NLRP10 ablation protects against ischemia/reperfusion-associated brain injury by suppression of neuroinflammation.

Ischemic stroke leads to neuronal cell death and induces a cascade of inflammatory signals that results in secondary brain damage. Although constant efforts to develop therapeutic strategies and to reveal the molecular mechanism resulting in the physiopathology of this disease, much still remains unclear. Membrane-bound Toll-like receptors (TLRs) and cytosolic nucleotide binding oligomerization domain (NOD)-like receptors (NLRs) are two major families of pattern recognition receptors that initiate pro-inflammatory signaling pathways. In the present study, we explored the role of NLRP10 in regulating inflammatory responses in acute ischemic stroke using the wild type (WT) and NLRP10 knockout (KO) mice by inducing middle cerebral artery occlusion/reperfusion (MCAO) injuries. The study first showed that NLRP10 was over-expressed in the ischemic penumbra of WT mice. Then, the brain infarct volume was significantly decreased, and the moving activity was improved post-MCAO in mice with NLRP10 knockout. Apoptosis was also alleviated by NLRP10-knockout, as evidenced by the decreased number of TUNEL-staining cells. Further, NLRP10 deficiency attenuated the activation of glia cells in hippocampus of mice with MCAO operation. NLRP10 inhibition ameliorated the levels of inflammatory factors in peripheral blood serum and hippocampus of mice after stroke. The activation of toll-like receptor (TLR)-4/nuclear factor-κB (NF-κB) signaling pathways was markedly suppressed by NLRP10 ablation in mice after MCAO treatment. Importantly, inflammasome, including NLRP12, ASC and Caspase-1, induced by MCAO in hippocampus of mice was clearly impeded by the loss of NLRP10. The results above were mainly verified in LPS-incubated astrocytes in the absence of NLRP10. Correspondingly, in LPS-treated astrocytes, NLRP10 knockout-reduced inflammation via impairing TLR-4/NF-κB and NLRP12/ASC/Caspase-1 pathways was evidently restored by over-expressing NLRP10. Therefore, the results above indicated an essential role of NLRP10 in regulating ischemic stroke, presenting NLRP10 as a promising target to protect human against stroke.

11 Long noncoding RNA UCA1 functions as miR-135a sponge to promote the epithelial to mesenchymal transition in glioma.

The dysregulation of long noncoding (lncRNA) UCA1 may play an important role in tumor progression. However, the function in gliomas is unclear. Therefore, this experiment was designed to explore the pathogenesis of glioma based on lncRNA UCA1. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of lncRNA UCA1, miR-135a, and HOXD9 in gliomas tissues. The effect of lncRNA UCA1 and miR-135a on tumor cell proliferation and migration invasiveness was examined by CCK-8 and transwell assays. Target gene prediction and screening, luciferase reporter assay were used to verify downstream target genes of lncRNA UCA1. Expression of E-cadherin, N-cadherin, vimentin, and HOXD9 was detected by RT-qPCR and Western blotting. The tumor changes in mice were detected by in vivo experiments in nude mice. lncRNA UCA1 was highly expressed in glioma tissues and cell lines. lncRNA UCA1 expression was associated with significantly poor overall survival in gliomas. Moreover, lncRNA UCA1 significantly enhanced cell proliferation and migration, and promoted the occurrence of EMT. In addition, lncRNA UCA1 promoted the development of EMT by positively regulating HOXD9 expression as a miR-135a sponge. In vivo experiments indicated that UCA1 exerted its biological functions by modulating miR-135a and HOXD9. In conclusion, lncRNA UCA1 can induce the activation of HOXD9 by inhibiting the expression of miR-135a and promote the occurrence of EMT in glioma.

Application of Y-shaped, coated self-expandable metallic stents for anastomotic stenosis after gastrojejunostomy (Billroth II).

BACKGROUND: Anastomotic stenosis is an infrequent but life-threatening complication after gastrojejunostomy (Billroth II). Tubular or single tubular stents have limited efficacy due to the particular anatomy. PURPOSE: To assess the feasibility of a Y-shaped, fully-coated, self-expandable, metallic stent (SEMS) for anastomotic stenosis after gastrojejunostomy (Billroth II). MATERIAL AND METHODS: Between January 2008 and August 2014, 14 patients (10 with gastric carcinoma and four with duodenal ulcers) had anastomotic stenoses following Billroth II reconstructions. Eight patients with gastric cancer had tumor recurrence near the anastomosis; two had benign strictures. The four duodenal ulcer patients had benign stenoses. An integrated Y-shaped, fully coated SEMS was designed to accord with the anatomy of residual gastrojejunal anastomotic strictures. Fourteen stents were inserted under fluoroscopic control. Follow-up was at 1, 3, 9, and 12 months, and then annually. RESULTS: All 14 stents were inserted successfully at the first attempt with a technical success rate of 100%. After stenting, abdominal symptoms resolved in all patients. All patients were followed up for 4-27 months (mean, 13.9 months). One of the eight recurrent cases died of multiple tumor metastases and liver failure after 7 months, without obstruction symptoms. In all six patients with benign anastomotic stenosis, the stents were removed successfully without complication and with no evidence of restenosis based on clinical evaluation and imaging. CONCLUSION: A Y-shaped, fully-coated SEMS proved to be a feasible and minimally invasive procedure for treating anastomotic stenosis after gastrojejunostomy (Billroth II).

Value of percutaneous transhepatic cholangiobiopsy for pathologic diagnosis of obstructive jaundice: analysis of 826 cases.

BACKGROUND: Obstructive jaundice (OJ) is insensitive to radiation and chemotherapy, and a pathologic diagnosis is difficult to make clinically. Percutaneous transhepatic cholangiobiopsy (PTCB) is simple to perform and minimally invasive, and clinical practice has shown it to be an accurate and reliable new method for bile duct histopathologic diagnosis. PURPOSE: To investigate the value of PTCB for pathologic diagnosis of causes of OJ. MATERIAL AND METHODS: From April 2001 to December 2011, PTCB was performed in 826 consecutive patients. Data on pathologic diagnosis, true positive rate, and complications were analyzed retrospectively. Patients with negative pathologic findings were diagnosed using clinical, imaging, laboratory, and prognostic data. The feasibility and safety of PTCB for OJ were evaluated and true positive rates for biliary carcinoma and non-biliary carcinoma compared. RESULTS: PTCB was successful in all cases. Of 740 patients clinically diagnosed with malignant biliary stricture and 86 with benign biliary stricture, 727 received a positive pathologic diagnosis; in 99, the pathologic findings were considered false negative. The true positive rate for PTCB was 88.01% overall, differing significantly for biliary and non-biliary carcinoma (χ2 = 12.87, P < 0.05). Malignancy accounted for 89.59% of OJ cases; well, moderately, and poorly differentiated carcinoma represented 57.88%, 19.97%, and 22.15%. Biliary adenocarcinoma was the predominant malignant pathologic type (96.41%). Transient bilemia, bile leakage, and temporary hemobilia occurred in 47, 11, and 28 cases, respectively, with no serious complications. CONCLUSION: PTCB is safe, feasible, and simple, with a high true positive rate for definitive diagnosis of OJ causes. Well differentiated adenocarcinoma was the predominant pathologic type.

Balloon dilatation and thrombus extraction for the treatment of cerebral venous sinus thrombosis.

BACKGROUND AND PURPOSE: This study aimed to investigate the efficacy and safety of balloon dilatation and thrombus extraction for the treatment of cerebral venous sinus thrombosis (CVST). MATERIALS AND METHODS: Twenty-six cases of digital subtraction angiography-confirmed CVST were treated with balloon dilatation and thrombus extraction. Active treatment of primary disease was carried out after cerebral venous sinus recanalization, and the subsequent anticoagulant therapy lasted for 6 months. RESULTS: Recanalization of the cerebral venous sinus was achieved in all 26 patients, and no endovascular treatment-related complications occurred during or after the procedure. At discharge, the Glasgow Coma Scale (GCS) of the patients had improved from an average of 12.3 points to 15 points, and clinical symptoms were improved in 100% of the patients. Follow-up times ranged from 12-62 months (mean follow-up time of 42.3 months) and no thrombus re-formation or new neurological deficits occurred during that time. CONCLUSION: Based on our small study population, balloon dilatation and thrombus extraction appears to be a safe and effective treatment for cerebral venous sinus thrombosis. However, further research is needed to confirm this.

IL-21R functions as an oncogenic factor and is regulated by the lncRNA MALAT1/miR-125a-3p axis in gastric cancer.

Interleukin-21 receptor (IL-21R) is involved in the immunological regulation of immune cells and tumor progression in multiple malignancies. However, the potential molecular mechanisms through which non-coding RNAs (ncRNAs) modulate IL-21R signaling in gastric cancer (GC) remain elusive. In this study, the expression of IL-21R was detected by RT-qPCR and western blot analysis in GC cell lines. The association between IL-21R expression and clinicopathological characteristics and the prognosis of patients with GC was analyzed by immunohistochemistry and Kaplan-Meier plotter analysis. The biological functions of IL-21R were analyzed by a series of in vitro and in vivo experiments, and its regulation by ncRNAs was predicted by bioinformatics analysis and confirmed by luciferase assays and rescue experiments. As a result, the expression of IL-21R was found to be significantly increased in GC cell lines and tissues as compared with normal tissues, and was associated with tumor size and lymphatic metastasis, acting as an independent prognostic factor of poor survival and recurrence in patients with GC. The knockdown of IL-21R markedly suppressed GC cell proliferation and invasion, and IL-21R expression was further validated to be negatively regulated by miR-125a-3p (miR-125a). The overexpression of IL-21R reversed the tumor suppressive effects of miR-125a in vitro and in vivo. Moreover, lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acted as a sponge of miR-125a to modulate the IL-21R signaling pathway in GC cells and represented a risk factor for survival and recurrence in patients with GC. Taken together, the findings of this study reveal an oncogenic role for IL-21R in gastric tumorigenesis and verify that its activation is partly due to the dysregulation of the lncRNA MALAT1/miR-125a axis. These findings may provide a potential prognostic marker for patients with GC.

Chrysin ameliorates cerebral ischemia/reperfusion (I/R) injury in rats by regulating the PI3K/Akt/mTOR pathway.

In this study, the effects of chrysin on cerebral ischemia by establishing middle cerebral artery occlusion (MCAO) in rat were investigated. In vivo experiments, the rats were orally administrated with clopidogrel or chrysin once daily for 7 days before the experimental of ischemia and the rats were divided into 5 groups: the sham group, the I/R group, I/R + clopidogrel group, I/R + chrysin (10 mg/kg), I/R + chrysin (20 mg/kg) group. Chrysin significantly ameliorated the I/R rats, evaluated by TTC staining, determination of brain wet to dry weight ratio and neurological deficits. Moreover, in serum and brain tissues of the I/R rats, chrysin also could effectively suppress the release of inflammatory cytokines, including levels of interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α). In addition, chrysin could improve the SOD activity in the I/R rats. Mechanically, chrysin could activate the PI3K/Akt/mTOR pathway, inhibited inflammation and apoptosis. In oxygen-glucose deprivation and recovery (OGD/R)-induced SH-SY5Y cells in vitro. Chrysin markedly decreased the levels of TNF-α, IL-6 and IL-1ß in supernatant of OGD/R-induced SH-SY5Y cells via activating PI3K/Akt/mTOR pathway. In conclusion, our study demonstrated that chrysin might be a potential therapeutic agent for cerebral ischemia.

One-Stage Endovascular Embolization for Multiple Intracranial Aneurysms.

AIM: Clinical treatment of multiple intracranial aneurysms remains challenging due its higher rate of rupture compared to a single aneurysm. We aimed to assess the efficacy of one-stage endovascular embolization for treatment of multiple intracranial aneurysms. MATERIAL AND METHODS: We treated 72 aneurysms from 33 patients with one-stage endovascular embolization from September 2010 to October 2015. Of these, 60 aneurysms were treated with coils, while 12 wide-neck aneurysms were embolized with the assistance of intracranial stents. Follow-up studies ranged from 3 to 28 months, and patient outcomes were assessed using the modified Rankin Scale (mRS) and digital subtraction angiography (DSA). RESULTS: All aneurysms were embolized successfully and no complications occurred. DSA immediately after procedure revealed that complete occlusion was achieved in 39 cases, neck remnant was observed in 27 cases, and a residual aneurysm remnant was observed in six cases. All patients achieved excellent clinical outcomes following embolization (27 patients with a mRS score of 0 and 6 with a mRS score of 1). Six months after the procedure, DSA was performed on 28 patients, revealing one case with a neck remnant, and two cases with completely occluded aneurysms. Despite these outcomes, there was no rupture or hemorrhage of these aneurysms throughout the six months. CONCLUSION: One-stage endovascular embolization is a safe and effective treatment for multiple intracranial aneurysms, but long-term outcomes should be further evaluated.

Dysregulation of microRNA­23b­3p contributes to the development of intracranial aneurysms by targeting phosphatase and tensin homolog.

MicroRNA­23b­3p (miR­23b­3p) has been reported to be involved in the pathogenesis of a number of diseases, including non­small cell lung cancer and gastric cancer, by acting on different signaling pathways. The present study aimed to understand the association between the miR­23b­3p level of intracranial aneurysms (IAs) and the mechanism involved. Computational analysis was used to search for the target of miR­23b­3p, and luciferase assay was used to validate the miRNA/target association. Western blot analysis and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to determine the expression of miR­23b­3p and phosphatase and tensin homolog (PTEN), and their expression in smooth muscle cells (SMCs) treated with miRNA mimic or inhibitor. Firstly, an online miRNA database (www.mirdb.org) was searched using the 'seed sequence' located within the 3'­untranslated region of the target gene, and then PTEN was validated as the direct target gene via a luciferase reporter assay system. The negative regulatory association between miR­23b­3p and PTEN was determined through the analysis of the relative luciferase activity. Additionally, RT-qPCR and western blot analysis was performed in order to assess the mRNA and protein expression levels of PTEN among IA (n=32) and control (n=17) groups or cells treated with scramble control, miR­23b­3p mimics, PTEN siRNA and miR­23b­3p inhibitors to verify the negative regulatory association between miR­23b­3p and PTEN. Experiments were then performed to investigate the effect of miR­23b­3p and PTEN on the viability and apoptosis of pulmonary artery SMCs (PASMCs). The results showed that cells transfected with miR­23b­3p inhibitors suppressed the viability of SMCs by promoting the apoptosis of the cells compared with that of the scramble controls, while cells transfected with miR­23b­3p mimics and PTEN siRNA enhanced the viability of VSMCs by inducing apoptosis. This indicated that miR­23b­3p negatively interfered with the viability of the cells, while PTEN positively interfered with the viability of the cells. In conclusion, PTEN was found to be a virtual target of miR­23b­3p, and a negative regulatory association existed between miR­23b­3p and PTEN. miR­23b­3p and PTEN interfered with the viability and apoptosis of SMCs.

Study on the role of Cathepsin B and JNK signaling pathway in the development of cerebral aneurysm.

OBJECTIVE: To investigate the correlation between JNK signal and the apoptosis of VSMC as well as the expression of Cathepsin B and to explore the role of JNK signal in the development of cerebral aneurysm. METHODS: Rat models of cerebral aneurysm were established and histopathologic changes of cerebral aneurysm and the apoptosis of VSMC were analyzed. Rat models were respectively subject to subcutaneous injection of Cathepsin B siRNA and JNK inhibitor SP600125. Western blot technique was used to detect the expression of proteins like Cathepsin B, Caspase-3, and p-JNK. Spearman's rho was used to examine the correlation between p-JNK and Cathepsin B, as well as the expression of relevant proteins. RESULTS: The success rate of modeling rats with cerebral aneurysm was 88.75%. After the respective injection of Cathepsin B siRNA, SP600125 and their combination, the cell densities of VSMC of rats with cerebral aneurysm all increased significantly (P < 0.05 or P < 0.01), but the apoptosis rate of VSMC decreased significantly (P < 0.01). Compared with normal rats, the expression of Cathepsin B, Caspase-3 and p-JNK in Cerebral aneurysm models increased significantly. Effectively intervening Cathepsin B genes with Cathepsin B siRNA could significantly inhibit the expression of Cathepsin B and Caspase-3, but hardly influence the expression of p-JNK. JNK inhibitor SP600125 had no influence on the expression of Cathepsin B and Caspase-3, but effectively inhibited the expression of p-JNK. In cerebral aneurysm tissues, positive correlation was observed between the expression of p-JNK and Cathepsin B, the correlation coefficient was r = 0.640. CONCLUSION: After the attack of cerebral aneurysm, proteins like Cathepsin B, Caspase-3 and p-JNK are all involved in the apoptosis of VSMCs. This process may be realized by Cathepsin B which activates the apoptosis mechanism of Caspase-3 and mediate the apoptosis of VSMC through the JNK signaling pathway. Therefore, silencing Cathepsin B gene or inhibiting the conduction through JNK signaling pathway can mitigate the apoptosis of vascular smooth muscle cells in cerebral aneurysm.

Stent-assisted mechanical recanalization for symptomatic subacute or chronic middle cerebral artery occlusion.

To assess the feasibility and short-term effects of treating patients with subacute or chronic middle cerebral artery (M1) occlusion by stent-assisted mechanical recanalization. Six patients with cerebral arteries occlusion underwent surgery. Six cerebral arteries occlusion in 5 patients were successfully recanalized. On postoperative day 1, four patients' symptoms were relieved and two patients' symptoms were exacerbated, of which one was significantly improved after 3 days, the other one's symptoms were recovered to preoperative levels in 2 weeks. No patients died after surgery. No stroke or transient ischemic attack occurred. The average follow-up of was 4.2 months, no worsening of condition, recurrence or death occurred. The results indicate that for patients with subacute or chronic middle cerebral artery (M1) occlusion, mechanical recanalization was technically feasible under the premise of strict case screening. Mechanical recanalization is able to improve ischemic symptoms and promote dysfunction restoration. But its long-term effect remains to be evaluated by further large samples, long-term follow-up studies.

The Use of Solitaire AB Stents in Coil Embolization of Wide-Necked Cerebral Aneurysms.

BACKGROUND: The Solitaire AB stent is one of many assistant stents used for treating wide-necked cerebral aneurysm, and has been used since 2003. However, large sample studies on its safety and effectiveness are lacking. The objective of this study was to evaluate the effectiveness and safety of the Solitaire AB stent in the coil embolization of wide-necked cerebral aneurysms. METHODS: Retrospective review of the clinical and image data of 116 patients with wide-necked cerebral aneurysms who had been enrolled at six interventional neuroradiology centers from February 2010 to February 2014 and had been treated by coil embolization; in total, 120 Solitaire AB stents were used. The degree of aneurysm occlusion was examined using digital subtraction angiography (DSA) immediately after the procedure and during follow-up, and was graded using the modified Raymond classification. We also observed complications to evaluate the safety and effectiveness of this therapy. RESULTS: The 120 Solitaire AB stents (4 mm × 15 mm, four stents; 4 mm × 20 mm, 16 stents; 6 mm × 20 mm, 36 stents; 6 mm × 30 mm, 64 stents) were inserted to treat 120 wide-necked cerebral aneurysms. All stents were inserted successfully. DSA immediately post-surgery revealed 55 cases of complete occlusion, 59 cases of neck remnant, and six cases of aneurysm remnant. Perioperatively, there were four cases of hemorrhage and four cases of stent thrombosis. The follow-up spanned 3-37 months; of 92 patients examined by DSA at the 6-month follow up, 12 had disease recurrence. CONCLUSIONS: The Solitaire AB stent is effective with a good technical success rate and short-term effect for assisting coil embolization of wide-necked cerebral aneurysms.