Excitotoxic Storms of Ischemic Stroke: A Non-neuronal Perspective.

Numerous neurological disorders share a fatal pathologic process known as glutamate excitotoxicity. Among which, ischemic stroke is the major cause of mortality and disability worldwide. For a long time, the main idea of developing anti-excitotoxic neuroprotective agents was to block glutamate receptors. Despite this, there has been little successful clinical translation to date. After decades of "neuron-centered" views, a growing number of studies have recently revealed the importance of non-neuronal cells. Glial cells, cerebral microvascular endothelial cells, blood cells, and so forth are extensively engaged in glutamate synthesis, release, reuptake, and metabolism. They also express functional glutamate receptors and can listen and respond for fast synaptic transmission. This broadens the thoughts of developing excitotoxicity antagonists. In this review, the critical contribution of non-neuronal cells in glutamate excitotoxicity during ischemic stroke will be emphasized in detail, and the latest research progress as well as corresponding therapeutic strategies will be updated at length, aiming to reconceptualize glutamate excitotoxicity in a non-neuronal perspective.

Progress on traditional Chinese medicine in treatment of ischemic stroke via the gut-brain axis.

Ischemic stroke is a common issue that severely affects the human health. Between the central nervous system and the enteric system, the " Gut-Brain " axis, the bidirectional connection involved in the neuro-immuno-endocrine network, is crucial for the occurrence and development of ischemic stroke. Ischemic stroke can lead to change in the gut microbiota and gastrointestinal hormones, which will then reversely affect the disease development. Traditional Chinese Medicine (TCM) has unique advantages with reference to the treatment for ischemic stroke. The latest research revealed that a significant portion of medicines and prescriptions of TCM exert their therapeutic effects by improving the gut microbiota and regulating the secretion of gastrointestinal hormones. The present review summarized the Chinese medicines that play a therapeutic role in cerebral ischemia through regulating the "Gut-Brain" axis and described the corresponding mechanisms. This study attempts to provide reference for clinical selection of Chinese medicines and helps better understand the relevant mechanisms of action.

A novel glycosylation-related gene signature predicts survival in patients with lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common malignant tumor that seriously affects human health. Previous studies have indicated that abnormal levels of glycosylation promote progression and poor prognosis of lung cancer. Thus, the present study aimed to explore the prognostic signature related to glycosyltransferases (GTs) for LUAD. METHODS: The gene expression profiles were obtained from The Cancer Genome Atlas (TCGA) database, and GTs were obtained from the GlycomeDB database. Differentially expressed GTs-related genes (DGTs) were identified using edge package and Venn diagram. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and ingenuity pathway analysis (IPA) methods were used to investigate the biological processes of DGTs. Subsequently, Cox and Least Absolute Shrinkage and Selection Operator (LASSO) regression analyses were performed to construct a prognostic model for LUAD. Kaplan-Meier (K-M) analysis was adopted to explore the overall survival (OS) of LUAD patients. The accuracy and specificity of the prognostic model were evaluated by receiver operating characteristic analysis (ROC). In addition, single-sample gene set enrichment analysis (ssGSEA) algorithm was used to analyze the infiltrating immune cells in the tumor environment. RESULTS: A total of 48 DGTs were mainly enriched in the processes of glycosylation, glycoprotein biosynthetic process, glycosphingolipid biosynthesis-lacto and neolacto series, and cell-mediated immune response. Furthermore, B3GNT3, MFNG, GYLTL1B, ALG3, and GALNT13 were screened as prognostic genes to construct a risk model for LUAD, and the LUAD patients were divided into high- and low-risk groups. K-M curve suggested that patients with a high-risk score had shorter OS than those with a low-risk score. The ROC analysis demonstrated that the risk model efficiently diagnoses LUAD. Additionally, the proportion of infiltrating aDCs (p < 0.05) and Tgds (p < 0.01) was higher in the high-risk group than in the low-risk group. Spearman's correlation analysis manifested that the prognostic genes (MFNG and ALG3) were significantly correlated with infiltrating immune cells. CONCLUSION: In summary, this study established a novel GTs-related risk model for the prognosis of LUAD patients, providing new therapeutic targets for LUAD. However, the biological role of glycosylation-related genes in LUAD needs to be explored further.

Research progress on astrocyte autophagy in ischemic stroke.

Ischemic stroke is a highly disabling and potentially fatal disease. After ischemic stroke, autophagy plays a key regulatory role as an intracellular catabolic pathway for misfolded proteins and damaged organelles. Mounting evidence indicates that astrocytes are strongly linked to the occurrence and development of cerebral ischemia. In recent years, great progress has been made in the investigation of astrocyte autophagy during ischemic stroke. This article summarizes the roles and potential mechanisms of astrocyte autophagy in ischemic stroke, briefly expounds on the crosstalk of astrocyte autophagy with pathological mechanisms and its potential protective effect on neurons, and reviews astrocytic autophagy-targeted therapeutic methods for cerebral ischemia. The broader aim of the report is to provide new perspectives and strategies for the treatment of cerebral ischemia and a reference for future research on cerebral ischemia.

[Acupuncture up-regulates MCT2 expression of peri-ischemic cortex in middle cerebral artery occlusion rats].

OBJECTIVE: To observe the effect of manual acupuncture (MA)＋ electroacupuncture (EA) on changes of neurological function and expression of monocarboxylate transporter 2(MCT2)in cerebral ischemia (CI) rats, so as to explore its mechanism underlying improvement of ischemic cerebrovascular disease. METHODS: Eighty male Wistar rats were equally randomized into four groups: normal control (normal), sham operation (sham), model and acupuncture. The CI model was induced by middle cerebral artery occlusion (MCAO) with a thread embolus. Manual acupuncture stimulation (mild twisting reinforcing-reducing method) was applied to "Baihui"(GV20)and "Fengfu"(GV16) for 10 min. EA (1 mA, 2 Hz /15 Hz) was respectively applied to bilateral "Quchi" (LI11) and "Zusanli"(ST36) for 20 min, once per day for 7 days. The neurological deficit severity was evaluated according to Zea Longa's methods. The activity of lactate dehydrogenase (LDH), fructose-6-phosphate kinase (PFK) and pyruvate kinase (PK) in the peri-ischemic cortex tissue was detected by enzymatic chemistry, and the expression of MCT2 detected by immunofluorescence histochemistry, Western blot and quantitative real-time PCR, separately. RESULTS: After CI and in comparison with the normal and sham groups, the Zea Longa's score, the fluorescence intensity and the expression level of MCT2 protein were significantly increased (P<0.01), and the activity of LDH, PFK and PK in the peri-ischemic cortex was significantly decreased in the model group (P<0.01). There was no significant change in the relative expression of MCT2 mRNA (P>0.05). Following the intervention and in comparison with the model group, the Zea Longa's score was considerably decreased in the acupuncture group (P<0.01), the activity of LDH, PFK and PK，and the expression levels of MCT2 protein and mRNA were considerably or further up-regulated in the acupuncture group (P<0.01, P<0.05). CONCLUSION: Acupuncture intervention can improve neurological function in CI rats, which is possibly related with its effects in up-regulating the expression of MCT2 and promoting the utilization of lactate in peri-ischemic cortex.

A pillar[5]arene-based multiple-stimuli responsive metal-organic gel was constructed for facile removal of mercury ions.

A thioacetohydrazide functionalized pillar[5]arene was synthesized, which could further assemble into a linear supramolecular metal-organic polymer upon adding Zn2+. Furthermore, the obtained linear supramolecular metal-organic polymer could self-assemble to form a fluorescent supramolecular metal-organic gel at high concentration. When TBAOH was added to the viscous solution at high temperature, the obtained solution could not form a supramolecular metal-organic gel upon cooling. More importantly, when Hg2+ ions are added to the metal-organic gel, the strong blue fluorescence is clearly quenched, and this metal-organic gel (xerogel) could effectively remove Hg2+ from water. Simultaneously, a thin film based on the metal-organic gel was prepared, which was confirmed to be a convenient test kit for detecting Hg2+.

Effects of Radix Astragali and Its Split Components on Gene Expression Profiles Related to Water Metabolism in Rats with the Dampness Stagnancy due to Spleen Deficiency Syndrome.

Radix Astragali (RA) with slight sweet and warm property is a significant "qi tonifying" herb; it is indicated for the syndrome of dampness stagnancy due to spleen deficiency (DSSD). The purpose of this research was to explore effects of RA and its split components on gene expression profiles related to water metabolism in rats with the DSSD syndrome for identifying components representing property and flavor of RA. The results indicated that RA and its split components, especially polysaccharides component, significantly increased the body weight and the urine volume and decreased the water load index of model rats. Our data also indicated differentially expressed genes (DEGs) related to water metabolism involved secretion, ion transport, water homeostasis, regulation of body fluid levels, and water channel activity; the expression of AQP1, AQP3, AQP4, AQP5, AQP6, and AQP8 was improved; calcium, cAMP, MAPK, PPAR, AMPK, and PI3K-Akt signaling pathway may be related to water metabolism. In general, results indicate that RA and its split components could promote water metabolism in rats with the DSSD syndrome via regulating the expression of AQPs, which reflected sweet-warm properties of RA. Effects of the polysaccharides component are better than others.

Accessory spleen appearing as an intrasplenic pseudo-tumoral mass: A rare case report.

The current study presents a rare case of an accessory spleen that manifested as a solid intrasplenic pseudotumor. The affected patient was previously healthy. Upon examination with computed tomography (CT), an ovoid, soft-tissue mass of ~4.1 cm in diameter was found on the upper pole of the spleen. Biochemical indices, such as blood routine and coagulation tests, and tumor marker analysis, revealed no abnormalities. Another CT scan was performed, but this failed to indicate whether the mass was benign or malignant. Therefore, the lesion was resected along with the spleen by laparoscopic surgery. The resected sample was subject to pathological examinations for final validation, and was finally diagnosed as an accessory spleen. The patient was followed up for six months with no signs of recurrence.

[Effect of electroacupuncture intervention on neurological function, cerebral blood flow and cerebral cytochrome P 450 2 C 11 mRNA expression in rats with focal cerebral ischemia].

OBJECTIVE: To observe the effect of electroacupuncture(EA) intervention on changes of neurological function and expression of cerebral cytochrome P 450 2 C 11 (CYP 2 0 11) mRNA in focal cerebral ischemia (FCI) rats, so as to explore its mechanism underlying improvement of ischemic cerebral vascular disease. METHODS: Wistar rats were randomly divided into normal, model, EA and EA+ 17-ODYA (17-Octadecynoic acid, an inhibitor for the metabolism of arachidonic acid by cytochrome P 450) groups. Focal cerebral ischemia rats were induced by middle cerebral artery occlusion (MCAO) with thread embolus. EA was applied to bilateral "Neiguan"(PC 6 ) and "Quchi" (LI 11) after MCAO. Zea Longa's score and beam walking test (BWT) score of rats were used to evaluate the neurological impairment. Local cerebral blood flow (LCBF) of the pial tissue was moni- tored using Laser-Doppler Flowmetry. The expression of cerebral CYP 2 C 11 mRNA was examined by Real-time Quantitative PCR (qPCR). RESULTS: In comparison with the normal group, BWT score and LCBF of the model group were significantly decreased (P<0.05), and Zea Longa's score and cerebral CYP 2 C 11 mRNA expression level of the model group were significantly increased (P<0.05). While in comparison with the model group, BWT score, LCBF and cerebral CYP2 C 11 mRNA levels were considerably up-regulated and Zea Longa's score was down-regulated in the EA group (P<0.05) rather than in the EA + 17- ODYA group (P<0.05). H.E. stainshowed that the nerve impairment of the ischemic cerebral tissue including the neuronal degeneration, necrosis, apoptosis, etc. was relatively milder in the EA group. CONCLUSION: EA intervention can improve cerebral blood flow and up-regulate cerebral CYP 2 C 11 mRNA expression in FCI rats, which may contribute to its action in improving neu- rological impairment.

[Effects of electroacupuncture intervention on neurological function, blood glucose and insulin levels in rats with focal cerebral ischemia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on dynamic changes of neurological function, blood glucose and insulin levels in rats with focal cerebral ischemia (CI), so as to explore its mechanism underlying improvement of ischemic cerebral vascular disease. METHODS: Male Wistar rats were randomly divided into four groups: normal, sham-operation (sham), model and EA. CI model was induced by middle cerebral artery occlusion (MCAO). The four groups were further randomized into 5 subgroups according to time-points of MCAO: 6 hour, day 1, day 3, day 7 and day 14 (8 rats/ group). EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Neiguan" (PC 6) and "Quchi" (LI 11) for 30 min, once daily for 1 day to 14 days. Neurological impairment was evaluated by Zea Longa 5-point scoring system. Local cerebral blood flow (LCBF) of the cerebral pia mater was determined using Laser Doppler Flowmetry. The levels of abdominal cavity venous blood glucose and serum insulin were measured by using a glucose meter and radioimmunoassay, respectively. RESULTS: In comparison with the normal and sham groups, the rats' body weights and LCBF levels at time-points of day 1, 3, 7 and 14, and blood glucose and serum insulin levels at most time-points of 6 h, day 1, 3, 7 and 14 after CI were significantly decreased in the model group (P < 0.05), and neurological scores at time-points of 6 h, day 1, 3, 7 and 14 were markedly increased in the model group (P < 0.05). After EA intervention, compared with the model group, the rats' body weights at time points of day 3, 7 and 14, LCBF levels on day 7 and 14, blood glucose on day 14, and serum insulin contents at time-points of day 1, 3, 7 and 14 were considerably increased in the EA group (P < 0.05), while Zea Longa scores at time-points of day 3 and 7 were evidently decreased in the EA group (P<0.05). CONCLUSION: EA intervention is effective in improving neurological function in CI rats, which is probably associated with its functions in improving cerebral blood flow, and up-regulating blood glucose and insulin levels.

[Construction of a recombinant shRNA expression vector targeting EZH2 gene and its inhibitory effect on colon adenocarcinoma SW480 cells].

OBJECTIVE: To construct a recombinant short hairpin RNA (shRNA) expression vector targeting EZH2 gene, and to determine its effect on the proliferation of colon adenocarcinoma SW480 cells. METHODS: The DNA sequence with short hairpin structure was designed according to the EZH2 cDNA sequence and cloned into PGFP-V-RS vector to construct a recombinant expression vector silencing EZH2 gene. After identification, the shRNA-expressing vector was then transfected into SW480 cells. RT-PCR and Western blot were used to detect the inhibitory effect at both mRNA and protein levels. MTT was used to detect cell viability due to the alteration of EZH2 gene activity. RESULTS: At 48 h after transfection, the expression of EZH2 mRNA in the gene silencing group and negative control group were 0.339 ± 0.013 and 1.968 ± 0.072, respectively. The expression of EZH2 protein in the gene silencing group and negative control group were 0.229 ± 0.008 and 1.168 ± 0.053, respectively. The expression of EZH2 in the gene silencing group was significantly lower than that in the negative control group (P < 0.01, P < 0.05). At 48 and 72 h after transfection, the inhibition rate of cell growth in the gene silencing group was 30.7% and 25.9%, respectively, indicating that the cell growth was significantly inhibited in comparison with that in the blank control group (P < 0.05). CONCLUSIONS: A recombinant shRNA expression vector targeting EZH2 gene has been successfully constructed in this study, with a significant inhibitory effect on the proliferation of SW480 cells. This lays an experimental foundation for further exploring the mechanism underlying the action of EZH2 gene on tumor biology.

[Effects of acetoacetate extract of Radix Aconite on hepatic contents of LA, LDH, PA, Gn and ATPase activities in deficient cold model rats].

OBJECTIVE: To observe different effects of acetoacetate extract of Radix Aconite and Radix Aconite Decoction on the energy metabolism in deficient cold model rats. METHODS: Wistar rats were randomly divided into the blank control group (n=10) and the deficient cold model group (n=30). The deficient cold rat model was established using decoction consisting of gypsum, Radix Gentianae, Cortex Phellodendri, and Rhizoma Anemarrhenae. The decoction was given to rats of the deficient cold model group by gastrogavage for 5 days. Then these rats were randomly divided into the acetoacetate extract of Radix Aconite group (n=10), the Radix Aconite Decoction group (n=10), and the model group (n=10). Rats in the model model group were administered with the decoction by gastrogavage. Rats in the other two groups were administered with the acetoacetate extract of Radix Aconite or Radix Aconite Decoction by gastrogavage for 5 days. The contents of lactic acid (LA), lactate dehydrogenase (LDH), pyruvate (PA), glycogen (Gn) and activities of Na(+) -K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase in the hepatic tissue were detected. RESULTS: Compared with the blank control group, the PA content, activities of Na(+)-K(+) -ATPase and Ca(2+) -Mg(2+) -ATPase decreased in the model group. Compared with the model group, the PA content increased in the other two groups. Compared with the control group, the contents of LDH and PA, and activities of Na(+) -K(+) -ATPase increased in the the acetoacetate extract of Radix Aconite group with statistical difference (P < 0.05). CONCLUSIONS: The febricity of acetoacetate extract of Radix Aconite was slightly higher than that of Radix Aconite Decoction, seemingly generating more energy. But the final conclusions and concrete mechanisms of action need further studies.