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The micronucleomics test can comprehensively display a variety of harmful endpoints, such as DNA damage and repair, chromosome breakage or loss and cell growth inhibition, with fast, simple and economical feature. Micronucleomics is not only widely used in the comprehensive assessment of the types and modes of genetic action of exogenous chemicals (such as drugs, food additives, cosmetics, environmental pollutants, etc.), but also plays an important role in the screening and risk assessment of cancer population at high risk. However, the traditional micronucleomics image counting method has the characteristics of time-consuming, low accuracy, and high cost, which cannot meet the current analysis requirements of large-scale, multi-index, rapidity, high precision and visualization. In recent years, with the rapid development of the era of precision medicine based on big data, visualized analysis of new micronucleomics based on machine learning and detection strategies based on deep learning have shown a good application prospect. This review, based on the application value of micronucleomics, systematically compares the traditional and new artificial intelligence counting of micronucleus images, and discusses the future direction of micronucleus image detection.
Subject(s)
Humans , Artificial Intelligence , Big Data , Machine Learning , Precision MedicineABSTRACT
Objective: To compare the predictive value of HAS-BLED, HEMORR2HAGES, ATRIA and ORBIT scores on the bleeding risk in nonvalvular atrial fibrillation (NVAF) patients treated with dabigatran. Methods: Data of 942 NVAF patients participating a non-interventional prospective study of anticoagulant therapy with dabigatran, which was conducted in 12 centers from February 2015 to December 2017 in China, were analyzed. Complete HAS-BLED HEMORR2HAGES, ATRIA and ORBIT bleeding risk scores data and follow-up data were available in the enrolled patients. The endpoint of the study was bleeding events occurred during a 6 months follow-up. Cox proportional hazards models were constructed to analyze the associations between HAS-BLED, HEMORR2HAGES, ATRIA and ORBIT scores and risk of bleeding, and the area under the curve (AUC) of receiver operating characteristics curves (ROC) of each score was used to set the predictive value for bleeding risk. Results: Among the 942 patients, the mean age was (65.3±11.2) years old, 542 (57.5%) were males. A total of 93 (9.9%) bleeding events occurred during follow up, 89 (9.4%) events were minor bleeding, and 4 (0.4%) events were major bleeding. Patients with a high-risk HAS-BLED score had a 1.87-fold increased risk of bleeding compared with low-risk patients (HR = 2.87, 95% CI:1.26-6.51, P = 0.012). There was no statistically significant difference between low-medium-high-risk grading in other scoring systems and bleeding risk (all P>0.05). The AUC (95%CI) of HAS-BLED, HEMORR2HAGES, ATRIA and ORBIT bleeding risk scores were 0.558 (0.525-0.590), 0.520 (0.487-0.553), 0.513(0.480-0.545), 0.523(0.490-0.555), respectively. The AUC of all bleeding score systems were of ≤ 0.700. Conclusion: Among the NVAF patients taking dabigatran in China, the HAS-BLED bleeding risk score is superior to other 3 bleeding risk score on predicting the bleeding risk in these patients, but its predictive value is still relatively low.
Subject(s)
Aged , Humans , Male , Middle Aged , Anticoagulants , Atrial Fibrillation , China , Dabigatran , Prospective Studies , Risk Assessment , Risk Factors , StrokeABSTRACT
BACKGROUND@#The association between peripheral leukocyte count and bleeding events in nonvalvular atrial fibrillation (NVAF) patients treated with dabigatran remains unclear. This study aimed to explore the association between leukocyte count and bleeding events after excluding other confounders in NVAF patients taking dabigatran.@*METHODS@#A total of 851 NVAF patients treated with dabigatran (110 mg bid) were recruited from 12 centers in China from February 2015 to December 2017. Follow-up was completed by May 2018. The exposure and outcome variables were leukocyte count measured at baseline and the number of bleeding events within the subsequent 6 months. Multivariate Cox proportional hazards models were constructed to analyze independent associations, and a Cox proportional hazards regression with cubic spline functions and smooth curve fitting (penalized spline method) was used to address nonlinearity between leukocyte count and bleeding. The inflection point was calculated using a recursive algorithm, and then a two-piecewise Cox proportional hazards model for both sides of the inflection point was constructed.@*RESULTS@#During 6-month follow-up, 87 participants occurred bleeding events. For every 1 × 10/L increase in leukocyte count, the risk of bleeding increased by 11% (hazard ratio [HR]: 1.11, 95% confidence interval [CI]: 0.99-1.25). The smooth curve showed nonlinear relationship between leukocyte count and bleeding events. The inflection point of the leukocyte count was 6.75 × 10/L. For leukocyte counts < 6.75 × 10/L, the HR (95% CI) was 0.88 (0.69-1.13), and for leukocyte counts ≥ 6.75 × 10/L, the HR (95% CI) was 1.28 (1.09-1.51).@*CONCLUSION@#This study found a J-shaped association between baseline leukocyte count and risk of bleeding in NVAF patients treated with dabigatran.@*CLINICAL TRIAL REGISTRATION@#NCT02414035, https://clinicaltrials.gov.
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Background@#The association between peripheral leukocyte count and bleeding events in nonvalvular atrial fibrillation (NVAF) patients treated with dabigatran remains unclear. This study aimed to explore the association between leukocyte count and bleeding events after excluding other confounders in NVAF patients taking dabigatran.@*Methods@#A total of 851 NVAF patients treated with dabigatran (110 mg bid) were recruited from 12 centers in China from February 2015 to December 2017. Follow-up was completed by May 2018. The exposure and outcome variables were leukocyte count measured at baseline and the number of bleeding events within the subsequent 6 months. Multivariate Cox proportional hazards models were constructed to analyze independent associations, and a Cox proportional hazards regression with cubic spline functions and smooth curve fitting (penalized spline method) was used to address nonlinearity between leukocyte count and bleeding. The inflection point was calculated using a recursive algorithm, and then a two-piecewise Cox proportional hazards model for both sides of the inflection point was constructed.@*Results@#During 6-month follow-up, 87 participants occurred bleeding events. For every 1 × 109/L increase in leukocyte count, the risk of bleeding increased by 11% (hazard ratio [HR]: 1.11, 95% confidence interval [CI]: 0.99–1.25). The smooth curve showed nonlinear relationship between leukocyte count and bleeding events. The inflection point of the leukocyte count was 6.75 × 109/L. For leukocyte counts < 6.75 × 109/L, the HR (95% CI) was 0.88 (0.69–1.13), and for leukocyte counts ≥ 6.75 × 109/L, the HR (95% CI) was 1.28 (1.09–1.51).@*Conclusion@#This study found a J-shaped association between baseline leukocyte count and risk of bleeding in NVAF patients treated with dabigatran.@*Clinical trial registration@#NCT02414035, https://clinicaltrials.gov.
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·Long non-coding RNAs (lncRNAs) refer to a kind of non-coding RNA which is longer than 200 nucleotides, with the characteristic of its numerous, diversity of types and modes of action. The biological functions of lncRNA involved in genomic imprinting, chromatin remodeling, translational control of mRNA, cell cycle and cell differentiation control, immune surveillance, constituting the skeleton of nuclear sub structure, etc. LncRNA plays an important role in individual development and human diseases. This paper mainly reviewed those lncRNAs that have been published, and closely related to eye development and diseases.
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AIM: To compare fungal culture and in vivo confocal microscopy ( IVCM ) in the diagnosis of non- primary fungal keratitis.?METHODS:The clinical data of 31 cases (31 eyes) with non- primary fungaI keratitis from September 2016 to February 2017 in our HospitaI were retrospectiveIy reviewed. The positive rate of the two methods was compared by chi-square test.?RESULTS: The positive rate by fungal culture was 58％(18/31 ) and IVCM was 19％ ( 6/31 ); the positive rate comparison difference was statistically significant between fungal culture and IVCM (x2=7. 56,P<0. 01). In the 13 eyes with positive culture results, 2 eyes were positive by IVCM;in the 25 positive IVCM eyes, 14 eyes were positive in culture.?CONCLUSION: The positive rate of fungal culture in non-primary fungal keratitis is higher than that of IVCM. Fungal culture is an essential auxiliary examination in the diagnosis of non - primary fungal keratitis. With the characteristics of fast, noninvasive and repeatable, IVCM also plays an important role in the diagnosis of non-primary fungal keratitis. The combination of the two methods can improve the positive rate of diagnosis.
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This study looked into a family involving a rare mother-child ABO blood type inconsistency and explored its genetic and molecular basis. In the family, the mother had type AB blood and the father was blood type B and they gave birth to a baby of blood type O. Their blood types were phenotypically identified by using different techniques, including micro-column gel test, immune inhibition test, absorption and elution tests. The sequences of all 7 exons of ABO allele from the core family members were determined by using PCR and clone-based sequencing. The loci of mutated gene were compared against normal human genes. The result showed that the mother's erythrocytes were agglutinable with monoclonal anti-A antibody (2+) and had agglutination reaction with anti-B antibody (4+). The mother's serum registered agglutination action with standard blood type A cells. The findings showed an ABO inconsistency. When domestic antibodies were used, the mother's erythrocytes yielded agglutination reaction with humanized anti-B serum (4+) and anti-B monoclonal antibody but were non-agglutinable with humanized anti-A serum and anti-A monoclonal antibody. Upon absorption and elution, the titer of anit-A antibody was 128 both before and after the absorption test, with no significant difference found between pre- and post-absorption values. Our results confirmed that the mother's allelic gene was type B and contained type A. The father's blood type was type B, and son's blood type was type O. Clone-based sequencing revealed that the mother carried a heterozygous gene of B101.01 (ntA640→G)/O01, which contained an M214→V mutation that could express a weak expression of antigen A, resulting in blood type AB. However, their son did not have the M214→V mutation, which yielded a false ABO-inconsistency between him and his mother. We were led to conclude that type B gene with a M214→V mutation can encode both antigen B and weak antigen B that can lead to false ABO-inconsistencies.
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This study looked into a family involving a rare mother-child ABO blood type inconsistency and explored its genetic and molecular basis. In the family, the mother had type AB blood and the father was blood type B and they gave birth to a baby of blood type O. Their blood types were phenotypically identified by using different techniques, including micro-column gel test, immune inhibition test, absorption and elution tests. The sequences of all 7 exons of ABO allele from the core family members were determined by using PCR and clone-based sequencing. The loci of mutated gene were compared against normal human genes. The result showed that the mother's erythrocytes were agglutinable with monoclonal anti-A antibody (2+) and had agglutination reaction with anti-B antibody (4+). The mother's serum registered agglutination action with standard blood type A cells. The findings showed an ABO inconsistency. When domestic antibodies were used, the mother's erythrocytes yielded agglutination reaction with humanized anti-B serum (4+) and anti-B monoclonal antibody but were non-agglutinable with humanized anti-A serum and anti-A monoclonal antibody. Upon absorption and elution, the titer of anit-A antibody was 128 both before and after the absorption test, with no significant difference found between pre- and post-absorption values. Our results confirmed that the mother's allelic gene was type B and contained type A. The father's blood type was type B, and son's blood type was type O. Clone-based sequencing revealed that the mother carried a heterozygous gene of B101.01 (ntA640→G)/O01, which contained an M214→V mutation that could express a weak expression of antigen A, resulting in blood type AB. However, their son did not have the M214→V mutation, which yielded a false ABO-inconsistency between him and his mother. We were led to conclude that type B gene with a M214→V mutation can encode both antigen B and weak antigen B that can lead to false ABO-inconsistencies.
Subject(s)
Adult , Female , Humans , Pregnancy , ABO Blood-Group System , Genetics , Allergy and Immunology , Base Sequence , DNA Primers , Maternal-Fetal Exchange , Molecular Sequence Data , Mutation , Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology, Nucleic AcidABSTRACT
In order to explore the distribution frequency of endothelial protein C receptor (EPCR) gene A6936G variant and to study the correlation between this mutation and cerebral infarction in Chinese Han population of Hubei province district. The genotype and allele frequencies of EPCR A6936G were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 380 cerebral infarction patients and 380 healthy Chinese Han persons. The results indicated that the frequencies of A/A, A/G and G/G genotypes in cerebral infarction group were 77.1%, 22.1% and 0.8% respectively. While the frequencies of A/A and A/G genotypes in control group were 88.2% and 11.8%, and without G/G genotype. It is concluded that A6936G polymorphism of EPCR can be detected in Chinese Han population of Hubei province district, which may be correlated with the increasing risk of thrombosis in cerebral infarction patients.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antigens, CD , Genetics , Asian People , Genetics , Case-Control Studies , Cerebral Infarction , Genetics , Endothelial Protein C Receptor , Gene Frequency , Genotype , Polymorphism, Genetic , Receptors, Cell Surface , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the inhibitory effect of cyclin-dependent kinase inhibitor p21 on regulation of survivin transcription in human liver cancer HepG cells, and explore the related mechanisms.</p><p><b>METHODS</b>Doxorubicin (DOX) was used to treat HepG cells. Eukaryotic vector pEGFP-C2-p21 was transfected into HepG cells by lipofectamine and positive clones were screened out by G418. The mRNA expression of p21, p53 and survivin were detected by real-time fluorescent quantitative polymerase chain reaction (RQ-PCR). Flow cytometry was used to determine the cell cycle phases, and reverse transcription polymerase chain reaction (RT-PCR) was used to measure the levels of E2F-1 or p300.</p><p><b>RESULTS</b>After treatment with DOX, the expression of p53 and p21 was increased, whereas that of survivin was reduced during 24 hours of the treatment. After transfection the p21 level was 2100.1-fold or 980.9-fold enhanced in comparison with that in HepG2 cells or HepG2-pEGFP cells. Survivin level was markedly down-regulated to 0.5% or 0.6% relative to that in the other two groups, nevertheless, significant p53 changes were not observed. Overexpression of p21 resulted in G1/G0 phase arrest (F = 31.59, P < 0.01), meanwhile, E2F-1 mRNA or p300 mRNA were less expressed compared with that in the other controls (F(E2F-1) = 125.28, P < 0.05; Fp300 = 46.01, P < 0.01).</p><p><b>CONCLUSION</b>p21 could be a potential mediator of survivin suppression at transcription level in HepG2 cells, which might be through the block at G1/G0 phase and down-regulation of transcription factors E2F-1 and p300.</p>
Subject(s)
Humans , Antibiotics, Antineoplastic , Pharmacology , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , Physiology , Doxorubicin , Pharmacology , E2F1 Transcription Factor , Genetics , Metabolism , G1 Phase , Gene Expression Regulation, Neoplastic , Hep G2 Cells , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Metabolism , RNA, Messenger , Metabolism , Resting Phase, Cell Cycle , Transfection , Tumor Suppressor Protein p53 , Genetics , Metabolism , p300-CBP Transcription Factors , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the frequencies of -1516,-574 and 4259 single nucleotide polymorphisms (SNPs) of T cells immunoglobulin mucin -3(TIM-3) gene in Hubei population and address the question whether they are in linkage disequilibrium(LD) .</p><p><b>METHODS</b>Genotypes and allele frequencies of TIM-3 gene were examined by allele-specific polymerase chain reaction (AS-PCR) methods in 147 healthy Hubei Han individuals. Hardy-Weinberg equilibrium and Two-point LD analyses and haplotype frequencies were evaluated with Arlequin v3.1 software.</p><p><b>RESULTS</b>The allele frequencies of the 3 SNPs were in agreement with Hardy-Weinberg equilibrium. Minor allelic frequencies of TIM-3 -1516G/T,-574T/G and 4259G/T were 8.5%,1.0% and 2.0%,respectively. The dominant haplotypes comprising the three loci were G-G-G(2.0%),G-G-T(88.4%), T-G-T(8.5%) and G-T-T(1.0%). LD analyses revealed that all of the coefficient of linkage disequilibrium (D') were 1.</p><p><b>CONCLUSION</b>The -1516,-574 and 4259 loci of TIM-3 gene are in complete linkage disequilibrium. Our study has provided population genetic data on TIM-3 gene in Chinese Hubei Han population and a basis for searching immune-mediated disease-related TIM-3 haplotype.</p>
Subject(s)
Adult , Humans , Male , Alleles , Base Sequence , China , Electrophoresis , Ethnicity , Genetics , Haplotypes , Hepatitis A Virus Cellular Receptor 2 , Linkage Disequilibrium , Genetics , Membrane Proteins , Genetics , Molecular Sequence Data , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Sequence Analysis, DNAABSTRACT
This study was purposed to construct and identify the mammalian expression vector of pEGFP-BMI-1 and to detect whether it could express in human cervix cancer cell line HeLa. The cDNA fragment of BMI-1 obtained by RT-PCR was inserted into pEGFP-N1. The recombinant plasmid was confirmed by restriction enzyme digestion, PCR and DNA sequencing. pEGFP-BMI-1 was transfected into HeLa cells with lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blot analysis. SYBR Green I real-time RT-PCR was used to quantitate P16INK4a mRNA. The results showed that the correct construction of the recombinant plasmid pEGFP-BMI-1 has been shown by restriction enzyme digestion, PCR and DNA sequencing. pEGFP-BMI-1 could express BMI-1-EGFP fusion protein in HeLa cells. Real-time RT-PCR showed that P16INK4a mRNA expression was reduced to 9.2%. It is concluded that the vector of pEGFP-BMI-1 has been successfully constructed and it can be expressed in HeLa cells. This work has laid foundations for further study on biological functions and potential application of BMI-1.
Subject(s)
Humans , Base Sequence , Cyclin-Dependent Kinase Inhibitor p16 , Metabolism , Genetic Vectors , Green Fluorescent Proteins , Genetics , Metabolism , HeLa Cells , Molecular Sequence Data , Nuclear Proteins , Genetics , Metabolism , Polycomb Repressive Complex 1 , Proto-Oncogene Proteins , Genetics , Metabolism , RNA, Messenger , Metabolism , Recombinant Fusion Proteins , Genetics , Metabolism , Repressor Proteins , Genetics , Metabolism , TransfectionABSTRACT
<p><b>AIM</b>To explore a role of G6PD in replenishment of intracellular GSH during oxidative stress.</p><p><b>METHODS</b>In vitro Raji cell was cultured, intracellular GSH levels and G6PD, GR, GPX activities were determined at different time points after PMS treatment when G6PD activity was inhibited or not by DHEA.</p><p><b>RESULTS</b>Intracellular GR, GPX, G6PD activities elevated significantly combined with GSH level decreased dramatically before 30 minutes, replenished gradually after 30 minutes and restore normal levels about 6 h after PMS treatment when G6PD was not inhibited. No change in GR and significant increase in GPX activity were shown following depleted GSH after PMS treatment when G6PD was inhibited by DHEA.</p><p><b>CONCLUSION</b>G6PD contributes to replenish intracellular GSH and is a critical factor regulating GSH levels during oxidative stress.</p>
Subject(s)
Humans , Cell Line, Tumor , Glucosephosphate Dehydrogenase , Metabolism , Glutathione , Metabolism , Glutathione Peroxidase , Metabolism , Oxidation-Reduction , Oxidative Stress , Receptors, Peptide , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the protective effects of hirudin on acute experimental intracerebral hemorrhage (ICH) by observing the changes of histologic pathology and brain water content as well as GFAP-positive cells in the perihematomal brain regions.</p><p><b>METHOD</b>The models of rat ICH were made with infusion of autologous blood into the right neucleus caudatus. The rats were divided randomly into control group, intracerebral hemorrhage group and treating group with hirudin. Brain water content was measured, and pathological and GFAP changes were observed.</p><p><b>RESULT</b>The pathological impairation after ICH were gradually deteriorated and peaked at the third day. Brain water content after ICH was gradually increased and obviously after one day(P < 0.05) and peaked at the third day. GFAP-positive cells were gradually increased and peaked at the seventh day after ICH. In the treating groups, the pathological impairation and brain water content as well as the GFAP-positive cells were decreased as compared to those in the intracerebral hemorrhage group and the control group. And the positive correlation between GFAP-positive cell numbers and brain water content were shown by linear regression.</p><p><b>CONCLUSION</b>The local administration of hirudin, a special inhibitor of thrombin, has protective effects within the first week after ICH.</p>
Subject(s)
Animals , Female , Male , Rats , Acute Disease , Brain , Metabolism , Pathology , Cerebral Hemorrhage , Metabolism , Pathology , Glial Fibrillary Acidic Protein , Metabolism , Hirudins , Pharmacology , Neuroprotective Agents , Pharmacology , Random Allocation , Rats, WistarABSTRACT
Beta-catenin plays a central role in Wnt signaling pathway. The aberrant localization of beta-catenin in nucleus causes the transcription of down-stream target genes, which is the pathogenesy of some solid tumours. As the expression of adheren junction on hemopoietic cells is very low, there are a few studies on beta-catenin expression in leukaemia. This study was aimed to investigate beta-catenin localization and beta-catenin mRNA expression levels in 4 leukemia cell lines so as to explore a new oncogenic mechanism and to find out a new therapeutic target. The beta-catenin localization in leukemia cell lines was detected by immunocytochemistry, the beta-catenin mRNA expression level was assayed by real-time quantitative RT-PCR. The results showed that there was aberrant localization of beta-catenin in Jurkat and Thp-1, and beta-catenin mRNA expression level was not increased in these two cell lines, however, the mRNA expression levels of Jurkat and Thp-1 were lower than those of Daudi and K562. The beta-catenin mRNA expression level was not correlated with beta-catenin aberrant localization in these 4 cell lines. It is concluded that the aberrant localization of beta-catenin may play a role in the development of some leukemia, and the mechanism resulting in beta-catenin aberrant localization not take place at transcription level.
Subject(s)
Humans , Burkitt Lymphoma , Metabolism , Pathology , Cell Line, Tumor , Hematologic Neoplasms , Pathology , Jurkat Cells , K562 Cells , Leukemia, Monocytic, Acute , Pathology , RNA, Messenger , Genetics , Signal Transduction , Wnt Proteins , Metabolism , beta Catenin , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate two polymorphism sites of exon 4 in T cells, immunoglobulin domain and mucin domain protein-1 (TIM-1, also human hepatitis A virus cellular receptor-1) and to detect whether they are associated with allergic asthma in children of the Hans in Hubei province of China.</p><p><b>METHODS</b>The ins/del and IVS 8+9 G/A polymorphisms in TIM-1 were detected with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The genotypes and alleles frequencies were calculated and analyzed.</p><p><b>RESULTS</b>(1) Two alleles, a wide type del and a variant allele ins were identified in the TIM-1 exon 4. The genotype frequencies of ins/ins, ins/del, and del/del were 0.065,0.326, and 0.608 respectively in the healthy population of the Hans. Another IVS 8/9 G/A polymorphism was also found. The genotype frequencies of A/A, G/A, G/G were 0.022, 0.196 and 0.783, respectively. (2) The genotype frequencies of ins/ins, ins/del, and del/del were 0.045, 0.318, and 0.636 respectively in the allergic asthma population in children of the Hans. No significant difference in ins/del polymorphism was found between allergic asthma patients and control subjects. Another 8/9 IVS G/A polymorphism was also found. The genotype frequencies of A/A, G/A and G/G were 0.009, 0.209 and 0.782 respectively in allergic asthma. No significant difference in IVS G/A polymorphism was found between allergic asthma patients and control subjects.</p><p><b>CONCLUSION</b>The genotype and allele frequencies in the two polymorphism sites in TIM-1 in healthy population of the Hans from Hubei province of China were similar to those in Japanese. The two polymorphism sites of TIM-1 are not associated with allergic asthma in Chinese children.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Asthma , Genetics , Cell Cycle Proteins , Genetics , China , Gene Frequency , Genotype , Intracellular Signaling Peptides and Proteins , Genetics , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment LengthABSTRACT
Objective To investigate the relationship between the exon 4 gene polymorphism of TIM-1 and rheumatoid arthritis (RA) in Han population from Hubei province.Methods Polymerase chain reaction was used to detect the ins/del polymorphism of the exon 4 of TIM-1 from RA population and the normal con- trols.Rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP) antibody and anti-keratin antibody (AKA) were also detected.Results Two alleles,a wild type del and a variant allele ins were identified in the TIM-1 exon 4.The genotype frequencies of del/del,ins/del and ins/ins were 0.650,0.280 and 0.070 respec- tively in the normal controls and 0.616,0.302,0.082 respectively in RA population.There was a significant correlation between the positive ratio of AKA and the genotypes of the exon 4.Conclusion The polymorphism of the exon 4 of TIM-1 is not associated with rheumatoid arthritis in Han population from Hubei Province of China.The genotypes of the exon 4 may have an effect on the expression of AKA.
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<p><b>OBJECTIVE</b>To determine whether 2 polymorphism sites of the high-affinity IgE receptor beta (Fc epsilon RI beta) gene were associated with allergic asthma in Han nationality of Hubei province in China.</p><p><b>METHODS</b>DNA and clinical data were obtained from allergic asthma patients and were compared with those from a group of healthy control subjects. The subjects were genotyped for the -109C/T and a coding variant Glu237Gly in Fc epsilon RI beta gene by polymerase chain reaction-restriction fragment length polymorphism.</p><p><b>RESULTS</b>(1) The genotype frequencies were 0.403 for -109T/T, 0.491 for -109T/C and 0.106 for -109C/C in allergic asthma patients in a Chinese population. No significant difference in the distribution of -109C/T polymorphism was found between allergic asthma patients and healthy control subjects; however, a homozygosity for the -109T allele was associated with increased total plasma IgE levels in patients with allergic asthma (F=7.523, P<0.01). (2) The frequency of Gly237Gly genotype was 0.051 in patient group, and 0.020 in control group. The allele frequencies of Gly237 in the patients and control were 0.236 and 0.136 respectively. There was a significant association between Gly/Gly genotype and allergic asthma. Among allergic asthma patients Gly237Gly was significantly associated with high IgE.</p><p><b>CONCLUSION</b>These data suggested that the Gly237Gly genotype of the Fc epsilon RI beta gene conferred genetic susceptibility to allergic asthma in Chinese, which affected the total plasma IgE levels in the allergic asthma patients. And a homozygosity for the -109T allele was associated with increased total plasma IgE.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Asthma , Blood , Genetics , China , Gene Frequency , Genetic Predisposition to Disease , Genetics , Genotype , Immunoglobulin E , Blood , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length , Promoter Regions, Genetic , Genetics , Receptors, IgE , GeneticsABSTRACT
Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.