Effects of ketamine on mouse hippocampal inflammatory cytokines IL-6,IL-1β and TNF-α levels in acute and chronic administration models

OBJECTIVE Ketamine is an injectable anesthetic and recreational drug of abuse commonly used worldwide. Many experimental studies have shown that ketamine can impair cognitive function and induce psychotic states. Neuroinflammation has been suggested to play an important role in neurodegeneration. Meanwhile,ketamine has been showed to modulate the levels of inflammatory cytokines.Therefore,we sought to investigate whether the effects of ketamine on the central nervous system is associated with the inflammatory cytokines. METHODS We established acute(single or multiple intraperitoneal injection) and chronic(six months daily intraperitoneal injection) ketamine administration models in C57BL/6 mice,evaluated the spatial recognition memory and emotional response by applying the Y maze test and open field test. We analyzed the changes of inflammatory cytokines interleukin-6(IL-6),interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) levels in mouse hippocampus,employing Western blot,quantitative reverse transcriptase-polymerase chain reaction(qR T-PCR) and immunohistochemistry. RESULTS Ketamine induced spatial recognition memory deficit,reduced anxiety-like behaviors in mice after chronic administration,and it was dose-dependent. Moreover,we found that ketamine could increase the levels of mouse hippocampal inflammatory cytokines IL-6 and IL-1β after single,multiple and long-term administration in a dose-dependent manner. However,the level of TNF-α expressed differently in mouse hippocampus under different conditions. Single administration of ketamine increased the level of TNF-α,whereas multiple and long-term administration decreased it significantly. We considered that TNF-α might exist bi-directional regulatory pathway,which was associated with the dose and duration of ketamine administration. CONCLUSION Our results suggest that the alterations of inflammatory cytokines IL-6,IL-1β and TNF-α levels may be involved in the neurotoxicity of ketamine.

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.

Bacterial flora of conjunctiva after death

AIM:To evaluate the frequency of bacterial flora of conjunctiva after death(cadaver eyes) which will give information about the bacterial contamination of donor eyes,and the in-vitro sensitivity of isolated bacteria to the commonly used antibiotics in ophthalmic practice.METHODS:Conjunctival swabs were taken from the cadavers(motor vehicle accident deaths and patients who died in the hospital),within 6h after death,and sent for culture and sensitivity test.Conjunctival swabs,taken from the healthy conjunctiva of patients admitted for cataract surgery,were sent for culture and sensitivity as controls(eyes in those of living status).The bacterial isolates were tested against the commonly used antibiotics(chloramphenicol,gentamicin,ciprofloxacin) in ophthalmology practice.RESULTS:Bacteria were isolated in 41 out of 100 conjunctival swabs(41%),taken from 50 cadavers(study group).Coagulase negative staphylococcus was the most common bacteria isolated(15%),followed by pseudomonas aeruginosa(5%).Gentamicin was effective against majority of the bacterial isolates(82%).Bacteria were isolated from 7 out of 100 conjunctival swabs taken as control group(eyes in living state).Coagulase negative staphylococcus was the most common organism(5%) isolated in control group;the others were staphylococcus aureus(1%) and beta hemolyticus streptococci(1%).CONCLUSION:Bacteria were isolated from 41% of the cadaver eyes.High percentage sensitivity of the bacterial isolates to gentamicin(82%) supports the practice of thorough irrigation of the eyes with gentamicin solution before starting the procedure of enucleation followed by immersion of the enucleated eyeballs in gentamycin solution,to prevent the bacterial contamination.

Time-dependent Appearances of Myofibroblasts during the Re- pair of Contused Skeletal Muscle in Rat and Its Application for Wound Age Determination

Objective To research the relation between the time-dependent appearances of myofibroblasts during the repair of contused skeletal muscle in rat and wound age determination. Methods A total of 35 SD male rats were divided into the control and six injured groups according to wound age as follows: 12 h, 1 d, 5 d, 7 d, 10 d and 14 d after injury. The appearances of myofibroblasts were detected by HE staining, immunohistochemistry and confocal laser scanning microscopy. Masson's trichrome staining was utilized to examine collagen accumulation in the contused areas. Results Immunohistochemical staining showed that α-SMA+myofibroblasts were initially observed at 5 d post-injury. The average ratio of myofibroblasts was highest at 14 d post-injury, with all samples, ratios more than 50%. In the other five groups, the average of α-SMA positive ratios were less than 50%. The collagen stained areas in the contused zones, concomitant with myofibroblast appearance, were increasingly augmented along with advances of posttraumatic interval. Conclusion The immunohistochemical detection of myofibroblasts can be applied to wound age determination. The myofibroblasts might be involved in collagen deposition during the repair of contused skeletal muscle in rat.

Pleomorphic hepatocellular carcinoma following consumption of hypericum perforatum in alcoholic cirrhosis

Hepatocellular carcinoma (HCC) often develops in patients with underlying liver disease, yet HCC with syncytial giant cells (SGCs) is extremely rare. Herein, we report a 55-year-old man with a 6-year history of alcoholic cirrhosis who during his regular checkup presented with marked elevation of alpha-fetoprotein. Clinical examination and imaging analyses revealed a tumor-like lesion in segment 4 of the liver, which was removed by limited wedge resection. Histological analysis by hematoxylin and eosin staining indicated pleomorphic and atypical nodules, with some SGCs, embedded within the boundaries of the neoplastic lesion. The adjacent liver parenchyma showed microvesicular steatosis, pericellular fibrosis, and moderate hemosiderin accumulation (grade 2, as determined by Prussian blue iron stain) in hepatocytes and Kupffer cells but no copper accumulation (as determined by orcein stain). Immunohistochemical analysis showed hepatocyte antigen-positive staining for the neoplastic cells and SGCs. The diagnosis was made for cirrhosis-related HCC with SGCs. The previous reports of pleomorphic HCC have featured osteoclast-like (i.e., mesenchymal type) giant cells, making this case of epithelial type giant cells very rare. The patient’s 6-month history of hypericum perforatum/St John’s wort self-medication may have prompted the cirrhosis or HCC progression or the unusual SGC manifestation.

The model of pulmonary embolism caused by autologous thrombus in rabbits

Objective:To establish a model of pulmonary embolism in rabbits by using autologous thrombosis of rabbit ear vein, to study the method of establishing acute pulmonary embolism by using autologous thrombus and to explore the diagnostic value of oxygen partial pressure in acute pulmonary embolism.Methods: Twenty rabbits were randomly divided into normal group (n=5), 7 h group, 24h group, 1 week after model establishment Group. The arterial blood gas analysis was performed on the carotid arteries of rabbits at 7 h, 24 h and 1 W after modeling.Results: Normal group oxygen partial pressure (93.15 ± 2.26) mmHg, 7 h group oxygen partial pressure (81.98 ± 1.94) mmHg, 24 h group oxygen partial pressure (84.55 ± 2.18) mmHg, 1 W group oxygen partial pressure (92.66 ± 1.92) mmHg. Normal group oxygen partial pressure and 7 h group, 24 h group oxygen partial pressure, P value was less than 0.05 and less than 0.01, indicating that the difference was statistically significant. Normal group oxygen partial pressure and 1 week group oxygen partial pressure, P value greater than 0.05, indicating that the difference was not statistically significant.Conclusion: The oxygen partial pressure was reduced at 7 h after the establishment of the acute pulmonary embolism model and failed to return to normal within 24 h. After 1 week, the embolus began to dissolve, the respiratory and circulatory system was reestablished, and the oxygen partial pressure gradually Return to normal level. Indicating that there is a positive correlation between oxygen partial pressure and acute pulmonary embolism.

Immunohistochemical Study on Morphine in Human Tissues from Cases of Opiate Associated Death

The purpose of this investigation was to offer evidence that immumohistochemical staining is applicable to the determination of morphin distribution in human tissues from cases of opiate associated death. We studied 4 cases of opiate associated death and 2 cases of sudden cardiac death without a history of drug abuse as control. The samples were taken from cerebral cortex

Application and Prospect of Postmortem Imaging Technology in Forensic Cardiac Pathology:A Systemic Review

Postmortem imaging(PMI)technology known as virtual autopsy or virtopsy is regarded as a useful method of noninvasive or minimally invasive autopsy in forensic practice.Postmortem computed tomography is applicable to traumatic investigation,acute hemorrhage,pulmonary parenchyma disease,calcification(calculus,atherosclerosis),and gas accumulation.Postmortem magnetic resonance(PMMR)has been proven to have advantages in soft tissue identification.Cardiac death is one of the keys and difficult points in forensic practice.With the introduction and development of PMCT angiography and PMMR,it was proved to be a very promising tool in the investigation of cardiac death,including vascular cavities and ischemic myocardium.This article reviewed the applications of the latest PMI and its related technologies in forensic cardiac pathology,including advantages,limitations,and development prospects.

Glucagon-like peptide-1/glucose-dependent insulinotropic polypeptide dual receptor agonist DA-CH5 is superior to exendin-4 in protecting neurons in the 6-hydroxydopamine rat Parkinson model

Patients with Parkinson's disease(PD) have impaired insulin signaling in the brain. Incretin hormones, including glucagon-like peptide-1(GLP-1) and glucose-dependent insulinotropic polypeptide(GIP), can re-sensitize insulin signaling. In a recent phase II clinical trial, the first GLP-1 mimic, exendin-4, has shown reliable curative effect in patients with PD. DA-CH5 is a novel GLP-1/GIP receptor unimolecular coagonist with a novel peptide sequence added to cross the blood-brain barrier. Here we showed that both exendin-4 and DA-CH5 protected against 6-hydroxydopamine(6-OHDA) cytotoxicity, inhibited apoptosis, improved mitogenesis and induced autophagy flux in SH-SY5Y cells via activation of the insulin receptor substrate-1(IRS-1)/alpha serine/threonine-protein kinase(Akt)/c AMP response element-binding protein(CREB) pathway. We also found that DA-CH5(10 nmol/kg) daily intraperitoneal administration for 30 days post-lesion alleviated motor dysfunction in rats and prevented stereotactic unilateral administration of 6-OHDA induced dopaminergic neurons loss in the substantia nigra pars compacta. However, DA-CH5 showed curative effects in reducing the levels of α-synuclein and the levels of pro-inflammatory cytokines(tumor necrosis factor-α, interleukin-1β). It was also more effective than exendin-4 in inhibiting apoptotic process and protecting mitochondrial functions. In addition, insulin resistance was largely alleviated and the expression of autophagy-related proteins was upregulated in PD model rats after DA-CH5 treatment. These results in this study indicate DA-CH5 plays a therapeutic role in the 6-OHDAunilaterally lesioned PD rat model and is superior to GLP-1 analogue exendin-4. The study was approved by the Animal Ethics Committee of Shanxi Medical University of China.

Vitality and wound-age estimation in forensic pathology:review and future prospects

Determining the age of a wound is challenging in forensic pathology,but it can contribute to the reconstruction of crime scenes and lead to arrest of suspects.Forensic scholars have tended to focus on evaluating wound vitality and determining the time elapsed since the wound was sustained.Recent progress in forensic techniques,particularly high-throughput analyses,has enabled evaluation of materials at the cellular and molecular levels,as well as simultaneous assessment of multiple markers.This paper provides an update on wound-age estimation in forensic pathology,summarizes the recent literature,and considers useful additional informa-tion provided by each marker.Finally,the future prospects for estimating wound age in foren-sic practise are discussed with the hope of providing something useful for further study.

Use of deep learning in forensic sex estimation of virtual pelvic models from the Han population

Accurate sex estimation is crucial to determine the identity of human skeletal remains effectively.Here,we developed convolutional neural network(CNN)models for sex estimation on virtual hemi-pelvic regions,including the ventral pubis(VP),dorsal pubis(DP),greater sciatic notch(GSN),pelvic inlet(PI),ischium,and acetabulum from the Han population and compared these models with two experienced forensic anthropologists using morphological methods.A Computed Tomography(CT)dataset of 862 individuals was divided into the subgroups of training,validation,and testing,respectively.The CT-based virtual hemi-pelvises from the training and validation groups were used to calibrate sex estimation models;and then a testing dataset was used to evaluate the performance of the trained models and two human experts on the sex estimation of specific pelvic regions in terms of overall accuracy,sensitivity,specificity,F1 score,and receiver operating characteristic(ROC)curve.Except for the ischium and acetabulum,the CNN models trained with the VP,DP,GSN,and PI images achieved excellent results with all the prediction metrics over 0.9.All accuracies were superior to those of the two forensic anthropologists in the independent testing.Notably,the heatmap results confirmed that the trained CNN models were focused on traditional sexual anatomic traits for sex classification.This study demonstrates the potential of AI techniques based on the radiological dataset in sex estimation of virtual pelvic models.The excellent sex estimation performance obtained by the CNN models indicates that this method is valuable to proceed with in prospective forensic trials.

The forensic pathological analysis of sport-related sudden cardiac death in Southern China

Studies regarding sport-related sudden cardiac death(SCD)mainly focus on competitive ath-letes;similar data are rare in the general population,especially in China.We conducted a retrospective study(from September 1998 to August 2013)to investigate the aetiological distribution and epidemiological features of sport-related SCD in Southern China.Selections of cases are based on details,and two subgroups were established:one was the sport-related SCD group,and the other was the disease-free accident victims group which was matched with the sport-related SCD group in gender,age and year of death.Among the 3770 sudden-death cases,1656 cases were SCD cases.A total of 65 cases(57 males)out of 1656 SCD cases were sport-related.The age range of the 65 sport-related SCD cases was from 12 to 68 years old with a mean(35.92±14.23)years old.Only two of these cases were competitive athletes.The most common circumstances of the 65 sport-related SCD cases were heavy physical labour(46.15%)and running(30.77%).The three leading forensic diag-noses were the coronary atherosclerotic disease(CAD,28 cases),cardiomyopathy(CM,14 cases)and sudden unexplained death(7 cases).CM was the most common forensic diagno-sis in those?35 years old,while CAD was the most common one in those>35 years old.Left anterior descending in which atherosclerotic plaques was most commonly found was the principal artery branch associated with sport-related SCD.There was a statistically signifi-cant difference in the weight of hearts between the 65 sport-related SCD cases and 65 dis-eases-free accidental cases.This study highlights the need to attract public attention to sport-related SCD and to issue a prevention strategy to the public,and to make the SCD-related genetic sequencing a routine tool in both forensic pathological examination and clinic screening.

Perspectives on the Construction of Forensic Autopsy Laboratories Designed to Handle Infectious Diseases

Coronavirus disease 2019 has already infected tens of thousands of people in China.The existing autopsy laboratories are not qualified to perform autopsies on individuals confirmed or suspected to have died from an infectious disease.When dealing with corpses with uncertain health conditions or unknown contact with a pathogen,the forensic medical practitioners face severe risks of being infected.To meet the requirements of performing autopsies on such bodies,ensure the health and safety of forensic medical practitioners,and study the pathophysiological changes of infected patients,it is necessary to improve the construction requirements and regulations for forensic autopsy laboratories.

Forensic Examination of Abusive Head Trauma in Child Abuse Cases

Abusive head trauma(AHT)in child abuse cases is rarely encountered in the practice of forensic examination in China,and such cases are rarely reported.The authors reviewed a large number of relevant domestic and foreign studies to differentiate between the definitions of AHT and shaken baby syndrome;determine the relationship between the two;and discuss their epidemiology,mechanism,and symptoms.In addition,the main points in forensic examination and strategies for preventing AHT are also summarized to help forensic workers and clinicians identify and prevent such injuries.

Age-related changes of the redox state of glutathione in plasma

Glutathione （GSH） is the principal non-protein thiol responsible for maintaining intracellular redox status and protecting cells against oxidative/ nitrosative stresses. In the biological system, some GSH is bound to proteins, and others exist freely, including the reduced glutathione and oxidized glutathione （GSSG）. Generally it is in a steady redox state for GSH/GSSG couple to participate in a variety of physiological functions. Earlier studies showed that chemically induced oxidative stress in vivo in rats resulted in increased GSSG and decreased GSH/GSSG ratio. The impairment of the glutathione homeostasis represents an indicator of tissue oxidative stresss status in human subjects.

Estimation of the age of human semen stains by attenuated total reflection Fourier transform infrared spectroscopy:a preliminary study

Semen stain is one of the most important biological evidence at sexual crime scenes.Age estimation of human semen stains plays an important role in forensic work,and it is rarely studied due to lack of well-established methods.In this study,the technique called attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR)coupled with advanced chemometric methods was employed to determine the age of semen stains on three different substrates:glass slides,tissues and fabric made of regenerated cellulose fibres up to 6 d.Partial least squares regression(PLSR)was used in conjunction with spectral analysis for age estimation,and the results generated high R2 values(cross-validation:0.81,external validation:0.74)but a narrow margin of error for root mean square error(RMSE)(RMSE of cross-validation:0.77 d,RMSE of prediction:1.02 d).Additionally,our results indicated the robustness of PLSR model was not weaken by the influence of different substrates in this study.Our results indicate that ATR-FTIR,combined with chemometric methods,shows great potential as a convenient and efficient tool for age estimation of semen stains.Moreover,the method could be applied to routine forensic investigations in the future.

RNA-seq profiling reveals differentially expressed genes as potential markers for vital reaction in skin contusion: a pilot study

Detection of the vitality of wounds is essential in forensic practice.The present study used Illumina RNA-seq technology to determine gene expression profiles in contused mouse skin.In obtained high quality sequencing reads,the reads were mapped onto a reference transcriptome (Mus_musculus.GRCm38.83).The results revealed that there were 659 upregulated and 996 down-regulated differentially expressed genes (DEGs) in contused mouse skin.The DEGs were further analyzed using the Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes databases.Genes from different functional categories and signalling pathways were enriched,including the immune system process,immune response,defense response,cytokine-cytokine receptor interaction,complement and coagulation cascades and chemokine signalling pathway.Expression patterns of 11 DEGs were verified by RT-qPCR in mice skins.In addition,alterations of five DEGs were also analyzed in postmortem human wound samples.The results were in concordance with the results of RNA-seq.These findings suggest that RNA-seq is a powerful tool to reveal DEGs as potential markers for vital reaction in terms of forensic practices.

Diagnosis of coronary artery disease using targeted post-mortem computed tomography coronary angiography:a case report

Targeted post-mortem computed tomography(PMCT)combined with coronary angiography has the potential to play a significant role in the investigation of sudden cardiac death.The authors utilized a targeted PMCT coronary angiography in a case involving a 53-year-old man who died from acute myocardial ischemia and cardiac decompensation which may result from coronary artery disease(CAD).The victim collapsed suddenly at work and died soon after arrival to hospital.The body was examined using PMCT and targeted PMCT coronary angiography.The left anterior descending coronary artery exhibited 75%-100%stenosis in the middle segment;however,the distal segment could not be clearly visualized.In addition,the left circumflex and right coronary artery exhibited calcification,atherosclerosis and an area of 50% stenosis.Signs of cardiogenic pulmonary oedema were also identified.The imaging results suggested that this individual had coronary atherosclerosis and probably died from CAD.The autopsy and histological examination revealed acute myocardial ischemia and myocardial scarring,confirming the cause of death while excluding other probabilities.In summary,targeted post-mortem computed tomography angiography(PMCTA)can visualize the arteries and estimate the degree of principal pathological changes.This method is a simple,reliable and sensitive technique for identifying the presence of coronary atherosclerosis.It is a valuable post-mortem forensic imaging method and should be recommended in the investigation of suspicious cardiac deaths.