Enzymes mediate diverse cancer processes by exerting their functional activities. Precise profiling of enzyme activity in the tumor microenvironment (TME) is therefore critical to understanding and targeting the pathological roles of enzymes in cancer. Here, we report biomodified nanoprobes for the highly sensitive detection of specific protease activities in medulloblastoma (MB) across scales. The surface-enhanced Raman scattering (SERS)-based nanoprobes use peptide-functionalized three-dimensional (3D) surfaces rich in electromagnetic hotspots to facilitate both enzymatic hydrolysis and plasmonic enhancement of Raman signals. We apply the nanoprobes to analyze multiple protease activities in 2D in vitro culture and 3D tumor cell spheroids, uncovering heterogeneous activity maps among different cell types, therefore allowing for recognition of distinct MB cellular subtypes. Through spatially resolved in situ localization of protease activity, we observe the suppressed protease function in the core region of tumor spheroids. Furthermore, in a pilot clinical assay (n = 27), the nanoprobe-based SERS assay reveals elevated levels of protease activity in sera of MB patients, and achieves great accuracy in discriminating MB from noncancer controls with the area under the receiver operating characteristic curve of 1.0. Together, this study offers a framework for functional, multiscale measurement of protease dysregulation in cancer.

Definitive diagnosis of intracranial tumors by less-invasive approaches provides diagnostic information to facilitate personalized treatment decisions and avoid unnecessary invasive biopsy operations. However, it remains challenging, largely due to a lack of reliable molecular biomarkers. Here, we demonstrated that molecular profiles of small extracellular vesicles (sEVs) in cerebrospinal fluid (CSF) decoded by surface-enhanced Raman spectroscopy (SERS) revealed highly specific signatures to detect and accurately discriminate common primary intracranial tumors in children that can be challenging to distinguish using standard-of-care imaging. Specifically, the fabrication of silver nanocube-based three-dimensional SERS substrates enabled the acquisition of highly resolved Raman spectra of sEVs in clinical CSF samples. The development of stacking machine learning frameworks to analyze the Raman data sets unveiled differential vibrational modes and generated high accuracy for diagnosing pediatric medulloblastoma (MB), the most common malignant brain tumor in children, with an area under the receiver operating characteristic curve (AUC(ROC)) of 0.963. Furthermore, we observed high discriminative capacity of our Raman spectral classifier to distinguish MB from other brain tumors (AUC(ROC) = 0.906). Finally, we showed that dynamic analysis of the CSF sEV Raman profiles allowed monitoring of the therapeutic response of MB at the molecular level. Our study holds promise for facilitating precision medicine in brain tumors.

MicroRNA (miRNA) dysregulation is closely related to the occurrence and progression of medulloblastoma (MB). However, the full potential of serum circulating miRNAs in MB diagnosis is restricted by their ultralow abundance in peripheral blood due to blood-brain barrier. Here, we report the direct preamplification-free detection of aberrant expression of oncogenic miRNAs in serum from MB patients by proposing a simple yet robust single-molecule assay that combines biphasic sandwich hybridization in nucleic acids and the dark-field single-particle plasmonic imaging (B2S2PI). In this strategy, signal DNA was prehybridized with target miRNA in homogeneous solution to form sDNA-RNA complexes. Then the captured DNA strands with rationally adjusted surface densities could efficiently capture the sDNA-RNA complexes to generate a well-separated DNA-RNA sandwich structure. The combination of homogeneous and heterogeneous reactions enabled interface-mediated hybridization reactions to maintain molecular stability with fewer bases, making it suitable for the direct amplification-free assays of short miRNA targets. Labeling the DNA-RNA hybrids with plasmatic gold nanotags allowed nondestructive recognition and imaging of individual miRNA targets under mild conditions with high signal-to-noise ratio. By digitally counting and analyzing the bright plasmonic resonant scattering spots, B2S2PI enabled both the measurement of a low femtomolar concentration of circulating miRNA-21 in 5 μL sample volume within a turnaround of 2 h and the discrimination of single base mismatches. Moreover, B2S2PI was universal for detecting miRNAs with different sequences and secondary structures. Further analysis of clinical serum samples revealed that B2S2PI was capable of accurately distinguishing MB patients from noncancer controls with an area under the curve (AUC) of 0.99, which was superior to that of qRT-PCR. B2S2PI holds promise as a novel alternative means for single-molecule miRNA assay and sheds light on the circulating nucleic acid-based liquid biopsy of intracranial malignant tumors.

Monitoring of cerebrospinal fluid (CSF) microRNAs (miRs) offers a promising option for the diagnosis and management of patients with central nervous system tumors. However, the sensitive detection of miRs in clinical CSF samples has been hindered by the ultra-low abundance of target miRs. Here, we report an electrochemical biosensor for the highly sensitive label-free detection of CSF miR-21 relying on target-induced redox signal amplification (eTIRSA). The biosensor was developed by covalently assembling the capture stands partially complementary to miR-21 on the gold nanoparticle-coated glassy carbon electrode. In the presence of miR-21, the short capture stand hybridized with the partial bases of miR-21, allowing the rest sequence of the target molecule to further bind with a long guanine-rich sequence which could specifically adsorb a number of methylene blue indicators, thus generating an amplified electrochemical redox signal, typically at a working potential of − 0.19 V (vs. SCE). The response of the surface-bound methylene blue indicators was positively correlated to the concentration of miR-21, providing a dynamic range of 0.5–80 pM and a limit of detection down to 56 fM. Moreover, the eTIRSA biosensor had high specificity with single-base resolution and exhibited good performance for label-free quantification of miR-21 in medulloblastoma cell extracts and clinical CSF samples and for accurate discrimination of medulloblastoma against non-cancer controls, indicating its potential application in CSF miR-based liquid biopsy of brain cancers.

The aim of this study was to investigate the role of BTBD10 in glioma tumorigenesis. The mRNA and protein levels of BTBD10 in 52 glioma tissues and eight normal brain tissues were determined using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively. U251 human glioblastoma cells were infected with BTBD10-expressing or control lentiviruses. Cell growth was evaluated using the methyl thiazolyl tetrazolium (MTT) assay. Cell apoptosis and cell cycle distribution were analyzed using flow cytometry. Cyclin D1 and p-Akt levels were determined using western blot analysis. The results showed that BTBD10 mRNA and protein levels were significantly lower in glioma tissues than in normal brain tissues. Additionally, BTBD10 levels were significantly lower in high-grade gliomas than in low-grade tumors. Compared with control cells, U251 cells overexpressing BTBD10 exhibited decreased cell proliferation, increased cell accumulation at the G0/G1 phase, increased cell apoptosis, and decreased levels of cyclin D1 and p-Akt. These findings show that BTBD10 is downregulated in human glioma tissue and that BTBD10 expression negatively correlates with the pathological grade of the tumor. Furthermore, BTBD10 overexpression inhibits proliferation, induces G0/G1 arrest, and promotes apoptosis in human glioblastoma cells by downregulating cyclin D1- and Akt-dependent signaling pathways.

The sensitive analysis of microRNAs (miRNAs) in cerebrospinal fluid (CSF) holds promise for the minimally invasive early diagnosis of brain cancers such as pediatric medulloblastoma but remains challenging due partially to a lack of facile yet sensitive sensing methods. Herein, an enzyme-free triple-signal amplification electrochemical assay for miRNA was developed by integrating the target-triggered cyclic strand-displacement reaction (TCSDR), hybridization chain reaction (HCR), and methylene blue (MB) intercalation. In this assay, the presence of target miRNA (miR-9) initiated the TCSDR and produced primers that triggered the subsequent HCR amplification to generate copious double-stranded DNAs (dsDNAs) on the electrode surface. Intercalation of a large number of MB reporters into the long nicked double helixes of dsDNAs yielded a more enhanced signal of differential pulse voltammetry. The enzyme-free multiple-amplification approach allowed for highly sensitive (detection limit: 6.5 fM) and sequence-specific (single-base mismatch resolution) detection of miR-9 from tumor cells and human CSF with minimal sample consumption (10 μL). Moreover, the clinical utilization of this method was documented by accurate discrimination of five medulloblastoma patients from the nontumoral controls. In light of its sensitivity, specificity, and convenience of use, this electrochemical method was expected to facilitate the early detection of malignant brain tumors.

Linear scleroderma is the most common type of localized scleroderma in children. Lesions rarely involve areas other than the skin, and nervous system involvement is even rare. We reported a case of a 6-year-old girl who was admitted to the hospital with recurrent seizures for 4 weeks. Before that, she had left frontal plaques for more than 1 year. Radiological imaging of the brain showed multiple abnormal lesions and skin biopsy of the plaques indicated scleroderma. After drug therapy, the girl had no recurrence of epilepsy, and no obvious abnormalities were found in the reexamination of neuroimaging. We performed further radiological examination on this patient and reviewed the literatures for this rare case.

Congenital teratomas are extremely rare and mainly midline tumors arising in the pineal regions in childhood brain tumors which are rarer cases occur in the lateral ventricle. Atrial septal defect (ASD) is detected in approximately 0.15% of newborns. We report an intracranial massive immature teratoma of the lateral ventricle in a 33-day-old infant on account of its rare location, comorbidity, and rapidly increasing size after surgery. Based on our information, this was the first case of congenital immature teratoma of the lateral ventricle comorbidity with ASD.

Intracranial bacterial infection remains a major cause of morbidity and mortality in neurosurgical cases. Metabolomic profiling of cerebrospinal fluid (CSF) holds great promise to gain insights into the pathogenesis of central neural system (CNS) bacterial infections. In this pilot study, we analyzed the metabolites in CSF of CNS infection patients and controls in a pseudo-targeted manner, aiming at elucidating the metabolic dysregulation in response to postoperative intracranial bacterial infection of pediatric cases. Untargeted analysis uncovered 597 metabolites, and screened out 206 differential metabolites in case of infection. Targeted verification and pathway analysis filtered out the glycolysis, amino acids metabolism and purine metabolism pathways as potential pathological pathways. These perturbed pathways are involved in the infection-induced oxidative stress and immune response. Characterization of the infection-induced metabolic changes can provide robust biomarkers of CNS bacterial infection for clinical diagnosis, novel pathways for pathological investigation, and new targets for treatment.

此前文献报道,有脑积水或蛛网膜囊肿病人行腹腔分流术后出现裂隙脑室综合征,表现为间歇性头疼以及脑室狭小.目前对裂隙脑室综合征的发病机理及治疗方法尚未深入,其对神经外科医生来说仍然是一个挑战,仍需进一步研究.综述了裂隙脑室综合征的病因、病理生理、临床表现、诊断及其治疗.

Background: To explore the specific prognosis related microRNAs (miRNAs) of glioma. Methods: The miRNA-Seq data and clinical information of glioma patients were downloaded from the TCGA (510 cases) and GEO (GSE112009, 25 cases) database. LASSO & COX regression was used to develop a miRNA-based model for predicting patient survival in the training set (n=255), to carry out glioma prognostic related miRNAs screening, and to construct a linear risk model based on the expression profiles of seven miRNAs. COX regression analysis was used to determine whether the miRNAs risk model was an independent prognostic factor. Results: Seven survival-related miRNAs (miR-140-5p, miR-145-5p, miR-148a-3p, miR-183-5p, miR222-3p, miR-223-3p, and miR-374a-5p) were identified in the training set. This showed that the overall survival time of the high-risk group was significantly lower than that of the low-risk group in the training set, prediction set, and validation set (P<0.05). Further analysis revealed that age and Karnofsky score both affected the risk of glioma. By crossing seven potential target genes of microRNAs, 620 effective target genes were obtained and GO analysis showed that these were related to the positive regulation of cell migration, neuron migration, and the response of transforming growth factor, and KEGG analysis showed they were related to the TGF-beta signaling pathway, MAPK signaling, and AGE-RAGE signaling pathway in diabetic complications. Conclusions: Seven miRNAs which regulate target genes to participate in related signaling pathways and lead to a poor prognosis were identified as biomarkers of glioma.

Cushing disease has a very high mortality rate and glucocorticoid resistance caused by GR down-regulation is one major reason of mortality. Although HIF1α signaling and GR signaling are involved in the pathogenesis of pituitary adenomas, it's unclear whether and how these two essential pathways could cross-talk with each other. Here, we performed a comprehensive study to investigate the reciprocal effects of HIF1α and GR on each other in AtT20 cell lines and explored the potential therapeutic effect of HIF1α inhibitor in in-vivo mouse model. We find that hypoxia up-regulated the promoter activity, mRNA and protein levels of GR and the induced GR protein was localized in cytosol. On the other hand, GR activation by its agonist DEX increased HIF1α protein through post-transcriptional mechanism. However, hypoxia and DEX show differential synergistic effects on HIF1α and GR. In hypoxia-DEX condition, HIF1α protein was further up-regulated but mainly localized in cytosol while GR was trapped and degraded in cytosol via UPS pathway. Further Co-IP experiments demonstrate that DNA binding domain of GR can interact with PASb domain of HIF1α. In a in-vivo mouse model of Cushing's disease, HIF1α inhibitor reduced HIF1α and GR protein levels, reduced tumor size and lowered the plasma concentrations of ACTH and corticosterone. In summary, we find that a novel HIF1α-GR crosstalk contributes to the pathogenesis of pituitary adenomas and HIF1α inhibitor shows potential therapeutic effects for Cushing's disease.

Reliable monitoring of metabolites in biofluids is critical for diagnosis, treatment, and long-term management of various diseases. Although widely used, existing enzymatic metabolite assays face challenges in clinical practice primarily due to the susceptibility of enzyme activity to external conditions and the low sensitivity of sensing strategies. Inspired by the micro/nanoscale confined catalytic environment in living cells, the coencapsulation of oxidoreductase and metal nanoparticles within the nanopores of macroporous silica foams to fabricate all-in-one bio-nanoreactors is reported herein for use in surface-enhanced Raman scattering (SERS)-based metabolic assays. The enhancement of catalytical activity and stability of enzyme against high temperatures, long-time storage or proteolytic agents are demonstrated. The nanoreactors recognize and catalyze oxidation of the metabolite, and provide ratiometric SERS response in the presence of the enzymatic by-product H2O2, enabling sensitive metabolite quantification in a "sample in and answer out" manner. The nanoreactor makes any oxidoreductase-responsible metabolite a candidate for quantitative SERS sensing, as shown for glucose and lactate. Glucose levels of patients with bacterial infection are accurately analyzed with only 20 µL of cerebrospinal fluids, indicating the potential application of the nanoreactor in vitro clinical testing.

目的 探讨儿童后颅窝肿瘤显微切除术中联合后组颅神经电生理监测(intraoperative posterior cranial nerves monitoring,IPCNM)对后组颅神经功能保护的影响. 方法 回顾性收集2012年3月至2015年12月由上海市儿童医院神经外科收治的经CT或MRI证实为后颅窝肿瘤患儿42例作为研究对象.将2012年3月至2013年8月收治的行单纯显微镜下肿瘤切除术的19例后颅窝肿瘤患儿纳入非IPCNM组,2013年9月至2015年12月收治的行显微镜下肿瘤切除过程中联合多模式神经电生理监测的23例后颅窝肿瘤患儿纳入IPCNM组.观察IPCNM组后颅窝肿瘤患儿手术中后组颅神经的实时情况,针对手术的进程给予适当辅助与及时指导;并随访所有后颅窝肿瘤患儿术后后组颅神经功能,分别比较两组间和各组组内后颅窝肿瘤患儿的术前、术后后组颅神经功能恢复情况;同时比较两组后颅窝肿瘤患儿在肿瘤全切率方面的差别. 结果 IPCNM组23例后颅窝肿瘤患儿,13例(56.52％)肿瘤全切;非IPCNM组19例后颅窝肿瘤患儿,10例(52.63％)肿瘤全切,差异无统计学意义(x2=0.098,P=0.752).同时在术后1周、4周、12周、24周、48周随访后组颅神经功能情况并进行组间和组内比较分析,显示IPCNM组在术后1周和4周与术前比较,差异具有统计学意义(P＜0.05);非IPCNM组在术后1周、4周和12周与术前比较,差异具有统计学意义(P＜0.05);IPCNM组和非IPCNM组在术后1周和4周的后组颅神经功能评估差异具有统计学意义(P＜0.05). 结论 在儿童后颅窝肿瘤手术中联合进行多模式的神经电生理监测不仅为手术时避免损伤后组颅神经提供了依据,而且可以有效保护后组颅神经功能,降低并发症发生率和手术风险,缩短术后后组颅神经功能的恢复时间.虽然术中联合电生理监测对于后颅窝肿瘤的切除提出了较为直观的作用,但并没有进一步提高肿瘤全切率.

Detection and inhibition of bacteria are universally required in clinics and daily life for health care. Developing a dual-functional material is challenging and in demand, engaging advanced applications for both defined bioanalysis and targeted biotoxicity. Herein, magnetic silver nanoshells are designed as a multifunctional platform for the detection and inhibition of bacteria. The optimized magnetic silver nanoshells enable direct laser desorption/ionization mass spectrometry based metabolic analysis of bacteria (≈10 µL-1 ), in complex biofluids. The serum infection process (0-10 h) is monitored by statistics toward clinical classification. Moreover, magnetic silver nanoshells facilitate surface adhesion on bacteria due to nanoscale surface roughness and thus display long-term antibacterial effects. Bacteria metabolism is studied with metabolic biomarkers (e.g., malate and lysine) identified during inhibition, showing cell membrane destruction and dysfunctional protein synthesis mechanisms. This work not only guides the design of material-based approaches for bioanalysis and biotoxicity, but contributes to bacteria-related diagnosis by using specific metabolic biomarkers for sensitive detection and new insights by monitoring metabolomic change of bacteria for antibacterial applications.

This work aimed at investigating the possibility and effectiveness of osteoinductive calcium phosphate (CaP) ceramics to close the drilled skull holes and prevent the postoperative cerebrospinal fluid (CSF) leaking in children's endoscopic neurosurgery. Five children patients (four boys and one girl, 3- to 8-years old) underwent the surgery, in which the endoscopic third ventriculostomy (ETV) was operated in four cases of hydrocephalus, and biopsy and ETV were both performed in one case of pineal tumor. The drilled skull holes were filled with the commercial osteoinductive CaP ceramics. The patients were followed up by CT scan at 1, 7 days, 3 and 6 months postoperatively. All the five cases were successful, and the holes were closed well after filled with the ceramics. The follow-up survey showed that no CSF leaking or rejection reaction was found. The CT scan indicated that the drilled holes began healing at 7 days postoperatively, and a relatively complete healing happened at 6 months postoperatively. The excellent ability of the CaP ceramics to induce bone regeneration was also confirmed by repairing the skull defects in a monkey model. The results of μ-CT and histological analysis showed that a bony structure with irregular array occurred at the defect area, and the newly formed bone volume density reached 65.7%. In conclusion, the osteoinductive CaP ceramics could be an ideal material to treat the drilled skull holes in children's endoscopic neurosurgery and prevent CSF leaking afterwards. However, further investigation with more cases and longer follow-up was required to evaluate the clinical effect.

Current metabolic analysis is far from ideal to engage clinics and needs rationally designed materials and device. Here we developed a novel plasmonic chip for clinical metabolic fingerprinting. We first constructed a series of chips with gold nanoshells on the surface through controlled particle synthesis, dip-coating, and gold sputtering for mass production. We integrated the optimized chip with microarrays for laboratory automation and micro-/nanoscaled experiments, which afforded direct high-performance metabolic fingerprinting by laser desorption/ionization mass spectrometry using 500 nL of various biofluids and exosomes. Further we for the first time demonstrated on-chip in vitro metabolic diagnosis of early stage lung cancer patients using serum and exosomes. This work initiates a new bionanotechnology based platform for advanced metabolic analysis toward large-scale diagnostic use.

Reliable profiling of the extracellular dopamine (DA) concentration in the central nervous system is essential for a deep understanding of its biological and pathological functions. However, quantitative determination of this neurotransmitter remains a challenge because of the extremely low concentration of DA in the cerebrospinal fluid (CSF) of patients. Herein, on the basis of the specific recognition of boronate toward diol and N-hydroxysuccinimide ester toward the amine group, a simple and highly sensitive strategy was presented for DA detection by using surface-enhanced Raman scattering (SERS) spectroscopy as a signal readout. This was realized by first immobilizing 3,3'-dithiodipropionic acid di( N-hydroxysuccinimide ester) on gold thin film surfaces to capture DA, followed by introducing 3-mercaptophenylboronic acid (3-MPBA)-functionalized silver nanoparticles to generate numerous plasmonic "hot spots" with the nanoparticle-on-mirror geometry. Such a dual-recognition mechanism not only avoids complicated bioelement-based manipulations but also efficiently decreases the background signal. With the direct use of the recognition probe 3-MPBA as a Raman reporter, the "signal-on" SERS method was employed to quantify the concentration of DA from 1 pM to 1 μM with a detection limit of 0.3 pM. Moreover, our dual-recognition-directed SERS assay exhibited a high resistance to cerebral interference and was successfully applied to monitoring of DA in CSF samples of patients.

In-vitro metabolite and drug detection rely on designed materials-based analytical platforms, which are universally used in biomedical research and clinical practice. However, metabolic analysis in bio-samples needs tedious sample preparation, due to the sample complexity and low molecular abundance. A further challenge is to construct diagnostic tools. Herein, we developed a platform using silver nanoshells. We synthesized SiO 2 @Ag with tunable shell structures by multi-cycled silver mirror reactions. Optimized nanoshells achieved direct laser desorption/ionization mass spectrometry in 0.5 μL of bio-fluids. We applied these nanoshells for disease diagnosis and therapeutic evaluation. We identified patients with postoperative brain infection through daily monitoring and glucose quantitation in cerebrospinal fluid. We measured drug distribution in blood and cerebrospinal fluid systems and validated the function of blood-brain/cerebrospinal fluid-barriers for pharmacokinetics. Our work sheds light on the design of materials for advanced metabolic analysis and precision diagnostics.

Objective To observe the clinical efficacy of early external ventricular drainage (EVD) for the treatment of premature and/or low-birth weight neonates with germinal matrix intraventricular hemorrhage (GM-IVH).Methods A total of 14 children with GM-IVH grade Ⅲ followed up for more than 24 months and admitted to the Department of Neurosurgery,Shanghai Children's Hospital,Shanghai Jiao Tong University from January 2013 to February 2014 were enrolled retrospectively.All children were treated with emergent lateral ventricle external drainage for continuous drainage of cerebrospinal fluid after the definite diagnosis.The closed cerebrospinal fluid (CSF) drainage system was applied to prevent secondary intracranial infection.Results The mean EVD time of 14 children was 11.9 ±1.7 d,the volume of CSF drainage was 33.0 ± 12.9 ml/d,and the intraventricular pressure (IVP) was controlled under 50 mmH2O.The intraventricular hemorrhage disappeared in all cases after EVD.The examination at 2 month after the removal of external ventricular drainage in 14 children,the ventricular system continued to expand in 8 cases.Then the ventriculo-peritoneal shunt (VPS) was performed.At 2 months after VPS,the ventricular system was reduced.After EVD,6 cases were followed up for 24 month,and the ventricular system was reduced.and none of them underwent VPS.Conclusion The early treatment of premature/low-birth weight neonates with GM-IVH may help to prevent the occurrence of hydrocephalus after bleeding and reduce the proportion of conducting VPS.