Acute Stage Of Ischemic Stroke Research Articles

The underlying mechanism of calcium toxicity-induced autophagic cell death and lysosomal degradation in early stage of cerebral ischemia.

Cerebral ischemia is the important cause of worldwide disability and mortality, that is one of the obstruction of blood vessels supplying to the brain. In early stage, glutamate excitotoxicity and high level of intracellular calcium (Ca2+) are the major processes which can promote many downstream signaling involving in neuronal death and brain tissue damaging. Moreover, autophagy, the reusing of damaged cell organelles, is affected in early ischemia. Under ischemic conditions, autophagy plays an important role to maintain energy of the brain and its function. In the other hand, over intracellular Ca2+ accumulation triggers excessive autophagic process and lysosomal degradation leading to autophagic process impairment which finally induce neuronal death. This article reviews the association between intracellular Ca2+ and autophagic process in acute stage of ischemic stroke.

Low-intensity focused ultrasound stimulation promotes stroke recovery via astrocytic HMGB1 and CAMK2N1 in mice

BackgroundLow-intensity focused ultrasound stimulation (LIFUS) has been developed to enhance neurological repair and remodelling during the late acute stage of ischaemic stroke in rodents. However, the cellular and molecular mechanisms...

Pulse pressure level after acute ischemic stroke is associated with early neurological deterioration.

Early neurological deterioration (END) is an unfavorable outcome of acute ischemic stroke and is associated with poor prognosis. Blood pressure variability has been suggested to be involved in the development of END. Therefore, the present study investigated the association between blood pressure variability and the development of END. In the present prospective observational study, 286 patients who developed acute ischemic stroke and then hospitalized within 24 h of stroke onset were recruited. Blood pressure parameters (systolic blood pressure, diastolic blood pressure and pulse pressure) were monitored using a 24 h ambulatory sphygmomanometer within 72 h of ischemic onset. Clinical characteristics were also recorded. Multivariate logistic regression analysis was used to analyze the possible relationship between blood pressure parameters and END after adjustment for confounders. Of the 286 patients in the present study, 64 (22.3%) developed END. Pulse pressure variables, including the mean of 24-h pulse pressure (24-h PPMEAN) and the mean of daytime pulse pressure (Day PPMEAN), were found to be higher in the END group compared with those in the non-END group (P<0.05). Multivariate logistic regression analysis revealed that the blood pressure parameters 24-h PPMEAN [odds ratio (OR), 1.08; 95% CI, 1.01-1.16; P=0.02) and Day PPMEAN (OR, 1.20; 95% CI, 1.011-1.45; P=0.04) were significantly associated with END. These findings suggest that the pulse pressure level fluctuations during the acute stage of ischemic stroke can serve important roles in the development of END, which worsens outcomes after stroke.

Increasing and high prevalence of moderate to severe obstructive sleep apnea in acute ischemic stroke in Taiwan

The prevalence of obstructive sleep apnea (OSA) after stroke is variable, likely due to the time of examination and patient population. Although risk factors for OSA are well established, those for OSA in patients with ischemic stroke have not yet been fully identified. Therefore, we examined the prevalence of OSA and identified risk factors for OSA in the acute stage of ischemic stroke in the Taiwanese population. A total of 103 patients with acute ischemic stroke were screened for OSA by performing polysomnography. The demographic and clinical data, Epworth Sleepiness Scale score, and other stroke risk factors were recorded. Sleep parameters, namely sleep efficiency, sleep stages, apnea-hypopnea index, and oxygen desaturation index were recorded. Logistic regression analysis was conducted to determine clinical and demographic risk factors for moderate to severe OSA in patients with stroke. We determined that 91.2% of the patients had OSA in the acute stage of ischemic stroke, and 70% of the patients had moderate to severe OSA. Multivariate logistic regression analysis revealed that patients aged ≥65 years had a significantly higher risk of moderate to severe OSA (adjusted OR: 3.04, 95% CI: 1.20-7.69, p<0.05) compared with patients with ischemic stroke aged <50 years. OSA is highly prevalent among patients with ischemic stroke in the acute stage, and those aged ≥65 years had a significantly increased risk of moderate to severe OSA. In clinical practice, routine PSG screening of OSA may be necessary among older patients with stroke.

Radiological Assessment of Aphasia Outcome: A Case Report

Background Stroke is a leading cause of major adult disability, and aphasia is one of the most common consequences of stroke. It has been reported that 21%–38% of patients with stroke have aphasia in the acute stage. Case presentation A 64- year old Egyptian male diagnosed with recent ischemic stroke in the form of the form of inability to express himself in spoken words; Broca’s aphasia. Diffusion tensor tractography (DTT) was done to assess the integrity of the arcuate fasciculus in the acute stage as a method to predict the possible outcome of the patient aphasia. DTT showed affection of broca’s territory of the arcuate fasciculus (AF) and it could be reconstructed measured by fractionated anisotropy (0.469). The patient initial aphasia test measured (47). Three months later, the patient improved and follow up aphasia test measured (74.4). Results Reconstruction of the arcuate fasciculus by DTT in the early stage of aphasia as shown in our patient carries a good clinical prognosis. Our patient aphasia improved markedly at 3 months follow up visit. Conclusion DTT could be a helpful tool to assess the outcome of aphasia at the early stage of acute ischemic stroke.

Scutellaria baicalensis Attenuated Neurological Impairment by Regulating Programmed Cell Death Pathway in Ischemic Stroke Mice.

Stroke is a major global health problem that causes significant mortality and long-term disability. Post-stroke neurological impairment is a complication that is often underestimated with the risk of persistent neurological deficits. Although traditional Chinese medicines have a long history of being used for stroke, their scientific efficacy remains unclear. Scutellaria baicalensis, an herbal component known for its anti-inflammatory and antioxidant properties, has traditionally been used to treat brain disorders. This study investigated the therapeutic effects of the Scutellaria baicalensis extraction (SB) during the acute stage of ischemic stroke using photothrombotic (PTB)-induced and transient middle cerebral artery occlusion (tMCAO) model mice. We found that SB mitigated ischemic brain injury, as evidenced by a significant reduction in the modified neurological severity score in the acute stage of PTB and both the acute and chronic stages of tMCAO. Furthermore, we elucidated the regulatory role of SB in the necroptosis and pyroptosis pathways during the acute stage of stroke, underscoring its protective effects. Behavioral assessments demonstrated the effectiveness of SB in ameliorating motor dysfunction and cognitive impairment compared to the group receiving the vehicle. Our findings highlight the potential of SB as a promising therapeutic candidate for stroke. SB was found to help modulate the programmed cell death pathways, promote neuroprotection, and facilitate functional recovery.

Pharmacological Upregulation of Microglial Lipid Droplet Alleviates Neuroinflammation and Acute Ischemic Brain Injury

Lipid droplets (LDs) were reported to play an important role in the modulation of inflammation and various cellular processes among multiple cell types. However, LDs accumulation, its function and mechanisms of its formation during ischemic stroke remained poorly-identified. In this study, we observed increased LDs accumulation in microglia at the acute stage of ischemic stroke by immunofluorescence and flow cytometry. Transcriptomic analysis indicated that microglia accumulated with LDs were associated with inflammation and phagocytosis. Both inflammatory activation and phagocytosis of tissue debris in microglia could contribute to LDs formation. Moreover, through specific LDs depletion and overload experiments by pharmacological approaches, we proposed that LDs was critical for the maintenance of anti-inflammatory properties of microglia. Furthermore, Atglistatin, a specific adipose triglyceride lipase (ATGL) inhibitor, was shown to prevent proinflammatory cytokines production in primary microglia through decreased LDs lipolysis. After Atglistatin treatment, middle cerebral artery occlusion (MCAO) mice showed decreased infarct volume and improved neurobehavioral performance at the acute stage of stroke. Our findings provided a biological basis for microglial LDs regulation as a potential therapeutic strategy for acute ischemic stroke and uncovered the neuroprotective role of Atglistatin in the treatment of MCAO mice.

Difference in acute and chronic stage ischemic stroke metabolic markers with controls

BackgroundAcute Ischemic Stroke (AIS), a major cause of disability, was previously associated with multiple metabolomic changes, but many findings were contradictory. Case-control and longitudinal study designs could have played a role in that. To clarify metabolomic changes, we performed a simultaneous comparison of ischemic stroke metabolome in acute, chronic stages of stroke and controls. MethodsThrough the nuclear magnetic resonance (NMR) platform, we evaluated 271 serum metabolites from a cohort of 297 AIS patients in acute and chronic stages and 159 controls. We used Sparse Partial Least Squares-Discriminant analysis (sPLS-DA) to evaluate group disparity; multivariate regression to compare metabolome in acute, chronic stages of stroke and controls; and mixed regression to compare metabolome acute and chronic stages of stroke. We applied false discovery rate (FDR) to our calculations. ResultsThe sPLS-DA revealed separation of the metabolome in acute, chronic stages of stroke and controls. Regression analysis identified 38 altered metabolites. Ketones, branched-chain amino acids (BCAAs), energy, and inflammatory compounds were mostly elevated, while alanine and glutamine were decreased in the acute stage. These metabolites declined/increased in the chronic stage, often to the same levels as in controls. Levels of fatty acids, phosphatidylcholines, phosphoglycerides, and sphingomyelins did not change between acute and chronic stages, but were different comparing to controls. ConclusionOur pilot study identified metabolites associated with acute stage of ischemic stroke and those that are altered in stroke patients comparing to controls regardless of stroke acuity. Future investigation in a larger independent cohort is needed to validate these findings.

U-shaped association between low-density lipoprotein cholesterol levels and risk of all-cause mortality mediated by post-stroke infection in acute ischemic stroke

During the acute stage of ischemic stroke, it remains unclear how to interpret the low low-density lipoprotein cholesterol (LDL-C) level. We aimed to evaluate the association between LDL-C levels, post-stroke infection, and all-cause mortality. 804,855 ischemic stroke patients were included. Associations between LDL-C levels, infection, and mortality risk were estimated by multivariate logistic regression models and displayed by restricted cubic spline curves. Mediation analysis was performed under counterfactual framework to elucidate the mediation effect of post-stroke infection. The association between LDL-C and mortality risk was U-shaped. The nadir in LDL-C level with the lowest mortality risk was 2.67 mmol/L. Compared with the group with LDL-C = 2.50–2.99 mmol/L, the multivariable-adjusted odds ratio for mortality was 2.22 (95% confidence intervals (CI): 1.77–2.79) for LDL-C <1.0 mmol/L and 1.22 (95% CI: 0.98–1.50) for LDL-C ≥5.0 mmol/L. The association between LDL-C and all-cause mortality was 38.20% (95% CI: 5.96–70.45, P = 0.020) mediated by infection. After stepwise excluding patients with increasing numbers of cardiovascular risk factors, the U-shaped association between LDL-C and all-cause mortality and the mediation effects of infection remained consistent with the primary analysis, but the LDL-C interval with the lowest mortality risk increased progressively. The mediation effects of infection were largely consistent with the primary analysis in subgroups of age ≥65 years, female, body mass index <25 kg/m2, and National Institutes of Health Stroke Scale ≥16. During the acute stage of ischemic stroke, there is a U-shaped association between LDL-C level and all-cause mortality, where post-stroke infection is an important mediating mechanism.

Cognitive impairment in the acute period of cardioembolic stroke

Background. Acute disorders of cerebral circulation are one of the most urgent problems of modern clinical neurology. Its significance is due to both the high frequency of occurrence and a large number of unsatisfactory outcomes, as well as the significant impact on the quality of life. Cognitive disorders are one of the main syndromes that manifest in post-stroke period and are associated with the rehabilitation potential, as well as opportunities for social and household adaptation. Our objective was to study the profile of cognitive impairment in a continuous sample of patients with cardioembolic stroke in the acute period of stroke, as well as to determine the correlations between their occurrence and the factors determining the course of the disease. Design and methods. The study involved 46 patients of both sexes in the acute stage of ischemic stroke (average age was 69 ± 11 years). The selection of patients was carried out in accordance with the diagnostic criteria of the International Association of Vascular Behavioral and Cognitive Disorders VASCOG. The results of the Hachinski scale were also considered. All patients underwent interview for history, complaints, as well as the assessment of the neurological status. Risk factors for stroke were evaluated. We also assessed the results of computer and magnetic resonance imaging, standard “stroke” scales and performed a neuropsychological examination aimed at a comprehensive study of cognitive functions. All studies were conducted at the end of 10-14 days from the development of focal neurological symptoms. Results. Cognitive disorders in patients with cardioembolic ischemic stroke turned out to be quite heterogeneous. Their severity in the majority of our patients was considered as mild cognitive impairment. Changes at dementia level were much less common and were characteristic of patients with extensive lesions or in the case of disorders in the field of cognitively significant “strategic” foci. Neurodynamic disorders predominated in the profile of disorders of higher cortical functions. Memory impairment was expressed slightly and was of a secondary nature. With the aggravation of cognitive disorders, mnestic disorders progressed. In addition, regulatory disorders began to be detected. The most significant correlations were found between the results of neurocognitive tests on the one hand and the assessment on the Bartel scales, the assessment of changes in white matter, as well as the volume of the necrosis focus, the presence of heart failure and dyscirculatory encephalopathy in the anamnesis. Conclusions. Cognitive impairment is a significant component of the clinical picture of cardioembolic stroke. Their severity is largely determined by the volume and topical localization of the ischemic focus, as well as the presence and severity of previous chronic cerebrovascular pathology. This determines the need to develop methods to assess the state of cognitive functions before the development of stroke.

Transcutaneous Electrical Nerve Stimulation (TENS) Alleviates Brain Ischemic Injury by Regulating Neuronal Oxidative Stress, Pyroptosis, and Mitophagy.

As a noninvasive treatment, transcutaneous electrical nerve stimulation (TENS) has been utilized to treat various diseases in clinic. However, whether TENS can be an effective intervention in the acute stage of ischemic stroke still remains unclear. In the present study, we aimed to explore whether TENS could alleviate brain infarct volume, reduce oxidative stress and neuronal pyroptosis, and activate mitophagy following ischemic stroke. TENS was performed at 24 h after middle cerebral artery occlusion/reperfusion (MCAO/R) in rats for 3 consecutive days. Neurological scores, the volume of infarction, and the activity of SOD, MDA, GSH, and GSH-px were measured. Moreover, western blot was performed to detect the related protein expression, including Bcl-2, Bax, TXNIP, GSDMD, caspase-1, NLRP3, BRCC3, HIF-1α, BNIP3, LC3, and P62. Real-time PCR was performed to detect NLRP3 expression. Immunofluorescence was performed to detect the levels of LC3. There was no significant difference of neurological deficit scores between the MCAO group and the TENS group at 2 h after MCAO/R operation (P > 0.05), while the neurological deficit scores of TENS group significantly decreased in comparison with MCAO group at 72 h following MACO/R injury (P < 0.05). Similarly, TENS treatment significantly reduced the brain infarct volume compared with the MCAO group (P < 0.05). Moreover, TENS decreased the expression of Bax, TXNIP, GSDMD, caspase-1, BRCC3, NLRP3, and P62 and the activity of MDA as well as increasing the level of Bcl-2, HIF-1α, BNIP3, and LC3 and the activity of SOD, GSH, and GSH-px (P < 0.05). In conclusion, our results indicated that TENS alleviated brain damage following ischemic stroke via inhibiting neuronal oxidative stress and pyroptosis and activating mitophagy, possibly via the regulation of TXNIP, BRCC3/NLRP3, and HIF-1α/BNIP3 pathways.

Loganin attenuates neuroinflammation after ischemic stroke and fracture by regulating α7nAChR-mediated microglial polarization.

Fracture in acute stage of ischemic stroke can increase inflammatory response and enhance stroke injury. Loganin alleviates the symptoms of many inflammatory diseases through its anti-inflammatory effect, but its role in ischemic stroke and fracture remains to be explored. Here, mice were handled with permanent middle cerebral artery occlusion (pMCAO) followed by tibial fracture 1 day later to establish a pMCAO+fracture model. Loganin or Methyllycaconitine (MLA, a specific a7nAchR inhibitor) were intragastrically administered 2 or 0.5h before pMCAO, respectively. And mouse motor function and infarct volume were evaluated 3 days after pMCAO. We found that loganin alleviated the neurological deficit, cerebral infarction volume, and neuronal apoptosis (NeuN+ TUNEL+ ) in mice with pMCAO+fracture. And loganin suppressed pMCAO+fracture-induced neuroinflammation by promoting M2 microglia polarization (Iba1+ CD206+ ) and inhibiting M1 microglia polarization (Iba1+ CD11b+ ). While administration with MLA reversed the protective effect of loganin on pMCAO+fracture-induced neurological deficit and neuroinflammation. Next, LPS was used to stimulate BV2 microglia to simulate pMCAO+fracture-induced inflammatory microenvironment in vitro. Loganin facilitated the transformation of LPS-stimulated BV2 cells from M1 pro-inflammatory state (CD11b+ ) to M2 anti-inflammatory state (CD206+ ), which was antagonized by treatment with MLA. And loganin induced autophagy activation in LPS-stimulated BV2 cells by activating a7nAchR. Moreover, treatment with rapamycin (an autophagy activator) neutralized the inhibitory effect of MLA on loganin induced transformation of BV2 cells to M2 phenotype. Furthermore, BV2 cells were treated with LPS, LPS + loganin, LPS + loganin+MLA, or LPS + loganin+MLA+ rapamycin to obtain conditioned medium (CM) for stimulating primary neurons. Loganin reduced the damage of primary neurons caused by LPS-stimulated BV2 microglia through activating a7nAchR and inducing autophagy activation. In conclusion, loganin played anti-inflammatory and neuroprotective roles in pMCAO + fracture mice by activating a7nAchR, enhancing autophagy and promoting M2 polarization of microglia.

Optogenetic Activation of Astrocytes Reduces Blood-Brain Barrier Disruption via IL-10 In Stroke.

Optogenetics has been used to regulate astrocyte activity and modulate neuronal function after brain injury. Activated astrocytes regulate blood-brain barrier functions and are thereby involved in brain repair. However, the effect and molecular mechanism of optogenetic-activated astrocytes on the change in barrier function in ischemic stroke remain obscure. In this study, adult male GFAP-ChR2-EYFP transgenic Sprague-Dawley rats were stimulated by optogenetics at 24, 36, 48, and 60 h after photothrombotic stroke to activate ipsilateral cortical astrocytes. The effects of activated astrocytes on barrier integrity and the underlying mechanisms were explored using immunostaining, western blotting, RT-qPCR, and shRNA interference. Neurobehavioral tests were performed to evaluate therapeutic efficacy. The results demonstrated that IgG leakage, gap formation of tight junction proteins, and matrix metallopeptidase 2 expression were reduced after optogenetic activation of astrocytes (p<0.05). Moreover, photo-stimulation of astrocytes protected neurons against apoptosis and improved neurobehavioral outcomes in stroke rats compared to controls (p<0.05). Notably, interleukin-10 expression in optogenetic-activated astrocytes significantly increased after ischemic stroke in rats. Inhibition of interleukin-10 in astrocytes compromised the protective effects of optogenetic-activated astrocytes (p<0.05). We found for the first time that interleukin-10 derived from optogenetic-activated astrocytes protected blood-brain barrier integrity by decreasing the activity of matrix metallopeptidase 2 and attenuated neuronal apoptosis, which provided a novel therapeutic approach and target in the acute stage of ischemic stroke.

Nomogram Prediction for Lower Extremity Deep Vein Thrombosis in Acute Ischemic Stroke Patients Receiving Thrombolytic Therapy.

This study developed and evaluated a tailored nomogram to predict the potential occurrence of early lower extremity deep vein thrombosis (LDVT) in patients receiving thrombolytic therapy. We performed several logistic analyses on the training cohort and created a corresponding nomogram to forecast early LDVT. The classification accuracy and the accuracy of predicted probabilities of the multiple logistic regression model were evaluated using area under the curve (AUC) and the calibration graph method. According to the multivariate logistic regression model homocysteine, previous history of hypertension and atrial fibrillation, indirect bilirubin, age, and sex was identified as independent determinants of early LDVT. The nomogram was constructed using these variables. The calibration plots showed a good agreement between the predicted and observed LDVT possibilities in the training and validation cohorts with AUCs being 0.833 (95% CI: 0.774-0.892) and 0.907 (95% CI: 0.801-1.000), respectively. Our nomogram offers clinicians a tool for predicting the individual risk of LDVT in the early stage of acute ischemic stroke in patients receiving thrombolytic therapy, which could lead to early intervention.

Microglia Exhibit Distinct Heterogeneity Rather than M1/M2 Polarization within the Early Stage of Acute Ischemic Stroke.

The classification of microglial M1/M2 polarization in the acute phase of ischemic stroke remains controversial, which has limited further advances in neuroprotective strategy. To thoroughly assess the microglial phenotypes, we made the middle cerebral artery occlusion model in mice to simulate the acute pathological processes of ischemic stroke from normal conditions to acute cerebral ischemia and then to the early reperfusion period. The temporal changes in gene profiles, cell subtypes, and microglial function were comprehensively analyzed using single-cell RNA sequencing. We identified 37,614 microglial cells and divided them into eight distinct subpopulations. Mic_home, Mic_pre1, and Mic_pre2 subpopulations were three clusters mainly composed of cells from the control samples, in which Mic_home was a homeostatic subpopulation characterized by high expression of Hpgd and Tagap, and Mic_pre1 and Mic_pre2 were two clusters with preliminary inflammatory activation characteristics marked by P2ry13 and Wsb1 respectively. Mic_M1L1 and Mic_M1L2 subpopulations exhibited M1-like polarization manifested by the upregulation of inflammatory genes after ischemic stroke, while the intrinsic heterogeneity on the level of inflammatory responses and neurotrophic support properties was observed. Moreover, we identified three unique clusters of cells with low inflammation levels. Mic_np1, Mic_np2, and Mic_np3 were characterized by high expression of Arhgap45, Rgs10, and Pkm respectively. However, these cells did not show significant M2-like characteristics and their classic microglia function was also attenuated. These subpopulations exhibited higher activation of neuropeptide functional pathways. At last, we performed cell-cell communication analysis and identified major couplings contributing to the interaction between microglia and other cell populations. In summary, our study elucidated the temporal heterogeneity of microglia in the acute phase of ischemic stroke, which may facilitate the identification of effective neuroprotective targets to curb ischemic damage at an early stage.

Dose-Response Relationship between Red Blood Cell Distribution Width and In-Hospital Mortality in Oldest Old Patients with Acute Ischemic Stroke

Introduction: It is crucial to identify predictors of mortality in the early stage of acute ischemic stroke for the oldest old (aged ≥80 years) because of their poor overall survival outcomes. However, limited data are available as the oldest old have often been excluded from previous clinical studies. Hence, we aimed to assess the predictive effect of red blood cell distribution width on in-hospital mortality and the dose-response relationship between the red blood cell distribution width and in-hospital mortality in oldest old with acute ischemic stroke. Methods: A retrospective cohort study was performed in two tertiary hospitals. Patients aged ≥80 years admitted due to acute ischemic stroke from January 1, 2014, to January 31, 2020, were included in the study. We divided the eligible patients into 3 groups with tertiles of red blood cell distribution width. Restrictive cubic spline and robust locally weighted regression analysis were performed to test the dose-response relationship between red blood cell distribution width and the in-hospital mortality risk. All-cause in-hospital mortality was the main study outcome. Results: Overall, 606 patients were included in the final analysis. Red blood cell distribution width was categorized into 3 groups (T1: <13.7%, T2: 13.8–15.7%, and T3: >15.7%). The rationality of this categorization was then validated with restricted cubic spline and robust locally regression smoothing scatterplot, respectively. After adjusting for demographic and clinical features, a higher red blood cell distribution width was independently associated with in-hospital mortality and the hazard ratio (HR) was 3.31 (95% CI 2.47–4.45, p < 0.001). There was a positive dose-response relationship between red blood cell distribution width and mortality risk. Sensitivity analysis identified no conspicuous change in the HR. Conclusions: Red blood cell distribution width may be a valuable and simple measure for predicting in-hospital mortality in oldest old patients with acute ischemic stroke.

STRUCTURAL CHANGES OF ORGANS AND TISSUES UNDER CONDITIONS OF DEHYDRATION DISORDERS OF WATER-SALT EXCHANGE

Most processes in the body take place with the participation of water. It is an indispensable component of living organisms. Any violations of the water- electrolyte balance have consequences for the structural organization of all organs and tissues. Studying changes in the structural organization of organs and tissues during dehydration is an urgent task of both theoretical and practical medicine. The aim was to analyze and generalize data from modern scientifi c literature on structural changes in organs and tissues during dehydration. Water is involved in maintaining a normal body temperature, in the absorption of nutrients in the gastrointestinal tract, in the excretion of harmful substances and toxins with urine and sweat, and in many other processes. According to the classifi cation, intracellular (hyperosmolar dehydration), extracellular (isosomolar), extracellular (hypoosmolar dehydration), and general dehydration are distinguished. There is evidence in modern scientifi c literature that dehydration leads to structural changes in the thyroid gland, stomach, tongue, kidneys, adrenal gland, salivary glands, larynx, brain and many others. Dehydration disrupts the process of bone tissue regeneration. Dehydration is associated with progression of infarct volume and decreased cerebral blood fl ow during the acute stage of ischemic stroke. Recurrent dehydration associated with chronic intermittent fluid intake accelerates the progression of chronic kidney disease and hypertension. Therefore, under the conditions of dehydration of the body, all the studied organs and tissues undergo changes, their severity depends on the degree of dehydration, species, environmental conditions and regime. At this time, there are no data on changes in the lymphoid organs, in particular the thymus and the spleen, under conditions of dehydration.

Network Pharmacology Analysis and Experimental Validation of Kaempferol in the Treatment of Ischemic Stroke by Inhibiting Apoptosis and Regulating Neuroinflammation Involving Neutrophils.

Kaempferol, a natural plant flavonoid compound, has a neuroprotective effect on ischemic stroke, while the specific mechanism remains unclear. In the current study, we applied the comprehensive strategy that combines network pharmacology and experimental evaluation to explore the potential mechanism of kaempferol in the treatment of cerebral ischemia. First, network pharmacology analysis identified the biological process of kaempferol, suggesting that kaempferol may partly help in treating ischemic stroke by regulating apoptosis and inflammatory response. Then, we evaluated the efficacy of kaempferol in the acute stage of ischemic stroke and elucidated its effects and possible mechanisms on cell apoptosis and neuroinflammation involved by neutrophils. The results showed that kaempferol could significantly reduce the modified neurological severity score (mNSS), and reduce the volume of cerebral infarction and the degree of cerebral edema. In terms of anti-apoptosis, kaempferol could significantly reduce the number of TUNEL-positive cells, inhibit the expression of pro-apoptotic proteins and promote the expression of anti-apoptotic proteins. Kaempferol may play an anti-apoptotic role by up-regulating the expression level of the BDNF-TrkB-PI3K/AKT signaling pathway. In addition, we found that kaempferol inhibited neuron loss and the activation of glial cells, as well as the expression level of the inflammatory protein COX-2 and the classic pro-inflammatory signaling pathway TLR4/MyD88/NF-κB in the ischemic brain, reduced MPO activity and neutrophil counts in peripheral blood, and down-regulated neutrophil aggregation and infiltration in the ischemic brain. Western blot revealed that kaempferol down-regulated the activation of the JAK1/STAT3 signaling pathway in neutrophils and ischemic brains. Our study showed that kaempferol inhibited the activation and number of neutrophils in the rat peripheral blood and brain, which may be related to the down-regulation of the JAK1/STAT3 pathway.

Elevation of neutrophil carcinoembryonic antigen-related cell adhesion molecule 1 associated with multiple inflammatory mediators was related to different clinical stages in ischemic stroke patients.

BackgroundWe aimed to analyze the level of carcinoembryonic antigen‐related cell adhesion molecule 1 (CEACAM1) in neutrophils of ischemic stroke (IS) patients at different stages, together with its roles in neutrophils.Patients and methodsSixty‐seven patients were classified into acute phase group (n = 19), subacute phase group (n = 28), and stable phase group (n = 20), and 20 healthy individuals who had received physical examination at the same time period as healthy control. We then analyzed the expression level of CEACAM1 and cell viability in CEACAM1 positive and CEACAM1 negative neutrophils by flow cytometry and the content of plasma CEACAM1, neutrophil gelatinase‐associated lipocalin (NGAL), matrix metalloproteinases‐9 (MMP‐9) was measured using enzyme‐linked immunosorbent assay (ELISA), while that of interleukin‐10 (IL‐10) and tumor necrosis factor (TNF) was determined using a Human Enhanced Sensitivity Flex set.ResultsCompared with healthy control, the percentage of CEACAM1 positive neutrophils in IS patients showed a significant increase, and a significant increase was also noticed in the content of plasma CEACAM1 at the subacute stage. Reduction in cell viability was observed in CEACAM1 positive neutrophils compared with CEACAM1 negative counterparts. There was a positive correlation between CEACAM1 expression rate in neutrophils and plasma CEACAM1 and IL‐10 content in the subacute group. Compared with acute group and healthy control group, there was an instinct increase in the level of plasma MMP‐9 and NGAL in subacute group.ConclusionsOur data showed that there was a rapid increase of CEACAM1 in neutrophils at the acute stage of IS. We speculated that CEACAM1 may serve as an inhibitory regulator involving in the progression of IS.

Serpine1 Regulates Peripheral Neutrophil Recruitment and Acts as Potential Target in Ischemic Stroke.

IntroductionPeripheral neutrophil infiltration can exacerbate ischemia–reperfusion injury. We focused on the relationship between various peripheral immune cells and cerebral ischemia–reperfusion (I/R) injury.MethodsIn this study, we investigated the effects of dauricine on neuronal injury induced by ischemia–reperfusion and peripheral immune cells after ischemic stroke in mouse model, and we explored the undefined mechanisms of regulating peripheral immune cells through RNA sequencing and various biochemical verification in vitro and in vivo.ResultsWe found that dauricine improved the neurological deficits of I/R injury, reduced the infarct volume, and improved the neurological scores. Furthermore, dauricine reduced the infiltration of neutrophils into the brain after MCAO-R and increased peripheral neutrophils but unchanged the permeability of the endotheliocyte Transwell system in an in vitro blood-brain barrier (BBB) model. RNA sequencing showed that chemotaxis factors, such as CXCL3, CXCL11, CCL20, CCL22, IL12a, IL23a, and serpine1, might play a crucial role. Overexpression of serpine1 reversed LPS-induced migration of neutrophils. Dauricine can directly bind with serpine1 in ligand–receptor docking performed with the Autodock and analyzed with PyMOL.ConclusionWe identified chemotaxis factor serpine1 played a crucial role in peripheral neutrophil infiltration, which may contribute to reduce the neuronal injury induced by ischemia–reperfusion. These findings reveal that serpine1 may act as a potential treatment target in the acute stage of ischemic stroke.