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1.
J Clin Lab Anal ; : e22967, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31265177

RESUMO

BACKGROUND: The protein encoded by the selenoprotein S gene is considered to be an anti-inflammatory and antioxidant protein and is involved in a variety of diseases. Therefore, we want to study the distribution characteristics of this gene in Chinese diabetic population. METHODS: A total of 170 patients with DM (including 100 patients with T2DM and 70 patients with diabetic nephropathy [DN]) and 100 healthy controls (HC) were selected from Haikou People's Hospital (China) between January 2017 and July 2017. The polymorphisms of three SEPS1 genes (SNP ID: rs4965814, rs28665122, and rs34713741) were measured by massARRAY method, while the polymorphisms of SEPS1 genes (SNP ID: rs4965373) were detected by Sanger sequencing. RESULTS: Comparing three groups, the results were the following: (a) There was a significant difference in the genotype and allele distribution of rs34713741 between DN group and HC group and between T2DM group and DN group; For this gene locus, the risk of diabetic nephropathy in healthy individuals with T allele was 0.6 times higher than that in individuals with GG genotype (OR = 0.60, 95% CI: 0.46 ~ 0.77). (b) There was a significant difference in the distribution of rs4975814 genotype between DN group and HC group; for this gene locus, the risk of diabetic nephropathy in healthy individuals with T allele was 2.71 times higher than that in individuals with GG genotype (OR = 2.71, 95% CI: 1.66 ~ 4.45). CONCLUSION: We conclude that rs34713741 (GT + TT) may be a protective gene for DN and the rs4975814 (GT + TT) may be a susceptibility gene for DN.

2.
J Biol Chem ; 2018 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-30425099

RESUMO

LATERAL ORGAN BOUNDARIES DOMAIN (LBD) proteins, a family of plant-specific transcription factors harboring a conserved LOB domain, are regulators of plant organ development. Recent studies have unraveled additional pivotal roles of the LBD protein family beyond defining lateral organ boundaries, such as pollen development and nitrogen metabolism. The structural basis for the molecular network of LBD-dependent processes remains to be deciphered. Here, we solved the first structure of the homodimeric LOB domain of Ramosa2 from wheat (TtRa2LD) to 1.9 Å resolution. Our crystal structure reveals structural features shared with other zinc-finger transcriptional factors, as well as some features unique to LBD proteins. Formation of the TtRa2LD homodimer relied on hydrophobic interactions of its coiled-coil motifs. Several specific motifs/domains of the LBD protein were also involved in maintaining its overall conformation. The intricate assembly within and between the monomers determined the precise spatial configuration of the two zinc fingers that recognize palindromic DNA sequences. Biochemical, molecular modeling, and small-angle X-ray scattering (SAXS) experiments indicated that dimerization is important for cooperative DNA binding and discrimination of palindromic DNA through a molecular calipers mechanism. Along with previously published data, this study enables us to establish an atomic-scale mechanistic model for LBD proteins as transcriptional regulators in plants.

3.
Adv Exp Med Biol ; 1068: 119-133, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29943300

RESUMO

Hematological malignancies (HM) are a heterogeneous group of life-threatening hematological diseases. The heterogeneity and clonal evolution of HM subpopulations are the main obstacles for precise diagnoses, risk stratification, and even targeted therapies. Standard bulk-sample genomic examinations average total mutations from multiple subpopulations and conceal the clonal diversity that may play a significant role in HM progression. Therefore, the development of novel methods that detect intra-tumor heterogeneity is critical for the discovery of novel potential therapeutic targets. The recently developed single cell sequencing (SCS) technologies can analyse genetic polymorphisms at a single cell level. SCS requires the precise isolation of single cells and amplification of their genetic material. It allows the analysis of genomic, transcriptomic, and epigenomic information in single cancer cells. SCS may also be able to monitor minimal residual disease (MRD) of HM by sequencing circulating tumor cells (CTCs) from peripheral blood. Functional heterogeneity and clonal evolution exist in acute leukemia, multiple myeloma (MM) and chronic myeloid leukemia (CML) subpopulations and have prognostic value. In this thesis, we provide an overview of SCS technologies in HM and discuss the heterogeneous genetic variation and clonal structure among subpopulations of HM. Furthermore, we aimed to shed light on the clinical applications of SCS technologies, including the development of new targeted therapies for drug-resistant or recurrent HM.

4.
Sci Rep ; 7: 42865, 2017 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-28216645

RESUMO

3'-5' exonucleases are frequently found to be associated to polymerases or helicases domains in the same enzyme or could function as autonomous entities. Here we uncovered that Candida albicans Pif1 (CaPif1) displays a 3'-5' exonuclease activity besides its main helicase activity. These two latter activities appear to reside on the same polypeptide and the new exonuclease activity could be mapped to the helicase core domain. We clearly show that CaPif1 displays exclusively exonuclease activity and unambiguously establish the directionality of the exonuclease activity as the 3'-to-5' polarity. The enzyme appears to follow the two-metal-ion driven hydrolyzing activity exhibited by most of the nucleases, as shown by its dependence of magnesium and also by the identification of aspartic residues. Interestingly, an excellent correlation could be found between the presence of the conserved residues and the exonuclease activity when testing activities on Pif1 enzymes from eight fungal organisms. In contrast to others proteins endowed with the double helicase/exonuclease functionality, CaPif1 differs in the fact that the two activities are embedded in the same helicase domain and not located on separated domains. Our findings may suggest a biochemical basis for mechanistic studies of Pif1 family helicases.


Assuntos
Candida albicans/enzimologia , DNA Helicases/química , DNA Helicases/metabolismo , Exonucleases/metabolismo , Sequência de Aminoácidos , Candida albicans/química , Sequência Conservada , Exonucleases/química , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Hidrólise , Magnésio/metabolismo , Domínios Proteicos
5.
Nanomedicine ; 13(4): 1341-1351, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28115250

RESUMO

This project aimed to develop and characterize a new nanoadsorbent for hemoperfusion. Fe3O4 nanoparticles synthesized by a facile solvothermal method were coated with SiO2 and further modified by DMSA. TEM, XRD, FTIR, XPS and SEM were performed before and after lead adsorption to reveal the general performance and adsorption mechanism. Rabbit lead poisoning models were established to study the adsorption rate; then, a pig hemoperfusion experiment was used for further validation. In addition, coagulation, liver, kidney and heart function, blood lipids, electrolytes and the immune inflammatory system were studied before and after hemoperfusion. The results indicated that the materials had a high adsorption rate and chemisorbed lead mainly in the plasma. No obvious coagulation-fibrinolysis, organ toxicity, electrolyte disturbances, inflammatory reactions or immunosuppression was observed. The excellent blood compatibility and high biosafety of this material demonstrate its potential as a new type of hemoperfusion adsorbent.


Assuntos
Óxido Ferroso-Férrico/química , Hemoperfusão , Intoxicação por Chumbo/terapia , Chumbo/química , Nanopartículas/química , Adsorção , Animais , Teste de Materiais , Coelhos , Dióxido de Silício/química , Succímero/química , Suínos , Testes de Toxicidade
6.
Oncol Rep ; 35(5): 2599-605, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26934841

RESUMO

Cancer testis antigen sperm-associated antigen 9 (SPAG9) is highly expressed in many types of cancers. In the present study, to obtain a better understanding of the relevance of SPAG9 in cancer diagnosis and treatment, the expression of SPAG9 mRNA and protein in lung cancer specimens was evaluated by RT-PCR, western blotting and immunohistochemistry. ELISA was used to quantify the SPAG9 autoantibody in the peripheral blood of lung cancer patients. The results showed that the expression of SPAG9 mRNA and protein in the lung cancer tissues was significantly higher than that in the adjacent non-cancerous tissues (P<0.01). The level of the SPAG9 autoantibody in the serum of lung cancer patients was significantly higher than the level in the healthy controls (P<0.001), and the level of the SPAG9 autoantibody in the serum of untreated patients was significantly higher than that in treated patients (P=0.002). SPAG9 IgG antibody levels were significantly lower in treated adenocarcinoma and small cell lung cancer patients than these levels in the untreated patients (P=0.006, P=0.026, respectively), while no statistical difference was found between treated and untreated squamous cell carcinoma patients. Our results suggest that the SPAG9 antibody in serum is a promising marker for the diagnosis of lung cancer, and the level of the humoral immune response to this antigen appears to be related to the type of lung cancer.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/metabolismo , Neoplasias Pulmonares/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Adulto , Idoso , Autoanticorpos/sangue , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/mortalidade , Estudos de Casos e Controles , Feminino , Expressão Gênica , Humanos , Imunoglobulina G/sangue , Pulmão/metabolismo , Pulmão/patologia , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/mortalidade , Masculino , Pessoa de Meia-Idade
7.
Int J Clin Exp Med ; 8(8): 13928-36, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26550349

RESUMO

BACKGROUND/AIMS: To study the expression and clinical significance of serum soluble major histocompatibility complex class I-related chain A/B (sMICA/B), and its correlation with percentage of CD4(+), CD8(+), and NK cells, Liver fibrosis screening test, and liver enzymes in alcoholic liver disease (ALD). METHODS: Hainan Li ALD patients (n = 141) and healthy Li subjects (n = 100) were enrolled for the study. Liver enzymes were measured using automatic biochemical analyzer and Liver fibrosis screening test was used to study the correlation. In addition, sMICA/B expression in serum and percentage of CD4(+), CD8(+), and NK cells were determined using ELISA and flow cytometry respectively. RESULTS: Liver fibrosis screening test results and liver enzymes concentration were significantly higher (both P < 0.01), whereas the expression of sMICA and sMICB was significantly indifferent (P > 0.01) between ALD patients and healthy controls. However, percentage of CD4(+), CD8(+), and NK cells were statistically lower in ALD patients than in healthy controls. The Kendall's tau-b correlation coefficient for sMICA and sMICB/sMICA and LV was 0.561 and 0.120 respectively (P < 0.01). Pearson correlation coefficient of sMICA with the percentage of CD4(+), CD8(+)%, and NK cells was -0.587, -0.525, and -0.232 respectively, whereas the coefficient of sMICB was -0.590, -0.554, and -0.292 respectively (P < 0.01). CONCLUSION: 1. Liver fibrosis screening test is an excellent non-invasive approach for the diagnosis of hepatic fibrosis and shows significant correlation with liver enzymes. 2. sMICA and sMICB failed to assess the degree of hepatic fibrosis. 3. Decreased percentage of CD4(+), CD8(+), and NK cells were attributed as one of the risk factors for ALD.

8.
Int J Clin Exp Med ; 8(3): 4721-6, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26064412

RESUMO

This study's objectives are to assess the efficacy of detecting apoptotic caspase-3, -8, and -9 in human sperm and plasma using enzyme-linked immunosorbent assays (ELISA), and to compare these levels between fertile and infertile patient groups of Li nationality in China. This study offers a non-invasive, alternative strategy to analyzing sperm parameters in infertile males. Fifty-six infertile males were investigated; asthenospermia (n = 19), oligoasthenoteratozoospermia (n = 20), azoospermia (n = 17) compared with 20 healthy fertile controls. They were subjected to semen analysis by computer-assisted sperm assay (CASA). We found that caspase-3, -8, -9 existed in all specimens in both sperms and plasma. The level of caspase-3 and caspase-8 in plasma were both significantly higher than in sperm. Levels of caspase-8 and caspase-9 in sperm and plasma were significantly negatively correlated with sperm concentration, motility and A % (motility grade A). The level of caspase-8 in plasma was significantly negatively correlated with sperm concentration. However, only in healthy fertile controls sperm concentration was significantly negatively correlated with caspase-9 in sperm. Compared with the healthy fertile controls, only the OAT group exhibited significantly increased level of caspase-8 in sperm (P < 0.05). It is concluded that caspase-8 and caspase-9 in sperm and plasma are correlated with sperm motility, and can reflect the quality of sperm in vitro.

9.
Int J Clin Exp Med ; 8(2): 2760-5, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25932231

RESUMO

The present study was aimed to investigate the effects of the cytochrome P450 (CYP) 2D6*10 genetic polymorphism on postoperative patient-controlled morphine usage. A total of 114 patients were selected, and 102 patients completed the study. Polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) was used to determine the CYP2D6*10 genotype, and patients were categorized into three groups according to CYP2D6 genotype: heterozygous (m/w), wild-type homozygous (w/w), and mutant homozygous (m/m). Total morphine usage and visual analogue score (VAS) were determined 72 hours after the operation and compared across the three genotype groups. Statistical methods used to analyze results were the χ(2) test, analysis of variance, and multiple linear regression analysis; P<0.05 was considered to be statistically significant. The cumulative use of morphine in the m/w group was significantly higher than that in the m/m group between T0.5 and T4h (P<0.05). There were no significant differences in the loading dose of morphine or VAS among the different genotypes within 72 hours of operation. Patients carrying the CYP2D6*10 m/w genotype required higher doses of morphine at T0.5~T4h compared to the m/m group, and therefore received a higher cumulative dose of morphine post-operation. This phenomenon may be due to a decreased ability to synthesize endogenous opioid peptide.

10.
Int J Clin Exp Med ; 8(10): 19274-81, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26770564

RESUMO

OBJECTIVE: To investigate the presence of anti-sperm antibodies (AsAb) and the correlation between AsAb positivity and the expression of soluble major histocompatibility complex class I chain-related A and B (sMICA or sMICB) in the sera of infertile people of the Li nationality from Hainan, China. METHOD: A total of 136 people (68 couples) from five villages in the Wuzhishan region, Hainan province participated in this study. Among them, 31 couples were included in the fertile group and 37 couples in the infertile group. AsAb and sMICA/sMICB levels in serum were detected by ELISA. The median sMICA/sMICB levels between and among groups were compared by Mann-Whitney rank U testing and Kruskal-Wallis H testing, and the AsAb positivity rate was compared by Pearson Chi-Square testing. Correlation analysis was performed by calculating the Spearman's rho coefficient for nonparametric data. RESULTS: The serum levels for the fertile group (AsAb: 15.5 [4.0~127.0] U/ml, sMICA: 18.33 [13.30~52.40] pg/ml, sMICB: 27.72 [18.63~47.43] pg/ml) were not statistically different from those for the infertile group (AsAb: 18.0 [9.8~95.0] U/ml, sMICA: 20.95 [15.78~23.81] pg/ml, sMICB: 26.26 [18.06~61.38] pg/ml). However, grouping based on AsAb positivity revealed a statistically significant difference for the sMICA/sMICB levels (AsAb positive group: sMICA: 5.56 [4.30~17.23] pg/ml, sMICB: 16.13 [7.54~25.43] pg/ml; AsAb negative group: sMICA: 22.00 [18.05~66.13] pg/ml, sMICB: 36.51 [20.53~67.22] pg/ml; P < 0.01). These results suggest that AsAb is negatively associated with both sMICA (Spearman's coefficient, -0.475, P < 0.01) and sMICB (Spearman's coefficient, -0.381; P < 0.01). The analysis also shows that sMICA levels are positively associated with sMICB levels (Spearman's coefficient, 0.635; P < 0.01). CONCLUSION: AsAb can be detected in the serum of fertile and infertile Li people. However, there appears to be limited clinical value in the conventional detection of AsAb, sMICA and sMICB in serum for diagnosing infertility. People with positive AsAb expression have lower levels of sMICA/sMICB expression in serum, which may be one mechanism by which people produce AsAb.

11.
Cancer Epidemiol ; 36(6): 525-7, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22795327

RESUMO

BACKGROUND: Current evidence suggests that a majority of the inherited risks play a major role in glioma susceptibility, and glioma is due to the co-inheritance of multiple low-risk variants. These variants can be identified through association studies including such as genome-wide association studies (GWAS), which has led the glioma epidemiology researchers to focus on identifying potential disease-causing factors. METHODS: We evaluated and validated 10 tag single nucleotide polymorphisms (tSNPs) in seven genes associated with glioma susceptibility in a Han Chinese population, including 301 glioma cases and 302 controls, using a multiplexed single nucleotide polymorphism (SNP) MassEXTEND assay. We ascertained the genotypic frequencies for each tSNP in control subjects were within Hardy-Weinberg equilibrium (HWE) using an exact test, and then compared the genotype and allele frequencies of glioma patients and control subjects using the χ2 test. We then applied three genetic models (dominant, recessive, and additive) using PLINK software to assess the association of each tSNP with glioma risk. RESULTS: We identified two tSNPs to be associated with glioma susceptibility (rs1695, GSTP1, P = 0.019; rs2853676, TERT, P = 0.039), which we confirmed using dominant and additive model analyses. The genotype “GA” for rs1695 was recognized to be a protective genotype for glioma (OR, 0.67; 95% CI, 0.47-0.96; P = 0.027), while the genotype “AG” for rs2853676 was shown to be a risk genotype for glioma (OR, 1.50; 95% CI, 1.05-2.15; P = 0.025). CONCLUSION: Our results, and those from previous studies, suggest potential genetic contributes for GSTP1 and TERT in glioma development.


Assuntos
Neoplasias Encefálicas/genética , Glioma/genética , Glutationa S-Transferase pi/genética , Telomerase/genética , Adulto , Grupo com Ancestrais do Continente Asiático , Neoplasias Encefálicas/enzimologia , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Genótipo , Glioma/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Fatores de Risco
12.
Sheng Wu Gong Cheng Xue Bao ; 28(11): 1388-97, 2012 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-23457791

RESUMO

To establish a prokaryotic expression and purification protocol for nuclease P1 (NP1), we first obtained a synthetic NP1 by splicing 22 oligonucleotides with overlapping PCR. We constructed and transformed a secretory expression vector pMAL-p4X-NP1 into Escherichia coli host strains T7 Express and Origami B (DE3) separately. Then, the recombinant NP1 was purified by amylose affinity chromatography, and its activity, thermo-stability and metal-ion dependence were investigated systematically. The results indicated that the expressed fusion proteins MBP-NP1 (Maltose binding protein-NP1) existed mainly in soluble form both in host strains T7 Express and Origami B (DE3), but the specific activity of recombinant protein from Origami B(DE3) strain was higher than T7 Express strain (75.48 U/mg : 51.50 U/mg). When the MBP-tag was cleaved by protease Factor Xa, the specific activity both increased up to 258.1 U/mg and 139.2 U/mg. The thermal inactivation experiments demonstrated that the recombinant NP1 was quite stable, and it retained more than 90% of original activity after incubated for 30 min at 80 degrees C. Zn2+ (2.0 mmol/L) could increase enzyme activity (to 119.1%), on the contrary, the enzyme activity was reduced by 2.0 mmol/L Cu2+ (to 63.12%). This research realized the functional expression of NP1 in the prokaryotic system for the first time, and provided an alternative pathway for NP1 preparation.


Assuntos
Proteínas Fúngicas/biossíntese , Proteínas Fúngicas/metabolismo , Endonucleases Específicas para DNA e RNA de Cadeia Simples/biossíntese , Endonucleases Específicas para DNA e RNA de Cadeia Simples/metabolismo , Clonagem Molecular , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Fúngicas/genética , Genes Sintéticos , Vetores Genéticos/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Endonucleases Específicas para DNA e RNA de Cadeia Simples/genética
13.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue ; 15(3): 170-3, 2003 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-12831624

RESUMO

OBJECTIVE: To investigate the relationship of oxygen free radical (OFR) with per-oxidative injury of erythrocyte induced by intravenous procaine in vivo and the effect of methylene blue (MB) in removal of nitric oxide (NO) and peroxynitrite (ONOO(-)). METHODS: Forty patients undergoing elective surgery were divided randomly into intravenous procaine anesthesia (IPA) group and fentanyl group. Blood sample was taken before anesthesia (T0), 120 minutes (T1) and 180 minutes (T2) after IPA and 30 minutes after treatment with MB (1-2 mg/kg, T3) to determine the changes in the levels of NO, OFR, lipid peroxide (LPO), superoxide dismutase (SOD), catalase (CAT), NADH-Cyt b5-reductase (Cyt b5-R) and methemoglobin (MHb). RESULTS: Compared with T0, the levels of NO, OFR, LPO, MHb in IPA group were significantly increased at T1,T2. At same time SOD, CAT and Cyt b5-R were significantly decreased. NO, OFR, MHb, SOD, CAT and Cyt b5-R were all reduced to the normal levels at T3. No changes in any determined parameters in fentanyl group during anesthesia. CONCLUSION: It is indicated that the metabolites of procaine consist of a large quantity of NO:ONOO(-), producing per-oxidative injury to erythrocyte. MB is effective in eliminating OFR in vivo, protecting tissue cells. It may act as an antioxidant drug in the treatment of critical illness.


Assuntos
Antioxidantes/farmacologia , Azul de Metileno/farmacologia , Adolescente , Adulto , Idoso , Catalase/sangue , Catalase/efeitos dos fármacos , Feminino , Humanos , Peróxidos Lipídicos/sangue , Masculino , Metemoglobina/efeitos dos fármacos , Metemoglobina/metabolismo , Pessoa de Meia-Idade , NADPH-Ferri-Hemoproteína Redutase/sangue , NADPH-Ferri-Hemoproteína Redutase/efeitos dos fármacos , Óxido Nítrico/sangue , Ácido Peroxinitroso/sangue , Espécies Reativas de Oxigênio/sangue , Superóxido Dismutase/sangue , Superóxido Dismutase/efeitos dos fármacos
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