Correlation between JAK2-V617F mutations and variation of peripheral blood cells among BCR/ABL-negative MPD patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the correlation between JAK2-V617F mutation and complete blood counts among patients with BCR/ABL-negative myeloproliferative diseases (MPD).</p><p><b>METHODS</b>One hundred and ninety one patients were recruited. Retrospectively, their laboratory data were analyzed for the counts of red blood cells (RBC), white blood cells (WBC) and platelets (PLT). And the incidence of JAK2-V617F mutation was determined.</p><p><b>RESULTS</b>There was significant difference in the incidence of JAK2-V617F mutation between patients with different cell counts (P< 0.01). The incidence of JAK2-V617F mutation has increased with the counts of RBC and PLT, which was the highest (92.86%) among those featuring simultaneous increase in all three series.</p><p><b>CONCLUSION</b>The incidence of JAK2-V617F mutation seems to be strongly associated with variation of peripheral blood cell counts among patients with BCR/ABL-negative MPD. Variation of peripheral blood cells, particularly RBC, may be correlated with the rate of JAK2-V617F mutation.</p>

Detection of epidermal growth factor receptor gene mutation in non-small cell lung cancer by allele-specific oligonucleotide-PCR and bi-loop probe specific primer quantitative PCR / 中华病理学杂志

<p><b>OBJECTIVE</b>To compare the detection sensitivity of epidermal growth factor receptor (EGFR) mutations between allele specific oligonucleotide PCR (ASO-PCR) and bi-loop probe and specific primer quantitative PCR (BPSP-qPCR).</p><p><b>METHODS</b>A total of 96 non-small cell lung cancer specimens were selected from West China Hospital from September 2009 to December 2010. ASO-PCR was developed to detect the presence of classical EGFR mutations. A total 39 available specimens were also tested by BPSP-qPCR.</p><p><b>RESULTS</b>EGFR mutation detection rate was 30.2% (26/96) by ASO-PCR. The mutation rate was higher in female than in male patients [45.5% (20/44) vs. 17.3% (9/52), P = 0.003], non-smokers than smokers [44.1% (26/59) vs. 8.1% (3/37), P < 0.001] and adenocarcinomas than other subtypes of lung cancer [37.0% (27/73) vs. 8.7% (2/23), P = 0.01]. Among mutation negative cases by ASO-PCR, BPSP-qPCR increased the rate of detection of 19-del and L858R mutation by 10.3% (4/39) in adenocarcinomas and non-smoking subset. Overall, the mutation detection rate of BPSP-qPCR was higher than that of ASO-PCR [66.7% (26/39) vs. 41.0% (16/39), P = 0.02].</p><p><b>CONCLUSION</b>BPSP-qPCR has a better detection sensitivity than that of ASO-PCR.</p>

GenoType MTBDRplus assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis in Sichuan / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the molecular and epidemic characteristics of rifampin (RFP) and isoniazid (INH) resistance of mycobacterium tuberculosis (MTB) in Sichuan.</p><p><b>METHODS</b>GenoType reg; MTBDRplus Assay GTplus was used to examine 68 clinical isolates of MTB and 105 clinical specimens for mutations in rpoB, katG and inhA genes related to RFP and INH resistance.</p><p><b>RESULTS</b>Of the 151 valid tests obtained, 44 (29.14%) and 26 (17.22%) showed drug resistance and multidrug resistance, respectively. Resistance to RFP and INH was found in 21.85% (33/151) and 24.50% (37/151) of the samples, respectively. The most prevalent mutations were rpoB S531L, katG S315T1 and inhA C-15T. The multidrug resistance rate in the sputum specimens was significantly higher than that in the non-respiratory samples (19.35% vs 7.41%).</p><p><b>CONCLUSION</b>Drug-resistant, especially multidrug-resistant tuberculosis is highly prevalent in Sichuan. The multidrug-resistant bacteria most frequently show rpoB S531L combined with katG S315T1 mutations, suggesting the necessity of developing rapid clinical identification methods for drug-resistant MTB to control the spread of the resistant strains.</p>

Detection of epidermal growth factor receptor gene mutations in non-small cell lung cancer using bi-loop probe specific primer quantitative PCR / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the sensitivity of bi-loop probe and specific primer quantitative PCR (BPSP-qPCR) in the detection of epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>BPSP-qPCR was employed to examine the presence of mutations of EFGR exon 19 through 21. Correlation of the mutations with clinicopathological characteristics and types of tumor samples were performed.</p><p><b>RESULTS</b>In the cohort of 265 specimens, 30.2% (80/265) mutations were found to be 19-del and/or L858R. Females (39.7%, 31/78), non-smokers (41.0%, 43/105) and adenocarcinoma patients (37.8%, 51/135) had a higher mutation rate (P<0.05) among 184 patients whose profiles were available. T790M combined with 19-del and/or L858R accounted for 3.3% (6/184) of the mutations. Male metastatic tumors (29.6%, 8/27), pleural fluids of females (42.9%, 9/21) and non-smokers (40.7%, 11/27) were found to have higher percentage of 19-del and/or L858R mutations, in contrast, no mutations were found in the metastatic lesions of non-adenocarcinoma patients (P>0.05).</p><p><b>CONCLUSIONS</b>BPSP-qPCR is a robust method in detection of EGFR mutations with high consistency and sensitivity. The difference of EGFR mutations in primary tumors, metastatic lesions and pleural fluids suggests that EGFR tyrosine kinase inhibitors (EGFR-TKI) treatment may have variable treatment effects depending on the tumor sites.</p>

Effects of single nucleotide polymorphisms 869 T/C and 915 G/C in the exon 1 locus of transforming growth factor-beta1 gene on chronic obstructive pulmonary disease susceptibility in Chinese / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The main risk factor for chronic obstructive pulmonary disease (COPD) is cigarette smoking. However, only 10% - 20% of chronic heavy smokers develop systematic COPD. We hypothesized that the inheritance of gene polymorphisms could influence the development of COPD, which was investigated by studying two single nucleotide polymorphisms (SNP) in exon 1 of the transforming growth factor-beta1 (TGF-beta1) gene.</p><p><b>METHODS</b>We enrolled 219 patients with COPD as the research group and 148 healthy people as the control group, all of whom were Chinese Han people. The polymorphisms of the TGF-beta1 gene, 869T/C and 915G/C, were analyzed using the method of amplification refractory mutation system-polymerase chain reaction (ARMS-PCR).</p><p><b>RESULTS</b>The occurrence of the TGF-beta1 gene 869T/C polymorphism in patients with COPD was significantly different from the control group (P < 0.05), in which the relative risk of this disease increased in cases who had the C allele (OR: 1.131, 95%CI: 1.101 - 1.539). There was no increased frequency of TGF-beta1 915G/C gene in COPD patients compared with control subjects (P > 0.05).</p><p><b>CONCLUSIONS</b>The polymorphism 869T/C in TGF-beta1 gene has a significant association with disease occurrence in COPD patients and the C allele might be a risk factor. The homozygous wild-type CC of 869T/C on TGFbeta1 could be a predisposing factor in COPD and those who carry the C allele might have particularly susceptibility to developing COPD.</p>

Analysis of the genetic polymorphism of 17 Y-chromosomal short tandem repeat loci in the Han population in Chengdu / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain the population genetic data of 17 Y-chromosomal short tandem repeat (Y-STR) in the Han population in Chengdu of Sichuan Province.</p><p><b>METHODS</b>The 17 Y-STR loci were amplified from the blood samples of 111 unrelated Chengdu Han individuals using the AmpFlSTR Yfiler system. The PCR products were genotyped with an ABI 3130 genetic analyzer.</p><p><b>RESULTS</b>In the loci of in DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385a/b, DYS393, DYS391, DYS439, DYS635, DYS392, Y-GATA-H4, DYS437, DYS438, and DYS448, 3 to 8 alleles were detected in the Han population in Chengdu, and 36 alleles were detected in the locus DYS385a/b, with the minimal gene diversity (GD) value of 0.3970 (DYS391) and maximal value of 0.9561 (DYS385a/b). The DNA samples of 16 women and 7 different species of animals were amplified, but no specific products were found for the 17 Y-STR loci. No mutations of the 17 Y-STR alleles were observed in 20 father-son pairs as confirmed by autosomal STR analysis.</p><p><b>CONCLUSION</b>The 17 Y-STR loci are highly polymorphic and are suitable for personal identification, paternity testing, population genetics and anthropology studies.</p>

Association of the Pro12Ala polymorphism in peroxisome proliferators activated receptor-gamma gene with rheumatoid arthritis in Sichuan Province of China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of the Pro12Ala variant in peroxisome proliferators-activated receptor gamma (PPAR gamma) gene with rheumatoid arthritis.</p><p><b>METHODS</b>The genotypes of the Pro12Ala variant in the PPAR gamma gene were determined by polymerase chain reaction-restriction fragment length polymorphism in 421 unrelated subjects of the Han population in the Sichuan Province of China, including 207 subjects with rheumatoid arthritis and 214 subjects without the disease. The clinical data were also collected and analyzed.</p><p><b>RESULTS</b>The allele frequencies in the case and control groups were 98.79%, 95.79% for allele P and 1.21%, 4.21% for allele A; the genotype frequencies were 97.58% and 91.59% for PP, 2.42% and 8.41% for PA, and 0 for AA. The A allele frequency was much lower in the RA group than that in the control group.</p><p><b>CONCLUSION</b>The above data showed that the Pro12Ala variant of the PPAR gamma was associated with rheumatoid arthritis. The A allele might be a protective factor for RA. The Pro12Ala polymorphism in the PPAR gamma gene in Sichuan Han population is similar to that in other populations in China, but different from that in European and American populations.</p>

Identification of sarcosaphagous flies by analyses the sequence of 16S rDNA / 法医学杂志

OBJECTIVE@#To solve the difficulties of identification of Sarcosaphagous flies such as Lucilia sericata (Meigen) and Lucilia cuprina (Wiedemann) which could not be identified by analyzing the 278bp and 635 bp regions of the gene encoding for cytochrome oxidase subunit I and II (CO I and CO II) in mtDNA.@*METHODS@#Specimens were collected from the corpses of rabbits on the grassland in Huhhot and Chengdu, the sequences of 551 bp region of 16S rDNA of their mtDNA were analyzed, the multiple-alignment program DNAMAN(version 4.0) and MEGA 2.1 sofeware were employed for sequence alignments neighbour-joining tree construction.@*RESULTS@#Lucilia sericata (Meigen) and Lucilia cuprina (Wiedemann) were distinguished successfully by sequence analysis of The 551 bp region of the gene of 16S rDNA.@*CONCLUSION@#The 551 bp region of the gene of 16S rDNA of sarcosaphagous flies can be used for identifying them on species level effectively. It is likely to be a successful compliment to identify the sarcosaphagous flies by sequence analysis of CO I and CO II in mtDNA.

Distribution of haplotypes for four Y-sTR loci and validation in forensic science by using a double-fluorescent multiplex PCR system / 法医学杂志

OBJECTIVE@#We focus on developing a multiplex PCR system for Y-STR loci that can be detected by double fluorescent system and assessing their usefulness in forensic mixture samples.@*METHODS@#The primers of four Y-STR loci (DYS-GATA-A10, DYS531, DYS557 and DYS448) amplified by multiplex PCR technique were labeled with fluorescence, then the PCR products of these Y-STRs loci were detecting and typing by ABI PRISM310 Genetic Analyzer.@*RESULTS@#When 120 unrelated individuals from the Han population in Chengdu were detected by the system, Y-GATA-A10, DYS531, DYS557 and DYS448 showed 5, 5, 8, 7 alleles, respectively. A total of 78 different haplotypes was identified and the genetic diversity reached 0.9881. To the three cases of mixture stains failed by using conventional autosomal STR analysis, our multiplex system drew conforming conclusion comparing to the suspect's Y-STRs genotypes.@*CONCLUSION@#Our results show that the multiplex system of four Y-STR will be very powerful for Y-STR database establishing, the paternity testing and mixture stains identifying.

The study of the sequence of molecular markers of mitochondrial DNA of Sarcosaphagous Flies / 法医学杂志

Identifying sarcosaphagous flies specimens is an important first step in a forensic-entomological analysis. It is traditionally performed using morphological features of the Sarcosaphagous Flies. However, Morphological identification may be complicated by the numerical diversity of species and physical similarity between different species, particularly in immature stages. The sequences focused on some sections of the cytochrome oxidase I and II (CO I and CO II) encoding region of mtDNA could be as the prospective basis of a diagnostic technique. By Analysis of these sequences, forensic doctors can reveal abundant phylogenetic informative nucleotide substitutions that could effectively identify Sarcosaphagous flies to species group. It was not reported in our country before and was reviewed in this article now.

Distributions of haplotypes for three Y-STR loci in a Tibetan ethnic group of Chinese population by using Y-STR multiplexes / 法医学杂志

OBJECTIVE@#One multiplex genotyping system was developed in using silver staining with allelic ladders for three Y-chromosome STR markers (DYS434, DYS443, and DYS456), with a view towards the application of rapid and simple genotyping assay methods for DNA profiling. The distributions of haplotypes for three Y-STR loci(DYS434, DYS443, and DYS456) was investigated in a Tibetan ethnic group of Chinese population.@*METHODS@#Allele and haplotype frequencies at these Y-STRs loci(DYS434, DYS443, and DYS456) were analysed by PCR amplification using Y-STR multiplexes, followed by horizontal non-denaturing polyacrylamide gelelec-trophoresis in 101 unrelated males of Tibetan ethnic group in Lasa of China.@*RESULTS@#A total of 31 different haplotypes were found, 16 of them being unique. The haplotype diversity value (which is the same as the discrimination index) calculated from all three loci combined was 0.9481, which is informative.@*CONCLUSION@#The Y-STR multiplexes provide useful information for forensic analysis and paternity tests and can also be of great benefit for providing information not normally available from autosomal DNA systems.