIRNOX or gemcitabine/nab-paclitaxel induces variable changes in circulating T-cell populations that may provide prognostic information in PDAC.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Carcinoma, Pancreatic Ductal/drug therapy , Immune Checkpoint Proteins/drug effects , Pancreatic Neoplasms/drug therapy , T-Lymphocytes, Regulatory/drug effects , Aged , Albumins/therapeutic use , CD4-Positive T-Lymphocytes/chemistry , CD4-Positive T-Lymphocytes/drug effects , CD8-Positive T-Lymphocytes/chemistry , CD8-Positive T-Lymphocytes/drug effects , Carcinoma, Pancreatic Ductal/immunology , Deoxycytidine/analogs & derivatives , Deoxycytidine/therapeutic use , Female , Fluorouracil/therapeutic use , Forkhead Transcription Factors , Hepatitis A Virus Cellular Receptor 2/analysis , Humans , Immune Checkpoint Proteins/analysis , Irinotecan/therapeutic use , Leucovorin/therapeutic use , Male , Middle Aged , Oxaliplatin/therapeutic use , Paclitaxel/therapeutic use , Pancreatic Neoplasms/immunology , Pilot Projects , Programmed Cell Death 1 Receptor/analysis , Programmed Cell Death 1 Receptor/drug effects , Progression-Free Survival , Prospective Studies , T-Lymphocytes, Regulatory/chemistry , Gemcitabine
4.
Article in English | LILACS-Express | VETINDEX | ID: biblio-1490881

ABSTRACT

ABSTRACT The present study was conducted to investigate the effects of dietary fiber on growth performance, fat deposition, serum lipids, fat metabolism, and mRNA (messenger RNA) expression of lipoprotein lipase (LPL) in Jilin white and Carlos geese. Sixty Jilin white and sixty Carlos geese aged six-weeks and of similar health and weight (average weight 313.11g) were selected. Geese of each breed were randomly divided into two groups (n=30), and with each group containing three replicate subgroups of 10 geese. The diet was supplemented with 8% or 11% fiber (corn straw powder). The Jilin white geese are divided into A1 (8%) and A2 (11%) groups, and Carlos geese are divided into B1 (8%) and B2 (11%) groups. The experiment lasted 35 days. The results showed that high dietary fiber can significantly (p 0.05) increase average daily feed intake (ADFI), significantly (p 0.05) reduce final weight (FW) and average daily gain (ADG) of both varieties, and increase LPL mRNA expression levels in abdominal fat, liver, sebum, and urethral glands. High dietary fiber accelerates intestinal peristalsis, affects the absorption of other nutrients, reduces the available energy value of the absorbed feed, and increases fat loss. Compared with the to Carlos geese, high dietary fiber content had a more significant effect on the live, slaughter, and sebum weights and sebum percentage of the Jilin white geese, indicating that the Carlos geese have higher requirements for dietary fiber content. High fiber content will reduce the growth performance, slaughter performance, and fat deposition of geese.

5.
Clin Transl Oncol ; 22(12): 2275-2285, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32447641

ABSTRACT

PURPOSE: Patients presenting with lymphovascular space invasion (LVSI) had an absolute decrease in survival. In our present study, the potential roles of LVSI on tumor characteristics was explored to predict the difference in the prognosis of ER and HER2 positive T1 tumors. METHODS: A total of 142 breast cancer patients diagnosed with ER+ and HER2+ tumors whose tumor size was ≤ 2 cm were included in this analysis. One hundred forty-two patients were divided into four groups, group 1 (lymph nodes+ and LVSI+), group 2 (lymph nodes+ and LVSI-), group 3 (lymph nodes- and LVSI+), group 4 (lymph nodes- and LVSI-). Univariate and multivariate Cox proportional hazard models were used to identify independent prognostic factors and calculate the HR and 95% CI. Kaplan-Meier and Cox regression models were used to test the prognostic significance. RESULTS: LVSI positivity was significantly associated with patient age, menopausal status, tumor size, lymph node status, Ki67, PR, and tumor grade. In the univariate and multivariate model, LVSI, PR, and Ki67 were significantly associated with DFS, and LVSI, lymph node status, PR, and Ki67 were significantly associated with OS. LVSI was significantly related to increased risk of DFS and OS only in the PR-negative and low-positive subgroups. It was a prognostic factor for DFS but not for OS in women with low Ki67 and was associated with DFS and OS in high-Ki67 tumors. Furthermore, patients who presented with only LVSI had a significantly worse survival rate than those with lymph node metastasis without LVSI in small tumors. CONCLUSION: The presence of LVSI was highlighted as a variable significant to survival. In further clinical practice, patients with LVSI may need more intensive treatment in certain populations.


Subject(s)
Breast Neoplasms/mortality , Breast Neoplasms/pathology , Receptor, ErbB-2 , Receptors, Estrogen , Adult , Age Factors , Aged , Breast Neoplasms/chemistry , Capillaries/pathology , Chemotherapy, Adjuvant , Disease-Free Survival , Female , Humans , Kaplan-Meier Estimate , Ki-67 Antigen , Lymphatic Metastasis , Lymphatic Vessels/pathology , Menopause , Middle Aged , Neoplasm Grading , Neoplasm Invasiveness , Prognosis , Proportional Hazards Models , Prospective Studies , Receptors, Progesterone , Tumor Burden , Young Adult
6.
Clin Transl Oncol ; 22(1): 91-102, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31006089

ABSTRACT

BACKGROUND: Breast ultrasound and mammography were used in the detection of residual tumor after neoadjuvant chemotherapy. The aim of this study was to evaluate the correlation between clinical and pathological responses with breast density and IHC marker conversion to understand how this information might be used in the future to direct treatment. METHODS: We included 119 patients who underwent CNB and followed NACT. The breast density assessment was based on the mammography examination performed at the time of diagnosis. We evaluated the clinical and pathological responses to NACT by the UICC and Miller-Payne grading systems, respectively. RESULTS: Of 119 patients who met the inclusion criteria, patients with high pre-treatment IHC markers levels showed higher expression of IHC markers regardless of the post-treatment IHC marker level at baseline. However, breast and node tumor sizes before and after NACT were negatively correlated with hormone receptor conversion and positively correlated with Ki-67 conversion (P < 0.05). Patients with low BD were more likely to have a cCR, pCR, TNBC, and postmenopausal status than those with a high BD (P < 0.05). BD was significantly associated with PR and Ki67 conversion but not ER conversion. CONCLUSION: Our prospective observational study demonstrated that IHC marker conversion could be used to identify lesion size changes and BD. We also found that a high BD was linked to clinical and pathological responses, molecular subtype, and menopausal status. In the future, additional studies are required to validate the predictive value identified by this research.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/metabolism , Breast Neoplasms/pathology , Chemotherapy, Adjuvant/methods , Lymph Nodes/pathology , Molecular Imaging/methods , Neoadjuvant Therapy/methods , Adult , Aged , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Female , Follow-Up Studies , Humans , Immunohistochemistry , Lymph Nodes/drug effects , Middle Aged , Prognosis , Prospective Studies , Receptor, ErbB-2/metabolism , Receptors, Estrogen/metabolism , Receptors, Progesterone/metabolism
7.
Genet Mol Res ; 16(1)2017 Mar 16.
Article in English | MEDLINE | ID: mdl-28362980

ABSTRACT

Ankylosing spondylitis (AS), a progressive disease of the spine, manifests as peripheral arthritis with tendon and ligament inflammation that restricts activity. AS is a rheumatoid autoimmune disease although the rheumatoid factor is absent in patients with AS. It is characterized by inflammatory changes such as elevated levels of serum inflammatory factors. The roles of Th1 and Th2 cytokines in autoimmune diseases are well known. However, the roles of these cytokines in the diagnosis and prognosis of AS is poorly understood. The aim of this study was to investigate the roles of Th1/Th2 cytokines in the diagnosis and prognosis of AS. The BASDAI activity, BASFI functional index, BASMI measurement score, and the levels of CRP and ESR were measured during the treatment of patients with active AS. The levels of IFN-γ and TNF-α (Th1 cytokines) and IL-4 and IL-10 (Th2 cytokines) were quantified. The levels of IL-4 and IL-10 were significantly low in the serum of patients with active AS, who also had high IFN-γ and TNF-α levels compared to those in the control individuals (P < 0.05). After treatment, the levels of IL-4 and IL-10 increased while those of IFN-γ and TNF-α decreased compared to those in individuals with active AS (P < 0.05). The disease activity index correlated positively with levels of IFN-γ and TNF-α and negatively with levels of IL-4 and IL-10, but not with that of CRP or ESR. Changes in the levels of Th1/2 cytokines in patients with AS may reflect disease activity and prognosis.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Cytokines/blood , Spondylitis, Ankylosing/diagnosis , Spondylitis, Ankylosing/drug therapy , Th1 Cells/immunology , Th2 Cells/immunology , Adolescent , Adult , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Female , Humans , Interferon-gamma/blood , Interleukin-10/blood , Interleukin-4/blood , Male , Prognosis , Severity of Illness Index , Spondylitis, Ankylosing/blood , Spondylitis, Ankylosing/immunology , Tumor Necrosis Factor-alpha/blood , Young Adult
8.
Genet Mol Res ; 16(1)2017 Mar 22.
Article in English | MEDLINE | ID: mdl-28340261

ABSTRACT

In order to evaluate the milk yield, milk quality, and health of dairy cows fed a high-concentrate (HC) diet, eight lactating Holstein dairy cattle were randomly assigned to HC or low-concentrate (LC) diet groups and fed for 50 days, and the auto-control studying before and after treatment with the two diets was used. During the experiment, plasma and milk samples were collected and measured. With regard to milk component, HC feeding led to higher milk production (P < 0.05), but significantly lower milk protein percentage (P < 0.05), milk protein yield (P < 0.05), and milk fat percentage (P < 0.05) throughout the five periods than LC feeding. Milk somatic cell count and N-acetyl-D-glucosaminidase activity (P < 0.01) were higher than those observed under LC feeding. mRNA expression levels of interleukin-8 (IL-8), C-C motif chemokine ligand (CCL5), and lactalbumin alpha (α-LA) were investigated by qPCR and found to be significantly lower (P < 0.01) in cattle fed the HC diet. The amino acid content was analyzed by high performance liquid chromatography (HPLC), and the content of Asp (P < 0.01), Gln (P < 0.01), Ala (P < 0.05), Leu (P < 0.05), Lys (P < 0.05), and Ile (P < 0.01) was significantly lower in the HC group, whereas the content of Arg (P < 0.05) and Phe (P < 0.01) was significantly higher. These results suggest that the HC diet might have an important influence on mammary health. The amino acid content was lower, suggesting that depletion of amino acids, resulting in depleted milk protein, affects milk quality.


Subject(s)
Animal Feed/analysis , Diet/veterinary , Mammary Glands, Animal/physiology , Milk Proteins/administration & dosage , Milk/standards , Actins/metabolism , Animals , Cattle , Chemokine CCL5/metabolism , Female , Interleukin-8/metabolism , Lactalbumin/metabolism , Lactation/physiology , Mammary Glands, Animal/drug effects , Mammary Glands, Animal/metabolism , Milk/metabolism , Random Allocation , Rumen/metabolism
9.
Genet Mol Res ; 16(1)2017 Mar 15.
Article in English | MEDLINE | ID: mdl-28301671

ABSTRACT

The efficacy of bone marrow mesenchymal stem cell (BMSC) on liver fibrosis in animal has been proven, but a few studies have been made in human body and few such researches in China. This study was designed to investigate the effect of BMSC treatment on hepatic fibrosis induced by hepatolenticular degeneration and the influence on serological indicators. Sixty patients with liver fibrosis induced by hepatolenticular degeneration were randomly divided into two groups, a penicillamine group and a BMSCs plus penicillamine group, with 30 patients in each. The therapeutic effects on hepatic fibrosis, liver function, and serological indicators were recorded before and after the treatment, and the data were compared. After treatment, serum levels of HA, PCIII, LN, CIV, TIMP-1, and MMP-1 were reduced in both groups (P < 0.05). However, cytokine levels in the BMSCs plus penicillamine group were significantly lower than those in the penicillamine group (P < 0.05). Combination therapy with BMSCs and penicillamine had a significant positive effect on liver fibrosis induced by hepatolenticular degeneration.


Subject(s)
Hepatolenticular Degeneration/therapy , Liver Cirrhosis/therapy , Mesenchymal Stem Cell Transplantation , Adult , Biomarkers/blood , Case-Control Studies , Cells, Cultured , Chelating Agents/administration & dosage , Combined Modality Therapy , Female , Hepatolenticular Degeneration/blood , Hepatolenticular Degeneration/complications , Humans , Liver/metabolism , Liver/pathology , Liver Cirrhosis/blood , Liver Cirrhosis/etiology , Male , Mesenchymal Stem Cells/physiology , Penicillamine/administration & dosage , Transforming Growth Factor beta1/blood , Treatment Outcome , Tumor Necrosis Factor-alpha/blood , Young Adult
10.
Genet Mol Res ; 16(1)2017 Jan 23.
Article in English | MEDLINE | ID: mdl-28128416

ABSTRACT

Inter-simple sequence repeats (ISSRs) were used to analyze the genetic diversity of 21 accessions obtained from four provinces in China, Shandong, Henan, Hebei, and Sichuan. A total of 272 scored bands were generated using the eight primers previously screened across 21 accessions, of which 267 were polymorphic (98.16%). Genetic similarity coefficients varied from 0.4816 to 0.9118, with an average of 0.6337. The UPGMA dendrogram grouped 21 accessions into two main clusters. Cluster A comprised four Lonicera macranthoides Hand. Mazz. accessions, of which J10 was found to be from Sichuan, and J17, J18, and J19 were found to be from Shandong. Cluster B comprised 17 Lonicera japonica Thumb. accessions, divided into the wild accession J16 and the other 16 cultivars. The results of the principal component analysis were comparable to the cluster analysis. Therefore, the ISSR markers could be effectively used to distinguish interspecific and intraspecific variations, which may facilitate identification of Lonicera japonica cultivars for planting, medicinal use, and germplasm conservation.


Subject(s)
Genetic Markers , Genetic Variation , Lonicera/genetics , Microsatellite Repeats , Cluster Analysis
11.
Parasite Immunol ; 39(1)2017 Jan.
Article in English | MEDLINE | ID: mdl-27801984

ABSTRACT

The objective of this study was to evaluate differential expression of innate and adaptive immune genes, including immunoglobulin, immune cell receptor, cytokine, inflammatory protein, toll-like receptors (TLR) and recombination-activating gene (RAG) in skin from channel catfish, Ictalurus punctatus after immunization with live theronts of Ichthyophthirius multifiliis (Ich) by intraperitoneal injection. The immunized catfish showed significantly higher survival rate (95%) than those of mock-immunized control fish (0% survival) after the theront challenge. The gene expression of innate immune system, such as cytokines (IL-1ß type a, IL-1ß type b, IFN-γ, TGF1-ß and TNF-α) and inflammatory proteins (NF-kB and iNOS 2), showed significant upregulation at day 1 (D1) post-immunization. Expression of TLR genes exhibited a rapid increase from hour 4 (h4) to D10 post-immunization. Genes of the adaptive response, such as the cell receptor MHC I, CD8+ , CD4+ and TCR-α, showed upregulation at D1, D6 and D10. The TCR-ß expression increased rapidly at h4 and remained upregulated until D10. Immunoglobulin IgM upregulation was detected from h4 until D2 while IgD expression was increased from D1 until D10. Rapid upregulation of innate and adaptive immune genes in skin of catfish following live theront vaccination was demonstrated in this study ultimately resulting in significant protection against Ich infection.


Subject(s)
Ciliophora Infections/veterinary , Fish Diseases/immunology , Hymenostomatida/immunology , Ictaluridae/immunology , Ictaluridae/parasitology , Skin/immunology , Animals , Antibodies, Protozoan/immunology , Ciliophora Infections/immunology , Fish Diseases/parasitology , Immunization/veterinary , Immunoglobulin M , NF-kappa B , Tumor Necrosis Factor-alpha
12.
Genet Mol Res ; 15(4)2016 Nov 03.
Article in English | MEDLINE | ID: mdl-27819716

ABSTRACT

Classic Kaposi sarcoma is a type of vascular proliferative inflammatory disease. Previous studies have reported significant associations between microRNAs expression and the development of classic Kaposi sarcoma. Here, we conducted a case-control study to investigate the association between miR-146a and miR-149 genetic polymorphisms and risk of classic Kaposi sarcoma in a Chinese population. Both classic Kaposi sarcoma patients and healthy controls were recruited between December 2013 and October 2015. Genotyping of miR-146a and miR-149 was performed by polymerase chain reaction-coupled with restriction fragment length polymorphism. Results showed that the GG genotype of miR-146a was associated with increased risk to classic Kaposi sarcoma (OR = 6.00, 95%CI = 1.19-30.12), as compared with the CC genotype. In the recessive model, we found that the GG genotype carried a 4.55-fold increased risk to classic Kaposi sarcoma as compared with the CC + CG genotype (OR = 2.06, 95%CI = 1.04-20.29). In conclusion, our study demonstrated that miR-146a, but not miR-149 polymorphism, is associated with risk to classic Kaposi sarcoma in the Chinese population.


Subject(s)
Genetic Association Studies , Genetic Predisposition to Disease , MicroRNAs/genetics , Polymorphism, Single Nucleotide/genetics , Sarcoma, Kaposi/genetics , Case-Control Studies , Demography , Female , Humans , Male , MicroRNAs/metabolism , Middle Aged , Risk Factors
13.
Genet Mol Res ; 15(4)2016 Oct 05.
Article in English | MEDLINE | ID: mdl-27808361

ABSTRACT

The pearl oyster Pinctada fucata is a commercially important marine shellfish. As a result, genetic improvement and selective-breeding program have been conducted for this species. Polymorphic microsatellites are effective molecular markers to investigate molecular marker-assisted selection and genetic variance. In this study, microsatellite DNAs were screened and characterized based on the partial genome sequence of P. fucata. We identified 111 microsatellite DNA motifs through mining the published draft genome sequence of P. fucata. Forty-two loci were screened with 8 P. fucata individuals, and 15 were found to be polymorphic and were therefore further evaluated using 40 wild individuals from the Daya Bay, Shenzhen City, Guangdong Province, China. The number of alleles per locus ranged from 3 to 8, with an average of 5.2667 for the 15 polymorphic loci. Observed and expected heterozygosities ranged from 0.1154 to 0.6216 (0.3321 on average) and 0.4950 to 0.8491 (0.6768 on average), respectively. Of the 15 polymorphic loci, 12 loci deviated from Hardy-Weinberg equilibrium after Bonferroni correction (P < 0.0033). Polymorphism information content ranged from 0.44 to 0.83 with a mean value of 0.63. The results suggest that the markers isolated in this study can be used for research on molecular marker-assisted selection and genetic variance of P. fucata.


Subject(s)
Genetic Loci , Microsatellite Repeats/genetics , Pinctada/genetics , Polymorphism, Genetic , Animals , Genetic Testing
14.
Genet Mol Res ; 15(3)2016 Sep 02.
Article in English | MEDLINE | ID: mdl-27706682

ABSTRACT

The development of age-related cardiovascular disease is associated with the senescence of vascular cells. This study aimed to investigate the effect of ginsenoside Rg1 on vascular smooth muscle cell (VSMC) senescence. Primary VSMCs were cultured and divided into control, D-galactose (D-gal), Rg1-L, and Rg1-H groups, which were cultured without and with D-gal, and with low- and high-concentrations of Rg1, respectively. D-gal-induced cellular senescence was identified by b-galactosidase staining, and ultrastructural changes within the cells were observed. The expression of p16, p21, and p53 in the four groups of VSMCs was determined by western blotting, and the cell cycle was investigated by flow cytometry. Compared with the control group, there was an obvious change in the ultrastructure of VSMCs in the D-gal group, and the proportion of b-galactosidase-positive cells was significantly increased (P < 0.05). In addition, p16, p21, and p53 expression was significantly increased (P < 0.05) and the cell cycle was arrested in the G0/G1 phase. Compared with the D-gal group, the percentage of positive cells was significantly reduced (P < 0.05) in the Rg1 groups, the expression of p16, p21, and p53 was significantly reduced (P < 0.05), and the number of cells in the G0/G1 phase decreased (P < 0.05). Ginsenoside Rg1 can inhibit VSMC senescence, and the mechanisms may be related to its partial inhibition of the p16INK4a/Rb and p53-p21Cip1/Waf1 signaling pathways during the cell cycle.


Subject(s)
Cellular Senescence/drug effects , G1 Phase Cell Cycle Checkpoints/drug effects , Ginsenosides/pharmacology , Myocytes, Smooth Muscle/drug effects , Animals , Cell Adhesion/drug effects , Cell Cycle/drug effects , Cell Cycle/genetics , Cell Proliferation/drug effects , Cyclin-Dependent Kinase Inhibitor p16/genetics , Cyclin-Dependent Kinase Inhibitor p16/metabolism , Cyclin-Dependent Kinase Inhibitor p21/genetics , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Drugs, Chinese Herbal/chemistry , Galactose/pharmacology , Gene Expression Regulation , Ginsenosides/isolation & purification , Male , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , Primary Cell Culture , Rats , Rats, Sprague-Dawley , Signal Transduction , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , beta-Galactosidase/genetics , beta-Galactosidase/metabolism
15.
Genet Mol Res ; 15(3)2016 Sep 23.
Article in English | MEDLINE | ID: mdl-27706781

ABSTRACT

Copper-transporting P-type adenosine triphosphatase (ATP7B) has been identified as the pathogenic gene in hepatolenticular degeneration, or Wilson's disease (WD). The aim of this study was to explore the correlation between genetic mutations and the clinical profile of WD, and to discuss the value of mutation examination in its diagnosis for providing a scientific basis for the development of a method to examine genetic mutations. Sixty-eight Chinese Han patients with WD and 20 controls were included in this study. The ATP7B gene in DNA extracted from patient blood samples was amplified by PCR and sequenced. These sequences were compared against corresponding gene sequences obtained from healthy controls to statistically analyze the genetic mutations. Five of the nineteen mutations in ATP7B were newly detected mutations; moreover, 8 of these mutations were polymorphic (2 were newly identified). The Arg778Leu and Pro992Leu mutations in exons 8 and 13 were detected at the highest mutation frequencies of 25.74 and 16.91%, respectively. The frequencies of all other mutations were below 5%. However, the clinical manifestations of WD did not differ significantly in patients with the Arg778Leu and Pro992Leu mutations. Therefore, these mutations were considered as hotspot mutations in Chinese WD patients. However, we observed no significant correlation between these genetic types and the clinical symptoms of WD. The correlation between the mutation genotype and disease phenotype remains to be elucidated. In conclusion, the highly sensitive and specific direct DNA sequencing method can be used to screen for the causative genes of WD.


Subject(s)
Adenosine Triphosphatases/genetics , Cation Transport Proteins/genetics , Copper/metabolism , Hepatolenticular Degeneration/diagnosis , Hepatolenticular Degeneration/genetics , Mutation/genetics , Sequence Analysis, DNA/methods , Base Sequence , Copper-Transporting ATPases , Exons/genetics , Female , Genetic Association Studies , Humans , Male , Polymorphism, Genetic
16.
Genet Mol Res ; 15(3)2016 Jul 29.
Article in English | MEDLINE | ID: mdl-27525896

ABSTRACT

Congenital cataract is a common cause of blindness in children; however, its pathogenesis remains unclear. Genetic factors have been shown to play an important role in the pathogenesis of congenital cataract. The current genetic models of congenital cataract include autosomal dominant, autosomal recessive, and sex-linked inheritance. Sex-linked congenital cataract could be inherited through the X or Y chromosome. Congenital cataract is a symptom associated with several X-linked disorders, including Nance-Horan syndrome, Lowe syndrome, Conradi-Hünermann-Happle syndrome, oculo-facio-cardio-dental syndrome, and Alport syndrome. On the other hand, the mechanism and characteristics of Y-linked congenital cataract remains to be identified. Despite its rarity, sex-linked congenital cataract has been known to seriously affect the quality of life of patients. In this review, we present our current understanding of the genes and loci associated with sex-linked congenital cataract. This could help identify novel approaches for the prevention, early diagnosis, and comprehensive disease treatment.


Subject(s)
Cataract/genetics , Genetic Diseases, X-Linked/genetics , DNA Mutational Analysis , Genes, X-Linked , Genetic Association Studies , Genetic Loci , Genetic Predisposition to Disease , Humans , Mutation
17.
Genet Mol Res ; 15(2)2016 Jun 21.
Article in English | MEDLINE | ID: mdl-27420955

ABSTRACT

Male ICR mice were orally administered samarium nitrate [Sm(NO3)3] to investigate its effects on sperm concentration and sperm quality. After acute exposure to ≥2880.00 mg/kg Sm(NO3)3 via intragastric gavage, sperm motility and acrosome integrity were decreased, and the sperm malformation percentage was increased (P < 0.05). After subchronic exposure to ≥500.00 mg/L Sm(NO3)3 administered via drinking water for 90 days, relative gonad weight, sperm concentration, and sperm quality significantly decreased (P < 0.05). Sperm malformation also increased after subchronic exposure to Sm, which was found to be the most sensitive index. Sperm head malformation accounted for the largest proportion of all types of sperm malformations evaluated. Of the six different subtypes of head malformation, irregular shape accounted for the largest proportion.


Subject(s)
Acrosome/drug effects , Samarium/toxicity , Sperm Motility/drug effects , Acrosome/pathology , Animals , Male , Mice , Mice, Inbred ICR , Samarium/administration & dosage , Sperm Count
18.
Genet Mol Res ; 15(2)2016 Jul 14.
Article in English | MEDLINE | ID: mdl-27421010

ABSTRACT

Studying thermotolerance is important for the prevention of thermostress in chickens. This study aimed to analyze the effect of mutations in the heat shock protein 70 (HSP70) gene on chicken thermotolerance. The C.-69A>G SNP in the 5'-flanking region of the HSP70 gene was genotyped in Lingshan and White Recessive Rock (WRR) chickens. Association of this SNP with thermotolerance traits revealed it to be significantly associated with CD4+/CD8+, and potentially associated with heterophil-to-lymphocyte ratio in WRR chickens exposed to thermoneutral temperature (15°C). Online prediction detected a putative myeloid zinc finger protein 1 binding factor in the C.-69A>G mutation. Under acute thermostress, mRNA levels of HSP70 in individuals with different C.-69A>G genotypes varied in the heart, leg muscle, and liver tissues. The HSP70 protein was expressed at higher levels in individuals with the GG genotype than in those with the AA genotype. In heart and liver, protein expression of HSP70 in individuals with the GG genotype was significantly higher than in those with the AA genotype. In leg muscle, protein expression was higher in birds with the GG genotype than in those with the AA and AG genotypes. Luciferase activity of the GG genotype was significantly higher than that of the AA genotype, suggesting that the C.-69A>G SNP regulates HSP70 gene expression. These results indicate that the C.-69A>G SNP in the 5'-flanking region of the HSP70 gene might affect chicken thermotolerance and that the GG genotype might be advantageous for the prevention of thermostress.


Subject(s)
Chickens/physiology , HSP70 Heat-Shock Proteins/genetics , Thermotolerance/genetics , 5' Flanking Region , Animals , Chickens/genetics , Female , Genotype , Mutation , Polymorphism, Single Nucleotide , RNA, Messenger/metabolism
19.
Braz J Med Biol Res ; 49(8)2016 Jul 11.
Article in English | MEDLINE | ID: mdl-27409336

ABSTRACT

Fluoride, which is often added to toothpaste or mouthwash in order to protect teeth from decay, may be a novel therapeutic approach for acceleration of periodontal regeneration. Therefore, we investigated the effects of fluoride on proliferation and mineralization in human periodontal ligament cells in vitro. The periodontal ligament cells were stimulated with various concentrations of NaF added into osteogenic inductive medium. Immunohistochemistry of cell identification, cell proliferation, alkaline phosphatase (ALP) activity assay, Alizarin red S staining and quantitative real-time-polymerase chain reaction (RT-PCR) were performed. Moderate concentrations of NaF (50-500 µmol/L) had pro-proliferation effects, while 500 µmol/L had the best effects. ALP activity and calcium content were significantly enhanced by 10 µmol/L NaF with osteogenic inductive medium. Quantitative RT-PCR data varied in genes as a result of different NaF concentrations and treatment periods. We conclude that moderate concentrations of NaF can stimulate proliferation and mineralization in periodontal ligament cells. These in vitro findings may provide a novel therapeutic approach for acceleration of periodontal regeneration by addition of suitable concentrations of NaF into the medication for periodontitis treatment, i.e., into periodontal packs and tissue patches.


Subject(s)
Cell Proliferation/drug effects , Periodontal Ligament/drug effects , Sodium Fluoride/pharmacology , Adolescent , Adult , Alkaline Phosphatase/drug effects , Alkaline Phosphatase/metabolism , Calcium/metabolism , Cells, Cultured/drug effects , Child , Humans , Periodontal Ligament/cytology , Real-Time Polymerase Chain Reaction/methods , Young Adult
20.
Genet Mol Res ; 15(2)2016 May 09.
Article in English | MEDLINE | ID: mdl-27173355

ABSTRACT

Liver cancer is a common malignant tumor associated with a short-survival period and high-mortality rate, and its prevalence in China is particularly high. This study aimed to investigate the effect of overexpressing the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene on liver cancer cell apoptosis and provide new insight into the treatment of this disease. The experimental design included four treatment groups, consisting of HHCC and H22 cells transfected with PTEN recombinant plasmids (HHCC+PTEN, H22+PTEN), and those transfected with control plasmids (HHCC+NC, H22+NC). The expression of PTEN mRNA was determined by quantitative polymerase chain reaction, and protein levels were examined by western blot. Cell apoptosis was measured using flow cytometry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. PTEN mRNA expression in cells transfected with pcDNA3.1-PTEN was significantly increased compared to the control groups (P < 0.05). In addition, western blotting revealed PTEN protein expression in the treatment groups to be significantly elevated in comparison to control cells (P < 0.05). Flow cytometry showed that apoptosis rates of both HHCC+PTEN (approximately 21.9%) and H22+PTEN (approximately 41.0%) cells were significantly higher than those of the control groups (P < 0.05). Moreover, the difference in apoptosis rate between experimental and control groups was significant (P < 0.05). In this study, HHCC and H22 cells were successfully transfected with pcDNA3.1-PTEN in vitro. We conclude that overexpression of PTEN can effectively inhibit proliferation of these cells and promote their apoptosis.


Subject(s)
Carcinoma, Hepatocellular/enzymology , Liver Neoplasms/enzymology , PTEN Phosphohydrolase/biosynthesis , Animals , Apoptosis/physiology , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , China , Humans , In Situ Nick-End Labeling , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Liver Neoplasms, Experimental/enzymology , Liver Neoplasms, Experimental/genetics , Liver Neoplasms, Experimental/pathology , Mice , PTEN Phosphohydrolase/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Transfection/methods
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