Research Progress of Exosomes and Their Forensic Significance

doi:10.12116/j.issn.1004-5619.2021.410302

Abstract

Abstract:

Exosomes are membranous tiny vesicles secreted by cells, which are widely found in the extracellular matrix and various body fluids and carry a variety of biologically functional molecules such as proteins, lipids, messenger RNA （mRNA） and microRNA （miRNA）. Exosomes not only play important biological roles in the field of immunology and oncology, but also have potential application value in the field of forensic medicine. This article reviews the discovery, production and degeneration mechanism, biological functions, isolation and identification methods of exosomes, summarizes the research on exosomes and their significance in the field of forensic science, and discusses their applications in body fluid identification, individual identification, postmortem interval estimation to provide ideas for the application of exosomes in forensic work.

Key words: forensic medicine, exosomes, individual identification, estimation of postmortem interval, review

CLC Number:

DF795.1

Yuan-bo KANG, Si-fan WANG, Ting-jun CHEN, Ya-dong GUO, Chang-quan ZHANG. Research Progress of Exosomes and Their Forensic Significance[J]. Journal of Forensic Medicine, 2022, 38(6): 754-762.

Figures/Tables 2

References 77

1	PEGTEL D M， GOULD S J. Exosomes[J]. Annu Rev Biochem，2019，88：487-514. doi：10.1146/annurev- biochem-013118-111902 .
2	ANEL A， GALLEGO-LLEYDA A， DE MIGUEL D， et al. Role of exosomes in the regulation of T-cell mediated immune responses and in autoimmune disease[J]. Cells，2019，8（2）：154. doi：10.3390/cells 8020154 .
3	KALLURI R， LEBLEU V S. The biology， function， and biomedical applications of exosomes[J]. Science，2020，367（6478）：eaau6977. doi：10.1126/sci ence.aau6977 .
4	WOLF P. The nature and significance of platelet products in human plasma[J]. Br J Haematol，1967，13（3）：269-288. doi：10.1111/j.1365-2141.1967.tb087
41	x.
5	HARDING C， HEUSER J， STAHL P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes[J]. J Cell Biol，1983，97（2）：329-339. doi：10.1083/jcb.97.2. 329 .
6	PAN B T， JOHNSTONE R M. Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro： Selective externalization of the receptor[J]. Cell，1983，33（3）：967-978. doi：10.1016/0092-8674（83）90040-5 .
7	JOHNSTONE R M， ADAM M， HAMMOND J R， et al. Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles （exosomes）[J]. J Biol Chem，1987，262（19）：9412-9420.
8	RAPOSO G， NIJMAN H W， STOORVOGEL W， et al. B lymphocytes secrete antigen-presenting vesi-cles[J]. J Exp Med，1996，183（3）：1161-1172. doi：10 .
	1084/jem.183.3.1161.
9	CORRADO C， RAIMONDO S， CHIESI A， et al. Exosomes as intercellular signaling organelles involved in health and disease： Basic science and clinical applications[J]. Int J Mol Sci，2013，14（3）：5338-5366. doi：10.3390/ijms14035338 .
10	MINCIACCHI V R， FREEMAN M R， DI VIZIO D. Extracellular vesicles in cancer： Exosomes， microvesicles and the emerging role of large oncosomes[J]. Semin Cell Dev Biol，2015，40：41-51. doi：10.1016/j.semcdb.2015.02.010 .
11	SAHU R， KAUSHIK S， CLEMENT C C， et al. Microautophagy of cytosolic proteins by late endosomes[J]. Dev Cell，2011，20（1）：131-139. doi：10.1016/
	j.devcel.2010.12.003.
12	RECORD M. Intercellular communication by exosomes in placenta： A possible role in cell fusion?[J]. Placenta，2014，35（5）：297-302. doi：10.1016/j.placenta. 2014.02.009 .
13	YELLON D M， DAVIDSON S M. Exosomes： Nanoparticles involved in cardioprotection?[J]. Circ Res，2014，114（2）：325-332. doi：10.1161/CIRCRESA HA.113.300636 .
14	HENNE W M， BUCHKOVICH N J， EMR S D. The ESCRT pathway[J]. Dev Cell，2011，21（1）：77-91. doi：10.1016/j.devcel.2011.05.015 .
15	HURLEY J H. ESCRTs are everywhere[J]. EMBO J，2015，34（19）：2398-2407. doi：10.15252/embj.20159 2484 .
16	CASTRO B M， PRIETO M， SILVA L C. Ceramide： A simple sphingolipid with unique biophysical properties[J]. Prog Lipid Res，2014，54：53-67. doi：10.1016/j.plipres.2014.01.004 .
17	VAN NIEL G， D’ANGELO G， RAPOSO G. Shedding light on the cell biology of extracellular vesi-cles[J]. Nat Rev Mol Cell Biol，2018，19（4）：213-228. doi：10.1038/nrm.2017.125 .
18	ZHAO H， YANG L， BADDOUR J， et al. Tumor microenvironment derived exosomes pleiotropically modulate cancer cell metabolism[J]. Elife，2016，5：e10250. doi：10.7554/elife.10250 .
19	EITAN E， SUIRE C， ZHANG S， et al. Impact of lysosome status on extracellular vesicle content and release[J]. Ageing Res Rev，2016，32：65-74. doi：10 .
	1016/j.arr.2016.05.001.
20	CLAYTON A， HARRIS C L， COURT J， et al. Antigen-presenting cell exosomes are protected from complement-mediated lysis by expression of CD55 and CD59[J]. Eur J Immunol，2003，33（2）：522-531. doi：10.1002/immu.200310028 .
21	IMAI T， TAKAHASHI Y， NISHIKAWA M， et al. Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice[J]. J Extracell Vesicles，2015，4：26238. doi：10.3402/jev.v4.26238 .
22	VLASSOV A V， MAGDALENO S， SETTERQUIST R， et al. Exosomes： Current knowledge of their composition， biological functions， and diagnostic and therapeutic potentials[J]. Biochim Biophys Acta，2012，1820（7）：940-948. doi：10.1016/j.bbagen.2012.03.017 .
23	CASERTA S， GHEZZI P. Release of redox enzymes and micro-RNAs in extracellular vesicles， during infection and inflammation[J]. Free Radic Biol Med，2021，169：248-257. doi：10.1016/j.freeradbiomed.2021. 04.010 .
24	MITTELBRUNN M， GUTIÉRREZ-VÁZQUEZ C， VILLARROYA-BELTRI C， et al. Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells[J]. Nat Commun，2011，2：282. doi：10.1038/ncomms1285 .
25	VALADI H， EKSTRÖM K， BOSSIOS A， et al. Exosome-mediated transfer of mRNAs and micro-RNAs is a novel mechanism of genetic exchange between cells[J]. Nat Cell Biol，2007，9（6）：654-659. doi：10.1038/ncb1596 .
26	HUANG Z， XU A. Adipose extracellular vesicles in intercellular and inter-organ crosstalk in metabolic health and diseases[J]. Front Immunol，2021，12：608680. doi：10.3389/fimmu.2021.608680 .
27	FAN T， SUN N， HE J. Exosome-derived lncRNAs in lung cancer[J]. Front Oncol，2020，10：1728. doi：10.3389/fonc.2020.01728 .
28	HEWSON C， CAPRARO D， BURDACH J， et al. Extracellular vesicle associated long non-coding RNAs functionally enhance cell viability[J]. Non Coding RNA Res，2016，1（1）：3-11. doi：10.1016/j.ncrna.2016.06.001 .
29	ZHANG Y， LIU Y， LIU H， et al. Exosomes： Biogenesis， biologic function and clinical potential[J]. Cell Biosci，2019，9：19. doi：10.1186/s13578-019-0282-2 .
30	AYALA-MAR S， DONOSO-QUEZADA J， GALLO-VILLANUEVA R C， et al. Recent advances and challenges in the recovery and purification of cellular exosomes[J]. Electrophoresis，2019，40（23/24）：3036-3049. doi：10.1002/elps.201800526 .
31	LIVSHITS M A， KHOMYAKOVA E， EVTUSHEN-KO E G， et al. Isolation of exosomes by differential centrifugation： Theoretical analysis of a commonly used protocol[J]. Sci Rep，2015，5：17319. doi：
33	1038/srep17319.
32	LI P， KASLAN M， LEE S H， et al. Progress in exosome isolation techniques[J]. Theranostics，2017，7（3）：789-804. doi：10.7150/thno.18133 .
35	DEREGIBUS M C， FIGLIOLINI F， D’ANTICO S， et al. Charge-based precipitation of extracellular vesicles[J]. Int J Mol Med，2016，38（5）：1359-1366. doi：10.3892/ijmm.2016.2759 .
34	王耀杰，颜晰，赵立波，等. 外泌体分离和纯化方法研究进展[J].中华检验医学杂志，2021，44（2）：167-170. doi：10.3760/cma.j.cn114452-20200525-00497 .
	WANG Y J， YAN X， ZHAO L B， et al. Advances in methods for isolation and purification of exosomes[J]. Zhonghua Jianyan Yixue Zazhi，2021，44（2）：167-170.
37	GUAN S， YU H， YAN G， et al. Characterization of urinary exosomes purified with size exclusion chromatography and ultracentrifugation[J]. J Proteome Res，2020，19（6）：2217-2225. doi：10.1021/acs.jproteo me.9b00693 .
36	XU H， LIAO C， ZUO P， et al. Magnetic-based microfluidic device for on-chip isolation and detection of tumor-derived exosomes[J]. Anal Chem，2018，90（22）：13451-13458. doi：10.1021/acs.analchem.8b03 272 .
39	KANWAR S S， DUNLAY C J， SIMEONE D M， et al. Microfluidic device （ExoChip） for on-chip isolation， quantification and characterization of circulating exosomes[J]. Lab Chip，2014，14（11）：1891-1900. doi：10.1039/c4lc00136b .
38	MARTÍNEZ-GREENE J A， HERNÁNDEZ-ORTEGA K， QUIROZ-BAEZ R， et al. Quantitative proteomic analysis of extracellular vesicle subgroups isolated by an optimized method combining polymer-based precipitation and size exclusion chromatography[J]. J Extracell Vesicles，2021，10（6）：e12087. doi：10.1002/ jev2.12087 .
41	THÉRY C， WITWER K W， AIKAWA E， et al. Minimal information for studies of extracellular vesicles 2018 （MISEV2018）： A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines[J]. J Extracell Vesicles，2018，7（1）：1535750. doi：10.1080/20013078.2018.153575 .
40	SHAO H， IM H， CASTRO C M， et al. New technologies for analysis of extracellular vesicles[J]. Chem Rev，2018，118（4）：1917-1950. doi：10.1021/acs. chemrev.7b00534 .
43	WU Y， DENG W， KLINKE D J 2 ND. Exosomes： Improved methods to characterize their morphology， RNA content， and surface protein biomarkers[J]. Analyst，2015，140（19）：6631-6642. doi：10.1039/c5an
	00688k.
42	XU R， GREENING D W， ZHU H J， et al. Extracellular vesicle isolation and characterization： Toward clinical application[J]. J Clin Invest，2016，126（4）：1152-1162. doi：10.1172/JCI81129 .
45	POSPICHALOVA V， SVOBODA J， DAVE Z， et al. Simplified protocol for flow cytometry analysis of fluorescently labeled exosomes and microvesicles using dedicated flow cytometer[J]. J Extracell Vesi-cles，2015，4：25530. doi：10.3402/jev.v4.25530 .
44	ZHANG Y， BI J， HUANG J， et al. Exosome： A review of its classification， isolation techniques， sto-rage， diagnostic and targeted therapy applications[J]. Int J Nanomedicine，2020，15： 6917-6934. doi：10.2147/ IJN.S264498 .
47	SZATANEK R， BAJ-KRZYWORZEKA M， ZIMOCH J， et al. The methods of choice for extracellular vesicles （EVs） characterization[J]. Int J Mol Sci，2017，18（6）：1153. doi：10.3390/ijms18061153 .
46	MAAS S L N， DE VRIJ J， VAN DER VLIST E J， et al. Possibilities and limitations of current technologies for quantification of biological extracellular vesicles and synthetic mimics[J]. J Control Release，2015，200：87-96. doi：10.1016/j.jconrel.2014.12.041 .
49	HUANG X， YUAN T， TSCHANNEN M， et al. Characterization of human plasma-derived exosomal RNAs by deep sequencing[J]. BMC Genomics，2013，14：319. doi：10.1186/1471-2164-14-319 .
48	GONZALES P A， PISITKUN T， HOFFERT J D， et al. Large-scale proteomics and phosphoproteomics of urinary exosomes[J]. J Am Soc Nephrol，2009，20（2）：363-379. doi：10.1681/ASN.2008040406 .
51	BAHN J H， ZHANG Q， LI F， et al. The landscape of microRNA， Piwi-interacting RNA， and circular RNA in human saliva[J]. Clin Chem，2015，61（1）：221-230. doi：10.1373/clinchem.2014.230433 .
50	PARCHEM J， PAPANNA R， YANG S， et al. 89： Exploring the diagnostic and prognostic potential of amniotic fluid exosomes in twin-twin transfusion syndrome （TTTS）[J]. Am J Obstet Gynecol，2017，216（S1）：S63-S64. doi：10.1016/j.ajog.2016.11.977 .
53	YAGI Y， OHKUBO T， KAWAJI H， et al. Next-generation sequencing-based small RNA profiling of cerebrospinal fluid exosomes[J]. Neurosci Lett，2017，636：48-57. doi：10.1016/j.neulet.2016.10.042 .
52	WU C X， LIU Z F. Proteomic profiling of sweat exosome suggests its involvement in skin immuni-ty[J]. J Investig Dermatol，2018，138（1）：89-97. doi：10.1016/j.jid.2017.05.040 .
55	YANG C， GUO W B， ZHANG W S， et al. Comprehensive proteomics analysis of exosomes derived from human seminal plasma[J]. Andrology，2017，5（5）：1007-1015. doi：10.1111/andr.12412 .
54	HOSHINO A， KIM H S， BOJMAR L， et al. Extracellular vesicle and particle biomarkers define multiple human cancers[J]. Cell，2020，182（4）：1044-1061.e18. doi：10.1016/j.cell.2020.07.009 .
57	DUTTA S， KUMAR S， HYETT J， et al. Molecular targets of aspirin and prevention of preeclampsia and their potential association with circulating extracellular vesicles during pregnancy[J]. Int J Mol Sci，2019，20（18）：4370. doi：10.3390/ijms20184370 .
56	LU Q， ZHANG J， ALLISON R， et al. Identification of extracellular delta-catenin accumulation for prostate cancer detection[J]. Prostate，2009，69（4）：411-418. doi：10.1002/pros.20902 .
59	MOON P G， LEE J E， YOU S， et al. Proteomic analysis of urinary exosomes from patients of early IgA nephropathy and thin basement membrane nephropathy[J]. Proteomics，2011，11（12）：2459-2475. doi：10.1002/pmic.201000443 .
58	MACHIDA T， TOMOFUJI T， EKUNI D， et al. MicroRNAs in salivary exosome as potential biomarkers of aging[J]. Int J Mol Sci，2015，16（9）：21294-21309. doi：10.3390/ijms160921294 .
61	董梦茹. 重度烧伤休克期大鼠血清外泌体的提取鉴定及miRNA表达谱分析[D].广州：南方医科大学，2020.
	DONG M R. Extraction and identification of serum exosomes and analysis of miRNA expression profiles in severe burns rats during shock stage[D]. Guangzhou： Southern Medical University，2020.
60	SAMANTA L， PARIDA R， DIAS T R， et al. The enigmatic seminal plasma： A proteomics insight from ejaculation to fertilization[J]. Reprod Biol Endocrinol，2018，16（1）：41. doi：10.1186/s12958-018-0358-6 .
63	马晓楠，路陆，黄琰潼，等. 外泌体microRNA在心血管疾病中的研究进展及其法医学应用前景[J].法医学杂志，2022，38（2）：258-262. doi：10.12116/j.issn.1004-5619.2020.400911 .
	MA X N， LU L， HUANG Y T， et al. Research progress of exosomal microRNA in cardiovascular disease and its forensic application prospects[J]. Fayi-xue Zazhi，2022，38（2）：258-262.
62	HUANG Y， CHENG L， TURCHINOVICH A， et al. Influence of species and processing parameters on recovery and content of brain tissue-derived extracellular vesicles[J]. J Extracell Vesicles，2020，9（1）：1785746. doi：10.1080/20013078.2020.1785746 .
65	HARISCHANDRA D S， GHAISAS S， ROKAD D， et al. Environmental neurotoxicant manganese regulates exosome-mediated extracellular miRNAs in cell culture model of Parkinson’s disease： Relevance to α-synuclein misfolding in metal neurotoxicity[J]. Neurotoxicology，2018，64：267-277. doi：10.1016/j.neuro. 2017.04.007 .
64	OGAWA Y， KANAI-AZUMA M， AKIMOTO Y， et al. Exosome-like vesicles in Gloydius blomhoffii blomhoffii venom[J]. Toxicon，2008，51（6）：984-993. doi：10.1016/j.toxicon.2008.02.003 .
67	SOUZA-IMBERG A， CARNEIRO S M， GIAN-NOTTI K C， et al. Origin and characterization of small membranous vesicles present in the venom of Crotalus durissus terrificus [J]. Toxicon，2017，136：27-33. doi：10.1016/j.toxicon.2017.06.013 .
66	CHO Y E， MEZEY E， HARDWICK J P， et al. Increased ethanol-inducible cytochrome P450-2E1 and cytochrome P450 isoforms in exosomes of alcohol-exposed rodents and patients with alcoholism through oxidative and endoplasmic reticulum stress[J]. Hepatol Commun，2017，1（7）：675-690. doi：10.1002/hep4. 1066 .
69	WANG X， SUN L， ZHOU Y， et al. Heroin abuse and/or HIV infection dysregulate plasma exosomal miRNAs[J]. J Neuroimmune Pharmacol，2020，15（3）：400-408. doi：10.1007/s11481-019-09892-9 .
68	孟运乐. 甲基苯丙胺促使α-突触核蛋白传递至星形胶质细胞的相关病理损伤机制研究[D].广州：南方医科大学，2020.
	MENG Y L. Mechanism of pathological damage in astrocytes after alpha-synuclein transferred to astrocytes promoted by METH[D]. Guangzhou： Southern Medical University，2020.
71	郜峥翔，罗奇志，张亮，等. 大鼠血液外泌体中外源性γ-羟基丁酸的检测[J].法医学杂志，2022，38（2）：212-216. doi：10.12116/j.issn.1004-5619.2021.410116 .
	GAO Z X， LUO Q Z， ZHANG L， et al. Detection of exogenous γ-hydroxybutyric acid in rat blood exosomes[J]. Fayixue Zazhi，2022，38（2）：212-216.
70	JANAS A M， SAPOŃ K， JANAS T， et al. Exosomes and other extracellular vesicles in neural cells and neurodegenerative diseases[J]. Biochim Biophys Acta，2016，1858（6）：1139-1151. doi：10.1016/j.bbam em.2016.02.011 .
73	TIAN F， SHEN Y， CHEN Z， et al. No significant difference between plasma miRNAs and plasma-derived exosomal miRNAs from healthy people[J]. Biomed Res Int，2017，2017：1304816. doi：10.1155/2017/1304816 .
72	GRUZDEV S K， YAKOVLEV A A， DRUZHKOVA T A， et al. The missing link： How exosomes and miRNAs can help in bridging psychiatry and molecular biology in the context of depression， bipolar disorder and schizophrenia[J]. Cell Mol Neurobiol，2019，39（6）：729-750. doi：10.1007/s10571-019-00684-6 .
75	LIU J， SU B. Integrated analysis supports ATXN1 as a schizophrenia risk gene[J]. Schizophr Res，2018，195：298-305. doi：10.1016/j.schres.2017.10.010 .
74	RAISZADEH M M， ROSS M M， RUSSO P S， et al. Proteomic analysis of eccrine sweat： Implications for the discovery of schizophrenia biomarker proteins[J]. J Proteome Res，2012，11（4）：2127-2139. doi：10.1021/pr2007957 .
77	TSILIONI I， THEOHARIDES T C. Extracellular vesicles are increased in the serum of children with autism spectrum disorder， contain mitochondrial DNA， and stimulate human microglia to secrete IL-1β[J]. J Neuroinflammation，2018，15（1）：239. doi：10.1186/s12974-018-1275-5 .

方法	优点	缺点
离心法
差速离心法^[34]	操作简单	操作用时较长，设备价格高昂，分离纯度低
密度梯度离心法	分离纯度高，理化性质保留^[34]	操作复杂，分离效率低^[32]
沉淀法
聚合物沉淀法	操作简单^[34]	纯度和回收率低^[33]
有机溶剂沉淀法^[34]	廉价、操作简单、省时，理化性质保留	操作复杂
粒径分离法
超滤法^[34]	设备廉价	过滤膜的堵塞会降低回收率
排阻色谱法	理化性质保留，避免外泌体聚集，分离纯度高于离心法^[35]	操作用时较长^[34]
免疫亲和法（磁分离法）	特异性高，分离效率高^[32]	反应试剂昂贵^[34]
微流控技术	省时，有高通量潜力^[32]	分离效率取决于装置原理^[36]

方法	优点	缺点
离心法
差速离心法^[34]	操作简单	操作用时较长，设备价格高昂，分离纯度低
密度梯度离心法	分离纯度高，理化性质保留^[34]	操作复杂，分离效率低^[32]
沉淀法
聚合物沉淀法	操作简单^[34]	纯度和回收率低^[33]
有机溶剂沉淀法^[34]	廉价、操作简单、省时，理化性质保留	操作复杂
粒径分离法
超滤法^[34]	设备廉价	过滤膜的堵塞会降低回收率
排阻色谱法	理化性质保留，避免外泌体聚集，分离纯度高于离心法^[35]	操作用时较长^[34]
免疫亲和法（磁分离法）	特异性高，分离效率高^[32]	反应试剂昂贵^[34]
微流控技术	省时，有高通量潜力^[32]	分离效率取决于装置原理^[36]

方法	检测内容	优点	缺点
动态光散射技术^[42]	大小	测定下限低，适用于单分散体系的测定	不适合测量大尺寸的外泌体样本，对多分散样本不准确
纳米颗粒跟踪分析技术	大小和体积分数	省时，可以实时观察外泌体；分辨率高于流式细胞仪，测量的直径下限可达30 nm^[42，45]	操作复杂，很难从外泌体中分辨出被污染的蛋白质；摄像水平和检测阈值将影响外泌体的定量^[46]

方法	检测内容	优点	缺点
动态光散射技术^[42]	大小	测定下限低，适用于单分散体系的测定	不适合测量大尺寸的外泌体样本，对多分散样本不准确
纳米颗粒跟踪分析技术	大小和体积分数	省时，可以实时观察外泌体；分辨率高于流式细胞仪，测量的直径下限可达30 nm^[42，45]	操作复杂，很难从外泌体中分辨出被污染的蛋白质；摄像水平和检测阈值将影响外泌体的定量^[46]

[1]	Wen LI, Hao-zhe LI, Chen CHEN, Wei-xiong CAI. Research Progress and Application Prospect of Facial Micro-Expression Analysis in Forensic Psychiatry [J]. Journal of Forensic Medicine, 2023, 39(5): 493-500.
[2]	Zhong-hua WANG, Shu-jin LI. Research Progress on Molecular Biology of Human Height Estimation [J]. Journal of Forensic Medicine, 2023, 39(5): 487-492.
[3]	Lu CHEN, Zhe ZHOU, Sheng-qi WANG. Process of Forensic Medicine in DNA Identification of Aged Human Remains [J]. Journal of Forensic Medicine, 2023, 39(5): 478-486.
[4]	Yong ZENG, Dong-hua ZOU, Ying FAN, Qing XU, Lu-yang TAO, Yi-jiu CHEN, Zheng-dong LI. Research Progress and Forensic Application of Human Vascular Finite Element Modeling and Biomechanics [J]. Journal of Forensic Medicine, 2023, 39(5): 471-477.
[5]	Hong-xia HAO, Jie-min CHEN, Rong-rong WANG, Xiao-ying YU, Meng WANG, Zhi-lu ZHOU, Yan-liang SHENG, Wen-tao XIA. The Value of VR-PVEP in Objective Evaluation of Monocular Refractive Visual Impairment [J]. Journal of Forensic Medicine, 2023, 39(4): 382-387.
[6]	Jie BAI, Jing SUN, Xiao-guang CHENG, Fan LIU, Hua LIU, Xu WANG. Construction and Application of Rib Fracture Diagnosis Model Based on YOLOv3 Algorithm [J]. Journal of Forensic Medicine, 2023, 39(4): 343-349.
[7]	Fei FAN, Juan WU, Zhen-hua DENG. Application Progress of Objective Audiological Detection Techniques in Forensic Clinical Medicine [J]. Journal of Forensic Medicine, 2023, 39(4): 360-366.
[8]	Jian XIANG, Xu WANG, Li-li YU, Kang-jia JIN, Ying-kai YANG. Objective Assessment of Visual Field Defects Caused by Optic Chiasm and Its Posterior Visual Pathway Injury [J]. Journal of Forensic Medicine, 2023, 39(4): 350-359.
[9]	Qing-qing XIANG, Li-fang CHEN, Qin SU, Yu-kun DU, Pei-yan LIANG, Xiao-dong KANG, He SHI, Qu-yi XU, Jian ZHAO, Chao LIU, Xiao-hui CHEN. Research Progress on Microbial Community Succession in the Postmortem Interval Estimation [J]. Journal of Forensic Medicine, 2023, 39(4): 399-405.
[10]	Zhang-ming GAO, Jing-yu SHI, Hao ZENG, Xue-jun ZHANG. Rapid Determination of Bucinnazine in Blood by UPLC-MS/MS [J]. Journal of Forensic Medicine, 2023, 39(4): 388-392.
[11]	Yu-qi CAO, Yan SHI, Ping XIANG, Yin-long GUO. Research Progress on Machine Learning Assisted Non-Targeted Screening Strategy for Identification of Fentanyl Analogs [J]. Journal of Forensic Medicine, 2023, 39(4): 406-416.
[12]	Xu WANG. Legal Theories， Disability Models and Principles of Disability Assessment [J]. Journal of Forensic Medicine, 2023, 39(4): 329-336.
[13]	Ran LI, Hong-yu SUN. Methods and Research Hotspots of Forensic Kinship Testing [J]. Journal of Forensic Medicine, 2023, 39(3): 231-239.
[14]	Xiao-yan MA, Hong-yu SUN, Qing LI. Research Progresses of Tri-Allelic Patterns in Autosomal STR in Forensic DNA Analysis [J]. Journal of Forensic Medicine, 2023, 39(3): 240-246.
[15]	Hao-yang WANG, Jian WU, Qian ZHANG, Xin-feng MIN, Xiu-yan LIU, Yin-long GUO. Structural Analysis and Characterization of 4-F-α-PVP Analog 4-F-3-Methyl-α- PVP Hydrochloride [J]. Journal of Forensic Medicine, 2023, 39(2): 144-150.