Forensic Study on Objective Evaluation of Visual Acuity of Ametropia with the Event-related Potential P3

Objective In this study,we aimed to assess the characteristics of the P3 component from an event-related potential(ERP)that was induced by visual acuity(VA)processing.Furthermore,we sought to provide electrophysiological evidence for the objective evaluation of VA.Methods We recruited 32 participants with myopia-related ametropia.They reported no other ocular diseases and had an uncorrected VA of 4.0 in both eyes.We used the block letter“E”at different visual angles and orientations as the graphic stimuli.The oddball paradigm,consisting of 4 modules,was used for ERP analysis.The standard stimuli of each module were identical,with a visual angle of 1°15′.The visual angles of the target stimuli were 1°15′,55′,24′,and 15′.The VA test was performed on each eye separately for all participants,and all characteristics of the P3 component were analyzed.Results There was no significant difference in the P3 peak letencies between the target stimulation angle 1°15′group and the 55′group,or between the target stimulation angle 24′group and the 15′group.There was a significant difference in the P3 peak letencies between the target stimulation angle 1°15′group and the 24′group as well as the 15′group.There was a significant difference in the P3 peak letencies between the target stimulation angle 55′group and the 24′group as well as the 15′group.No significant differences were observed in the P3 amplitude between modules.Conclusion In the oddball paradigm,P3 elicitation indicated a cognitive response to the target stimuli.These data showed that the characteristics of P3 can be used as an objective evaluation of VA.

Clinical issues and challenges in imaging of gastrointestinal diseases:A minireview and our experience

Imaging techniques play a crucial role in the modern era of medicine,particularly in gastroenterology.Nowadays,various non-invasive and invasive imaging modalities are being routinely employed to evaluate different gastrointestinal(GI)diseases.However,many instrumental as well as clinical issues are arising in the area of modern GI imaging.This minireview article aims to briefly overview the clinical issues and challenges encountered in imaging GI diseases while highlighting our experience in the field.We also summarize the advances in clinically available diagnostic methods for evaluating different diseases of the GI tract and demonstrate our experience in the area.In conclusion,almost all imaging techniques used in imaging GI diseases can also raise many challenges that necessitate careful consideration and profound expertise in this field.

A NORTHWEST DATABASE MODEL OF SHORT TANDEM REPEAT LOCI IN FORENSIC MEDICINE

Objective To establish the northwest database of short tandem repeat(STR) loci in forensic medicine. Methods Bloodstains or whole blood samples were collected from the unrelated prisoners in Xi'an city. Genetic distribution for 13 STR loci and amelogenin locus were determined in prisons based on GeneScan. One primer for each locus was labeled with the fluorescent by 5 FAM, JOE, or NED. The forensic database were generated by using multiple amplification, GeneScan, genotype, and genetic distribution analysis. Results 113 alleles and 302 genotypes were observed, with the corresponding frequency between 0.0050-0.5250 and 0.0100-0.4100. The mean H was 0.7667. The accumulative DP was 0.9999999,. The accumulative EPP was 0.9999999. The scope of PIC was 0.6036- 0.8562 . PM was less than 10 -11 . The observed and expected genotype frequencies were evaluated using χ 2 test and all were in accordance with Hardy Weinberg equilibrium ( P > 0.05 ). Conclusion STR loci is an ideal genetic marker with powerful polymorphism and stable heredity. It can be used for individual identification and paternity in forensic medicine. The forensic DNA database model can be established successfully.

The value of toxicological analysis in acute poisoning patients with uncertain exposure histories:a retrospective and descriptive study from an institute of poisoning

BACKGROUND:In clinical practice,some patients might not be able or unwilling to provide a thorough history of medication and poison exposure.The aim of this study was to use toxicological analysis to examine the clinical characteristics of patients with acute poisoning whose exposure history was uncertain from a toxicological analysis perspective.METHODS:This was a retrospective and descriptive study from an institute of poisoning.Patient registration information and test reports spanning the period from April 1,2020 to March 31,2022,were obtained.Patients with uncertain exposure histories and who underwent toxicological analysis were included.Clinical manifestations and categories of toxics were analyzed.RESULTS:Among the 195 patients with positive toxicological analysis results,the main causes of uncertain exposure history was disturbance of consciousness(62.6%),unawareness(23.6%)and unwillingness or lack of cooperation(13.8%).The predominant clinical manifestations were disturbed consciousness(62.6%),followed by vomiting and nausea(14.4%)and liver function abnormalities(8.7%).A comparison of clinical manifestations between patients with positive and negative(n=99)toxicological analyses results revealed significantly different proportions of disturbances in consciousness(63%vs.21%),dizziness(1.5%vs.5.1%),multi-organ failure(1.5%vs.7.1%),and local pain(0 vs 4%).The main categories of substances involved were psychiatric medications(23.1%),sedatives(20.5%),insecticides(13.8%),and herbicides(12.8%).CONCLUSION:The clinical manifestations of acute poisoning in patients with an uncertain exposure history are diverse and nonspecific,and toxicological analysis plays a pivotal role in the diagnosis and differential diagnosis of such patients.

Biological,pathological,and multifaceted therapeutic functions of exosomes to target cancer

Exosomes,small tiny vesicle contains a large number of intracellular particles that employ to cause various diseases and prevent several pathological events as well in the human body.It is considered a“double-edged sword”,and depending on its biological source,the action of exosomes varies under physiological conditions.Also,the isolation and characterization of the exosomes should be performed accurately and the methodology also will vary depending on the exosome source.Moreover,the uptake of exosomes from the recipients’cells is a vital and initial step for all the physiological actions.There are different mechanisms present in the exosomes’cellular uptake to deliver their cargo to acceptor cells.Once the exosomal uptake takes place,it releases the intracellular particles that leads to activate the physiological response.Even though exosomes have lavish functions,there are some challenges associated with every step of their preparation to bring potential therapeutic efficacy.So,overcoming the pitfalls would give a desired quantity of exosomes with high purity.

Characteristics of Pattern Visual Evoked Potential in Two Eyes with Varying Visual Acuity in One Eye and Forensic Application

In this study, we aimed to study the pattern visual evoked potentials （P-VEPs） in two eyes with varying visual acuity in one eye and to provide an objective estimation of visual acuity by comparing P-VEPs in one and two eyes. Thirty subjects were chosen, who had one eye with an acuity of 5.0, 4.85, 4.6, 4.0, or scieropia and obstructed vision and the other eye with an acuity of 5.0, respectively. P-VEPs were detected under the large grating stimuli at 3x4 spatial frequency, moderate grating stimuli （12× 16 spatial frequency） and small grating stimuli （48×64 spatial frequency）. Under large grating stimuli, there was no significant difference in P100 peak latency between the groups, nor was there a significant difference between the amplitude of two eyes and the amplitude of one normal-vision eye. Under moderate and small grating stimuli, there was a significant difference in P100 peak latency between the group with both eyes having an acuity of 5.0 and the group with visual acuity below 4.0 in one eye. There was a significant difference in P100 amplitude between the group with visual acuity of 5.0 in both eyes and the group with one normal-vision eye. There was no significant difference in the amplitude of two eyes and the amplitude of one normal-vision eye between any other two groups. In forensic identification, characteristics and variability of P-VEPs in one and two eyes can be used to identify malingering or decline in visual acuity.

Experimental analysis of ballistic trauma in a human body protected with 30 layer packages made of biaxial and triaxial Kevlar^(■)29 fabrics

Behind armour blunt trauma(BABT)is a body injury resulting from the deformation of the back surface of armour as a result of a bullet impact.In the case of textile body armour,the severity of the injury may depend on the material of the fibres,but also on the geometric structure of the fabric.The article focuses on experimental research into injuries of the human body protected by ballistic packets made of biaxial and triaxial fabrics,during a non-penetrating impact from a Parabellum 9 mm×19 mm Full Metal Jacket(FMJ)bullet,at a speed of 406±5 m/s.In experimental research,the fabrics had a comparable surface weight and were made of the same Kevlar^(■)29 yarn.The ballistic packages were made of 30 layers.As part of the work,a physical model of the human body was developed.The human body model consisted of a model of the heart,lungs,and skeletal and muscular systems.During the bullet impact,the pressure forces were recorded using sensors located at selected points of the human body.The bullets hit five selected places on the body that were considered critical,from the point of view of maintaining a human’s vital functions.It was found that,during firing,pressure increases both at the site of impact and in the internal organs,which can lead to multi-organ damage.As a result of the experimental analysis,it has been shown that the pressures exerted on specific organs are always lower in the case of body protection with a ballistic packet made of triaxial fabrics,compared to a packet made of biaxial fabrics.

Association of Five SNPs at the PARK16 locus as a Suscepti- bility Locus with Parkinson＇s Disease for Forensic Application

To investigate the association of five SNPs(rs823083,rs708723,rs4951261,rs823076 and rs16856110) at the PARK16 locus with Parkinson's disease(PD),and to potentiate its forensic application.The genomic DNAs of 215 PD patients and 212 matched controls from the northern Han Chinese population were amplified in two independent PCR systems and subsequently genotyped by digestion with the three endonucleases(Hinf Ⅰ,Nco Ⅰ and Msp Ⅰ).The genetic parameters and association studies were carried out with SPSS 13.0,Haploview version 4.2 and PLINK 1.07 softwares.We detected accurately all genotypes in the five SNPs with multiplex PCR-RFLP and mismatched multiplex PCR-RFLP techniques.The genotypes of four SNPs,except for rs823083,were in Hardy-Weinberg equilibrium.The four SNPs,rs16856110,rs4951261,rs708723 and rs823076,which were in linkage equilibrium,should not be associated with PD(P-values ranging from 0.077 to 0.544).The SNPs investigated at the PARK16 locus were not found to be involved in PD-associated blocks in the northern Han Chinese population.The allele distributions of rs708723,rs4951261,rs823076 and rs16856110 in the northern Han Chinese population can be highly polymorphic,which can be applied to genetic analysis and forensic practices.

Improving DNA Data Capacity:Forensic Parameters and Genetic Structure Analysis of Jinjiang Han Population with the Microreader^(TM) Y Prime Plus ID System

Objective Population genetic analysis based on genetic markers harbors valuable forensic applications.In this regard,it is informative and imperative to explore Han groups as they are the largest population of China.In particular,there is a largely underrepresented amount of information from recent decades regarding the southeast costal Han Chinese.Therefore,the aim of this study is to investigate the available genetic characteristics of the Han population living in the Jinjiang,Fujian Province,Southeastern China.Methods We sampled 858 saliva samples and used the commercially available Microreader^(TM) Y Prime Plus ID System to identify population data of Y-short tandem repeat(STR)loci of this region.Results A total of 822 different haplotypes were observed.The overall haplotype diversity,discriminatory power and haplotype match probability were 0.9999,0.9999 and 0.0012,respectively.Conclusion Our results showed that the Jinjiang Han population was closely genetically related to Han groups of China.Overall,we identified a set of 37 Y-STRs that are highly polymorphic,and that can provide meaningful information in forensic practice and human genetic research.

The combination of NlMIP and Gαi/q coupled-receptor NlA10promotes abdominal vibration production in female Nilaparvata lugens(Stal)

For various sexually mature insects,including the brown planthopper(BPH,Nilaparvata lugens),the abdominal vibration(AV)signal is the initiation of the mating process,and itiscritical to the successofmating.Currently,there are few studies on the genetic and molecular mechanisms of AV regulation.Ourprevious AV-related transcriptomic study in female BPH identified myoinhibitory peptide(NlMIP)as a gene that potentially affects AV statusinfemales,buthow NlMIP affects AV status remains unknown.In this study,we confirmed that NlMIP regulates AV production and mating behavior in female BPH.When the RNAi knockdown efficiency of NlMIP was 59.00%,the probability of females producing AV and the mating rate in 1 h decreased by 38.89 and 61.11%,respectively.In addition,six mature peptides of NlMIP were synthesized and they were able to regulate AV production and mating behavior in females,with NlMIP2having the strongest effect.The A-family neuropeptide GPCR 10(NlA10)was found to be a potential receptor for NlMIP based on a phylogenetic tree analysis and the fact that NlMIP mature peptides effectively activated NlA10.After NlA10was knocked down,the probability of females producing AV and the mating rate in 1 h had reductions of 28.89 and 43.33%,respectively.When activated by NlMIP2,NlA10 coupled the Gαi/q signalling pathways,thereby inhibiting the downstream AC/cAMP/PKA,activating the PLC/Ca^(2+)/PKC signalling pathways and then activating MEK1/2 in a cascade to mediate the phosphorylation of ERK1/2,and finally regulating the AV of females.These results provide a basis for the prevention and control of the brown planthopper pest by disrupting female AV.

Identification of Mixtures of Two Types of Body Fluids Using the Multiplex Methylation System and Random Forest Models

Objective Body fluid mixtures are complex biological samples that frequently occur in crime scenes,and can provide important clues for criminal case analysis.DNA methylation assay has been applied in the identification of human body fluids,and has exhibited excellent performance in predicting single-source body fluids.The present study aims to develop a methylation SNaPshot multiplex system for body fluid identification,and accurately predict the mixture samples.In addition,the value of DNA methylation in the prediction of body fluid mixtures was further explored.Methods In the present study,420 samples of body fluid mixtures and 250 samples of single body fluids were tested using an optimized multiplex methylation system.Each kind of body fluid sample presented the specific methylation profiles of the 10 markers.Results Significant differences in methylation levels were observed between the mixtures and single body fluids.For all kinds of mixtures,the Spearman’s correlation analysis revealed a significantly strong correlation between the methylation levels and component proportions(1:20,1:10,1:5,1:1,5:1,10:1 and 20:1).Two random forest classification models were trained for the prediction of mixture types and the prediction of the mixture proportion of 2 components,based on the methylation levels of 10 markers.For the mixture prediction,Model-1 presented outstanding prediction accuracy,which reached up to 99.3%in 427 training samples,and had a remarkable accuracy of 100%in 243 independent test samples.For the mixture proportion prediction,Model-2 demonstrated an excellent accuracy of 98.8%in 252 training samples,and 98.2%in 168 independent test samples.The total prediction accuracy reached 99.3%for body fluid mixtures and 98.6%for the mixture proportions.Conclusion These results indicate the excellent capability and powerful value of the multiplex methylation system in the identification of forensic body fluid mixtures.

Haplotype and Genetic Analysis of 41 Y-STR Loci in the Wuwei Han Population from Gansu Province,China

Objective Y-Chromosomal short tandem repeat polymorphism(Y-STR)analysis plays an indispensable role in the identification of male individuals,population genetics,and biogeographic research.While profiles of many populations based on Y-STR markers in human genomes are ample,haplotype data for the Wuwei Han are still scarce.Methods In this study,2180 unrelated Wuwei Han male individuals residing in Gansu Province,China were collected and genotyped using the novel Microreader™40Y Plus ID system.Phylogenetic relationship reconstructions,multidimensional scaling(MDS),and heatmap analysis were performed based on the genetic distance(Rst)values between our studied population and other populations of the Ymax module in the Y-STR Haplotype Reference Database(YHRD).Results A total of 2129 unique haplotypes were obtained,and the haplotype diversity(HD)and discrimination capacity(DC)for the Wuwei Han were 0.9999 and 0.9931,respectively.Conclusion Our results demonstrate that the Wuwei Han population had intimate genetic relationships with East Asians,especially the geographically close Han populations.Overall,this Y-Chromosomal assay gives valuable information about paternal lineages in male individual tracking and genealogical database construction.

Research Progress on the Association between Schizophrenia and Toxoplasma gondii Infection

Toxoplasma gondii(T.gondii or Tg),is an obligatory intracellular parasite with humans as its intermediate hosts.In recent years,significant correlations between T.gondii infection and schizophrenia have been reported,including the possible mediating mechanisms.Currently,mechanisms and hypotheses focus on central neurotransmitters,immunity,neuroinflammation,and epigenetics;however,the exact underlying mechanisms remain unclear.In this article,we review the studies related to T.gondii infection and schizophrenia,particularly the latest research progress.Research on dopamine(DA)and other neurotransmitters,the blood-brain barrier,inflammatory factors,disease heterogeneity,and other confounders is also discussed.In addition,we also summarized the results of some new epidemiological investigations.

Improving Evidence-Based Practice in Forensic Medicine:Validation of INtegration of Forensic Epidemiology and the Rigorous EvaluatioN of Causation Elements and Principles of Evidence-Based Reporting in FORensic Medicine-Pathology version in Indonesia

Introduction: Expert opinions presented in legal proceedings should be scientifically accountable, which is known as evidence-based practice (EBP). Although forensic medical expert opinions are essential in legal proceedings, the methods used to formulate them are not always evidence based or based on standard methods. In forensic medicine, EBP has not been explicitly applied, including in Indonesia. One potential approach to formulate evidence-based expert opinions is called INtegration of Forensic Epidemiology and the Rigorous EvaluatioN of Causation Elements (INFERENCE). In addition, there is also no universal guideline for making forensic pathological reports. One prospective guideline is named the Principles of Evidence-based Reporting in FORensic Medicine-Pathology version (PERFORM-P). Methods: This article describes the validation process of INFERENCE and PERFORM-P in Indonesia. This study uses a mixed method through three interrelated phases, i.e., (1) a cross-sectional survey to determine the characteristics of Indonesian forensic doctors and their current practice, (2) the adaptation and validation process of the two tools through a review by the Indonesian College of Forensic Medicine, and (3) a one-group pre-postintervention study to assess the validity and reliability of forensic medical expert opinions formulated using Indonesian-INFERENCE (i-INFERENCE) and reported using Indonesian-PERFORM-P (i-PERFORM-P). Results and Discussion: In general, both tools received a positive reception and can potentially be used in the Indonesian setting with some additions/clarifications in the user manuals. Participants envision that both tools will be most useful in complex cases. Conclusions: By obtaining the i-INFERENCE and the i-PERFORM-P, it is hoped that Indonesian forensic medical doctors are better equipped in analyzing and reporting complex cases, and the implementation of EBP can be improved.

Early expressions of hypoxia-inducible factor 1alpha and vascular endothelial growth factor increase the neuronal plasticity of activated endogenous neural stem cells after focal cerebral ischemia

Endogenous neural stem cells become ＂activated＂ after neuronal injury, but the activation sequence and fate of endogenous neural stem cells in focal cerebral ischemia model are little known. We evaluated the relationships between neural stem cells and hypoxia-inducible factor-1α and vascular endothelial growth factor expression in a photothromobotic rat stroke model using immunohistochemistry and western blot analysis. We also evaluated the chronological changes of neural stem cells by 5-bromo-2′-deoxyuridine（BrdU） incorporation. Hypoxia-inducible factor-1α expression was initially increased from 1 hour after ischemic injury, followed by vascular endothelial growth factor expression. Hypoxia-inducible factor-1α immunoreactivity was detected in the ipsilateral cortical neurons of the infarct core and peri-infarct area. Vascular endothelial growth factor immunoreactivity was detected in bilateral cortex, but ipsilateral cortex staining intensity and numbers were greater than the contralateral cortex. Vascular endothelial growth factor immunoreactive cells were easily found along the peri-infarct area 12 hours after focal cerebral ischemia. The expression of nestin increased throughout the microvasculature in the ischemic core and the peri-infarct area in all experimental rats after 24 hours of ischemic injury. Nestin immunoreactivity increased in the subventricular zone during 12 hours to 3 days, and prominently increased in the ipsilateral cortex between 3–7 days. Nestin-labeled cells showed dual differentiation with microvessels near the infarct core and reactive astrocytes in the peri-infarct area. BrdU-labeled cells were increased gradually from day 1 in the ipsilateral subventricular zone and cortex, and numerous BrdU-labeled cells were observed in the peri-infarct area and non-lesioned cortex at 3 days. BrdU-labeled cells rather than neurons, were mainly co-labeled with nestin and GFAP. Early expressions of hypoxia-inducible factor-1α and vascular endothelial growth factor after ischemia made up the microenvironment to increase the neuronal plasticity of activated endogenous neural stem cells. Moreover, neural precursor cells after large-scale cortical injury could be recruited from the cortex nearby infarct core and subventricular zone.

Expression of macrophage migration inhibitory factor is associated with enhanced angiogenesis and advanced stage in gastric carcinomas

AIM: Macrophage migration inhibitory factor (MIF) was reported to inactivate p53 and play an essential role in the growth and angiogenesis of tumors that arise at sites of chronic inflammation. Gastric inflammation is a prerequisite for the development of gastric carcinoma (GC), which has recently been linked to Helicobacter pylori (H pylori) infection. This study aimed to investigate dinicopathological significance of MIF expression in GCs. METHODS: We selected 90 consecutive patients with GCs for investigation of the relation among MIF status, clinicopathological parameters, p53 expression and angiogenesis. MIF and p53 expression was assessed by immunohistochemistry as positive and negative groups. Tumor vascularity was evaluated by counting microvessel density on anti-CD34 stained sections. Expression status of MIF was correlated with determined dinicopathological data, p53 immunoreactivity and microvessel counts. RESULTS: Strong immunostainings of MIF were observed in the cytoplasm of cancerous cells in 40% (36/90) of cases but not in normal or metaplastic epithelia. There was no statistically significant correlation between MIF expression and age, gender, H pylori infection, tumor location, histological subtypes, lymph node metastasis or p53 expression. Early GC less frequently overexpressed MIF as compared to advanced GCs (4/20 vs 32/70, P= 0.04). A remarkably increased microvessel count was noted in GCs with MIF expression than those without MIF expression (55.1±30.1 vs 31.3±28.8, P= 0.0001). CONCLUSION: Our results suggest that expression of MIF may contribute to the progression and enhanced angiogenesis in a substantial portion of GCs.

Insights on augmenter of liver regeneration cloning and function

Hepatic stimulator substance （HSS） has been referred to as a liver-specific but species non-specific growth factor. Gradient purification and sequence analysis of HSS protein indicated that it contained the augmenter of liver regeneration （ALR）, also known as hepatopoietin （HPO）. ALR, acting as a hepatotrophic growth factor, specifically stimulated proliferation of cultured hepatocytes as well as hepatoma cells in vitro, promoted liver regeneration and recovery of damaged hepatocytes and rescued acute hepatic failure in vivo. ALR belongs to the new Erv1/Alr protein family, members of which are found in lower and higher eukaryotes from yeast to man and even in some double-stranded DNA viruses. The present review article focuses on the molecular biology of ALR, examining the ALR gene and its expression from yeast to man and the biological function of ALR protein. ALR protein seems to be non-liver-specific as was previously believed, increasing the necessity to extend research on mammalian ALR protein in different tissues, organs and developmental stages in conditions of normal and abnormal cellular growth.

Expressions of Osteopontin (OPN),αVβ3 and Pim-1 Associated with Poor Prognosis in Non-small Cell Lung Cancer (NSCLC)

Objective： To examine the expressions ot osteopontin （OPN）, αvP3 and Pim-1 in non-small cell lung cancer （NSCLC）, and investigate their potential pathogenic roles in the development of NSCLC. Methods： Immunohistochemistry was used to examine the expressions of OPN, αve3 and Pim-1 in cohort （136 cases） of NSCLC samples and their adjacent normal lung tissue specimens. Statistical analysis was performed to evaluate the relationships among expressions of OPN, αve3 and Pim-1 and their associations with patients clinico- pathological parameters. Results： The expressions of OPN and Pim-1 were predominantly observed in cytoplasm. The expression of αve3 was mostly detected in cytoplasm and/or membrane. In NSCLC samples, the positive rates of OPN, αve3 and Pim-1 expressions were 68.4% （93/136）, 77.2% （105/136） and 57.4% （78/136）, respectively. In normal lung tissues, in contrast, the positive rates of OPN, αve3 and Pim-1 were 24.0% （12/50）, 26.0% （13/50） and 16.0% （8/50）, respectively. There were significant differences of the positive expression rates of OPN, αve3 and Pim-1 between NSCLCs samples and normal lung tissues （P〈O.01）. In addition, the positive expression of OPN, αve3 and Pim-1 in NSCLCs samples was significantly associated with increased pathological grade, lymph node metastasis and advanced clinical stage （P〈O.01）, and they were independent of other clinicopathological parameters （P〉0.05）. Furthermore, a significantly positive correlation between the expression of OPN and αve3 （r=0.38, P〈O.O1）, OPN and Pim-1 （r=0.37, P〈O.01）, or av133 and Pim-1 （r=0.20, P〈0.05） was evaluated in our NSCLC cohort. Conclusion： OPN, αve3 and Pim-1 proteins are frequently overexpressed in NSCLC, and they may play important roles in the development and/or progression of NSCLC.

Comparison of labeling methods and time course of traumatic brain injury-induced cell death in mice

BACKGROUND：Various molecular mechanisms of cell death following traumatic brain injury have been previously described.However,the time course of cell death remains unclear.TUNEL and Fluoro-Jade B labeling have been widely used to label apoptotic cells and neuronal degeneration.Propidium iodide （PI） functions as a biomarker of cell death in vivo.OBJECTIVE：To explore the role of PI labeling compared to TUNEL and Fluoro-Jade B staining for detecting neural cell death,and to observe time course of traumatic brain injury-induced cell death in mice.DESIGN,TIME AND SETTING：A randomized,controlled,animal experiment was performed at the Laboratory of Aging and Nervous Diseases,Soochow University from September 2007 to December 2008.MATERIALS：PI （B1221） was purchased from Sigma,USA.TUNEL kit was purchased from Roche Molecular Biochemicals,USA.Fluoro-Jade B was purchased from Chemicon,USA.METHODS：A total of 70 healthy,male,Kunming mice were randomly assigned to sham-surgery （n = 5） and model （n = 65） groups.Traumatic brain injury was established using the controlled cortical impact method.PI was intraperitoneally injected at 1 hour prior to animal sacrifice.MAIN OUTCOME MEASURES：TUNEL,Fluoro-Jade B,and Pl-positive cells were quantified using a double-labeling method to determine the time course of traumatic brain injury-induced cell death.RESULTS：PI labeled cells in an earlier phase of cell death than TUNEL and Fluoro-Jade B labeling.Pl-positive cells were observed immediately following injury,and the numbers rapidly increased in injured brain areas at 1 hour,peaked at 24-48 hours,and subsequently decreased at 3-21 days post-injury.TUNEL-labeled cells were significantly increased at 12 hours,while Fluoro-Jade B-labeled cells were increased at 6 hours after injury,with cells still visible at 6-48 hours post-injury.Moreover,a greater number of Pl-positive cells were observed compared to TUNEL- and Fluoro-Jade B-labeled cells.CONCLUSION：PI labeling is more sensitive and reliable than TUNEL and Fluoro-Jade B staining for detecting cell death following traumatic brain injury.Moreover,PI labeling can function as a reliable marker to estimate the entire time course of cell death.

A safety evaluation of profound hypothermia-induced suspended animation for delayed resuscitation at 90 or 120min

Background: The successful treatment of military combat casualties with penetrating injuries is significantly dependent on the time needed to get the patient to an adequate treatment facility. Profound hypothermia induced suspended animation for delayed resuscitation(SADR) is a novel approach for inducing cardiac arrest and buying additional time for such injuries. However, the time used to safely administer circulatory arrest(CA) is controversial. The goal of this study was to evaluate the safety of hypothermia-induced SADR over 90 and 120 min time intervals.Methods: Sixteen male BAMA minipigs were randomized into two groups: CA90 group(90 min, n =8) and CA120 group(120 min, n =8). Cannulation of the right common carotid arteries and internal jugular veins was performed to establish cardiopulmonary bypass for each animal. Through the perfusion of cold organ preservation solution(OPS), cardioplegia and profound hypothermia(15℃) were induced. After CA, cardiopumonary bypass(CPB) was restarted, and the animals were gradually re-warmed and resuscitated. The animals were assisted with ventilators until spontaneous breathing was achieved. The index of hemodynamic perioperative serum chemistry values [alanine transaminase(ALT), aspartate aminotransferase(AST), creatinine(CR), lactic dehydrogenase(LDH) and troponin T(TnT)] and survival were observed from pre-operation to 7 days post-operation.Results: Fifteen animals were enrolled in the experiment, while 1 animal in CA120 group died from surgical error. All 8 animals in CA90 group recovered, with only 1 animal displaying mild disability. However, in CA120 group, only 2 animals survived with severe disability, and the other 5 animals died after 2 days post-operation. In CA90 group, the perioperative serum chemistry values increased at 1 day post-operation(ALT 84.43±18.65 U/L; AST 88.99±23.19 U/L; Cr 87.90±24.49μmol/L; LDH 1894.13±322.26 U/L; TnT 0.849±0.135 ng/ml) but decreased to normal or almost normal levels at 7 days post-operation(ALT 52.48±9.04 U/L; AST 75.23±21.46 U/L; Cr 82.69±18.41μmol/L; LDH 944.67±834.32 U/L; TnT 0.336±0.076 ng/ml).Conclusion: Profound hypothermia-induced SADR is an effective method for inducing cardiac arrest. Our results indicate that inducing CA for 90 min(at 15℃) is safer than doing so for 120 min. Our results indicate that 120 min of CA at 15℃ is dangerous and can result in high mortality and severe neurological complications. Further experimentation is needed to determine whether 120 min of CA at temperatures lower than 15℃ can lead to safe recovery.