New synthetic opioids are on the growing list of illicit drugs and pose a serious threat to human health. Some examples of NSOs include AP 237, piperidine, bromomorphine and a variety of 2-benzyl benzimidazole opiates. 2-benzyl benzimidazole opioids, also known as “Nitazenes”, many of which are regulated by the United Nations Commission on Narcotic Drugs, have become internationally prevalent in recent years. Metonitazene is an emerging potent synthetic opioid that first appeared in the recreational drug supply in mid-2020 and began to surge by the end of the decade, raising increasing public health concerns. In this study, we presented a method for the identification of a novel synthetic opioid metonitazene seized for the first time in China by gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography-quadrupole/electrostatic field Orbitrap-MS (UPLC-Q- Exactive Orbitrap-MS). The unknown samples were extracted by methanol, and the supernatants were analyzed by GC-MS and UPLC-Q-Exactive Orbitrap-MS respectively. Isotonitazene was analyzed as a contrast. By GC-MS detection, the main characteristic ion peaks of the mass spectral fragments of unknown component with retention time of 15.167 min were m/z 86.1 (base peak), 58.05, 121.1, 205.05, 236.05 and 248.1. UPLC-Q-Exactive Orbitrap-MS detection showed that the excimer ion peak of unknown component with retention time of 7.35 min was m/z 383.207 7. The main ions of secondary mass spectrum were m/z 310.118 2, 264.125 2, 121.064 9, 100.112 1 and 72.080 8. Due to the lack of reference substance, the target was identified as metonitazene by retrieval of SWGDRUG and Cayman database, comparison of information in reported literatures and analysis of high-resolution mass spectrometry data. The method is simple, rapid and has good application prospect. It can be used in real case detection.

Artificial intelligence (AI) is a comprehensive new region developed by computer science, neuroscience, linguistics and other disciplines. It is an intelligent solution based on big data, machine learning and other technologies, which can improve morphological recognition ability, diagnostic efficiency and quality. With the rapid development of science and technology, AI technology has made great progress and been widely used, providing new solutions for solving various practical problems. “AI+” has been widely used in all walks of life and achieved excellent results, among which, a number of excellent research results have emerged in the field of forensic medicine. In recent years, forensic scholars at home and abroad have studied the application of AI technology in many aspects, such as postmortem interval estimation, diatom test, age and sex estimation, DNA map analysis, poison test and injury mechanism determination. The achievements have greatly promoted the progress of forensic science and demonstrated the advantages of applying AI technology to solve traditional forensic problems. This paper summarizes the literatures in recent three years, hoping to provide new ideas for the research of forensic pathology, clinic, anthropology and toxicology.

Etomidate is a hypnotic agent used for induction of anesthesia, characterized by rapid onset, short duration, and hemodynamic stability. It is widely used for anesthesia induction in elderly patients and those with cardiovascular disease and critical illness. The “2021 China Drug Situation Report” issued by the National Narcotics Control Commission pointed out that there is a situation of abuse of “smoke powder” in some areas, and its main component, etomidate, is identified as a drug substitute. However, there have been no reports of etomidate-related deaths in China. This article reports a case of a minor who died from respiratory failure due to exacerbation of bronchopneumonia after inhaling etomidate, for reference by peers. In forensic practice, attention should be paid to the adverse reactions of etomidate and the abuse of etomidate.

In recent years, with the continuous improvement of public security conditions, the incidence of homicides has significantly decreased, but the occurrence of unnatural deaths remains high and is showing a growing trend. Death from hanging is one of the common causes of unnatural death cases, the key point is identifying suicide or homicide and the difficulty is to judge the manner of death. Homicide from hanging is uncommon. This article introduces the situation of body damage and scene investigation about a typical case of homicide from hanging yet being pretended of suicidal, analyzes the characteristics of suicide and homicide from hanging, and discusses the key points for handling similar cases to provide reference for judgment of death cause, determination of manner of death, and identification of investigation direction in handling related cases.

This article introduces a case of asphyxial death caused by using plastic ties to tie (strangle) the neck. The deceased was a teenage female who lost contact with her family while climbing the mountain alone. The body was found on the mountain the next day. The deceased’s hands, feet, and neck were tied with plastic ties. After removing the ties, a 0.8cm wide circular closed cable groove was exposed on the neck. Mild congestion on the face of the deceased, patchy bleeding on the bulbar conjunctiva, more mushroom-like foam in the mouth, nose and trachea, no significant bleeding under the neck sulcus and muscles; Forensic histopathology revealed a decrease in the stratum corneum of the cervical sulcus skin, narrowing of cell spacing, elongation of cells and nuclei, pulmonary congestion and bleeding, focal interstitial bleeding in the heart, wave like changes in some myocardial fibers, edema of brain cells, and hypoxic changes in neurons; No toxic substances such as ethanol, barbital, phenobarbital, scobarbital, and tetramine were detected in the physical and chemical tests. Based on the above analysis, the deceased was found to have died from asphyxia due to external compression of the neck caused by plasticties. According to the analysis of signs such as more mushroom-like foam in the mouth, nose and trachea of the deceased, congestion in the face and mild bleeding, the neck bandage failed to completely close bilateral carotid arteries, veins and trachea, indicating that the death process of the deceased lasted for a long time. In addition, there is a difference of several hours between the estimated time of death using indicators such as the degree of digestion of gastric contents in this case and the time of crime confessed by the suspect. Through reviewing relevant literature, it was found that there is a significant error in using the degree of digestion of gastric contents to estimate the time of death in individual cases, which is related to factors such as personal constitution, on-site environment, and dietary conditions, It is suggested that forensic colleagues should not overly rely on a single indicator when inferring the time of death using early corpse phenomena. Instead, they should analyze factors such as the case, on-site environment, dietary conditions, and personal physique as comprehensively as possible.

Palm print is one of the most important parts of dermatoglyphics and forensic science. Palm print is genetic related and also is individual unique during one’s whole life. It has played and still playing a very important role in the individual identification. In this paper 720 pairs of inked palm print samples were collected and studied. Twenty two types of papillary patterns were found and their frequency of occurrence in each areas of palm was calculated. The distribution characters of triangles in tri-radiate area and the other accompanying patterns, such as stairs-like ridges and small upside-down loop in thenar area, were analyzed. The combined patterns formed in tri-radiate, thenar and hypothenar area were summarized. The characters of palm print patterns in the three areas were also discussed according to gender and hands. The result showed that three to seven triangles could be formed in tri-radiate area of palm and the number of triangles was related closely to the type and quantity of papillary patterns appearing in this area. Ninety-six point eight seven percent of the tri-radiate prints had one to three papillary patterns, in which the upside-down loop beneath ring finger and little finger has the highest overall frequency. Eight types of single papillary patterns and ten types of double combined papillary patterns have been found in the tri-radiate area, while three types of triple combined papillary patterns have been found in this area. The total occurrence frequency of the papillary patterns in the tri-radiate area was that of male higher than female and in right hand higher than in left hand. Nine point one zero percent of the thenar palm prints had papillary patterns, in which the inner sloping loop showed the highest frequency of occurrence. Stairs-like ridges were found above all of the inner sloping loops and outer horizontal loops, while only few percentage of small upside-down loops appearing above the stairs-like ridges in thenar area. The total occurrence frequency of the papillary patterns in the thenar area was that of male higher than female and in left hand higher than in right hand. Fourteen point one seven percent of the hypothenar palm prints had papillary patterns, in which the outer sloping loop showed the highest frequency of occurrence. Eight types of single papillary patterns and three types of double combined papillary patterns have been found in the hypothenar area. The occurrence frequency of the papillary patterns in the hypothenar area was that of female higher than male and in left hand higher than in right hand. Due to the limited scope of sample collection, the statistical analysis of parameters such as the types and quantities of papillary patterns appearing in various areas of palm in the paper may not be comprehensive enough. As a result, there might be small or large deviations in the calculation of various frequency values, which will be supplemented and corrected in future research.

Choking is asphyxial death caused by a foreign body blocking the respiratory tract from the inside and obstructing the exchange of gas. In forensic pathology identification, choking is rare, and its identification is mainly based on the discovery of foreign bodies in the throat, trachea, bronchus and other parts of the deceased during autopsy, which can be food, such as lumps, or non-food. There have been previous reports of cases of choking caused by mistakenly swallowing balloons. The similarity with this case is that foreign bodies may have one-way valve effect, but the difference is that the parts of the foreign bodies obstructing the airway are different. The foreign body in this case was dried glutinous rice cake, which was mainly dry and hard, and the total amount of foreign bodies in the airway was large, which completely blocked the left and right main bronchus and caused the asphyxial death. The identification opinion of choking death must go through a systematic autopsy, exclude other causes of death, and make a comprehensive analysis and judgment in combination with the case, so as to diagnose the cause of death. The vast majority of choking deaths are accidental. This paper systematically summarized a case of accidental choking, sorted out the pathological features and points of identification with other causes of death, hoping to provide valuable reference for forensic colleagues.

In the process of investigating telecommunication network fraud cases, especially in cases such as click farming, investment and financial management fraud and naked chat, APP and URL forensics analysis are the focus of network-side investigation. Because of the need to realize functions such as chatting, picture uploading and voice calling in the APP involved, the APP developed based on IM framework has become the mainstream, among which NetEase Yunxin IM is the most common third-party IM framework in the current fraud cases. However, as criminals continue to hide their means of committing crimes, for example, encrypting APPs or encrypting chat content end-to-end, direct analysis cannot obtain the key value of IM interface, or only the encrypted garbled code can be obtained, and chat content cannot be viewed. Based on this kind of cases, this paper introduces the principle of IM framework, the encryption technology and decryption method of APPs and chat content. Through in-depth reverse analysis and encryption algorithm analysis of this kind of APPs, the efficiency of clue mining and investigation and evidence collection of single fraud cases can be fully improved, which provides strong support for the detection of related cases.

DNA test of aged skull is always a difficult problem in the field of forensic science, as the skull contains little DNA, and the DNA degrades badly. Here, an improved method is introduced for DNA test of aged skulls. Taking the skulls in recent cases for example, the optimization was carried out from the selection of materials, concentration and recovery of demineralized solution, high volume extraction and purification of DNA, etc. Finally, the results of autosomal and Y chromosome STR polymorphism were obtained successfully. The results showed that petrosal part of temporal bone could be the preferred extraction site for the test of aged skulls, with a higher success rate compared with other parts of the skull; in addition, the use of Amicon Ultra-15 10K centrifugal ultrafiltration tubes could remove a large number of small molecules, such as water and ions in the decalcification solution, so as to retain and recycle the large molecules of DNA, which effectively reduced the amount of DNA loss; furthermore, the quality of DNA was poor in aged skulls, so increasing the amount of bone powder used and increasing the extraction and purification system could also significantly improve the amount of DNA recovered. This method improves the quantity and quality of DNA recovered from aged skull, and can provide reference for the follow-up DNA test of similar aged bones and teeth.

Water-soluble poisons refer to poisons with high solubility in water, characterized by rapid absorption and high toxicity. They are easily distributed in various tissues and organs of the body through blood circulation, causing serious harm to human body. This article selected four common water-soluble poisons, including nitrite, cyanide, sodium fluoroacetate, and borax. It mainly reviewed the physicochemical properties, toxicological mechanisms, poisoning symptoms, and metabolic processes in the body of the four toxins, as well as the current developments and future trends of their detection methods form home and abroad. This article mainly aimed at providing references in forensic toxicology analysis of water-soluble poisons and investigating the causes of such suspected poisoning cases.

Objective and accurate forensic genetic analysis results are the basis for making accurate identification opinions. With the increasing sensitivity of testing equipment and amplification kits, the pressure to prevent laboratory pollution is increasing. Among them, the contamination of PCR amplification products is the most difficult type of pollution to prevent. This research introduces an anti-pollution amplification kit, which contains 16 short tandem repeats (STR) locus, one sex identification locus (Amel) and one Y chromosome insertion deletion locus (Indel). Anti-pollution amplification kit can obtain DNA fragment containing uracil base through STR multiplex amplification, and this type of DNA fragment can be efficiently hydrolyzed by uracil DNA glycosylase (UDG enzyme) at 50℃. An additional 50℃ incubation before each new round of PCR amplification can completely eliminate the pollution threat of previous amplification products to the results. After testing, the anti-pollution amplification kit has excellent anti-pollution ability, and uracil base does not change the DNA typing results. Its detection sensitivity does not decrease; the stability of DNA product fragments does not decrease; and subsequent electrophoresis analysis is not affected. This reagent kit can effectively eliminate contamination of amplification products.

An analytical method was established for determination of 5 common veterinary anesthetics (tiletamine, xylazole, ketamine, xylazine and zolazepam) in whole blood by using programmed temperature vaporizer (PTV) system combined GC-MS/MS. Those samples were dealt with salting-out-assisted liquid-liquid extraction (SALLE), introducing NaCl as salting-out regent and acetonitrile as extraction solvent. After that, the extracting solution was purified by QuEChERS nano column. Most acetonitrile in the solution were removed by PTV system before analyzing in GC-MS/MS for protection of gas chromatographic column. The multiple-reaction-monitoring (MRM) positive ion mode was selected for detection. Three pairs of characteristic ions were taken for qualitative analysis and external standard method was accepted for quantitative analysis. The method was verified well. The results showed that the LODs were 10-20 ng/mL and LOQs were 20-40 ng/mL. There were good linear relationships for all the analytes with correlation coefficients (r) not less than 0.998. At three spiked levels of 40, 400, and 2 000 ng/mL, the recoveries for 5 veterinary anesthetics in whole blood ranged from 71.61% to 119.10%. The intra-day precision and inter-day precision of the method evaluated at the concentration of three levels ranged from 0.34% to 14.81%, 4.03% to 14.87%, respectively. The developed method with broad research potential benefited its simple pretreatment, fine stability and highly sensitivity, which was applied in an actual case.

Clinical cases involving antibiotic poisoning and death are mainly due to antibiotic overdose or patients with a particularly sensitive constitution. In the paper, taking an illegal practice of medicine for example, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was carried out to detect lincomycin, ceftriaxone, and dexamethasone in suspected allergic deaths caused by antibiotics. Besides, combined with scene investigation, autopsy, pathological examination, and toxicology test, it was confirmed that the death was caused by an allergic reaction to antibioticsin. On the one hand, the research results can fill in the blank of simultaneous determination of lincomycin, ceftriaxone and dexamethasone in human plasmas by LC-MS/MS and fill in the data of death due to antibiotic allergy, and on the other hand, the research results can provide methods and data references for the clinical and forensic identification of antibiotics allergy and death cases. Furthermore, this article has summarized the forensic indications and tests of antibiotic allergy deaths, which can provide reference for the investigation of such cases and forensic toxicology.

Forensic medicine is an applied discipline that uses medical and other related knowledge to provide scientific basis and evidence for criminal investigations, civil disputes, medical disputes, and other related fields involving law, to achieve justice and maintain social harmony and stability. Time-related estimations (TRE), such as estimations of time since death, wound age, and body fluid stain age, are important components of forensic identification, which can provide clues for case investigation, delineation of investigation scope, determination of crime time, and screening of alibis. Epigenetic has received much research in many fields and is highly valuable in forensic medicine because of its special mode of inheritance that is not based on changes of DNA sequences. Non-coding RNA (ncRNA) is currently regarded as an emerging epigenetic marker in a variety of fields, and the function and mechanism of ncRNAs in the physiological and pathological processes of different systems, organs, tissues, and cells revealed. Generally, ncRNAs may be divided into two groups based on their biological roles: housekeeping ncRNAs, which are critical for sustaining fundamental cellular activities, and regulatory ncRNAs, which act as regulators in cells. With the development of molecular biology, molecular genetics and bioinformatics, non-coding RNAs such as microRNA (miRNA) and circular RNA (circRNA) have shown great potentials for forensic identification, and they provide new methods for solving forensic problems of TRE. In this review, we summarized the common detection methods of ncRNAs in forensic medicine, and described the research progress and application of ncRNA such as miRNA and circRNA in estimation of time since death, wound age, and body fluid stain age, which show close links among them and forensic medicine. Additionally, we disscussed the research values and application prospects of non-coding RNA in the TRE.

With the development of technology, more and more cases of chimerism have been found in paternity tests. At present, blood sample is the most commonly used for paternity testing, and a few institutions take oral swabs as samples. After ruling out the possibility of iatrogenic artificial chimerism and sample contamination, the presence of multiple alleles in different loci usually indicates congenital chimerism. However, if the chimera blood contains only one type of cell population, the autosomal STR typing of the blood sample will not be abnormal, and it is easy to draw a false identification conclusion that excludes relatives. This paper reports a very rare case of chimera. In this case, gene chimerism was detected only in seminal stain, and no chimerism was found in blood samples, hair and saliva spots. When blood was used for paternity test, the tested father was excluded as the biological father of the two children. However, when sperm plaques were used as samples, it was supported that the tested father was the biological father of the two children. The analysis suggests that the father is a chimera formed by the parthenogenetic division of the mother’s germ cells. Cases of chimerism, which occurs after the fusion of two fertilized eggs, have been reported only in germ cells and not in blood cells. For forensic identification, it is necessary to improve the understanding of chimerism. When the phenomenon of chimerism is suspected, the samples from different parts should be extracted and compared as far as possible.

Body fluid stains are common biological materials at crime scenes. The accurate determination of their tissue sources can help with crime scene reconstruction, case nature determination and trial. The analysis of cell-specific mRNA expression has been proposed as promising method for the identification of body fluids. Conventional strategy of mRNA profiling requires reverse transcription, PCR amplification, and electrophoresis. The one-step RT-PCR detection technology can complete reverse transcription and PCR of mRNA in one reaction, which can reduce experimental time and simplify experimental operations. In this study, we subjected the one-step multiplex reverse transcription PCR strategy to mRNA profiling with the inclusion of 10 tissue specific biomarkers in the F13plex system targeting peripheral blood (HBA, HBB), menstrual blood (MMP7, MMP10), vaginal secretion (HBD1, CYP2B7P), saliva (STATH, HTN3) and semen (PRM2, SEMG1), and 3 housekeeping genes (ACTB, GAPDH and RPL19). We verified the system’s specificity, sensitivity, and ability to detect mixed and aged samples. In terms of specificity, most of the selected genes had good specificity, but there were some cross-reactions that were hard to avoid. In order to ensure the accuracy of identification, we determined that the target body fluid was contained only when the three housekeeping genes and two specific mRNA markers were simultaneously detected. In terms of sensitivity, we found that different types of samples had different sensitivities. For example, when using 10 ng RNA for vaginal secretions and menstrual blood samples, some specific target genes were not detected and could not be correctly determined; for blood samples, even when 0.01 ng RNA was used, the RFU value of the target gene is still above 10 000. However, there are only a small amount of test materials in actual cases; it is difficult to quantify the extracted RNA. For five kinds of body fluids, 2 μL of RNA extracted from a 1 mm2 sample could all detect housekeeping genes and corresponding target genes, and the correct body fluid could be determined (Except 1 vaginal secretion sample). The target genes of the mixed components were detected in all 16 mixed samples, and correct mixed component determinations could be made, proving the system’s good ability to identify mixed samples. Almost all housekeeping genes could be detected in 14 aged samples, but only 7 were correctly identified. Because no specific target genes were detected in the remaining samples, correct determinations could not be made. Consequently, the system needs to be further optimized. For HBD1 and MMP7 with poor specificity, other vaginal secretion and menstrual blood-specific genes will be screened and verified for replacement. The instability of mRNA results in relatively poor test results for aged samples. In practical applications, other genetic markers with better stability should be used to determine the results. In general, lots of studies have been demonstrated the usability of mRNA profiling to the identification of forensic relevant body fluid. According to the comprehensive assessment of the one-step RT-PCR strategy in this study, the one-step profiling assays can be a reliable and economical method for the simplified, accurate, and simultaneous analysis of tissue-specific biomarkers for the discrimination of body fluid origin. It shows good application potential in forensic body fluid identification work.

In this paper, the characteristics of allele typing deletion of Y-STR loci and its correlation with AZF (azoospermia factor) deletion were discussed, which could provide reference for forensic practice. Y-STR kits (Yfiler Platinum, SureID PathFinder Plus) were used to analyze the blood samples of 23 461 male family members. A total of 14 cases with 4 or more Y-STR allele-dropout samples were found. Meanwhile, Sequence-tagged site (STS) was detected with Y chromosome microdeletion detection kit, and the deletion of AZF region of Y chromosome was evaluated according to the deletion STS. The results showed that the proportion of multiple Y-STR typing missing was 0.059 7% (14/23461), with 1 case of short arm and 13 cases of long arm, which were from different families with different types. STS deletion was detected in the AZF region in 13 long arm multiple typing deletion samples, and no abnormality was detected in 1 short arm multiple typing deletion sample. This study suggests that there is a correspondence between multiple STR allele dropouts in the long arm of Y chromosome and microdeletions in AZF region, and the biological basis of sterility exists in these typing individuals.

Recently, there have been instances of corporate financial personnel being defrauded in QQ group chats. The primary method employed by the criminals involves illegally obtaining QQ login permissions to gain control over user group operations. They then add the compromised account to a pre-set “work” group and wait for the account owner to log in. Upon logging in, the victim would find familiar contacts, such as his boss, in the pre-set group and receive instructions to transfer funds, ultimately resulting in fraud. Through our investigation and analysis of a real case, we discovered that these types of scams are spear-phishing attacks executed through the leakage of QQ account ClientKey information, supported by a corresponding QQ gray industry. This paper, using the evolution of QQ gray market as a backdrop, provides a detailed analysis of the complete technical architecture of this gray market. It presents methods for inspecting trojans that steal account information and mirrored servers, including ways to bypass the challenges posed by disguised source code settings in mirrored server images, and summarizes key evidence points. Lastly, through local co-debugging, we verified the coupling of the account-stealing trojans and mirrored servers, while also highlighting the security risks inherent in the current QQ fast login feature to a certain extent.

The key and difficult points of forensic analysis of high-falling cases are to infer and reconstruct human falling trajectory and analyze the formation mechanism of injury. These results help to provide technical support for the comprehensive judgment of the nature of human fall, and help to provide intuitive and effective explanations for non-professionals such as family members of the deceased. Numerical simulation technology is a series of new interdisciplinary technology formed on the basis of the continuous development of computer simulation. In recent years, thanks to the development of numerical calculation methods and the improvement of computer performance, the technology has made some progress in the research field of damage biomechanics. Numerical simulation technology has advantages of objectification, visualization, parameterization and repeatability of simulation results. This technique opens up a new way to analyze and study the fall process and fall damage in high-falling cases. Based on this kind of technology, the simulation analysis of human falling trajectory and falling damage is conducive to more scientific, objective and understandable analysis conclusions, so as to provide important technical support for judging the nature of high-falling cases, answering questions of family members and resolving social conflicts. There are two kinds of numerical simulation techniques suitable for high falls. One is to analyze and reconstruct the falling process based on the multi-rigid body dynamics method. Another is to analyze the formation mechanism of falling damage based on the finite element method. This paper reviewed the application of numerical simulation technology in high falling cases at home and abroad in recent years, and showed the application potential of this technology in the forensic analysis of high fall cases.

Hydatidiform mole (HM) is a kind of gestational trophoblastic disease caused by abnormal fertilization. The presence of abundant paternal genetic information plays a key role in the pathogenesis of hydatidiform mole. Paternity testing involving hydatidiform mole tissue is relatively rare in forensic practice. In the case discussed, the aborted tissue originating from a female victim under 14 at the time of her pregnancy was diagnosed with hydatidiform mole. It was morphologically consistent with hydatidiform mole, and showed typical genotyping features of a complete hydatidiform mole (CHM) fertilized by single sperm, i.e. homozygous in all 23 loci. The alleged suspect was not actually the biological father of the aborted embryonic tissue. The author consulted a large number of literatures to analyze the experimental results. Unexplained paternity incompatibility suggests a different biological father, who was finally found through comparison and retrieval in the DNA database. Further STR testing was performed using VeriFiler^TM Plus kit, SureID^® X37 kit and Yfiler^TM. Platinum Casework kit. One allele was detected in 23 autosome and 35 X chromosome STR loci from embryonic tissue, which was found in the genotype of the focused suspect. The STR typing technology could determine the type of hydatidiform mole and provide clues and evidence for the identification. This paper may provide enlightenment for the handling of similar cases.

Tetracaine is a common aromatic ester local anesthetic. Since the early 1930s, tetracaine has been wildly used in clinical practices, mostly for surface anesthesia, conduction anesthesia, spinal anesthesia, epidural anesthesia, but generally not for infiltration anesthesia. Yet, tetracaine can trigger off a lot of toxic reactions to the central nervous system and cardiovascular system in the course of medication. In addition, newborns with immature metabolic systems, the elderly, people with renal dysfunction and cardiac diseases and hepatic dysfunction, and pregnant women are high risk groups for local anesthetic systemic toxicity. In the last few years, with the rise of medical plastics and beauty industry, tetracaine has been gradually applied to cosmetic surgery, eyebrow tattooing, tattoos, and other fields. Nevertheless, the medical malpractice or cases of poisoning, shock and even death caused by improper operation of drugs into blood vessels, excessive dosage, and allergic reactions to drugs happen from time to time. Consequently, that not only brings a great threat to public health, but also gives rise to a large amount of hidden troubles for social stability. In current judicial practice, the cause of poisoning or death is usually inferred from the amount of drugs in the whole blood/heart blood sample or other biological tissues of the poisoned or deceased. Since tetracaine is rapidly hydrolyzed to metabolites (4-(butylamino) benzoic acid) by pseudocholine esterase in human blood, detection of tetracaine in human tissues or blood samples has been considered difficultly in practical cases. According to the above reasons, we review the research progress of tetracaine about its pharmacology, toxicological effects, metabolism in vivo, forensic toxicokinetics, metabolomics, and analytical technique in this article, which will be of reference value for forensic toxicological researchers. In the meantime, it is necessary to discuss the direction of in-depth study of tetracaine so as to provide reference for forensic toxicologist.