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25 April 2026, Volume 42 Issue 2 Previous Issue   
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Original Articles

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Construction of a Segmented PMI Estimation Model Integrating Intestinal Microbial Signatures and Machine Learning in Nude Mice Xiangyan ZHANG, Fan YANG, Sheng HU, Qiong JIA, Hao NIE, Xingchun ZHAO, Yadong GUO 2026, 42(2): 87-93.  DOI: 10.12116/j.issn.1004-5619.2025.450505 Abstract ( 111 )   HTML ( 5 )   PDF (1558KB) ( 47 )   Objective To observe stage-specific changes in the intestinal microbiota of nude mice after death and to develop a postmortem interval （PMI） estimation model based on “rupture points”, thereby exploring a new model for PMI estimation. Methods A total of 108 nude mice were sacrificed, and cecal contents were collected at 18 time points （0, 24, 41, 48, 55, 65, 72, 79, 89, 96, 103, 113, 120, 144, 168, 192, 216, and 240 h postmortem）. 16S rRNA gene amplicon sequencing was used to analyze the changes in intestinal microbiota. Based on microbial abundance, a random forest model was employed for cross-validation to identify signature bacterial genera. A segmented regression model was then constructed to estimate PMI and compared with a direct regression model. Results Both α- diversity and β-diversity analyses indicated significant changes in the relative abundance of intestinal microbiota during the periods of 0-103 h and 113-240 h postmortem in nude mice. The segmented regression model built using the random forest algorithm achieved an R2 of 0.96 and a mean absolute error （MAE） of 9.83 h for PMI estimation. In contrast, the direct regression model yielded an R2 of 0.81 and an MAE of 16.91 h. Conclusion Microbial succession during cadaver decomposition exhibits clear temporal and stage-specific characteristics. A segmented regression model for PMI estimation using “rupture points” can improve the accuracy of PMI estimation in nude mice. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Bioinformatics-Based Screening and Validation of Biomarkers for Sudden Death in Leukemia Bowen LIU, Zhong ZHANG, Feng ZHANG, Yanyan FAN, Peilong LAI 2026, 42(2): 94-101.  DOI: 10.12116/j.issn.1004-5619.2026.460406 Abstract ( 83 )   HTML ( 4 )   PDF (1810KB) ( 38 )   Objective To identify biomarkers for sudden death in leukemia through common genes across leukemia subtypes using bioinformatics technology, and to validate them in a specific case repository of sudden death in leukemia, thereby providing reference for cause of death determination in leukemia-related sudden death and cause of death analysis in medical malpractice cases. Methods Differentially expressed genes （DEGs） common across different types of leukemia were identified using the GEO database （GSE13159 dataset）. Enrichment analyses were performed using the Gene Ontology （GO） and the Kyoto Encyclopedia of Genes and Genomes （KEGG） databases, and enriched pathways were mapped to routine clinical laboratory parameters. Case data from patients who died suddenly due to stroke, respiratory failure, sudden cardiac death, and infectious shock, were collected and divided into the case group（leukemia sudden death group） and the control group（non-leukemia sudden death group） for validation. The Wilcoxon rank sum test and univariate logistic regression were used to screen biomarkers. A multivariate logistic regression model was constructed based on univariate logistic regression results to predict whether a sudden death patient had leukemia. Model performance was evaluated using receiver operator characteristic （ROC） curves and area under the curve （AUC）. Results A total of 540 DEGs shared across four leukemia subtypes were significantly enriched in pathways related to cell cycle, inflammatory immunity, and coagulation. The pathway enrichment results showed clear correspondence with routine clinical laboratory parameters. Compared with the control group, the case group exhibited overall characteristics of low white blood cell counts （mean: 2.42×109/L）, low platelet counts （mean: 23×109/L）, and low interleukin-6 （IL-6） levels （mean: 386.0 pg/mL）. Based on the univariate logistic regression results, IL-6 and platelet count were included in a multivariate logistic regression model. The model yielded an AUC of 0.828. Conclusion The enrichment pattern of DEGs in leukemia sudden death biomarkers is consistent with the shared molecular mechanisms of uncontrolled leukemia cell proliferation, immune evasion, and hypercoagulable state. The combination of blood routine test parameters and IL-6 can be used for the differential diagnosis of sudden death in leukemia, providing objective evidence for auxiliary determination of cause of death in leukemia sudden death cases and offering a quantitative reference for cause of death analysis in medical malpractice identification. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Construction of a Multimodal Forensic Pathology Database and Implementation of Model-Augmented Retrieval Haojie QIN, Enhao FU, Yongchao YANG, Yawen LIU, Mingzhu JIA, Zhiling TIAN, Zhe ZHENG, Siyun GUO, Ningguo LIU 2026, 42(2): 102-111.  DOI: 10.12116/j.issn.1004-5619.2025.451106 Abstract ( 101 )   HTML ( 2 )   PDF (2113KB) ( 83 )   Objective To construct a multimodal forensic pathology database based on artificial intelligence （AI） technology and explore methods for integrating pre-trained models into multimodal databases. Methods A hybrid storage architecture consisting of MySQL, Redis and OSS was employed to manage desensitized multimodal data. Data entry was optimized using optical character recognition （OCR） and natural language processing （NLP） technologies. OCR performance was evaluated and optimized using cosine distance, character error rate （CER）, and word error rate （WER） to meet practical operational requirements. An intelligent retrieval model was developed using the ChatGLM3-6B model and retrieval-augmented generation （RAG） technology. Model performance was evaluated using ranking metrics including Precision@K, Recall@K, discounted cumulative gain （DCG）, and normalized discounted cumulative gain （NDCG）. Results The database demonstrated satisfactory baseline performance. The response times were maintained within 150 ms for single-query condition and within 2 s for multiple-query conditions. The average disk read throughput reached 950 MB/s. In concurrent performance tests, the database achieved a maximum throughput of 1 200 queries per second （QPS）, meeting multimodal data management demands. OCR evaluation showed high recognition accuracy; for high-quality documents, the cosine distance, CER, and WER achieved 0.02, 1.5%, and 3.2%, respectively. Intelligent retrieval results indicated that Precision@K remained consistently high （0.69-1.00）, while NDCG values remained above 0.87 for all evaluations. When K=100, the NDCG surpassed 0.95 for all queries, meeting expected performance requirements. Conclusion The multimodal forensic pathology database constructed in this study demonstrates good stability and operational efficiency and can meet the requirements of routine forensic practice for multimodal data storage, management, and analysis. The intelligent retrieval capabilities, based on pre-trained large language models （LLMs）, can be applied to conversational information retrieval from forensic reports and related documents, providing a novel approach to the management and analysis of multimodal databases. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Effect of RNA Sequencing Normalization on Forensic Age Estimation Jinyuan ZHAO, Yujia XUAN, Anqi CHEN, Yanfang LU, Mengxiao LIAO, Sitong LIU, Yu XING, Yali WANG, Liqin CHEN, Chengtao LI 2026, 42(2): 112-120.  DOI: 10.12116/j.issn.1004-5619.2025.550701 Abstract ( 70 )   HTML ( 4 )   PDF (1188KB) ( 23 )   Objective To evaluate the applicability of four commonly used RNA sequencing normalization methods in forensic age estimation, and to provide a reference for optimizing the accuracy and precision of age estimation models. Methods Peripheral blood samples were collected from 147 unrelated Chinese Han individuals. After RNA sequencing, gene expression levels were normalized using four methods: counts per million （CPM）, fragments per kilobase of transcript per million mapped reads （FPKM）, transcripts per million （TPM）, and trimmed mean of M-values （TMM）. Spearman correlation analysis was used to screen age-related mRNAs. Three age estimation models were constructed using least absolute shrinkage and selection operator （LASSO）, support vector machine （SVM）, and extreme gradient boosting （XGBoost）, respectively, to compare the performance of the four normalization methods. Results The four normalization methods screened different numbers of potential age-related markers. The TMM method screened the largest number of 2 912 markers. The number of potential markers screened was 2 897 and 1 481 for CPM and FPKM, respectively. The TPM method screened the fewest markers with the number of 338. Age estimation models constructed based on these markers exhibited mean absolute errors （MAE） ranging from 4.38 to 8.62 years in the training set and from 5.85 to 9.29 years in the test set. The TMM method performed the best among the three models, especially in the XGBoost model; it achieved an MAE of 4.38 years in the training set and 5.85 years in the test set for the XGBoost model. Conclusion Different normalization methods have a large impact on the construction of forensic age estimation models. The TMM normalization method can effectively improve prediction accuracy and is recommended as the preferred method for forensic age estimation. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Analysis of Abnormal Genotyping at Amelogenin Locus in Male Individuals Zhenping LIU, Zhihua YE, Jijun TONG, Jiahui SONG, Weiwei WU, Honglei HAO, Yanfang FU, Xiandun ZHAI 2026, 42(2): 121-129.  DOI: 10.12116/j.issn.1004-5619.2024.540305 Abstract ( 105 )   HTML ( 4 )   PDF (1736KB) ( 76 )   Objective To investigate the abnormal genotyping and its causes at the Amelogenin locus in male samples. Methods A total of 23 647 blood samples from unrelated male individuals were analyzed using the STRtyper-21G kit, and 38 samples with abnormal Amelogenin locus were identified. These samples were retested and classified using GlobalFilerTM and PowerPlex® 21 kits. Additional sex chromosome STR genotyping and Sanger sequencing were performed for samples with abnormal genotypes. Sequence-tagged site （STS） testing was conducted for samples suspected of Amel-Y microdeletions. Results Among above 38 samples, except for Amelogenin locus, all samples showed normal male sex chromosome STR typing. The detection rate of abnormal genotyping was 0.161% （38/23 647）, which were categorized into three major types. Among them, 30 cases had Amel-X deletion: 5 cases had C→T mutation at position 372; 2 cases had G→A mutation at position 293; 23 cases had A→G mutation at position 304. There were 2 cases of Amel-Y deletion: 1 case of insertion mutation of TTAA at position 387, and 1 case of microdeletion of the short arm containing Amel-Y. Six cases of abnormal Amel-X/Y peak ratios were identified: using the STRtyper-21G kit, the abnormalities appeared as low Amel-X with absent Amel-Y, normal Amel-X with absent Amel-Y, normal Amel-X with low Amel-Y, respectively. However, retesting with GlobalFilerTM and PowerPlex® 21 kits consistently showed normal Amel-X with low Amel-Y. The GlobalFilerTM profiling showed that the peak heights of Amel-Y were comparable to those of the Y-InDel and DYS391 markers, and no abnormalities were detected by sequencing. Conclusion Amelogenin genotyping abnormalities occur at a measurable frequency in the population and are mainly associated with mutations, which can be categorized as Amel-X deletion, Amel-Y deletion, and Amel-X/Y peak ratio abnormality. Regarding normal Amel-X peaks with lower Amel-Y peaks, the possibility of mosaic loss of chromosome Y （mLOY） in samples, which is commonly observed in elderly males, should be considered and given attention. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Detection of Chlormezanone in Blood Using HPLC-MS/MS Method Yue LIU, Weiwei LIANG, Huanhui ZHU, Tianfu HE, Yuanyuan TIAN, Cong PENG, Songcai WANG 2026, 42(2): 130-134.  DOI: 10.12116/j.issn.1004-5619.2024.340403 Abstract ( 95 )   HTML ( 3 )   PDF (826KB) ( 79 )   Objective To establish a high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method for detecting chlormezanone in blood samples. Methods Acetonitrile was added to the samples to precipitate proteins. After vortex and ultrasonication, the mixture was centrifuged, and the supernatant was collected. It was then filtered using a 0.22 μm polytetrafluoroethylene membrane. Separation was performed using an InfinityLab Poroshell 120 EC-C18 column （100 mm×4.6 mm, 2.7 μm）. The mobile phase consisted of phase A （0.1% formic acid） and phase B （acetonitrile）, with gradient elution at a flow rate of 0.4 mL/min. The mass spectrometer was operated with an electrospray ionization in positive ion mode and multiple reaction monitoring mode. Results Chlormezanone in blood samples showed good linearity within the tested range, with the correlation coefficients （r） all greater than 0.999. The limit of detection and the limit of quantitation of chlormezanone were 22.83 ng/mL and 100 ng/mL, respectively. The matrix effects were 2.1%-3.9% and recoveries were 88.8%-92.3%. Using this method, the mass concentration mass of chlormezanone detected in a positive sample was 1 217.23 ng/mL. Conclusion This method requires simple sample preparation and a small sample volume, and offers a wide linear range, making it suitable for the detection of chlormezanone in blood. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

Cases Study

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Forensic Analysis of 173 Cases of Rotator Cuff Injury Caused by Road Traffic Accidents Yaobin WANG, Lili WANG, Daoyin GONG 2026, 42(2): 135-142.  DOI: 10.12116/j.issn.1004-5619.2024.240412 Abstract ( 197 )   HTML ( 16 )   PDF (691KB) ( 97 )   Objective To investigate the epidemiological and forensic characteristics of rotator cuff injuries caused by road traffic accidents, and to explore the timing and influencing factors of disability assessment of rotator cuff injuries. Methods A retrospective analysis was conducted on 173 cases of rotator cuff injury caused by road traffic accidents. Factors analyzed included the time of post-injury assessment, the degree of functional loss of shoulder joint movement in six directions, the treatment methods, the presence or absence of combined shoulder injuries and the type of combined shoulder injuries, and the type of rotator cuff injury. The Kruskal-Wallis test and Wilcoxon rank sum test were used to analyze the differences in functional loss among the six directions of shoulder joint movement. The t-test was used to compare the differences in functional loss between the conservative treatment and the surgical treatment, as well as between cases with and without combined shoulder injuries. One-way analysis of variance （ANOVA） and Student-Newman-Keuls （SNK） test were used to analyze the differences in functional loss of shoulder joint movement among the injury types of contusion, mild tear, moderate tear and severe tear. One-way ANOVA and least significant difference （LSD） test were used to analyze the differences according to the number of involved rotator cuff tendons （1, 2, 3, or 4 tendons） and post-injury assessment time （3-<6 months, 6-<9 months, 9-<12 months, and ≥12 months）. Results The median assessment time after injury was 290.00 days. Among the six directions of shoulder joint movement, statistically significant differences in the degree of functional loss were observed between abduction up, extension back and external rotation compared with adduction, as well as between external rotation and forward flexion up （P<0.05）. There was no statistically significant difference in the degree of functional loss between conservative treatment and surgical treatment, or among different post-injury assessment times （P>0.05）. However, there were significant differences between the non-combined shoulder injury and combined shoulder injury, different types of rotator cuff injuries, and the different numbers of involved rotator cuff tendons （P<0.05）. Treatment methods had no significant effect on the degree of functional loss of shoulder joint movement, whereas combined shoulder injuries, more severe rotator cuff injuries, and involvement of a greater number of rotator cuff tendons were associated with more severe functional loss of shoulder joint movement. Conclusion Rotator cuff injuries caused by road traffic accidents exhibit the epidemiological and forensic characteristics including delayed diagnosis after injury, prolonged post-injury assessment time, and the rotator cuff injuries being mainly mild to moderate tears （≤3 cm） and involving 1 or 2 tendons. Combined shoulder injuries are mostly characterized by greater tuberosity fractures of the humerus, long head of the biceps tendon injuries and shoulder dislocations. The recommended timing for disability assessment in cases involving rotator cuff tears and/or surgical treatment should be appropriately extended. The presence or absence of combined shoulder injuries, the types of rotator cuff injury, and the number of involved rotator cuff tendons should be emphasized in forensic clinical identification. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

Reviews

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Methods， Applications and Prospects of RNA Polymorphism for Individual Identification Gengqian ZHANG, Zidong LIU 2026, 42(2): 143-150.  DOI: 10.12116/j.issn.1004-5619.2024.541104 Abstract ( 132 )   HTML ( 3 )   PDF (673KB) ( 106 )   In recent years, RNA markers have been widely employed for tissue origin identification in forensics. RNA molecules are diverse and can be detected in dried biological stains. They show characteristics such as tissue specificity, spatiotemporal dynamic expression, multi-copy numbers and high polymorphic genetics. Based on these characteristics, RNA genetic markers that combine polymorphism with body fluid specificity have been successfully applied to identify target body fluids in the multi-donor mixed stains. In addition, RNA genetic markers with both multi-copy characteristics and polymorphism have shown promising results in individual identification using anucleate samples such as hair shafts. However, the application of RNA polymorphic genetic markers still faces several key challenges, including the selection of appropriate RNA and polymorphic loci, and optimization of their practical use in forensics. This paper reviews the application of RNA polymorphisms in individual identification and systematically analyzes its advantages, limitations, and the main challenges that need to be addressed in the future. The goals are to further review and consolidate findings from studies of RNA polymorphisms, to advance theoretical development and methodological innovation of RNA polymorphisms in individual identification and to provide references for forensic individual identification. References | Supplementary Material | Related Articles | Metrics

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Research Progress on Mescaline and Its Analogues Ruidi SHEN, Zhenhua QIAN 2026, 42(2): 151-158.  DOI: 10.12116/j.issn.1004-5619.2024.340703 Abstract ( 115 )   HTML ( 4 )   PDF (881KB) ( 132 )   The abuse of new psychoactive substances is severe globally. Among phenethylamine compounds, mescaline and its analogues are a class of substances that are often overlooked, yet their abuse has shown a gradual increase in recent years. This paper reviews mescaline and its analogues from three aspects: their basic information, pharmacological and toxicological properties, and detection methods. The aim is to provide insights for future research on mescaline and its analogues, as well as to provide references for forensic examination involving these substances. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

Education and Administration

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Construction and Practice of the “Leading Goose Navigation and Four-Dimensional Synergy” Model for Cultivating Top Innovative Talents in Forensic Medicine Qingxia LIU, Di WEN, Shujin LI, Chunling MA, Bin CONG 2026, 42(2): 159-163.  DOI: 10.12116/j.issn.1004-5619.2025.451203 Abstract ( 85 )   HTML ( 2 )   PDF (653KB) ( 70 )   To address problems such as fragmented research training, a disconnect between theory and practice, weak value guidance, and a limited international perspective in the traditional training of high-level forensic medicine professionals, the forensic medicine program at Hebei Medical University constructed a new graduate training model titled “Leading Goose Navigation and Four-Dimensional Synergy”. This model leverages the “Leading Goose” effect of high-level academic leaders, such as academicians, to establish strategic focus and a spiritual highland. It reshapes the educational ecosystem by taking research-based education to integrate the innovation chain, practical training to forge professional competence, ideological education to shape professional conviction, and international education to broaden international horizons as specific paths for “Four-Dimensional Synergy”. Using case study and comparative analysis methods, postgraduate cohorts from 2020 to 2023 （N=109） were selected as the post-reform samples and retrospectively compared with data from cohorts between 2015 and 2019. Empirical results showed a 98.6% increase in per capita SCI paper publications and a 16.58% increase in the professional match rate of employment, along with significantly enhanced professional commitment and international perspective. Furthermore, this model demonstrates potential for extension to undergraduate education and evolution through “Forensic Medicine + AI” interdisciplinary integration, providing a valuable reference for cultivating top innovative talents in applied disciplines in the new era. References | Supplementary Material | Related Articles | Metrics

Frontier Express

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Insecticide： Chlorfenapyr Hang CHEN, Xin WANG, Ping XIANG, Hui YAN 2026, 42(2): 164-167.  DOI: 10.12116/j.issn.1004-5619.2026.360304 Abstract ( 78 )   HTML ( 12 )   PDF (607KB) ( 27 )   Chlorfenapyr is a broad-spectrum insecticide and acaricide derived from natural pyrrole derivatives. After entering the body, it is rapidly converted into the active metabolite tralopyril. At present, there is no specific antidote for chlorfenapyr poisoning. The mortality rate associated with this substance is relatively high, because of the latent period following exposure. In recent years, cases of poisoning caused by chlorfenapyr have been increasing. Based on relevant domestic and international research, this paper systematically reviews the toxicological effects, toxicity data, poisoning symptoms, in vivo processes of chlorfenapyr, as well as the toxicological data from typical cases, with the aim of providing a reference for forensic identification. Figures and Tables | References | Supplementary Material | Related Articles | Metrics

Case Reports

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Homicidal Strangulation by Jujitsu “Blood Choke” Disguised as Sudden Death： A Case Report Zhiheng LI, Fan CHEN, Yang LI 2026, 42(2): 168-170.  DOI: 10.12116/j.issn.1004-5619.2024.440808 Abstract ( 296 )   HTML ( 12 )   PDF (1015KB) ( 220 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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column:Indoor Cadaver Damaged by Domestic Dog and Mistakenly Identified as Human Assault： A Case Report Ming PEI, Fei CHEN, Yonghong KANG, Hua XIE, Junhua YANG, Zhengdong LI 2026, 42(2): 171-173.  DOI: 10.12116/j.issn.1004-5619.2024.440505 Abstract ( 372 )   HTML ( 10 )   PDF (860KB) ( 332 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Cardiac Tamponade Death Due to Rupture of Coronary Artery Aneurysm Caused by Incomplete Kawasaki Disease： A Case Report Xinxin GAO, Hanchao YANG, Jing MA, Jiayu CUI, Kaixuan SUN, Baogang ZHANG 2026, 42(2): 173-176.  DOI: 10.12116/j.issn.1004-5619.2025.450306 Abstract ( 162 )   HTML ( 5 )   PDF (1415KB) ( 252 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Homicide Followed by Suicide Due to the Abuse of New Psychoactive Substance 3-MeO-PCPy： A Case Report Xi ZHAO, Xinli SHI, Guipeng WANG, Xin WANG, Junbo ZHAO, Min CHEN, Jianhua ZHANG 2026, 42(2): 177-179.  DOI: 10.12116/j.issn.1004-5619.2025.451002 Abstract ( 232 )   HTML ( 7 )   PDF (541KB) ( 210 )   References | Supplementary Material | Related Articles | Metrics

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Forensic Clinical Identification of Aortic Dissection Complicated with Delayed Intestinal Rupture： A Case Report Fangyu ZHAO, Dong GAO 2026, 42(2): 180-183.  DOI: 10.12116/j.issn.1004-5619.2024.240803 Abstract ( 101 )   HTML ( 8 )   PDF (824KB) ( 115 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Disability Grade Assessment of Non-Limb-Paralysis Movement Disorder Due to Ataxia Caused by Heat Stroke： A Case Report Xia LAN, Yingqi WANG, Saiying XU, Long CHEN, Yiwen SHEN, Jieqing JIANG 2026, 42(2): 184-188.  DOI: 10.12116/j.issn.1004-5619.2025.250901 Abstract ( 63 )   HTML ( 3 )   PDF (872KB) ( 27 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics

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Forensic Identification of Unilateral Separated Hyoid Bone Misdiagnosed as Hyoid Bone Fracture： A Case Report Shiyi WANG, Yusen WANG, Hao XIAO, Jing WANG, Li LAN, Qi WANG 2026, 42(2): 189-190.  DOI: 10.12116/j.issn.1004-5619.2025.250107 Abstract ( 65 )   HTML ( 4 )   PDF (472KB) ( 35 )   Figures and Tables | References | Supplementary Material | Related Articles | Metrics