ABSTRACT
SO2, a gas signaling molecule, can be produced endogenously in mitochondria. Its hydrolysate, HSO3-, plays a key role in food preservation, cardiovascular relaxation, and other fields, suggesting that it is important to achieve its detection. Here, based on the Michael addition mechanism, four hemicyanine dye fluorescent probes (ETN, ETB, STB, and EIB) were designed and synthesized for responding to HSO3-. We evaluated the reaction ability of different probes with HSO3- and tried to explain the reasons for the significantly different response effects between probes and HSO3- according to the structure-activity relationship. The influence of different substituents of probes on the properties of mitochondria-targeting was also discussed. Finally, we screened out ETN as the optimal HSO3- probe due to its high sensitivity, rapid reactivity, and good mitochondria-targeting, and it could sensitively respond to HSO3- in living cells. The LODs of ETN for HSO3- were calculated by both absorption and fluorescence methods, respectively, which were 2.727 and 0.823 µM. Our work provided valuable references for designing strategies and potential tools for response to SO2 derivatives in biosystems.
Subject(s)
Fluorescent Dyes , Mitochondria , Humans , Carbocyanines , Limit of Detection , Sulfites , HeLa CellsABSTRACT
The G-quadruplex ligands database (G4LDB, http://www.g4ldb.org) provides a unique collection of reported G-quadruplex ligands to streamline ligand/drug discovery targeting G-quadruplexes. G-quadruplexes are guanine-rich nucleic acid sequences in human telomeres and gene promoter regions. There is a growing recognition for their profound roles in a wide spectrum of diseases, such as cancer, diabetes and cardiovascular disease. Ligands that affect the structure and activity of G-quadruplexes can shed light on the search for G-quadruplex-targeting drugs. Therefore, we built the G4LDB to (i) compile a data set covering various physical properties and 3D structure of G-quadruplex ligands; (ii) provide Web-based tools for G-quadruplex ligand design; and (iii) to facilitate the discovery of novel therapeutic and diagnostic agents targeting G-quadruplexes. G4LDB currently contains >800 G-quadruplex ligands with â¼4000 activity records, which, to our knowledge, is the most extensive collection of its kind. It offers a user friendly interface that can meet a variety of data inquiries from researchers. For example, ligands can be searched for by name, molecular properties, structures, ligand activities and so on. Building on the reported data, the database also provides an online ligand design module that can predict ligand binding affinity in real time.
Subject(s)
Databases, Chemical , Drug Design , G-Quadruplexes/drug effects , Internet , Ligands , Molecular Docking SimulationABSTRACT
Oxygen is one of the determinants of root microbiome formation. However, whether plants regulate rhizosphere oxygen levels to affect microbiota composition and the underlying molecular mechanisms remain elusive. The receptor-like kinase (RLK) family member FERONIA modulates the growth-defense tradeoff in Arabidopsis. Here, we established that rice FERONIA-like RLK 7 (FLR7) controls rhizosphere oxygen levels by methylene blue staining, oxygen flux, and potential measurements. The formation of oxygen-transporting aerenchyma in roots is negatively regulated by FLR7. We further characterized the root microbiota of 11 FLR mutants including flr7 and wild-type Nipponbare (Nip) grown in the field by 16S ribosomal RNA gene profiling and demonstrated that the 11 FLRs are involved in regulating rice root microbiome formation. The most abundant anaerobic-dependent genus Anaeromyxobacter in the Nip root microbiota was less abundant in the root microbiota of all these mutants, and this contributed the most to the community differences between most mutants and Nip. Metagenomic sequencing revealed that flr7 increases aerobic respiration and decreases anaerobic respiration in the root microbiome. Finally, we showed that a representative Anaeromyxobacter strain improved submergence tolerance in rice via FLR7. Collectively, our findings indicate that FLR7 mediates changes in rhizosphere oxygen levels and enriches the beneficial dominant genus Anaeromyxobacter and may provide insights for developing plant flood prevention strategies via the use of environment-specific functional soil microorganisms.
Subject(s)
Bacteria , Oryza , Bacteria/genetics , Rhizosphere , Plant Roots/genetics , Soil Microbiology , SoilABSTRACT
Enalaprilat (Ena.), an angiotensin II (Ang II) converting enzyme inhibitor (ACEI), can produce some therapeutic effects on hypertension, ventricular hypertrophy and myocardial remodeling in clinic, but its precise mechanism, especially its signaling pathways remain elusive. In this study, cardiac fibroblasts (CFb) was isolated by the trypsin digestion method; a BrdU proliferation assay was adopted to determine cell proliferation; an immunofluorescence assay was used to measure intracellular reactive oxygen species (ROS); immunocytochemistry staining and Western blotting assay were used to detect phosphorylated p38 mitogen activated protein kinase (p-p38MAPK) and transforming growth factor-ß(1) (TGF-ß(1)) protein expression, respectively. The results showed that Ang II (10(-7) M) stimulated the cardiac fibroblast proliferation which was inhibited by NAC (an antioxidant), SB203580 (a p38MAPK inhibitor) or enalaprilat; Ang II caused an burst of intracellular ROS level within thirty minutes, an increase in p-p38MAPK (3.6-fold of that in the control group), as well as an elevation of TGF-ß(1) meantime; NAC, an antioxidant, and enalaprilat treatment attenuated cardiac fibroblast proliferation induced by Ang II and decreased ROS and p-p38MAPK protein levels in rat cardiac fibroblast; SB203580 lowered TGF-ß(1) protein expression in rats' CFb in a dose-dependent manner. It could be concluded that enalaprilat can inhibit the cardiac fibroblast proliferation induced by Ang II via blocking ROS/P38MAPK/TGF-ß(1) signaling pathways and the study provides a theoretical proof for the application of ACEIs in treating myocardial fibrosis and discovering the primary mechanism through which ACEIs inhibit CFb proliferation.
Subject(s)
Angiotensin-Converting Enzyme Inhibitors/pharmacology , Cell Proliferation/drug effects , Enalaprilat/pharmacology , Fibroblasts/physiology , MAP Kinase Signaling System , Myocardium/cytology , Acetylcysteine/pharmacology , Active Transport, Cell Nucleus/drug effects , Angiotensin II/physiology , Animals , Cell Survival/drug effects , Cells, Cultured , Fibroblasts/drug effects , Gene Expression/drug effects , Imidazoles/pharmacology , Phenols/pharmacology , Phosphorylation/drug effects , Plant Extracts/pharmacology , Pyridines/pharmacology , Rats , Rats, Inbred WKY , Reactive Oxygen Species/metabolism , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
OBJECTIVE: To observe the predictability of corneal flap creation with the FEMTO LDV femtosecond laser and analyze preliminarily the factors correlating to the thickness and diameter of the flap . METHODS: It was a study of serial cases. 260 eyes of 130 consecutive patients were treated with the FEMTO LDV. The eyes were assigned to two groups according to intended flap thickness, 110 µm (208 eyes) and 90 µm (52 eyes). Intended flap diameter varied from 8.5 to 9.5 mm. Difference analysis of flap diameter and intended diameter as well as flap thickness and intended thickness were made. The data was analyzed with SPSS to sum up a multiple stepwise regression formula that could express their quantitative relationship. RESULTS: The 90 µm flap group had a average flap thickness of (95.12 ± 7.65) µm, while for the 110 µm group the average flap thickness was (104.81 ± 3.09) µm. The difference between right and left eyes was not statistically significant (t(110 µm) = -1.223, t(90 µm) = -1.343, P > 0.05). Corneal flap thickness was inversely correlated with flap diameter (r(110 µm) = -0.143, r(90 µm) = -0.315, P < 0.05), but was not related to preoperative patient age, corneal thickness, keratometric value K or intraocular pressure (r(110 µm) = -0.160, 0.054, -0.011, -0.363; r(90 µm) = 0.024, 0.074, -0.212, -0.434, all P > 0.05). Corneal flap diameter was positively correlated with preoperative corneal keratometric value K and thickness (P < 0.001, P < 0.05). Multiple stepwise regression analysis showed flap diameter was an influencing factor for flap thickness. Preoperative corneal keratometric value K and thickness were influencing factors for flap diameter. CONCLUSIONS: The LASIK flap creation with the FEMTO LDV laser has relatively good predictability. Flap diameter is an influencing factor for flap thickness.
Subject(s)
Cornea/surgery , Keratomileusis, Laser In Situ/methods , Myopia/surgery , Surgical Flaps , Adolescent , Adult , Cornea/anatomy & histology , Corneal Transplantation , Female , Humans , Male , Young AdultABSTRACT
BACKGROUND: In mirror-image dextrocardia, the anterior-posterior position of the cardiac chambers and great vessels is maintained, but the left-right orientation of the abdominal organs is reversed. The abnormal anatomy of the heart poses surgical challenges and problems in dealing with surgical risk and monitoring complications. There are few reports on closure of the left atrial appendage (LAA) in dextrocardia and no reports on the application of enhanced recovery after surgery (ERAS) following LAA occlusion (LAAO) procedures. CASE SUMMARY: The objective for this case was to ensure perioperative safety and accelerate postoperative recovery from LAAO in a patient with mirror-image dextrocardia. ERAS was guided by the theory and practice of nursing care. Atrial fibrillation was diagnosed in a 77-year-old male patient, in whom LAAO was performed. The 2019 guidelines for perioperative care after cardiac surgery recommend that the clinical nursing procedures for patients with LAAO should be optimized to reduce the incidence of perioperative complications and ensure patient safety. Music therapy can be used throughout perioperative treatment and nursing to improve the anxiety symptoms of patients. CONCLUSION: The procedure was uneventful and proceeded without complications. Anxiety symptoms were improved.
ABSTRACT
OBJECTIVE: Gene polymorphism is closely related to tumor development, therapeutic response and prognosis. The relationship between regenerating gene 1A (Reg1A) polymorphism and nasopharyngeal carcinoma (NPC) is unclear. This retrospective study aimed to analyze the association between Reg1a polymorphisms and metastasis, radiation sensitivity and survivals in patients with NPC. METHODS: A total of 308 patients who had received radiotherapy at the Affiliated Xinhua Hospital, Hainan Medical College, between January 2010 and December 2018 with NPC, were enrolled for assessment of Reg1a polymorphisms through direct DNA sequencing. RESULTS: In the polymorphism of gene REG1A, patients with rs10165462 20CC genotype had later T stages (OR = 4.051, 95% CI: 1.775-9.244, P = 0.001), whereas carriers with rs12072 2922CC genotype had earlier T stages (OR = 1.891, 95% CI: 1.018-3.514, P = 0.044) after adjustments for age and gender, respectively. Among rs10165462 20 C/T polymorphism, 20TT wild-type was associated with better radiation response (P = 0.0019), and multivariate analysis showed that it was the only genotype of polymorphism that was significantly associated with better radiation response (OR = 0.265, 95% CI: 0.096-0.727, P = 0.01). Patients with the 20TT wild-type had a better five-year overall survival (60.9%) rate and five-year progression-free survival (60.8%) than those with the 20CC genotype (41.8% and 39.4%, P = 0.01 and P = 0.004, respectively). Patients with variant alleles (CC + CT) had significantly poorer OS (45.2%) and PFS (41.8%) compared with wild-type (TT) carriers (60.9% and 60.8%; P = 0.037 and P = 0.015, respectively). As for rs12072, patients with variant alleles (TT + TC) had significantly adverse OS and PFS compared with wild-type (CC) carriers (62.5% vs 44.8% and 62.5% vs 42.9%; P = 0.024 and P = 0.027, respectively). Cox regression showed that rs10165462 20CT was the only prognostic factor for OS (HR = 1.642, 95% CI 1.038-2.598, P = 0.034) and PFS (HR = 1.705, 95% CI 1.080-2.692, P = 0.022). CONCLUSION: Reg1a polymorphisms may be a predictor of radiation response, local invasion, OS and PFS in patients with NPC who undergo radiotherapy treatment.
ABSTRACT
A single-stranded G-tract human telomere DNA sequence is able to fold into intramolecular G-quadruplex structures which may be important for a number of biological processes and disease-related mechanisms. Poly(L-lysine) (PLL) polymer is linear polypeptides with lysine as the repeat unit and has been employed as a gene carrier in achieving targeted delivery of DNA to cancer cells. To explore the influence of PLL on the conformation of Hum24 DNA, we have investigated the interaction of PLL with Hum24 by biophysical methods, mainly CD, ESI-MS, and polyacrylamide gel electrophoresis for the first time. The CD data have shown that PLL can induce single-stranded Hum24 to form an intramolecular parallel G-quadruplex structure, further confirmed by ESI-MS analysis and gel electrophoresis results. The formation of an intramolecular G-quadruplex is strongly dependent on the Hum24/PLL molar ratios and the length of both the polypeptides and oligonucleotide. Such phenomena may be interpreted by electrostatically attracting negative-charged Hum24 by positive-charged PLL which facilitates the close contact between the guanines and formation of hydrogen bonding, thus leading the final shape of a G-quadruplex structure.
Subject(s)
DNA, Single-Stranded/chemistry , G-Quadruplexes , Polylysine/chemistry , Potassium/chemistry , Sodium/chemistry , Telomere/chemistry , Circular Dichroism , Electrophoresis, Polyacrylamide Gel , Humans , Nucleic Acid Conformation , Oligodeoxyribonucleotides/chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
AIM: To investigate the anti-inflammatory mechanism of the polysaccharides of Ginkgo biloba leaves (PGBL) by inhibiting leucocyte adhesion. METHODS: The rough PGBL were isolated and purified. The anti-inflammatory effects of purified PGBL (p-PGBL) were assayed by ear edema induced by xylol and the acute peritonitis model in mice. The effect of p-PGBL on inhibiting the interaction between P-selectin and its ligands was investigated by flow cytometry and flow chamber. RESULTS: p-PGBL could effectively inhibit the acute inflammation in mice and interfere with the adhesion of HL-60 cells, a human leukaemia cell line, or neutrophils to P-selectin in static conditions, as well as the adhesion of neutrophils to Chinese hamster ovary cells expressing human P-selectin and human umbilical vein endothelial cells in flow conditions in a dose-dependant manner. CONCLUSIONS: p-PGBL can inhibit the inflammatory process through interfering with the interaction between P-selectin and its ligands.
Subject(s)
Ginkgo biloba/immunology , Inflammation/immunology , Neutrophils/drug effects , P-Selectin/immunology , Plant Leaves/immunology , Polysaccharides/pharmacology , Animals , CHO Cells , Cell Adhesion/drug effects , Cricetinae , Cricetulus , Dose-Response Relationship, Drug , Endothelium, Vascular/drug effects , Ginkgo biloba/metabolism , HL-60 Cells , Humans , Inflammation/metabolism , Mice , Mice, Inbred BALB C , P-Selectin/metabolism , Plant Leaves/metabolism , Polysaccharides/isolation & purification , Umbilical Veins/cytologyABSTRACT
The SNPs in partial coding sequence of MC3R and MC4R genes were identified by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) analysis and DNA sequencing in shack-Kee and Columba domestica from Harbin area. Correlation analysis between MC3R and MC4R polymorphism and growth and body composition traits was carried by the least square analysis. The genotypes of T91G mutation in MC3R gene and A903G mutation in MC4R gene proved to have significant association with body weight, carcass weight, and holo-carcass weight in shack-Kee (P<0.05). The interaction of MC3R-T91G and MC4R-A903G was discussed through combination genotype analysis. The least square analysis showed that the combined genotype had significant association with holo-carcass weight (P<0.05). Multiple comparisons revealed that BBAA genotype birds had a higher holo-carcass weight than AABB genotype birds and BBAA genotype was the beneficial genotype for the growth of body weight.
Subject(s)
Columbidae , Polymorphism, Single-Stranded Conformational , Animals , Body Weight/genetics , Genotype , Polymerase Chain Reaction , Polymorphism, Single NucleotideABSTRACT
Nucleic acid based molecular device is a developing research field which attracts great interests in material for building machinelike nanodevices. G-quadruplex, as a new type of DNA secondary structures, can be harnessed to construct molecular device owing to its rich structural polymorphism. Herein, we developed a switching system based on G-quadruplexes and methylazacalix[6]pyridine (MACP6). The induced circular dichroism (CD) signal of MACP6 was used to monitor the switch controlled by temperature or pH value. Furthermore, the CD titration, Job-plot, variable temperature CD and (1)H-NMR experiments not only confirmed the binding mode between MACP6 and G-quadruplex, but also explained the difference switching effect of MACP6 and various G-quadruplexes. The established strategy has the potential to be used as the chiral probe for specific G-quadruplex recognition.
Subject(s)
Calixarenes/chemistry , Computers, Molecular , DNA/chemistry , G-Quadruplexes , Oligonucleotides/chemistry , Circular Dichroism , DNA-Binding Proteins/chemistry , Hydrogen-Ion Concentration , Models, Molecular , Molecular Docking Simulation , Nuclear Magnetic Resonance, Biomolecular , TemperatureABSTRACT
Increasing evidence places Schisandrin B (Sch B) at an important position in nerve protection, indicating that Sch B might play a positive role in the therapy of neurodegenerative diseases. However, there is little information on it. Our studies showed that pretreatment with Sch B could reduce lactate dehydrogenase, malondialdehyde, and reactive oxygen species release and significantly increase the cell viability and the superoxide dismutase level. Sch B (10 µM) markedly inhibited cell apoptosis, whereas LY294002 (20 µM), a phosphatidylinositol-3 kinase inhibitor, blocked the antiapoptotic effect. More importantly, Sch B (10 µM) increased the phosphoprotein kinase B/protein kinase B (Akt) and B-cell lymphoma-2/Bcl-2 associated X protein ratios on preincubation with cells for 2 h, which was then inhibited by LY294002 (20 µM). Results indicate that Sch B can protect PC12 cells from apoptosis by activating the phosphatidylinositol-3 kinase/Akt signaling pathway and may emerge as a potential drug for neurodegenerative diseases.
Subject(s)
Apoptosis/drug effects , Lignans/pharmacology , Neurodegenerative Diseases/metabolism , Oxidative Stress/drug effects , Polycyclic Compounds/pharmacology , Animals , Cell Survival/drug effects , Cyclooctanes/pharmacology , Cyclooctanes/therapeutic use , Lignans/therapeutic use , Neurodegenerative Diseases/drug therapy , PC12 Cells , Polycyclic Compounds/therapeutic use , Rats , Reactive Oxygen Species/metabolism , Signal Transduction/drug effectsABSTRACT
AIM: To investigated the effects of the SNPs (rs411021, rs436045, rs427811, rs385039 and rs809912) on gene expression and further identify the susceptibility genes of type 2 diabetes. METHODS: Ten allele fragments (49 bp each) were synthesized according to the 5 SNPs mentioned above. These fragments were cloned into luciferase reporter gene vector and then transfected into HepG2 cells. The activity of the luciferase was assayed. Effects of the SNPs on RNA splicing were analyzed by bioinformatics. RESULTS: rs427811T allele and rs809912G allele enhanced the activity of the reporter gene expression. None of the 5 SNPs affected RNA splicing. CONCLUSION: SNPs in protein kinase Cz (PKCZ) gene probably play a role in the susceptibility to type 2 diabetes by affecting the expression level of the relevant genes.
Subject(s)
Gene Expression Regulation, Enzymologic/genetics , Luciferases/genetics , Polymorphism, Single Nucleotide/genetics , Protein Kinase C/genetics , Base Sequence , DNA Primers , Diabetes Mellitus, Type 2/enzymology , Diabetes Mellitus, Type 2/genetics , Exons/genetics , Genes, Reporter , Genetic Predisposition to Disease/genetics , Humans , Introns/genetics , Transcription, Genetic , TransfectionABSTRACT
AIM: To identify the susceptible gene (s) for type 2 diabetes in the previously mapped region, 1p36.33-p36.23, in Han population of North China using single nucleotide polymorphisms (SNPs) and to analyze the haplotypes of the gene (s) related to type 2 diabetes. METHODS: Twenty three SNPs located in 10 candidate genes in the mapped region were chosen from public SNP domains with bioinformatic methods, and the single base extension (SBE) method was used to genotype the loci for 192 sporadic type 2 diabetes patients and 172 normal individuals, all with Han ethical origin, to perform this case-control study. The haplotypes with significant difference in the gene (s) were further analyzed. RESULTS: Among the 23 SNPs, 8 were found to be common in Chinese Han population. Allele frequency of one SNP, rs436045 in the protein kinase C/zetagene (PRKCZ) was statistically different between the case and control groups(P<0.05). Furthermore, haplotypes at five SNP sites of PRKCZ gene were identified. CONCLUSION: PRKCZ gene may be associated with type 2 diabetes in Han population in North China. The haplotypes at five SNP sites in this gene may be responsible for this association.
Subject(s)
Asian People/genetics , Diabetes Mellitus, Type 2/genetics , Haplotypes , Protein Kinase C/genetics , Case-Control Studies , China/ethnology , Gene Frequency , Humans , Polymorphism, Single NucleotideABSTRACT
OBJECTIVE: To investigate the distribution of single nucleotide polymorphisms (SNPs) in CAPN10 gene in Chinese population and their relation with type 2 diabetes mellitus in Han people of Northern China. METHODS: CAPN10 gene was sequenced to detect SNPs in different nationalities of China. Five SNPs were chosen to perform case-control study and haplotype analysis in 156 normal Han people of Northern China and 173 type 2 diabetes. One SNP was also analyzed with transmission-disequilibrium test (TDT) and sib transmission-disequilibrium test (STDT) in 68 type 2 diabetes pedigrees (377 people). RESULTS: A total of 40 SNPs were identified in length of 8,936 bp, with an average of 1 in every 223 bp. The SNPs in CAPN10 gene did not distribute evenly and the SNPs in Chinese were different from those reported in Mexican American. There was no significantly statistical difference in the allele frequency of the 5 SNPs between case and control, and the haplotype frequencies in the two groups were not significantly different. No positive results was found in TDT and STDT analysis. CONCLUSIONS: The SNP distribution of CAPN10 gene differs in different nationalities. The studied SNPs in CAPN10 gene may not be the major susceptibility ones of type 2 diabetes mellitus in Han people of Northern China.
Subject(s)
Calpain/genetics , Diabetes Mellitus, Type 2/ethnology , Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Case-Control Studies , China , Ethnicity , Humans , Polymerase Chain ReactionABSTRACT
OBJECTIVE: To observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues. METHODS: Using serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI. RESULTS: A total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively. CONCLUSIONS: Genes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes.
Subject(s)
Adenoma/genetics , Gene Expression Profiling/methods , Pituitary Gland/metabolism , Pituitary Neoplasms/genetics , Adenoma/metabolism , Base Sequence , Cloning, Molecular , Expressed Sequence Tags , Gene Expression , Gene Expression Regulation, Neoplastic , Gene Library , Humans , Molecular Sequence Data , Oligonucleotide Array Sequence Analysis , Pituitary Neoplasms/metabolismABSTRACT
OBJECTIVE: To probe the candidate susceptibility gene (s) of type 2 diabetes in the formal mapping region, 1p36.33-p36.23, in Han people of Northern China using single nucleotide polymorphisms (SNPs). METHODS: 23 SNPs located in 10 candidate genes in the mapping region were chosen from public SNP domain by bioinformatic methods and single base extension (SBE) method were used to genotype the loci in 192 sporadic type 2 diabetes patients and 172 normal individuals to perform case-control study. RESULTS: Among the 23 SNPs, 8 were found to be common in Chinese population. There were statistically different in the allele frequency of 2 SNP, rs436045 in the protein kinase C/xi gene and rs228648 in Urotensin II gene between case and control groups. CONCLUSIONS: The two SNP may be associated with type 2 diabetes in Han people of China, which makes base for further study of the relation between the genes they located with type 2 diabetes.
Subject(s)
Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Protein Kinase C/genetics , Urotensins/genetics , Alleles , Case-Control Studies , Ethnicity , Genetic Testing , Genotype , HumansABSTRACT
OBJECTIVE: To investigate the distribution of the single nucleotide polymorphisms (SNPs) in CAPN10 gene in Chinese population and their relation with type 2 diabetes mellitus in Han people of Northern China. METHODS: CAPN10 gene was sequenced to detect SNPs in 27 samples of different nationalities in China. 5 SNPs were genotyped with single-base extension (SBE) method to perform case-control study in 156 normal Han people of Northern China and 173 type 2 diabetes and the 3 positive loci reported in the article were performed haplotype analysis. One positive locus was also analyzed with transmission-disequilibrium test (TDT) and sib transmission-disequilibrium test (STDT) in 68 type 2 diabetes pedigrees (377 cases). RESULTS: A total of 40 SNPs were identified in length of 8,936 bp, with an average of 1 in every 223 bp; The SNPs in CAPN10 gene did not distribute evenly and the SNPs in Chinese was different from that reported in American Mexicans. There was no significantly statistical difference in the allele frequency of the 5 SNPs between case and control (P > 0.05), and the haplotype frequencies in the two groups were not much different (P > 0.05). There was no positive results in TDT and STDT analysis (P > 0.05). CONCLUSIONS: The SNP distribution of CAPN10 gene varies with different nationalities. The studied SNPs in CAPN10 gene may not be the major susceptibility ones of type 2 diabetes mellitus in Han people of Northern China.
Subject(s)
Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Alleles , Asian People , Case-Control Studies , Ethnicity , Genetic Testing , Genotype , HumansABSTRACT
OBJECTIVES: To confirm previous whole-genome scan results of mapping type 2 diabetes susceptibility genes in chromosome 1 in Northern Chinese Han population by conducting a new genome scan with both an enlarged number of type 2 diabetes families and a new set of microsatellite markers. METHODS: A genome scan method was applied. After multiplexed PCR, electrophoreses, genescan and genotyping analysis, size informations for all loci were obtained, and a further study was done using both parametric and non-parametric linkage analysis to calculate the P-values and Z-values of these loci. RESULTS: A total of 34 microsatellite markers distributed within 5 regions along chromosome 1 were surveyed, and 12,000 genotypes were screened. Evidence of linkage with diabetes was identified for 8 of the 34 loci (all the P-values of the 8 loci distributed in 3 regions were lower than 0.05, and the highest Z-value was 2.17). Interestingly, all the 5 markers at the P terminal 1p36.3-1p36.23 region, spanning a long range of 16.9 cM, suggested to be linked with the disease. The results of the other two regions were not consistent with the previous ones. CONCLUSIONS: The study results have confirmed those gained in the previous genome-wide scan. The fact that all 5 loci at the P terminal region displayed linkage with diabetes suggests that more than 1 susceptibility gene may reside in this region.
Subject(s)
Chromosomes, Human, Pair 1 , Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Microsatellite Repeats/genetics , Asian People , Chromosome Mapping , Ethnicity , Genetic Linkage , Genetic Testing , Genotype , HumansABSTRACT
OBJECTIVE: To study the function of 5 single nucleotide polymorphisms (SNPs) of the PRKCZ gene, a susceptibility gene for type 2 diabetes in Han population of North China, in the pathogenesis of the disease. METHODS: Bioinformatic methods and reporter gene activity determination were used to analyze the function of the 5 SNPs. RESULTS: The reporter gene activities of different alleles of 2 SNPs, rs427811 and rs809912, were obviously different, which implies that these 2 SNPs might be susceptibility loci of the disease. CONCLUSION: The PRKCZ gene is further confirmed to be a susceptibility gene for type 2 diabetes in Han population of North China. Two SNPs in the gene play a role in the pathogenesis of the disease by affecting the expression level of PRKCZ gene.