Comparison of Application of MD-VF-Auto SEM Method and Plankton Gene Multiplex PCR System in the Diagnosis of Drowning / 法医学杂志

OBJECTIVES@#To compare the application effect of microwave digestion - vacuum filtration - automated scanning electron microscopy (MD-VF-Auto SEM) method and plankton gene multiplex PCR system in the diagnosis of drowning.@*METHODS@#Lung, liver and kidney tissue of 10 non-drowning cases and 50 drowning cases were prepared for further MD-VF-Auto SEM method analysis and plankton gene multiplex PCR system analysis. The positive detection rate of the two methods in each tissue was calculated.@*RESULTS@#The positive rate of the MD-VF-Auto SEM method detecting diatoms in drowning cases was 100%, and few diatoms were detected in the liver and kidney tissues of 6 non-drowning cases. By using the plankton gene multiplex PCR system, the diatom positive rate of drowning cases was 84%, and all the non-drowning cases were negative. There were significant differences in the positive rate of the liver, kidney tissues between MD-VF-Auto SEM method and plankton gene multiplex PCR system (P<0.05), as well as the total positive rate of cases. However, no significant differences were found in the positive rates of lung tissues (P>0.05).@*CONCLUSIONS@#MD-VF-Auto SEM method is more sensitive than plankton gene multiplex PCR system in diatom test. But the plankton gene multiplex PCR system can also detect plankton other than diatoms. Combination of the two methods can provide a more reliable basis for the diagnosis of drowning.

Application of Diatoms Quantitative Analysis in the Diagnosis of Drowning / 法医学杂志

OBJECTIVES@#To retrospectively analyze diatom test cases of corpses in water and discuss the value of quantitative analysis of diatoms in the diagnosis of drowning.@*METHODS@#A total of 490 cases of water-related death were collected. They were divided into drowning group and postmortem immersion group according to the cause of death. Diatoms in lung, liver, kidney tissue and water sample were analyzed quantitatively by microwave digestion-vacuum filtration-automated scanning electron microscopy (MD-VF-Auto SEM) method. The ratios of content of diatoms in lung tissue and water sample (CL/CD) were calculated.@*RESULTS@#The results of diatom test for three organs (lung, liver and kidney) were all positive in 400 cases (85.5%); the content of diatom in lung, liver, kidney tissues, and water samples of drowning group were (113 235.9±317 868.1), (26.7±75.6), (23.3±52.2) and (12 113.3±21 760.0) cells/10 g, respectively; the species of diatom were (7.5±2.8), (2.6±1.9), (2.9±2.1) and (8.9±3.0) types, respectively; the CL/CD of drowning group and postmortem immersion group were (100.6±830.7) and (0.3±0.4), respectively.@*CONCLUSIONS@#Quantitative analysis of diatoms can provide supportive evidence for the diagnosis of drowning, and the parameter CL/CD can be introduced into the analysis to make a more accurate diagnosis of drowning.

Bibliometrics Analysis in English and Chinese Literature on Drowning in Forensic Medicine from 1991 to 2020 / 法医学杂志

OBJECTIVES@#To explore the research hotspots and development trends of the field of forensic drowning from 1991 to 2020 by bibliometrics methods.@*METHODS@#Based on Web of Science, CNKI database, Wanfang Data knowledge service platform, python 3.9.2, CiteSpace 5.8.R3, Gephi 0.9.2, etc. were used to analyze the publishing trends, countries/regions, institutions, authors and topics of the study on drowning.@*RESULTS@#A total of 631 English literature were obtained, including 59 articles from Chinese authors, and 386 Chinese literature were obtained. The Chinese and English journals with the largest number of related literatures were Chinese Journal of Forensic Science (80 articles) and Forensic Science International (106 articles), respectively. Japan published the most articles in English, and China ranked third. Osaka City Univ (Japan, 28 articles) published the most English articles, and Guangzhou Forens Sci Inst (China, 22 articles) ranked second. Among Chinese literature, Guangzhou Forens Sci Inst (32 articles) published the most. The topic analysis of Chinese and English literature showed that diatom examination, virtual autopsy, postmortem biochemical examination, the nature of death, and postmortem submersion interval were the hot spots of current research, but English literature had more studies on new technologies and methods, while Chinese literature was more inclined to practice, application and experience summary.@*CONCLUSIONS@#The number of literature in forensic medicine on drowning is relatively stable. The scope of international and domestic collaborations in this field is still limited. The automated examination of diatoms, the establishment of diatom DNA barcodes and virtual autopsy will be the most important research hotspots in the coming period and are expected to achieve breakthroughs in drowning diagnosis, drowning location inference, postmortem submersion interval estimation, etc.

Change Rule of Diatom in Zhangweixin River in Dezhou / 法医学杂志

OBJECTIVES@#To study the annual variation of diatoms in Zhangweixin River, to provide theoretical support by using diatom examination to estimate the time and place of the corpse entering water, and to establish a diatom database.@*METHODS@#Samples were taken from 4 sampling sites in Decheng section of Zhangweixin River for 12 consecutive months. Non-metric multi-dimensional scaling (NMDS) analysis was performed on the species and content of diatom samples. Based on the sampling site of Tianqu Road, Sprensen similarity coefficient analysis was conducted with the data of other 3 sites in Decheng section and Leling section of Zhangweixin River and Ningjin section in previous studies.@*RESULTS@#The number of diatom species was positively correlated with diatom content. The average diatom content in different months ranged from 1 054 to 13 041/10 mL, and the species composition ranged from 8 to 16, with differences in dominant species. The similarity coefficient of diatom species within 2 km were all higher than 0.956 52, which could not be distinguished effectively. The similarity coefficients of Leling section and Ningjin section were 0.736 84 and 0.588 24 respectively, which could be effectively distinguished.@*CONCLUSIONS@#The species and content of diatom vary in different months in Zhangweixin River, and the composition of diatom species is different in different basins, which can provide reference for estimating the time and place of the corpse entering water in the river.

Construction and Application of YOLOv3-Based Diatom Identification Model of Scanning Electron Microscope Images / 法医学杂志

OBJECTIVES@#To construct a YOLOv3-based model for diatom identification in scanning electron microscope images, explore the application performance in practical cases and discuss the advantages of this model.@*METHODS@#A total of 25 000 scanning electron microscopy images were collected at 1 500× as an initial image set, and input into the YOLOv3 network to train the identification model after experts' annotation and image processing. Diatom scanning electron microscopy images of lung, liver and kidney tissues taken from 8 drowning cases were identified by this model under the threshold of 0.4, 0.6 and 0.8 respectively, and were also identified by experts manually. The application performance of this model was evaluated through the recognition speed, recall rate and precision rate.@*RESULTS@#The mean average precision of the model in the validation set and test set was 94.8% and 94.3%, respectively, and the average recall rate was 81.2% and 81.5%, respectively. The recognition speed of the model is more than 9 times faster than that of manual recognition. Under the threshold of 0.4, the mean recall rate and precision rate of diatoms in lung tissues were 89.6% and 87.8%, respectively. The overall recall rate in liver and kidney tissues was 100% and the precision rate was less than 5%. As the threshold increased, the recall rate in all tissues decreased and the precision rate increased. The F1 score of the model in lung tissues decreased with the increase of threshold, while the F1 score in liver and kidney tissues with the increase of threshold.@*CONCLUSIONS@#The YOLOv3-based diatom electron microscope images automatic identification model works at a rapid speed and shows high recall rates in all tissues and high precision rates in lung tissues under an appropriate threshold. The identification model greatly reduces the workload of manual recognition, and has a good application prospect.

Evaluation of Inspection Efficiency of Diatom Artificial Intelligence Search System Based on Scanning Electron Microscope / 法医学杂志

OBJECTIVES@#To explore the application values of diatom artificial intelligence (AI) search system in the diagnosis of drowning.@*METHODS@#The liver and kidney tissues of 12 drowned corpses were taken and were performed with the diatom test, the view images were obtained by scanning electron microscopy (SEM). Diatom detection and forensic expert manual identification were carried out under the thresholds of 0.5, 0.7 and 0.9 of the diatom AI search system, respectively. Diatom recall rate, precision rate and image exclusion rate were used to detect and compare the efficiency of diatom AI search system.@*RESULTS@#There was no statistical difference between the number of diatoms detected in the target marked by the diatom AI search system and the number of diatoms identified manually (P>0.05); the recall rates of the diatom AI search system were statistically different under different thresholds (P<0.05); the precision rates of the diatom AI system were statistically different under different thresholds(P<0.05), and the highest precision rate was 53.15%; the image exclusion rates of the diatom AI search system were statistically different under different thresholds (P<0.05), and the highest image exclusion rate was 99.72%. For the same sample, the time taken by the diatom AI search system to identify diatoms was only 1/7 of that of manual identification.@*CONCLUSIONS@#Diatom AI search system has a good application prospect in drowning cases. Its automatic diatom search ability is equal to that of experienced forensic experts, and it can greatly reduce the workload of manual observation of images.

Application Progress of High-Throughput Sequencing Technology in Forensic Diatom Detection / 法医学杂志

Diatom detection is an important method for identifying drowning and throwing corpses after death and inferring the drowning sites in forensic examination of corpses in water. In recent years,high-throughput sequencing technology has achieved rapid development and has been widely used in research related to diatom taxonomic investigations. This paper reviews the research status and prospects of high-throughput sequencing technology and its application in forensic diatom detection.

Research Progress of Automatic Diatom Test by Artificial Intelligence / 法医学杂志

Diatom test is the main laboratory test method in the diagnosis of drowning in forensic medicine. It plays an important role in differentiating the antemortem drowning from the postmortem drowning and inferring drowning site. Artificial intelligence (AI) automatic diatom test is a technological innovation in forensic drowning diagnosis which is based on morphological characteristics of diatom, the application of AI algorithm to automatic identification and classification of diatom in tissues and organs. This paper discusses the morphological diatom test methods and reviews the research progress of automatic diatom recognition and classification involving AI algorithms. AI deep learning algorithm can assist diatom testing to obtain objective, accurate, and efficient qualitative and quantitative analysis results, which is expected to become a new direction of diatom testing research in the drowning of forensic medicine in the future.

Review and Prospect of Diagnosis of Drowning Deaths in Water / 法医学杂志

Drowning is the death caused by asphyxiation due to fluid blocking the airway. In the practice of forensic medicine, it is the key to determine whether the corpse was drowned or entered the water after death. At the same time, the drowning site inference and postmortem submersion interval (PMSI) play an important role in the investigating the identity of the deceased, narrowing the investigation scope, and solving the case. Based on diatoms testing, molecular biology, imaging and artificial intelligence and other technologies, domestic and foreign forensic scientists have done relative research in the identification of the cause of death, drowning site inference and PMSI, and achieved certain results in forensic medicine application. In order to provide a reference for future study of bodies in the water, this paper summarizes the above research contents.

Application of Chlorophyte ChlB Gene and Cyanophyte NIES Gene in the Detection of Drowning-Related Plankton / 法医学杂志

Objective To construct a polymerase chain reaction-capillary electrophoresis （PCR-CE） detection method using ChlB gene and NIES gene, investigate the method's specificity and sensitivity, and to evaluate its application value in drowning diagnosis. Methods The specific primers ChlB and NIES were designed for the conserved sequence of chlorophyte ChlB gene and cyanophyte NIES gene in GenBank to construct PCR-CE detection method; 50 species of standard DNA samples were amplified; the sensitivity was determined by gradient concentration detection of positive standard samples; 25 actual cadaver lung tissue samples （drowned： 20, natural death： 5） were detected, and the simultaneous detection results of microwave digestion-vacuum filtration-automated scanning electron microscopy （MD-VF-Auto SEM） were simultaneously compared. Results The minimum DNA detection concentration of primers ChlB and NIES was 0.161 ng and 0.109 ng, respectively, which could specifically amplify chlorophyte （Chlorella pyrenoidosa） and cyanophyte ［Microcystis aeruginosa （producing and not producing toxin）］ widespread in water. The product fragments were 156 bp and 182 bp, respectively. The results of non-drowning tissues were negative. Conclusion This method has high sensitivity and specificity. It can be applied to the detection of plankton related to drowning and combined with MD-VF-Auto SEM method, can increase the detection range of plankton related to drowning and improve the evidence power of drowning diagnosis.

Application of Quantitative Analysis of Diatoms in Lung Tissue for the Diagnosis of Drowning of Experimental Animals / 法医学杂志

Objective To discuss the application value of diatom examination in lung tissue for the forensic diagnosis of drowning. Methods The experimental animals were divided randomly into drowning, postmortem submergence and dying on land group. Diatoms in lung tissue and drowning fluid were analyzed quantitatively by microwave digestion-vacuum filtration-automated scanning electron microscopy diatom examination method. The ratios of content of diatoms in lung tissue and drowning fluid （CL/CD ratio） were recorded. Results The CL/CD ratios of experimental rabbits in the drowning group （5.82±3.50） were much higher than that of postmortem submergence group （0.47±0.35）; the CL/CD ratios of different parts of the lung lobes of experimental pigs in the drowning group were higher than that of postmortem submergence group （P<0.05）; in seawater, brackish water, river fresh water and lake fresh water, the CL/CD ratios of experimental pigs in the drowning group were higher than that of postmortem submergence group （P<0.05）. In animal experiments, all the cases with CL/CD ratio >1.6 were from drowning group. Conclusion CL/CD ratio is an indicator with good application prospects in the diagnosis of drowning.

Application of Specific Fragment Length Polymorphism of Algae rDNA in Identification of Drowning Cases / 法医学杂志

OBJECTIVES@#To identify the drop-off location of victims in drowning cases, and confirm whether it is a fatal drowning or the victim is thrown into the water after death by detecting part of 5.8S sequence and second internal transcribed spacer （ITS2） （5.8S+ITS2） of diatom rDNA in water and organs.@*METHODS@#Two cases identified by diatom examination, which received by Nanjing Municipal Public Security Bureau Forensic Center, were taken as the research objects. The difference of the population structure of algae in water and human tissue was analysed by length polymorphism of 5.8S+ITS2 marker.@*RESULTS@#In case 1, similar species of diatom were detected from victim's lung and liver tissues and the water sample. Two kinds of DNA fragments with length of 330 bp and 376 bp were detected from victim's lung tissue and the water sample using 5.8S+ITS2 marker, which could confirm the victim was drowning before death. In case 2, there was no diatom found in victim's lung and liver tissues. Only one kind of DNA fragment with length of 331 bp and low relative fluorescence unit （RFU） was obtained from victim's lung tissue using 5.8S+ITS2 marker, thus the victim was thrown into the water after death.@*CONCLUSIONS@#The experimental results of the two cases in present study are consistent with the actual facts and the result of the diatom microscopic examination. The difference of population structure of specific microorganism in water and human tissue can be detected by 5.8S+ITS2 marker, which can help to identify the drop-off location of victims in drowning cases, and confirm whether it is a fatal drowning or the victim is thrown into the water after death.

Progress on Diatom Test in Drowning Cases / 法医学杂志

In drowning cases, it is difficult to judge whether the deceased died from drowning or throwing into the water after killed or identificate the cause of death of the decomposed corps in forensic practices. The diatom test is still considered as the important assistant evidence in drowning cases. This paper reviews research progress on technologies in recent years of forensic diatom test, and the application value of the new developing approach in the field of forensic medicine.

Comparative analysis between diatom nitric acid digestion method and plankton 16S rDNA PCR method / 法医学杂志

OBJECTIVE@#To compare and explore the application value of diatom nitric acid digestion method and plankton 16S rDNA PCR method for drowning identification.@*METHODS@#Forty drowning cases from 2010 to 2011 were collected from Department of Forensic Medicine of Wenzhou Medical University. Samples including lung, kidney, liver and field water from each case were tested with diatom nitric acid digestion method and plankton 16S rDNA PCR method, respectively. The Diatom nitric acid digestion method and plankton 16S rDNA PCR method required 20 g and 2 g of each organ, and 15 mL and 1.5 mL of field water, respectively. The inspection time and detection rate were compared between the two methods.@*RESULTS@#Diatom nitric acid digestion method mainly detected two species of diatoms, Centriae and Pennatae, while plankton 16S rDNA PCR method amplified a length of 162 bp band. The average inspection time of each case of the Diatom nitric acid digestion method was (95.30 +/- 2.78) min less than (325.33 +/- 14.18) min of plankton 16S rDNA PCR method (P < 0.05). The detection rates of two methods for field water and lung were both 100%. For liver and kidney, the detection rate of plankton 16S rDNA PCR method was both 80%, higher than 40% and 30% of diatom nitric acid digestion method (P < 0.05), respectively.@*CONCLUSION@#The laboratory testing method needs to be appropriately selected according to the specific circumstances in the forensic appraisal of drowning. Compared with diatom nitric acid digestion method, plankton 16S rDNA PCR method has practice values with such advantages as less quantity of samples, huge information and high specificity.

Change rules of quantity and species of diatoms in Hunhe River in Shenyang / 法医学杂志

OBJECTION@#To observe the change rules of quantity and species of diatoms in Hunhe River in Shenyang and to provide technology and scientific evidence for drowning identification and the location of drowning in forensic investigation.@*METHODS@#In 2011, different locations for collecting water samples were chosen in Hunhe River in Shenyang. Water samples were collected and variation of quantity and species of diatoms were observed every month. And variation of dominant species of diatoms was observed every week.@*RESULTS@#The quantity, species and dominant species of diatoms in Hunhe River in Shenyang varied with different time and locations. The quantity and species of diatoms were lowest from December to February and gradually increased, reaching peak in May and second peak in October, and then gradually decreased. The dominant species of diatoms varied significantly adjacent two weeks at same location from April to November, but had little changes at different locations in same week from July to August.@*CONCLUSION@#The change rules of quantity and species of diatoms are complicated and affected by various factors such as environment and hydrology. The change rules of species and quantity of diatoms should be considered in forensic investigation of drowning identification and the location of drowning.

Progress in application of microbeam X-ray fluorescence spectroscopy in forensic science / 法医学杂志

Microbeam X-ray fluorescence (micro-XRF) spectrometry has been raised as an analytical technique of microbeam during the recent years. With its advantages of high sensitivity, small sample requirement, high testing accuracy and non-destruction, the technique is widely utilized in forensic science. This review bases on recent researches at home and abroad, describes its applications including identification of gunshot residue, visualization of fingerprints, discrimination of drug source, production process, and other material evidences of analysis in crime scene. Thanks to the advances in technology, intelligent and portable micro-XRF equipment has appeared to be applied. It is believed that it may be more popular and frequent in administration of forensic science in the near future.

Diatoms: Role in drowning.

Drowning is a form of asphyxia due to aspiration of fluid into air passages caused by submersion in water or other fluid and is mostly accidental. The main question in case of a body recovered from water is whether the individual was alive at the time he entered water. If a body is found in water is does not necessarily mean, that this person has drowned.Deaths by drowning are difficult to determine and are often diagnosed by eliminating all other potential causes of death. Diatoms found inside the body of a drowned victim may serve as corroborative evidence in the diagnosis of cause of death. It can be ascertained whether the drowning is ante-mortem or post-mortem. The diatom test stands as the only direct screening test for drowning. The present case is one such example wherein the presence of diatoms has helped in arriving at a decision of ante mortem drowning, thereby emphasising its significance.

Diatom test in lung tissue of corpses in water and causes of death / 法医学杂志

OBJECTIVE@#To explore potential application of diatom test of lung tissue in investigation of cause of death in victim found in the water.@*METHODS@#Four hundred and seven cases were collected and analyzed for cause of death and the nature of case. Diatom test was performed in tissues and the amount was quantified. Forty-five rabbits died in the water (antemortem, postmortem and different seasons drowning) were randomly divided into 9 groups and the diatom content in lung tissue were tested with the method of nitric acid.@*RESULTS@#In 407 drowning cases, 372 cases showed a positive result of diatom test. In positive cases, the amount of accidents or suicide were 35 and homicide were 21. Thirty-five cases showed negative result of diatom test and majority were homicide in which bodies were thrown into the water after killing. Some drowning cases were in special circumstances. Animal experiments confirmed that a large amount of diatoms in lung tissue were detected in drowning victim and showed the same type in water. The amount of diatom in lung tissue was usually lower in the summer and winter comparing with spring and autumn.@*CONCLUSION@#The diatom test could be considered as an auxiliary evidence in determination of cause of death in body found in the water. The test results were related with manner of death. The nature of case should be synthetically determined based on autopsy, criminal scene and details of the cases, etc.

Determination of acid-resistant silicic particles in lung by micro-XRF and its application in diagnosis of drowning / 法医学杂志

OBJECTIVE@#Using microbeam X-ray fluorescence (Micro-XRF) analyzer for determination of acid-resistant silicic particles in lung, and to explore its potential application in diagnosis of drowning.@*METHODS@#Thirty two white rabbits were divided randomly into drowning group (n=12), post-mortem immersion group (n=10) and control group (n=10). Lungs and water sample were collected for determination of area concentration of acid-resistant silicic particles using Micro-XRF method.@*RESULTS@#The area concentration of acid-resistant silicic particles for the drowning water sample was 4.4 mm2/mL. For the lungs of drowning group, the post-mortem immersion group and the control group, the determined average values were (25.30 +/- 10.95) mm2/g, (1.68 +/- 0.63) mm2/g and (1.65 +/- 0.85) mm2/g, respectively, with a statistically significant difference between the drowning group and the other two groups.@*CONCLUSION@#The area concentration of acid-resistant silicic particles in lungs may be used as an indicator of drowning. The method is highly sensitive and rapid. It provides a potential application in drowning diagnosis.

Advances in research of diagnosis of drowning in forensic medicine / 法医学杂志

The current macroscopical and microscopical signs in the fresh drowning victim are non-specific. Moreover, putrefaction will vanish these autopsy findings quite rapidly. The study of additional detecting methods to diagnose drowning has become one of the hottest topics in forensic medicine. This article reviewed several methods commonly used in the determination of drowning in forensic practice. The advantages and disadvantages of various methods were also discussed.