Evaluation of a Novel 32 X-STR Loci Multiplex System in the Forensic Application

The AGCU X Plus STR system is a newly developed multiplex PCR kit that detects 32 X-chromosomal STR loci simultaneously.These are DXS6807,DXS9895,linkage group 1(DXS10148,DXS10135,DXS8378),DXS9902,DXS6795,DXS6810,DXS10159,DXS10162,DXS10164,DXS7132,linkage group 2(DXS10079,DXS10074,DXS10075),DXS981,DXS6800,DXS6803,DXS6809,DXS6789,DXS7424,DXS101,DXS7133,GATA172D05,GATA165B12,linkage group 3(DXS10103,HPRTB,DXS10101),GATA31E08 and linkage group 4(DXS8377,DXS10134,DXS7423).A major advantage of this kit is that it takes into account linkage between loci,in addition to detecting more X-STR loci.In order to evaluate the forensic application of 32 X-STR fl uorescence amplifi cation system,PCR settings,sensitivity,species specifi city,stability,DNA mixtures,concordance,stutter,sizing precision,and population genetics investigation were evaluated according to the Scientific Working Group on DNA Analysis Methods(SWGDAM)developmental validation guidelines.The study showed that the genotyping results of each locus were signifi cantly accurate when the DNA template was at least 62.5 pg.Complete profi les were obtained for the 1∶1 and 1∶3 combinations.A total of 209 unrelated individuals from Southern Chinese Han community,consisting of 84 females and 125 males,were selected for population studies,and 285 allele profi les were detected from 32 X-STR loci.The polymorphism information content(PIC)ranged from 0.2721 in DXS6800,to 0.9105 in DXS10135,with an average of 0.6798.DXS10135(PIC=0.9105)was the most polymorphic locus,with discrimination power(DP)of 0.9164 and 0.9871 for the male and female.The cumulative PD_(F),PD_(M),MEC_(trio) and MEC_(duo) valu es were all greater than 0.999999999.There were 78 different DXS10103-HPRTB-DXS10101 haplotypes among the 125 males,and the haplotype diversity was 0.9810.There was no signifi cant difference in the cumulative PD_(F),PD_(M),MEC_(trio) and MEC_(duo) values whether considering linkage or not.In summary,the new X-STR multiplex typing system is effective and reliable,which can be useful in human genetic analysis and kinship testing as a potent complement to autosomal STR typing.

The role of snapin in regulation of brain homeostasis

Brain homeostasis refe rs to the normal working state of the brain in a certain period,which is impo rtant for overall health and normal life activities.Currently,there is a lack of effective treatment methods for the adverse consequences caused by brain homeostasis imbalance.Snapin is a protein that assists in the formation of neuronal synapses and plays a crucial role in the normal growth and development of synapses.Recently,many researchers have reported the association between snapin and neurologic and psychiatric disorders,demonstrating that snapin can improve brain homeostasis.Clinical manifestations of brain disease often involve imbalances in brain homeostasis and may lead to neurological and behavioral sequelae.This article aims to explo re the role of snapin in restoring brain homeostasis after injury or diseases,highlighting its significance in maintaining brain homeostasis and treating brain diseases.Additionally,it comprehensively discusses the implications of snapin in other extracerebral diseases such as diabetes and viral infections,with the objective of determining the clinical potential of snapin in maintaining brain homeostasis.

Perception,use of social media,and its impact on the mental health of Indian adolescents:A qualitative study

BACKGROUND Mental illness is a health challenge faced by adolescents that has grown worse after the Coronavirus disease 2019 pandemic.Research on social media and young people’s mental health has recently increased,and numerous studies have examined whether frequent use of social media is linked to issues such as anxiety,stress,depression,eating disorders,insomnia,frustration,feeling alone,and externalizing problems among adolescents.This influence of social media on adolescents’lives is clear,with many platforms like Facebook,Instagram,and YouTube playing an important role in daily interactions and self-expression.Even though social media offers numerous benefits,such as connectivity and information sharing,excessive usage can have detrimental effects on mental health,particularly among adolescents.AIM To study the impact of social media on the mental wellbeing of adolescents,and the associated potential dangers in India.METHODS A total of 204 adolescents aged 14 years to 23 years were included in the study.This study explored the intricate relationship between social media usage and adolescent mental health in India.The study employs a cross-sectional survey design to capture a snapshot of adolescent mental health and social media usage patterns.Data collection involved administering structured questionnaires and the analysis utilized quantitative methods,including descriptive statistics.RESULTS Excessive use of social media is correlated with increased stress,anxiety,and depression.Adolescents engage in compulsive behaviors such as scrolling in the middle of the night,which negatively impacts their mental and physical health,and leads to significant sleep disruption.Findings from the study aim to provide insights into the current state of adolescent mental health and inform strategies to promote positive wellbeing in the Indian population.CONCLUSION The study underscores the need for further research to better understand the complex interplay between social media and adolescent mental health,and need for effective strategies to combat online harassment.

Messenger RNA Profiling for Forensic Body Fluid Identification： Research and Applications

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts.However,the conventional body fluid identification methods are prone to various limitations,such as time consumption,intensive labor,nonparallel manner,varying degrees of sensitivity and limited specificity.Recently,the analysis of cell-specific messenger RNA expression(mRNA profiling)has been proposed to supplant conventional methods for body fluid identification.Since 2011,the collaborative exercises have been organized by the European DNA Profiling Group(EDNAP)in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification.The major advantages of mRNA profiling,compared to the conventional methods,include higher sensitivity,greater specificity,the ability of detecting several body fluids in one multiplex reaction,and compatibility with current DNA extraction and analysis procedure.In the current review,we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.

Assessment of a Sudden Death Case due to Coronary Artery Disease Based on the PMCT and Forensic Autopsy

It is never an easy thing to diagnose heart vascular disease only depending on the unenhanced postmortem computed tomography(PMCT).This article reported a case of sudden natural death after the complaint of anterior chest pain in which coronary artery calcification(CAC) was clearly displayed using PMCT scan.The entire coronary artery system was almost reconstructed via multiplanar reformation(MPR) and volume-rendering reconstruction(VR),and the total calcium score of the coronary arteries was obtained with CaScoring automatic analysis software.The results showed that CAC was conspicuous;the total calcium score was 640.3,considerably higher than 400.The pulmonary ground-glass opacity(GGO) and small amount of fluid both in the subglottic trachea and main bronchi were also found.The imaging results confirmed those of autopsy.In addition,the results concluded that PMCT might serve as an invaluable adjunct to the classic autopsy procedure.

Analysis of γ-Hydroxybutyrate in Human Urine by LC-MSIMS Method and Its Forensic Application

GHB （γ-hydroxybutyrate） is becoming popular recreational drugs. As a result of its strong sedative and amnesiac effects, GHB has been implicated in a number of DFSA cases. The natural presence of GHB in the human body and its rapid elimination after ingestion make it difficult to detect and to evaluate its roles in suspected GHB-facilitated assaults. The paper describes an analytical method for the determination of GHB in urine using LC-MS/MS. Samples were acidified by ammonium chloride solution and extracted with ethyl acetate, and then the extracts were analysed by LC-MS/MS. The limit of detection was 0.05 p.g/mL （S/N = 3）. The intra- and inter-day precision was within 10.0% at three concentrations. The methods were found to be sensitive, accurate, rapid and suitable for the forensic toxicology to test GHB in real cases.

Mitophagy in intracerebral hemorrhage:a new target for therapeutic intervention

Intracerebral hemorrhage is a life-threatening condition with a high fatality rate and severe sequelae.However,there is currently no treatment available for intracerebral hemorrhage,unlike for other stroke subtypes.Recent studies have indicated that mitochondrial dysfunction and mitophagy likely relate to the pathophysiology of intracerebral hemorrhage.Mitophagy,or selective autophagy of mitochondria,is an essential pathway to preserve mitochondrial homeostasis by clearing up damaged mitochondria.Mitophagy markedly contributes to the reduction of secondary brain injury caused by mitochondrial dysfunction after intracerebral hemorrhage.This review provides an overview of the mitochondrial dysfunction that occurs after intracerebral hemorrhage and the underlying mechanisms regarding how mitophagy regulates it,and discusses the new direction of therapeutic strategies targeting mitophagy for intracerebral hemorrhage,aiming to determine the close connection between mitophagy and intracerebral hemorrhage and identify new therapies to modulate mitophagy after intracerebral hemorrhage.In conclusion,although only a small number of drugs modulating mitophagy in intracerebral hemorrhage have been found thus far,most of which are in the preclinical stage and require further investigation,mitophagy is still a very valid and promising therapeutic target for intracerebral hemorrhage in the long run.

Personalized medicine and opioid use disorder

Opioid use disorder(OUD)is a major public health problem affecting millions of people worldwide.Although OUD is a chronic and relapsing disorder,a variety of pharmacological and non-pharmacological interventions are available.Medication-assisted treatment of OUD generally relies on competition for opioid receptors against the addictive substance.The mechanisms of this competition are to block or inactivate the opioid receptor or activate the receptor with a substance that is intermittent or long acting.Methadone and buprenorphine are two United States Food and Drug Administration-approved medications that have long-term positive effects on the health of opioid-dependent individuals.Although clinical studies of drugs generally demonstrate efficacy in thousands of people and toxicity is excluded,it cannot be predicted whether the given drug will cause side effects in one of the patients at the treatment dose.Individual differences can be explained by many biological and environmental factors.Variations in genes encoding drug metabolism or cellular drug targets significantly explain the variability in drug response between individuals.Therefore,for the effects of candidate genes to be accepted and included in individual treatment protocols,it is important to repeat studies on individuals of different ethnic backgrounds and prove a similar effect.

Distal-scanning common path probe for optical coherence tomography

In this paper,we present a distal-scanning common path probe for optical coherence tomography(OCT)equipped with a hollow ultrasonic motor and a simple and specially designed beam-splitter.This novel probe proves to be able to effectively circumvent polarization and dispersion mismatch caused by fiber motion and is more robust to a variety of interfering factors during the imaging process,experimentally compared to a conventional noncommon path probe.Furthermore,our design counteracts the attenuation of backscattering with depth and the fall-off of the signal,resulting in a more balanced signal range and greater imaging depth.Spectral-domain OCT imaging of phantom and biological tissue is also demonstrated with a sensitivity of∼100dB and a lateral resolution of∼3μm.This low-cost probe offers simplified system configuration and excellent robustness,and is therefore particularly suitable for clinical diagnosis as one-off medical apparatus.

Robust and efficient direct multiplex amplification method for large-scale DNA detection of blood samples on FTA cards

Deoxyribonucleic acid (DNA) damage arising from radiations widely occurred along with the development of nuclear weapons and clinically wide application of computed tomography (CT) scan and nuclear medicine. All ionizing radiations (X-rays, γ-rays, alpha particles, etc.) and ultraviolet (UV) radiation lead to the DNA damage. Polymerase chain reaction (PCR) is one of the most wildly used techniques for detecting DNA damage as the amplification stops at the site of the damage. Improvements to enhance the efficiency of PCR are always required and remain a great challenge. Here we establish a multiplex PCR assay system (MPAS) that is served as a robust and efficient method for direct detection of target DNA sequences in genomic DNA. The establishment of the system is performed by adding a combination of PCR enhancers to standard PCR buffer. The performance of MPAS was demonstrated by carrying out the direct PCR amplification on 1.2 mm human blood punch using commercially available primer sets which include multiple primer pairs. The optimized PCR system resulted in high quality genotyping results without any inhibitory effect indicated and led to a full-profile success rate of 98.13%. Our studies demonstrate that the MPAS provides an efficient and robust method for obtaining sensitive, reliable and reproducible PCR results from human blood samples.

Reliability Study on Quantitative Detection of Extensor Digi- torum Brevis Strength with Needle Electromyography and Nerve Conduction

Objective To study the objectivity and reliability of needle electromyography and nerve conduction for detection of musculus extensor digitorum brevis strength, which may provide a basis for establishing a quantitative detection of muscle strength in forensic clinical study. Methods Forty-four healthy people were enrolled as the subjects, and during toe dorsiflexion, the following items including needle electromyography indexes, motor unit potential(MUP) amplitude, MUP count, recruitment reaction type, and nerve conduction detection indexes, compound muscle action potential(CMAP) amplitude, CMAP latent period and motor nerve conduction velocity(MNCV), were simultaneously detected under the cooperation and disguise condition. Results Under the cooperation condition, regardless of the same operator or different operators, there were good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, while there were normal test-retest reliabilities in MUP count and recruitment reaction type and the repeatability of the same operator was slightly better than the repeatability between different operators. Under the disguise condition, test-retest reliabilities of MUP amplitude, CMAP amplitude, CMAP latent period and MNCV were relatively high, while test-retest reliabilities of MUP count and recruitment reaction type were relatively low. Conclusion There are good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, which can be conducive to comparison between different operators and results at various times; MUP count and recruitment reaction type, which can be easily affected by subjectivity of operators and examinees, can be used to differentiate whether an examinee disguises or not. The indexes used to objectively judge muscle strength remain to be further investigated.

A high-quality reference genome of wild Cannabis sativa

Cannabis sativa is a well-known plant species that has great economic and ecological significance.An incomplete genome of cloned C.sativa was obtained by using SOAPdenovo software in 2011.To further explore the utilization of this plant resource,we generated an updated draft genome sequence for wild-type varieties of C.sativa in China using PacBio single-molecule sequencing and Hi-C technology.Our assembled genome is approximately 808 Mb,with scaffold and contig N50 sizes of 83.00 Mb and 513.57 kb,respectively.Repetitive elements account for 74.75%of the genome.A total of 38,828 protein-coding genes were annotated,98.20%of which were functionally annotated.We provide the first comprehensive de novo genome of wild-type varieties of C.sativa distributed in Tibet,China.Due to long-term growth in the wild environment,these varieties exhibit higher heterozygosity and contain more genetic information.This genetic resource is of great value for future investigations of cannabinoid metabolic pathways and will aid in promoting the commercial production of C.sativa and the effective utilization of cannabinoids.The assembled genome is also a valuable resource for intensively and effectively investigating the C.sativa genome further in the future.

Severe Hypokalemia due to Combination Use of Dexamethasone and Calcium Gluconate for Diabetics with Infection： A Case Report

The combination use of dexamethasone and calcium gluconate can be applied to hypersensitivity. Severe hypokalemia is a usual complication of dexamethasone and calcium gluconate therapy, which occurs frequently with therapeutic use. Fatal cases, accidental and intentional, occur frequently in forensic practice. The current case report presented a 43-year-old man with diabetes mellitus with infection, to whom dexamethasone and calcium gluconate were administered in the private clinic. With the development of such clinical symptoms of severe hypokalemia as quadriplegia, he was confirmed to have severe hypokalemia through a biochemical test before dying of arrhythmia. And also it presented pathophysiologic mechanism underlying severe hypokalemia as well as suggestions for clinical practice regarding combination use of dexamethasone and calcium gluconate.

Study of vehicle accident reconstruction based on the information of the tire marks

The tire mark is the important legacy information left at the accident scene. Based on the vehicle collision dynamics model, this study provided an optimized algorithm with vehicle final location and other related information for the tire marks. When the tire marks simulation results fit the real one well, the state of vehicle can be understood as the real state in the accident. Based on above, the vehicle velocity and direction are decided. According to the velocity and direction of the vehicle, the complete accident process can be simulated. With the help of the Pc-Crash software, the method has been applied in typical collision accident cases analysis. The reconstruction results will provide the scientific and numerical references for vehicle collision accidents analyzing and appraising.

Genome and transcriptome of Papaver somniferum Chinese landrace CHM indicates that massive genome expansion contributes to high benzylisoquinoline alkaloid biosynthesis

Opium poppy(Papaver somniferum)is a source of morphine,codeine,and semisynthetic derivatives,including oxycodone and naltrexone.Here,we report the de novo assembly and genomic analysis of P.somniferum traditional landrace‘Chinese Herbal Medicine’.Variations between the 2.62 Gb CHM genome and that of the previously sequenced high noscapine 1(HN1)variety were also explored.Among 79,668 protein-coding genes,we functionally annotated 88.9%,compared to 68.8%reported in the HN1 genome.Gene family and 4DTv comparative analyses with three other Papaveraceae species revealed that opium poppy underwent two whole-genome duplication(WGD)events.The first of these,in ancestral Ranunculales,expanded gene families related to characteristic secondary metabolite production and disease resistance.The more recent species-specific WGD mediated by transposable elements resulted in massive genome expansion.Genes carrying structural variations and large-effect variants associated with agronomically different phenotypes between CHM and HN1 that were identified through our transcriptomic comparison of multiple organs and developmental stages can enable the development of new varieties.These genomic and transcriptomic analyses will provide a valuable resource that informs future basic and agricultural studies of the opium poppy.

Simultaneous Determination of Heroin,Amphetamine and their Basic Impurities and Adulterants Using Microemulsion Electrokinetic Chromatography

Simultaneous separation of 17 species of heroin, amphetamine and their basic impurities and adulterants was conducted within 10 minutes by using capillary microemulsion electrokinetic chromatography. The influences of pH and 1-butanol cosurfactant on the separation were investigated, and 1-butanol was found to be a principal factor to improve separation efficiency.

Electroretinogram and Histopathologic Changes of the Retina after Methanol Intoxication

In order to study the functional and structural alterations of the retina in SD rat model after methanol intoxication,35 rats were divided randomly into five groups administrated with saline,3-day high dose,7-day high dose,3-day low dose and 7-day low dose methanol separately.The retinal function of each group was assessed by flash electroretinogram(F-ERG) 3 and 7 days after methanol poisoning.The microstructure and ultrastructure of the retina were observed at the same time.The high-dose methanol intoxication induced irreversible retinal functional and structural damages 3 days after poisoning,which included prolonged latency and reduced amplitude of the Max-reaction of F-ERG.These injuries were aggravated 7 days after poisoning.Meanwhile,the latency and amplitude of the Cone-reaction of F-ERG were also affected 3 days after poisoning,but there were no further worsening tendency 7 days after poisoning.The retinal histological analysis showed cellular edema,heteromorphy and disarrangement,tissular loosen of the inner nuclear layer and photoreceptors layer.The mitochondrial damage began at the photoreceptors layer and developed further into the inner nuclear layer.The low-dose methanol intoxication only caused transient damage of the retina.Our results showed that the function and structure of the photoreceptor and inner nuclear layer were the primary target of methanol intoxication and that the rod cells were more sensitive to methanol intoxication than the cone cells.The mitochondrial damage developed from outer layer to inner layer of the retina.

Stature Estimation Based on the Length of Tibia and Fibula Measured by Digital X-ray in Chinese Han Teenagers

Objective To derive regression formulae for stature estimation using forensic radiography from the tibia and fibula of Chinese Han teenagers in Sichuan Province. Methods To construct equations, measurements were conducted on the training sample (412 adults, 201 males and 211 females). The whole length of the fibula and four measurements of tibia were determined using CR radiography, rectified through theoretical magnification. The regression formulae were relatively constructed to the real stature measured in an erect position. Through using the testing sample (40 adults) for the regression formulae, the reliability of the regression formulae was assessed. Results The range of correlation coefficients of four measurements for tibia was 0.880-0.895 in the sex-unknown group, 0.869-0.893 in the male, and 0.8450.855 in the female. The five measurements were found to be better correlated with stature in the male than in the female. Conclusion The digital X-ray of the tibia and fibula for stature estimation is proved to be effective in forensic individual identification; therefore, these equations can be of great assistance to the stature estimation of the contemporary Chinese Han teenagers.

The Application of X-STR： Two Case Reports

The usefulness of the X-chromosomal STRs （short tandem repeats） for forensic purposes seems to be restricted, but may be valuable in some paternity cases. Due to the particular mode of inheritance, X-chromosomal X-STR has the advantage in female traces identification against male contamination and in complex kinship cases, such as deficiency cases （mother-son or father-daughter）, grandparent-grandchild comparisons, half-sisters testing, etc. In this study, 4 blood samples from two paternity cases without father were examined with TYPER19 Amplification kit first, from the result, it is hard to judge the relationship between the mother-sons, then TYPERX19 kit was conducted to obtain the full profiles of the two pairs of the mother-son, from the genetypes of the samples, it can be inferred that neither of the two pairs of mother-son were in line with the kinship, which provides important evidence for the two paternity cases. All together, the X-chromosome marks included in the TYPERX19 kit can offer the possibility to solve complex kinship cases where autosomal STR markers cannot provide the information needed.

Long-Term Trend of Bone Development in the Contemporary Teenagers of Chinese Han Nationality

Objective To further improve the accuracy of bone age identification using the time of secondary ossification center appearance and epiphyseal fusion of 7 joints to estimate the age of living individuals.Methods DR films were taken from 7 parts including sternal end of clavical and the left side of shoulder,elbow,carpal,hip,knee and ankle joints of 1 709 individuals who came from eastern China,central China and southern China,whose ages were between 11.0 and 20.0 years.From those 7 joints 24 osteal loci were selected as bone age indexes,which could better reflect age growth of teenagers.The characteristics of secondary ossification center appearance and epiphyseal fusion were observed,and the mean and age range of secondary ossification center appearance and epiphyseal fusion were calculated.Results The fusion time of the 24 epiphyses were advanced at different degrees,the most obvious epiphyses the sternal end of clavicle,scapular acromial end,distal end of the radius,distal end of the ulna,iliac crest,ischial tuberosity,the upper and lower end of tibia and fibula.The appearance time of sternal end of clavicle,scapular acromial end,iliac crest and ischial tuberosity epiphyses were all found to be after the age of 12,and the female's age,approximately 1 year ahead of schedule in comparison with the male's.Conclusion The relevant forensic information and data for bone age identification should be updated every 10-15 years so as to provide accurate and objective evidence for court testimony,conviction and sentencing.