Inflachromene attenuates seizure severity in mouse epilepsy models via inhibiting HMGB1 translocation.

Epilepsy is not well controlled by current anti-seizure drugs (ASDs). High mobility group box 1 (HMGB1) is a DNA-binding protein in the nucleus regulating transcriptional activity and maintaining chromatin structure and DNA repair. In epileptic brains, HMGB1 is released by activated glia and neurons, interacting with various receptors like Toll-like receptor 4 (TLR4) and downstream glutamatergic NMDA receptor, thus enhancing neural excitability. But there is a lack of small-molecule drugs targeting the HMGB1-related pathways. In this study we evaluated the therapeutic potential of inflachromene (ICM), an HMGB-targeting small-molecule inhibitor, in mouse epilepsy models. Pentylenetetrazol-, kainic acid- and kindling-induced epilepsy models were established in mice. The mice were pre-treated with ICM (3, 10 mg/kg, i.p.). We showed that ICM pretreatment significantly reduced the severity of epileptic seizures in all the three epilepsy models. ICM (10 mg/kg) exerted the most apparent anti-seizure effect in kainic acid-induced epileptic status (SE) model. By immunohistochemical analysis of brain sections from kainic acid-induced SE mice, we found that kainic acid greatly enhanced HMGB1 translocation in the hippocampus, which was attenuated by ICM pretreatment in subregion- and cell type-dependent manners. Notably, in CA1 region, the seizure focus, ICM pretreatment mainly inhibited HMGB1 translocation in microglia. Furthermore, the anti-seizure effect of ICM was related to HMGB1 targeting, as pre-injection of anti-HMGB1 monoclonal antibody (5 mg/kg, i.p.) blocked the seizure-suppressing effect of ICM in kainic acid-induced SE model. In addition, ICM pretreatment significantly alleviated pyramidal neuronal loss and granule cell dispersion in kainic acid-induced SE model. These results demonstrate that ICM is an HMGB-targeting small molecule with anti-seizure potential, which may help develop a potential drug for treating epilepsy.

Cytoplasmic cAMP-sensing domain of hyperpolarization-activated cation (HCN) channels uses two structurally distinct mechanisms to regulate voltage gating.

Voltage gating of hyperpolarization-activated cation (HCN) channels is potentiated by direct binding of cAMP to a cytoplasmic cAMP-sensing domain (CSD). When unliganded, the CSD inhibits hyperpolarization-dependent opening of the HCN channel gate; cAMP binding relieves this autoinhibition so that opening becomes more favorable thermodynamically. This autoinhibition-relief mechanism is conserved with that of several other cyclic nucleotide receptors using the same ligand-binding fold. Besides its thermodynamic effect, cAMP also modulates the depolarization-dependent deactivation rate by kinetically trapping channels in an open state. Here we report studies of strong open-state trapping in an HCN channel showing that the well-established autoinhibition-relief model is insufficient. Whereas deletion of the CSD mimics the thermodynamic open-state stabilization usually associated with cAMP binding, CSD deletion removes rather than mimics the kinetic effect of strong open-state trapping. Substitution of different CSD sequences leads to variation of the degree of open-state trapping in the liganded channel but not in the unliganded channel. CSD-dependent open-state trapping is observed during a voltage-dependent deactivation pathway, specific to the secondary open state that is formed by mode shift after prolonged hyperpolarization activation. This hysteretic activation-deactivation cycle is preserved by CSD substitution, but the change in deactivation kinetics of the liganded channel resulting from CSD substitution is not correlated with the change in autoinhibition properties. Thus the liganded and the unliganded forms of the CSD respectively provide the structural determinants for open-state trapping and autoinhibition, such that two distinct mechanisms for cAMP regulation can operate in one receptor.

Disease Burden and Prediction Analysis of Tracheal, Bronchus, and Lung Cancer Attributable to Residential Radon, Solid Fuels, and Particulate Matter Pollution Under Different Sociodemographic Transitions From 1990 to 2030.

BACKGROUND: This study investigated the impact of epidemiologic and sociodemographic changes in tracheal, bronchus, and lung cancer associated with residential radon, solid fuels, and particulate matter. RESEARCH QUESTION: What are the influencing factors of tracheal, bronchus, and lung cancer disease burden attributable to the three pollutants? STUDY DESIGN AND METHODS: Data were obtained from the Global Burden of Disease 2019. Age-standardized mortality rate (ASMR) and sociodemographic index (SDI) values were collected from 21 regions, and restricted cubic splines and quantile regression were used to investigate the relationship between ASMR or age-standardized disability-adjusted life years rate (ASDR), and SDI. Additionally, five countries with different SDIs were selected, and the Bayesian age-period-cohort model was used to predict the ASMR trends from 2020 to 2030. RESULTS: High SDI quintiles were associated with increased residential radon pollution. The disease burden attributed to these three pollutants was particularly severe in the middle SDI quintiles. Older adults aged 80 to 89 years had the highest age-specific mortality, and the disease burden was greater in male patients than in female patients with these cancers attributed to the pollutants. The highest ASMR attributable to particulate matter when the SDI was 0.7. As the SDI increased, the disease burden caused by radon increased, whereas the burden caused by solid fuels decreased. Projections have indicated a rise in the death burden in patients with this cancer from particulate pollution in China, India, and Uganda over the next decade. INTERPRETATION: The disease burden of tracheal, bronchus, and lung cancer attributed to the three pollutants was influenced by SDI, sex, and age. Older men are more susceptible to be affected. More preventive interventions may be required for men at younger ages to reduce the high death burden of older men. However, it is necessary to give due attention to women in specific countries in the future.

Intercellular communication in peritoneal dialysis.

Long-term peritoneal dialysis (PD) causes structural and functional alterations of the peritoneal membrane. Peritoneal deterioration and fibrosis are multicellular and multimolecular processes. Under stimulation by deleterious factors such as non-biocompatibility of PD solution, various cells in the abdominal cavity show differing characteristics, such as the secretion of different cytokines, varying protein expression levels, and transdifferentiation into other cells. In this review, we discuss the role of various cells in the abdominal cavity and their interactions in the pathogenesis of PD. An in-depth understanding of intercellular communication and inter-organ communication in PD will lead to a better understanding of the pathogenesis of this disease, enabling the development of novel therapeutic targets.

Astragalus polysaccharides augment BMSC homing via SDF-1/CXCR4 modulation: a novel approach to counteract peritoneal mesenchymal transformation and fibrosis.

PURPOSE: This study aimed to evaluate the potential of astragalus polysaccharide (APS) pretreatment in enhancing the homing and anti-peritoneal fibrosis capabilities of bone marrow mesenchymal stromal cells (BMSCs) and to elucidate the underlying mechanisms. METHODS: Forty male Sprague-Dawley rats were allocated into four groups: control, peritoneal dialysis fluid (PDF), PDF + BMSCs, and PDF + APSBMSCs (APS-pre-treated BMSCs). A peritoneal fibrosis model was induced using PDF. Dil-labeled BMSCs were administered intravenously. Post-transplantation, BMSC homing to the peritoneum and pathological alterations were assessed. Stromal cell-derived factor-1 (SDF-1) levels were quantified via enzyme-linked immunosorbent assay (ELISA), while CXCR4 expression in BMSCs was determined using PCR and immunofluorescence. Additionally, a co-culture system involving BMSCs and peritoneal mesothelial cells (PMCs) was established using a Transwell setup to examine the in vitro effects of APS on BMSC migration and therapeutic efficacy, with the CXCR4 inhibitor AMD3100 deployed to dissect the role of the SDF-1/CXCR4 axis and its downstream impacts. RESULTS: In vivo and in vitro experiments confirmed that APS pre-treatment notably facilitated the targeted homing of BMSCs to the peritoneal tissue of PDF-treated rats, thereby amplifying their therapeutic impact. PDF exposure markedly increased SDF-1 levels in peritoneal and serum samples, which encouraged the migration of CXCR4-positive BMSCs. Inhibition of the SDF-1/CXCR4 axis through AMD3100 application diminished BMSC migration, consequently attenuating their therapeutic response to peritoneal mesenchyme-to-mesothelial transition (MMT). Furthermore, APS upregulated CXCR4 expression in BMSCs, intensified the activation of the SDF-1/CXCR4 axis's downstream pathways, and partially reversed the AMD3100-induced effects. CONCLUSION: APS augments the SDF-1/CXCR4 axis's downstream pathway activation by increasing CXCR4 expression in BMSCs. This action bolsters the targeted homing of BMSCs to the peritoneal tissue and amplifies their suppressive influence on MMT, thereby improving peritoneal fibrosis.

Comparison of Secular Trends in Peptic Ulcer Diseases Mortality in China, Brazil and India during 1990-2019: An Age-Period-Cohort Analysis.

BACKGROUND: Peptic ulcer disease (PUD) is a common disease worldwide, especially in developing countries. China, Brazil, and India are among the world's fastest-growing emerging economies. This study aimed to assess long-term trends in PUD mortality and explore the effects of age, period, and cohort in China, Brazil, and India. METHODS: We collected data from the 2019 Global Burden of Disease Study and used an age-period-cohort (APC) model to estimate the effects of age, period, and cohort. We also obtained net drift, local drift, longitudinal age curve, and period/cohort rate ratios using the APC model. RESULTS: Between 1990 and 2019, the age-standardized mortality rates (ASMRs) of PUD and PUD attributable to smoking showed a downward trend in all countries and both sexes. The local drift values for both sexes of all ages were below zero, and there were obvious sex differences in net drifts between China and India. India had a more pronounced upward trend in the age effects than other countries. The period and cohort effects had a similar declining trend in all countries and both sexes. CONCLUSIONS: China, Brazil, and India had an inspiring decrease in the ASMRs of PUD and PUD attributable to smoking and to period and cohort effects during 1990-2019. The decreasing rates of Helicobacter pylori infection and the implementation of tobacco-restricting policies may have contributed to this decrease.

TMT quantitative proteomics and network pharmacology reveal the mechanism by which asiaticoside regulates the JAK2/STAT3 signaling pathway to inhibit peritoneal fibrosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese medicine, Centella asiatica (L.) Urb., has been extensively utilized in clinics to treat a variety of fibrotic disorders. Asiaticoside (ASI), as an important active ingredient, has attracted much attention in this field. However, the effect of ASI on peritoneal fibrosis (PF) is still unclear. Therefore, we evaluated the benefits of ASI for PF and mesothelial-mesenchymal transition (MMT) and revealed the underlying mechanisms. AIM OF STUDY: The objective of this investigation was to anticipate the potential molecular mechanism of ASI against peritoneal mesothelial cells (PMCs) MMT employing proteomics and network pharmacology, and to confirm it using in vivo and in vitro studies. MATERIALS AND METHODS: The mesentery of peritoneal fibrosis mice and normal mice were analyzed quantitatively for proteins that were differentially expressed using a technique tandem mass tag (TMT). Next, the core target genes of ASI against PF were screened through network pharmacology analysis, and PPI and C-P‒T networks were constructed by Cytoscape Version 3.7.2. According to the findings of a GO and KEGG enrichment analysis of differential proteins and core target genes, the signaling pathway with a high correlation degree was selected as the key signaling pathway of ASI inhibiting the PMCs MMT for further molecular docking analysis and experimental verification. RESULTS: TMT-based quantitative proteome analysis revealed the identification of 5727 proteins, of which 70 were downregulated and 178 were upregulated. Among them, the levels of STAT1, STAT2, and STAT3 in the mesentery of mice with peritoneal fibrosis were considerably lower than in the control group, indicating a role for the STAT family in the pathogenesis of peritoneal fibrosis. Then, a total of 98 ASI-PF-related targets were identified by network pharmacology analysis. JAK2 is one of the top 10 core target genes representing a potential therapeutic target. JAK/STAT signaling may represent a core pathway mediating PF effects by ASI. Molecular docking studies showed that ASI had the potential to interact favorably with target genes involved in the JAK/STAT signaling pathway, such as JAK2 and STAT3. The experimental results showed that ASI could significantly alleviate Chlorhexidine Gluconate (CG)-induced peritoneal histopathological changes and increase JAK2 and STAT3 phosphorylation levels. In TGF-ß1-stimulated HMrSV5 cells, E-cadherin expression levels were dramatically reduced whereas Vimentin, p-JAK2, α-SMA, and p-STAT3 expression levels were considerably increased. ASI inhibited the TGF-ß1-induced HMrSV5 cell MMT, decreased the activation of JAK2/STAT3 signaling, and increased the nuclear translocation of p-STAT3, which was consistent with the effect of the JAK2/STAT3 pathway inhibitor AG490. CONCLUSION: ASI can inhibit PMCs MMT and alleviate PF by regulating the JAK2/STAT3 signaling pathway.

Causal Effects of Specific Gut Microbiota on Chronic Kidney Diseases and Renal Function-A Two-Sample Mendelian Randomization Study.

BACKGROUND: Targeting the gut microbiota may become a new therapeutic to prevent and delay the progression of chronic kidney disease (CKD). Nonetheless, the causal relationship between specific intestinal flora and CKD is still unclear. MATERIALS AND METHOD: To identify genetically predicted microbiota, we used summary data from genome-wide association studies on gut microbiota in 18340 participants from 24 cohorts. Furthermore, we genetically predicted the causal relationship between 211 gut microbiotas and six phenotypes (outcomes) (CKD, estimated glomerular filtration rate (eGFR), urine albumin to creatinine ratio (UACR), dialysis, rapid progress to CKD, and rapid decline of eGFR). Four Mendelian randomization (MR) methods, including inverse variance weighted (IVW), MR-Egger, weighted median, and weighted mode were used to investigate the casual relationship between gut microbiotas and various outcomes. The result of IVW was deemed as the primary result. Then, Cochrane's Q test, MR-Egger, and MR-PRESSO Global test were used to detect heterogeneity and pleiotropy. The leave-one method was used for testing the stability of MR results and Bonferroni-corrected was used to test the strength of the causal relationship between exposure and outcome. RESULTS: Through the MR analysis of 211 microbiotas and six clinical phenotypes, a total of 36 intestinal microflora were found to be associated with various outcomes. Among them, Class Bacteroidia (=-0.005, 95% CI: -0.001 to -0.008, p = 0.002) has a strong causality with lower eGFR after the Bonferroni-corrected test, whereas phylum Actinobacteria (OR = 1.0009, 95%CI: 1.0003-1.0015, p = 0.0024) has a strong causal relationship with dialysis. The Cochrane's Q test reveals that there is no significant heterogeneity between various single nucleotide polymorphisms. In addition, no significant level of pleiotropy was found according to MR-Egger and MR-PRESSO Global tests. CONCLUSIONS: Through the two-sample MR analysis, we identified the specific intestinal flora that has a causal relationship with the incidence and progression of CKD at the level of gene prediction, which may provide helpful biomarkers for early disease diagnosis and potential therapeutic targets for CKD.

Excitatory somatostatin interneurons in the dentate gyrus drive a widespread seizure network in cortical dysplasia.

Seizures due to cortical dysplasia are notorious for their poor prognosis even with medications and surgery, likely due to the widespread seizure network. Previous studies have primarily focused on the disruption of dysplastic lesions, rather than remote regions such as the hippocampus. Here, we first quantified the epileptogenicity of the hippocampus in patients with late-stage cortical dysplasia. We further investigated the cellular substrates leading to the epileptic hippocampus, using multiscale tools including calcium imaging, optogenetics, immunohistochemistry and electrophysiology. For the first time, we revealed the role of hippocampal somatostatin-positive interneurons in cortical dysplasia-related seizures. Somatostatin-positive were recruited during cortical dysplasia-related seizures. Interestingly, optogenetic studies suggested that somatostatin-positive interneurons paradoxically facilitated seizure generalization. By contrast, parvalbumin-positive interneurons retained an inhibitory role as in controls. Electrophysiological recordings and immunohistochemical studies revealed glutamate-mediated excitatory transmission from somatostatin-positive interneurons in the dentate gyrus. Taken together, our study reveals a novel role of excitatory somatostatin-positive neurons in the seizure network and brings new insights into the cellular basis of cortical dysplasia.

The role of Na+ -K+ -ATPase in the epileptic brain.

Na+-K+-ATPase, a P-type ATP-powered ion transporter on cell membrane, plays a vital role in cellular excitability. Cellular hyperexcitability, accompanied by hypersynchronous firing, is an important basis for seizures/epilepsy. An increasing number of studies point to a significant contribution of Na+-K+-ATPase to epilepsy, although discordant results exist. In this review, we comprehensively summarize the structure and physiological function of Na+-K+-ATPase in the central nervous system and critically evaluate the role of Na+-K+-ATPase in the epileptic brain. Importantly, we further provide perspectives on some possible research directions and discuss its potential as a therapeutic target for the treatment of epilepsy.

Trends of ischemic heart disease mortality attributable to household air pollution during 1990-2019 in China and India: an age-period-cohort analysis.

Cardiovascular diseases (CVD) caused by household air pollution (HAP) have sparked widespread concern globally in the recent decade. Meanwhile, increased ischemic heart disease (IHD) mortality has been the leading cause of worldwide CVD deaths. Both China and India experienced a high IHD burden and high exposure to HAP. The present study aimed to estimate and compare the long-term trends of HAP-attributable IHD mortality in the two countries. The data of this study were extracted from the Global Burden of Diseases (GBD) Study 2019. The age-period-cohort (APC) analysis was utilized to estimate the independent trends of the age, period, and cohort effects from 1990 to 2019. The age-standardized mortality rates (ASMRs) of HAP-attributable IHD have fallen faster in China than in India for both sexes. The local drift and net drift values were < 0 for all age groups in both countries. The age effects in both countries and sexes increased with time, suggesting age is a risk factor for IHD; conversely, period and cohort effects in China demonstrated a faster decline in both genders than in India. It indicated that China has been more successful than India in decreasing HAP-attributable IHD mortality.

Comparison of Secular Trends in Esophageal Cancer Mortality in China and Japan during 1990-2019: An Age-Period-Cohort Analysis.

Esophageal cancer is a prevalent and often fatal malignancy all over the world, with China and Japan bearing a disproportionately high burden. Consequently, we explored and compared the long-term changes in esophageal cancer mortality in China and Japan from 1990 to 2019 to see if there were any etiological clues. From 1990 to 2019, data on mortality in China and Japan were gathered from the Global Burden of Disease Study 2019 (GBD 2019). The age-period-cohort (APC) model was utilized to evaluate the effects of age, period, and cohort. Between 1990 and 2019, the age-standardized mortality rates (ASMRs) for esophageal cancer fell in both nations, with China showing a tremendous reduction after 2005. The overall net drifts per year were more impressive in China (-5.22% [95% CI, -5.77 to -4.68] for females, -1.98% [-2.22 to -1.74] for males) than in Japan (-0.50% [-0.91 to -0.08] for females, -1.86% [-2.12 to -1.59] for males), and the local drift values in both countries were less than zero in all age groups for both sexes. The longitudinal age curves of esophageal cancer mortality increased as age advances and the sex disparity gradually exacerbates with age. The period and cohort effects were uncovered to have similar declining patterns for both sexes in both nations; however, the improvement of cohort effects for China's younger generation has stagnated. The ASMRs, period effects, and cohort effects have decreased for both countries and sexes over the 1990-2019 period. The decline in cohort effects for China's younger generation has plateaued, possibly due to the rising rates of smoking and obesity among Chinese youngsters. Comprehensive population-level treatments aimed at smoking cessation, obesity prevention, and gastrointestinal endoscopy screening should be carried out immediately, particularly for men and older birth cohorts at a higher risk of esophageal cancer.

Long-term trends in the incidence of depressive disorders in China, the United States, India and globally: A comparative study from 1990 to 2019.

Background: Depressive disorders have become an increasingly significant public health issue. This study is intended to show the trend of the incidence of depressive disorders in China, the United States, India and the world from 1990 to 2019, as well as the impact of age, period and cohort on it. Methods: Extracting incidence data from the Global Burden of Disease Study 2019, we determined trends in the age-standardized incidence rate (ASIR) using Joinpoint regression. An age-period-cohort analysis was implemented to describe the effects of age, period, and cohort, as well as the long-term tendencies. Results: From 1990 to 2019, the ASIR of depressive disorders in China was lower than that in the United States; India is lower than the United States in the first 5 years, showing a downward trend. The incidence in India and the United States is higher than the global average. The ASIR of women in the three countries is higher than that of men. In China, the elderly, early period and people born around 1954 have a higher risk of depressive disorders. In the United States, young people born around 1999 have a higher risk of depressive disorders. India is similar to China. Conclusion: From 1990 to 2019, the age effect of China as a whole increased, and the period became stable, and the cohort effect declined. The overall age and period effects of the United States reduced, while the cohort effect increased. The age effect in India increased, while the period and cohort effects decreased. Depressive disorders are becoming ever more serious worldwide, and we'd better take measures to reduce its incidence according to the cohort effect of each age group.

Activated astrocytes attenuate neocortical seizures in rodent models through driving Na+-K+-ATPase.

Epileptic seizures are widely regarded to occur as a result of the excitation-inhibition imbalance from a neuro-centric view. Although astrocyte-neuron interactions are increasingly recognized in seizure, elementary questions about the causal role of astrocytes in seizure remain unanswered. Here we show that optogenetic activation of channelrhodopsin-2-expressing astrocytes effectively attenuates neocortical seizures in rodent models. This anti-seizure effect is independent from classical calcium signaling, and instead related to astrocytic Na+-K+-ATPase-mediated buffering K+, which activity-dependently inhibits firing in highly active pyramidal neurons during seizure. Compared with inhibition of pyramidal neurons, astrocyte stimulation exhibits anti-seizure effects with several advantages, including a wider therapeutic window, large-space efficacy, and minimal side effects. Finally, optogenetic-driven astrocytic Na+-K+-ATPase shows promising therapeutic effects in a chronic focal cortical dysplasia epilepsy model. Together, we uncover a promising anti-seizure strategy with optogenetic control of astrocytic Na+-K+-ATPase activity, providing alternative ideas and a potential target for the treatment of intractable epilepsy.

3D-Printed Micro-Tweezers with a Compliant Mechanism Designed Using Topology Optimization.

The development of handling technology for microscopic biological samples such as cells and spheroids has been required for the advancement of regenerative medicine and tissue engineering. In this study, we developed micro-tweezers with a compliant mechanism to manipulate organoids. The proposed method combines high-resolution microstereolithography that uses a blue laser and topology optimization for shape optimization of micro-tweezers. An actuation system was constructed using a linear motor stage with a force control system to operate the micro-tweezers. The deformation of the topology-optimized micro-tweezers was examined analytically and experimentally. The results verified that the displacement of the tweezer tip was proportional to the applied load; furthermore, the displacement was sufficient to grasp biological samples with an approximate diameter of several hundred micrometers. We experimentally demonstrated the manipulation of an organoid with a diameter of approximately 360 µm using the proposed micro-tweezers. Thus, combining microstereolithography and topology optimization to fabricate micro-tweezers can be potentially used in modifying tools capable of handling various biological samples.