The first ER-targeting flavone-based fluorescent probe for Cys: Applications in real-time tracking in an epilepsy model and food analysis.

Epilepsy is one of the most commonly-seen neurological disorders, and both endoplasmic reticulum stress (ERS) and oxidative stress (OS) have been demonstrated to be associated with epileptic seizures. As one of the three endogenous thiol-containing amino acids, cysteine (Cys) is recognized not only as an important biomarker of various biological processes but also widely used as a significant additive in the food industry. However, the exact role that Cys plays in ERS has not been well answered up to now. In this paper, we reported the first flavone-based fluorescent probe (namely BFC) with nice endoplasmic reticulum (ER)-targeting ability, which was capable of monitoring Cys in a fast response (3.0 min), large stokes shift (130 nm) and low detection limit (10.4 nM). The recognition mechanism of Cys could be attributed to the addition-cyclization reaction involving a Cys residue and an acrylate group, resulting in the release of the strong excited-state intramolecular proton transfer (ESIPT) emission molecule of benzoflavonol (BF). The low cytotoxicity and good biocompatibility of the probe BFC allowed for monitoring the fluctuation of endogenous Cys levels under both ERS and OS processes, as well as in zebrafish models of epilepsy. Quantitative determination of Cys with the probe BFC was also achieved in three different food samples. Additionally, a probe-immersed test strips integrated with a smartphone device was successfully constructed for on-site colorimetric detection of Cys. Undoubtedly, our work provided a valuable tool for tracking Cys levels in both an epilepsy model and real food samples.

Shortening the early diagnostic window of Hg2+-induced liver injury with a H2O2-activated fluorescence/afterglow imaging assay.

Mercury ions (Hg2+) and mercury derivatives are a serious threat to ecosystems and human health due to their toxicity, and their toxicological effects are associated with a burst of reactive oxygen species (ROS) due to the oxidative stress. Endogenous hydrogen peroxide (H2O2), a featured ROS in vivo, plays an irreplaceable role in a significant number of pathological processes. However, the exact bioeffect role that H2O2 plays in Hg2+-induced oxidative stress in a specific disease has not been well answered. In particular, optical imaging probes for H2O2 endowed with afterglow emission properties are very rare. Here, the first fluorescence/afterglow probe (FA-H2O2) for accurate and specific detection of H2O2 in cells, zebrafish, and mice under Hg2+-induced oxidative stress is reported. Moreover, FA-H2O2 in its afterglow emission enables efficient monitoring of endogenous H2O2 with a higher signal-to-noise ratio (SNR) in comparison to its fluorescence signals. More importantly, by virtue of the merits of afterglow emission that can eliminate autofluorescence, thus for the first time, shortening the diagnostic window of Hg2+-induced liver injury with FA-H2O2 via noninvasive afterglow emission tracking of H2O2 is achieved, which definitely provides a new opportunity and promising tool for early diagnosis of Hg2+-induced liver injury.

UHPLC-Q/Orbitrap HRMS combined with spectrum-effect relationship and network pharmacology to discovery the gastrointestinal motility-promoting material basis in Citri Sarcodactylis Fructus.

ETHNOPHARMACOLOGICAL RELEVANCE: The prevalence of gastrointestinal motility disorders (GMD) is increasing and is characterized by long-term recurrence. Citri Sarcodactylis Fructus (CSF), as a traditional Chinese medicine (TCM) known in "regulating qi and harmonizing the stomach", has therapeutic effects on GMD. However, the material basis of its efficacy is not clear. AIM OF THE STUDY: The aim of this study was to evaluate the gastrointestinal motility-promoting activity of CSF extracts and to screen their active ingredients and to perform a preliminary validation. METHODS: The chemical composition spectrum of different extracts of CSF were established by ultra high-performance liquid chromatography coupled with quadrupole orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS). The gastrointestinal motility-promoting activities of CSF were investigated by determining the intestinal propulsion rate, gastric emptying rate, acetylcholinesterase activity, and motilin content in L-arginine-induced GMD mice. Spectrum-effect relationship and network pharmacology analysis were used for the screening of potential active ingredients. A zebrafish gastrointestinal motility model traced with Nile Red was established to validate the active ingredients. Molecular docking prediction was used to explore the mechanism of action of the active ingredient. Finally, Western blotting and TUNEL staining were performed to validate the molecular docking predictions. RESULTS: In total, 42 shared components were identified. The main active fraction of CSF to promote gastrointestinal motility was 70% ethanol elution fraction. Eleven potential active ingredients were screened by grey correlation analysis, orthogonal partial least squares analysis, and "active ingredient-target" network. Six compounds were confirmed as the pharmacodynamic substances of CSF by zebrafish gastrointestinal motility model, namely, quercetin, kaempferol, isorhamnetin, diosmetin, hesperetin, and 5,7,3'-trihydroxy-6,4',5'-trimethoxyflavone. Molecular docking predictions and Western blotting assays indicated that CSF may act on AKT and MMP9 targets to exert gastrointestinal motility-promoting activity. CONCLUSION: This study provided a foundation for elucidating the gastrointestinal motility-promoting activity of CSF and its material basis by integrating spectrum-effect relationship and network pharmacology. It also provided a theoretical basis for quality control of CSF and a new idea for the discovery and validation of pharmacodynamic substances in TCM.

Natural harmaline acts as novel fluorescent probe for hypochlorous acid and promising therapeutic candidate for rheumatoid arthritis.

Endogenous hypochlorous acid (HOCl) is one of the most important reactive oxygen species (ROS) and acts as a distinct biomarker that is involved in various inflammatory responses including rheumatoid arthritis (RA). Therefore, it's crucial to develop an efficient method for the tracking and analysis of HOCl levels in vivo. Natural products continue to be compounds of interest, because they not only offer diverse and specific molecular scaffolds but also provide invaluable sources for new drug discovery. Herein, we firstly demonstrated harmaline (HML), a natural alkaloid mainly found in Peganum harmala L, could be acted as a novel fluorescent probe for HOCl with exceptional precision and responsiveness. Remarkably, this probe not only specifically tracked HOCl levels in cells and inflammatory RA mouse models, but also exhibited effective anti-inflammatory effects on RAW264.7 cells and anti-proliferative effects on fibroblast-like synoviocytes. Furthermore, HML has the potential to alleviate LPS-induced inflammation by inhibiting the NF-κB signaling pathway. This study represents the first example of a natural product that can simultaneously act as a fluorescent probe for specific ROS and a promising therapeutic candidate for a specific disease, which will undoubtedly extend the application of fluorophore-rich natural products.

A novel AIE-based mitochondria-targeting fluorescent probe for monitoring of the fluctuation of endogenous hypochlorous acid in ferroptosis models.

Ferroptosis is a way of cell death mainly due to the imbalance between the production and degradation of lipid reactive oxygen species, which is closely associated with various diseases. Endogenous hypochlorous acid (HOCl) mainly produced in mitochondria is regarded as an important signal molecule of ferroptosis. Therefore, monitoring the fluctuation of endogenous HOCl is beneficial to better understand and treat ferroptosis-related diseases. Inspired by the promising aggregation-induced emission (AIE) properties of tetraphenylethene (TPE), herein, we rationally constructed a novel AIE-based fluorescent probe, namely QTrPEP, for HOCl with nice mitochondria-targeting ability and high sensitivity and selectivity. Probe QTrPEP consisted of phenylborate ester and the AIE fluorophore of quinoline-conjugated triphenylethylene (QTrPE). HOCl can brighten the strong fluorescence through a specific HOCl-triggered cleavage of the phenylborate ester bond and release of QTrPE, which has been demonstrated by MS, HPLC, and DLS experiments. In addition, combining QTrPE-doped test strips with a smartphone-based measurement demonstrated the excellent performance of the probe to sense HOCl. The obtained favorable optical properties and negligible cytotoxicity allowed the use of this probe for tracking of HOCl in three different cells. In particular, this work represents the first AIE-based mitochondria-targeting fluorescent probe for monitoring the fluctuation of HOCl in ferroptosis.

Astrocyte-TREM2 alleviates brain injury by regulating reactive astrocyte states following ischemic stroke.

Triggering receptor expressed on myeloid cells 2 (TREM2) has been shown to confer strong neuroprotective effects in acute ischemic stroke (AIS). However, as the vast majority of research findings to date are based on its functions in microglia, the precise role of TREM2 in astrocytes after AIS is unknown. Here, both loss- and gain-of-function experiments were employed to investigate how astrocytic TREM2 influences the pathogenesis of AIS in vivo and in vitro. Our results demonstrated that cerebral ischemia triggered induction of TREM2 expression on reactive astrocytes following AIS. In addition, astrocyte-specific TREM2 knockout mice exhibited much greater brain injury than TREM2 flox/flox controls following AIS, as evidenced by increased cerebral infarct volume, neuronal apoptosis and neurological deficit, which was associated with an increased expression of pro-inflammatory molecule complement component 3 (C3) on reactive astrocytes and activation of microglia/macrophages but decreased expression of S100 calcium binding protein A10 (S100A10) and arginase1 (Arg1) on reactive astrocytes. Mechanistic analyses revealed that astrocytic TREM2 alleviated brain injury by inhibiting detrimental actions of reactive astrocytes but promoting their neuro- and glioprotective actions via the kruppel-like transcription factor-4-nuclear factor-κB axis. Together, this study provides novel evidence for a critical protective role of astrocyte-derived TREM2 in AIS and highlights a potential therapeutic target for the treatment of AIS.

A novel LD-targeting cysteine-activated fluorescent probe for diagnosis of APAP-induced liver injury and its application in food analysis.

Cysteine (Cys) not only plays an indispensable role in maintaining the redox balance in organisms, but is also an important nutrient in the food industry. Fluorescence-based detection systems have emerged as an effective method to track the locations and concentrations of different species. To achieve efficient monitoring of Cys in both food samples and biological systems, a novel lipid droplet (LD) targeted fluorescent probe (namely NIT-Cys) was constructed for the turn-on detection of Cys, characterized by a large Stokes shift (142 nm), a short response time (<8 min), and a low Cys detection limit (39 nM). Furthermore, the NIT-Cys probe has been successfully used not only to quantify the amounts of Cys in selected food samples, but also to enable the visualization of endogenous Cys in acetaminophen (APAP)-induced drug-induced liver injury cells, zebrafish larvae and mice models. Consequently, the work presented here provides an efficient tool for monitoring Cys.

Real-time monitoring of endogenous cysteine in LPS-induced oxidative stress process with a novel lysosome-targeted fluorescent probe.

Cysteine (Cys), one of essential small-molecule-based biothiols in the human body, contributes to the regulation of redox reactions and is closely associated with many physiological and pathological metabolic processes. Herein, a novel fluorescent probe, hydroxyphenyl-conjugated benzothiazole (HBT-Cys) capable of detecting Cys was constructed, where acrylate served as the recognition group and hydroxyphenyl-linked benzothiazole acted as the fluorophore. The fluorescence of the probe was negligible in the absence of Cys, and an intense blue fluorescence was observed upon addition of Cys. The Cys-sensing mechanism could be ascribed to the Cys-involved hydrolysis reaction with acrylate, leading to light up the emission at 430 nm with about 80-fold enhancement. In addition, HBT-Cys exhibited a fast response time, remarkable selectivity and low detection limit. HBT-Cys also worked well in real-time monitoring of Cys in three different food samples (wolfberry, hawthorn, and red dates). Importantly, our probe had an excellent lysosomes-targeted ability, which was successfully employed to real-time visualize the fluctuation of both exogenous and endogenous Cys in living cells and zebrafish under lipopolysaccharide (LPS)-induced oxidative stress. Hopefully, the work shown here provides a potent candidate for the real-time tracking of Cys fluctuations in various biological samples.

Characterization of Global Research Trends and Prospects on Moyamoya Disease: Bibliometric Analysis.

BACKGROUND: Moyamoya disease (MMD) is a rare cerebrovascular disease in neurology. This study investigates the literature related to MMD from its discovery to the present and identifies research levels, achievements, and trends. METHODS: All publications on MMD from its discovery to present were downloaded from the Web of Science Core Collection on September 15, 2022 and bibliometric analyses were visualized by HistCite Pro, VOSviewer, Scimago Graphica, CiteSpace, and R language. RESULTS: There were 3414 articles in 680 journals by 10,522 authors in 2441 institutions and 74 countries/regions worldwise are included in the analyses. Since the discovery of MMD, output of publications has shown an upward trend. Japan, the United States, China, and South Korea are 4 major countries in MMD. The United States has the strongest cooperation with other countries. China's Capital Medical University is the output-leading institution worldwide, followed by Seoul National University and Tohoku University. The 3 authors with the most published articles are Kiyohiro Houkin, Dong Zhang, and Satoshi Kuroda. World Neurosurgery, Neurosurgery, and Stroke are the most recognized journals for researchers. Hemorrhagic moyamoya disease, susceptibility gene, and arterial spin are the primary focus areas of MMD research. "Rnf213,""vascular disorder," and "progress" are the top keywords. CONCLUSIONS: We analyzed publications of global scientific research on MMD systematically by bibliometric methods. This study can provide one of the most comprehensive and accurate analyses for MMD scholars worldwide.

The critical role of KLF4 in regulating the activation of A1/A2 reactive astrocytes following ischemic stroke.

BACKGROUND: We have previously demonstrated that the expression of kruppel-like transcription factor-4 (KLF-4) is upregulated in astrocytes following acute ischemic stroke (AIS) and found that KLF4 confers vascular protection against cerebral ischemic injury. However, the functional role of KLF4 in astrocyte after AIS is far from clear. METHODS: The intrinsic relationship between KLF4 and A1/A2 reactive astrocytes and the impact of astrocytic KLF4 on the activation of A1/A2 subtype astrocytes were evaluated in middle cerebral artery occlusion (MCAO) mice and oxygen-glucose deprivation and restoration (OGD/R) astrocytes. RESULTS: Our results demonstrated that astrocytic KLF4 expression and complement C3-positive A1 and S100 calcium binding protein A10 (S100A10)-positive A2 astrocytes were induced in the ischemic penumbra following focal cerebral ischemia, and the time course of upregulation of astrocytic KLF4 correlated closely with the activation of A2 astrocytes. The dual immunofluorescent studies displayed that in the ischemic hemisphere, where the high levels of KLF4 were expressed, there were relatively low levels of C3 expressed in the reactive astrocytes and vice versa, but KLF4 was always co-stained well with S100A10. Mechanistic analyses revealed that astrocytic KLF4 inhibited the activation of A1 astrocyte but promoted A2 astrocyte polarization after OGD/R by modulating expressions of nuclear factor-kB. CONCLUSIONS: Astrocyte-derived KLF4 has a critical role in regulating the activation of A1/A2 reactive astrocytes following AIS.

Serum inflammatory cell adhesion molecules predict malignant cerebral edema and clinical outcome early after mechanical thrombectomy in stroke.

BACKGROUND: Early prediction of life-threatening malignant cerebral edema (MCE) after mechanical thrombectomy (MT) is of clinical importance. Although inflammatory cell adhesion molecules (CAMs) and anti-inflammation factor Kruppel-like transcription factor (KLF) 4 are induced after acute ischemic stroke (AIS), the relationship between expressions of these molecules after MT and MCE as well as outcome in AIS patients have rarely been explored. METHODS: We retrospectively reviewed the data of all AIS patients with large-vessel occlusion in anterior circulation who underwent MT from our stroke centers. The serum levels of CAMs and KLF4 were determined at 12 h after MT. MCE was assessed on follow-up head computed tomography within 5 days after MT. RESULTS: Of 91 included patients, 18 (19.8 %) patients experienced MCE. Patients with MCE were more likely to have higher levels of E-selectin and inter-cellular adhesion molecule 1 (ICAM-1) than those without MCE (P < 0.05). More specifically, elevated E-selectin, but not of vascular cell adhesion molecule 1 (VCAM-1), ICAM-1 and KLF4, was significantly associated with MCE after adjusting for hypertension, admission NIHSS, Alberta Stroke Program Early CT Scores, serum glucose, collateral circulation and onset to recanalization time respectively (P < 0.05). ROC curve suggested that E-selectin had considerable discrimination to predict MCE (AUC=0.7, 95 % CI: 0.55-0.83). Moreover, after adjusting by confounders, serum levels of E-selectin and ICAM-1 were independently associated with 3-month outcome in AIS patients after MT (both P < 0.05). CONCLUSIONS: These data indicate that of three CAMs, serum E-selectin level early after MT is the best predictor for MCE and outcome in AIS.

Naoxintong Capsule for Secondary Prevention of Ischemic Stroke: A Multicenter, Randomized, and Placebo-Controlled Trial.

OBJECTIVE: To examine whether the combination of Naoxintong Capsule with standard care could further reduce the recurrence of ischemic stroke without increasing the risk of severe bleeding. METHODS: A total of 23 Chinese medical centers participated in this trial. Adult patients with a history of ischemic stroke were randomly assigned in a 1:1 ratio using a block design to receive either Naoxintong Capsule (1.2 g orally, twice a day) or placebo in addition to standard care. The primary endpoint was recurrence of ischemic stroke within 2 years. Secondary outcomes included myocardial infarction, death due to recurrent ischemic stroke, and all-cause mortality. The safety of drugs was monitored. Results were analyzed using the intention-to-treat principle. RESULTS: A total of 2,200 patients were enrolled from March 2015 to March 2016, of whom 143 and 158 in the Naoxintong and placebo groups were lost to follow-up, respectively. Compared with the placebo group, the recurrence rate of ischemic stroke within 2 years was significantly lower in the Naoxintong group [6.5% vs. 9.5%, hazard ratio (HR): 0.665, 95% confidence interval (CI): 0.492-0.899, P=0.008]. The two groups showed no significant differences in the secondary outcomes and safety, including rates of severe hemorrhage, cerebral hemorrhage and subarachnoid hemorrhage (P>0.05). CONCLUSION: The combination of Naoxintong Capsule with standard care reduced the 2-year stroke recurrence rate in patients with ischemic stroke without increasing the risk of severe hemorrhage in high-risk patients. (Trial registration No. NCT02334969).

Interactions Between Meropenem and Renal Drug Transporters.

BACKGROUND: Meropenem is a carbapenem antibiotic and is commonly used with other antibiotics for the treatment of bacterial infections. It is primarily eliminated renally by glomerular filtration and renal tubular secretion. OBJECTIVE: This study aimed to evaluate the roles of renal uptake and efflux transporters in the excretion of meropenem and potential drug interactions mediated by renal drug transporters. METHODS: Uptake and inhibition studies were conducted in human embryonic kidney 293 cells stably transfected with Organic Anion Transporter (OAT) 1, OAT3, Multidrug and Toxin Extrusion Protein (MATE) 1, and MATE2K, as well as membrane vesicles containing breast cancer resistance-related protein (BCRP), multidrug resistance protein 1 (MDR1), and Multidrug Resistance-associated Protein 2 (MRP2). Probenecid and piperacillin were used to assess potential drug interactions with meropenem in rats. RESULTS: We observed that meropenem was a low-affinity substrate of OAT1/3 and had a weak inhibitory effect on OAT1/3 and MATE2K. BCRP, MDR1, MRP2, MATE1, and MATE2K could not mediate renal excretion of meropenem. Moreover, meropenem was not an inhibitor of BCRP, MDR1, MRP2, or MATE1. Among five tested antibiotics, moderate inhibition on OAT3-mediated meropenem uptake was observed for linezolid (IC50 value was 69.2 µM), weak inhibition was observed for piperacillin, benzylpenicillin, and tazobactam (IC50 values were 282.2, 308.0 and 668.1 µM, respectively), and no inhibition was observed for sulbactam. Although piperacillin had a relatively high drug-drug interaction index (ratio of maximal unbound plasma concentration to IC50 was 1.42) in vitro, no meaningful impact was reported on the pharmacokinetics of meropenem in rats. CONCLUSION: Our results indicated that clinically significant interactions between meropenem and these five antibiotics are low.

[Effects of water level variations on survival, morphological phenotype and responsiveness of tadpoles to phytohemagglutinin]. / æ°´ä½å˜åŒ–å½±å“èŒèšªå­˜æ´»ã€å½¢æ€è¡¨åž‹å’Œæ¤ç‰©è¡€å‡ç´ ååº”.

To investigate the responses of morphological and physiological characteristics in tadpoles of Pelophylax nigromaculatus to various water levels, a total of 480 tadpoles with similar sizes were treated with rapid, moderate and slow desiccation. Both the metamorphosis duration and average metamorphosis survival rate of specific developmental stages were counted. Their body and internal organ sizes, as well as the responsiveness to phytohemagglutinin (PHA-P) of froglets were measured after complete metamorphosis. The results showed that the metamorphosis duration of tadpoles in the control group was the longest. The average metamorphosis survival rate of different treatment groups ranged from 72.5% to 90.8%, which was the highest in the control group and the lowest in the rapid desiccation group. Body weight (BW) and body length (BL) of froglets in control and slow desiccation groups were significantly higher than those of moderate or rapid desiccation groups. Body width, ratio of BW to BL, carcass wet weight and the wet weight index of lung and fat body were the lowest in rapid desiccation group. There was no significant difference in the wet weight index of heart, spleen, kidneys and digestive organs, as well as the length index of different segments of whole digestive tract between groups. The maximal response value of froglets to PHA-P appeared at 3 h after injection in different treatments, which was significantly higher in moderate and slow desiccation groups than the control group. There was no significant difference between rapid desiccation group and any other groups. The tadpoles of P. nigromaculatus might accelerate the metamorphosis developmental process to cope with desiccation stress, accompanying with decreased atrophic body size and weakened cell-mediated immunity, which would harm tadpoles to successfully land.

Silencing of lncRNA XIST impairs angiogenesis and exacerbates cerebral vascular injury after ischemic stroke.

The aim of this study was to investigate the function and regulatory mechanism of long non-coding RNA (lncRNA) X-inactive-specific transcript (XIST) in cerebral ischemic stroke (CIS). The impact of lncRNA XIST on CIS was evaluated in acute CIS patients, middle cerebral artery occlusion (MCAO) mice, and oxygen-glucose deprivation and restoration brain endothelial cells. Our results demonstrated that the expression of lncRNA XIST decreased during the early stages of CIS but then increased in the later stages in CIS patients and ischemic models in vivo and in vitro. In addition, the serum levels of lncRNA XIST negatively correlated with severity of neurological impairment of CIS patients. Further studies exhibited that lncRNA XIST regulated the expression of proangiogenic factor-integrin α5 (Itgα5) and anti-inflammation factor-Kruppel-like transcription factor 4 (KLF4) by targeting microRNA-92a (miR-92a). Silencing of lncRNA XIST impaired angiogenesis and exacerbated cerebral vascular injury following CIS, leading to larger infarcts and worse neurological deficits in transient MCAO mice. Mechanistic analysis revealed that lncRNA XIST modulated angiogenesis and alleviated cerebral vascular injury following CIS through mediating the miR-92a/Itgα5 or KLF4 axis, respectively. These data indicate that lncRNA XIST confers protection against CIS, providing a valuable target for future prevention and treatment of CIS.

Upregulation of circ-FBL promotes myogenic proliferation in myasthenia gravis by regulation of miR-133/PAX7.

Myasthenia gravis (MG) is a disease involving neuromuscular transmission that causes fatigue of skeletal muscles and fluctuating weakness. It has been shown that impairment of myogenic differentiation and myofiber maturation may be the underlying cause of MG. In this study, we detected the abnormal expression of circular RNA (circRNA) using next-generation sequencing in patients with MG. We then investigated the regulatory mechanism and the relationship among circRNA, microRNA, and messenger RNA using quantitative reverse-transcription polymerase chain reaction, bioinformatics analysis, and luciferase report analysis. The expression of inflammatory cytokines and regulatory T lymphocytes was shown to be increased. Circ-FBL was significantly increased in MG patients. Bioinformatics and luciferase report analyses confirmed that miR-133 and PAX7 were the downstream targets of circ-FBL. Overexpression of circ-FBL promoted myoblast proliferation by regulation of miR-133/PAX7. Taken together, our study showed that upregulation of circ-FBL promoted myogenic proliferation in patients with MG by regulating miR-133/PAX7.

Characteristics and prognosis of acute basilar artery occlusion in minor to moderate stroke and severe stroke after endovascular treatment: A multicenter retrospective study.

OBJECTIVE: We aimed to investigate characteristics and outcomes of patients receiving mechanical thrombectomy (MT) between minor to moderate stroke and severe stroke caused by acute basilar artery occlusion (BAO). METHODS: We retrospectively reviewed the data of all patients with BAO who underwent MT from three stroke centers between January 2016 and January 2020. The patients were dichotomized as minor to moderate or severe stroke group according to their admission National Institutes of Health Stroke Scale (NIHSS) score <21and ≥21. Patient characteristics, imaging findings, and outcomes were compared between the two groups. RESULTS: A total of 72 patients were included in this study. The posterior circulation Alberta Stroke Program Early CT Score (PC-ASPECTS) in the minor to moderate stroke patients were significantly higher than that of patients with severe stroke (P = 0.013). The good posterior circulation collateral scores (PC-CS) (6-10) were more commonly found in patients with minor to moderate stroke than in patients with severe stroke (58.14 % vs 10.34 %,P < 0.001). There were similar rates of successful recanalization between the two groups. Patients with minor to moderate stroke had a higher rate of favorable outcomes (mRS score 0-2, 60.47 % vs 20.69 %, P = 0.002) and a lower rate of periprocedural complications (4.65 % vs 31.03 %, P = 0.005) and mortality (4.65 % vs 24.14 %, P = 0.026) at 3 months after MT compared with the patients with severe stroke. CONCLUSIONS: Acute BAO patients with minor to moderate stroke had better posterior circulation collateral and had better outcomes after MT than those patients with severe stroke.

Baicalin and the liver-gut system: Pharmacological bases explaining its therapeutic effects.

With the development of high-throughput screening and bioinformatics technology, natural products with a range of pharmacological targets in multiple diseases have become important sources of new drug discovery. These compounds are derived from various plants, including the dried root of Scutellaria baicalensis Georgi, which is often used as a traditional Chinese herb named Huangqin, a popular medication used for thousands of years in China. Many studies have shown that baicalin, an extract from Scutellaria baicalensis Georgi, exerts various protective effects on liver and gut diseases. Baicalin plays a therapeutic role mainly by mediating downstream apoptosis and immune response pathways induced by upstream oxidative stress and inflammation. During oxidative stress regulation, PI3K/Akt/NRF2, Keap-1, NF-κB and HO-1 are key factors associated with the healing effects of baicalin on NAFLD/NASH, ulcerative colitis and cholestasis. In the inflammatory response, IL-6, IL-1ß, TNF-α, MIP-2 and MIP-1α are involved in the alleviation of NAFLD/NASH, cholestasis and liver fibrosis by baicalin, as are TGF-ß1/Smads, STAT3 and NF-κB. Regarding the apoptosis pathway, Bax, Bcl-2, Caspase-3 and Caspase-9 are key factors related to the suppression of hepatocellular carcinoma and attenuation of liver injury and colorectal cancer. In addition to immune regulation, PD-1/PDL-1 and TLR4-NF-κB are correlated with the alleviation of hepatocellular carcinoma, ulcerative colitis and colorectal cancer by baicalin. Moreover, baicalin regulates intestinal flora by promoting the production of SCFAs. Furthermore, BA is involved in the interactions of the liver-gut axis by regulating TGR5, FXR, bile acids and the microbiota. In general, a comprehensive analysis of this natural compound was conducted to determine the mechanism by which it regulates bile acid metabolism, the intestinal flora and related signaling pathways, providing new insights into the pharmacological effects of baicalin. The mechanism linking the liver and gut systems needs to be elucidated to draw attention to its great clinical importance.

The AMPK-MFN2 axis regulates MAM dynamics and autophagy induced by energy stresses.

Energy deprivation activates the cellular energy sensor AMP-activated protein kinase (AMPK), which in turn induces macroautophagy/autophagy. The mitochondrial-associated ER membrane (MAM) plays a key role in mitochondrial division and autophagy, and the mitochondrial fusion protein MFN2 (mitofusin 2) tethers the MAM, but the mechanism by which AMPK and MFN2 regulate autophagy in response to energy stress remains unclear. Here, we found that energy stress not only triggers mitochondrial fission and autophagy, but more importantly increases the number of MAMs, a process that requires AMPK. Interestingly, under energy stress, considerable amounts of AMPK translocate from cytosol to the MAM and the mitochondrion as mitochondrial fission occurs. Unexpectedly, AMPK interacts directly with MFN2. The autophagic ability of mouse embryonic fibroblasts (MEFs) lacking MFN2 (mfn2-/-) is significantly attenuated in response to energy stress as compared to wild-type MEFs (WT MEFs), while re-expression of MFN2 in mfn2-/- cells rescues the autophagy defects of these cells. The abundance of MAMs is also greatly reduced in MFN2-deficient cells. Functional experiments show that the oxygen consumption rate and the glycolytic function of cells lacking MFN2 but not MFN1 are obviously attenuated, and MFN2 is important for cell survival under energy stress. In conclusion, our study establishes the molecular link between the energy sensor AMPK and the MAM tether MFN2, and reveals the important role of AMPK and MFN2 in energy stress-induced autophagy and MAM dynamics.Abbreviations: ACTB, actin beta; AMPK, AMP-activated protein kinase; BECN1, beclin 1; CANX, calnexin; ER, endoplasmic reticulum; HRP, horseradish peroxidase; EM, electron microscopy; FL, full-length; KD, kinase dead, KO, knockout; MAb, monoclonal antibody; MAMs, mitochondria-associated membranes; MAP1LC3/LC3B, microtubule associated protein 1 light chain 3; MFN2, mitofusin 2; OPA1, OPA1 mitochondrial dynamin like GTPase; PAb, polyclonal antibody; PtdIns3K, class III phosphatidylinositol 3-kinase; PtdIns3P, phosphatidylinositol 3-phosphate; SD, standard deviation; TEM, transmission electron microscopy; TOMM20, translocase of outer mitochondrial membrane 20; ULK1, unc-51 like autophagy activating kinase 1; MEF, mouse embryonic fibroblast; WT, wildtype.