Progress in the Application of Magnetic Nanoparticles in Forensic Trace Analysis / 法医学杂志

Given the complexity of biological samples and the trace nature of target materials in forensic trace analysis, a simple and effective method is needed to obtain sufficient target materials from complex substrates. Magnetic nanoparticles (MNPs) have shown a wide range of application value in many research fields, such as biomedicine, drug delivery and separation, due to their unique superparamagnetic properties, stable physical and chemical properties, biocompatibility, small size, high specific surface area and other characteristics. To apply MNPs in the pretreatment of forensic materials, maximize the extraction rate of the target materials, and minimize interference factors to meet the requirements of trace analysis of the target materials, this paper reviews the application of MNPs in the fields of forensic toxicological analysis, environmental forensic science, trace evidence analysis and criminal investigation in recent years, and provides research ideas for the application of MNPs in forensic trace analysis.

Research Progress on Accumulated Degree Days for PMI Estimation / 法医学杂志

Among many factors that affect postmortem interval （PMI）, temperature is undoubtedly the most important factor, but simply considering temperature in estimation of PMI has limitations. Accumulated degree days （ADD） is an important indicator related to biological growth and development, which considers two major variables, environmental temperature and elapsed time. Therefore, the establishment of mathematical model of ADD and some evaluation indicators to estimate PMI, is obviously better than single temperature variable. This article briefly describes the research history of ADD, its application in PMI estimation, and the problems it faces, to provide reference for the further development of ADD to improve the accuracy of PMI estimation.

Sequential Changes of Total Adenine Nucleotide and Adenylic-Acid Energy Charge in Muscles of Rats after Death / 法医学杂志

Objective To determine the purine adenylate [adenosine triphosphate （ATP）, adenosine diphosphate （ADP）, adenosine monophosphate （AMP）] content in the muscles of both hind limbs of rats at different postmortem interval （PMI）, calculate the changes in the total adenine nucleotide （TAN） content and the adenylic-acid energy charge （AEC）, and explore their relationship with PMI. Methods Healthy rats were sacrificed by cervical dislocation and kept at 20 ℃. The muscles of their hind limbs were extracted at 0, 24, 48, 72, 96, 120, 144, and 168 h after death. Reversed-phase high performance liquid chromatography was used to determine the content of purine adenylates, the TAN and AEC of the muscles of the both hind limbs were calculated, and the related regression equations of their relationship with PMI were established. Results Within 168 h of death of rats, the trend of ATP change was different from ADP, and the content of AMP continuously increased. The TAN value gradually increased with the extension of PMI, and the AEC showed a downward trend within 168 h after death. Among them, the patterns of AEC changes with PMI were obvious, the correlation coefficient was high （ R2=0.903）, and the curve fitting relationship was good; the fitting relationship between ATP, ADP, AMP, TAN and PMI was poor （ R2=0.198-0.754）. Conclusion The postmortem change patterns of AEC provide new research ideas for PMI estimation in the forensic field.

Research Progress on the Determination of Electrical Conductivity in Forensic Science / 法医学杂志

Electrical conductivity （EC） is an important physical and chemical index in electrochemical analysis. In recent years, with the penetration and reference of transformation medicine and interdisciplinary theory and technology in the forensic field, new applications of EC in the field of forensic science have been developed. This paper reviews three aspects of the application of EC, the determination of biological tissue freshness, postmortem interval estimation and the application in forensic taphonomy, in order to provide reference for relevant scientific research and related practices.

Relationship between Electrical Conductivity and Chemical Content of Rat Skeletal Muscle Impregnating Solution and Postmortem Interval / 法医学杂志

Objective To study the mechanism of change of the electrical conductivity （EC） of rat skeletal muscle impregnating solution that occurs with the change of postmortem interval （PMI）. Methods Healthy Sprague-Dawley rats were killed and kept at about 25 ℃. Skeletal muscles were extracted at different PMI--immediate （0 d）, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, and 7 d, then mixed with deionized water to make impregnating solution with a mass concentration of 0.1 g/mL. The solution's EC and nine common chemicals in it, such as potassium ion, calcium ion, and chloride ion, were determined. Results EC increased gradually with the extending of PMI （P=0.024） during the 7 days after the rats' death. The content of uric acid （P=0.032）, urea nitrogen （P=0.013） and phosphorus （P=0.022） also increased during the extension. However, the content of magnesium ions decreased with extending of PMI （P=0.047）. The correlation between potassium ion, sodium ion, chlorine ion, calcium ion, creatinine and PMI were weak （P>0.05）. Conclusion The molecular basis of skeletal muscle EC change in rats after their death is the changes of uric acid, urea nitrogen, inorganic phosphorus and other chemical components. Furthermore, combine use of various indicators can improve the accuracy of the EC method to infer PMI.

Analysis of 28S rRNA and COⅠ Gene Sequence of Nine Necrophagous Calliphorid Flies from Luoyang / 法医学杂志

Objective To assess the feasibility of using 28S ribosomal RNA （28S rRNA） and mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） gene sequences of nine necrophagous Calliphorid flies for the identification of common necrophagous Calliphorid flies, and to provide technical support for postmortem interval （PMI） estimation. Methods Twenty-three Calliphorid flies were collected and identified morphologically, and DNA were extracted from legs. The gene fragments of 28S rRNA and COⅠ were amplified and sequenced, then the sequence alignment was performed with BLAST. The composition of obtained sequences was analyzed and evolutionary divergence rate between species and intraspecies were established. The phylogeny tree was constructed with neighbor-joining method. Results The 23 necrophagous Calliphorid flies were identified to 9 species of 5 genera. The 715 bp from 28S rRNA and 637 bp from COⅠ gene were obtained and the online BLAST result showed more than 99% of similarity. The phylogeny tree showed that the necrophagous flies could cluster well into 9 groups, which was consistent with morphological identification results. The intraspecific difference in 28S rRNA was 0 and the interspecific difference was 0.001-0.033. The intraspecific difference in COⅠ was 0-0.008 and the interspecific difference was 0.006-0.101. Conclusion Combined use of 28S rRNA and COⅠ gene sequence fragments can effectively identify the nine Calliphorid flies in this study. However, for closely related blowfly species, more genetic markers should be explored and used in combination in future.

Relationship between Early Postmortem Interval and Electrical Conductivity of Rat Liver, Spleen and Kidney after Death in Rats / 法医学杂志

OBJECTIVES@#To determine the electrical conductivity （EC） of the liver, spleen and kidney of rats at different postmortem intervals （PMIs） within 24 hours for investigating the relationship between EC of different organs and early PMI.@*METHODS@#Totally 45 SD rats were sacrificed by cervical dislocation and kept at a constant temperature of 25 ℃. Tissues were taken from the liver, spleen, and kidney of rats at 0, 3, 6, 9, 12, 15, 18, 21 and 24 h. Impregnating solution with a mass concentration 0.1 g/mL was prepared using deionized water. The EC value of impregnating solution with different organs was separately determined. The regression equations of EC and PMI for different organs were established, respectively. The relationship between EC of different organs and early PMI was analysed in deceased rats.@*RESULTS@#The relationship between PMI and EC of the liver and spleen was well fitted with the linear equation. The liver showed the best fitting degree followed by the spleen, while the EC of the kidney showed no significant changes within 24 h. There was a good linear relationship between early PMI and the EC of the liver and spleen.@*CONCLUSIONS@#A good linear relationship between early PMI and the EC of the liver and spleen can be found in rats after death, which can be used for the early PMI estimation.

Molecular Identification of Sarcosaphagous Flies in Luoyang Based on 28S rRNA Gene Sequence / 法医学杂志

Objective To detect 715 bp sequence of 28S rRNA in sarcosaphagous flies, and to identify their common species for solving the problem of morphological identification, as well as providing technical support for postmortem interval (PMI) estimation. Methods Twenty-nine common sarcosaphagous flies were collected in Luoyang and classified by morphological characteristics. The DNA was extracted from the fly's legs by Chelex-100 method and then the fragments of 28S rRNA were amplified and sequenced. The results were compared with twenty-eight corresponding fly species of GenBank and EMBL databases. All the sequences were analyzed by MEGA7.0 software, and sequence alignment was performed by the searching in BLAST. The nucleotide composition was analysed, and the intraspecific and interspecific ge-netic distance and phylogenetic tree were established. Results Twenty-nine sarcosaphagous flies were classified into 6 species of 5 genera, 3 families by morphological characteristics. In the obtained 715 bp sequence of 28S rRNA , the comparison result of online BLAST showed that the similarity was 100％. Five species were well clustered by a phylogenetic tree. Between different groups, the interspecific and intraspecific differences ranged from 0.007 to 0.045 and 0 to 0.001, respectively. Conclusion The 28S rRNA target gene sequences shows a good identification capability, which can be a new genetic marker for the identification of sarcosaphagous flies.

Relationship between Electrical Conductivity and Decomposition Rate of Rat Postmortem Skeletal Muscle / 法医学杂志

OBJECTIVES@#To analyze the relationship among electrical conductivity （EC）, total volatile basic nitrogen （TVB-N）, which is an index of decomposition rate for meat production, and postmortem interval （PMI）. To explore the feasibility of EC as an index of cadaveric skeletal muscle decomposition rate and lay the foundation for PMI estimation.@*METHODS@#Healthy Sprague-Dawley rats were sacrificed by cervical vertebrae dislocation and kept at 28 ℃. Muscle of rear limbs was removed at different PMI, homogenized in deionized water and then skeletal extraction liquid of mass concentration 0.1 g/mL was prepared. EC and TVB-N of extraction liquid were separately determined. The correlation between EC (x₁) and TVB-N (x₂) was analyzed, and their regression function was established. The relationship between PMI (y) and these two parameters were studied, and their regression functions were separately established.@*RESULTS@#The change trends of EC and TVB-N of skeletal extraction liquid at different PMI were almost the same, and there was a linear positive correlation between them. The regression equation was x₂=0.14x₁-164.91(R²=0.982). EC and TVB-N of skeletal muscle changed significantly with PMI, and the regression functions were y=19.38x₁³-370.68x₁²+2 526.03 x₁-717.06(R²=0.994), and y=2.56x₂³-48.39x₂²+330.60x₂-255.04(R²=0.997), respectively.@*CONCLUSIONS@#EC and TVB-N of rat postmortem skeletal muscle show similar change trends, which can be used as an index for decomposition rate of cadaveric skeletal muscle and provide a method for further study of late PMI estimation.

Research Progress on Forensic Entomotoxicology / 法医学杂志

Forensic entomotoxicology is a branch of forensic medicine, which applies entomology, toxicology and other related studies to solve the poisoning cases. It has an obvious advantage in the investigation on poisoning death. Based on the expounding definition and research of entomotoxicology, this paper reviews research progress and application value in some aspects of forensic medicine, such as the effects of drugs/toxins on the growth and development of sarcosaphagous insects and the qualitative and quantitative analysis of the drugs/toxins in the poisoned body tissue.

One-step methylation variable position analysis technology in single-tube / 法医学杂志

OBJECTIVE@#To develop the single-tube one-step methylation variable position (MVP) analysis technology-single-tube post-digestion PCR-melting curve analysis (PDP-MCA).@*METHODS@#Based on differentially methylated region (DMR) reported previously as the model, a set of primers with different melting temperatures of products in the two sides of MVP were designed. By using the FastDigest methylation-sensitive restriction enzyme (MSRE), DNA digestion, multiplex amplification, MCA detection and MCA profiles were performed in a single reaction tube. Same samples (peripheral venous blood, semen, and vaginal fluid, 5 samples each type) were tested by single-tube one step MVP and traditional MSRE-PCR MCA technology. To verify the feasibility of this method, the results were compared with that of the traditional technology. The MCA/HRM profiles of different samples were analyzed and compared.@*RESULTS@#When the melting temperature of the fragments had a differential of 2 degrees C, the MCA melting peaks separated well, and MCA detection after multiplex amplification was successful. The single-tube PDP-MCA assay was developed, which integrated multiple reactions (digestion, amplification and detection) into one tube. By this method, the sample-specific profiles and data were analyzed in 2 h, which is similar to that of the traditional method. The rapid classifications of the samples were also realized.@*CONCLUSION@#Multiplex MVPs can be analyzed in a single closed-tube. The single-tube PDP-MCA technology is a simple, fast, and automatable method. It can be used for detection of DNA methylation variations.

Advances in identification of semen stains / 中华男科学杂志

Stain identification has long been a task in forensic biology. The identification of semen stain, one of the most common human stains, can provide crucial information for crime scene reconstruction and forensic investigation. Traditional detection of semen stain depends largely on the microscopic identification of spermatozoa, enzyme activity-based methods or antigen-antibody reactions. These morphological, proteinological and zymological approaches, however, are apparently inadequate in identifying tiny, admixed, degraded or contaminated samples. With the development of transcriptomics and epigenetics, many semen-specific mRNA markers, such as protamine-1 (PRM1) and -2 (PRM2), have been applied to semen and semen stain identification. Messenger RNA profiling shows great promise in identifying tissues as demonstrated by the recognition of specific markers. Further more, studies on tis-sue-specific differential DNA methylation will provide a scrumptious way of identifying difficult samples.

Applications of Alu family in forensic DNA analysis / 法医学杂志

Alu family is the primate specific short interspersed repetitive elements (SINEs). Its abundance and diversity distribution in genome, high methylation level and polymorphic for insertion make them ideally suitable as tools in forensic applications. The application of A4 lu sequence in forensic genomics, include DNA quantitation, race determination, species and gender identification, personal identification, paternity testing and whole-genome amplification. The principles and characteristics of these Alu-based techniques are also summarized. The prospect of Alu as forensic molecular marker is discussed as well.

Retrospective analysis of autopsy on 49 cases of medical tangles in perinatal period / 法医学杂志

Medical tangles caused by the death of women and infants in perinatal period are very normal in the forensic appraisal. The author collected and analyzed 49 cases of these tangles from many aspects, such as sex and age of the dead, hospital,information of autopsy, fault of medical action and so on,and discovered the normal causes of death, medical action's effects and the causes of tangle. It would be useful to the forensic appraisal, settlement and prevention of these medical tangles.