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28 August 2017, Volume 33 Issue 4 Previous Issue    Next Issue

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Gene Analysis for the Sudden Death of Hypertrophic Cardiomyopathy by Whole Exome Sequencing XU CHUAN-CHAO, BAI YUN-ZHI, XU XIN-SHU, ET AL. 2017, 33(4): 339-343.  DOI: 10.3969/j.issn.1004-5619.2017.04.001 Abstract ( 663 )   Objective To analyze the related pathogenicity gene mutations in a sudden death of hypertrophic cardiomyopathy （HCM） on whole exome level. Methods Whole exome sequencing （WES） was been performed on a sudden death case sample with pathological features of HCM by Illumina?誖Hiseq 2500 platform. Using hg19 as the reference sequences, the sequencing data were analyzed. Suspicious single nucleotide variants （SNV） were screened, and the conservatism and function were analyzed by the software such as PhyloP, PolyPhen-2, SIFT, etc. Results After screening, a heterozygous mutation C719R was finally identified in the gene MYBPC3 of this case. Conclusion The molecular anatomy on whole exome level by second generation sequencing technology can help to define the molecular mechanism of HCM and provide a new mothed and thought for analysis of death cause. Related Articles | Metrics

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Relationship between Wound Age and the Expression of Fzd2 in Rats Skeletal Muscle after Contusion DONG TA-NA, PEI MING, LI NA, ET AL. 2017, 33(4): 344-347.  DOI: 10.3969/j.issn.1004-5619.2017.04.002 Abstract ( 527 )    Objective To investigate the relationship between wound age and the expressions of frizzled-2 （Fzd2） mRNA and its protein in rats skeletal muscle after contusion, and to explore its possibility of being an index for wound age estimation. Methods The mRNA and protein expressions of Fzd2 in rats skeletal muscle of the control group and the experimental group within 4-48 h after contusion were detected per 4 h by RT-qPCR and Western blotting. Results The relative expression of Fzd2 mRNA increased at 24 h, 36 h and 40 h after contusion, and the expression at 24 h was twice as the control group （P<0.05）. The relative expression of Fzd2 protein changed inconspicuously after contusion （P>0.05）. Conclusion The changes of Fzd2 mRNA expression after contusion in a certain time can be a basis to estimate wound age by combined multi-indicators. Related Articles | Metrics

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Screening of Differential Expression Proteins in Rat Brain Tissues after DAI by iTRAQ-LC-MS/MS CHEN QING, BAI JIE, ZHANG WEN-FANG 2017, 33(4): 348-352.  DOI: 10.3969/j.issn.1004-5619.2017.04.003 Abstract ( 691 )   Objective To screen for the differential expression proteins in brain tissues of SD rat after diffuse axonal injury （DAI） by isobaric tag for relative and absolute quantification-liquid chromatograph-mass spectrometer/mass spectrometer （iTRAQ-LC-MS/MS）, and to explore potential biomarkers available for the diagnosis of DAI. Methods Animal models of DAI were established with the Marmarou method as reference, and the subjects were divided into blank control group （n=4）, sham strike group （n=4） and fatal strike group （n=4）, respectively. The proteins in rat brain tissues were detected by iTRAQ-LC-MS/MS, and bioinformatics analysis and verification were performed on the results and screened for the differential expression proteins. Results A total of 2 016 proteins were identified and quantified. The bioinformatics analysis revealed that the proteins had wide distribution and function, and participated in different biological processes. There were 16 proteins showed differential expression in fatal strike group, including one up-regulated expression protein and 15 down-regulated expression proteins. The results of iTRAQ-LC-MS/MS were confirmed by Western blotting method. Conclusion Multiple differential expression proteins in rat brain tissues after DAI can be screened by iTRAQ-LC-MS/MS. This not only indicates a research direction for exploring the pathogenesis of DAI, but also provides potential biomarkers available for the diagnosis of DAI. Related Articles | Metrics

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Comparison of Measurement Methods of Exophthalmos and Its Forensic Significance TAN SI-LEI, CHEN JIE-MIN, YU XIAO-YING, ET AL. 2017, 33(4): 353-356.  DOI: 10.3969/j.issn.1004-5619.2017.04.004 Abstract ( 897 )    Objective To compare the differences between Hertel exophthalmometry and CT on exophthalmos measurement, and explore its practical application value in forensic medicine. Methods Fifty-six normal individuals without eye injuries or diseases were selected as the control group. The absolute values of exophthalmos were measured in the standardized CT image workstation. The difference of binocular exophthalmos was compared in normal group. Forty-seven cases with unilateral orbital fracture were divided into injured eye group and normal eye group according to the presence of binocular orbital fractures. The differences of the absolute values of eophthalmos from the same eye and the relative values of eophthalmos between two eyes for same person measured by Hertel exophthalmometry and CT were analyzed. Results There was no statistical difference of exophthalmos between the normal eyes measured by CT method. In the normal eye group, the absolute value of exophthalmos measured by CT method was （16.66±5.41） mm, which was （16.16±4.45） mm when measured by the Hertel exophthalmometry method. There was no statistical difference between two groups （P>0.05）. In the injured eye group, there was statistical difference between the absolute values of exophthalmos measured by two measurement methods （P<0.05）. There was no statistical difference of the relative values of exophthalmos between the injured eyes and the normal eyes by two measurement methods （P>0.05）. Conclusion CT method has a good consistency with Hertel exophthalmometry, which can be applied into the practice of medicolegal expertise. Related Articles | Metrics

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Identification of Y-chromosomal Genetic Types for the Soldier’s Remains from Huaihai Campaign WANG CHI-ZAO, WEN SHAO-QING, SHI MEI-SEN, ET AL. 2017, 33(4): 357-362.  DOI: 10.3969/j.issn.1004-5619.2017.04.005 Abstract ( 788 )   Objective To identify the Y-chromosomal genetic types for the soldier’s remains from Huaihai Campaign, and to offer a clue for search of their paternal relatives. Methods DNA of the remains were extracted by the ancient DNA extraction method. Yfiler kit was used for the multiplex amplification of 17 Y-STR loci. The haplogroups of the samples were speculated. Detailed genotyping of the selected Y-SNP was performed based on the latest Y-chromosome phylogenetic tree. Haplotype-sharing analysis was done based on the data of Y-SNP and Y-STR, the closest modern individual information to the genetic relationship of remains was gained. Results A total of 8 Y-STR haplotypes were observed on 17 Y-STR loci of 8 male individuals. Furthermore, 6 Y-SNP haplogroups were identified, which were O2a1-M95+, O1a1-P203+, O3*-M122+/M234-, D1-M15+, C3*-ST and R1a1-M17+. Conclusion Identification of Y-chromosomal genetic types for the soldier’s remains from Huaihai Campaign shows a reference value on inferring the geographical origins of old materials. Related Articles | Metrics

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Formula Derivation and Validation of Probability of Exclusion in the Cases of Standard Triplet Parentage Testing ZHOU MI, ZHANG HAN-QIU, WANG JUN 2017, 33(4): 363-367.  DOI: 10.3969/j.issn.1004-5619.2017.04.006 Abstract ( 1406 )   Objective To derive and experiment validate the probability of exclusion （PE） formulas in the cases of standard triplet parentage testing. Methods The formulas were derived voluntarily based on the PE definition: PE=■Pi2（1-Pi）2+■PiPj（1-Pi）3+■PiPj（1-Pj）3+■PiPj（Pi+Pj）（1-Pi-Pj）2. This formula was compared with the 5 formulas （1）-（5） reported previously, and the PE values of 19 autosomal STR loci in AGCU EX20 system were calculated. Based on 1 000 samples of single-parentage cases and 1 000 unrelated individuals, the real experiment was designed and the real experiment results of PE were calculated. Ten million pairs of simulated biological mother and son and 10 million random individuals were gained by random simulation method, and the simulated experiment was designed and the simulated values of PE were calculated. In 19 STR loci, the sum of all allele frequency （S） was calculated, and the formula values of PE were compared with the values of real and simulated experiments. Results If S=1, the calculation values of formula （1）, （2）, （5） and （6） were quite the same, which accord with the double verification of real and simulated experiments. If S≠1, there was a minor error in the calculation results of formula （1）, （2）, （5） and （6）, while which had a large error in formula （3） and （4）. Conclusion The formula （6） derived in present study and the classical formulas （1）, （2） and （5） can be applied to the standard triplet parentage testing. The S value has a certain influence on PE calculation. Related Articles | Metrics

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Whole Genome Sequencing of Human mtDNA Based on Ion Torrent PGMTM Platform CAO YU, ZOU KAI-南, HUANG JIANG-PING, ET AL. 2017, 33(4): 368-373.  DOI: 10.3969/j.issn.1004-5619.2017.04.007 Abstract ( 527 )   Objective To analyze and detect the whole genome sequence of human mitochondrial DNA （mtDNA） by Ion Torrent PGMTM platform and to study the differences of mtDNA sequence in different tissues. Methods Samples were collected from 6 unrelated individuals by forensic postmortem examination, including chest blood, hair, costicartilage, nail, skeletal muscle and oral epithelium. Amplification of whole genome sequence of mtDNA was performed by 4 pairs of primer. Libraries were constructed with Ion ShearTM Plus Reagents kit and Ion Plus Fragment Library kit. Whole genome sequencing of mtDNA was performed using Ion Torrent PGMTM platform. Sanger sequencing was used to determine the heteroplasmy positions and the mutation positions on HVⅠ region. Results The whole genome sequence of mtDNA from all samples were amplified successfully. Six unrelated individuals belonged to 6 different haplotypes. Different tissues in one individual had heteroplasmy difference. The heteroplasmy positions and the mutation positions on HVⅠ region were verified by Sanger sequencing. After a consistency check by the Kappa method, it was found that the results of mtDNA sequence had a high consistency in different tissues. Conclusion The testing method used in present study for sequencing the whole genome sequence of human mtDNA can detect the heteroplasmy difference in different tissues, which have good consistency. The results provide guidance for the further applications of mtDNA in forensic science. Related Articles | Metrics

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Forensic Analysis for 54 Cases of Suxamethonium Chloride Poisoning ZHAO YUAN-FENG, ZHAO BING-QING, MA KE-JING, ET AL. 2017, 33(4): 374-375.  DOI: 10.3969/j.issn.1004-5619.2017.04.008 Abstract ( 747 )   Objective To observe and analyze the performance of forensic science in the cases of suxa- methonium chloride poisoning, and to improve the identification of suxamethonium chloride poisoning. Methods Fifty-four cases of suxamethonium chloride poisoning were collected. The rules of determination of suxamethonium chloride poisoning were observed by the retrospective analysis of pathological and toxicological changes as well as case features. Results The pathological features of suxamethonium chloride poisoning were similar to the general changes of sudden death, which mainly included acute pulmonary congestion and edema, and partly showed myocardial disarray and fracture. Suxamethonium chloride could be detected in the heart blood of all cases and in skin tissue of part cases. Conclusion Suxa- methonium chloride poisoning has the characteristics with fast death and covert means, which are difficult to rescue and easily miss inspection. For the cases of sudden death or suspicious death, determination of suxamethonium chloride should be taken as a routine detection index to prevent missing inspection. Related Articles | Metrics

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Application of Susceptibility Weighted Imaging in the Identification of Hemorrhagic Diffuse Axonal Injury LI WEI 2017, 33(4): 376-379.  DOI: 10.3969/j.issn.1004-5619.2017.04.009 Abstract ( 408 )   Objective To explore the application value of susceptibility weighted imaging （SWI） in the forensic identification of hemorrhagic diffuse axonal injury （DAI）. Methods The forensic identification materials of 20 DAI cases were analyzed retrospectively. The sequence image materials of T1WI, T2WI, diffusion weighted imaging （DWI）, fluid attenuated inversion recovery （FLAIR） and susceptibility weighted imaging （SWI） were analyzed statistically. Results DAI related hemorrhage focus commonly located in the superficial area of brain. The detection rate of hemorrhage focus by SWI was the highest, which showing significant difference （P＜0.05） compared with other sequence images. Conclusion The SWI plays an important role in the forensic identification of DAI. Related Articles | Metrics

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DNA Extraction from Old Bones by AutoMate ExpressTM System LI BIN, 吕ZHENG 2017, 33(4): 380-382.  DOI: 10.3969/j.issn.1004-5619.2017.04.010 Abstract ( 549 )   Objective To establish a method for extracting DNA from old bones by AutoMate ExpressTM system. Methods Bones were grinded into powder by freeze-mill. After extraction by AutoMate ExpressTM, DNA were amplified and genotyped by Identifiler?誖Plus and MinFilerTM kits. Results DNA were extracted from 10 old bone samples, which kept in different environments with the postmortem interval from 10 to 20 years, in 3 hours by AutoMate ExpressTM system. Complete STR typing results were obtained from 8 samples. Conclusion AutoMate ExpressTM system can quickly and efficiently extract DNA from old bones, which can be applied in forensic practice. Related Articles | Metrics

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Genetic Polymorphisms of 17 Y-STR Loci in Nantong Han Population YANG MIN, XU SI-XIAN, QIN HAI-YAN, ET AL. 2017, 33(4): 383-387.  DOI: 10.3969/j.issn.1004-5619.2017.04.011 Abstract ( 683 )   Objective To analyze the genetic phenotypes of Nantong Han population and evaluate the application value of 17 Y-STR loci in Nantong population. Methods The peripheral blood samples were collected from 343 unrelated Nantong Han males and the genomic DNA were extracted by Chelex-100 method. Genotyping was performed using the AmpF?詛STR YfilerTM Kit. The results were compared with other 12 Han populations, including Anhui, Jiangsu, Jiangxi, Shandong, Shanghai, Zhejiang （1）, Lanzhou, Nanyang, Luzhou, Mudanjiang, Shanxi and Zhejiang （2）, and 9 minority populations （Mongol, Xibe, Tibetan in Lhasa, Tibetan in Qinghai, Kazak, Uighur, Manchu, Paiwan in Taiwan and Tujia）. Results A total of 327 different haplotypes were found in 17 Y-STR loci in Nantong Han population. The haplotype diversity （HD） was 0.999 7. The Rst value between Nantong Han and other Chinese populations ranged from -0.000 6 to 0.263 5. The multidimensional scaling results showed that Nantong Han population had no significant differences between most of the Han populations, but had significant differences between most of Chinese minority populations. Conclusion Seventeen Y-STR loci can be a powerful tool for forensic application because of its high polymorphism in Nantong Han population. Related Articles | Metrics

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Polymorphisms of 19 STR Loci in Guizhou Han Population and Their Forensic Application ZHAO QIN-SONG, REN ZHENG, ZHANG HONG-LING, DENG 2017, 33(4): 388-392.  DOI: 10.3969/j.issn.1004-5619.2017.04.012 Abstract ( 521 )   Objective To investigate the allelic distribution of 19 autosomal STR loci in Guizhou Han population, and to estimate the forensic application value. Methods The 19 autosomal STR loci in 520 unrelated healthy individuals from Guizhou Han population were studied using GoldeneyeTM 20A kit. The 310 genetic analyzer was used for capillary electrophoresis, and the GeneMapper?誖ID v3.1 for genotyping. Results The heterozygosis, the discrimination power, the probability of exclusion, the polymorphism information content, the cumulative discrimination power and the cumulative probability of exclusion of the 19 STR loci were 0.603 8-0.916 4, 0.790 0-0.985 6, 0.295 5-0.826 9, 0.553 5-0.908 9, 1-1.230 0×10-22 and 0.999 999 99, respectively. Compared with other five Han populations in pairwise allelic frequencies, Guizhou Han only had significant differences with Shandong Han, Liaoning Han and Shanxi Han. Conclusion The 19 autosomal STR loci such as D19S433 have a highly genetic polymorphic in Guizhou Han population, which have application values in the researches of population genetics and forensic genetics. Related Articles | Metrics

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Application of IMA and H-FABP in Forensic Diagnosis of Sudden Cardiac Death ZHU ZE-LEI, WANG PENG, YOU JIA-BIN, ET AL. 2017, 33(4): 393-396.  DOI: 10.3969/j.issn.1004-5619.2017.04.013 Abstract ( 566 )    Acute myocardial ischemia is the most common cause of sudden cardiac death. The diagnosis of early myocardial ischemia is a hot point in forensic medicine, which is also an early and important part for a prevention against myocardial infarction. This paper conducts a comprehensive discussion of the structure, function, clinical value and forensic medicine application prospect of ischemia modified albumin （IMA） and heart-type fatty acid binding protein （H-FABP), aiming to determine whether the two proteins can be used as biochemical detection indicators of early myocardial ischemia for the diagnosis of sudden cardiac death in forensic medicine. Related Articles | Metrics

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Application of Metagenomics in Forensic Identification SA RI-NA, CAI LING-YI, WU HUI-JUAN, ET AL. 2017, 33(4): 397-401.  DOI: 10.3969/j.issn.1004-5619.2017.04.014 Abstract ( 691 )   With the development of molecular biology and genomics, metagenomics is playing a more important role in forensic science and forensic identification. In recent years, as a branch discipline studying the composition profile and diversity of microbe flora as well as studying the interaction within microbe and with environment, the application of metagenomics has gradually risen and brought new opportunities for forensic identification-related area. In this review, strategy of metagenomics and its application in forensic identification including individual identification, origin determination of biological stain in crime scene and drug abuse detection are summarized. This article aims to elucidate the role and application value of metagenomics in forensic science. Related Articles | Metrics