Generation of a human induced pluripotent stem cell line (SHUPLi002-A) from PBMCs of a healthy female donor.

Human induced pluripotent stem cells (iPSCs) have good multi-lineage differentiation potential, which is an ideal model for studying the pathogenesis of diseases and drug screening.Here, we generated an iPSC line, SHUPLi002-A, from peripheral blood mononuclear cells (PBMCs) of a healthy 28-year-old female individual using episomal vectors. SHUPLi002-A is characterized by expression of classical pluripotent stem cell markers as well as teratoma formation assays to evaluate their differentiation capacity in vivo.

Biomedical application of metal-organic frameworks (MOFs) in cancer therapy: Stimuli-responsive and biomimetic nanocomposites in targeted delivery, phototherapy and diagnosis.

The nanotechnology is an interdisciplinary field that has become a hot topic in cancer therapy. Metal-organic frameworks (MOFs) are porous materials and hybrid composites consisted of organic linkers and metal cations. Despite the wide application of MOFs in other fields, the potential of MOFs for purpose of cancer therapy has been revealed by the recent studies. High surface area and porosity, significant drug loading and encapsulation efficiency are among the benefits of using MOFs in drug delivery. MOFs can deliver genes/drugs with selective targeting of tumor cells that can be achieved through functionalization with ligands. The photosensitizers and photo-responsive nanostructures including carbon dots and gold nanoparticles can be loaded in/on MOFs to cause phototherapy-mediated tumor ablation. The immunogenic cell death induction and increased infiltration of cytotoxic CD8+ and CD4+ T cells can be accelerated by MOF platforms in providing immunotherapy of tumor cells. The stimuli-responsive MOF platforms responsive to pH, redox, enzyme and ion can accelerate release of therapeutics in tumor site. Moreover, MOF nanocomposites can be modified ligands and green polymers to improve their selectivity and biocompatibility for cancer therapy. The application of MOFs for the detection of cancer-related biomarkers can participate in the early diagnosis of patients.

SREBP1 deficiency diminishes glutamate-mediated HT22 cell damage and hippocampal neuronal pyroptosis induced by status epilepticus.

Status epilepticus (SE) is a life-threatening disorder that can result in death or severe brain damage, and there is a substantial body of evidence suggesting a strong association between pyroptosis and SE. Sterol regulatory element binding protein 1 (SREBP1) is a significant transcription factor participating in both lipid homeostasis and glucose metabolism. However, the function of SREBP1 in pyroptosis during SE remains unknown. In this study, we established a SE rat model by intraperitoneal injection of lithium chloride and pilocarpine in vivo. Additionally, we treated HT22 hippocampal cells with glutamate to create neuronal injury models in vitro. Our results demonstrated a significant induction of SREBP1, inflammasomes, and pyroptosis in the hippocampus of SE rats and glutamate-treated HT22 cells. Moreover, we found that SREBP1 is regulated by the mTOR signaling pathway, and inhibiting mTOR signaling contributed to the amelioration of SE-induced hippocampal neuron pyroptosis, accompanied by a reduction in SREBP1 expression. Furthermore, we conducted siRNA-mediated knockdown of SREBP1 in HT22 cells and observed a significant reversal of glutamate-induced cell death, activation of inflammasomes, and pyroptosis. Importantly, our confocal immunofluorescence analysis revealed the co-localization of SREBP1 and NLRP1. In conclusion, our findings suggest that deficiency of SREBP1 attenuates glutamate-induced HT22 cell injury and hippocampal neuronal pyroptosis in rats following SE. Targeting SREBP1 may hold promise as a therapeutic strategy for SE.

Forensic psychiatric analysis of organic personality disorders after craniocerebral injury in Shanghai, China.

Objective: To explore the incidence rate and the differences of clinical manifestations of organic personality disorders with varying degrees of craniocerebral trauma. Materials and methods: According to the International Classification of Diseases-10, 1,027 subjects with craniocerebral trauma caused by traffic accidents were reviewed, the degrees of craniocerebral trauma were graded and those with personality disorder after craniocerebral trauma were diagnosed. The personality characteristics of all patients were evaluated by using the simplified Neuroticism Extraversion Openness Five-Factor Inventory (NEO-FFI). Results: The incidence rate of organic personality disorder after all kinds of craniocerebral trauma was 33.1%, while it was 38.7 and 44.2% in the patients after moderate and severe craniocerebral trauma, respectively, which was significantly higher than that in the patients after mild craniocerebral trauma (18.0%) (P < 0.05). Compared with the patients without personality disorder, the neuroticism, extraversion and agreeableness scores all showed significantly differences (P < 0.05) in the patients with personality disorder after craniocerebral trauma; especially the conscientiousness scores showed significant differences (P < 0.05) in the patients with personality disorder after moderate and severe craniocerebral trauma. The agreeableness and conscientiousness scores in the patients with personality disorder after moderate and severe craniocerebral trauma were significantly lower than that after mild craniocerebral trauma, and the patients with personality disorder after severe craniocerebral trauma had lower scores in extraversion than that after mild craniocerebral trauma. Conclusion: The severity and area of craniocerebral trauma is closely related to the incidence rate of organic personality disorder, and it also affects the clinical manifestations of the latter, which provides a certain significance and help for forensic psychiatric appraisal.

A novel fast-dried urine spot-based method for the analysis of EtS and EtG in urine by liquid chromatography tandem mass spectrometry.

Ethyl sulfate (EtS) and ethyl glucuronide (EtG) in urine are biomarkers to monitor ethanol consumption. Due to their high polarity, severe matrix effects have been observed during analysis of EtS and EtG in urine by liquid chromatography tandem mass spectrometry (LC-MS/MS), which can lead to a loss of sensitivity and accuracy. In the present study, a novel and simple sample preparation approach based on fast-dried urine spot was established to reduce the matrix effect of EtS and EtG in urine. 20 µL of urine was dropped on the Whatman 903# paper and was subsequently dried by microwave in one minute. After ultrasonic assisted extraction with 500 µL of methanol, the analysis was conducted using an LC-MS/MS system. Limits of detection were 5 ng/mL and linear ranges were 10 ng/mL-10 µg/mL for both EtS and EtG. Matrix effects were in the range of 99.3-107.8% for EtS and 86.7-91.0% for EtG at three QC levels. Matrix effects for EtS and EtG were compared between the current method and other sample preparation methods including protein precipitation, and solid-phase extraction. The results showed that this fast-dried urine spot-based extraction method could eliminate matrix effects significantly in analysis of urine EtS and EtG by LC-MS/MS.

Determination of Ethyl Glucuronide and Ethyl Sulfate in Human Whole Blood and Vitreous Humor by LC-MS-MS and Applications to the Interpretation of Postmortem Ethanol Findings.

Identifying the source of ethanol in a decedent remained a complicated problem for forensic toxicologists because of postmortem ethanol formation. As ethanol's non-oxidative metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS) have the potential to distinguish between antemortem ethanol consumption and postmortem ethanol formation, due to their high sensitivity and selectivity. In the current study, a simple and quick liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the determination of EtG and EtS in human whole blood and vitreous humor (VH). A total of 20 µL of the sample was precipitated by methanol, and the analytes were detected by LC-MS/MS in a run of 6 min. This method achieved high sensitivity (limits of detection: 2 ng/mL for both EtG and EtS), with linearity in the range of 5-10,000 ng/mL in both whole blood and VH. Deviations in accuracy, inter- and intra-day precision were all lower than 15% at three quality control levels. Subsequently, this method was applied to 62 real forensic cases. Only blood samples were available in 52 cases. Paired blood and VH samples were present in 10 cases. The concentrations of EtG and EtS in blood were in the range of 0-22,264.8 ng/mL and 0-2,126.0 ng/mL, respectively. In one case with both blood and VH, the blood ethanol concentration was 1.22 mg/mL, with EtG and EtS both below limits of quantification (5 ng/mL) in VH, and no EtG and EtS found in whole blood. The results suggested that EtG and EtS were useful markers for the interpretation of ethanol resource in postmortem blood and VH.

Phosphorylation of PRAS40 contributes to the activation of the PI3K/AKT/mTOR signaling pathway and the inhibition of autophagy following status epilepticus in rats.

Status epilepticus (SE) is a neurological disorder associated with high morbidity and mortality rates, and is often difficult to treat. Moreover, the underlying mechanism of SE remains unknown. The lithium-pilocarpine model is a validated animal model that can reproduce the main clinical and neuropathological features of SE. In the present study, this SE model was utilized and SE was successfully established in rats, as determined by the corresponding epileptic electroencephalogram. Histology, immunohistochemistry, western blot analysis and co-immunoprecipitation were used to detect the phosphorylation (p-) of AKT substrate of 40 kDa (PRAS40), the combination of p-PRAS40 and 14-3-3 protein and the activation of the PI3K/mTOR signaling pathway in SE. In addition, the present study analyzed the dynamics of the expression of autophagy-associated factors in the hippocampus after SE induction, and the influence of suppressing the p- of PRAS40 on the autophagy process was detected in the pathogenesis of SE. The results indicated that increased p-PRAS40 expression could activate the mTOR pathway to decrease the level of autophagy. However, inhibition of the mTOR signaling pathway promoted autophagy flux. These results may provide further understanding of p-PRAS40 functions in SE.

Generation of a human induced pluripotent stem cell line (SHUPLi001-A) from PBMCs of a healthy male donor.

Human induced pluripotent stem cells (iPSCs) are an ideal model for pathomechanism investigation and drug screening due to their differentiation potency. Here, we generated an iPSC line, SHUPLi001-A, from peripheral blood mononuclear cells (PBMCs) of a healthy 30-year-old male individual using non-integrating Sendai virus. Characterization of SHUPLi001-A was confirmed by the expression of typical pluripotency markers and by the teratoma formation assay to assess differentiation potency in vivo.

Forensic appraisal of death due to acute alcohol poisoning: three case reports and a literature review.

Death due to acute alcohol poisoning lacks specific anatomical characteristics, compared with other deaths due to drug poisoning. We report three forensic cases of death from acute alcohol poisoning due to inhibition of the respiratory centre and eventual asphyxia. Blood alcohol concentrations in the three fatalities were 5.28, 3.33 and 3.78 mg/mL, respectively. Lethal doses and blood alcohol concentrations showed differences between individuals. Detailed auxiliary tests besides autopsy were undertaken. These cases show that forensic scientists should exclude other causes of death, combine the autopsy with auxiliary tests, and then make an appraisal.

Incidence of sudden unexpected death in epilepsy in community-based cohort in China.

OBJECTIVE: Sudden unexpected death in epilepsy (SUDEP) is associated with the high premature mortality observed among people with epilepsy. It is, however, considered a rare event in China, probably because of lack of awareness and limitation of studies in the country. We aimed to provide some initial estimation of the burden of SUDEP in China. METHODS: We established a large Chinese community-based cohort of people with epilepsy between January 2010 and December 2011. For any participant who died during follow-up, detailed information on cause of death was obtained using a specifically designed Verbal Autopsy Questionnaire. All cases were reviewed by a multidisciplinary expert panel and reinvestigated if necessary. Sudden unexpected death in epilepsy incidence rates were estimated and case details provided. RESULTS: The cohort consisted of 1562 people and during a median 5years follow-up, 72 deaths were reported. The all-causes death incidence was 11.23 (95% CI 8.86-14.07) per 1000 person-years. Fifteen died suddenly and unexpectedly in a reasonable state of health in the week preceding death. We recorded detailed information of these 15 deaths. Thirteen were considered to be probable SUDEP and two possible SUDEP. The incidence of probable SUDEP was 2.03 (95% CI 1.13-3.38) per 1000 person-years, and the incidence of all suspected (probable and possible) SUDEP was 2.34 (95% CI 1.36-3.77) per 1000 person-years. SIGNIFICANCE: The incidence of SUDEP was relatively high among Chinese people with epilepsy when compared with that in previous community-based studies from high-income countries. The burden of SUDEP in China requires further assessments.

MicroRNA-19b Downregulates Gap Junction Protein Alpha1 and Synergizes with MicroRNA-1 in Viral Myocarditis.

Viral myocarditis (VMC) is a life-threatening disease that leads to heart failure or cardiac arrhythmia. A large number of researches have revealed that mircroRNAs (miRNAs) participate in the pathological processes of VMC. We previously reported that miR-1 repressed the expression of gap junction protein α1 (GJA1) in VMC. In this study, miR-19b was found to be significantly upregulated using the microarray analysis in a mouse model of VMC, and overexpression of miR-19b led to irregular beating pattern in human cardiomyocytes derived from the induced pluripotent stem cells (hiPSCs-CMs). The upregulation of miR-19b was associated with decreased GJA1 in vivo. Furthermore, a miR-19b inhibitor increased, while its mimics suppressed the expression of GJA1 in HL-1 cells. When GJA1 was overexpressed, the miR-19b mimics-mediated irregular beating was reversed in hiPSCs-CMs. In addition, the effect of miR-19b on GJA1 was enhanced by miR-1 in a dose-dependent manner. These data suggest miR-19b contributes to irregular beating through regulation of GJA1 by cooperating with miR-1. Based on the present and our previous studies, it could be indicated that miR-19b and miR-1 might be critically involved in cardiac arrhythmia associated with VMC.

Deregulation of RGS17 Expression Promotes Breast Cancer Progression.

OBJECTIVE: A high level of RGS17 expression is observed in diverse human cancers and correlates with tumor progression. Herein, we aim to investigate its expression and function in breast cancer. METHODS: The expression of RGS17 was detected by immunohistochemical analysis and western blot analysis. The level of miR-32 expression was investigated by qRT-PCR. Western blot analysis was used to determine the relationship between RGS17 and miR-32. A series of loss or gain of function assays was performed to measure the effects of RGS17 or miR-32 on tumor migration, invasion, and proliferation. RESULTS: Compared to that in normal breast specimen, the expression of RGS17 had a significantly higher expression level in breast cancer tissues and cell lines. Although the potential relationship of RGS17 expression with clinicopathological features was not observed, there was a significant correlation of RGS17 expression with p63 expression. In cells, inhibition of RGS17 expression impaired cell migration, invasion, and proliferation. Further, RGS17 was identified as a direct and functional target of miR-32. Overexpression of miR-32 in cells could decrease the expression of RGS17 and inhibit cell migration, invasion, and proliferation. In contrast, ectopic expression of RGS17 could attenuate phenotypes caused by miR-32 overexpression. CONCLUSION: The expression of RGS17 was upregulated in breast cancer, which could enhance cell migration, invasion, and proliferation. Moreover, the RGS17 was identified as a target of miR-32. Our results suggest that RGS17 might play an important role in breast cancer progression and could be a potential target for human breast cancer treatment.

Unnatural deaths in Shanghai from 2000 to 2009: a retrospective study of forensic autopsy cases at the Shanghai Public Security Bureau.

Shanghai is the most developed city in China and has a soaring population. This study uses forensic epidemiology to determine the relationship between unnatural deaths and the development in Shanghai, based on recently released forensic autopsy cases from the 2000s at the Shanghai Public Security Bureau (SPSB). There were 5425 accidental deaths, 2696 homicides, 429 suicides, 186 natural deaths, and 1399 deaths of undetermined cause. There was a male-to-female ratio of 2.02:1, and the average age was 40.9±18.7 years. Traffic accidents (84.2%) were the number one cause of accidental deaths, which decreased during the study period. Sharp force injury (50.6%) was the leading cause of homicides, different from Western countries, where firearms are the leading cause. Hanging (24.5%) was the leading cause of suicides, whereas drug and chemical intoxication was the leading cause in the previous decade; pesticide ingestion decreased in the 2000s. In addition to traffic accidents, manual strangulation was the leading cause of death in childhood fatalities. Children under age 2 were vulnerable to homicides. In the 2000s, there were a large number of drug overdoses, and illegal medical practices and subway-related deaths first appeared in Shanghai. A new type of terrorist attack that involved injecting people with syringes in public places was reflected in the SPSB archives. The forensic epidemiology and changes in unnatural deaths in this decade reflected their relationship with the law, policy and changes in Shanghai. Illegal medical practices, subway-related deaths and terrorist attacks were closely related to the development in Shanghai. Identifying the risks of unnatural deaths will improve public health.

Suicidal drownings with psychiatric disorders in Shanghai: a retrospective study from 2010.1 to 2014.6.

Psychiatric disorders exhibited in 13% suicidal drownings in Southwestern Croatia and 63% in Milan, but in China is unknown. This study is committed to outline the feature of a suicidal drowning with psychiatric disorder, show mental status and reveal key factor to high incidence in China. Immersed corpses were handled by SPSBMPH in its jurisdiction range. Half of immersed corpses were suicidal, and nearly half of suicides had psychiatric disorders. 104 suicidal drownings with psychiatric disorders cases from 2010.1 to 2014.6 were reviewed (21.5% of all immersed corpses, 42.1% of suicides). Most victims clothed normally, and only 2 fastened attached weights. Male victims were more and younger than female. Psycho were prone to commit suicidal drowning in warm and hot season. Psycho were prone to choose familiar area to commit suicide, 45 decedents were found in their familiar areas. Suicidal drowings were occult without suicide attempts, suicide note or abnormal clothing, but showed abnormal mental or behavior changes prior to suicide. The three leading psychiatric disorders were depression (33.7%), depression status (30.8%) and schizophrenia (20.2%). Only 44.2% decedents had visited psychiatric disorder specialist, and merely less than 10% patients could adhere to regular medication. No regular medication on psychiatric disorder was the key factor contributing to high incidence of suicide in psycho. Professional psychiatric and psychological intervention should be taken as soon as possible when they had psychiatric symptoms or suffered misfortune. Guardians should be alert to patients' abnormality to detect their suicidal ideation and intervene, especially in warm season.

JMJD2A-dependent silencing of Sp1 in advanced breast cancer promotes metastasis by downregulation of DIRAS3.

Specificity protein 1(Sp1) is a ubiquitous transcription factor and is highly expressed in breast cancer. However, its expression pattern and role in breast cancer progression remain unclear. The purpose of this study is to examine the expression pattern of Sp1 and determine its role in breast cancer progression. Immunohistochemistry (IHC) was performed on breast cancer tissues to reveal the expression pattern of Sp1. Spearman rank correlation was used for clinical statistics. Gene and protein expressions were monitored by IHC analysis, quantitative polymerase chain reaction, and Western blot. Wound-healing and Transwell assays were conducted to assess the role of Sp1 in breast cancer. Co-immunoprecipitation, deletion mutagenesis, chromatin immunoprecipitation, and dual luciferase reporter gene assays were used for investigation of the regulatory network. Sp1 expression was downregulated in late stage breast cancer and in highly invasive breast cancer cell lines. Expression of Sp1 was negatively correlated with TNM staging (P = 0.002) and metastasis status (P = 0.023). Overexpression of Sp1 inhibited breast cancer cell migratory and invasive abilities, whereas knockdown of GTP-binding RAS-like 3 (DIRAS3, also known as ARHI, NOEY2) attenuated the inhibitory effects. Moreover, re-expression of DIRAS3 abolished Sp1 knockdown-mediated cell migration and invasion. Jumonji domain containing 2A (JMJD2A) inhibited Sp1 autoregulation and explains Sp1 expression pattern in breast cancer. Sp1 negatively regulated breast cancer metastasis by transcriptional activation of DIRAS3. Inhibition of Sp1 autoregulation by JMJD2A contributed to Sp1 expression pattern in breast cancer. Our findings provided evidence that targeted therapy against Sp1 might be useful in early stage breast cancer. However, in late stages, development of Sp1 activator may be more promising for breast cancer treatments.

Epitome of China's unnatural deaths: a historically retrospective study of forensic autopsy cases in Shanghai Public Security Bureau from 1990 to 1999.

The unnatural death investigation in China seems vague to the world. Shanghai is one of the largest city located in Yangtze River Delta in the East China. This study is committed to lift the veil of unnatural death investigation and describe the epitome of China's unnatural deaths. Based on the 7302 forensic report archives from 1990 to 1999 in Shanghai Public Security Bureau, statistics were carried out in 5 areas according to the manner of death. In 3502 accidental deaths, there was a rapid increase during the 1990s, and 71.6% were involved in traffic accidents whose major cause of death was head and neck injuries. The first 3 causes of death in nontraffic accidents (994) were head and neck injuries (42.8%), poisoning (11.8%), and drowning (9.0%). In 2456 homicides, sharp force injury (36.7%), blunt force injury (35.8%), and manual strangulation (12.9%) were the first 3 causes of death. In 563 suicides, drug/chemical intoxication (40.1%), hanging (23.4%), and injuries because of fall from height (11.4%) were the 3 leading causes of death, especially pesticides ingestion. The causes of natural deaths were diseases mainly in circulatory system (23.1%), central nervous system (12.8%), and respiratory system (6.4%). However, the cause of death remained undetermined in 500 victims. Childhood fatalities were different. The victims of accidents and homicides were nearly equal, and the main cause of homicide was manual strangulation. Besides, 1997 was the landmark year when drug abuse began to emerge in Shanghai.

JMJD2A contributes to breast cancer progression through transcriptional repression of the tumor suppressor ARHI.

INTRODUCTION: Breast cancer is a worldwide health problem and the leading cause of cancer death among females. We previously identified Jumonji domain containing 2A (JMJD2A) as a critical mediator of breast cancer proliferation, migration and invasion. We now report that JMJD2A could promote breast cancer progression through transcriptional repression of the tumor suppressor aplasia Ras homolog member I (ARHI). METHODS: Immunohistochemistry was performed to examine protein expressions in 155 cases of breast cancer and 30 non-neoplastic tissues. Spearman correlation analysis was used to analyze the correlation between JMJD2A expression and clinical parameters as well as several tumor regulators in 155 cases of breast cancer. Gene and protein expressions were monitored by quantitative polymerase chain reaction (qPCR) and Western blot. Results from knockdown of JMJD2A, overexpression of JMJD2A, Co-immunoprecipitation (Co-IP) assay, dual luciferase reporter gene assay and chromatin immunoprecipitation (ChIP) elucidated molecular mechanisms of JMJD2A action in breast cancer progression. Furthermore, the effects of ARHI overexpression on JMJD2A-mediated tumor progression were investigated in vitro and in vivo. For in vitro experiments, cell proliferation, wound-healing, migration and invasion were monitored by cell counting, scratch and Boyden Chamber assays. For in vivo experiments, control cells and cells stably expressing JMJD2A alone or together with ARHI were inoculated into mammary fat pads of mice. Tumor volume, tumor weight and metastatic nodules were measured by caliper, electronic balance and nodule counting, respectively. RESULTS: JMJD2A was highly expressed in human breast cancers and positively correlated with tumor progression. Knockdown of JMJD2A increased ARHI expression whereas overexpression of JMJD2A decreased ARHI expression at both protein and mRNA levels. Furthermore, E2Fs and histone deacetylases were involved in the transcriptional repression of ARHI expression by JMJD2A. And the aggressive behavior of JMJD2A in breast cancers could be reversed by re-expression of ARHI in vitro and in vivo. CONCLUSION: We demonstrated a cancer-promoting effect of JMJD2A and defined a novel molecular pathway contributing to JMJD2A-mediated breast cancer progression.