Direct Expression of Fluorinated Proteins in Human Cells for 19F In-Cell NMR Spectroscopy.

In-cell NMR spectroscopy is a powerful approach to study protein structure and function in the native cellular environment. It provides precious insights into the folding, maturation, interactions, and ligand binding of important pharmacological targets directly in human cells. However, its widespread application is hampered by the fact that soluble globular proteins often interact with large cellular components, causing severe line broadening in conventional heteronuclear NMR experiments. 19F NMR can overcome this issue, as fluorine atoms incorporated in proteins can be detected by simple background-free 1D NMR spectra. Here, we show that fluorinated amino acids can be easily incorporated in proteins expressed in human cells by employing a medium switch strategy. This straightforward approach allows the incorporation of different fluorinated amino acids in the protein of interest, reaching fluorination efficiencies up to 60%, as confirmed by mass spectrometry and X-ray crystallography. The versatility of the approach is shown by performing 19F in-cell NMR on several proteins, including those that would otherwise be invisible by 1H-15N in-cell NMR. We apply the approach to observe the interaction between an intracellular target, carbonic anhydrase 2, and its inhibitors, and to investigate how the formation of a complex between superoxide dismutase 1 and its chaperone CCS modulates the interaction of the chaperone subunit with the cellular environment.

Infiltrative and drug-resistant slow-cycling cells support metabolic heterogeneity in glioblastoma.

Metabolic reprogramming has been described in rapidly growing tumors, which are thought to mostly contain fast-cycling cells (FCCs) that have impaired mitochondrial function and rely on aerobic glycolysis. Here, we characterize the metabolic landscape of glioblastoma (GBM) and explore metabolic specificities as targetable vulnerabilities. Our studies highlight the metabolic heterogeneity in GBM, in which FCCs harness aerobic glycolysis, and slow-cycling cells (SCCs) preferentially utilize mitochondrial oxidative phosphorylation for their functions. SCCs display enhanced invasion and chemoresistance, suggesting their important role in tumor recurrence. SCCs also demonstrate increased lipid contents that are specifically metabolized under glucose-deprived conditions. Fatty acid transport in SCCs is targetable by pharmacological inhibition or genomic deletion of FABP7, both of which sensitize SCCs to metabolic stress. Furthermore, FABP7 inhibition, whether alone or in combination with glycolysis inhibition, leads to overall increased survival. Our studies reveal the existence of GBM cell subpopulations with distinct metabolic requirements and suggest that FABP7 is central to lipid metabolism in SCCs and that targeting FABP7-related metabolic pathways is a viable therapeutic strategy.

[Relationship between dynamic changes of psychological distress and quality of life in Chinese early breast cancer patients].

Objective: To explore the dynamic changes of Distress Thermometer scores and the relationship between psychological distress and quality of life in Chinese early breast cancer patients during chemotherapy. Methods: This prospective study enrolled 110 Chinese postoperative early breast cancer patients between March 2019 and December 2019. The psychological distress and quality of life (QOL) of patients were assessed by using the psychological distress management screening tool and the patient quality of life scale. Logistic regression model was used to analyze the influencing factors of psychological distress degree. The correlation between distress thermometer (DT) score changes and quality of life was analyzed by Pearson correlation analysis. Results: In total, 96 valid cases were analyzed. Before chemotherapy, 47 cases (49.0%) had DT score ≥4 points. After 2 cycles of chemotherapy, 40 cases (41.7%) had DT score ≥4 points. Thirty-four patients (35.4%) had DT score ≥4 points after chemotherapy. The DT score after chemotherapy was lower than that before chemotherapy and after 2 cycles of chemotherapy. Univariate analysis showed that income level and pathological stage were still significant related to the detection of DT score ≥4 points after chemotherapy (P<0.05). The changes of DT scores before and after chemotherapy were negatively correlated with the changes of quality of life ( r=-0.298, P=0.003). Conclusions: The detection rate of psychological distress in patients with early breast cancer during chemotherapy showed a decreasing trend. Income level and tumor stage are significant factors affecting the psychological distress of patients. There is a significant correlation between the psychological distress and the quality of life during chemotherapy. We should pay attention to the evaluation and monitoring state of psychological distress of patients during chemotherapy.

Unfolding spatiotemporal dynamics through symmetry reduction based on orbit topology.

Understanding key patterns in a spatially extended system is an essential task of modern physics of complex systems. Just like in low-dimensional nonlinear systems, here we show that orbit topology plays a critical role even for the investigation of spatiotemporal dynamics. First, we design a new scheme to reduce possible continuous symmetries that are prevailing in these systems based on topological consideration. The scheme is successfully demonstrated in the well-known pattern formation systems. Interesting bifurcation routes to chaos are conveniently revealed after symmetry reduction. In particular, we find that near the onset of turbulent dynamics, with an increase of instability, local phase chaos with the same spatial topological index may merge into more complex ones, while those with different indices induce defect chaos necessarily through connections docked with defects. The topological argument is so strong that the scenario presented here should be omnipresent in diverse systems.

Biological Characteristics and Genetic Diversity of Phomopsis asparagi, Causal Agent of Asparagus Stem Blight.

Asparagus stem blight is a regional disease. In the present study, we compared strains of Phomopsis asparagi from six different provinces to determine their biological characteristics and genetic diversity, differences in the pycnidium and conidium production, pathogenicity, and growth rate. Considerable differences were established in the pycnidium and conidium production among the P. asparagi strains from the six studied provinces. The largest pycnidium and conidium production had the strains from Fujian, followed by those from Hainan. The virulence of P. asparagi strains was significantly different but without a correlation with the geographical source of the strain. FJ2 had the highest virulence, followed by HN2, SD4, and SD5, whereas SD5 had the lowest virulence. The colony diameter and dry weight of the strains of asparagus stem blight fungus from the six provinces were substantially different. The colonies of HN1-5 had the largest diameters, whereas those of XT1-5, LT1-3, FJ1-5, and SX6 had smaller diameters. Four primers with good repeatability and strong specificity were selected from 100 intersimple sequence repeat (ISSR) primers. ISSR-PCR amplification was performed on 36 strains of asparagus stem blight fungus, and a large number of repeatable DNA fingerprints were obtained. Most of the amplified fragments were within 300 to 500 bp. In all, 69 total points, 64 multiple points, and 92.75% polymorphism points were established. The number of ISSR gene sites detected by four primers ranged from 14 to 20, with an average of 16 multiple sites. The copolymerization was divided into three groups: XT1-5, LT1-3, and FJ1-5, which were clustered into the first group; SD1-6, SX1-6, and HB1-6, clustered into the second group; and HN1-5 in the third group. The results of the cluster analysis revealed that the strains of the neighboring provinces had a nearer phylogenetic relationship than that between distant ones. Therefore, the system evolution of P. asparagi is related to the geographical distribution of its strains.

[The effect of ovarian function suppression on the psychological status of breast cancer patients under 35 years old in China].

Objective: To explore whether the addition of ovarian function suppression in endocrine therapy of Chinese breast cancer patients under 35 years old will affect the psychological state. Methods: This cross-sectional study enrolled 91 Chinese postoperative breast cancer patients aged 35 years or younger. The depression and anxiety state of patients were assessed by Patient Health Questionnaire-9 (PHQ-9) and General Anxiety Disorder-7 (GAD-7), and their sociodemographic characteristics were collected. Results: Among the 91 patients, 61 were receiving ovarian function suppression (OFS) treatment, 30 were not. Among the 30 patients with out OFS treatment, 2 had PHQ-9 score ≥8, 28 had PHQ-9 score < 8, 1 had GAD-7 score ≥10, and 29 had GAD-7 score < 10. Among the 61 patients with OFS, 19 had PHQ-9 score ≥8, 42 had PHQ-9 score < 8, 8 had GAD-7 score ≥10, and 53 had GAD-7 score <10. The incidence of depression was 6.7% and 31.1% in the non-OFS group and OFS group, respectively (P=0.018). The incidence of anxiety in the two groups was 3.3% and 13.1%, respectively (P=0.174). Univariate analysis showed that the incidence of depression was significantly higher in patients with OFS (P=0.018). After taking into account the sociodemographic factors, pathological stage and treatment of the patients, multivariate analysis showed that the administration of OFS was still significantly related to the incidence of depression (OR=9.14, 95% CI=1.52~55.16, P=0.016). There was no significant difference in the incidence of anxiety (P=0.174). Conclusions: For Chinese young breast cancer patients under 35 years old, the use of OFS in the adjuvant endocrine therapy may lead to a significant increase in the incidence of depression. We should pay attention to the evaluation and monitoring of the psychological state of this population.

[The clinical reports on adrenal insufficiency of patients with advanced solid tumors accepting anti-PD-1 antibody, SHR-1210 therapy].

Objective: To investigate the adrenocortical function changes of patients with advanced solid tumors who received the anti- programmed cell death protein-1 (PD-1) antibody, SHR-1210 therapy. Methods: The clinical data of 98 patients with advanced solid tumors who were enrolled in a prospective phase I trial of SHR-1210 therapy at our institution between April 27, 2016 and June 8, 2017 were collected. The levels of plasma adrenocorticotropic hormone (ACTH) and cortisol were evaluated in 96 patients. The clinical manifestations, laboratory tests and radiologic data were collected to define the immune-related adrenal insufficiency. Results: Until December 14th, 2018, no SHR-1210 related primary adrenal insufficiency occurred, and the incidence of immune-related secondary adrenal insufficiency was 1.0% among the 96 patients, which was identified as grade 2. No patient developed grade 3-4 adrenal insufficiency. The main clinical manifestations of the patient who was diagnosed as secondary adrenal insufficiency were grade 2 fatigue, anorexia and headache.The patient developed fatigue and anorexia at the 267th day after receiving the first dose of SHR-1210, the hypocortisolism occurred on the 279th day, and the headache emerged on the 291th day. The anorexia of patient who treated by physiological replacement doses of glucocorticoid since the 457th day was attenuated.The patient whose cortisol level was still below the normal limit continued to accept the hormone replacement therapy up to 776 days after the initial administration of SHR-1210. Conclusions: The incidence of SHR-1210 related adrenal insufficiency of patients with advanced solid tumors is low, and the symptoms can be effectively ameliorated by hormone replacement therapy. The potential adverse outcome of adrenal insufficiency following immunotherapy should be noticed by clinicians to avoid the occurrence of adrenal crisis.

Immunotherapy for Brain Tumors.

OPINION STATEMENT: Despite aggressive surgery, radiation, and systemic chemotherapy, the prognosis for patients diagnosed with malignant brain tumors remains extremely poor, and standard treatments carry significant risks for long-term neurocognitive deficits. There is a clear and urgent need for the development of more effective treatments that will add minimal toxicity to standard therapies for invasive brain cancers. Cancer immunotherapy is a treatment modality that holds promise for the delivery of tumor-specific cytotoxicity, with the potential to eliminate brain tumor cells without harming the eloquent brain.

[Clinical observation of thyroid-related adverse events induced by anti-PD-1 antibody SHR-1210 in patients with advanced solid tumor].

Objective: To assess the incidence and characteristics of thyroid dysfunction during anti-Programmed cell death 1 receptor (PD-1) antibody SHR-1210 therapy in patients with advanced solid tumor. Methods: The medical records of 98 patients who initiated SHR-1210 treatment between April 27, 2016 and June 8, 2017 in the phase 1 trial to evaluate the safety, efficacy, and pharmacokinetics of SHR-1210 in patients with advanced solid tumors were retrospectively reviewed. Serological tests of thyroid stimulating hormone (TSH) and free thyroxine (fT4) were measured at baseline and prior to each SHR-1210 administration. Results: A total of 86 patients had normal thyroid function before the first dose of SHR-1210 treatment. Nine out of 86 (10.5%) patients developed new onset hypothyroidism from euthyroid state. 12 patients presented thyroid dysfunction at baseline, 10 of whom were subclinical hypothyroid and 2 were hypothyroidism. Four out of 10 patients developed hypothyroidism from subclinical hypothyroid. Most patients with hypothyroidism were asymptomatic. Thyroid dysfunction occurred early (median, 55days) after the initiation of SHR-1210. The severity of hypothyroidism were all grade 1-2. No grade 3-4 hypothyroidism occurred. No patients discontinue the treatment of SHR-1210 due to clinical impact of the thyroid dysfunctions. Conclusions: Thyroid-related adverse events were common during anti-PD-1 antibody SHR-1210 treatment . The incidence of hypothyroidism is lower in patients with euthyroid state than in patients with thyroid dysfunction at baseline during SHR-1210 treatment . Thyroid function can be improved after thyroid hormone replacement. During SHR-1210 treatment, it is necessary to pay attention to monitor the thyroid function, especially in the patients with thyroid dysfunction at baseline. Trial registration: Chinese Clinical Trial Registry, 2016L01455.

Community fluctuations and local extinction in a planktonic food web.

Determining statistical patterns irrespective of interacting agents (i.e. macroecology) is useful to explore the mechanisms driving population fluctuations and extinctions in natural food webs. Here, we tested four predictions of a neutral model on the distribution of community fluctuations (CF) and the distributions of persistence times (APT). Novel predictions for the food web were generated by combining (1) body size-density scaling, (2) Taylor's law and (3) low efficiency of trophic transference. Predictions were evaluated on an exceptional data set of plankton with 15 years of weekly samples encompassing c. 250 planktonic species from three trophic levels, sampled in the western English Channel. Highly symmetric non-Gaussian distributions of CF support zero-sum dynamics. Variability in CF decreased while a change from an exponential to a power law distribution of APT from basal to upper trophic positions was detected. Results suggest a predictable but profound effect of trophic position on fluctuations and extinction in natural communities.

FOXO1 mediates RANKL-induced osteoclast formation and activity.

We have previously shown that the transcription factor FOXO1 is elevated in conditions with high levels of bone resorption. To investigate the role of FOXO1 in the formation of osteoclasts, we examined mice with lineage-specific deletion of FOXO1 in osteoclast precursors and by knockdown of FOXO1 with small interfering RNA. The receptor activator for NF-κB ligand (RANKL), a principal bone-resorbing factor, induced FOXO1 expression and nuclear localization 2 d after stimulation in bone marrow macrophages and RAW264.7 osteoclast precursors. RANKL-induced osteoclast formation and osteoclast activity was reduced in half in vivo and in vitro with lineage-specific FOXO1 deletion (LyzM.Cre(+)FOXO1(L/L)) compared with matched controls (LyzM.Cre(-)FOXO1(L/L)). Similar results were obtained by knockdown of FOXO1 in RAW264.7 cells. Moreover, FOXO1-mediated osteoclast formation was linked to regulation of NFATc1 nuclear localization and expression as well as a number of downstream factors, including dendritic cell-specific transmembrane protein, ATP6vod2, cathepsin K, and integrin αv. Lastly, FOXO1 deletion reduced M-CSF-induced RANK expression and migration of osteoclast precursors. In the present study, we provide evidence that FOXO1 plays a direct role in osteoclast formation by mediating the effect of RANKL on NFATc1 and several downstream effectors. This is likely to be significant because FOXO1 and RANKL are elevated in osteolytic conditions.

[The treatment and prognosis of peripheral primitive neuroectodermal tumor].

Objective: To evaluate the treatment and prognosis of peripheral primitive neuroectodermal tumor (pPNET). Methods: From March 2006 to April 2015, 47 patients with pPNET who had undergone chemotherapy in our hospital were enrolled. The clinical data and survival information of these patients were collected and interpreted retrospectively to analyze the effect of each treatment on the survival of patients. Results: The median overall survival (OS) for whole group was 23.5 months, and 5-year survival rate was 33.8%. In the patients who underwent radical surgery, the median OS was 70.4 months, the 5-year survival rate was 54.4%, the median disease-free survival (DFS) was 23.1months, and 5-year DFS rate was 34.4%. Sixteen patients had recurrences or metastasis after surgery. Eighty-one percent of them (13/16) occurred within 2 years after surgery. The difference of median OS between patients who got adjuvant chemotherapy and those who did not was statistically significant (P=0.04). But the difference of median PFS between these two groups was not statistically significant (P=0.057). There was no statistically significant difference for median OS (P=0.619) and median DFS (P=0.191) between patients who got adjuvant radiotherapy and those who did not. The recurrence rate between these two groups was not statistically significant (P=0.40). The median OS and PFS for 34 patients who received first-line palliative chemotherapy was 10.7 months and 3.2 months. 1-year and 2-year survival rates were 48.0% and 17.8%. The response rate and clinical benefit rate for first-line chemotherapy was 53.1% and 75.0%. The median PFS and OS for patients who received platinum-based regimens were 3.3 months and 14.5 months. The median PFS and OS for patients who got non-platinum regimens were 2.7 months and 10.3 months. There was no significant difference of PFS and OS between platinum-based and non-platinum regimens. Palliative surgery and radiotherapy did not improve the OS of pPNET this cohort. Conclusions: Comprehensive treatment including chemotherapy, radiotherapy and surgery is the standard treatment model for early pPNET patients. Adjuvant chemotherapy significantly improved the overall survival of early pPNET patients. Chemotherapy is the main treatment for patients with advanced pPNET. Platinum-based chemotherapy seem to be a good option.

Defining a new role of GW182 in maintaining miRNA stability.

GW182 binds to Argonaute (AGO) proteins and has a central role in miRNA-mediated gene silencing. Using lentiviral shRNA-induced GW182 knockdown in HEK293 cells, this study identifies a new role of GW182 in regulating miRNA stability. Stably knocking down GW182 or its paralogue TNRC6B reduces transfected miRNA-mimic half-lives. Replenishment of GW182 family proteins, as well as one of its domain Δ12, significantly restores the stability of transfected miRNA-mimic. GW182 knockdown reduces miRNA secretion via secretory exosomes. Targeted siRNA screening identifies a 3'-5' exoribonuclease complex responsible for the miRNA degradation only when GW182 is knocked down. Immunoprecipitation further confirms that the presence of GW182 in the RISC complex is critical in protecting Argonaute-bound miRNA.

CT and MRI features of patients with diastematomyelia.

OBJECTIVE: To evaluate computed tomography (CT) and magnetic resonance imaging (MRI) features in patients with diastematomyelia and to investigate clinical characteristics of this lesion. STUDY DESIGN: A retrospectively study. SETTING: The Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University. METHODS: A total of 82 diastematomyelia cases were retrospectively studied. All the patients underwent neurological examinations as well as MRI and CT of the spine. A self-established neurological functional grading system was used, and posterior tibial nerve somatosensory cortical-evoked potential (PTNSCEP) was measured to assess the neurological status of the patients. Imaging features of symmetry of splitting, presence of septum, location of lesion and number of split segments were studied. The neurological functional grading, PTNSCEP, and imaging findings were then analyzed and compared, and the difference was considered to be significant if P-value was lower than 0.05. RESULTS: Neurological functional grading and latency of PTNSCEP were significantly different but related in terms of symmetry of splitting, presence of septum and location of lesion. Although no significant differences were present in the number of split segments, the severity of the neurological functional grading and PTNSCEP impairment were not related to the number of split segments. CONCLUSION: The imaging features in diastematomyelia are characteristic and relate well with the clinical manifestations according to neurological functional grading and PTNSCEP measurement, except the number of split segments.

Controlling the incorporation of fluorinated amino acids in human cells and its structural impact.

Fluorinated aromatic amino acids (FAAs) are promising tools when studying protein structure and dynamics by NMR spectroscopy. The incorporation FAAs in mammalian expression systems has been introduced only recently. Here, we investigate the effects of FAAs incorporation in proteins expressed in human cells, focusing on the probability of incorporation and its consequences on the 19 F NMR spectra. By combining 19 F NMR, direct MS and x-ray crystallography, we demonstrate that the probability of FAA incorporation is only a function of the FAA concentration in the expression medium and is a pure stochastic phenomenon. In contrast with the MS data, the x-ray structures of carbonic anhydrase II reveal that while the 3D structure is not affected, certain positions lack fluorine, suggesting that crystallization selectively excludes protein molecules featuring subtle conformational modifications. This study offers a predictive model of the FAA incorporation efficiency and provides a framework for controlling protein fluorination in mammalian expression systems.

Invasive and prognostic significance of pRB in esophageal squamous cell carcinoma: a meta-analysis.

This paper investigates the association between protein retinoblastoma (pRB) loss and the T,N stage and prognosis in esophageal squamous cell carcinomas (ESCCs) using meta-analysis. We conducted a meta-analysis of 16 studies, comprising 1,117 patients to clarify this issue. All the studies searched by the electronic literature PubMed and http://www.KJEBM.com, which had been published during the period from January 1996 to January 2012 according to the inclusion criteria. Summary odds ratios (OR) were calculated using fixed or random-effects models. The summary odds ratios (ORs) for pRB inactive were 0.64 (95% confidence interval [CI]:0.45-0.91, P = 0.01) for T1/T2 versus T3/T4 tumors; summary OR = 0.69 (95% CI:0.51-0.94, P = 0.02) for N0 versus N1 tumors. The association between pRB loss and prognosis was examined in nine studies, and the summary hazard ratio was 1.39 (95% CI:1.11-1.74, P = 0.004). pRB inactive was significant associated with T3/T4 tumors and N1 stage as well as adverse prognosis for ESCCs. It appears warranted to prospectively validate that pRB loss may be used for subdividing the T,N stage evaluation of patients with ESCCs, and these patients may be the preponderant people for individualized treatment or target therapy.

Study on the safe range of shortening of the spinal cord in canine models.

STUDY DESIGN: Experimental dog model of spinal cord shortening. OBJECTIVES: To clarify the relationship between the amount of shortening of the spinal cord and the degree of injury it may induce, and to determine the safe range of the shortening. SETTING: Xi'an Jiaotong University, China. METHODS: Thirty adult dogs were randomly allocated to five groups. Dogs in Group A (sham operation control) underwent spondylectomy to have two-thirds of the thirteenth thoracic segment (T13) resected, without bone-to-bone contact of the adjacent vertebral bodies. Those in Group B, C, D and E had one-third, half, two-thirds and total of their T13 resected, respectively, with bone-to-bone contact. Somatosensory-evoked potentials (SEP) and spinal cord blood flow (SCBF) were detected. The histopathologic changes of spinal cord tissue were observed by hematoxylin and eosin stain and electron microscope. RESULTS: The shortening of the spinal cord < half of a vertebral segment height caused a reversible change of SEP. Whereas, the changes resulted from the shortening of more than two-thirds of a vertebral segment height did not return to the normal level. SCBF increased temporarily when the shortening was within two-thirds of a vertebral segment height; whereas, it decreased progressively when the length of the shortening was equal to one vertebral segment height. More serious hemorrhage occurred as the shortening increased. CONCLUSION: Shortening of half of a vertebral segment height will not induce spinal cord injury (SCI), while that between half and two-thirds of a vertebral segment may lead to incomplete SCI.

[Effect of lipopolysaccharide-stimulated exosomes from human dental pulp stem cells combined with stromal cell-derived factor-1 on dental pulp regeneration].

Objective: To investigate the effect of exosomes from mild-inflammation- stimulated human dental pulp stem cells (hDPSC) combined with stromal cell-derived factor-1 (SDF-1) on dental pulp regeneration in rats. Methods: Primary hDPSCs were isolated, cultured and then stimulated by using lipopolysaccharide (LPS). The exosomes from the hDPSCs with (L-EXO) or without (N-EXO) LPS were extracted by overspeed differential centrifugation and were identified by transmission electron microscopy and Western blotting. Forty SD rats, aged 6-8 weeks, were equally divided into S group (SDF-1 alone), L+S group (L-EXO combined with SDF-1), N+S group (N-EXO combined with SDF-1) and blank control group (no substance implanted into the root canal) by random number table method. Bilateral mandibular first molars were used as the experimental teeth to establish pulpless root canal models and different contents were implanted into the root canals according to the groups. All rats were over-anesthetized and sacrificed at the 30th day after content implantation. Bilateral mandibular tissues were taken for histological evaluation by means of HE, Masson and immunohistochemical stainings. Results: The HE staining showed new pulp-like tissue in the root canals of all three experimental groups. The amount of new tissues and the number of cells in the tissues were greatest in L+S group and least in S group. Masson staining showed that the mineralized tissue in L+S group was arranged longitudinally along the root canal wall and the collagen fibers were arranged in an orderly fashion, while those in N+S group showed an irregular and disordered distribution. Quantitative analysis of the area of neovascularization in each group showed that the density of vessels in the L+S group [(2.03±0.65)%] was significantly higher than that in the S group [(0.65±0.05)%] and the N+S group [(1.06±0.38)%] respectively (F=5.879, P<0.05). Immunohistochemical staining showed that the expression of CXC chemokine receptor 4 (CXCR4) was significantly lower in S and L+S groups than in N+S group, with a statistically significant difference (F=8.633, P<0.01). Conclusions: Exosomes secreted by hDPSCs combined with SDF-1 might increase the amount of new tissue in the root canal and the density of blood vessels in the tissue. L-EXO showed a stronger effect than N-EXO did. The combination of L-EXO with SDF-1 might result in more regular arrangement of mineralized tissue and collagen fibers in the regenerative tissues.

The current landscape of immunotherapy for pediatric brain tumors.

Pediatric central nervous system tumors are the most common solid malignancies in childhood, and aggressive therapy often leads to long-term sequelae in survivors, making these tumors challenging to treat. Immunotherapy has revolutionized prospects for many cancer types in adults, but the intrinsic complexity of treating pediatric patients and the scarcity of clinical studies of children to inform effective approaches have hampered the development of effective immunotherapies in pediatric settings. Here, we review recent advances and ongoing challenges in pediatric brain cancer immunotherapy, as well as considerations for efficient clinical translation of efficacious immunotherapies into pediatric settings.

Adult precision medicine: learning from the past to enhance the future.

Despite therapeutic advances for other malignancies, gliomas remain challenging solid tumors to treat. Complete surgical resection is nearly impossible due to gliomas' diffuse infiltrative nature, and treatment is hampered by restricted access to the tumors due to limited transport across the blood-brain barrier. Recent advances in genomic studies and next-generation sequencing techniques have led to a better understanding of gliomas and identification of potential aberrant signaling pathways. Targeting the specific genomic abnormalities via novel molecular therapies has opened a new avenue in the management of gliomas, with encouraging results in preclinical studies and early clinical trials. However, molecular characterization of gliomas revealed significant heterogeneity, which poses a challenge for targeted therapeutic approaches. In this context, leading neuro-oncology researchers and clinicians, industry innovators, and patient advocates convened at the inaugural annual Remission Summit held in Orlando, FL in February 2019 to discuss the latest advances in immunotherapy and precision medicine approaches for the treatment of adult and pediatric brain tumors and outline the unanswered questions, challenges, and opportunities that lay ahead for advancing the duration and quality of life for patients with brain tumors. Here, we provide historical context for precision medicine in other cancers, present emerging approaches for gliomas, discuss their limitations, and outline the steps necessary for future success. We focus on the advances in small molecule targeted therapy, as the use of immunotherapy as an emerging precision medicine modality for glioma treatment has recently been reviewed by our colleagues.