External validation of efficacy and reliability of different retrograde intrarenal surgery (RIRS) scoring systems for pediatric nephrolithiasis.

INTRODUCTION: Retrograde Intrarenal Surgery (RIRS) is one of minimally invasive procedures for pediatric upper urinary stones. However, a RIRS predictive system for children to evaluate postoperative stone free rate (SFR) is still unavailable. OBJECTIVE: The aim of this study is to validate the efficacy and reliability of different RIRS scoring systems for children. STUDY DESIGN: We collected clinical data of 137 pediatric patients treated with RIRS in our center between 2014 and 2021. All the predictors were acquired by preoperative non-contrast CT or CT urography. Receiver Operative Curve (ROC) and Area Under Curve (AUC) were showed to compare the predictive power of different models. RESULTS: A total of 162 RIRS procedures were performed for these 137 pediatric patients. Median surgical duration, irrigation volume and hospitalization were 30 (20, 40) min, 500 (300, 1000) ml and 6 (4, 7) days, respectively. Overall SFR and complication rate was 79.6% (129/162) and 29.2% (40/137), respectively. Significant difference was detected between non-stone free group and stone free group in terms of stone complexity (p < 0.001), cumulative stone sizes [2.3 (2.0, 3.5) cm vs. 1.5 (1.0, 2.0) cm, p < 0.001], RUS groups (p < 0.001), S-ReSC groups (p < 0.001) and RIRS nomogram score [10 (8, 12) vs. 23 (18, 25), p < 0.001]. Among them, RIRS nomogram presented with the maximum AUC values in comparison with the other two systems (RUS: 0.944 vs. 0.874, p = 0.001; S-ReSC: 0.944 vs. 0.808, p < 0.001). DISCUSSION: We reported the largest sample size of pediatric patients treated with RIRS in our center. Similar with previous studies, RIRS is an efficacious and safe option for pediatric patients. RIRS nomogram showed the best predictive outcome due to the inclusion of multiple parameters, but an innovative predictive system based on pediatric clinical data is warranted in the future. CONCLUSION: Among the three scoring systems, RIRS nomogram showed the most optimal predictive power of postoperative SFR for pediatric patients.

LPS alters the immuno-phenotype of glioma and glioma stem-like cells and induces in vivo antitumor immunity via TLR4.

BACKGROUND: This study examined the ability of lipopolysaccharide (LPS) to affect glioma and glioma stem-like cells (GSCs) in vitro and to induce antitumor immunity in vivo and the role of TLR4 in these processes. METHODS: Using RT-PCR and immunohistochemistry, we examined the expression of TLR4 in 34 glioblastoma clinical samples. Using real time-PCR, western blot and ELISA analyses, the effect of LPS stimulation on the expression of immune related molecules was evaluated in RG2 and U87 GSCs. Control or LPS-pretreated RG2 GSCs were intracranially or subcutaneously implanted into wild-type or nude Fisher 344 rats. Histopathological examinations were used to assess tumor progression and immune infiltration and Kaplan-Meier analyses to compare survival times of the animal models. RESULTS: TLR4 was highly expressed in glioblastoma clinical samples. In vitro LPS stimulation for 6 h significantly altered expression of immune related molecules in RG2 and U87 GSCs. However, prolonged LPS stimulation diminished this effect. Rats inoculated intracranially with LPS-pretreated RG2 GSCs survived significantly longer than rats inoculated with control RG2 GSCs. In vivo, LPS-pretreated RG2 GSCs expressed higher levels of MHC molecules, CXCL10 and TNF-α and recruited more CD8+ lymphocytes. However, intratumoral LPS treatment was not equally beneficial. Furthermore, the in vitro and in vivo effects of LPS stimulation appeared to be largely TLR4-dependent. CONCLUSION: LPS pretreatment promotes the recognition and eradication of tumor GSCs in vivo when the immune function of the tumor-bearing host is intact. In addition, our data indicate a complex relationship between bacterial infection and glioma prognosis.

Lithium enhances the antitumour effect of temozolomide against TP53 wild-type glioblastoma cells via NFAT1/FasL signalling.

BACKGROUND: We previously showed that activation of the nuclear factor of activated T cells (NFAT)1/Fas ligand (FasL) pathway induces glioma cell death. Lithium (Li) is an inhibitor of glycogen synthase kinase (GSK)-3 that activates NFAT1/FasL signalling. Temozolomide (TMZ) inhibits GSK-3 and activates Fas in tumour protein (TP)53 wild-type (TP53wt) glioma cells. The present study investigated the combinational effects of TMZ and low-dose Li on TP53wt glioma cells. METHODS: The combined effect of TMZ and Li was examined in TP53wt U87 and primary glioma cells and a mouse xenograft model. RESULTS: Combination with 1.2 mM Li potentiated TMZ-induced cell death in TP53wt glioma cells, as determined by neurosphere formation and apoptosis assays. Temozolomide combined with Li treatment inhibited GSK-3 activation, promoted NFAT1 nuclear translocation and upregulated Fas/FasL expression. Targeted knockdown of NFAT1 expression blocked the induction of cell death by TMZ and Li via FasL inhibition. In vivo, combined treatment with TMZ and Li suppressed tumour growth and prolonged the survival of tumour-bearing mice. However, the combination of TMZ and Li did not produce a statistically significant effect in TP53mut glioma cells. CONCLUSIONS: Temozolomide combined with low-dose Li induces TP53wt glioma cell death via NFAT1/FasL signalling. This represents a potential therapeutic strategy for TP53wt glioma treatment.

Monomeric annexin A2 is an oxygen-regulated toll-like receptor 2 ligand and adjuvant.

BACKGROUND: Annexin A2 (ANXA2) is a pleiotropic, calcium-dependent, phospholipid-binding protein with a broad tissue distribution. It can be intracellular, membrane-bound, or secreted, and it exists as a monomer or heterotetramer. The secreted ANXA2 heterotetramer signals human and murine macrophages to produce IL-1, IL-6, and TNF-α through TLR4/MyD88- and TRIF-dependent pathways. METHODS: GL261 glioma cells were cultured in 5 % or 20 % O2. Monomeric ANXA2 (ANXA2m) was identified as a TLR2-binding protein enriched in 5 % O2 by mass spectrometry. Purified ANXA2m and ANXA2-derived peptides were added to TLR2-expressing reporter cells and immature dendritic cells (DCs) cells in vitro. ANXA2m was then mixed with chicken ovalbumin (OVA) for vaccination of TLR2 (+/+) and TLR2 (-/-) mice for subsequent quantification of antigen-specific CD8(+) T cell responses. The TLR2-binding region of ANXA2m was determined using various peptides derived from the ANXA2 amino terminus on TLR2 reporter cells and in vaccinated mice. RESULTS: ANXA2m is overexpressed by murine glioblastoma GL261 cells grown under 5 % O2, but not atmospheric 20 % O2, and acts as an adjuvant by inducing murine and human DC maturation through TLR2. ANXA2m upregulates CD80 and CD86 expression, enhances antigen cross-presentation, and induces the secretion of IL-12p70, TNF-α, and IFN-γ. The amino-terminal 15 amino acids of ANXA2m are necessary and sufficient for TLR2 binding and DC activation. CONCLUSION: This novel finding adds to the known functions of ANXA2 and suggests ways to exploit it as a vaccine adjuvant. ANXA2-antigen fusion peptides could be developed for patients as "off-the-shelf" agents containing common tumor antigens. Alternatively, they could be "personalized" and synthesized after tumor sequencing to identify immunogenic tumor-specific neo-antigens. As the amino terminal 15 amino acids of ANXA2 are required to stimulate TLR2 activity, a fusion peptide could be as short as 30 amino acids if one or two CD8 T cell epitopes are fused to the ANXA2 amino terminal portion. Future work will address the efficacy of ANXA2 peptide fusions alone and in combination with established TLR agonists to induce synergy in preclinical models of glioma as observed in other vaccines.

CD200 in CNS tumor-induced immunosuppression: the role for CD200 pathway blockade in targeted immunotherapy.

BACKGROUND: Immunological quiescence in the central nervous system (CNS) is a potential barrier to immune mediated anti-tumor response. One suppressive mechanism results from the interaction of parenchyma-derived CD200 and its receptor on myeloid cells. We suggest that CD200/CD200R interactions on myeloid cells expand the myeloid-derived suppressor cell (MDSC) population and that blocking tumor-derived CD200 will enhance the efficacy of immunotherapy. METHODS: CD200 mRNA expression levels in human brain tumor tissue samples were measured by microarray. The amount of circulating CD200 protein in the sera of patients with brain tumors was determined by ELISA and, when corresponding peripheral blood samples were available, was correlated quantitatively with MDSCs. CD200-derived peptides were used as competitive inhibitors in a mouse model of glioblastoma immunotherapy. RESULTS: CD200 mRNA levels were measured in human brain tumors, with different expression levels being noted among the sub groups of glioblastoma, medulloblastoma and ependymoma. Serum CD200 concentrations were highest in patients with glioblastoma and correlated significantly with MDSC expansion. Similarly, in vitro studies determined that GL261 cells significantly expanded a MDSC population. Interestingly, a CD200R antagonist inhibited the expansion of murine MDSCs in vitro and in vivo. Moreover, inclusion of CD200R antagonist peptide in glioma tumor lysate-derived vaccines slowed tumor growth and significantly enhanced survival. CONCLUSION: These data suggest that CNS-derived tumors can evade immune surveillance by engaging CD200. Because of the homology between mouse and human CD200, our data also suggest that blockade of CD200 binding to its receptor will enhance the efficacy of immune mediated anti-tumor strategies for brain tumors.

Vaccine injection site matters: qualitative and quantitative defects in CD8 T cells primed as a function of proximity to the tumor in a murine glioma model.

Malignant gliomas are lethal brain tumors for which novel therapies are urgently needed. In animal models, vaccination with tumor-associated Ags efficiently primes T cells to clear gliomas. In clinical trials, cancer vaccines have been less effective at priming T cells and extending survival. Generalized immune suppression in the tumor draining lymph nodes has been documented in multiple cancers. However, a systematic analysis of how vaccination at various distances from the tumor (closest to farthest) has not been reported. We investigated how the injection site chosen for vaccination dictates CD8 T cell priming and survival in an OVA-transfected murine glioma model. Glioma-bearing mice were vaccinated with Poly:ICLC plus OVA protein in the neck, hind leg, or foreleg for drainage into the cervical, inguinal, or axillary lymph nodes, respectively. OVA-specific CD8 T cell number, TCR affinity, effector function, and infiltration into the brain decreased as the vaccination site approached the tumor. These effects were dependent on the presence of the tumor, because injection site did not appreciably affect CD8 T cell priming in tumor-free mice. Our data suggest the site of vaccination can greatly impact the effectiveness of cancer vaccines. Considering that previous and ongoing clinical trials have used a variety of injection sites, vaccination site is potentially a critical aspect of study design that is being overlooked.

Vasculogenic mimicry-potential target for glioblastoma therapy: an in vitro and in vivo study.

Glioblastoma is one of the most angiogenic human tumors and characterized by microvascular proliferations. A better understanding of glioblastoma vasculature is needed to optimize anti-angiogenic therapy that has shown a promising but incomplete efficacy. The present study examined 48 glioblastomas by CD34 endothelial marker periodic acid-Schiff (PAS) dual staining and found non-endothelial cell-lined blood vessels that were formed by tumor cells (vasculogenic mimicry, VM) existing in a fraction of these tumors. We hypothesized that CD133-positive glioblastoma stem-like cells (GSCs) may play a pivotal role in glioblastoma VM formation and then demonstrated in vitro and in vivo that a subset of GSCs were capable of vasculogenesis. Moreover, we found that several growth factors involved in normal angiogenesis were expressed in GSCs. We describe here a new mechanism of alternative glioblastoma vascularization and open a new perspective for the anti-vascular treatment strategy.

Burr-hole craniotomy treating chronic subdural hematoma: a report of 398 cases.

OBJECTIVE: To investigate the treatment of chronic subdural hematoma (CSDH) with burr-hole craniotomy in our hospital. METHODS: From January 2004 to December 2009, 398 patients with CSDH, 338 males and 60 females (male/female equal to 5.63/1), received burr-hole craniotomy in our hospital. The median age was 60 years with the mean age of (58.1 ± 18.1) years, (65.0 ± 14.5) years for females and (57.0 ± 18.2) years for males. Trauma history was determined in 275 patients (69.1%). Burr-hole craniotomy was performed under local anesthesia in 368 patients and general anesthesia in 30 patients. CSDH was classified into 3 groups according to the density on CT scan. Clinical data concerning etiologies, symptoms and signs, concomitant diseases, diagnosis, therapies and outcomes were investigated retrospectively. Patients'neurological status on admission and at discharge was also classified to judge the outcomes. RESULTS: Generally, trauma history showed few differences between those over 60 years old and under 60 years old, but showed obvious differences when gender was taken into account. Totally 123 male patients (60.0% of 204 cases) suffering from head injuries were under 60 years, whereas 35 female patients (85.4% of 41 cases) with trauma histories were over 60 years. The duration from trauma to appearance of clinical symptoms was (84.0 ± 61.7) days (range, 0-1493 days). Traumatic accident was the leading etiology, other accompanying diseases such as cerebral vascular disease, hypertension, etc, were also predisposing factors. Commonly, the elderly presented with hemiplegia/hemidysesthesia/hemiataxia and the young with headache. Most CSDH patients (95.6%) treated with burr-hole craniotomy successfully recovered. However, postoperative complications occurred in 17 cases, including recurrence of CSDH in 15 cases, subdural abscess in 1 case and pneumonia in 3 cases. CONCLUSION: Burr-hole craniotomy is an easy, efficient and reliable way to treat CSDH.