Norcantharidin induces growth inhibition and apoptosis of glioma cells by blocking the Raf/MEK/ERK pathway.

BACKGROUND: Malignant gliomas represent the most common primary brain tumors. The prognosis of patients with malignant gliomas is poor in spite of current intensive therapy and novel therapeutic modalities are needed. Here we report that norcantharidin is effective in growth inhibition of glioma cell lines in vitro. METHODS: Glioma cell lines (U87 and C6) were treated with norcantharidin. The effects of norcantharidin on the proliferation and apoptosis of glioma cells were measured by 3-[4,5-dimethylthiazol-2-thiazolyl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and flow cytometry. Western blotting was employed to determine the signaling pathway changes. RESULTS: The results showed that norcantharidin effectively inhibited cell growth and induced apoptosis in glioma cells, which was concurrent with inhibition of the expression of phospho-MEK and phospho-ERK. Furthermore, the expression anti-apoptotic proteins Bcl-2 and Mcl-1 significantly reduced, but no changes in Bcl-xL and Bax. CONCLUSIONS: Our findings demonstrate that norcantharidin is effective for growth inhibition of glioma cell lines and suggest that norcantharidin may be a new therapeutic option for patients with glioma.

Endovascular treatment of chronic, recurrent headache secondary to chronic cerebral venous sinus thrombosis.

Headache is a clinical diagnosis linked to a number of medical and surgical disorders. A common etiology has not yet been established. It would seem that these cases can be related to some degree of cerebral venous outflow obstruction. We report 2 cases of chronic superior sagittal sinus thrombosis causing isolated intracranial hypertension. The patients were treated with intrasinus thrombolytic therapy.

Reconstructive endovascular treatment of vertical stenosis associated with adjacent aneurysm at the same arterial anatomic segment.

One case had a symptomatic vertebral artery stenosis coupled with a coincidental unruptured cerebral aneurysm at the same arterial anatomic segment. And another case had an asymptomatic vertebral artery stenosis coupled with a ruptured cerebral aneurysm at the same arterial anatomic segment. They underwent intracranial stenting. Both lesions were treated successfully and neither complications nor strokes occurred after the procedures. Covered stent placement in an intracranial stenosis with an adjacent ruptured or unruptured aneurysm may be a feasible method.

Emergent angioplasty and stent placement recanalization without thrombolysis in acute middle cerebral artery occlusions.

A significant proportion of patients with infarcts from large-vessel lesions have shown a poor response to systemic thrombolysis. Stents have been used to recanalize occluded or severely stenosed intracranial arteries in patients with acute stroke. This study evaluated the feasibility, efficacy, and safety of intracranial artery recanalization for acute middle cerebral artery (MCA) occlusion using emergent angioplasty and stent placement without thrombolysis. All patients from a retrospectively collected database who met the inclusion criteria and were treated with an intracranial stent for acute MCA occlusion were included. Treatment comprised angioplasty and stenting without interventional thrombolytic therapy. Recanalization was assessed by angiography immediately after stent placement based on the Thrombolysis in Myocardial Infarction (TIMI) score. Complications related to the procedure and outcomes were assessed. Neurologic status was evaluated before and after treatment. Eleven patients were treated with emergent angioplasty and stent placement. Partial or complete recanalization (TIMI 2 and 3) was achieved in 11 patients (100%) assessed by digital subtraction angiography immediately after MCA stenting. One patient died due to reocclusion of MCA 2 days after the procedure. Among the survivors, 7 patients (70%) had a good outcome (modified Rankin Scale score, 0-2) and 3 patients (30%) had a moderate outcome (modified Rankin Scale score, 3). Follow-up computed tomography angiography or magnetic resonance angiography revealed mild restenosis in 2 of the 10 patients. This preliminary experience demonstrates the technical feasibility and high rate of recanalization with emergent angioplasty and stenting without thrombolysis in patients with acute MCA occlusion.

Long non-coding RNA MEG3 regulates autophagy after cerebral ischemia/reperfusion injury.

Severe cerebral ischemia/reperfusion injury has been shown to induce high-level autophagy and neuronal death. Therefore, it is extremely important to search for a target that inhibits autophagy activation. Long non-coding RNA MEG3 participates in autophagy. However, it remains unclear whether it can be targeted to regulate cerebral ischemia/reperfusion injury. Our results revealed that in oxygen and glucose deprivation/reoxygenation-treated HT22 cells, MEG3 expression was obviously upregulated, and autophagy was increased, while knockdown of MEG3 expression greatly reduced autophagy. Furthermore, MEG3 bound miR-181c-5p and inhibited its expression, while miR-181c-5p bound to autophagy-related gene ATG7 and inhibited its expression. Further experiments revealed that mir-181c-5p overexpression reversed the effect of MEG3 on autophagy and ATG7 expression in HT22 cells subjected to oxygen and glucose deprivation/reoxygenation. In vivo experiments revealed that MEG3 knockdown suppressed autophagy, infarct volume and behavioral deficits in cerebral ischemia/reperfusion mice. These findings suggest that MEG3 knockdown inhibited autophagy and alleviated cerebral ischemia/reperfusion injury through the miR-181c-5p/ATG7 signaling pathway. Therefore, MEG3 can be considered as an intervention target for the treatment of cerebral ischemia/reperfusion injury. This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Zhengzhou University, China (approval No. XF20190538) on January 4, 2019.

[Clinical features and microsurgical management of rare tumors in the sellar region].

OBJECTIVE: To investigate the clinical characteristics and microsurgical managements of rare tumors in the sellar region. METHODS: Six rare cases of tumors in the sellar region treated by microsurgery from Jan 2000 to Jan 2010 were reviewed retrospectively. Subsequent treatments were according to the status of preoperative alpha fetal protein (AFP) and human chorionic gonadotropin (HCG) measurement as well as confirmed by histopathological examination in all six patients. RESULTS: Total resection of the tumor was achieved in 2 cases and subtotal resection in 4 cases. Postoperative histopathology confirmed that the lesions were tumors in 5 cases and fungal pseudotumor in 1 case. Moreover, variety of histological types were observed in the present series, including leiomyosarcoma, malignant yolk sac tumor, mixed germ cell tumor, embryonal carcinoma, pilocytic astrocytoma and fungal pseudotumor, respectively. The serum levels of AFP and HCG were elevated to some extent in the patients with malignant yolk sac tumor, mixed germ cell tumor or embryonal carcinoma. Follow-up was conducted in all patients for 1 month to 3 years. The patients with malignant yolk sac tumor and embryonal carcinoma as well as leiomyosarcoma died in 5, 6, 10 months after operation, respectively. Subarachnoid hemorrhage occurred in the case of fungal pseudotumor at 2 months after surgery. The other two patients were surviving well. CONCLUSIONS: Rare non-germinomatous malignant germ cell tumors are predominantly susceptible to the sellar region. Furthermore, High misdiagnosis rate and poor prognosis are characteristic in the present study. Dynamic AFP and HCG detection may play an important role in the diagnosis of those non-germinomatous malignant germ cell tumors located in the sellar region. The importance of awareness of the presence of such rare lesions in the sellar region is emphasized.

Serum D-dimer as a potential new biomarker for prognosis in patients with thrombotic thrombocytopenic purpura.

Thrombotic thrombocytopenic purpura (TTP) is a life-threatening disease, and its mortality rate is 10% to 20%. However, there are currently only a few markers to predict the prognosis in patients with TTP. We aimed to identify several clinical indices and laboratory parameters for predicting the prognosis of TTP at admission.A single-centre observational cohort study that included patients with TTP from the First Affiliated Hospital of Zhengzhou University in China was conducted from January 1, 2012 to November 30, 2018. The primary outcome was prognosis, including in-hospital mortality, major thromboembolic events, or failure to achieve remission at discharge. We used the random forest method to identify the best set of predictors.Eighty-seven patients with TTP were identified, of whom 12 died during the treatment. The total number of patients within-hospital mortality, major thromboembolic events, and failure to achieve remission at discharge was 58. The machine learning method showed that the D-dimer level was the strongest predictor of the primary outcome. Receiver operating characteristic (ROC) analysis demonstrated that the sensitivity and specificity of the D-dimer level alone for identifying high-risk patients were 78% and 81%, respectively, with an optimum diagnostic cut-off value of 770 ng/mL. The area under the ROC curve (AUC) was 0.80, and the 95% confidence interval (CI) was 0.70 to 0.90.This study found that the D-dimer level exhibited a good predictive ability for prognosis in patients with TTP. These findings may aid in the development of new and intensive treatment strategies to achieve remission among high-risk patients. However, external validation is necessary to confirm the generalizability of our approach across populations and treatment practices.

[Treatment of brain injured rats through transplanting amniotic-derived mesenchymal stem cells in different ways].

OBJECTIVE: To compare the behavioral improvement to find the best transplantation approach for treating brain injury through transplanting amniotic-derived mesenchymal stem cells into brain injured rats in different ways. METHODS: Eighty brain injured Wista rats were randomly divided into a control group with brain injury alone (n=20) and a treatment group(n=60) which were further evenly divided into Group A (transplanted through the vena caudalis), Group B (transplanted through the ventriculus cerebri lateralis), and Group C (transplanted through the injured brain area). Each group was transplanted with amniotic-derived esenchymal stem cells, and their therapeutic efficacy would be evaluated through the neurological severity score (NSS). RESULTS: Compared with other groups, the behaviors of Group C had markedly improved. There was statistically significant difference in the 2 groups (P<0.05). Compared with the control group, the behaviors of Group A and Group B had marked improvement. There was statistically significant difference in the 3 groups (P<0.05). However, there was no significant difference between Group A and the control group (P>0.05). CONCLUSION: Transplanting the amniotic-derived mesenchymal stem cells into the injured brain area may be effective for brain injury in rats.

MicroRNA-21 promotes glioma cell proliferation and inhibits senescence and apoptosis by targeting SPRY1 via the PTEN/PI3K/AKT signaling pathway.

AIMS: Our study aims to investigate the effect of microRNA-21 (miR-21) on the proliferation, senescence, and apoptosis of glioma cells by targeting SPRY1 via the PTEN/PI3K/AKT signaling pathway. METHODS: Glioma tissues and brain tissues were collected for this study after surgical decompression for traumatic brain injury. RT-qPCR was employed to measure mRNA levels of miR-21, SPRY1, PTEN, PI3K, and AKT, and Western blotting was conducted to determine protein levels of SPRY1, PTEN, PI3K, AKT, p-AKT, Caspase-3, Caspase-9, P53, GSK3, and p-GSK3. Human glioma U87 cells were assigned into the blank, negative control (NC), miR-21 mimics, miR-21 inhibitors, siRNA-SPRY1, and miR-21 inhibitors + siRNA-SPRY1 groups, with human HEB cells serving as the normal group. Cell proliferation, cell cycle, and apoptosis were determined by MTT and flow cytometry, respectively. RESULTS: Compared with control group, an increased expression of miR-21, PI3K, AKT, p-AKT, P53, and p-GSK3, and a decreased expression of SPRY1, PTEN, Caspase-3, and Caspase-9 were observed in the glioma group, and no significant differences were found in the expression of GSK3. SPRY1 was verified to be the target gene of miR-21. Compared with the blank and NC groups, levels of PI3K, AKT, p-AKT, P53, and p-GSK3 increased while levels of SPRY1, PTEN, Caspase-3, and Caspase-9 decreased in the miR-21 mimics and siRNA-SPRY1 groups; the miR-21 inhibitors group reversed the tendency; furthermore, the miR-21 inhibitors group showed decreased cell proliferation but promoted apoptosis, which were opposite to the results of the miR-21 mimics and siRNA-SPRY1 groups. CONCLUSION: MicroRNA-21 might promote cell proliferation and inhibit cell senescence and apoptosis of human glioma cells by targeting SPRY1 via the PTEN/PI3K/AKT signaling pathway.

Influence of different surgical timing on outcome of patients with aneurysmal subarachnoid hemorrhage and the surgical techniques during early surgery for ruptured intracranial aneurysms.

AIM: This study aims to investigate the influence of different surgical timing on outcome of patients with aneurysmal subarachnoid hemorrhage and to explore the surgical techniques for ruptured intracranial aneurysms. MATERIAL AND METHODS: The clinical data were from 327 cases. 304 cases of the surgical group were further assigned to early surgery (89 cases), intermediate surgery(164 cases) and delayed surgery(51 cases) according to the surgical timing. The other 23 cases were the nonsurgical group. The ultimate outcome of all cases was graded according to the Glasgow Outcome Scale. After the cases of the no-surgical group were re-assigned to different surgical subgroups according to the rebleeding time, the ultimate outcome was graded once more. RESULTS: There was no significant difference among the 3 groups' pre-operative clinical data. After re-assigning the cases of no-surgical group to the different surgical subgroups, there was no significant difference among the 3 groups' preooperative clinical data, while the ultimate outcome grades of early surgery (3.6 ± 1.8) and intermediate surgery (3.5 ± 2.2) were superior to that of delayed surgery (2.9 ± 2.8). CONCLUSION: This retrospective study has demonstrated that early surgery can not only prevent re-rupture of aneurysm to decrease mortality rate but also improve the ultimate outcome.

Local thrombolysis for patients of severe cerebral venous sinus thrombosis during puerperium.

OBJECTIVE: To explore and evaluate the efficacy of intrasinus thrombolysis (IST) in patients with cerebral venous sinus thrombosis (CVST) during postpartum period. METHODS: 11 patients during postpartum period with CVST who received IST during July 2007-November 2011 were included. Urokinase was infused into the sinuses via a microcatheter. Magnetic resonance venography (MRV) was performed to assess the recanalization of venous sinuses. RESULTS: Before discharge, the intracranial pressure in 11 patients was under 200 mmH(2)O. MRV confirmed that venous sinus of 9 patients were smooth. The cortex venous and deep venous recovered to normal. Venous sinus of 2 patients recanalized partly, and cortex venous and deep venous had compensation. 9 patients had good outcome and 2 patients had only mild deficits. CONCLUSION: Intrasinus thrombolysis is safe and effective in patients with severe cerebral venous sinus thrombosis during postpartum period.

Isoliquiritigenin inhibits proliferation and induces apoptosis of U87 human glioma cells in vitro.

Isoliquiritigenin (ISL), a member of the flavonoids, has been demonstrated to possess antitumor activity in various cancer cell lines in vitro and in vivo. In this study, we investigated the antitumor effects of ISL on U87 glioma cells in vitro. As determined by MTT assay, ISL inhibited the proliferation of U87 cells in a time-dependent and dose-dependent manner. The results of fluorescence-activated cell sorting (FACS) analysis suggested that ISL induced the apoptosis of the U87 cells and blocked cell cycle progression at the S and G2/M phases. Moreover, it was identified that ISL induced the apoptosis of the U87 cells in a caspase-dependent manner. Although treatment with the pan-caspase inhibitor Z-VAD-FMK efficiently blocked the ISL-induced caspase activation, it did not eliminate the ISL-induced cell death. Further examination using western blot analysis revealed that ISL upregulated p21/WAF1 and p27. These results indicate that cell cycle arrest and the caspase-mediated apoptosis pathway may participate in the antiproliferative activity of ISL in U87 cells by regulating the expression of specific molecules.