Caspase-12 is Expressed in Purkinje Neurons and Prevents Psychiatric-Like Behavior in Mice.

Caspase-12 is a caspase family member for which functions in regulating cell death and inflammation have previously been suggested. In this study, we used caspase-12 lacZ reporter mice to elucidate the expression pattern of caspase-12 in order to obtain an idea about its possible in vivo function. Strikingly, these reporter mice showed that caspase-12 is expressed explicitly in Purkinje neurons of the cerebellum. As this observation suggested a function for caspase-12 in Purkinje neurons, we analyzed the brain and behavior of caspase-12 deficient mice in detail. Extensive histological analyses showed that caspase-12 was not crucial for establishing cerebellum structure or for maintaining Purkinje cell numbers. We then performed behavioral tests to investigate whether caspase-12 deficiency affects memory, motor, and psychiatric functions in mice. Interestingly, while the absence of caspase-12 did not affect memory and motor function, caspase-12 deficient mice showed depression and hyperactivity tendencies, together resembling manic behavior. Next, suggesting a possible molecular mechanistic explanation, we showed that caspase-12 deficient cerebella harbored diminished signaling through the brain-derived neurotrophic factor/tyrosine kinase receptor B/cyclic-AMP response binding protein axis, as well as strongly enhanced expression of the neuronal activity marker c-Fos. Thus, our study establishes caspase-12 expression in mouse Purkinje neurons and opens novel avenues of research to investigate the role of caspase-12 in regulating psychiatric behavior.

Remote Ischemic Postconditioning-Mediated Neuroprotection against Stroke by Promoting Ketone Body-Induced Ferroptosis Inhibition.

Neuronal death resulting from ischemic stroke is the primary cause of adult mortality and disability, and effective neuroprotective agents for poststroke intervention are still lacking. Remote ischemic postconditioning (RIPostC) has demonstrated significant protective effects against ischemia in various organs; however, the specific mechanisms are not fully understood. This study investigated the potential neuroprotective mechanisms of RIPostC in the context of ischemic stroke. Using a rat model of middle cerebral artery occlusion, we found that RIPostC mitigated neurological damage, improved movement in the open-field test, and protected against neuronal apoptosis. In terms of energy metabolism, RIPostC enhanced ATP levels, suppressed lactate content, and increased the production of ketone bodies (KBs). In the ferroptosis assay, RIPostC protected against lipoperoxidation, reversed the reduction of glutathione peroxidase 4 (GPX4), and mitigated the excessive expression of long-chain acyl-CoA synthetase family member 4 (ACSL4). In oxygen-glucose deprivation/reoxygenation-treated HT22 cells, KBs maintained GPX4 levels, suppressed ACSL4 expression, and preserved the mitochondrial cristae number. However, the effect of KBs on the expression of GPX4, ACSL4, and the number of mitochondrial cristae was blocked by erastin. Moreover, both RIPostC and KBs reduced total iron and ferrous ion content by repressing iron transporters both in vitro and in vivo. In conclusion, KBs-induced mitigation of ferroptosis could represent a new therapeutic mechanism for RIPostC in treating stroke.

Causal Association Between Tea Consumption and Gout: A Mendelian Randomization Study.

OBJECTIVE: Evidence from prospective studies on the consumption of tea and risk of gout is conflicting and limited. We aimed to investigate the potential causal effects of tea intake on gout using Mendelian randomization (MR). METHODS: Genome-wide association studies in UK Biobank included 349 376 individuals and successfully discovered single-nucleotide polymorphisms linked to consumption of one cup of tea per day. Summary statistics from the Chronic Kidney Disease Genetics consortium included 13 179 cases and 750 634 controls for gout. Two-sample MR analyses were used to evaluate the relationship between tea consumption and gout risk. The inverse-variance weighted (IVW) method was used for primary analysis, and sensitivity analyses were also conducted to validate the potential causal effect. RESULTS: In this study, the genetically predicted increase in tea consumption per cup was associated with a lower risk of gout in the IVW method (OR: 0.90; 95% CI: 0.82-0.98). Similar results were found in weighted median methods (OR: 0.88; 95% CI: 0.78-1.00), while no significant associations were found in MR-Egger (OR: 0.89; 95% CI: 0.71-1.11), weighted mode (OR: 0.80; 95% CI: 0.65-0.99), and simple mode (OR: 1.01; 95% CI: 0.75-1.36). In addition, no evidence of pleiotropy was detected by MR-Egger regression (P=0.95) or MR-PRESSO analysis (P=0.07). CONCLUSION: This study provides evidence for the daily consumption of an extra cup of tea to reduce the risk of gout.

Protein-Scaffolded DNA Nanostructures for Imaging of Apurinic/Apyrimidinic Endonuclease 1 Activity in Live Cells.

Nucleic acids are valuable tools for intracellular biomarker detection and gene regulation. Here we propose a new type of protein (avidin)-scaffolded DNA nanostructure (ADN) for imaging the activity of apurinic/apyrimidinic endonuclease 1 (APE1) in live cells. ADN is designed by assembling an avidin-displayed abasic site containing DNA strands labeled with a fluorophore or a quencher via a complementary linker strand. ADN is nonemissive due to the close proximity of fluorophores and quenchers. APE1-mediated cleavage separates the fluorophores from the quenchers, delivering activated fluorescence. In vitro assays show that ADN is responsive to APE1 with high sensitivity and high specificity. ADN can efficiently enter the cells, and its capability to visualize and detect intracellular APE1 activities is demonstrated in drug-treated cells and different cell lines. The modular and easy preparation of our nanostructures would afford a valuable platform for imaging and detecting APE1 activities in live cells.

Crosstalk between PML and p53 in response to TGF-ß1: A new mechanism of cardiac fibroblast activation.

Cardiac fibrosis is a common pathological cardiac remodeling in a variety of heart diseases, characterized by the activation of cardiac fibroblasts. Our previous study uncovered that promyelocytic leukemia protein (PML)-associated SUMO processes is a new regulator of cardiac hypertrophy and heart failure. The present study aimed to explore the role of PML in cardiac fibroblasts activation. Here we found that PML is significantly upregulated in cardiac fibrotic tissue and activated cardiac fibroblasts treated with transforming growth factor-ß1 (TGF-ß1). Gain- and loss-of-function experiments showed that PML impacted cardiac fibroblasts activation after TGF-ß1 treatment. Further study demonstrated that p53 acts as the transcriptional regulator of PML, and participated in TGF-ß1 induced the increase of PML expression and PML nuclear bodies (PML-NBs) formation. Knockdown or pharmacological inhibition of p53 produced inhibitory effects on the activation of cardiac fibroblasts. We further found that PML also may stabilize p53 through inhibiting its ubiquitin-mediated proteasomal degradation in cardiac fibroblasts. Collectively, this study suggests that PML crosstalk with p53 regulates cardiac fibroblasts activation, which provides a novel therapeutic strategy for cardiac fibrosis.

Metabolomic Profiling of Brain Protective Effect of Edaravone on Cerebral Ischemia-Reperfusion Injury in Mice.

Edaravone (EDA) injection has been extensively applied in clinics for treating stroke. Nevertheless, the metabolite signatures and underlying mechanisms associated with EDA remain unclear, which deserve further elucidation for improving the accurate usage of EDA. Ischemia stroke was simulated by intraluminal occlusion of the right middle cerebral artery for 1 h, followed by reperfusion for 24 h in mice. Brain infarct size, neurological deficits, and lactate dehydrogenase (LDH) levels were improved by EDA. Significantly differential metabolites were screened with untargeted metabolomics by cross-comparisons with pre- and posttreatment of EDA under cerebral ischemia/reperfusion (I/R) injury. The possibly involved pathways, such as valine, leucine, and isoleucine biosynthesis, and phenylalanine, taurine, and hypotaurine metabolisms, were enriched with differential metabolites and relevant regulatory enzymes, respectively. The network of differential metabolites was constructed for the integral exhibition of metabolic characteristics. Targeted analysis of taurine, an important metabolic marker, was performed for further validation. The level of taurine decreased in the MCAO/R group and increased in the EDA group. The inhibition of EDA on cerebral endothelial cell apoptosis was confirmed by TdT-mediated dUTP nick-end labeling (TUNEL) stain. Cysteine sulfinic acid decarboxylase (CSAD), the rate-limiting enzyme of taurine generation, significantly increased along with inhibiting endothelial cell apoptosis after treatment of EDA. Thus, CSAD, as the possible new therapeutic target of EDA, was selected and validated by Western blot and immunofluorescence. Together, this study provided the metabolite signatures and identified CSAD as an unrecognized therapeutic intervention for EDA in the treatment of ischemic stroke via inhibiting brain endothelial cell apoptosis.

Systematic Screening for SARS-CoV-2 to Detect Asymptomatic Infections: An Epitome of Taiwan's Outbreak.

BACKGROUND: Increased studies have revealed that asymptomatic carriers substantially impact the epidemic and that asymptomatic transmission is very common. Therefore, the asymptomatic transmission threat to the spread of the pandemic should not be neglected. METHODS: The local outbreak in Taiwan, especially in Taipei City, is unprecedented and paramount and has claimed hundreds of lives, tens of thousands of cases, and enormous economic costs. As care providers and gatekeepers of infectious diseases, Taipei City Hospital has to perform regular polymerase chain reaction (PCR) results of admitted patients and healthcare workers (HCWs) to achieve these goals. RESULTS: In this study, the results revealed a low positive rate of less than 1%, but the asymptomatic proportions could range from 42% to 46%, which bolsters that systematic screening was effective in controlling coronavirus disease-19 (COVID-19) of Novel Coronavirus or Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) and might be an exemplar to other similar scenarios. Universal screening of admitted patients may be important and necessary, especially in asymptomatic patients. CONCLUSIONS: Regular screening for healthcare providers is also important during this pandemic, and it is recommended that admitted patients and healthcare providers undergo systemic PCR testing.

Structural characterization of a water-soluble polysaccharide from Angelica dahurica and its antitumor activity in H22 tumor-bearing mice.

A novel Angelica dahurica polysaccharide (ADP) with Mw of 6.09 × 103 Da was isolated. The contents of total sugar and uronic acid in ADP were 91.04% and 12.69%. The structure characteristics indicated that ADP was an acidic polysaccharide consisting of rhamnose, arabinose, galactose, glucose, mannose, glucuronic acid and galacturonic acid (0.09: 0.61: 1.88: 1: 0.14: 0.63: 0.03). Moreover, there were →3)-Manp-(1→, →4, 6)-Galp-(1→, →4)-Galp-(1→, →3)-Glcp-(1→, →5)-Araf-(1→, →2)-Galp-(1→ in ADP with relative molar ratios of 0.32:0.57:0.29:0.95:0.71:0.26. In vivo experiments suggested that ADP significantly inhibited the tumor growth of mice, increased the activities of spleen lymphocytes and natural killer (NK) cells, improved the cytokine level (IL-2 and TNF-α) and the proportions of lymphocyte subsets in the peripheral blood. The tumor cell progression was arrested in the G1 phase, and the apoptosis rate of tumor cells were 7.54% and 19.32% at the dose of 100 and 200 mg/kg, which was consistent with the results of pathological observation. In summary, the study might provide a theoretical basis for the application on functional foods containing Angelica dahurica polysaccharides.

A novel polysaccharide from Castanea mollissima Blume: Preparation, characteristics and antitumor activities in vitro and in vivo.

A new water-soluble polysaccharide, CMP90, with a molecular weight of 23.9 kDa was isolated from Castanea mollissima Blume and the preliminary structural characteristics and antitumor effects of CMP90 in vitro and in vivo were investigated in the research. CMP90 consists of arabinose, galactose, glucose, xylose and mannose (molar ratio: 0.08:0.11:5.14:0.12:0.08) with α- and ß-anomeric units. The results of in vitro experiments indicated that CMP90 exhibited a significant inhibitory effect on the proliferation of HL-60 cells with typical apoptotic characteristics by inducing cell cycle arrested at G1/M phase. Additionally, the results in vivo suggested CMP90 was able to inhibit the growth of S180 solid tumors via protecting immune organs, improving the levels of serum cytokines (TNF-α, IL-2 and IFN-γ), enhancing the activities of immune cells (macrophages, lymphocytes and NK cells) and inducing cell apoptosis or death. Taken together, these combined data clearly indicated that CMP90 may be used as a potential candidate agent for cancer therapy.

Kanglexin, a novel anthraquinone compound, protects against myocardial ischemic injury in mice by suppressing NLRP3 and pyroptosis.

Pyroptosis is a form of inflammatory cell death that could be driven by the nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation following myocardial infarction (MI). Emerging evidence suggests the therapeutic potential for ameliorating MI-induced myocardial damages by targeting NLRP3 and pyroptosis. In this study, we investigated the myocardial protection effect of a novel anthraquinone compound (4,5-dihydroxy-7-methyl-9,10-anthraquinone-2-ethyl succinate) named Kanglexin (KLX) in vivo and in vitro. Male C57BL/6 mice were pre-treated either with KLX (20, 40 mg· kg-1per day, intragastric gavage) or vehicle for 7 consecutive days prior to ligation of coronary artery to induce permanent MI. KLX administration dose-dependently reduced myocardial infarct size and lactate dehydrogenase release and improved cardiac function as compared to vehicle-treated mice 24 h after MI. We found that MI triggered NLRP3 inflammasome activation leading to conversion of interleukin-1ß (IL-1ß) and IL-18 into their active mature forms in the heart, which could expand the infarct size and drive cardiac dysfunction. We also showed that MI induced pyroptosis, as evidenced by increased DNA fragmentation, mitochondrial swelling, and cell membrane rupture, as well as increased levels of pyroptosis-related proteins, including gasdermin D, N-terminal GSDMD, and cleaved caspase-1. All these detrimental alterations were prevented by KLX. In hypoxia- or lipopolysaccharide (LPS)-treated neonatal mouse ventricular cardiomyocytes, we showed that KLX (10 µM) decreased the elevated levels of terminal deoxynucleotidyl transferase dUTP nick end labeling- and propidium iodide-positive cells, and pyroptosis-related proteins. We conclude that KLX prevents MI-induced cardiac damages and cardiac dysfunction at least partly through attenuating NLRP3 and subsequent cardiomyocyte pyroptosis, and it is worthy of more rigorous investigations for its potential for alleviating ischemic heart disease.

Rapid discovery and identification of the anti-inflammatory constituents in Zhi-Shi-Zhi-Zi-Chi-Tang.

The anti-inflammatory active ingredients of Zhi-Shi-Zhi-Zi-Chi-Tang (ZSZZCT), a traditional Chinese medicine formula, were predicted and identified using an approach based on activity index, LC-MS, semi-preparative LC and NMR. Firstly, the whole extract of ZSZZCT was analyzed using liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) and liquid chromatography - ion trap mass spectrometry (LC-IT-MS), 79 constituents were detected and 39 constituents were identified unambiguously or tentatively. Subsequently, the whole extract of the formula was separated into multiple components and the activity index method was used to calculate index values of the 79 constituents by integrating the chemical and pharmacological information of multiple components. Four polymethoxyl flavones were predicted as the major active constituents according to the activity index values. Furthermore, three polymethoxyl flavones were prepared using the strategy with semi-preparative LC guided by LC-MS, and their anti-inflammatory activities were validated. The results show that three polymethoxyl flavones with higher positive index values, i.e., 3, 5, 6, 7, 8, 3', 4'-heptamethoxyflavone, 3-hydroxynobiletein and tangeretin had significant anti-inflammatory effects. In conclusion, the predicted results indicated that the activity index method is feasible for the accurate prediction of active constituents in TCM formulae.

Downregulated serum miR-223 servers as biomarker in Alzheimer's disease.

Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is characterized by progressive memory loss and deteriorated higher cognitive functions. An economical, rapid and noninvasive biomarker for AD has not been identified. We aimed to investigate the diagnostic value of serum miR-223 and miR-519 in AD. The expressions of miR-223 and miR-519, with previously reported AD-associated miR-29 and miR-125b, were measured by quantitative reverse transcription polymerase chain reaction in the serum of 84 probable sporadic AD patients (age onset > 65 years) and 62 healthy control populations in China. Analyses were undertaken to assess the specificity and sensitivity of miRNAs to predict AD. In addition, the relationship between miRNAs and mini mental state examination (MMSE) scores in AD patients was also assessed. Serum miR-29, miR-125b and miR-223 were significantly decreased, but serum miR-519 was significantly increased in AD patients compared with healthy blood donors. In addition, serum miR-223 was strongly positively correlated with MMSE score in AD patients but serum miR-519 was not. Importantly, the receiver operating characteristic (ROC) result of serum miR-223 for prediction of AD was 0.786, higher than those of serum miR-29 (0.734) or miR-125b (0.726). The combination of serum miR-223 and miR-125b gave improved sensitivity/specificity for AD prediction (area under the ROC curve, 0.879) than either miRNA alone. Our preliminary findings indicate that serum miR-223 might be a potential biomarker for AD evaluation. Copyright © 2016 John Wiley & Sons, Ltd.

Comparison of the analgesic effects of cryoanalgesia vs. parecoxib for lung cancer patients after lobectomy.

This study was designed to compare the analgesic effects of cryoanalgesia and parecoxib in lung cancer patients after lobectomy. A total of 178 lung cancer patients awaiting large-sized lobectomy were enrolled in the study. The patients were randomly divided into Group A (intercostal nerve cryoanalgesia) and Group B (parecoxib). The analgesic and adverse effects were compared between the two groups. The pain score of Group A was significantly lower than that of Group B (P < 0.05). The patients in Group A used significantly less morphine than those in Group B (P < 0.05). There were also significantly fewer complications in Group A than in Group B (P < 0.05). Cryoanalgesia of the intercostal nerves can be considered an economical, safe and simple technique for the long-term management of post-lobectomy pain.

Characterization of in vitro modified human very low-density lipoprotein particles and phospholipids by capillary electrophoresis.

A simple capillary zone electrophoresis (CZE) method was used to characterize human very low-density lipoprotein (VLDL) particles for four healthy donors. One major peak was observed for native, in vitro oxidized and glycated VLDL particles. The effective mobilities and peak areas of the capillary electrophoresis (CE) profiles showed good reproducibility and precision. The mobility of the oxidized VLDL peak was higher than that of the native VLDL. The mobility of the glycated VLDL peak was similar to that of the native VLDL. Phospholipids isolated from VLDL particles were analyzed by our recently developed micellar electrokinetic chromatography (MEKC) with a high-salt stacking method. At absorbance 200 nm, the native VLDL phospholipids showed a major peak and a minor peak for each donor. For oxidized VLDL phospholipids, the area of the major peak reduced for three donors, possibly due to phospholipid decomposition. For glycated VLDL phospholipids, the peak mobilities were more positive than native VLDL phospholipids for two donors, possibly due to phospholipid-linked advanced glycation end products (AGEs). Very interestingly, at absorbance 234 nm, the major peak of oxidized VLDL phospholipids was resolved as two peaks for each donor, possibly due to conjugated dienes formed upon oxidation.

The body weight loss during acute exposure to high-altitude hypoxia in sea level residents.

Weight loss is frequently observed after acute exposure to high altitude. However, the magnitude and rate of weight loss during acute exposure to high altitude has not been clarified in a controlled prospective study. The present study was performed to evaluate weight loss at high altitude. A group of 120 male subjects [aged (32±6) years] who worked on the construction of the Golmud-Lhasa Railway at Kunlun Mountain (altitude of 4 678 m) served as volunteer subjects for this study. Eighty-five workers normally resided at sea level (sea level group) and 35 normally resided at an altitude of 2 200 m (moderate altitude group). Body weight, body mass index (BMI), and waist circumference were measured in all subjects after a 7-day stay at Golmud (altitude of 2 800 m, baseline measurements). Measurements were repeated after 33-day working on Kunlun Mountain. In order to examine the daily rate of weight loss at high altitude, body weight was measured in 20 subjects from the sea level group (sea level subset group) each morning before breakfast for 33 d at Kunlun Mountain. According to guidelines established by the Lake Louise acute mountain sickness (AMS) consensus report, each subject completed an AMS self-report questionnaire two days after arriving at Kunlun Mountain. After 33-day stay at an altitude of 4 678 m, the average weight loss for the sea level group was 10.4% (range 6.5% to 29%), while the average for the moderate altitude group was 2.2% (-2% to 9.1%). The degree of weight loss (Δ weight loss) after a 33-day stay at an altitude of 4 678 m was significantly correlated with baseline body weight in the sea level group (r=0.677, P<0.01), while the correlation was absent in the moderate altitude group (r=0.296, P>0.05). In the sea level subset group, a significant weight loss was observed within 20 d, but the weight remained stable thereafter. AMS-score at high altitude was significantly higher in the sea level group (4.69±2.48) than that in the moderate altitude group (2.97±1.38), and was significantly correlated with baseline body weight. These results indicate that (1) the person with higher body weight during stay at high altitude loses more weight, and this is more pronounced in sea level natives when compared with that in moderate altitude natives; (2) heavier individuals are more likely to develop AMS than leaner individuals during exposure to high-altitude hypoxia.

[Application of "overlay-maps" model in prior period research of ecological planning of Shanghai Expo area].

An ecological planning approach for Shanghai Expo 2010 was constructed based on "overlay-maps" model, and the ecological elements and GIS visualization in Shanghai Expo 2010 area were investigated and analyzed from the aspects of human comfort degree, life health, and sustainable utilization of resources. This approach included the determination of objectives for ecological planning, the selection and in situ investigation of ecological elements, the construction of eco-database, and the integrative analysis of GIS visualization, being indispensable for the prior period research of ecological planning of Expo area. Based on the present situation of soil pollution in the Expo area, a tentative scheme of soil restoration and utilization was brought forward, with the concerns of green space demand and soil secondary pollution avoidance. To protect the wild life habitats in Expo area, a demarcation of conservation areas for plants and original landscape was made. A conception of using landscape elements to optimize air temperature, humidity, and ventilation in the process of urban design was proposed, aimed to promote the human comfort degree under tropical monsoon conditions.

[Microclimate and comfortable degree of Shanghai urban open spaces in summer].

Based on the observation data of air temperature, relative humidity, wind speed, and solar radiation from May to August 2006, the regulation effects of five types of open spaces (square, fountain, grassplot, corridor, and woodland) in Shanghai urban districts on the microclimate were analyzed, and discomfort index (DI) was introduced to evaluate the effects of these five types of open spaces on human body' s comfortable degree. The results showed that there existed definite differences in the air temperature and relative humidity among the open spaces, with the mean temperature decreased in the order of square > grassplot > fountain > corridor > woodland, and the mean relative humidity decreased in the order of woodland > corridor > fountain > grassplot > square. The area of the square, the wind speed and direction near the fountain, the grass species on the grass-plot, the width and tree coverage of the corridor, and the tree coverage and canopy height of the woodland had significant correlations with the microclimate parameters of corresponding open spaces. Comparing with other three types of open spaces, woodland and corridor had better regulation effects on the microclimate via shading, decreasing air temperature, and increasing relative humidity.