ATF2/BAP1 Axis Mediates Neuronal Apoptosis After Subarachnoid Hemorrhage via P53 Pathway.

BACKGROUND: Neuronal apoptosis plays an essential role in the pathogenesis of brain injury after subarachnoid hemorrhage (SAH). BAP1 (BRCA1-associated protein 1) is considered to exert pro-apoptotic effects in multiple diseases. However, evidence supporting the effect of BAP1 on the apoptotic response to SAH is lacking. Therefore, we aimed to confirm the role of BAP1 in SAH-induced apoptosis. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to detect BAP1 expression in the cerebrospinal fluid. Endovascular perforation was performed in mice to induce SAH. Lentiviral short hairpin RNA targeting BAP1 mRNA was transduced into the ipsilateral cortex of mice with SAH to investigate the role of BAP1 in neuronal damage. Luciferase and coimmunoprecipitation assays were performed to investigate the mechanism through which BAP1 participates in hemin-induced SAH. RESULTS: First, BAP1 expression was upregulated in the cerebrospinal fluid of patients with SAH and positively associated with unfavorable outcomes. ATF2 (activating transcription factor-2) then regulated BAP1 expression by binding to the BAP1 promoter. In addition, BAP1 overexpression enhanced P53 activity and stability by reducing P53 proteasome-mediated degradation. Subsequently, elevated P53 promoted neuronal apoptosis via the P53 pathway. Inhibition of the neuronal BAP1/P53 axis significantly reduced neurological deficits and neuronal apoptosis and improved neurological dysfunction in mice after SAH. CONCLUSIONS: Our results suggest that the neuronal ATF2/BAP1 axis exerts a brain-damaging effect by modulating P53 activity and stability and may be a novel therapeutic target for SAH.

System-wide identification of novel de-ubiquitination targets for USP10 in gastric cancer metastasis through multi-omics screening.

OBJECTIVE: Ubiquitin-specific peptidase 10 (USP10), a typical de-ubiquitinase, has been found to play a double-edged role in human cancers. Previously, we reported that the expression of USP10 was negatively correlated with the depth of gastric wall invasion, lymph node metastasis, and prognosis in gastric cancer (GC) patients. However, it remains unclear whether USP10 can regulate the metastasis of GC cells through its de-ubiquitination function. METHODS: In this study, proteome, ubiquitinome, and transcriptome analyses were conducted to comprehensively identify novel de-ubiquitination targets for USP10 in GC cells. Subsequently, a series of validation experiments, including in vitro cell culture studies, in vivo metastatic tumor models, and clinical sample analyses, were performed to elucidate the regulatory mechanism of USP10 and its de-ubiquitination targets in GC metastasis. RESULTS: After overexpression of USP10 in GC cells, 146 proteins, 489 ubiquitin sites, and 61 mRNAs exhibited differential expression. By integrating the results of multi-omics, we ultimately screened 9 potential substrates of USP10, including TNFRSF10B, SLC2A3, CD44, CSTF2, RPS27, TPD52, GPS1, RNF185, and MED16. Among them, TNFRSF10B was further verified as a direct de-ubiquitination target for USP10 by Co-IP and protein stabilization assays. The dysregulation of USP10 or TNFRSF10B affected the migration and invasion of GC cells in vitro and in vivo models. Molecular mechanism studies showed that USP10 inhibited the epithelial-mesenchymal transition (EMT) process by increasing the stability of TNFRSF10B protein, thereby regulating the migration and invasion of GC cells. Finally, the retrospective clinical sample studies demonstrated that the downregulation of TNFRSF10B expression was associated with poor survival among 4 of 7 GC cohorts, and the expression of TNFRSF10B protein was significantly negatively correlated with the incidence of distant metastasis, diffuse type, and poorly cohesive carcinoma. CONCLUSIONS: Our study established a high-throughput strategy for screening de-ubiquitination targets for USP10 and further confirmed that inhibiting the ubiquitination of TNFRSF10B might be a promising therapeutic strategy for GC metastasis.

Incidence and risk factors of stress urinary incontinence after laparoscopic hysterectomy.

OBJECTIVE: To observe the long-term effects of total hysterectomy on urinary function, evaluate the effects of preoperative nutritional status, urinary occult infection, and surgical factors on the induction of postoperative stress urinary incontinence (SUI), and explore the incidence and risk factors of SUI. STUDY DESIGN: From January 2017 to December 2017, 164 patients with benign non-prolapsing diseases who underwent a laparoscopic total hysterectomy in the First People's Hospital of Taicang were selected as the analysis objects. The International Incontinence Standard Questionnaire for Female Lower Urinary Tract Symptoms (ICIQ-FLUTS) and Pelvic Floor Impact Questionnaire-short version 20 (PFDI-20) were used for telephone follow-up to subjectively assess the urinary function of patients, collect their medical records, and statistically analyze the number of postoperative SUI cases. Logistic multivariate analysis was used to analyze the influencing factors of postoperative female SUI, presented as adjusted odds ratios with 95% confidence intervals. RESULTS: Only 97 out of 164 patients completed the ICIQ-FLUTS and PFDI-20 questionnaires. Among these participants, 28 patients (28.86%) were diagnosed with SUI (study group), while 69 patients (71.13%) were classified as women without SUI (control group). The age, menopause, parity ≥ 2 times, Body mass index (BMI) ≥ 28 kg/m2, neonatal weight ≥ 4000 g, history of chronic cough, preoperative hemoglobin ≤ 100 g/L, preoperative urine bacteria ≥ 100u/L, preoperative uterine volume ≥ 90 cm3, intraoperative blood loss, and operation time of the study group were compared with those of the control group. The differences were statistically significant (P < 0.05). Further Logistic multivariate analysis showed that menopause, preoperative hemoglobin ≤ 100 g/L, preoperative urine bacteria ≥ 100u/L, uterine volume ≥ 90 cm3, history of chronic cough, BMI ≥ 28 kg/m2 were risk factors for postoperative SUI in patients undergoing hysterectomy (P < 0.05). CONCLUSIONS: Hysterectomy for benign non-prolapse diseases has a long-term potential impact on the urinary system of patients, and the risk of postoperative SUI increases. The main risk factors of SUI are parity, menopausal status, obesity, preoperative nutritional status, and occult infection of the urinary system.

Does social capital aid in leveling the income gradient in child mental health? A structural analysis of the left-behind and not-left-behind Chinese children.

BACKGROUND: Few prior studies have investigated the income gradient in child mental health from a socio-environmental perspective. In an age when child mental health problems in a rapidly changing social environment have become a worldwide issue, an understanding of the socio-environmental mechanisms of the income disparities in child mental health outcomes is imperative and cost-effective. METHODS: By conducting structural equation analyses with Chinese nationally representative survey data, this study explored the family income gradient in child depression and its potential socio-environmental pathways at the neighborhood, family and school levels, differentiating left-behind and not-left-behind children. RESULTS: We found a robust family income gradient in depressive symptoms. Neighborhood cohesion mitigated the income gradient in depressive symptoms by playing a suppression role. School social capital acted as a mediator. Neighborhood trust, neighborhood safety and family social capital played no significant impact. The mitigating and mediating roles of social capital components were significant among only the not-left-behind children. CONCLUSIONS: To reduce income-related inequalities in child mental health in the long run, integrating policies that directly reduce poverty with policies that improve distal socio-environments is necessary.

Different gender-derived gut microbiota influence stroke outcomes by mitigating inflammation.

BACKGROUND AND PURPOSE: Stroke is associated with high disability and mortality rates and increases the incidence of organ-related complications. Research has revealed that the outcomes and prognosis of stroke are regulated by the state of the intestinal microbiota. However, the possibility that the manipulation of the intestinal microbiota can alter sex-related stroke outcomes remain unknown. METHODS: To verify the different effects of microbiota from different sexes on stroke outcomes, we performed mouse fecal microbiota transplantation (FMT) and established a model of ischemic stroke. Male and female mice received either male or female microbiota through FMT. Ischemic stroke was triggered by MCAO (middle cerebral artery occlusion), and sham surgery served as a control. Over the next few weeks, the mice underwent neurological evaluation and metabolite and inflammatory level detection, and we collected fecal samples for 16S ribosomal RNA analysis. RESULTS: We found that when the female mice were not treated with FMT, the microbiota (especially the Firmicutes-to-Bacteroidetes ratio) and the levels of three main metabolites tended to resemble those of male mice after experimental stroke, indicating that stroke can induce an ecological imbalance in the biological community. Through intragastric administration, the gut microbiota of male and female mice was altered to resemble that of the other sex. In general, in female mice after MCAO, the survival rate was increased, the infarct area was reduced, behavioral test performance was improved, the release of beneficial metabolites was promoted and the level of inflammation was mitigated. In contrast, mice that received male microbiota were much more hampered in terms of protection against brain damage and the recovery of neurological function. CONCLUSION: A female-like biological community reduces the level of systemic proinflammatory cytokines after ischemic stroke. Poor stroke outcomes can be positively modulated following supplementation with female gut microbiota.

First Report of Clonostachys rosea Causing rot of Morchella sextelata in Anhui Province, China.

Morel mushroom (Morchella spp.) is a valuable mushroom, which has extremely high nutritional and economic value. In the early March of 2022, a serious rot disease was observed on approximately 30% fruiting bodies at an M. esculenta farm of Suzhou City, Anhui province, China. A white mold-like hyphae was initially present on the pileus, which then gradually spread to the whole fruiting body, eventually resulting in softening of the fruiting body and death. This disease developed rapidly at relatively high temperature (>20°C) and humidity (>85%), resulting in approximately 80% loss of yield. Twenty infected tissues were cut into small pieces (5 × 5 mm) and placed on potato dextrose agar (PDA) and incubated at 25°C for 5 days. Fifteen morphologically similar isolates were obtained and purified using the single spore isolation technique. Colonies of these isolates were yellowish-white, and tomentose with thick aerial hyphae after 7 days at 25°C on PDA plates. Conidiophores were dimorphic: primary conidiophores were Verticillium-like, secondary conidiophores were penicillate. Primary conidiophore stipe length ranged from 68.6 to 180.5 µm, and the base width was 3.2-5.6 µm. Phialides were solitary, straight, generally slightly tapering towards the tip, each producing a small, hyaline drop of conidia. Secondary conidiophores stipe length ranged from 68.4 to 120.5 µm, the base measured 3.3-6.1µm. Phialides were straight to slightly curved, slightly flask-shaped, with widest point below the middle, slightly tapering in the upper part, without visible collarette. Conidia were colorless, smooth, slightly curved, and distally broadly rounded with an average size of 6.3 to 8.2 × 2.4 to 3.7 µm (n=30). These isolates were initially identified as Clonostachys rosea based on morphological features (Schroers et al. 1999). To confirm the identity of C. rosea, primers ITS1/ITS4 (White et al. 1990) and EF1-728F/ EF1-986R (Carbone and Kohn 1999), were used to amplify the internal transcribed spacer (ITS), and translation elongation factor 1-alpha (EF-1α) genes of the representative isolate 5-3-2. These sequences were deposited in GenBank (GenBank accession nos. ON614093 and ON630916) and had 100% and 99.45% nucleotide identity with Clonostachys rosea E5R(17) and Clonostachys rosea KGSJ26 (GenBank accession nos. MK752437 and MT462122), respectively. Single conidium were isolated and multiplied on PDA for pathogenicity testing. To fulfill Koch's postulates, pathogenicity tests were performed using the fruiting bodies. Spores of C. rosea isolates 5-3-2 were collected and diluted with sterile distilled water at a concentration of 1 × 106 conidia/ml. Five healthy fruiting bodies were inoculated with 1 ml of the spore suspension, which were maintained in an artificial climate chamber at 22°C and 85% humidity. Sterile water inoculated on other 10 healthy fruiting bodies served as control. Mycelia grew rapidly and overgrew more than half of the fruiting bodies within 3 days. The fruiting bodies died five days after inoculation. Clonostachys rosea isolates were re-isolated from symptomatic fruiting bodies and identified by the methods described above. The control group showed no symptoms. The experiment was conducted twice. To our knowledge, this is the first identification of Clonostachys rosea as the causal agent of the Morchella sextelata rot.

Application of a Bayesian Network Based on Multi-Source Information Fusion in the Fault Diagnosis of a Radar Receiver.

A radar is an important part of an air defense and combat system. It is of great significance to military defense to improve the effectiveness of radar state monitoring and the accuracy of fault diagnosis during operation. However, the complexity of radar equipment's structure and the uncertainty of the operating environment greatly increase the difficulty of fault diagnosis in real life situations. Therefore, a Bayesian network diagnosis method based on multi-source information fusion technology is proposed to solve the fault diagnosis problems caused by uncertain factors such as the high integration and complexity of the system during the process of fault diagnosis. Taking a fault of a radar receiver as an example, we study 2 typical fault phenomena and 21 fault points. After acquiring and processing multi-source information, establishing a Bayesian network model, determining conditional probability tables (CPTs), and finally outputting the diagnosis results. The results are convincing and consistent with reality, which verifies the effectiveness of this method for fault diagnosis in radar receivers. It realizes device-level fault diagnosis, which shortens the maintenance time for radars and improves the reliability and maintainability of radars. Our results have significance as a guide for judging the fault location of radars and predicting the vulnerable components of radars.

Relevant mediators involved in and therapies targeting the inflammatory response induced by activation of the NLRP3 inflammasome in ischemic stroke.

The nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family pyrin domain-containing 3 (NLRP3) inflammasome is a member of the NLR family of inherent immune cell sensors. The NLRP3 inflammasome can detect tissue damage and pathogen invasion through innate immune cell sensor components commonly known as pattern recognition receptors (PRRs). PRRs promote activation of nuclear factor kappa B (NF-κB) pathways and the mitogen-activated protein kinase (MAPK) pathway, thus increasing the transcription of genes encoding proteins related to the NLRP3 inflammasome. The NLRP3 inflammasome is a complex with multiple components, including an NAIP, CIITA, HET-E, and TP1 (NACHT) domain; apoptosis-associated speck-like protein containing a CARD (ASC); and a leucine-rich repeat (LRR) domain. After ischemic stroke, the NLRP3 inflammasome can produce numerous proinflammatory cytokines, mediating nerve cell dysfunction and brain edema and ultimately leading to nerve cell death once activated. Ischemic stroke is a disease with high rates of mortality and disability worldwide and is being observed in increasingly younger populations. To date, there are no clearly effective therapeutic strategies for the clinical treatment of ischemic stroke. Understanding the NLRP3 inflammasome may provide novel ideas and approaches because targeting of upstream and downstream molecules in the NLRP3 pathway shows promise for ischemic stroke therapy. In this manuscript, we summarize the existing evidence regarding the composition and activation of the NLRP3 inflammasome, the molecules involved in inflammatory pathways, and corresponding drugs or molecules that exert effects after cerebral ischemia. This evidence may provide possible targets or new strategies for ischemic stroke therapy.

The immune response of T cells and therapeutic targets related to regulating the levels of T helper cells after ischaemic stroke.

Through considerable effort in research and clinical studies, the immune system has been identified as a participant in the onset and progression of brain injury after ischaemic stroke. Due to the involvement of all types of immune cells, the roles of the immune system in stroke pathology and associated effects are complicated. Past research concentrated on the functions of monocytes and neutrophils in the pathogenesis of ischaemic stroke and tried to demonstrate the mechanisms of tissue injury and protection involving these immune cells. Within the past several years, an increasing number of studies have elucidated the vital functions of T cells in the innate and adaptive immune responses in both the acute and chronic phases of ischaemic stroke. Recently, the phenotypes of T cells with proinflammatory or anti-inflammatory function have been demonstrated in detail. T cells with distinctive phenotypes can also influence cerebral inflammation through various pathways, such as regulating the immune response, interacting with brain-resident immune cells and modulating neurogenesis and angiogenesis during different phases following stroke. In view of the limited treatment options available following stroke other than tissue plasminogen activator therapy, understanding the function of immune responses, especially T cell responses, in the post-stroke recovery period can provide a new therapeutic direction. Here, we discuss the different functions and temporal evolution of T cells with different phenotypes during the acute and chronic phases of ischaemic stroke. We suggest that modulating the balance between the proinflammatory and anti-inflammatory functions of T cells with distinct phenotypes may become a potential therapeutic approach that reduces the mortality and improves the functional outcomes and prognosis of patients suffering from ischaemic stroke.

Extracellular vesicle-derived miRNA as a novel regulatory system for bi-directional communication in gut-brain-microbiota axis.

The gut-brain-microbiota axis (GBMAx) coordinates bidirectional communication between the gut and brain, and is increasingly recognized as playing a central role in physiology and disease. MicroRNAs are important intracellular components secreted by extracellular vesicles (EVs), which act as vital mediators of intercellular and interspecies communication. This review will present current advances in EV-derived microRNAs and their potential functional link with GBMAx. We propose that EV-derived microRNAs comprise a novel regulatory system for GBMAx, and a potential novel therapeutic target for modifying GBMAx in clinical therapy.

Does social capital interact with economic hardships in influencing older adults' health? A study from China.

BACKGROUND: The importance of social and economic capital as predictors of health is widely documented, yet the complexity of interactions between them and effects on older people's health is still unclear. Combining the material and psychosocial explanations of health, this study explores the potential interactions between social and economic capital in influencing older adults' health in urban and rural China. METHODS: Using data from the China Family Panel Survey, physical and mental health in 2018 were regressed on social and economic capital indicators in 2016, controlling for sociodemographic characteristics of 3535 respondents aged 65 and older. Rothman's synergy index was calculated to investigate potential interaction effects. RESULTS: Economic hardships were significantly related to both self-reported health and mental health. Neighborhood cohesion and social participation were significantly associated with mental health for all, bonding trust was significantly associated with mental health for urban older people. We found no significant associations between social capital components and self-reported health. There was an interaction effect between low neighborhood cohesion and economic hardships, and between low social participation and economic hardships, creating an increased burden of poor mental health. The interaction effect between low bonding trust and economic hardships on mental health was apparent only among urban older people. CONCLUSIONS: Geographical settings are important factors in the complexity between social and economic capital in affecting older health. Intervention efforts directed towards reducing simultaneously multiple dimensions of deprivation, such as poverty, social exclusion, social isolation, could be helpful in improving older people's health. In materially deprived places, policies to promote health equity by improving social capital but without eliminating poverty may be less effective.

On the phase fading effect in the dual-pulse heterodyne demodulated distributed acoustic sensing system.

Understanding signal fading effect is essential for the application of Rayleigh-scattering-based distributed acoustic fibre sensors (DASs) due to the nature of coherent beam interference within the pulse length. Statistical properties for the intensity of the Rayleigh backscattered light (i.e. intensity fading) and its impact on the sensitivity of DAS systems have been intensely studied over the last decades. Here we for the first time establish an analytical model for the phase signal retrieved from the dual-pulse heterodyne demodulated DAS system, which can be exploited to investigate the phase fading effect in this system. The developed model reveals that the phase fading phenomenon mainly originates from the randomness in the phase retardant of the Rayleigh scatters. The quantitatively resolved statistical features of the phase fading is confirmed by experimental results. Based on the analytical model, a noise figure is defined to characterize the global fading-induced noise level via taking into account contributions from all channels along the sensing fiber. The model also reproduces the anti-correlation relation between the power spectrum density of retrieved phase at the heterodyne frequency and the phase fading noise level. Following the analysis and the definition of the noise figure, an optimized real-time weighted-channel stack algorithm is developed to efficiently suppress the fading noise. Experimental results show that the algorithm can achieve a maximum noise figure reduction of 15.8 dB without increasing the system complexity.

Performance Improvement of Dual-Pulse Heterodyne Distributed Acoustic Sensor for Sound Detection.

Phase fading is fatal to the performance of distributed acoustic sensors (DASs) influencing its capability of distributed measurement as well as its noise level. Here, we report the experimental observation of a strong negative correlation between the relative power spectrum density (PSD) at the heterodyne frequency and the noise floor of the detected phase for the heterodyne demodulated distributed acoustic sensor (HD-DAS) system. We further propose a weighted-channel stack algorithm (WCSA) to alleviate the phase fading noise via an enhancement of the relative PSD at the heterodyne frequency. Experimental results show that the phase noise of the demodulated signal can be suppressed by 13.7 dB under optimal condition. As a potential application, we exploited the improved HD-DAS system to retrieve a piece of music lasted for 205 s, demonstrating the reliability of detecting wideband sound signal without distortion.

Bidirectional gut-brain-microbiota axis as a potential link between inflammatory bowel disease and ischemic stroke.

Emerging evidence suggests that gut-brain-microbiota axis (GBMAx) may play a pivotal role linking gastrointestinal and neuronal disease. In this review, we summarize the latest advances in studies of GBMAx in inflammatory bowel disease (IBD) and ischemic stroke. A more thorough understanding of the GBMAx could advance our knowledge about the pathophysiology of IBD and ischemic stroke and help to identify novel therapeutic targets via modulation of the GBMAx.

Distributed gas sensing with optical fibre photothermal interferometry.

We report the first distributed optical fibre trace-gas detection system based on photothermal interferometry (PTI) in a hollow-core photonic bandgap fibre (HC-PBF). Absorption of a modulated pump propagating in the gas-filled HC-PBF generates distributed phase modulation along the fibre, which is detected by a dual-pulse heterodyne phase-sensitive optical time-domain reflectometry (OTDR) system. Quasi-distributed sensing experiment with two 28-meter-long HC-PBF sensing sections connected by single-mode transmission fibres demonstrated a limit of detection (LOD) of ∼10 ppb acetylene with a pump power level of 55 mW and an effective noise bandwidth (ENBW) of 0.01 Hz, corresponding to a normalized detection limit of 5.5ppb⋅W/Hz. Distributed sensing experiment over a 200-meter-long sensing cable made of serially connected HC-PBFs demonstrated a LOD of ∼ 5 ppm with 62.5 mW peak pump power and 11.8 Hz ENBW, or a normalized detection limit of 312ppb⋅W/Hz. The spatial resolution of the current distributed detection system is limited to ∼ 30 m, but it is possible to reduce down to 1 meter or smaller by optimizing the phase detection system.

Multi-event waveform-retrieved distributed optical fiber acoustic sensor using dual-pulse heterodyne phase-sensitive OTDR.

We demonstrate a novel type of distributed optical fiber acoustic sensor, with the ability to detect and retrieve actual temporal waveforms of multiple vibration events that occur simultaneously at different positions along the fiber. The system is realized via a dual-pulse phase-sensitive optical time-domain reflectometry, and the actual waveform is retrieved by heterodyne phase demodulation. Experimental results show that the system has a background noise level as low as 8.91×10-4 rad/√Hz with a demodulation signal-to-noise ratio of 49.17 dB at 1 kHz, and can achieve a dynamic range of ∼60 dB at 1 kHz (0.1 to 104 rad) for phase demodulation, as well as a detection frequency range from 20 Hz to 25 kHz.

Effects of Passive Smoking on Glycemic Parameters and Lipid Profiles in a Chinese Female Population.

BACKGROUND: The adverse effects of exposure to second-hand cigarette smoke on glucose and lipid parameters in women and the underlying mechanisms are not well understood. In this prospective community-based study, we examined the effects of passive smoking on glycemic parameters and lipid profiles in a Chinese female population. METHODS: Of 3197 healthy women enrolled (30 - 75 years), 2082 self-reported passive exposure to smoke (≥ 15 minutes/day, 3 days/week, > 1 year) and 1115 had no smoke exposure (control group). Data was collected via questionnaire, body measurements, and laboratory assays for glycemic parameters and lipid profiles. RESULTS: (1) Women exposed to second-hand smoke had significantly higher fasting plasma glucose, hemoglobin A1c, total cholesterol, triglycerides, low-density lipoprotein cholesterol, and waist-to-hip ratio compared with the control group (p < 0.05), while 2-h plasma glucose, high-density lipoprotein cholesterol levels, and body mass indexes were similar compared with the control group (p > 0.05). (2) Logistic regression analysis showed that after adjusting for potential confounders, passive smoking women increased the risks of hemoglobin A1c and total cholesterol (p < 0.05). CONCLUSIONS: In this population of Chinese women, exposure to second-hand cigarette smoke was associated with adverse effects on glucose and lipid profiles, suggesting an increased risk of diabetes and cardiovascular disease. These findings support the benefit of stopping smoking in the home and implementing no-smoking regulations in public areas in China and other developing countries to prevent diabetes and other chronic diseases.

Meisoindigo, but not its core chemical structure indirubin, inhibits zebrafish interstitial leukocyte chemotactic migration.

CONTEXT: Inflammatory disease is a big threat to human health. Leukocyte chemotactic migration is required for efficient inflammatory response. Inhibition of leukocyte chemotactic migration to the inflammatory site has been shown to provide therapeutic targets for treating inflammatory diseases. OBJECTIVE: Our study was designed to discover effective and safe compounds that can inhibit leukocyte chemotactic migration, thus providing possible novel therapeutic strategy for treating inflammatory diseases. MATERIALS AND METHODS: In this study, we used transgenic zebrafish model (Tg:zlyz-EGFP line) to visualize the process of leukocyte chemotactic migration. Then, we used this model to screen the hit compound and evaluate its biological activity on leukocyte chemotactic migration. Furthermore, western blot analysis was performed to evaluate the effect of the hit compound on the AKT or ERK-mediated pathway, which plays an important role in leukocyte chemotactic migration. RESULTS: In this study, using zebrafish-based chemical screening, we identified that the hit compound meisoindigo (25 µM, 50 µM, 75 µM) can significantly inhibit zebrafish leukocyte chemotactic migration in a dose-dependent manner (p = 0.01, p = 0.0006, p < 0.0001). Also, we found that meisoindigo did not affect the process of leukocyte reverse migration (p = 0.43). Furthermore, our results unexpectedly showed that indirubin, the core structure of meisoindigo, had no significant effect on zebrafish leukocyte chemotactic migration (p = 0.6001). Additionally, our results revealed that meisoindigo exerts no effect on the Akt or Erk-mediated signalling pathway. DISCUSSION AND CONCLUSION: Our results suggest that meisoindigo, but not indirubin, is effective for inhibiting leukocyte chemotactic migration, thus providing a potential therapeutic agent for treating inflammatory diseases.

Glycyrrhizin protects against focal cerebral ischemia via inhibition of T cell activity and HMGB1-mediated mechanisms.

BACKGROUND: Glycyrrhizin (Gly) protects against brain injury induced by stroke. We studied whether Gly achieves its protection by inhibiting T cell activity and high-mobility group box 1 (HMGB1) release in the ischemic brain. METHODS: Stroke was induced by transient middle cerebral artery occlusion in rats and mice. Gly was injected intraperitoneally before or after stroke. We measured infarction, neuroinflammatory cells, gene expressions of interferon-γ (IFNγ), IL-4, and IL-10 in CD4 T cells, HMGB1 release, and T cell proliferation in cultured splenocytes. RESULTS: Gly treatment reduced infarctions and neuroinflammation characterized by the infiltration of CD68-positive macrophages and myeloperoxidase-positive neutrophils, which corresponds to a reduction in the number of T cells and their subsets, CD4 and CD8 T cells, in the ischemic brain, as measured by flow cytometry. Unlike in wild-type animals, Gly did not offer protection in nude rats and severe combined immunodeficient (SCID) mice who had no T cells, while Gly reduced infarction in both nude rats and SCID mice whose T cells were reconstituted, suggesting that T cells should be the target of Gly. In addition, Gly administration inhibited T cell proliferation stimulated by ConA in in vitro assays and inhibited HMGB1 release from the ischemic brain. Furthermore, Gly attenuated gene expression of IFNγ, but not IL-4 and IL-10 in CD4 T cells. Lastly, HMGB1 promoted T cell proliferation stimulated by ConA, which was inhibited by the addition of Gly. CONCLUSIONS: Gly blocks infarction by inhibiting IFNγ-mediated T cell activity, which is at least partly modulated by HMGB1 activity.

Emulsified Isoflurane Protects Against Transient Focal Cerebral Ischemia Injury in Rats via the PI3K/Akt Signaling Pathway.

BACKGROUND: Phosphoinositide-3-kinase (PI3K)/protein kinase B (Akt) pathway activation may promote neuronal survival via neuroprotection during inflammation after cerebral ischemia. In this study, we investigated whether IV pretreatment with emulsified isoflurane (EI) could decrease ischemic brain injury related to the PI3K/Akt pathway. METHODS: Male Sprague-Dawley rats received different doses of IV EI (1, 2, 4, or 8 mL/kg/h) or Intralipid (8 mL/kg/h) for 30 minutes (n = 6-12 per group), followed by middle cerebral artery occlusion (MCAO) for 100 minutes to induce transient focal ischemia. The neurologic score and infarct volume were measured 48 hours after MCAO. Immunostaining, Western blot analysis, and an enzyme-linked immunosorbent assay were used to assess EI effects on the cell inflammatory response, high-mobility group box-1 release, and phosphorylated Akt (expression. LY294002, a PI3K inhibitor, was also infused into the ventricular space before EI to determine the effect of EI. RESULTS: Four milliliters per kilogram per hour of EI reduced the infarct size (21.08 ± 11.24 vs 37.09 ± 10.46, P = 0.006), improved neurologic scores after MCAO (1.13 ± 0.48 vs 1.95 ± 0.65, P = 0.015), significantly reinforced neuronal survival (982.7 ± 364.4 vs 439.8 ± 278.4, P = 0.036), and inhibited CD68 macrophage/macroglial infiltration in the ischemic core (188.2 ± 49.1 vs 282 ± 49.4, P = 0.018) compared with the vehicle group. In the EI pretreatment group, the serum high-mobility group box-1 concentration (3.62 ± 0.72 vs 5.73 ± 0.65, P < 0.001) was decreased, and the cerebral phosphorylated Akt level (50.33 ± 4.73 vs 37.5 ± 3.11, P = 0.007) was increased at 48 hours, which was inhibited by LY294002 compared with the vehicle group (5.31 ± 0.72 vs 5.73 ± 0.65, P = 0.216; 43.00 ± 4.84 vs 37.5 ± 3.11, P = 0.091). CONCLUSIONS: These findings suggest that EI pretreatment protects against ischemic brain injury via the inhibition of cerebral inflammation and is associated with the PI3K-Akt pathway in rats with MCAO. This drug may be a novel therapeutic agent for patients after stroke.