1.Identification of Human Body Fluid Stains and Non-Biological Stains by Three-Dimensional Fluorescence Spectroscopy.
Jing-Jing CHANG ; Hui ZHOU ; Jin ZHANG ; Xiao-Yu XU ; Feng WANG ; Sheng-Jun XIONG ; Guang-Feng ZHANG ; Xue-Ying YANG ; Kai-Hui LIU
Journal of Forensic Medicine 2023;39(1):40-44
OBJECTIVES:
To establish a rapid and nondestructive identification method for human body fluid stains and non-biological stains using three-dimensional fluorescence spectroscopy.
METHODS:
The collected three-dimensional fluorescence spectrum data of human saliva, 3% blood, coffee and Fanta® stains were processed with dimensionality reduction. After wavelet transform, spectral denoising and feature extraction, the classification formula was established. The Fisher discriminant was used for spectrum matching and recognition to establish the analysis method to distinguish stain types.
RESULTS:
According to the results of data training and comparison, all the recognition accuracies of Fanta®, coffee, saliva and blood were more than 91.39%. Among them, saliva reached 100% recognition accuracy.
CONCLUSIONS
Three-dimensional fluorescence spectroscopy is a potential method for rapid and nondestructive identification of biological and non-biological stains.
Humans
;
Forensic Medicine/methods*
;
Coloring Agents/analysis*
;
Coffee
;
Spectrometry, Fluorescence
;
Body Fluids/chemistry*
2.Roles of exosome-derived non-coding RNA in tumor micro-environment and its clinical application.
Qinyi DOU ; Jiazheng WANG ; Yingshuo YANG ; Wei ZHUO
Journal of Zhejiang University. Medical sciences 2023;52(4):429-438
Tumor-derived exosomes play an important role in the tumor micro-environment. The exosome-derived non-coding RNAs are transmitted in the tumor microenvironment in three ways, communication between tumor cells, normal cells affecting tumor cells, and tumor cells affecting normal cells. Through these three ways, exosomal non-coding RNAs are involved in the regulation of tumor progression, affecting tumor angiogenesis, tumor invasiveness, drug resistance, stemness, tumor metabolic repro-gramming and immune escape, resulting in dual roles in promoting or inhibiting tumor development. Exosomes have a membranous structure and their contents are resistant to degradation by extracellular proteases and remain highly stable in body fluids, thus exosome-derived non-coding RNAs are expected to serve as diagnostic and prognostic indicators for a variety of cancers. In addition, exosomes can be used to deliver non-coding RNAs for targeted therapy, or to knock down or modify tumor-promoting non-coding RNAs for tumor therapy. This article reviews the function and communication mechanism of exosomal non-coding RNAs in the tumor microenvironment, including their pathways of action, effects, potential values for tumor biomarkers and treatment targets. This article also points out the issues that need to be further studied in order to promote the progress of extracellular non-coding RNAs in cancer research and their application in tumor diagnosis and treatment.
Humans
;
Exosomes
;
Neoplasms/genetics*
;
Biomarkers, Tumor
;
Body Fluids
;
RNA, Untranslated/genetics*
;
Tumor Microenvironment
3.Research Progress on Biological Matrix Reference Materials in Forensic Toxicology.
Hang CHEN ; Jing HU ; Zheng QIAO ; Hong-Xiao DENG ; Min LÜ ; Wei LIU
Journal of Forensic Medicine 2023;39(2):176-185
Biological matrix reference material is a reference material that combines the target material with the biological matrix. The biological matrix reference material has higher consistency with the authentic specimens in forensic toxicology, and its application has a positive effect on improving the accuracy of test results. This paper reviews the research on the matrix reference materials corresponding to three common biological test materials (blood, urine and hair). In order to provide reference for the development and application of biological matrix reference materials in forensic toxicology, this paper mainly introduces the research progress of preparation technology of biological matrix reference materials and some existing products and their parameters evaluation.
Forensic Toxicology/methods*
;
Hair
;
Body Fluids
4.Construction of an Analysis Model of mRNA Markers in Menstrual Blood Based on Naïve Bayes and Multivariate Logistic Regression Methods.
Qi ZHANG ; He-Miao ZHAO ; Kang YANG ; Jing CHEN ; Rui-Qin YANG ; Chong WANG
Journal of Forensic Medicine 2023;39(5):447-451
OBJECTIVES:
To establish the menstrual blood identification model based on Naïve Bayes and multivariate logistic regression methods by using specific mRNA markers in menstrual blood detection technology combined with statistical methods, and to quantitatively distinguish menstrual blood from other body fluids.
METHODS:
Body fluids including 86 menstrual blood, 48 peripheral blood, 48 vaginal secretions, 24 semen and 24 saliva samples were collected. RNA of the samples was extracted and cDNA was obtained by reverse transcription. Five menstrual blood-specific markers including members of the matrix metalloproteinase (MMP) family MMP3, MMP7, MMP11, progestogens associated endometrial protein (PAEP) and stanniocalcin-1 (STC1) were amplified and analyzed by electrophoresis. The results were analyzed by Naïve Bayes and multivariate logistic regression.
RESULTS:
The accuracy of the classification model constructed was 88.37% by Naïve Bayes and 91.86% by multivariate logistic regression. In non-menstrual blood samples, the distinguishing accuracy of peripheral blood, saliva and semen was generally higher than 90%, while the distinguishing accuracy of vaginal secretions was lower, which were 16.67% and 33.33%, respectively.
CONCLUSIONS
The mRNA detection technology combined with statistical methods can be used to establish a classification and discrimination model for menstrual blood, which can distignuish the menstrual blood and other body fluids, and quantitative description of analysis results, which has a certain application value in body fluid stain identification.
Female
;
Humans
;
RNA, Messenger/metabolism*
;
Bayes Theorem
;
Logistic Models
;
Menstruation
;
Body Fluids
;
Saliva
;
Semen
;
Forensic Genetics/methods*
5.Application of SNaPshot Technology in Semen-Specific cSNP Genetic Marker.
Rui-Yang TAO ; Shou-Yu WANG ; Chun-Yan YUAN ; Ruo-Cheng XIA ; Cheng-Tao LI
Journal of Forensic Medicine 2023;39(5):465-470
OBJECTIVES:
To explore the feasibility of genetic marker detection of semen-specific coding region single nucleotide polymorphism (cSNP) based on SNaPshot technology in semen stains and mixed body fluid identification.
METHODS:
Genomic DNA (gDNA) and total RNA were extracted from 16 semen stains and 11 mixtures composed of semen and venous blood, and the total RNA was reverse transcribed into complementary DNA (cDNA). The cSNP genetic markers were screened on the validated semen-specific mRNA coding genes. The cSNP multiplex detection system based on SNaPshot technology was established, and samples were genotyped by capillary electrophoresis (CE).
RESULTS:
A multiplex detection system containing 5 semen-specific cSNPs was successfully established. In 16 semen samples, except the cSNP located in the TGM4 gene showed allele loss in cDNA detection results, the gDNA and cDNA typing results of other cSNPs were highly consistent. When detecting semen-venous blood mixtures, the results of cSNP typing detected were consistent with the genotype of semen donor and were not interfered by the genotype of venous blood donor.
CONCLUSIONS
The method of semen-specific cSNPs detection by SNaPshot technology method can be applied to the genotyping of semen (stains) and provide information for determining the origin of semen in mixed body fluids (stains).
Genetic Markers
;
Semen
;
Polymorphism, Single Nucleotide
;
DNA, Complementary/genetics*
;
Body Fluids
;
RNA, Messenger/genetics*
;
DNA
;
Saliva
;
Forensic Genetics/methods*
6.Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification.
Sun-Xiang WEI ; Hui-Xiang CHEN ; Sheng HU ; Yi-Xia ZHAO ; Hui-Xia SHI ; Zhe WANG ; Wen LI ; An-Quan JI ; Qi-Fan SUN
Journal of Forensic Medicine 2022;38(6):719-725
OBJECTIVES:
To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.
METHODS:
The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.
RESULTS:
There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/μL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/μL, and the discrimination accuracy was 100%.
CONCLUSIONS
In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.
Female
;
Humans
;
Body Fluids/chemistry*
;
MicroRNAs/analysis*
;
Real-Time Polymerase Chain Reaction/methods*
;
Saliva/chemistry*
;
Semen/chemistry*
;
Male
7.Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification.
Shou-Yu WANG ; Rui-Yang TAO ; Yi-Ping HOU ; Cheng-Tao LI
Journal of Forensic Medicine 2022;38(6):763-773
In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.
Forensic Medicine/methods*
;
Body Fluids/chemistry*
;
RNA/analysis*
;
Feces
;
Forensic Genetics
;
Semen/chemistry*
;
Saliva/chemistry*

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