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1.
Genet Mol Res ; 16(2)2017 Jun 29.
Article in English | MEDLINE | ID: mdl-28671245

ABSTRACT

Acid and pepsin-soluble collagen (ASC and PSC, respectively) were extracted from cod (Gadus macrocephaius) skin, and yields of 37.36 and 55.96% were obtained for ASC and PSC, respectively. The total yield of ASC and PSC was 93.92%, based on the lyophilized dry weight, which is higher than that obtained from other sources. Electrophoresis revealed that both ASC and PSC consisted of two different α-chains (α1 and α2), which were characterized as type I collagen. Analysis of amino acids showed that both the ASC and PSC contained imino acids (216.1 and 190.6 residues/1000 residues, respectively) and the Fourier transform infrared spectroscopy spectra of both collagens were similar with pepsin hydrolysis having no effect on their triple-helical structure. The thermal denaturation temperature of ASC and PSC, as measured by viscometry, was 26.8° and 25.6°C, respectively.


Subject(s)
Collagen/chemistry , Fish Proteins/chemistry , Amino Acids/analysis , Animals , Gadiformes , Protein Denaturation , Protein Subunits/chemistry , Skin/chemistry
2.
Braz J Med Biol Res ; 50(3): e5796, 2017 Mar 02.
Article in English | MEDLINE | ID: mdl-28273209

ABSTRACT

We aimed to investigate the influence of regulatory T cells including CD4+CD25+, CD8+CD28- and hepatitis B virus (HBV) genotype on sustained virological response and tolerance of nucleoside drugs. One hundred and thirty-seven patients were enrolled. Lamivudine was administered to 84 patients. Entecavir was administered to the other 53 patients. Before treatment, biochemical tests, HBV DNA load, HBV serum level, HBV genotype, PB CD3+, CD4+, CD8+, CD4+CD25+/CD3+, and CD8+CD28-/CD3+ frequencies were measured. Based on HBV DNA loads after 4 weeks of therapy, patients were divided into response group and suboptimal response group. The lamivudine group received treatment continuously, and then patients were categorized into non-resistance group and resistance group. Compared with the suboptimal response and resistance groups for lamivudine, CD4+CD25+/CD3+ levels were higher in the response and non-resistance groups (t=4.372, P=0.046; t=7.262, P=0.017). In the non-resistance group, CD8+CD28-/CD3+ frequency was lower than in the resistance group (t=5.527, P=0.037). Virus load and hepatitis B E antigen (HBeAg)-positive rate were significantly lower than in the response and resistance group (t=2.164, P=0.038; X2=4.239, P=0.040; t=2.015, P=0.044; X2=16.2, P=0.000). Incidence of drug resistance was high in patients with virogene type C. For the virological response to entecavir, CD8+CD28-/CD3+ level was significantly lower than that of the suboptimal response group (t=6.283, P=0.036). Response and suboptimal response groups were compared in CD3+, CD4+, CD8+, CD4+CD25+/CD3+ and virus genotype, and differences were not statistically significant (P>0.05). Baseline regulatory T cells including CD4+CD25+/CD3+ and CD8+CD28-/CD3+ frequencies have a relationship with the incidence of rapid virological response and the resistance to nucleoside drugs. Patients with HBV genotype C receiving lamivudine more often underwent drug resistance. Antiviral efficacy and the resistance to lamivudine were closely correlated with baseline factors; the same cannot be found for entecavir.


Subject(s)
Antiviral Agents/therapeutic use , Guanine/analogs & derivatives , Hepatitis B virus/immunology , Hepatitis B, Chronic/drug therapy , Lamivudine/therapeutic use , Nucleosides/therapeutic use , T-Lymphocytes, Regulatory , Adult , Aged , Drug Resistance , Female , Genotype , Guanine/therapeutic use , Hepatitis B virus/drug effects , Hepatitis B, Chronic/virology , Humans , Male , Middle Aged , Sustained Virologic Response , T-Lymphocytes, Regulatory/immunology , Time Factors
3.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;50(3): e5796, 2017. tab
Article in English | LILACS | ID: biblio-839267

ABSTRACT

We aimed to investigate the influence of regulatory T cells including CD4+CD25+, CD8+CD28- and hepatitis B virus (HBV) genotype on sustained virological response and tolerance of nucleoside drugs. One hundred and thirty-seven patients were enrolled. Lamivudine was administered to 84 patients. Entecavir was administered to the other 53 patients. Before treatment, biochemical tests, HBV DNA load, HBV serum level, HBV genotype, PB CD3+, CD4+, CD8+, CD4+CD25+/CD3+, and CD8+CD28-/CD3+ frequencies were measured. Based on HBV DNA loads after 4 weeks of therapy, patients were divided into response group and suboptimal response group. The lamivudine group received treatment continuously, and then patients were categorized into non-resistance group and resistance group. Compared with the suboptimal response and resistance groups for lamivudine, CD4+CD25+/CD3+ levels were higher in the response and non-resistance groups (t=4.372, P=0.046; t=7.262, P=0.017). In the non-resistance group, CD8+CD28-/CD3+ frequency was lower than in the resistance group (t=5.527, P=0.037). Virus load and hepatitis B E antigen (HBeAg)-positive rate were significantly lower than in the response and resistance group (t=2.164, P=0.038; X2=4.239, P=0.040; t=2.015, P=0.044; X2=16.2, P=0.000). Incidence of drug resistance was high in patients with virogene type C. For the virological response to entecavir, CD8+CD28-/CD3+ level was significantly lower than that of the suboptimal response group (t=6.283, P=0.036). Response and suboptimal response groups were compared in CD3+, CD4+, CD8+, CD4+CD25+/CD3+ and virus genotype, and differences were not statistically significant (P>0.05). Baseline regulatory T cells including CD4+CD25+/CD3+ and CD8+CD28-/CD3+ frequencies have a relationship with the incidence of rapid virological response and the resistance to nucleoside drugs. Patients with HBV genotype C receiving lamivudine more often underwent drug resistance. Antiviral efficacy and the resistance to lamivudine were closely correlated with baseline factors; the same cannot be found for entecavir.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Antiviral Agents/therapeutic use , Guanine/analogs & derivatives , Hepatitis B virus/immunology , Hepatitis B, Chronic/drug therapy , Lamivudine/therapeutic use , Nucleosides/therapeutic use , T-Lymphocytes, Regulatory , Drug Resistance , Genotype , Guanine/therapeutic use , Hepatitis B virus/drug effects , Hepatitis B, Chronic/virology , Sustained Virologic Response , T-Lymphocytes, Regulatory/immunology , Time Factors
4.
Genet Mol Res ; 14(4): 13364-9, 2015 Oct 27.
Article in English | MEDLINE | ID: mdl-26535650

ABSTRACT

We conducted a study to investigate the role of three IL-17 gene single nucleotide polymorphisms (SNP) (rs2275913G>A, rs3748067C>T, and rs763780 T>C) in the development of gastric cancer. A total of 252 patients with gastric cancer and 252 control subjects were collected between May 2012 and May 2014. The SNP genotyping of IL-17A rs2275913G>A and rs3748067C>T and IL-17F rs763780 T>C was performed using the Sequenom MassARRAY platform (Sequenom, San Diego, CA, USA) according to the manufacturer instructions. By conditional regression analysis, individuals carrying the AA and the GA+AA genotypes of rs2275913G>A were correlated with an elevated risk of gastric cancer when compared with those carrying the GG genotype, and the adjusted ORs (95%CIs) were 2.05 (1.13-3.76) for the AA genotype and 1.45 (1.03-2.08) for the GA+AA genotype. In conclusion, our results suggest that the IL-17A rs3748067C>T and IL-17F rs763780 T>C polymorphisms play an important role in the risk of gastric cancer in a Chinese population.


Subject(s)
Genetic Predisposition to Disease , Interleukin-17/genetics , Polymorphism, Single Nucleotide , Stomach Neoplasms/genetics , Aged , Alleles , Case-Control Studies , Female , Gene Frequency , Genotype , Humans , Male , Middle Aged , Risk Factors
5.
Genet Mol Res ; 14(1): 1293-300, 2015 Feb 13.
Article in English | MEDLINE | ID: mdl-25730068

ABSTRACT

Numerous studies have evaluated the association between estrogen receptor alpha (ESR1) gene PvuII polymorphism and fracture risk in postmenopausal women. However, the results have been inconsistent. We performed a meta-analysis to examine the association between the ESR1 gene PvuII polymorphism and fracture risk in postmenopausal women. Studies published from PubMed, Google Scholar, and China National Knowledge Infrastructure data were retrieved. Pooled odds ratios with 95% confidence intervals were calculated using fixed- or random-effects models. A total of 6 case-control studies containing 592 patients and 705 controls were included in this meta-analysis. We found no association between the PvuII polymorphism in the ESR1 gene and fracture in postmenopausal women. Taking into account the effect of ethnicity, further stratified analyses were performed. In the subgroup analysis, no significant association was found in Caucasians and in Asians. No publication bias was found in the present study (all P > 0.05). In conclusion, the ESR1 gene PvuII polymorphism may not be associated with fracture risk in postmenopausal women. Additional larger studies are needed to confirm this conclusion.


Subject(s)
Deoxyribonucleases, Type II Site-Specific/genetics , Estrogen Receptor alpha/genetics , Fractures, Bone/genetics , Genetic Predisposition to Disease/genetics , Osteoporotic Fractures/genetics , Polymorphism, Genetic , Aged , Asian People , Case-Control Studies , Female , Fractures, Bone/ethnology , Genotype , Humans , Introns , Odds Ratio , Osteoporosis, Postmenopausal/ethnology , Osteoporosis, Postmenopausal/genetics , Osteoporotic Fractures/ethnology , Postmenopause , Risk Factors , White People
6.
Genet Mol Res ; 13(2): 4083-8, 2014 May 30.
Article in English | MEDLINE | ID: mdl-24938700

ABSTRACT

Fas/FasL protein expression of bone marrow hematopoietic cells was investigated in severe aplastic anemia (SAA) patients. Fas expression was evaluated in CD34(+), GlycoA(+), CD33(+), and CD14(+) cells labeled with monoclonal antibodies in newly diagnosed and remission SAA patients along with normal controls. FasL expression was evaluated in CD8(+) cells in the same manner. In CD34(+) cells, Fas expression was significantly higher in the newly diagnosed SAA group (46.59 ± 27.60%) than the remission (6.12 ± 3.35%; P < 0.01) and control (8.89 ± 7.28%; P < 0.01) groups. In CD14(+), CD33(+), and GlycoA(+) cells, Fas levels were significantly lower in the newly diagnosed SAA group (29.29 ± 9.23, 46.88 ± 14.30, and 15.15 ± 9.26%, respectively) than in the remission (47.23 ± 31.56, 67.22 ± 34.68, and 43.56 ± 26.85%, respectively; P < 0.05) and normal control (51.25 ± 38.36, 72.06 ± 39.88, 50.38 ± 39.88%, respectively; P < 0.05) groups. FasL expression of CD8(+) cells was significantly higher in the newly diagnosed SAA group (89.53 ± 45.68%) than the remission (56.39 ± 27.94%; P < 0.01) and control (48.63 ± 27.38%; P <0.01) groups. No significant differences were observed between the remission and control groups. FasL expression in CD8(+) T cells was significantly higher in newly diagnosed patients, and CD34(+), CD33(+), CD14(+), and GlycoA(+) cells all showed Fas antigen expression. The Fas/FasL pathway might play an important role in excessive hematopoietic cell apoptosis in SAA bone marrow. Furthermore, CD34(+) cells are likely the main targets of SAA immune injury.


Subject(s)
Anemia, Aplastic/genetics , Apoptosis Regulatory Proteins/genetics , Bone Marrow Cells/metabolism , Fas Ligand Protein/genetics , Hematopoietic Stem Cells/metabolism , Adolescent , Adult , Anemia, Aplastic/immunology , Anemia, Aplastic/pathology , Antigens, CD34/biosynthesis , Antigens, CD34/genetics , Apoptosis/immunology , Apoptosis Regulatory Proteins/biosynthesis , Bone Marrow Cells/immunology , CD8 Antigens/biosynthesis , CD8 Antigens/genetics , Fas Ligand Protein/biosynthesis , Female , Gene Expression Regulation, Neoplastic , Hematopoietic Stem Cells/immunology , Humans , Leukocyte Common Antigens/biosynthesis , Leukocyte Common Antigens/genetics , Male
7.
Clin Transl Oncol ; 16(3): 307-14, 2014 Mar.
Article in English | MEDLINE | ID: mdl-23873251

ABSTRACT

PURPOSE: In the present study, we intend to detect the expression of Forkhead box transcription (FOXM1) in gastric cancer tissues and cell lines, and analyze the correlation between FOXM1 expression and clinic-pathological features as well as their association with clinic outcomes in patients with resectable gastric cancers. METHODS: We examined the expression of FOXM1 in 103 cancer tissues from patients who underwent gastrectomy during Jan 2007 to Nov 2007 and 68 randomly selected para-cancer tissues by immunohistochemistry. The expression of FOXM1 protein in the benign and malignant human gastric cell lines was simultaneously detected using Western blot analysis. Data on clinic-pathological features and relevant prognostic factors in these patients were then analyzed. RESULTS: FOXM1 expression was absolutely higher in gastric cancer than para-cancer tissues (P < 0.001) and normal gastric epithelium cell lines (P = 0.022). No significant association was found between FOXM1 expression and any clinic-pathological parameters (P > 0.1). FOXM1 amplification was showed to be independently associated with prognosis in gastric cancer patients (P = 0.001), and its affection is more significant in patients with tumor size larger than 5 cm (P = 0.004), pT3-4 (P = 0.003) or pIII-IV (P = 0.001) as a result of stage-stratified analysis. CONCLUSIONS: Overexpressed FOXM1 is a potential diagnostic and poor prognostic biomarker in postoperational gastric cancer patients.


Subject(s)
Adenocarcinoma/metabolism , Biomarkers, Tumor/analysis , Forkhead Transcription Factors/analysis , Stomach Neoplasms/metabolism , Adenocarcinoma/mortality , Adenocarcinoma/pathology , Blotting, Western , Female , Forkhead Box Protein M1 , Gastrectomy , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Male , Middle Aged , Prognosis , Proportional Hazards Models , Stomach Neoplasms/mortality , Stomach Neoplasms/pathology
8.
Braz J Med Biol Res ; 32(7): 885-9, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10454748

ABSTRACT

The treatment of some mesenchymal malignancies has made significant gains over the past few decades with the development of effective systemic therapies. In contrast, the treatment of chondrosarcoma has been limited to surgical resection, with the most significant prognostic indicators being surgical margins and histologic grade. We have reported that MMP-1/TIMP-1 gene expression serves to prognosticate for tumor recurrence in this group of patients. This led to the hypothesis that collagenase activity facilitates cell egression from the cartilaginous matrix. In the current study we examine the specificity of collagenase gene expression in archival human chondrosarcoma samples using semi-quantitative PCR. Messenger RNA was affinity extracted and subject to reverse transcription. The subsequent cDNA was amplified using novel primers and quantitated by densitometry. Ratios of gene expression were constructed and compared to disease-free survival. The data demonstrate that the significance of the MMP-1/TIMP-1 ratio as a predictor of recurrence is confirmed with a larger number of patients. Neutrophil collagenase or MMP-8 was observed in only 5 of 29 samples. Collagenase-3 or MMP-13 was observed in all samples but the level did not correlate with disease-free survival. Since the collagenases have similar activity for fibrillar collagens and cleave the peptide in the same location, post-transcriptional regulatory mechanisms may account for the observed specificity. The determination of the MMP-1/TIMP-1 gene expression ratio not only serves to identify those patients at risk for recurrence but may also serve as a novel therapeutic avenue as an adjunct to surgical resection.


Subject(s)
Bone Neoplasms/enzymology , Bone Neoplasms/secondary , Chondrosarcoma/enzymology , Chondrosarcoma/secondary , Collagenases/metabolism , Gene Expression Regulation, Enzymologic , Collagenases/analysis , Collagenases/genetics , Disease-Free Survival , Humans , Prognosis , Substrate Specificity/genetics
9.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;32(7): 885-9, July 1999.
Article in English | LILACS | ID: lil-234895

ABSTRACT

The treatment of some mesenchymal malignancies has made significant gains over the past few decades with the development of effective systemic therapies. In contrast, the treatment of chondrosarcoma has been limited to surgical resection, with the most significant prognostic indicators being surgical margins and histologic grade. We have reported that MMP-1/TIMP-1 gene expression serves to prognosticate for tumor recurrence in this group of patients. This led to the hypothesis that collagenase activity facilitates cell egression from the cartilaginous matrix. In the current study we examine the specificity of collagenase gene expression in archival human chondrosarcoma samples using semi-quantitative PCR. Messenger RNA was affinity extracted and subject to reverse transcription. The subsequent cDNA was amplified using novel primers and quantitated by densitometry. Ratios of gene expression were constructed and compared to disease-free survival. The data demonstrate that the significance of the MMP-1/TIMP-1 ratio as a predictor of recurrence is confirmed with a larger number of patients. Neutrophil collagenase or MMP-8 was observed in only 5 of 29 samples. Collagenase-3 or MMP-13 was observed in all samples but the level did not correlate with disease-free survival. Since the collagenases have similar activity for fibrillar collagens and cleave the peptide in the same location, post-transcriptional regulatory mechanisms may account for the observed specificity. The determination of the MMP-1/TIMP-1 gene expression ratio not only serves to identify those patients at risk for recurrence but may also serve as a novel therapeutic avenue as an adjunct to surgical resection


Subject(s)
Humans , Bone Neoplasms/enzymology , Chondrosarcoma/enzymology , Collagenases/metabolism , Gene Expression Regulation, Enzymologic , Collagenases/analysis , Collagenases/genetics , Disease-Free Survival , DNA, Complementary/analysis , Polymerase Chain Reaction , Prognosis , Recurrence/prevention & control , RNA, Messenger/analysis , Substrate Specificity/genetics , Tissue Inhibitor of Metalloproteinase-1/analysis
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