Rescuing defective tumor-infiltrating T-cell proliferation in glioblastoma patients.

Primary glioblastoma (GBM) is the most prevalent brain cancer, with fast progression and a poor prognosis. Current treatment options are unable to fully manage GBM since it is highly resistant to radiation and chemotherapy, and it cannot be completely removed by surgery. Thus, immunotherapeutic strategies utilizing tumor-infiltrating T cells have been investigated. In the present study, the T-cell response in GBM patients was examined in resected tumor samples and peripheral blood samples by flow cytometry. It was found that tumor-infiltrating T cells represented a rare population in all tumor cells, and were more refractory to anti-cluster of differentiation 3 (CD3) stimulation than their peripheral blood counterparts. A number of strategies were then assessed to boost tumor-infiltrating T-cell proliferation, and it was found that pre-incubation with 20 U/ml interleukin (IL)-2, as well as sequestration of IL-10 in culture, improved tumor T-cell proliferation following anti-CD3 stimulation. The stimulation of blood antigen-presenting cells by lipopolysaccharide, however, did not improve tumor T-cell proliferation. Overall, the present results provided a viable strategy for improving tumor-infiltrating CD3+ T-cell responses in GBM patients.

The Expression Levels of XLF and Mutant P53 Are Inversely Correlated in Head and Neck Cancer Cells.

XRCC4-like factor (XLF), also known as Cernunnos, is a protein encoded by the human NHEJ1 gene and an important repair factor for DNA double-strand breaks. In this study, we have found that XLF is over-expressed in HPV(+) versus HPV(-) head and neck squamous cell carcinoma (HNSCC) and significantly down-regulated in the HNSCC cell lines expressing high level of mutant p53 protein versus those cell lines harboring wild-type TP53 gene with low p53 protein expression. We have also demonstrated that Werner syndrome protein (WRN), a member of the NHEJ repair pathway, binds to both mutant p53 protein and NHEJ1 gene promoter, and siRNA knockdown of WRN leads to the inhibition of XLF expression in the HNSCC cells. Collectively, these findings suggest that WRN and p53 are involved in the regulation of XLF expression and the activity of WRN might be affected by mutant p53 protein in the HNSCC cells with aberrant TP53 gene mutations, due to the interaction of mutant p53 with WRN. As a result, the expression of XLF in these cancer cells is significantly suppressed. Our study also suggests that XLF is over-expressed in HPV(+) HNSCC with low expression of wild type p53, and might serve as a potential biomarker for HPV(+) HNSCC. Further studies are warranted to investigate the mechanisms underlying the interactive role of WRN and XLF in NHEJ repair pathway.

Recombinant adenoviral vector expressing human wild-type p53, GM-CSF, and B7-1 genes suppresses the growth of glioma in vivo.

Malignant gliomas are the most common of primary brain tumors and have been proven incurable with conventional treatments. Evidence have shown that a recombinant adenoviral vector expressing human wild-type p53, granulocyte-macrophage colony-stimulating factor (GM-CSF), and B7-1 genes (BB-102) may have antitumor effects in vitro. In this study, we investigated the effects of BB-102-based vaccine on glioma in vivo. An animal model using nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice with human immune system was established. The mice were vaccinated with inactivated U251 glioma cells transduced with BB-102 or adenoviral vector expressing green fluorescence protein (Ad-GFP) as a control and followed by the challenge of live U251 glioma cells. Tumor growth and antitumor responses were measured. Data showed that mice vaccinated with BB-102 had significantly reduced local tumor growth compared to mice with Ad-GFP vaccination or the control group. Histopathological analysis displayed low tumor cell density and significant infiltration of human peripheral blood lymphocytes (HuPBLs) in the tumor tissues of mice transduced with BB-102. Immunohistochemical analysis showed that mutant p53 was not expressed in tumor tissues of mice with BB-102 vaccination, and the expression level of Ki67 was significantly lower in the tumor tissues of the BB-102 group than those in the Ad-GFP group or the control group. Further study demonstrated that mice with BB-102 vaccination had significantly increased total T cell numbers, total T cell proportion, CD4+ T cell proportion, and CD8+ T cell proportion in spleens, as well as higher value of IgG, IgA, and IgE in sera. These data suggest that the recombinant adenoviral vector expressing human wild-type p53, GM-CSF, and B7-1 genes could suppress glioma in NOD/SCID mice model and might be considered as a novel strategy for glioma therapy.

Lysyl oxidase genetic variants and the prognosis of glioma.

Lysyl oxidase (LOX) is a copper-dependent amine oxidase that plays important roles in the development and homeostasis of primary brain tumors such as glioma. The aim of this study was to investigate whether polymorphisms in the LOX gene were associated with susceptibility to glioma. We tested two functional polymorphisms of LOX, -22G/C and 473G/A, and compared them between 466 glioma cases and 502 healthy controls in the Chinese population. Results showed that the prevalence of 473AA genotype was significantly increased in cases than in controls (p = 0.001). Individuals who carried 473A allele had a 1.44-fold of increased risk for glioma than those with 473G allele (p = 0.002). In addition, when analyzing the survival time of glioma patients with LOX 473G/A polymorphism, cases with AA genotype had significantly shorter survival time compared to the patients carrying G allele (25.0 months vs 43.0 months, p = 0.0009). These results suggested that polymorphism in LOX gene was associated with increased susceptibility to glioma and could be used as prognostic factor for this malignancy.

[Surgical treatment of thoracic tuberculosis with one stage posterior debridement and bone grafting fusion and internal fixation].

OBJECTIVE: To investigate the effect and indication of one stage posterior debridement and bone grafting fusion and internal fixation for thoracic tuberculosis. METHODS: From January 2005 to May 2011,12 patients with thoracic tuberculosis were treated with one stage posterior debridement and pedicle screw fixation combined with regular anti-tuberculosis treatment before and after operation. There were 7 males and 5 females,with an average age of 45 years and average course of 15 months. Information of operative time, blood loss, bony fusion, local kyphosis and neurologic functional were evaluated. RESULTS: All infective focus were thoroughly removed and bone graft obtained fusion. The mean of operative time and blood loss were 170 min (120-210 min) and 510 ml (200-1 000 ml),respectively. Cobb angle from (28.7 +/- 9.2) degrees preoperatively decreased to (8.2 +/- 3.5) degrees postoperatively(P<0.05). No kyphosis correction loss,tubercular recurrence or failure of internal fixation was found. According to Frankel grade to evaluate neurological function, all patients arrived to grade E. CONCLUSION: One stage posterior debridement and bone grafting fusion and internal fixation is an effective method in treating thoracic tuberculosis. It has advantages such as thorough debridement, short operative time, less blood loss, more kyphosis correction and higher bony fusion rate.

Wingspan stent-assisted coiling of intracranial aneurysms with symptomatic parent artery stenosis: experience in 35 patients with mid-term follow-up results.

BACKGROUND: There is a potential risk of aneurysm rupture after parent artery revascularization because of increased blood flow. The purpose of this study is to assess the efficacy and safety of Wingspan stent-assisted coil embolization in the treatment of intracranial aneurysms with symptomatic parent artery stenosis. METHODS: Thirty-five consecutive patients (19 men, 16 women; age range, 48-79 years; mean age, 60.4 years) harboring 35 unruptured wide-necked or fusiform intracranial aneurysms (mean size 6.8mm; range 2.5-18 mm.) with symptomatic parent artery stenosis (mean degree 71.1%; range 50-92%) were treated with the Wingspan stent-assisted coiling. Twenty-four lesions were located in the anterior circulation and eleven in the posterior circulation. Patients were premedicated with antiplatelet therapy consisting of aspirin 300 mg and clopidogrel 75 mg for at least 3 days before the procedure. Following pre-dilatation and stent placement, a coiling microcatheter entered the aneurysm through the interstices of the stent, and then coiling was performed. After the procedure, clopidogrel 75 mg daily was recommended for an additional 30 days, and aspirin 100mg was recommended throughout follow-up. For all patients, clinical follow-up was conducted by clinic visitation, or telephone interview. Angiographic follow-up with DSA was recommended at 6 months and 1 year after the procedure. Angiography follow-up (mean time 10.6 months) was obtained in 31 cases (88.6%). The technical feasibility of the procedure, procedure-related complications, angiographic results, clinical outcome and follow-up angiography were evaluated. RESULTS: In every case, technical success was achieved. The degree of stenosis was reduced from 71.1% to 17.4% after balloon angioplasty and stenting. Immediate angiography demonstrated complete occlusion in 25 cases (71.4%), neck remnant in 7 cases (20.0%), and incomplete occlusion in 3 cases (8.6%). Procedure-related morbidity occurred in two patients (5.7%), including thromboembolism (n=1) and occlusion of small penetrating arteries (n=1). At follow-up (mean time 18.3 months), two additional cases of ischemic stroke occurred. The overall frequency of any stroke, intracranial hemorrhage, or death within 30 days or ipsilateral stroke beyond 30 days was 11.4%. No rehemorrhage of treated aneurysm occurred. At angiographic follow-up, four cases demonstrated ≥ 50% in-stent restenosis (12.9%), one of which was symptomatic, and two aneurysms (6.4% of the follow-up angiograms) demonstrated recanalization. CONCLUSION: We found that the Wingspan stent-assisted coil embolization was helpful in the treatment of intracranial aneurysms with parent artery stenosis.

Intra-aneurysmal Onyx embolization for distal aneurysms of the cerebellar arteries.

BACKGROUND: Distal cerebellar artery aneurysms are difficult to treat both surgically and endovascularly. The purpose of this study is to assess the efficacy and safety of intra-aneurysmal Onyx embolization of these lesions with parent artery preservation. METHODS: Ten consecutive patients harboring 10 distal cerebellar aneurysms were treated with intra-aneurysmal Onyx embolization. Locations of the aneurysms were as follows: superior cerebellar artery in three patients, anterior inferior cerebellar artery in two, and posterior inferior cerebellar artery in five. The technical feasibility of the procedure, procedure-related complications, angiographic results, and clinical outcome were evaluated. RESULTS: In every case, endovascular treatment was achieved. Immediate angiography demonstrated that all of 10 aneurysms were completely occluded, with parent artery preservation in nine of them. Procedure-related complication occurred in one patient with transient neurological sequelae. None of the treated aneurysms experienced rebleeding or recanalization during the follow-up time (mean, 12·7 months). CONCLUSION: We found that the intra-aneurysmal Onyx embolization was helpful in the treatment of distal cerebellar aneurysms in which selective occlusion of the aneurysmal sac with coils or open clipping cannot be achieved.

A single-centre experience and follow-up of patients with endovascular coiling of large and giant intracranial aneurysms with parent artery preservation.

Large and giant aneurysms are some of the most challenging vascular pathologies in the central nervous system. Their peculiarities make the surgical and endovascular approaches difficult and frequently limit them by posing risks and complications. Endovascular coil embolization of these lesions is being used increasingly as an alternative. Here we report the clinical experience and follow-up results of the endosaccular packing of 102 consecutive patients with 106 large or giant aneurysms to assess the efficacy and safety of this method. Embolization was completed by packing the aneurysm sac with a variety of commercially available coils. Primary endosaccular coiling, balloon-assisted coiling and stent-assisted coiling were used. The technical feasibility of the procedure, procedure-related complications, angiographic results, clinical outcome and follow-up angiography were evaluated. During admission, immediate angiography demonstrated complete occlusion in 48.1%, neck remnant in 28.3%, and incomplete occlusion in 23.6%. Procedure-related morbidity and mortality was 7.5% and 2.8%, respectively. A favorable clinical outcome (Modified Rankin Scale score of 0-2) was observed in 88.2% of patients (average follow-up time, 56.5 months). No re-hemorrhage of a treated aneurysm occurred. Angiography follow-up was obtained in 77.5% (79/102) patients (average follow-up time, 38.1 months). The overall recanalization rate was 29.6%. Comparison of occlusion class immediately after treatment and at last follow-up showed that 80.2% of the 81 aneurysms (in 79 patients) were stable or had improved. Five stent-assisted aneurysms that were not completely occluded initially had converted to complete occlusion on last follow-up. Nineteen recanalized aneurysms underwent successful re-embolization. No procedural complication was seen at retreatment. We conclude that in treating large and giant intracranial aneurysms, endovascular coiling with parent vessel preservation is a safe and effective technique.

Complications and adverse events associated with Neuroform stent-assisted coiling of wide-neck intracranial aneurysms.

BACKGROUND: Successful experiences of the Neuroform stent-assisted coiling have been reported by many teams in endovascular neurosurgery centers throughout the world. However, most of the reported complications involved a limited number of patients. OBJECTIVE: To systematically report the complications of Neuroform stent-assisted coiling of intracranial aneurysms and to tentatively assess the efficacy and safety of this method. METHODS: A retrospective study of 232 consecutive patients with 239 wide-neck aneurysms treated with Neuroform stent-assisted coil embolization at our institution over a 6-year period was performed. Angiographic results and clinical outcome were evaluated. Cases with complications were analyzed. RESULTS: Stenting was successful in 237 of 239 aneurysms. Favorable clinical outcome (modified Rankin score: 0-2) was observed in 88·3% of the patients. Procedure-related complications included thromboembolism (n = 13), intraprocedural rupture (n=8), coil protrusions (n=5), new mass effect (n=3), vessel injury (n=3), and stent dislodgement (n=2). Procedure-related morbidity and mortality were 4·2 and 1·3%, respectively. Non-procedural complications attributable to subarachnoid hemorrhage in 129 patients with ruptured aneurysms were symptomatic vasospasm (18·6%) and shunt-dependent hydrocephalus (6·9%). Angiography follow-up was obtained in 67·1% of the treated aneurysms. The overall recanalization rate was 14·5%. Delayed complications included in-stent stenosis (n=2) and penetrating artery occlusion (n=2) in follow-up period. CONCLUSION: Neuroform sent-assisted coiling of intracranial aneurysm is a safe technique with relatively low recanalization rate. The main cause of morbidity and mortality is thromboembolism. Long-term effect on parent artery should be observed carefully.

Endovascular treatment for dural arteriovenous fistulas at the jugular foramen.

We report the technique and results of the endovascular treatment of jugular foramen dural arteriovenous fistulas (DAVFs) in 4 (3 men and 1 women, mean age 50.75 years) symptomatic patients. The jugular foramen DAVFs accounted for 5.9% of intracranial DAVFs. Three patients presented with pulsatile tinnitus and 1 patient presented with intracranial hemorrhage. Angiography demonstrated an AV fistula at the jugular foramen, mostly arising from the middle meningeal, ascending pharyngeal and vertebral arteries with direct drainage to the internal jugular vein. All patients underwent transarterial embolization using Onyx-18. Complete shunt obliteration was achieved in 3 patients; and shunt reduction, in 1 patient, who was cured with additional surgery. Our study suggests that in jugular foramen DAVF transarterial embolization with Onyx should be considered when access is available.

Adenovirus mediated transfer of p53, GM-CSF and B7-1 suppresses growth and enhances immunogenicity of glioma cells.

OBJECTIVES: Malignant gliomas are good targets for gene therapy because they have been proven incurable with conventional treatments. However, malignant gliomas are genetically and physiologically highly heterogeneous, and current gene therapy interventions have been designed to target only a few variations of this kind of disease. Hence, we developed a combined gene therapy approach using a recombinant adenovirus carrying human wild-type p53 (WT-p53), granulocyte-macrophage colony-stimulating factor (GM-CSF) and B7-1 genes (designated BB-102) to combat the disease. METHODS: Human malignant glioma cells U251 and U87 were transduced with BB-102. Expression of WT-p53, GM-CSF and B7-1 genes were determined by Western blot, enzyme linked immunosorbent assay and flow cytometric analysis, respectively. Growth rates were determined by serial cell counts. Apoptosis was detected by flow cytometric analysis. Proliferation of autologous peripheral blood lymphocytes (PBLs) and cytotoxicity against primary glioma cells were assessed by cell proliferation and cytotoxicity assay kits, respectively. RESULTS: By the transduction of BB-102, high expression levels of the three exogenesis genes were detected in glioma cells. Cell growth was inhibited and apoptosis was induced. Significant proliferation of autologous PBLs and specific cytotoxicity against primary glioma cells were also induced by the infection of BB-102 in vitro, with the effect being more evident than that of Ad-p53. CONCLUSION: These results suggest that glioma cell vaccination co-transferred with p53, GM-CSF and B7-1 genes may be a feasible and effective immunotherapeutic approach in glioma treatments.

Neuroform stent-assisted coiling of intracranial aneurysms: a 5 year single-center experience and follow-up.

BACKGROUND AND PURPOSE: Wide-necked and fusiform aneurysms still remain a therapeutic challenge both surgically and endovascularly. The authors report their clinical experience and 5 year follow-up results using Neuroform stent, as an adjunct in the treatment of wide-necked and fusiform aneurysms. METHODS: One hundred and seven consecutive patients with 110 wide-necked or fusiform intracranial aneurysms were treated with the Neuroform stent-assisted coiling. Both sequential technique and parallel technique were used. In all cases, embolization was completed by packing the aneurysm sac with a variety of commercially available coils. The technical feasibility of the procedure, procedure-related complications, angiographic results, clinical outcome and follow-up angiography were evaluated. RESULTS: In every case, the Neuroform stent system was delivered and deployed accurately, and occlusion was achieved. Immediate angiography demonstrated complete occlusion in 57.2%, neck remnant in 27.3% and incomplete occlusion in 15.5%. Procedure-related morbidity was 5.6% and procedural-related mortality was 0.9%. Favorable clinical outcome (modified Rankin scale score: 0-2) was observed in 90.7% of the patients (average follow-up time: 47.3 months). No rehemorrhage of treated aneurysm occurred. Angiography follow-up was obtained in 45.8% (49/107 patients; 51/110 aneurysms; average follow-up time: 37.2 months). The overall recanalization rate was 13.7%. Comparison of occlusion class immediately after treatment and at last follow-up showed that 86.3% of the 51 aneurysms had stable or improved class. Eight aneurysms (36.4%) that were not initially completely occluded converted to complete occlusion on follow-up. No delayed coil or stent migration was found. One patient with in-stent stenosis and one with penetrating artery occlusion occurred as delayed complications. CONCLUSION: In treating complex intracranial aneurysms, the Neuroform stent-assisted coiling is a secure and effective technique.

Pivotal involvement of neurogenic mechanism in subcutaneous bee venom-induced inflammation and allodynia in unanesthetized conscious rats.

The bee venom (BV) model is a valid inflammatory pain model in animals and has been extended to human studies using its principle protein, mellitin. After subcutaneous (s.c.) injection of BV, long-lasting spontaneous nociception followed by thermal hyperalgesia, static allodynia, and local inflammatory response (edema) can be observed in rats. We hypothesize that (1) neurogenic components may contribute to the BV-induced inflammatory response and (2) static and dynamic mechanical allodynia may exist simultaneously in the BV model. Using different approaches including sciatic nerve transection (SCT), L4-L6 dorsal rhizotomy (DRT) and local treatment of the sciatic nerve with capsaicin, we found that SCT, DRT, and local capsaicin onto the sciatic nerve produced a significant inhibition of the BV-induced increase in volume of the injected paw, with a stronger effect of the SCT and the local capsaicin treatments than that of the DRT treatment. Static and dynamic mechanical allodynia in the BV test was assessed by measuring the paw withdrawal mechanical threshold and the paw withdrawal latency before and after the BV injection, respectively. Local capsaicin onto the sciatic nerve produced a significant inhibition of the BV-induced decrease in the paw withdrawal mechanical threshold, but not the paw withdrawal latency, of the injected paw. These findings suggest that neurogenic components, via dorsal root reflex and axon reflex mechanisms, are probably involved in the maintenance and the development of the BV-induced inflammation. In addition, the capsaicin-sensitive primary afferents may play differential roles in the development of the BV-induced static and dynamic mechanical allodynia.