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    Original Articles
    Temporal Expression of NETosis Marker CitH3 in Deep Vein Thrombosis in Mice
    Qian WANG, Song-min YANG, Juan-juan WU, Yu ZHANG, Xiang-meng WANG, Gang CHEN, Peng-fei JIANG
    2025, 41(3): 201-207.  DOI: 10.12116/j.issn.1004-5619.2024.441004
    Abstract ( 86 )   HTML ( 2 )   PDF (2076KB) ( 109 )  

    Objective To detect the expression changes of citrullinated histone H3 (CitH3) during the development of deep vein thrombosis (DVT) in mice, and to explore its value in estimating the time to thrombosis. Methods The inferior vena cava (IVC) of mice was ligated to establish a thrombosis model induced by congestion. Mice were sacrificed under excessive anesthesia at 0 h, 1 d, 3 d, 5 d, 7 d, 10 d, 14 d and 21 d after the modeling, respectively. The congested IVC segments (0 h after modeling) and the thrombosed IVC segments (1-21 days after modeling) were extracted. Immunohistochemistry and double immunofluorescence staining were used to observe the number of neutrophils and the expression of CitH3 during thrombosis. Western blotting was used to detect the protein expression level of CitH3. Results During thrombosis, CitH3 was mainly expressed in neutrophils within the thrombus. A small number of neutrophils and a few CitH3-positive cells were observed at 0 h after modeling in the congested IVC. Between 1 d and 21 d after modeling, the number of neutrophils reached a peak at 1 d and gradually decreased. The number of CitH3-positive cells and their ratio to neutrophils began to increase at 1 d, reached a peak at 5 d after modeling, and then decreased. The expression level of CitH3 protein began to increase at 1 d and reached a peak at 5 d after modeling. Conclusion The expression of CitH3 during DVI shows temporal changes, and is expected to become a biological marker for estimating the formation time of thrombosis.

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    Dual-Channel Shoulder Joint X-ray Bone Age Estimation in Chinese Han Adolescents Based on the Fusion of Segmentation Labels and Original Images
    Hui-ming ZHOU, Dan-yang LI, Lei WAN, Tai-ang LIU, Yuan-zhe LI, Mao-wen WANG, Ya-hui WANG
    2025, 41(3): 208-216.  DOI: 10.12116/j.issn.1004-5619.2025.250106
    Abstract ( 65 )   HTML ( 1 )   PDF (1243KB) ( 98 )  

    Objective To explore a deep learning network model suitable for bone age estimation using shoulder joint X-ray images in Chinese Han adolescents. Methods A retrospective collection of 1 286 shoulder joint X-ray images of Chinese Han adolescents aged 12.0 to <18.0 years (708 males and 578 females) was conducted. Using random sampling, approximately 80% of the samples (1 032 cases) were selected as the training and validation sets for model learning, selection and optimization, and the other 20% samples (254 cases) were used as the test set to evaluate the model’s generalization ability. The original single-channel shoulder joint X-ray images and dual-channel inputs combining original images with segmentation labels (manually annotated shoulder joint regions multiplied pixel-by-pixel with original images, followed by segmentation via the U-Net++ network to retain only key shoulder joint region information) were respectively input into four network models, namely VGG16, ResNet18, ResNet50 and DenseNet121 for bone age estimation. Additionally, manual bone age estimation was conducted on the test set data, and the results were compared with the four network models. The mean absolute error (MAE), root mean square error (RMSE), coefficient of determination (R2), and Pearson correlation coefficient (PCC) were used as main evaluation indicators. Results In the test set, the bone age estimation results of the four models with dual-channel input of shoulder joint X-ray images outperformed those with single-channel input in all four evaluation indicators. Among them, DenseNet121 with dual-channel input achieved best results with MAE of 0.54 years, RMSE of 0.82 years, R2 of 0.76, and PCC (r) of 0.88. Manual estimation yielded an MAE of 0.82 years, ranking second only to dual-channel DenseNet121. Conclusion The DenseNet121 model with dual-channel input combined with original images and segmentation labels is superior to manual evaluation results, and can effectively estimate the bone age of Chinese Han adolescents.

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    Bibliometric and Visual Analysis of Forensic Research on Body Fluid Identification
    Bao-yan XIE, Ruo-cheng XIA, Ting-ting JIANG, Rui-yang TAO, Cheng-tao LI
    2025, 41(3): 217-227.  DOI: 10.12116/j.issn.1004-5619.2024.541105
    Abstract ( 76 )   HTML ( 1 )   PDF (1896KB) ( 117 )  

    Objective To analyze the literature in the field of body fluid identification collected in the Web of Science Core Collection (WoSCC) database from 2000 to 2023, and explore the research status, hotspots and development trends in this field. Methods The CiteSpace software was utilized to conduct a visual analysis of the literature in the field of body fluid identification included in the WoSCC database from 2000 to 2023. Meanwhile, a bibliometric analysis of the annual publication volume, journal distribution, national contribution, research institutions, author collaboration, and keywords of the literature was conducted. Results A total of 715 papers on forensic body fluid identification were included, and the annual publication volume showed a continuous and stable growth. Among the 55 countries (regions) that published papers, the United States ranked first with 174 papers, followed by China with 107 papers. In terms of journal distribution, Forensic Science International: Genetics had the largest number of papers, which accounted for 20% of the total papers. In terms of author collaboration, a total of 2 079 authors participated in body fluid identification research, and the author collaboration network showed a clearly clustered distribution. The keywords analysis revealed that research hotspots focused on traditional methods, specific RNA molecular markers, DNA methylation, spectroscopy, and the application of microbiomics. Conclusion Research in the field of forensic body fluid identification is thriving, and research institutions and teams should strengthen their collaboration. Establishing unified result interpretation standards and systems and exploring the multiple biomarkers combined application methods will be the research hotspots and important directions for future research in this field.

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    Analysis of Genetic Structure among Different Populations Based on 13 Auto-somal STR Loci in CODIS Core
    Xi HE, Zhen TANG, Ming-ying XIA, Yi-qi ZHAO, Yu-ran LUO, Shi-lin LI
    2025, 41(3): 228-236.  DOI: 10.12116/j.issn.1004-5619.2022.521002
    Abstract ( 59 )   HTML ( 3 )   PDF (1120KB) ( 80 )  

    Objective To investigate the genetic differences among different populations based on 13 autosomal STR loci in CODIS core. Methods Data of 13 autosomal STR loci (CSF1PO, FGA, THO1, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11) were collected from 95 populations in scientific journals between 1999 and 2021, soursed from the PubMed database, which had been published. Allele frequencies of loci were sorted out and forensic genetic parameters including gene differentiation coefficient (Gst), total heterozygosity (Ht), subpopulation heterozygosity (Hs) values, and Nei’s DA genetic distance were calculated. Principal component analysis, phylogenetic tree, and multidimensional scale analysis were conducted to assess population genetic structure. Results A total of 265 alleles were detected at the 13 STR loci in these 95 populations. The mean values of Gst, Ht, and Hs were 0.023 247, 0.797 915 and 0.779 365. Population genetic analyses reflected significant differences among populations from Asia, Africa and Europe. In Asian populations, there was a certain degree of distinction between mainland and island populations; the Han population showed a certain degree of distinction with surrounding populations in mainland; while within the Han population, there were two distinct clusters formed by the northern Han and the southern Han. Conclusion The 13 autosomal STR loci in CODIS core demonstrate potential value for population identification across different groups, and may be used for the differentiation of ethnic groups, among different continental populations.

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    Dental Floss-derived Biological Sample Collection, DNA Extraction and STR Typing
    Ze-qin LI, Fang YUAN, Na LIU, Jiang-wei YAN, Geng-qian ZHANG
    2025, 41(3): 237-243.  DOI: 10.12116/j.issn.1004-5619.2024.540404
    Abstract ( 113 )   HTML ( 4 )   PDF (783KB) ( 226 )  

    Objective To evaluate the forensic application value of used dental floss as a source of biological evidence for individual identification by analyzing the effects of dental floss sample collection methods, DNA extraction methods, preservation conditions, and sampling sites on the success rate of STR typing. Methods Dental floss samples were collected using three techniques: direct cutting, cotton swab wiping, and flocked swab wiping, respectively. DNA was extracted respectively by the Chelex, spin column-based and magnetic bead-based methods. DNA quantification and STR typing were performed using the Qubit kit and FGI HumDNA Typing kit (Platinum), respectively. Storage environments (temperature and humidity, ultraviolet radiation) and sampling locations (the floss part, the handle part) on DNA quantity and STR typing were evaluated. Results Through conducting a statistical analysis of three key indicators of average DNA mass concentration, STR locus detection rate, and typing accuracy rate, the direct cutting method demonstrated the highest efficacy, followed by cotton swab wiping mothed, and the flocked swab wiping method had the lowest efficacy. Direct cutting yielded an average DNA mass concentration greater than (4.94±1.87) ng/μL, with STR locus detection and accuracy rates of 100%. Bead-based DNA extraction method produced superior DNA concentration and quality compared to spin column-based and Chelex methods, regardless of whether the sampling technique used. Preservation conditions had a significant impact on the DNA analysis of samples. Particularly, the STR typing accuracy of samples preserved at 55 ℃/50%RH for 35 days dropped to (81.82±12.31)%, and that of samples exposed to ultraviolet radiation for 12 h dropped to (55.46±34.31)%. DNA concentration from the handle part of dental floss was extremely low, with an STR typing accuracy of only (30.91±27.35)%. Conclusion Using cotton swabs to wipe or directly cutting the thread of dental floss samples, and combining this approach with the magnetic bead method for DNA extraction, can best guarantee the concentration and quality of DNA. In addition, samples should be stored in low-temperature, low-humidity environment, protected from light and ultraviolet radiation.

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    Techniques and Applications
    Comparison of Three Drowning‑related Plankton Testing Methods in Drowning Diagnosis
    Xiao-feng ZHANG, Qin SU, Xiao-hui CHEN, Wei-bin WU, Dong-yun ZHENG, Jian ZHAO, Ling CHEN, Qu-yi XU, Chao LIU
    2025, 41(3): 244-251.  DOI: 10.12116/j.issn.1004-5619.2024.440606
    Abstract ( 92 )   HTML ( 4 )   PDF (953KB) ( 98 )  

    Objective To compare the application effects of plankton multiplex polymerase chain reaction-capillary electrophoresis (PCR-CE), SYBR Green Ⅰ real-time quantitative PCR (qPCR) and microwave digestion-vacuum filtration-automated scanning electron microscopy (MD-VF-Auto SEM) in the diagnosis of drowning. Methods Lung, liver and kidney tissues from 212 drowned corpses and 30 non-drowned corpses were examined respectively by the three drowning-related plankton testing methods, and the detection rates of plankton in each tissue by three methods were compared. Results In drowned corpses, the total detection rates of PCR-CE, qPCR, and MD-VF-Auto SEM were 93.9%, 96.2%, and 95.3%, respectively, with no statistically significant difference (P>0.05). The detection rate of lung tissue by MD-VF-Auto SEM (100%) was higher than those of PCR-CE and qPCR (P<0.05), and there was no significant difference in the detection rates of the three methods in liver or kidney tissues (P>0.05). In non-drowning corpses, a small number of diatoms (less than 10 cells/10 g) were detected by MD-VF-Auto SEM method, only in liver and kidney tissues, while the other two methods yielded negative results for all tissues. Conclusion All three methods have good efficacy in the examination of drowned corpses. The MD-VF-Auto SEM method directly observes diatom morphological characteristics through scanning electron microscopy, and the qualitative and quantitative analyses are intuitive and accurate. It has great advantages in the examination of difficult degradation samples. The PCR-CE method and qPCR method have a low sample demand (0.5 g), are easy to operate and have short detection time (4-7 h). They are easy to be applied in the grassroots departments and are suitable for the rapid determination of drowned corpses in routin cases. The combination of the two DNA methods with the MD-VF-Auto SEM method can increase the detection rate of plankton, ensuring the reliability of examination results. This combined use is of significant importance in the application of drowning diagnosis.

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    Validation and Forensic Application of a Domestic Human DNA Quantitative Detection Kit
    Jing CHEN, Ya-ping WANG, Yun-peng FENG, Xiao-xin HU, Zhen-jun JIA, Hong-di LIU, An-xin YAN, Yong-jiu LI, Zhu PENG, Zhi-fang LIU, Jian-gang CHEN
    2025, 41(3): 252-259.  DOI: 10.12116/j.issn.1004-5619.2023.531101
    Abstract ( 165 )   HTML ( 3 )   PDF (917KB) ( 252 )  

    Objective To verify the efficacy of a domestic human DNA quantification kit based on real-time fluorescence quantitative PCR in detecting the total human DNA concentration, male DNA concentration in mixed male/female DNA samples, the degree of DNA degradation and inhibitor tolerance. Methods Samples with different concentrations, different male/female ratios, different concentrations of inhibitors, and different degradation degrees were tested using the domestic human DNA quantification kit based on real-time fluorescence quantitative PCR. This kit was compared with a similar product on the market and was applied to the detection of DNA from real cases. Results This human DNA quantification kit can effectively detect human DNA as low as 0.001 65 ng/μL, and 6.25 pg/μL of male DNA in mixed samples with a male-to-female ratio of 1∶15 000. Even when the sample contains as high as 400 ng/μL of humic acid or 1 000 μmol/L of hemin alone, the DNA concentration can still be accurately detected. The degradation index can effectively characterize the degradation degree of the sample. This kit has been successfully applied in forensic practice. Conclusion This human DNA quantification kit is accurate and reliable in detection. It can accurately reflect the degradation of DNA and inhibitor tolerance. It has good performance in quantitative accuracy, determination of the male/female ratio in mixed samples, and inhibitor tolerance. It has application potential in forensic case examination.

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    Reviews
    Application of Forensic Transcriptomics in the Identification of Tissue Origin of Body Fluid Stains
    Yi-fan BAI, He-miao ZHAO, Jing CHEN, Hong-di LIU, Rui-qin YANG, Chong WANG
    2025, 41(3): 260-266.  DOI: 10.12116/j.issn.1004-5619.2024.540306
    Abstract ( 65 )   HTML ( 1 )   PDF (680KB) ( 107 )  

    The inference of tissue origin of body fluid stains is crucial for case investigation and court proceedings. However, traditional methods for identification of body fluid stains, such as morphological, chemical, and immunoassay identifications have certain limitations, and there is an urgent need for more efficient methods for confirmatory experiments. In recent years, the rapid development of transcriptomics technology has provided new means for the identification of tissue origin of body fluid stains. Different types of RNA in the transcriptome have their own advantages. This paper elaborates in detail on the application of different types of RNA, such as mRNA, miRNA, circRNA, lncRNA, piRNA and microbial transcriptomics in body fluid identification, and summarizes their respective advantages and limitations, in order to provide a reference for related research.

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    Development of Benchtop Low‑Field Nuclear Magnetic Resonance Technology and Its Application in Drug Control Field
    Qi LIAO, Yong-hong LIU, Ying JIAO, Xiao-ying YANG, Yi-hua YANG, Cui-mei LIU, Rui-xia GAO
    2025, 41(3): 267-276.  DOI: 10.12116/j.issn.1004-5619.2024.340402
    Abstract ( 42 )   HTML ( 1 )   PDF (947KB) ( 56 )  

    At present, the drug substitutes represented by new psychoactive substances are gradually becoming popular, leading to an increasing demand for identifying novel drugs with unknown structures in drug investigation. Nuclear magnetic resonance (NMR) spectroscopy is an important tool for analyzing molecular structures. In the absence of standard substances, quantitative NMR (qNMR) can undertake the quantitative analysis of target substances in complex mixtures and has unique advantages in the research of new drugs and their precursor drugs. Due to the limitations of the site and maintenance costs, as well as relatively complex operation, high-field superconducting NMR is less commonly applied in drug research. The desktop low‑field NMR developed in recent years provides a new alternative solution. Due to the use of permanent magnets, its size is reduced, and the operation and maintenance costs are lowered. It has been widely used in various research fields. This article reviews the development of low-field NMR technology, summarizes the application of desktop low-field NMR in screening and identification of suspicious substances, rapid content determination, analysis of drug manufacturing processes and synthetic routes, and correlation traceability. It also looks forward to the prospects and development directions of this technology in drug research, aiming to provide a reference for researchers who work in analytical chemistry and drug research.

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