Luping Zhang

Neural Stem Cells Enhance Nerve Regeneration after Sciatic Nerve Injury in Rats

With the development of tissue engineering and the shortage of autologous nerve grafts in nerve reconstruction, cell transplantation in a conduit is an alternative strategy to improve nerve regeneration. The present study evaluated the effects and mechanism of brain-derived neural stem cells (NSCs) on sciatic nerve injury in rats. At the transection of the sciatic nerve, a 10-mm gap between the nerve stumps was bridged with a silicon conduit filled with 5 × 105 NSCs. In control experiments, the conduit was filled with nerve growth factor (NGF) or normal saline (NS). The functional and morphological properties of regenerated nerves were investigated, and expression of hepatocyte growth factor (HGF) and NGF was measured. One week later, there was no connection through the conduit. Four or eight weeks later, fibrous connections were evident between the proximal and distal segments. Motor function was revealed by measurement of the sciatic functional index (SFI) and sciatic nerve conduction velocity (NCV). Functional recovery in the NSC and NGF groups was significantly more advanced than that in the NS group. NSCs showed significant improvement in axon myelination of the regenerated nerves. Expression of NGF and HGF in the injured sciatic nerve was significantly lower in the NS group than in the NSCs and NGF groups. These results and other advantages of NSCs, such as ease of harvest and relative abundance, suggest that NSCs could be used clinically to enhance peripheral nerve repair.

SRPK1 Dissimilarly Impacts on the Growth, Metastasis, Chemosensitivity and Angiogenesis of Glioma in Normoxic and Hypoxic Conditions

Glioma is among the ten most common causes of cancer-related death and has no effective treatment for it, so we are trying to find a new target for anticancer treatment. This study investigates the different expression of SRPK1 as a novel protein in glioma, which can influence tumor cells biological characteristics in normoxic and hypoxic environment. The expression levels of SRPK1 protein in glioma cell lines transfected with siSRPK1 or not were examined using immunofluorescence, RT-PCR and Western blot analysis, respectively. The impact of SRPK1 on the biological characteristics of U251 cells was further studied using methylthiazol tetrazolium assays, flow cytometry, and Transwell invasion chamber assays. The results showed that knockdown of SRPK1 inhibited tumor cells growth, invasion and migration in normoxic condition, but portion of the effect could be reversed in hypoxia. SRPK1 expression was induced in glioma cells by DDP treated, but not TMZ, in both normoxia and hypoxia conditions. We propose SRPK1 as a new molecular player contributing to the early treatment of glioma.

The influence of SRPK1 on glioma apoptosis, metastasis, and angiogenesis through the PI3K/Akt signaling pathway under normoxia

Gliomas, the most common primary brain tumors, have low survival rates and poorly defined molecular mechanisms to target for treatment. Serine/arginine SR protein kinases 1 (SRPK1) can highly and specifically phosphorylate the SR protein found in many tumors, which can influence cell proliferation and angiogenesis. However, the roles and regulatory mechanisms of SRPK1 in gliomas are not understood. The aim of this study was to determine the functions and regulation of SRPK1 in gliomas. We found that SRPK1 inhibition induces early apoptosis and significantly inhibits xenograft tumor growth. Our results indicate that SRPK1 affects Akt and eIF4E phosphorylation, Bax and Bcl-2 activation, and HIF-1 and VEGF production in glioma cells. Moreover, transfection of SRPK1 siRNA strongly reduced cell invasion and migration by regulating the expression of MMP2 and MMP9 and significantly decreased the volume of tumors and angiogenesis. We show here that a strong link exists among SRPK1, Akt, eIF4E, HIF-1, and VEGF activity that is functionally involved in apoptosis, metastasis, and angiogenesis of gliomas under normoxic conditions. Thus, SRPK1 may be a potential anticancer target to inhibit glioma progression.

The preparation and comparison of decellularized nerve scaffold of tissue engineering

To integrate tissue engineering concepts into strategies to repair spinal cord injury (SCI) has been a hotspot in recent years, and the choice of scaffolding material is crucial to tissue engineering. Recently, decellularized nerve scaffold becomes a central concern due to its peculiar superiority. In this study, the decellularized nerve scaffold was prepared with three different methods and a comparison was made to acquire an ideal scaffold materials. All sciatic nerves from Sprague-Dawley (SD) rats were randomly divided into four groups: A: normal control group, B: TritonX-100 with sodium deoxycholate group, C: TritonX-100 with enzyme group and D: freezing-thawing with enzyme group. Histology and transmission electron microscope were exploited to evaluate the effect of removing cells and immunological histological chemistry was exploited to evaluate immunogenicity. Meanwhile the mechanical properties were evaluated by mechanics index. Hematoxylin and eosin (HE) staining and electron microscopic examinations reveal that the cell components and myelin sheaths are the least in the freezing-thawing with enzyme group. Immunohistochemistry shows that the immunogenicity is lower in group B, C, and D than the control group, and the group D has the lowest immunogenicity. Mechanical testing shows that there is no significant difference after acellular processing. Sciatic nerve, cell-extracted by freezing-thawing with enzyme, could obtain the ideal scaffold materials which has no cells and myelin sheaths. In addition, the decellularized nerve scaffold has no immunogenicity and the mechanical property of normal sciatic nerve is preserved.

mir-218-2 promotes glioblastomas growth, invasion and drug resistance by targeting CDC27

Glioma has become a significant global health problem with substantial morbidity and mortality, underscoring the importance of elucidating its underlying molecular mechanisms. Recent studies have identified mir-218 as an anti-oncogene; however, the specific functions of mir-218-1 and mir-218-2 remain unknown, especially the latter. The objective of this study was to further investigate the role of mir-218-2 in glioma. Our results indicated that mir-218-2 is highly overexpressed in glioma. Furthermore, we showed that mir-218-2 is positively correlated with the growth, invasion, migration, and drug susceptibility (to β-lapachone) of glioma cells. In vitro, the overexpression of mir-218-2 promoted glioma cell proliferation, invasion, and migration. In addition, the overexpression of mir-218-2 in vivo was found to increase glioma tumor growth. Accordingly, the inhibition of mir-218-2 resulted in the opposite effects. Cell division cycle 27 (CDC27), the downstream target of mir-218-2, is involved in the regulation of glioma cells. Our results indicate that the overexpression of CDC27 counteracted the function of mir-218-2 in glioma cells. These novel findings provide new insight in the application of mir-218-2 as a potential glioma treatment.

Plexin-B1 indirectly affects glioma invasiveness and angiogenesis by regulating the RhoA/αvβ3 signaling pathway and SRPK1.

Gliomas are one of the most common primary brain tumors in adults. They display aggressive invasiveness, are highly vascular, and have a poor prognosis. Plexin-B1 is involved in numerous cellular processes, especially cellular migration and angiogenesis. However, the role and regulatory mechanisms of Plexin-B1 in gliomas are not understood and were thus investigated in this study. By using multiple and diverse experimental techniques, we investigated cell apoptosis, mitochondrial membrane potential, cell migration and invasion, angiogenesis, PI3K and Akt phosphorylation, and also the levels of SRPK1 and αvβ3 in glioma cells and animal glioma tissues. The results indicated that Plexin-B1 expression in glioma cell lines is increased compared to normal human astrocytes. Plexin-B1 mediates RhoA/integrin αvβ3 involved in the PI3K/Akt pathway and SRPK1 to influence the growth of glioma cell, angiogenesis, and motility in vitro and in vivo. Thus, Plexin-B1 signaling regulates the Rho/αvβ3/PI3K/Akt pathway and SRPK1, which are involved in glioma invasiveness and angiogenesis. Therefore, the new drug research should focus on Plexin-B1 as a target for the treatment of glioma invasion and angiogenesis.

The expression patterns of Mis1 is related with the glioma grade and the cytoplasmic Mis1 promotes angiogenesis

To investigate the expression patterns of Msi1 and its relationship with tumour grading and angiogenesis in human glioma. Msi1 expression and the angiogenesis labelled by CD31 were detected by immunohistochemical staining in glioma from Grade I to Grade IV. MVD-CD31 was counted respectively in nuclear and cytoplasmic positive areas of Msi1. There was weak expression of Msi1 in the adjacent normal brain tissue of glioma, and there were less MVD-CD31 than in glioma. Glioma showed strong expression and different patterns of Msi1. From Grade I to Grade IV glioma, the expression pattern of Msi1 was changed from nuclear into cytoplasmic pattern gradually (P<0.05), and the expression patterns of Msi1 were related with the angiogenesis of glioma (P<0.05). It suggested that cytoplasmic Msi1 promoted angiogenesis and nuclear Msi-1 inhibited angiogenesis in glioma (P<0.05). Msi1 has different expression patterns in glioma from Grade I to Grade IV, and the different expression patterns play diverse roles in angiogenesis of glioma. This might explain that from Grade I to IV, glioma showed worse and worse prognosis.

The effects of Weichangshu tablet on intracellular Ca2+ concentration in cultured rat gastrointestinal smooth muscle cells

The objective of this study was to explore the effects of Weichangshu tablets on free Ca2+ concentration of the gastrointestinal smooth muscle cells of rats and the molecular mechanism of the Weichangshu tablets. Cultured SD rat gastrointestinal smooth muscle cells were divided into control group, Cisapride group, Weichangshu group, and control + ethylene glycol tetraacetic acid (EGTA) group, Cisapride + EGTA group, and Weichangshu + EGTA group. Laser scanning microscope and spectrophotometer detection were applied to detect the calcium concentration. The calcium concentrations in Weichangshu group and Cisapride group significantly increased vs. control, and those in Weichangshu group were higher than those in Cisapride group. Ca2+ concentration in Weichangshu + EGTA group, Cisapride + EGTA group vs. control + EGTA group were not significantly different. Weichangshu could increase gastrointestinal smooth muscle free Ca2+ concentration, and this role may be achieved through the promotion of cells within the flow of calcium ions.