Wei Zou

Drug-resistant epilepsy (DRE), also known as intractable epilepsy or refractory epilepsy, is a disease state with long-term poorly controlled seizures attack. Without effective treatment, patients are at an elevated risk of injury, premature death, mental disorders, and poor quality of life, increasing the need for a fresh perspective on the etiology and treatment of DRE. The gut is known to harbor a wide variety of microorganisms that can regulate the host’s response to exogenous signals and participate in various physiological and pathological processes in the human body. Interestingly, emerging evidence has uncovered the changes in gut microbiota in patients with epilepsy, particularly those with DRE. In addition, both dietary interventions and specific antibiotic therapy have been proven to be effective in restoring the microecological environment and, more importantly, reducing seizures. Here, we reviewed recent studies on DRE and the involvement of gut microbiota in it, describing changes in the gut microflora composition in patients with DRE and corresponding animal models. Furthermore, the influence of the ketogenic diet, probiotics, fecal microbiota transplantation (FMT), and antibiotics as microbiome-related factors on seizure control and its possible mechanisms are broadly discussed. Finally, we highlighted the significance of gut microbiome in DRE, in order to provide a new prospect for early identification and individualized treatment of patients with DRE.

We used an in vitro model of the human brain immune microenvironment to simulate hypoxic-ischemic brain injury (HIBI) and treatment with human umbilical cord mesenchymal stem cells (hUMSCs) to address the transformation barriers of gene differences between animals and humans in preclinical research. A co-culture system, termed hNAME, consisted of human hippocampal neurons (N), astrocytes (A), microglia (M), and brain microvascular endothelial cells (E). Flow cytometry measured the apoptosis rates of neurons and endothelial cells. hNAME-neurons and endothelial cells experienced more severe damage than monolayer cells, particularly after 48 h and 24 h of reoxygenation (OGD48/R24). Western blotting identified neuroinflammatory response markers, including HIF-1α, C1q, C3, TNF-α, and iNOS. Inflammatory factors originated from the glial chamber rather than the neurons and vascular endothelial chambers. A gradual increase in the release of inflammatory factors was observed as the OGD and reoxygenation times increased, peaking at OGD48/R24. The hNAME value was confirmed in human umbilical cord mesenchymal stem cells (hUMSCs). Treatment with hUMSCs resulted in a notable decrease in the severity of neuronal and endothelial cell damage in hNAME. The hNAME is an ideal in vitro model for simulating the immune microenvironment of the human brain because of the interactions between neurons, vessels, astrocytes, and microglia.

Glioblastoma (GBM) is a deadly and common primary brain tumor. Poor prognosis is linked to high proliferation and cell heterogeneity. Sex differences may play a role in patient outcome. Previous studies showed that ER‐α36, a variant of the estrogen receptor (ER), mediated non‐genomic estrogen signaling and is highly expressed in many ER‐negative malignant tumors. ER‐α36 also associates with epidermal growth factor receptor (EGFR). The primary purpose of this study is to investigate the cross talk between ER‐α36 and EGFR in estrogen‐mediated GBM cell proliferation. Here, we showed that ER‐α36 was highly expressed and confirmed that ER‐α36 co‐labels with EGFR in human GBM samples using immunohistochemical techniques. We also investigated the mechanisms of estrogen‐induced proliferation in ER‐α‐negative cell lines. We found that GBM cells showed varying responsive to mitogenic estrogen signaling which correlated with ER‐α36 expression, and knockdown of ER‐α36 diminished the response. Exposure to estrogen also caused upregulation of cyclin protein expression in vitro. We also found that low concentration of estrogen promoted SRC‐Y‐416 and inhibited SRC‐Y‐527 phosphorylation, corresponding with activated SRC signaling. Inhibiting SRC or EGFR abolished estrogen‐induced mitogenic signaling, including cyclin expression and MAPK phosphorylation. Cumulatively, our results demonstrate that ER‐α36 promotes non‐genomic estrogen signaling via the EGFR/SRC/MAPK pathway in GBM. This may be important for the treatment of ER‐α‐negative GBMs that retain high level of ER‐α36, since estrogen may be a viable therapeutic target for these patients.

Objectives: To explore the possible mechanism of human umbilical cord mesenchymal stem cell (hUC-MSC) transplantation in mice after spinal cord hemisection. Methods: Thoracic spinal cord hemisection injuries were performed on adult female Kunming mice. The mice with spinal cord injury (SCI) were injected with hUC-MSCs suspended in normal saline, while the control mice received an equal volume of normal saline. The histological HE staining and Nissl staining were performed 4 and 8 weeks after hUC-MSC transplantation in SCI mice. The Basso-Beattie-Bresnahan (BBB) locomotor rating scale was used to assess functional recovery after SCI. Western blotting was performed to determine the protein expressions. Results: hUC-MSCs transplantation decreased cavitation and tissue loss and increased the number of Nissl bodies in the damaged areas of the spinal cord after 4 and 8 weeks. The BBB locomotor performance of the transplanted mice was significantly improved (P<0.01). The wet weight of the injured side of the gastrocnemius muscle was significantly higher in the transplant group than that in the control group. Western blotting showed that TUJ1 and Olig2 expressions were significantly higher in hUC-MSC-grafted mice than those in vehicle controls. Three days after hUC-MSC transplantation, the expressions of TNF-alpha and NF-kB were higher in MSC-grafted mice than those in vehicle controls. However, 4 weeks after stem cell transplantation, the expressions of these two factors decreased in hUC-MSC-grafted mice compared with those in the vehicle controls. At 8 weeks after hUC-MSC transplantation, the expression of PTBP-1 was decreased in hUC-MSC-grafted mice compared with that in vehicle controls. Conclusions: hUC-MSC transplantation can protect neuron survival, promote myelin repair, and control glial scar formation in SCI mice.

Glioblastoma is a deadly and common primary brain tumor. Poor prognosis is linked to high proliferation and cell heterogeneity. Sex differences may play a role in patient outcome. Previous studies showed that ER-α36, a variant of the estrogen receptor, mediated non-genomic estrogen signaling and is highly expressed in many ER-negative tumors. ER-α36 also associates with the EGFR. We showed that ER-α36 was highly expressed and confirmed that ER-α36 co-labels with EGFR in glioma specimens. We also investigated the mechanisms of estrogen induced proliferation in ER-α-negative cells U87 and U251. We found that glioblastoma cell varying responsive to mitogenic estrogen signaling which correlated with ER-α36 expression, and knockdown of ER-α36 diminished the response. Exposure to estrogen also caused up-regulation of cyclin protein expression in vitro. We also found that low concentrations of estrogen promoted SRC-Y-416 and inhibited SRC-Y-527 phosphorylation, corresponding with activated SRC signaling. Inhibiting SRC or EGFR abolished estrogen-induced mitogenic signaling, including cyclin expression and MAPK phosphorylation. Cumulatively, our results demonstrate that ER-α36 promotes non-genomic estrogen signaling via the EGFR/SRC/MAPK pathway in glioblastoma. This may be important for the treatment of ER-α-negative glioblastomas that retain high level of ER-α36, since estrogen may be a viable therapeutic target for these patients.

Flor·Essence (FE), a natural food grade herbal formula product manufactured by Flora Manufacturing & Distributing Ltd., has been used by patients with cancer in North America to stimulate immune cells in order to attenuate or reverse immune damage. To elucidate the mechanisms underlying the effects of FE on the immune system, spleen lymphocyte proliferation was analyzed by an MTT assay, and the phagocytic capacity of macrophages was measured via the neutral red phagocytosis method. The cytotoxicity of natural killer (NK) cells towards K562 cells was assessed via a CytoTox 96 assay. The production of the cytokines interleukin (IL)‑12 and interferon (IFN)‑γ in the peripheral blood was determined via ELISA and PCR analysis. The expression levels of caveolin‑1 and NF‑κB were measured via western blotting. In addition, cyclophosphamide was used to establish a mouse model of immunosuppression. It was found that the proliferation of splenocytes, the phagocytic capacity of macrophages and the cytotoxicity of NK cells against K562 cells were increased after oral administration of FE to mice. FE augmented the production of IL‑12 and IFN‑γ in the peripheral blood of mice. FE significantly increased the expression of proliferating cell nuclear antigen and caveolin‑1, and decreased NF‑κB expression. Finally, FE enhanced the viability of immune cells from cyclophosphamide‑treated immunosuppressed mice. The results indicated that FE could activate immune responses and enhance natural immunity, suggesting that oral administration of FE can activate the body's immune response and resist damage caused by cyclophosphamide chemotherapy.

Aberrant oxidative metabolism in cells is one of the hallmarks of cancer. Overproduction of reactive species promotes carcinogenesis by inducing genetic mutations and activating oncogenic pathways, and thus, antioxidant therapy is considered as an important strategy for cancer prevention and treatment. Caveolin-1 (Cav-1), a constituent protein of caveolae, is involved in not only the formation of the caveolae, vesicular transport, maintaining cholesterol homeostasis directly, but also many cellular physiological and pathological processes including growth, regulation of mitochondrial antioxidant level, apoptosis and carcinomas by interacting with a lot of signaling molecules through caveolin scaffolding domain. Cav-1 has also been shown to mediate tumor genesis and progression through oxidative stress modulation, while Cav-1-targeted treatment could scavenge the reactive species. Intracellular reactive species could modulate the expression, degradation, post-translational modifications and membrane trafficking of Cav-1. More importantly, emerging evidence has indicated that multiple antioxidants could exert antitumor activities in cancer cells by modulating the signaling of Cav-1. This paper reviewed the research progresses on the roles of Cav-1 and oxidative stress in tumorigenesis and development, and would provide new insights on designing strategies for cancer prevention or treatment.

K⁺ channels regulate a multitude of biological processes and play important roles in a variety of diseases by controlling potassium flow across cell membranes. They are widely expressed in the central and peripheral nervous system. As a malignant tumor derived from nerve epithelium, glioma has the characteristics of high incidence, high recurrence rate, high mortality rate, and low cure rate. Since glioma cells show invasive growth, current surgical methods cannot completely remove tumors. Adjuvant chemotherapy is still needed after surgery. Because the blood-brain barrier and other factors lead to a lower effective concentration of chemotherapeutic drugs in the tumor, the recurrence rate of residual lesions is extremely high. Therefore, new therapeutic methods are needed. Numerous studies have shown that different K⁺ channel subtypes are differentially expressed in glioma cells and are involved in the regulation of the cell cycle of glioma cells to arrest them at different stages of the cell cycle. Increasing evidence suggests that K⁺ channels express in glioma cells and regulate glioma cell behaviors such as cell cycle, proliferation and apoptosis. This review article aims to summarize the current knowledge on the function of K⁺ channels in glioma, suggests K⁺ channels participating in the development of glioma.

Glioblastoma (GBM) is a highly infiltrative and malignant primary brain tumor. Despite aggressive therapy, patients with GBM have a dismal prognosis with median survival of approximately 1 year. Tamoxifen (TAM), a selective estrogen receptor modulator (SERM), has been used to treat GBM for many years. ER‐α36 is a novel variant of estrogen receptor‐alpha66 (ER‐α66) and can mediate cell proliferation through estrogen or anti‐estrogen signaling in different cancer cells. Previously, we found that ER‐α36 was highly expressed in GBM and was involved in the tamoxifen sensitivity of glioblastoma cells. However, the molecular mechanism responsible has not been well established. Here, we found that ER‐α36 is highly expressed in glioblastoma specimens. We further found that ER‐α36 knockdown increased sensitivity of glioblastoma U87 cells to TAM and decreased autophagy in these cells. However, ER‐α36 overexpression decreased TAM sensitivity and induced autophagy. We also established TAM‐resistant glioblastoma U251 cells by a long‐term culture in TAM‐containing medium and found that TAM‐resistant cells showed a six‐fold increase of ER‐α36 mRNA expression and elevated basal autophagy. ER‐α36 knockdown in these TAM‐resistant cells restored TAM sensitivity. In addition, we recapitulated the physiologically relevant tumor microenvironment in an integrated microfluidic device, and U87 cells were treated with a gradient of TAM. We found that ER‐α36 expression is consistent with autophagy protein P62 in a three‐dimensional microenvironment. In summary, these results indicate that ER‐α36 contributes to tamoxifen resistance in glioblastoma cells presumably through regulation of autophagy.