Assessing the impact of circulating inflammatory cytokines and proteins as drivers and therapeutic targets in epilepsy: A Mendelian randomization study.

BACKGROUND: Previous research has demonstrated that neuroinflammation is a key element in the progress of epilepsy. Nevertheless, it is currently unidentified which inflammatory factors and proteins increase or decrease the risk of epilepsy. METHODS: We adopted Mendelian randomization techniques to explore the causal relationship between circulating inflammatory factors and proteins and various epilepsy. Our principal approach was inverse variance weighting, supplemented by several sensitivity analyses to guarantee the robustness of our findings. RESULTS: Studies have identified associations between epilepsy and specific inflammatory factors and proteins: three inflammatory factors and six proteins are linked to epilepsy in general; one inflammatory factor and four proteins are associated with focal epilepsy with no documented lesions; two inflammatory factors and three proteins are related to focal epilepsy, excluding cases with hippocampal sclerosis; two inflammatory factors and two proteins are connected to juvenile myoclonic epilepsy; two inflammatory factors and five proteins are linked to juvenile absence epilepsy; four inflammatory proteins are associated with childhood absence epilepsy; two inflammatory factors are related to focal epilepsy overall; two inflammatory factors and two proteins are connected to generalized epilepsy; and two inflammatory proteins are linked to generalized epilepsy with tonic-clonic seizures. Additionally, six inflammatory factors may play a downstream role in focal epilepsy. CONCLUSION: Our study uncovers various inflammatory factors and proteins that influence the risk of epilepsy, offering instructive insights to the diagnosis and therapy of the condition.

Application of LRG mechanism in normal pressure hydrocephalus.

Normal pressure hydrocephalus (NPH) is a prevalent type of hydrocephalus, including secondary normal pressure hydrocephalus (SNPH) and idiopathic normal pressure hydrocephalus (INPH). However, its clinical diagnosis and pathological mechanism are still unclear. Leucine-rich α-2 glycoprotein (LRG) is involved in various human diseases, including cancer, diabetes, cardiovascular disease, and nervous system diseases. Now the physiological mechanism of LRG is still being explored. According to the current research results on LRG, we found that the agency of LRG has much to do with the known pathological process of NPH. This review focuses on analyzing the LRG signaling pathways and the pathological mechanism of NPH. According to the collected literature evidence, we speculated that LRG probably be involved in the pathological process of NPH. Finally, based on the mechanism of LRG and NPH, we also summarized the evidence of molecular targeted therapies for future research and clinical application.

IMH-Net: a convolutional neural network for end-to-end EEG motor imagery classification.

As the main component of Brain-computer interface (BCI) technology, the classification algorithm based on EEG has developed rapidly. The previous algorithms were often based on subject-dependent settings, resulting in BCI needing to be calibrated for new users. In this work, we propose IMH-Net, an end-to-end subject-independent model. The model first uses Inception blocks extracts the frequency domain features of the data, then further compresses the feature vectors to extract the spatial domain features, and finally learns the global information and classification through Multi-Head Attention mechanism. On the OpenBMI dataset, IMH-Net obtained 73.90 ± 13.10% accuracy and 73.09 ± 14.99% F1-score in subject-independent manner, which improved the accuracy by 1.96% compared with the comparison model. On the BCI competition IV dataset 2a, this model also achieved the highest accuracy and F1-score in subject-dependent manner. The IMH-Net model we proposed can improve the accuracy of subject-independent Motor Imagery (MI), and the robustness of the algorithm is high, which has strong practical value in the field of BCI.

Research progress on the role of the Wnt signaling pathway in pituitary adenoma.

Pituitary adenoma (PA) is the third most common central nervous system tumor originating from the anterior pituitary, but its pathogenesis remains unclear. The Wnt signaling pathway is a conserved pathway involved in cell proliferation, Self-renewal of stem cells, and cell differentiation. It is related to the occurrence of various tumors, including PA. This article reviews the latest developments in Wnt pathway inhibitors and pathway-targeted drugs. It discusses the possibility of combining Wnt pathway inhibitors with immunotherapy to provide a theoretical basis for the combined treatment of PA.

Anastasis enhances metastasis and chemoresistance of colorectal cancer cells through upregulating cIAP2/NFκB signaling.

Chemotherapy is a common strategy to treat cancer. However, acquired resistance and metastasis are the major obstacles to successful treatment. Anastasis is a process by which cells survive executioner caspase activation when facing apoptotic stress. Here we demonstrate that colorectal cancer cells can undergo anastasis after transient exposure to chemotherapeutic drugs. Using a lineage tracing system to label and isolate cells that have experienced executioner caspase activation in response to drug treatment, we show that anastasis grants colorectal cancer cells enhanced migration, metastasis, and chemoresistance. Mechanistically, treatment with chemotherapeutic drugs induces upregulated expression of cIAP2 and activation of NFκB, which are required for cells to survive executioner caspase activation. The elevated cIAP2/NFκB signaling persists in anastatic cancer cells to promote migration and chemoresistance. Our study unveils that cIAP2/NFκB-dependent anastasis promotes acquired resistance and metastasis after chemotherapy.

Chemotherapy-induced executioner caspase activation increases breast cancer malignancy through epigenetic de-repression of CDH12.

Cancer relapse and metastasis are major obstacles for effective treatment. One important mechanism to eliminate cancer cells is to induce apoptosis. Activation of executioner caspases is the key step in apoptosis and was considered "a point of no return". However, in recent years, accumulating evidence has demonstrated that cells can survive executioner caspase activation in response to apoptotic stimuli through a process named anastasis. Here we show that breast cancer cells that have survived through anastasis (anastatic cells) after exposure to chemotherapeutic drugs acquire enhanced proliferation and migration. Mechanistically, cadherin 12 (CDH12) is persistently upregulated in anastatic cells and promotes breast cancer malignancy via activation of ERK and CREB. Moreover, we demonstrate that executioner caspase activation induced by chemotherapeutic drugs results in loss of DNA methylation and repressive histone modifications in the CDH12 promoter region, leading to increased CDH12 expression. Our work unveils the mechanism underlying anastasis-induced enhancement in breast cancer malignancy, offering new therapeutic targets for preventing post-chemotherapy cancer relapse and metastasis.

The Study of Overexpression of Peroxiredoxin-2 Reduces MPP+-Induced Toxicity in the Cell Model of Parkinson's Disease.

Parkinson's disease (PD) is a chronic neurodegenerative disorder characterized by dopaminergic neuron loss, which is related to excessive reactive oxygen species (ROS) accumulation. Endogenous peroxiredoxin-2 (Prdx-2) has potent anti-oxidative and anti-apoptotic effects. Proteomics studies revealed plasma levels of Prdx-2 were significantly lower in PD patients than in healthy individuals. For further study of the activation of Prdx-2 and its role in vitro, SH-SY5Y cells and the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) were used to model PD. ROS content, mitochondrial membrane potential, and cell viability were used to assess the effect of MPP+ in SH-SY5Y cells. JC-1 staining was used to determine mitochondrial membrane potential. ROS content was detected using a DCFH-DA kit. Cell viability was measured using the Cell Counting Kit-8 assay. Western blot detected the protein levels of tyrosine hydroxylase (TH), Prdx-2, silent information regulator of transcription 1 (SIRT1), Bax, and Bcl-2. The results showed that MPP+-induced accumulation of ROS, depolarization of mitochondrial membrane potential, and reduction of cell viability occurred in SH-SY5Y cells. In addition, the levels of TH, Prdx-2, and SIRT1 decreased, while the ratios of Bax and Bcl-2 increased. Then, Prdx-2 overexpression in SH-SY5Y cells showed significant protection against MPP+ -induced neuronal toxicity, as evidenced by the decrease in ROS content, increase in cell viability, increase in the level of TH, and decrease in the ratios of Bax and Bcl-2. Meanwhile, SIRT1 levels increase with the level of Prdx-2. This suggests that the protection of Prdx-2 may be related to SIRT1. In conclusion, this study indicated that overexpression of Prdx-2 reduces MPP+-induced toxicity in SH-SY5Y cells and may be mediated by SIRT1.

Molecular chaperone GroEL-GroES enhances the soluble expression of biologically active ovine growth hormone in the prokaryotic system.

Growth hormone (GH) plays important roles in growth and development of mammalian animals and is valuable for many applications. This study aimed to express and purify biological active recombinant ovine growth hormone (roGH) through prokaryotic expression system. The roGH coding sequence was ligated into the prokaryotic expression vector and transformed into Escherichia coli (E. coli) for protein expression. Factors that influence the roGH expression were examined and the appropriate culture temperature (20 °C) and inducer (IPTG) concentration (25 µM) were determined. To enhance the soluble expression of the protein, co-expression with the molecular chaperone GroEL-GroES was utilized and eventually achieved a high yield of soluble roGH expressed in E. coli. Further, the fusion tag in expressed target protein could be efficiently removed through thrombin-specific cleavage. The expressed roGH was identified by Western blotting and the LC-MS spectrum confirmed its molecular weight of 22749.22 Da. Finally, the purified roGH had an expected biological activity when assayed in cell models in vitro and experimental mouse in vivo. In conclusion, the present study established an efficient and simple approach to produce recombinant GH, and facilitate relevant research and applications.

Polyphenylene Sulfide-Based Solid-State Separator for Limited Li Metal Battery.

The urgent need for high energy batteries is pushing the battery studies toward the Li metal and solid-state direction, and the most central question is finding proper solid-state electrolyte (SSE). So far, the recently studied electrolytes have obvious advantages and fatal weaknesses, resulting in indecisive plans for industrial production. In this work, a thin and dense lithiated polyphenylene sulfide-based solid state separator (PPS-SSS) prepared by a solvent-free process in pilot stage is proposed. Moreover, the PPS surface is functionalized to immobilize the anions, increasing the Li+ transference number to 0.8-0.9, and widening the electrochemical potential window (EPW > 5.1 V). At 25 °C, the PPS-SSS exhibits high intrinsic Li+ diffusion coefficient and ionic conductivity (>10-4 S cm-1 ), and Li+ transport rectifying effect, resulting in homogenous Li-plating on Cu at 2 mA cm-2 density. Based on the limited Li-plated Cu anode or anode-free Cu, high loadings cathode and high voltage, the Li-metal batteries (LMBs) with polyethylene (PE) protected PPS-SSSs deliver high energy and power densities (>1000 Wh L-1 and 900 W L-1 ) with >200 cycling life and high safety, exceeding those of state-of-the-art Li-ion batteries. The results promote the Li metal battery toward practicality.

[Removal Efficiency and Mechanism of Ammonia Nitrogen in a Low Temperature Groundwater Purification Process].

To explore the mechanism and efficiency of ammonia nitrogen removal, a pilot-scale biofilter for the simultaneous removal of high concentrations of iron, manganese, and ammonia nitrogen[Fe(Ⅱ) 11.9-14.8 mg·L-1, Mn(Ⅱ) 1.1-1.5mg·L-1, and NH4+-N 1.1-3.2 mg·L-1] from low temperature(5-6℃) groundwater was operated in a water supply plant in Northeast China. Results indicated excellent performance for ammonia nitrogen removal during the initial start-up stage. According to theoretical analysis and experimental verification, TNloss was driven by the adsorption of ammonia nitrogen by iron oxides, and the conversion of ammonia nitrogen into nitrate nitrogen occurred via biological nitrification. When the concentration of ammonia nitrogen increased, due to limited adsorption sites, the adsorption capacity of iron oxides remained stable at approximately 1 mg·L-1. For the same period, the amount of ammonia nitrogen removal via oxidation continued to increase, with higher quantities removed in the upper filter layer than in the lower filter layer. Dissolved oxygen(DO) is the limiting factor in the further increase in the removal of ammonia nitrogen by oxidation. With an increase in the filtration rate, the adsorption time of ammonia nitrogen by iron oxides was shortened, and the adsorption amount was reduced. Meanwhile, the shortening of EBCT reduced the ammonia nitrogen removed by nitrification under the action of nitrifying bacteria in the unit volume of the filter material. Based on these findings, it is recommended that the thickness of the filter layer should be increased to improve ammonia nitrogen removal performance.

DNA repair glycosylase hNEIL1 triages damaged bases via competing interaction modes.

DNA glycosylases must distinguish the sparse damaged sites from the vast expanse of normal DNA bases. However, our understanding of the nature of nucleobase interrogation is still limited. Here, we show that hNEIL1 (human endonuclease VIII-like 1) captures base lesions via two competing states of interaction: an activated state that commits catalysis and base excision repair, and a quarantine state that temporarily separates and protects the flipped base via auto-inhibition. The relative dominance of the two states depends on key residues of hNEIL1 and chemical properties (e.g. aromaticity and hydrophilicity) of flipped bases. Such a DNA repair mechanism allows hNEIL1 to recognize a broad spectrum of DNA damage while keeps potential gratuitous repair in check. We further reveal the molecular basis of hNEIL1 activity regulation mediated by post-transcriptional modifications and provide an example of how exquisite structural dynamics serves for orchestrated enzyme functions.

Detect-seq reveals out-of-protospacer editing and target-strand editing by cytosine base editors.

Cytosine base editors (CBEs) have the potential to correct human pathogenic point mutations. However, their genome-wide specificity remains poorly understood. Here we report Detect-seq for the evaluation of CBE specificity. It enables sensitive detection of CBE-induced off-target sites at the genome-wide level. Detect-seq leverages chemical labeling and biotin pulldown to trace the editing intermediate deoxyuridine, thereby revealing the editome of CBE. In addition to Cas9-independent and typical Cas9-dependent off-target sites, we discovered edits outside the protospacer sequence (that is, out-of-protospacer) and on the target strand (which pairs with the single-guide RNA). Such unexpected off-target edits are prevalent and can exhibit a high editing ratio, while their occurrences exhibit cell-type dependency and cannot be predicted based on the sgRNA sequence. Moreover, we found out-of-protospacer and target-strand edits nearby the on-target sites tested, challenging the general knowledge that CBEs do not induce proximal off-target mutations. Collectively, our approaches allow unbiased analysis of the CBE editome and provide a widely applicable tool for specificity evaluation of various emerging genome editing tools.

A simple and rapid diagnostic method for 13 types of high-risk human papillomavirus (HR-HPV) detection using CRISPR-Cas12a technology.

Cervical carcinoma is the second most common cancer in women worldwide with greater than 99% of the cases caused by human papillomaviruses (HPVs). Early detection of HPVs especially the high risk types (HR-HPVs) are essential to prevent the disease progression. The existing methods for HPV detection, such as qPCR are of high sensitivity and specificity, but the need for expensive machinery and well-trained personnel slow down the disease detection. The emerging Cas12a-based method presents a new technique for nucleic acid detection. However, it is time-consuming and labor-intensive when used for HPV detection, as several reactions are required in order to identify multiple HPV infections. We herein present a non-genotyping method for 13 types of HR-HPV detection in a single reaction by combining the isothermal recombinase polymerase amplification (RPA) method with CRISPR-Cas12a technology. The result could be achieved in 35 min with high sensitivity (500 copies per reaction). This assay represents great advances for the application of RPA-Cas12a system and holds a great potential to address the key challenges facing the HPV diagnostics.

An enhanced method for nucleic acid detection with CRISPR-Cas12a using phosphorothioate modified primers and optimized gold-nanopaticle strip.

CRISPR-Cas12a system has been shown promising for nucleic acid diagnostics due to its rapid, portable and accurate features. However, cleavage of the amplicons and primers by the cis- and trans-activity of Cas12a hinders the attempts to integrate the amplification and detection into a single reaction. Through phosphorothioate modification of primers, we realized onepot detection with high sensitivity using plasmids of SARS-CoV-2, HPV16 and HPV18. We also identified the activated Cas12a has a much higher affinity to C nucleotide-rich reporter than others. By applying such reporters, the reaction time required for a lateral-flow readout was significantly reduced. Furthermore, to improve the specificity of the strip-based assay, we created a novel reporter and, when combined with a customized gold-nanopaticle strip, the readout was greatly enhanced owing to the elimination of the nonspecific signal. This established system, termed Targeting DNA by Cas12a-based Eye Sight Testing in an Onepot Reaction (TESTOR), was validated using clinical cervical scrape samples for human papillomaviruses (HPVs) detection. Our system represents a general approach to integrating the nucleic acid amplification and detection into a single reaction in CRISPR-Cas systems, highlighting its potential as a rapid, portable and accurate detection platform of nucleic acids.

Microbial dynamics, metabolomic profiles, and the correlation between them during fermentation of serofluid dish.

BACKGROUND: Serofluid dish, a traditional Chinese fermented food, possesses unique flavors and health beneficial effects. These properties are likely due to the sophisticated metabolic networks during fermentation, which are mainly driven by microbiota. However, the exact roles of metabolic pathways and the microbial community during this process remain equivocal. RESULTS: Here, we investigated the microbial dynamics by next-generation sequencing, and outlined a differential non-targeted metabolite profiling in the process of serofluid dish fermentation using the method of hydrophilic interaction liquid chromatography column with ultra-high-performance liquid chromatography-quadruple time-of-flight mass spectrometry. Lactobacillus was the leading genus of bacteria, while Pichia and Issatchenkia were the dominant fungi. They all accumulated during fermentation. In total, 218 differential metabolites were identified, of which organic acids, amino acids, sugar and sugar alcohols, fatty acids, and esters comprised the majority. The constructed metabolic network showed that tricarboxylic acid cycle, urea cycle, sugar metabolism, amino acids metabolism, choline metabolism, and flavonoid metabolism were regulated by the fermentation. Furthermore, correlation analysis revealed that the leading fungi, Pichia and Issatchenkia, were linked to organic acids, amino acid and sugar metabolism, flavonoids, and several other flavor and functional components. Antibacterial tests indicated the antibacterial effect of serofluid soup against Salmonella and Staphylococcus. CONCLUSION: This work provides new insights into the complex microbial and metabolic networks during serofluid dish fermentation, and a theoretical basis for the optimization of its industrial production. © 2020 Society of Chemical Industry.

[Effect of Filter Speed and Water Quality on Ammonia Removal in Groundwater Containing Iron, Manganese, and Ammonia at Low Temperature].

In a groundwater plant we carried out a process operation test of biological removal of iron and manganese nitrification coupled with completely autotrophic ammonium removal over nitrite (CANON) (Fe(Ⅱ) 2.91-6.35 mg·L-1, Mn(Ⅱ) 0.47-0.98 mg·L-1, NH4+-N 1.15-2.26 mg·L-1) at low temperature (6-8℃), to explore the effects of filter speed and water quality on ammonia nitrogen removal. The results showed that the mature low-temperature biological filter column, which had been out of service for one month, was cultured for 40 days at a filtration rate of 2 m·h-1 and successfully started. In this process, when the water inlet concentration remained the same, the improved filter speed would reduce the efficiency of ammonia nitrogen capture by the filter column, increase the concentration of ammonia nitrogen in the depth of the filter layer, and improve the efficiency of ammonia nitrogen ions capture by anaerobic ammonia oxidation bacteria (AnAOB) in the depth of the filter layer, so that the ammonia nitrogen removed by CANON in the water increased, while the ammonia nitrogen removed by nitrification decreased. When the filter speed remained unchanged, the concentration of ammonia nitrogen in water was increased to make the ammonia nitrogen with higher concentration enter the filter layer, which increased the concentration of ammonia nitrogen in the zone where ammonia nitrogen and nitrous nitrogen coexist, and improved the net catching efficiency of AnAOB on ammonia nitrogen ions in the filter layer, thus resulting in an increase in ammonia nitrogen removed by CANON.

[Low Temperature Ammonia Nitrogen Removal from an Iron, Manganese, and Ammonia Groundwater Purification Process with Different Concentrations of Iron and Manganese].

In a groundwater plant, removal of iron, manganese, and ammonia nitrogen was performed via a purification process using a filter column at a low temperature (5-6℃). Iron, manganese and ammonia [Fe(Ⅱ) 0-19.26 mg·L-1, Mn(Ⅱ) 0.52-2.05 mg·L-1, and NH4+-N 0.37-2.59 mg·L-1] were analyzed to explore the ammonia nitrogen removal efficiency under different iron and manganese concentrations. The results showed that when the concentration of manganese in the inlet water was maintained at approximately 0.6 mg·L-1 and the concentration of ferrous iron in the inlet water was increased, with the increase of iron oxides in the filter layer, the ratio of ammonia nitrogen removed by adsorption of iron oxides increased, while the ratio of ammonia nitrogen removed by nitrification will decreased and adsorption preceded nitrification. When the concentration of ferrous iron in the water was maintained at approximately 8 mg·L-1and 11 mg·L-1, and the concentration of manganese in the water was increased, the proportion of ammonia nitrogen removed by adsorption did not increase with the increase of manganese oxide, and the removal route of ammonia nitrogen hardly changed. This is because less manganese oxides were formed 20 cm before the filter layer, which had little effect on the ammonia nitrogen adsorbed in this range. The production area of manganese oxides was concentrated below 20 cm of the filter layer, and most ammonia nitrogen was removed by adsorption and nitrification before this area, and the manganese oxides in this area did not adsorb ammonia nitrogen.

Resonant broadband unidirectional light scattering based on genetic algorithm.

The spectrum overlapping of the radiative power between magnetic and electric dipole moments in nanoparticles can be used to realize unidirectional light scattering, which is promising for various kinds of applications. Nevertheless, it is still challenging to achieve such overlapping in a broadband manner. Herein, we propose that the combination of a genetic algorithm, Maxwell's equations, and electromagnetic multipole expansion can be used to design a nanoparticle that supports resonant broadband forward light scattering. Microwave experiments are performed to demonstrate our numerical results. The proposed method is quite general, and it can be straightforwardly generalized to design functional unidirectional scatters.

Structural Insights into the Specific Recognition of 5-methylcytosine and 5-hydroxymethylcytosine by TAL Effectors.

Transcription activator-like effectors (TALEs) recognize DNA through repeat-variable diresidues (RVDs), and TALE-DNA interactions are sensitive to DNA modifications. Our previous study deciphered the recognition of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) by TALEs. Here, we report seven crystal structures of TALE-DNA complexes. The 5mC-specific RVD HA recognizes 5mC through van der Waals interactions and exhibits highly similar loop conformation to natural RVDs. The degenerate RVD RG contacts 5mC and 5hmC via van der Waals interactions as well; however, its loop conformation differs significantly. The loop conformations of universal RVD R* and 5hmC-specific RVD Q* are similar to that of RG, while the interactions of R* with C/5mC/5hmC and Q* with 5hmC are mediated by waters. Together, our findings illustrate the molecular basis for the specific recognition of 5mC and 5hmC by multiple noncanonical TALEs and provide insights into the plasticity of the TALE RVD loops.

Great Enhancement of Carbon Energy Storage through Narrow Pores and Hydrogen-Containing Functional Groups for Aqueous Zn-Ion Hybrid Supercapacitor.

The proton transfer mechanism on the carbon cathode surface has been considered as an effective way to boost the electrochemical performance of Zn-ion hybrid supercapacitors (SCs) with both ionic liquid and organic electrolytes. However, cheaper, potentially safer, and more environmental friendly supercapacitor can be achieved by using aqueous electrolyte. Herein, we introduce the proton transfer mechanism into a Zn-ion hybrid supercapacitor with the ZnSO4 aqueous electrolyte and functionalized activated carbon cathode materials (FACs). We reveal both experimentally and theoretically an enhanced performance by controlling the micropores structure and hydrogen-containing functional groups (-OH and -NH functions) of the activated carbon materials. The Zn-ion SCs with FACs exhibit a high capacitance of 435 F g-1 and good stability with 89% capacity retention over 10,000 cycles. Moreover, the proton transfer effect can be further enhanced by introducing extra hydrogen ions in the electrolyte with low pH value. The highest capacitance of 544 F g-1 is obtained at pH = 3. The proton transfer process tends to take place preferentially on the hydroxyl-groups based on the density functional theory (DFT) calculation. The results would help to develop carbon materials for cheaper and safer Zn-ion hybrid SCs with higher energy.