Identification of Key mRNAs and lncRNAs in Neonatal Sepsis by Gene Expression Profiling.

Neonatal sepsis is one of the most prevalent causes of death of the neonates. However, the mechanisms underlying neonatal sepsis remained unclear. The present study identified a total of 1128 upregulated mRNAs and 1008 downregulated mRNAs, 28 upregulated lncRNAs, and 61 downregulated lncRNAs in neonatal sepsis. Then, we constructed PPI networks to identify key regulators in neonatal sepsis, including ITGAM, ITGAX, TLR4, ITGB2, SRC, ELANE, RPLP0, RPS28, RPL26, and RPL27. lncRNA coexpression analysis showed HS.294603, LOC391811, C12ORF47, LOC729021, HS.546375, HNRPA1L-2, LOC158345, and HS.495041 played important roles in the progression of neonatal sepsis. Bioinformatics analysis showed DEGs were involved in the regulation cellular extravasation, acute inflammatory response, macrophage activation of NF-kappa B signaling pathway, TNF signaling pathway, HIF-1 signaling pathway, Toll-like receptor signaling pathway, and ribosome, RNA transport, and spliceosome. lncRNAs were involved in regulating ribosome, T cell receptor signaling pathway, RNA degradation, insulin resistance, ribosome biogenesis in eukaryotes, and hematopoietic cell lineage. We thought this study provided useful information for identifying novel therapeutic markers for neonatal sepsis.

Rapamycin Inhibited Pyroptosis and Reduced the Release of IL-1ß and IL-18 in the Septic Response.

Pyroptosis, an inflammatory form of programmed cell death, is the initiating event of sepsis and results in immune imbalance by releasing IL-1ß and IL-18 in the early stages. Studies show that enhancing autophagy via genetic manipulation can inhibit pyroptosis and prolong the survival of a sepsis animal model, indicating a possible therapeutic strategy against sepsis. However, almost no study so far has achieved pyroptosis inhibition via pharmacological autophagy induction in a sepsis disease model. To this end, we established an in vitro sepsis model by stimulating primary human umbilical vein endothelial cells (HUVECs) with lipopolysaccharide (LPS), and analyzed the effect of the autophagy agonist rapamycin (RAPA) on pyroptosis. Phorbol 12-myristate 13-acetate- (PMA-) activated human THP-1 cells were used as the positive control. LPS significantly increased the levels of the pyroptotic protein Gasdermin D (GSDMD), cysteinyl aspartate-specific proteinase 1 (caspase-1), secreted LDH, IL-1ß, and IL-18. RAPA treatment downregulated the above factors and enhanced autophagy in the LPS-stimulated HUVECs and THP-1 cells. This study shows that RAPA abrogates LPS-mediated increase in IL-1ß and IL-18 by inhibiting pyroptosis and enhancing autophagy.

Umbilical Cord Blood Mesenchymal Stem Cells Enhance Lipopolysaccharide-Induced IL-10 and IL-37 Production in THP-1 Cells.

Umbilical cord blood mesenchymal stem cells (UCB-MSCs) have been shown to be a source of stem cells for use in cellular therapies and have immunomodulatory effects on several immune cells in an inflammatory environment. However, whether UCB-MSCs have immunomodulatory effects against lipopolysaccharide (LPS)-induced inflammatory cytokine secretion in macrophages and whether it is involved in phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway remain unclear. After co-culture of UCB-MSCs and phorbol 12-myristate 13-acetate (PMA)-activated human THP-1 cells using a transwell system, it showed that LPS significantly induced increases in the expression levels of interleukin 10 (IL-10), interleukin 37 (IL-37), phospho-PI3K (p-PI3K), and phospho-Akt (p-Akt) in macrophages. UCB-MSCs upregulated the expression of IL-10, IL-37, p-PI3K, and p-Akt, while it had no obvious effect on PI3K and Akt levels. Inhibitors of PI3K (LY294002) significantly suppressed the expression of IL-10, IL-37, p-PI3K, and p-Akt; however, it had no effect on the expression levels of PI3K and Akt. The present study demonstrated that UCB-MSCs increased the LPS-stimulated expression of IL-10 and IL-37 in macrophages through the PI3K/Akt signaling pathway.

Unexpected custodial death due to acute epiglottitis: A rare autopsy case report.

RATIONALE: Acute epiglottitis is a potentially life-threaten disease, which makes it more challenging to save the life for doctors. Unexpected deaths in custody are a primary cause of concern for the forensic community and doctor worldwide. PATIENT CONCERNS: We present a case of a 44-year-old male detainee who was clinically suspected of dying of acute epiglottitis. The man experienced failure of resuscitation and died after admitted to a hospital. DIAGNOSES: The autopsy, toxicological testing, the test of immunoglobulin E and bacterial culture suggested the patient died of acute epiglottitis. INTERVENTIONS: The bacterial culture was performed to imprecisely identify the cause of death. OUTCOMES: The bacterial culture of the patient's heart blood and nasal and throat swabs showed the presence of the pathogenic microorganism Haemophilus influenza type B. LESSONS: We aim to provide a reference to the medical and forensic community and remind the local law enforcement agencies on the problems present within the correctional healthcare system through this case report. Additionally, we also aim to increase the current knowledge and understanding on custodial deaths caused by natural diseases.

Lessons learned from poisoning cases caused by 2 illegal rodenticides: Tetramine and fluoroacetamide.

BACKGROUND: Tetramine (tetramethylene disulphotetramine, TETS) and fluoroacetamide (FAA) are known as illegal rodenticides with high toxicity to animal species and human beings, which could lead to severe clinical features, including reduction of consciousness, convulsions, coma, and even death. METHODS AND RESULTS: We presented 2 cases that involved rodenticides poisoning. Even though the patients showed severe manifestations, they were initially misdiagnosed, resulting in 2 persons finally died from TETS and FAA poisoning in homicide cases. CONCLUSION: From the clinical and forensic experience of these 2 cases, we suggest that physicians should consider TETS and FAA poisoning when patients present generalized seizures, especially in some cases without clear cause and diagnosis of disease. Early diagnosis and treatment are essential for positive management and criminal investigation in intentional poisoning cases. Moreover, clinical toxicology education should be reinforced.

Child homicide or natural death? A case report of unexpected death of unusual asymptomatic acute laryngotracheobronchitis.

Cases involving the unexpected deaths of children are always a concern for the police and medical examiners alike. In particular, unexpected deaths due to asphyxia without obvious injuries sometimes make decisions regarding the manner of death more difficult. In the present case, a 2-year-old boy was found dead at home, and his mother was initially believed to have killed him. A complete autopsy and forensic investigation were performed, and no injuries were found on the body; however, marked laryngeal edema was observed. Histology showed extensive inflammatory infiltration of the mucosa and submucosa of the larynx, trachea, and bronchi. The cause of death was given as respiratory failure due to acute laryngotracheobronchitis; thus, the manner of death was natural. This case helps to remind the forensic community to keep an open mind and consider a broad differential diagnosis when approaching a case rather than jumping to a conclusion based solely on a preliminary investigation.

Interpolation function estimates post mortem interval under ambient temperature correlating with blood ATP level.

OBJECTIVE: We developed a mathematical model using interpolation function, to characterize the correlation between blood ATP levels in the right ventricle of rabbit and post mortem interval (PMI) at different ambient temperatures. METHODS: Forty-eight healthy rabbits were randomly divided into 6 groups of 8 each. The sacrificed rabbits were maintained in calorstats at 10°C, 15°C, 20°C, 25°C, 30°C and 35°C, respectively. Blood from the right ventricle was sampled every 4h until 72h after death. At different time points, ATP concentrations in the blood samples were measured using an ATP fluorescence rapid detector, and then displayed on the detector screen in the form of relative light units (RLU). Relationship between PMI and ATP degradation levels was investigated statistically by SPSS 17.0 and MATLAB 10.0 software. RESULTS: We obtained six regression equations (Ra(2)=0.887-0.929) with RLU values at PMIs of 72h (10°C), 60h (15°C), 56h (20°C), 52h (25°C), 40h (30°C) and 32h (35°C), and an interpolation function (Ra(2)=0.930) was established with PMI as the dependent variable (z), RLU value as independent variable (x) and temperature as independent variable (y). CONCLUSION: Interpolation function is an appropriate choice for PMI estimation by weakening influence of ambient temperature.

Sudden death in custody due to pituitary apoplexy during long restriction in a sitting position: a case report and review of the literature.

Pituitary apoplexy is an uncommon clinical emergency arising from hemorrhage into or infarction of a pituitary adenoma. The most common presentation is sudden headache, visual field defects and signs of hypopituitarism. It usually occurs in the age group from 20 to 50 years and affects more male than female. Sudden death due to pituitary apoplexy without common symptoms is rarely reported. Here, we described a scarcely-reported case of sudden death in custody caused by pituitary apoplexy resulting from stress-induced hemorrhage of gonadotroph adenoma, a kind of pituitary adenoma, without common clinical symptoms. In this case, a 49-year-old man was restrained in a sitting position for 4 days and died unexpectedly. At autopsy, external examination showed free of trauma. Destruction of bony structure and a circumscribed pituitary tumor were observed in sella turcica. Immunohistochemically, the tumor cells were particular positive for follicle-stimulating hormone (FSH) and luteinizing hormone (LH), thus clarifying the presence of a pituitary gonadotroph adenoma. We provide the case description and a short review of pituitary apoplexy and pituitary adenoma as a rare cause of sudden death.

Fatal extensive bone cement embolism: histological findings confirmed by Fourier transform infrared spectroscopy.

We herein report the first case of fatal extensive bone cement embolism appearing in pulmonary arterioles following surgical vertebral screw augmentation, which histological evidence of bone cement emboli was confirmed by Fourier transform infrared spectroscopy. A 47-year-old woman has accepted multilevel spine fusion and pedicle screw augmentation with totally 4 ml bone cement infusion. She suddenly developed low blood pressure, dyspnoea, and unconsciousness approximately 1 h post anaesthetic recovery, and then she was dead. It was shown both lungs were edematous in autopsy, and bluish emboli were appeared in extensive pulmonary arterioles in H&E stained sections. Negative information was shown in Molybdenum target X-ray imaging, but the emboli were confirmed to be PMMA bone cement by Fourier transform infrared spectroscopy. The present case indicated it might be effective to confirm the dubious trace component in histology by FTIR.

Roles of 3,4-methylenedioxymethamphetamine (MDMA)-induced alteration of connexin43 and intracellular Ca(2+) oscillation in its cardiotoxicity.

UNLABELLED: Although it is well known that 3,4-methylenedioxymethamphetamine (MDMA) can cause various cardiovascular abnormalities and even sudden death from cardiac arrhythmia, whether it has any effect on myocardial gap junctions, which might be one of the targets mediating MDMA-induced cardiotoxicity, remains unclear. OBJECTIVE: To test the hypothesis that MDMA may affect the myocardial gap junction protein connexin43 (Cx43) and induce cardiac dysrhythmia. METHOD: (1) In vivo study: adult rats were treated with a single dose MDMA administration (20mg/kg, i.p.). Electrocardiogram detection and immunohistochemical analysis were performed to evaluate cardiac function and expression of Cx43, respectively; (2) in vitro study: cultured ventricular myocytes of neonatal rats were treated with MDMA (10, 100, 1000µmol/L) for 1h. Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) were performed to investigate the total Cx43 mRNA expression. Immunofluorescent analysis was used to evaluate the amount of junctional Cx43. The phosphorylation status of Cx43 at site Ser368 and intracellular Ca(2+) oscillation were also studied. RESULTS: Obvious changes in electrocardiographic patterns were found in rats following MDMA administration. They were characterized by prolonged QRS duration associated with increased amplitude of QRS complex. The heart rates in treated rats were significantly decreased compared to the rats in the control group. The immunohistochemical findings revealed a significant decrease in Cx43 expression. The in vitro study also showed a marked decline in total Cx43 protein associated with reduction of Cx43 mRNA, whereas the phosphorylated Cx43 at Ser368 was increased. Decrease of junctional Cx43 was found correlated with reduction in N-cadherin induced by high concentration of MDMA. Additionally, confocal microscopy findings revealed alteration of intracellular calcium oscillation patterns characterized by high frequency and increasing influx Ca(2+). CONCLUSIONS: MDMA reduces expression of cardiac gap junction protein Cx43. The increase of phosphorylation status of Cx43 at Ser368 induced by MDMA is attributed, at least in part, to the Ca(2+)-dependent regulation of protein kinase C (PKC) activity. Our findings provide first evidence of MDMA-mediated changes in those cardiac gap junctions that may underlie MDMA-induced cardiac arrhythmia.

Role of neuropeptide Y and peroxisome proliferator-activated receptor γ coactivator-1α in stress cardiomyopathy.

Death following situations of intense emotional stress has been linked to the cardiac pathology described as stress cardiomyopathy, whose pathomechanism is still not clear. In this study, we sought to determine, via an animal model, whether the transcriptional coactivator peroxisome proliferator-activated receptor γ coactivator-1alpha (PGC-1α) and the amino peptide neuropeptide Y (NPY) play a role in the pathogenesis of this cardiac entity. Male Sprague-Dawley rats in the experimental group were subjected to immobilization in a plexy glass box for 1 h, which was followed by low voltage electric foot shock for about 1 h at 10 s intervals in a cage fitted with metallic rods. After 25 days the rats were sacrificed and sections of their hearts were processed. Hematoxylin-eosin staining of cardiac tissues revealed the characteristic cardiac lesions of stress cardiomyopathy such as contraction band necrosis, inflammatory cell infiltration and fibrosis. The semi-quantitative RT-PCR analysis for PGC-1α mRNA expression showed significant overexpression of PGC1-α in the stress-subjected rats (P<0.05). Fluorescence immunohistochemistry revealed a higher production of NPY in the stress-subjected rats as compared to the control rats (P=0.0027). Thus, we are led to conclude that following periods of intense stress, an increased expression of PGC1-α in the heart and an overflow of NPY may lead to stress cardiomyopathy and even death in susceptible victims. Moreover, these markers can be used to identify stress cardiomyopathy as the cause of sudden death in specific cases.

Sudden unexpected death in epilepsy: Evaluation of forensic autopsy cases.

Epilepsy is a common chronic neurological disorder characterized by seizures. Mortality is significantly increased in patients with epilepsy. Sudden unexpected death in epilepsy (SUDEP) is the most common seizure-related category of death. A retrospective study of forensic autopsy cases from 2007 to 2009 at the Office of the Chief Medical Examiner (OCME) yielded a total of 104 sudden unexpected deaths directly or indirectly caused by an epilepsy/seizure disorder in the State of Maryland. Of these deaths, 74 cases met a general accepted definition of SUDEP. The age of SUDEP individuals ranged from 14 to 63 with the majority of subjects in the ages between 21 and 50 years (58 cases, 78.4%). Males were slightly more likely than females to die of SUDEP (male:female=1.5:1 based on the rate). The onset age of epilepsy was documented in 47.3% of cases (35/74) based on investigation and medical records. Of the 35 cases, 12 subjects had early onset epilepsy (onset ages 1-15 years) and 20 subjects had duration of epilepsy for more than 10 years. The majority of deaths (61 of the 74 cases, 82.4%) were unwitnessed. Death scene investigation showed that 71 deaths (95.9%) occurred inside their residence with 50 subjects (70.4%) found either in bed or on the bedroom floor near the bed. Forty-three out of 74 cases (58.1%) showed neuropathological lesions. Per history, 50 subjects were reported as being on anti-epileptic drugs (AEDs). However, postmortem toxicological analysis revealed that only 26 subjects (35.1%) had detectable AEDs. Of the 74 cases, seizure disorder or epilepsy was listed as primary cause of death in 66 cases and the term of SUDEP as official cause of death in only 8 cases. This report focuses on the characteristics of death scene investigation and postmortem examination findings of SUDEP cases.

Involvement of oxidative stress in the mechanism of triptolide-induced acute nephrotoxicity in rats.

Triptolide is one of the most widely used and one of the most potent Chinese traditional herbal medicines. However, side effects, especially nephrotoxicity, limit the use of triptolide. It has been reported that oxidative stress is involved in drug-induced nephrotoxicity. In the present study, we focused on observing triptolide-induced acute nephrotoxicity in rats and investigating whether or not oxidative stress is involved in the pathogenesis of this process. The results showed that a single large dose peritoneal injection of triptolide caused severe oxidative stress characterized by significant decreases of renal SOD and GSH-Px activities, as well as significant increase of renal MDA content and also led to severe impairment of renal structure and function characterized by injury of renal tubules observed in HE-stained and TUNEL-stained slides and increases of Cre and BUN concentrations in a short time. However, pretreatment with the antioxidant vitamin C significantly ameliorated triptolide-induced depletion in renal SOD and GSH-Px activities, caused marked normalization of renal MDA content and also blunt the impairment of renal tubules and renal function. These results suggest that triptolide induces oxidative stress via impairing the antioxidant system, and oxidative stress contributes, at least in part, to the mechanism of triptolide-induced acute nephrotoxicity.

[Comparative analysis of 607 autopsy cases of poisoning death].

OBJECTIVE: To provide references for forensic expertise by investigating the kinds of toxicant, routes of exposure and manners of poisoning deaths, etc. METHODS: Six hundred and seven autopsy cases of poisoning deaths from 1957 to 2008 in Department of Forensic Medicine, Tongji Medical College (Tongji Forensic Science Identification Center of Hubei), were comparatively reviewed. RESULTS: In 218 cases from 1999 to 2008, more than 50% of decedents were male in the ages of 30-49. The toxicants are usually taken orally and the most common manner of death was accidental. The common substances involved in poisoning death were rodenticide, poisoning gas and insecticide. Compared to the data of 1983-1998 and 1957-1982, the common toxic agents had changed significantly. The number of cases involving insecticide and cyanide poisoning decreased in recent years, and the number of cases of rodenticide, poisoning gas, alcohols poisoning displayed an increase tendency, especially for drugs abuse. CONCLUSION: Poisoning deaths of pesticides remain a major public health problem for a long time and the awareness of prevention need to be raised, especially for the prevention of deaths from multiple poisons.

Seven cases of fatal aconite poisoning: forensic experience in China.

This paper presents seven fatal cases of aconite poisoning encountered in the Tongji Center for Medicolegal Expertise in Hubei (TCMEH), China, from 1999 to 2008 retrospectively. In six of the cases, deaths occurred after drinking homemade medicated liquor containing aconite, and in one case death was due to ingestion of traditional Chinese medication containing aconite. Forensic autopsy and pathological examinations ruled out the presence of physical trauma or life-threatening diseases. Diagnosis of aconite poisoning was made after postmortem toxicological analysis. Animal experiment was performed in one case demonstrating that the medicated liquor could cause death rapidly. We present the autopsy and histopathological findings, toxicological analysis, and results of animal experiment done on samples from those seven cases. As an important herbal Chinese medicine, Aconitum species deserve special attention, especially because it contains poisonous alkaloids.

Role of reactive oxygen species in triptolide-induced apoptosis of renal tubular cells and renal injury in rats.

This study investigated the role of reactive oxygen species (ROS) in the pathogenesis of triptolide-induced renal injury in vivo. Rats were randomly divided into 4 groups (n=5 in each): triptolide group in which the rats were intraperitoneally injected with triptolide solution at a dose of 1 mg/kg of body weight on day 8; control group in which the rats received a single intraperitoneal injection of 0.9% physiological saline on day 8; vitamin C group in which the rats were pretreated with vitamin C by gavage at a dose of 250 mg/kg of body weight per day for 7 days before the same treatment as the control group on day 8; triptolide+vitamin C group in which the rats were first subjected to an oral administration of vitamin C at a dose of 250 mg/kg of body weight per day for 7 days, and then to the same treatment as the triptolide group on day 8. All the rats were sacrificed on day 10. Blood samples were collected for detection of plasma creatinine (Pcr) and plasma urea nitrogen (PUN) concentrations. Both kidneys were removed. The histological changes were measured by haematoxylin-eosin (HE) staining. The production of ROS was determined by detecting the fluorescent intensity of the oxidation-sensitive probe rhodamine 123 in renal tissue. Renal malondialdehyde (MDA) content was measured to evaluate lipid peroxidation level in renal tissue. TUNEL staining was performed to assess apoptosis of renal tubular cells. Renal expression of apoptosis-related proteins Bcl-2, Bax, Bid, Bad, Fas and FasL, as well as corresponding encoding genes were assessed by Western Blotting and real-time PCR. The results showed that triptolide treatment promoted the generation of a great amount of ROS, up-regulated the expression of Bax, Bid, Bad, Fas and FasL at both protein and mRNA levels, as well as the ratio of Bax to Bcl-2, and caused the apoptosis of renal tubular cells and renal injury. However, pretreatment with an antioxidant, vitamin C, significantly reduced the generation of ROS and effectively inhibited the triptolide-induced apoptosis of renal tubular cells and renal injury. It was concluded that ROS plays a critical role in triptolide-induced apoptosis of renal tubular cells and renal injury. The protective administration of vitamin C may help alleviate triptolide-induced renal injury and nephrotoxicity.

Death from bilateral pulmonary congenital cystic adenomatoid malformation: a rare case report.

Congenital cystic adenomatoid malformation is one of rare pulmonary hypoplastic diseases. It has been subdivided into 3 types (I-III). Respiratory distress and hydrops are usually diagnosed pre- or postnatally by ultrasonography or radiography. The pathogenesis of CCAM has not been clarified yet. Here, we present an unusual case of bilateral CCAM (type III) of a 2-month-old infant who died suddenly. Concomitant lung malformation and fatty degeneration in hepatic cells make this case unique since such malformation is seldom found in type III CCAM.

Image analysis on corneal opacity: a novel method to estimate postmortem interval in rabbits.

Corneal opacity is one of the most commonly used parameters for estimating postmortem interval (PMI). This paper proposes a new method to study the relationship between changes of corneal opacity and PMI by processing and analyzing cornea images. Corneal regions were extracted from images of rabbits' eyes and described by color-based and texture-based features, which could represent the changes of cornea at different PMI. A KNN classifier was used to reveal the association of image features and PMI. The result of the classification showed that the new method was reliable and effective.

Poisoning deaths in China: type and prevalence detected at the Tongji Forensic Medical Center in Hubei.

The records of 218 poisoning deaths from a Department of Forensic Medicine in a University of China, Tongji Center for Medicolegal Expertise in Hubei (TCMEH), from 1999 to 2008 were retrospectively reviewed. The majority (69.7%) of fatalities was between the ages of 20 and 49 years, and there was a male preponderance (male:female=1.7:1). The most common classes of substances involved in fatalities were rodenticide (19.7%), insecticide and herbicide (17.9%), carbon monoxide (16.5%), drugs (13.8%) and alcohols (12.4%). Ingestion was the predominant route of exposure (65.1%), followed in frequency by inhalation, injection and dermal. In vast majority (64.7%), the manner of death was accidental; suicidal intent was present in 25.2% of cases, homicide in 3.7%, and undetermined 6.4%. When compared to the former reports from the same institution, one for 1956-1984 and another for 1983-1999, an increase was found in the proportion of deaths due to rodenticides, CO, alcohols and drugs, as well as in accidental poisoning deaths. Poisoning deaths due to pesticides remain the major public health problem in China. Further regulatory enforcement should be carried out by government to restrict and manage the use of pesticides and rodenticides which are most toxic to humans.

[Application of confocal laser scanning microscope in forensic pathology].

Confocal laser scanning microscopy(CLSM) is a new technique for microscopic imaging, which can collect the transverse section image of the samples and produce three-dimensional reconstruction and present higher spatial resolution than the conventional light microscope. As a precision instrument for the microscopic image, it plays an important role in forensic pathology. The article reviews the recent research achievements from sudden cardiac death, bullet wound and nervous system damage, etc, and explores the potential applications of the forensic pathology research and forensic practice.