Objective The purpose of this paper is to study PGM1 genotyping by PCR RFLP.Method 300 unrelated individuals of Han were genotyped using PCR RFLP. The target amplificaton products of extron 4 and 8 of PGM1 gene were digested by Bgl II and Nla III respectively.The digested DNA fragments were typed by PAGE.Result This PGM1 RFLP system can discriminate 9 genotypes with Dp of 0 7450 in Han population.Compared with conventional PAGIEF, 1+2- and 1-2+ cant be differentiated and the rare genotypes also cant be detected by this method.The advantage of this method was PGM1 genotyping successfully in bloodstains stored for 25 years and with 0 1ng genomic DNA.PGM1 RFLP method is useful for forensic identification.

Since the information supplied by the paternity testing of alleged parents was less than that of standard triplet parentage testing,so the paternity index (PI) calculating methods of standard triplet parentage testing was not suitable for calculating the PI value of alleged parents.In order to establish a more precise method for calculating PI value of alleged parents with STR typing results,the first thing is to summarize the standard triplet PI calculating formulas according to the Essen Mller theory.These formulas are 1/p,1/2p,1/p+q,1/2p+2q.This article reports a new PI calculating method in case of paternity testing of alleged parents.Compared with other methods,the new method for calculating Y value either considering random man and random female or considering the alleged father(mother)and random female(man).

DNA methylation is a post-replication modification that is predominantly found in cytosines of the dinucleotide sequence CpG. Epigenetic information is stored in the distribution of the modified base 5-methylcytosine. DNA methylation profiles represent a more chemically and biologically stable source of molecular diagnostic information than RNA or most proteins. Recent advances attest to the great promise of DNA methylation markers as powerful future tools in the clinic. In the past decade, DNA methylation analysis has been revolutionized by two technological advances--bisulphite modification of DNA and methylation-specific polymerase chain reaction (MSP). The methylation pattern of human genome is space-time specific, sex-specific, parent-of-origin specific and disease specific, providing us an alternative way to solve forensic problems.
Base Sequence ; CpG Islands/genetics* ; DNA/blood* ; DNA Fingerprinting/methods* ; DNA Methylation ; Epigenesis, Genetic ; Forensic Medicine/methods* ; Genetic Markers ; Genome, Human ; Humans ; Paternity ; Polymerase Chain Reaction/methods*

The prognostic value of phosphatidylinositol-4,5-bisphosphate 3-kinase,catalytic subunit alpha (PIK3CA) in patients with esophageal squamous cell carcinoma (ESCC) is controversial.We aimed to investigate the prognostic significance of PIK3CA mutation in patients with ESCC.EMBASE,PubMed,and Web of Science databases were systematically searched from inception through Oct.3,2016.The hazard ratios (HRs) and 95％ confidence intervals (CI) were calculated using a random effects model for overall survival (OS) and disease-free survival (DFS).Seven studies enrolling 1505 patients were eligible for inclusion of the current meta-analysis.Results revealed that PIK3CA mutation was not significantly associated with OS (HR:0.90,95％ CI:0.63-1.30,P=0.591),with a significant heterogeneity (I2=65.7％,P=0.012).Additionally,subgroup analyses were further conducted according to various variables,such as types of specimen,the sample size,technique and statistical methodology.All results suggested that no significant relationship was found between PIK3CA mutation and OS in patients with ESCC.For DFS,there was no significant association between PIK3CA mutation and DFS in patients with ESCC (HR:1.00,95％ CI=0.47-2.11,P=0.993,I2=73.7％).Publication bias was not present and the results of sensitivity analysis were very stable in the current meta-analysis.Our findings suggest that PIK3CA mutation has no significant effects on OS and DFS in ESCC patients.More well-designed prospective studies with better methodology for PIK3CA assessment are required to clarify the prognostic significance of PIK3CA mutation in ESCC patients.

OBJECTIVE: The genetic polymorphism of two STR loci, D20S85 and D6S477, were studied in 280 unrelated Chinese individuals in Wuhan. METHODS: The PCR amplified products were analyzed by PAGE and silver staining. RESULTS: 10 and 9 alleles were observed in these two STR loci, and the discriminating power (DP) were 0.9085 and 0.9127 respectively. No deviations from Hardy-Weinberg equilibrium were found. The two STR loci had been successfully applied to individual identification and paternity testing. CONCLUSION The results demonstrated that the two loci were useful for forensic identification.
Alleles ; Asian People/genetics* ; China ; Forensic Medicine ; Gene Frequency ; Humans ; Polymorphism, Genetic ; Tandem Repeat Sequences/genetics*

OBJECTIVETo investigate the significance of clinicopathological stage of chronic idiopathic myelofibrosis (CIMF) in WHO classification of 2001.

METHODSHistopathological analysis of bone marrow biopsy plastic-embedded sections stained with H-G-E and Gomori's stains and clinical features of 113 cases previously diagnosed as primary myelofibrosis (PMF) and 48 cases MPD-U (total of 161 cases which including male 79 and female 82) were studied retrospectively.

RESULTSThere was no significant differences on the clinical features among the cellular phase, collagen fiber phase, sclerotic phase and osteomyelosclerosis of 113 previously diagnosed patients. According to WHO classification 2001 of CIMF, previously diagnosis in 48 cases with MPD-U was WHO pre-CIMF, and in 113 cases with PMF was WHO CIMF-Fs. There were significant differences between of WHO pre-CIMF and WHO CIMF-Fs about clinicopathological features except age. The percentage of immature granulocytes, normoblasts, lymphocytes in peripheral blood, the size of hepatosplenomegaly, and the percent age of tear drop-like red blood cells in pre-CIMF were significantly lower than those in CIMF-Fs (P < 0.05). However, the number of hemoglobin and platelets in patients with pre-CIMF were significantly higher than that with CIMF-Fs (P < 0.01).

CONCLUSIONpre-CIMF and CIMF-Fs in clinical and histopathological features were different development stage of CIMF, while osteomyelosclerosis is a variant of CIMF, but not an independent disease.

Adult ; Aged ; Biopsy ; Bone Marrow ; pathology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Primary Myelofibrosis ; classification ; pathology ; Thrombopoiesis

It was expected that recombinant Aspergillus niger glucose oxidase could be expressed in Trichoderma reesei with stable activity. T. reesei CBHI promoter--CBHI ss. gene--A. niger glucose oxidase gene--T. reesei CBHI terminator--A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator--pUC19 (pCBHGOD) vector was constructed in E. coli DH5alpha by PCR application and gene cloning methods. T. reesei QM9414 protoplast was transformed by T. reesei CBHI promoter-CBHI ss. Gene--A. niger glucose oxidase gene--T. reesei CBHI terminator-A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator linear DNA fragment (CBHGOD fragment) that was made by digestion of pCBHGOD with Kpn I. T. reesei mutant clone with homologous recombinant A. niger glucose oxidase gene was selected by PCR method. Recombinant glucose oxidase was produced by mutant T. reesei strain under induction of wheat straw for 5 days. Recombinant glucose oxidase molecular mass was showed the same as native A. niger glucose oxidase standard from Sigma company by Western blot analysis. Recombinant glucose oxidase activity was 25u/mL in medium. The yield was 0.5 g/L in comparison with Sigma company glucose oxidase standard. There was no recombinant GOD degradation during Trichoderma reesei cultivation that was showed in Western blot analysis. Trichoderma reesei has capability to be a new recombinant host for Aspergillus niger GOD production.
Aspergillus niger ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Glucose Oxidase ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Trichoderma ; genetics ; metabolism

BACKGROUNDMultiple myeloma (MM) is a malignant tumor, which takes the second place in malignant blood disease. The clinical symptoms are complicated that make more difficult to diagnose and therapy. Lots of researches focus on the proteins about MM in order to solve those problems. We used proteomic methods to find potential biomarkers in MM patients.

METHODSWe applied the peptide ligand library beads (PLLBs) to deplete high abundance proteins in serum for finding potential pathogenic factors and biomarkers of MM. Using 1D-Gel-liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified 789 and 849 unique serum proteins in MM patients and in healthy controls, respectively.

RESULTSTwenty-two proteins were found differentially expressed between the two groups including serum amyloid A protein, vitamin D-binding protein isoform-1 precursor, plasma kallikrein, and apolipoprotein A-I. Changes of integrin alpha-11 and isoform-1 of multimerin-1 were validated with Western blotting. The linkage of the differentially expressed proteins and the pathogenesis pathways of MM were discussed.

CONCLUSIONSPLLB combined with 1D-gel-LC-MS/MS analysis is an efficient method to identify differentially expressed proteins in serum from patients with MM.

Biomarkers ; blood ; Biomarkers, Tumor ; blood ; Humans ; Multiple Myeloma ; blood ; Peptide Library ; Proteomics ; methods ; Tandem Mass Spectrometry

OBJECTIVETo establish a novel method for the multiplex analysis of the methylation and single nucleotide polymorphism (SNP).

METHODSThe imprinted SNP rs220028 was chosen as a model. Genomic DNA, after being digested with methylation sensitive restriction enzyme, were typed by mutagenically separated PCR (MS-PCR). The polymorphism of restriction site was excluded by PCR-RFLP.

RESULTSBy post-digestion MS-PCR, the methylated allele was detected selectively, the maternal origin of which was confirmed by pedigree analysis; A=0.5085, G=0.4915,PIC=0.3749.

CONCLUSIONThe multiplex analysis of methylation markers and SNP can be achieved by post-digestion MS-PCR. The imprinted SNP locus rs220028 is a potentially useful marker in screening Prader-Willi/Angelman syndrome.

DNA Methylation ; DNA Restriction Enzymes ; metabolism ; Genetic Markers ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the early changes of the blue-on-yellow (B/Y) perimetry in patients with early primary open-angle glaucoma(POAG).

METHODSThirty-one cases (45 eyes) of POAG underwent the central 30 degrees field examination of B/Y as well as routine white-on-white (W/W).

RESULTNo significant difference of mean index deficiency (MD) between B/Y and W/W perimetry was detected (P>0.05). However, there were marked changes in index GHT (glaucoma hemisphere test) and more defect points in B/Y visual field than those in W/W (P<0.01).

CONCLUSIONB/Y perimetry might be more sensitive than W/W perimetry a potentially more and valuable method in detection of early POAG lesion.

Adult ; Female ; Glaucoma, Open-Angle ; diagnosis ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Visual Field Tests ; Visual Fields