Fate mapping of Spp1 expression reveals age-dependent plasticity of disease-associated microglia-like cells after brain injury.

Microglial reactivity to injury and disease is emerging as a heterogeneous, dynamic, and crucial determinant in neurological disorders. However, the plasticity and fate of disease-associated microglia (DAM) remain largely unknown. We established a lineage tracing system, leveraging the expression dynamics of secreted phosphoprotein 1（Spp1） to label and track DAM-like microglia during brain injury and recovery. Fate mapping of Spp1+ microglia during stroke in juvenile mice revealed an irreversible state of DAM-like microglia that were ultimately eliminated from the injured brain. By contrast, DAM-like microglia in the neonatal stroke models exhibited high plasticity, regaining a homeostatic signature and integrating into the microglial network after recovery. Furthermore, neonatal injury had a lasting impact on microglia, rendering them intrinsically sensitized to subsequent immune challenges. Therefore, our findings highlight the plasticity and innate immune memory of neonatal microglia, shedding light on the fate of DAM-like microglia in various neuropathological conditions.

Retinoid X receptor gamma dictates the activation threshold of group 2 innate lymphoid cells and limits type 2 inflammation in the small intestine.

Group 2 innate lymphoid cells (ILC2s) are crucial in promoting type 2 inflammation that contributes to both anti-parasite immunity and allergic diseases. However, the molecular checkpoints in ILC2s that determine whether to immediately launch a proinflammatory response are unknown. Here, we found that retinoid X receptor gamma (Rxrg) was highly expressed in small intestinal ILC2s and rapidly suppressed by alarmin cytokines. Genetic deletion of Rxrg did not impact ILC2 development but facilitated ILC2 responses and the tissue inflammation induced by alarmins. Mechanistically, RXRγ maintained the expression of its target genes that support intracellular cholesterol efflux, which in turn reduce ILC2 proliferation. Furthermore, RXRγ expression prevented ILC2 response to mild stimulations, including low doses of alarmin cytokine and mechanical skin injury. Together, we propose that RXRγ expression and its mediated lipid metabolic states function as a cell-intrinsic checkpoint that confers the threshold of ILC2 activation in the small intestine.

Dopamine inhibits group 2 innate lymphoid cell-driven allergic lung inflammation by dampening mitochondrial activity.

Neuronal signals have emerged as pivotal regulators of group 2 innate lymphoid cells (ILC2s) that regulate tissue homeostasis and allergic inflammation. The molecular pathways underlying the neuronal regulation of ILC2 responses in lungs remain to be fully elucidated. Here, we found that the abundance of neurotransmitter dopamine was negatively correlated with circulating ILC2 numbers and positively associated with pulmonary function in humans. Dopamine potently suppressed lung ILC2 responses in a DRD1-receptor-dependent manner. Genetic deletion of Drd1 or local ablation of dopaminergic neurons augmented ILC2 responses and allergic lung inflammation. Transcriptome and metabolic analyses revealed that dopamine impaired the mitochondrial oxidative phosphorylation (OXPHOS) pathway in ILC2s. Augmentation of OXPHOS activity with oltipraz antagonized the inhibitory effect of dopamine. Local administration of dopamine alleviated allergen-induced ILC2 responses and airway inflammation. These findings demonstrate that dopamine represents an inhibitory regulator of ILC2 responses in allergic airway inflammation.

Intranuclear Delivery of DNA Nanostructures via Cellular Mechanotransduction.

DNA nanostructures are attractive gene carriers for nanomedicine applications, yet their delivery to the nucleus remains inefficient. We present the application of extracellular mechanical stimuli to activate cellular mechanotransduction for boosting the intranuclear delivery of DNA nanostructures. Treating mammalian cells with polythymidine-rich spherical nucleic acids (poly(T) SNAs) under gentle compression by a single coverslip leads to up to ∼50% nuclear accumulation without severe endosomal entrapment, cytotoxicity, or long-term membrane damage; no chemical modification or transfection reagent is needed. Gentle compression activates Rho-ROCK mechanotransduction and causes nuclear translocation of YAP. Joint compression and treatment with poly(T) oligonucleotides upregulate genes linked to myosin, actin filament, and nuclear import. In turn, Rho-ROCK, myosin, and importin mediate the nuclear entry of poly(T) SNAs. Treatment of endothelioma cells with poly(T) SNAs bearing antisense oligonucleotides under compression inhibits an intranuclear oncogene. Our data should inspire the marriage of DNA nanotechnology and cellular biomechanics for intranuclear applications.

A pH-Reversible Fluorescent Probe for in Situ Imaging of Extracellular Vesicles and Their Secretion from Living Cells.

Our knowledge in how extracellular vesicles (EVs) are secreted from cells remains inadequate due to the limited technologies available for visualizing them in situ. We report a pH-reversible boron dipyrromethene (BODIPY) fluorescent probe for confocal imaging of EVs secreted from living cells without inducing severe cytotoxicity. This probe predominantly assumes a non-fluorescent leuco-BODIPY form under basic conditions, but it gradually switches to its fluorescent parent BODIPY form upon acidification; such pH transition empowers the imaging of acidic EVs (such as CD81-enriched exosomes and extracellular multivesicular bodies) in weakly basic culture medium and intracellular acidic precursor EVs in weakly basic cytoplasm, with minimal false positive signals frequently encountered for "always-on" dyes. Joint application of this probe with plasmid transfection reveals the secretion of some EVs from cellular pseudopodia via microtubule trackways. This probe may provide mechanistic insights into the extracellular transport of EVs and support the development of EV-based nanomedicines.

Sub-10 nm Substrate Roughness Promotes the Cellular Uptake of Nanoparticles by Upregulating Endocytosis-Related Genes.

Nanosubstrate engineering is an established approach for modulating cellular responses, but it remains infrequently exploited to facilitate the intracellular delivery of nanoparticles (NPs). We report nanoscale roughness of the extracellular environment as a critical parameter for regulating the cellular uptake of NPs. After seeding cells atop a substrate that contains randomly immobilized gold NPs (termed AuNP-S) with sub-10 nm surface roughness, we demonstrate that such cells internalize up to ∼100-fold more poly(ethylene glycol)-coated AuNPs (Au@PEG NPs) than those cells seeded on a conventional flat culture plate. Our result is generalizable to 4 different cell types and Au@PEG NPs modified with 13 different hydrocarbyl functional groups. Conditioning cells to AuNP-S not only leads to upregulation of clathrin- and integrin-related genes, but also supports elevated uptake of Au@PEG NPs via clathrin-mediated endocytosis. Our data suggest a simple and robust method for boosting the intracellular delivery of nanomedicines by nanosubstrate engineering.

Clinical Research of the Remanent Diplopia Correction Operation After Orbital Fracture Repair.

OBJECTIVE: To investigate the occasion, method and effectiveness of the diplopic correction after the orbital fracture repair in Chinese patients with orbital fracture. METHODS: This was a retrospective observational study of 64 patients with remanent diplopia after the reconstructions of orbital fracture were studied between January 2014 and June 2017. The different cause, operational occasions, and surgical techniques analyzed. All patients were followed for 6 months after the surgeries. RESULTS: After treatment, 64 patients with III degree diplopia were all performed the operations on extraocular muscles. Different operation was selected to different patients according to the examinational results of the extraocular muscles and passive drawer test, including rectos retropulsion, suspension, excise, Jensen's connection, and fixation to periorbital membrane. During the surgery, 25 patients had no diplopia in mainly functional visual fields, while 15 patients still had III degree diplopia. After 6 months, 27 patients had no diplopia in mainly functional visual fields, while 13 patients still had II degree diplopia. Among these 13 patients with III degree diplopia, 7 patients had no the diplopia of primary position of eye, and 6 patients had the diplopia of primary position. These 6 patients were suggested to wear the triangular prisms for improvement of the diplopia. CONCLUSION: The operations should be selected according to the examining results about eye movements and diplopia before and during the surgeries. Reasonable operations could correct diplopia and improve the eye movements effectually.

Comprehensive Evaluation of Visual Function in Patients With Orbital Fracture by Visual Electrophysiology and Visual Field Examination.

OBJECTIVE: At present, the effect of the visual electrophysiology and vision field examination in patients with orbital blowout fracture is rarely studied. So, the authors investigate the value of visual electrophysiology and vision field examination in the diagnosis of ocular contusion. METHODS: The position and range of fracture of 81 patients were determined by computed tomography (CT) scanning. Visual evoked potential (VEP), electroretinogram (ERG), and mfERG were vision field examination detected in 81 patients and the results were compared with those of contralateral healthy eyes. In addition, visual electrophysiology and vision field examination in diagnosis of eye contusion was analyzed and the correlation of the VEP, ERG, mfERG injury duration, and visual acuity was further analyzed. RESULTS: The visual acuity of orbital fractures was significantly decreased compared with that in the uninjured eyes (t = 2.181, P = 0.032). Compared injured eyes and normal eyes in 54 patients, b wave of Max-ERG and Cone-ERG implied value extension (t = -2.426, P = 0.025; t = -2.942, P = 0.014), P-VEP P100 Peak duration and amplitude significantly extended (t = 3.162, P = 0.007; t = 9.314, P = 0.000), and F-VEP P1 amplitude decreased significantly (t = 3.362, P = 0.004). mfERG showed that the injured eye central reaction was significantly decreased (t = 8.727, P = 0.000). There was a significant correlation between P-VEP P100 amplitude and visual acuity (r = 0.067, P = 0.000). But there was no significant correlation between the P100 peak value, amplitude of P-VEP, mfERG central reaction, and injured days, respectively. There was significant difference between 2 groups with average visual acuity and mean defect value (t = 3.253, 3.461, P = 0.006, 0.003). There was statistical means the difference in P-VEP abnormal group, visual field abnormal group, and combined detection abnormal groups, the abnormal rate increased significantly (χ = 3.931, P < 0.01). CONCLUSION: Orbital floor fracture can lead to optic nerve damage and also may be associated with decreased macular function. The combination analysis of visual electrophysiology and vision field examination is beneficial to early diagnosis of ocular trauma and can improve the positive rate in clinic practice.

Postoperative Improvement of Diplopia and Extraocular Muscle Movement in Patients With Reconstructive Surgeries for Orbital Floor Fractures.

OBJECTIVE: With orbital floor fracture incidence rates increasing year by year, many patients require surgical treatment to improve diplopia, limitation of extraocular muscle movement (EOM), enophthalmos, and midface appearance. With the use of high-density polyethylene, titanium screws, titanium plate, and titanium mesh to repair an orbital floor fracture, enophthalmos and midfacial deformity correction procedures have made great progress. However, attenuating diplopia and the limitation of EOM are still difficult problems to prevent. METHODS: The clinical data of 92 patients with reconstructive surgeries after orbital floor fracture were prospectively studied. The position, range of fracture, and incarceration of extraocular muscles were determined by computed tomography scanning. A Hess screen and a synoptophore examination were used to determine the EOM and diplopia. The Hess area ratio (HAR%) and the grade of diplopia were measured before and 1, 3, and 6 months after surgery. Diplopia was evaluated, and the severity was recorded accordingly. RESULTS: Diplopia rates in patients with simple orbital floor fracture showed a significant difference preoperatively and postoperatively at 1 and 3 months (P < 0.05) compared with diplopia rates at 6 months and 12 months after operation, which had no significant difference (P > 0.05). There was a statistically significant difference (P < 0. 05) among patients with an orbital floor fracture who had an operation within 3 weeks or more than 3 months after injury. Compared with preoperative and postoperative findings at 1 and 3 months, the limitation of EOM in patients with orbital floor fractures had a significant difference (P < 0.05); however, compared with 3 and 6 months after operation, there was no significant difference (P > 0.05). There was a statistically significant difference (P < 0.05) in patients with orbital floor fractures having had an operation within 3 weeks and more than 3 months after injury. The difference of improvement for diplopia after operation among HAR% < 65%, 65% ≤ HAR% ≤ 85%, and HAR% >85% groups were statistically significant (P <0.05). Postsurgically, the HAR% of the patients was improved, and the difference of HAR% between patients before and after the operation was statistically significant (P < 0.05). CONCLUSION: Surgical management can effectively improve diplopia and EOM disorder of patients with orbital floor fractures. Improvement statistics were calculated by a Hess screen and a synoptophore. Hess area ratio is a useful method to convert the Hess screen into a numerical value and can therefore be used to compare patients in clinical treatment of orbital wall fracture.

Antibody Fc-receptor FcεR1γ stabilizes cell surface receptors in group 3 innate lymphoid cells and promotes anti-infection immunity.

Group 3 innate lymphoid cells (ILC3) are crucial for maintaining mucosal homeostasis and regulating inflammatory diseases, but the molecular mechanisms governing their phenotype and function are not fully understood. Here, we show that ILC3s highly express Fcer1g gene, which encodes the antibody Fc-receptor common gamma chain, FcεR1γ. Genetic perturbation of FcεR1γ leads to the absence of critical cell membrane receptors NKp46 and CD16 in ILC3s. Alanine scanning mutagenesis identifies two residues in FcεR1γ that stabilize its binding partners. FcεR1γ expression in ILC3s is essential for effective protective immunity against bacterial and fungal infections. Mechanistically, FcεR1γ influences the transcriptional state and proinflammatory cytokine production of ILC3s, relying on the CD16-FcεR1γ signaling pathway. In summary, our findings highlight the significance of FcεR1γ as an adapter protein that stabilizes cell membrane partners in ILC3s and promotes anti-infection immunity.

Neuromedin U programs eosinophils to promote mucosal immunity of the small intestine.

Eosinophils are granulocytes that play an essential role in type 2 immunity and regulate multiple homeostatic processes in the small intestine (SI). However, the signals that regulate eosinophil activity in the SI at steady state remain poorly understood. Through transcriptome profiling of eosinophils from various mouse tissues, we found that a subset of SI eosinophils expressed neuromedin U (NMU) receptor 1 (NMUR1). Fate-mapping analyses showed that NMUR1 expression in SI eosinophils was programmed by the local microenvironment and further enhanced by inflammation. Genetic perturbation and eosinophil-organoid coculture experiments revealed that NMU-mediated eosinophil activation promotes goblet cell differentiation. Thus, NMU regulates epithelial cell differentiation and barrier immunity by stimulating NMUR1-expressing eosinophils in the SI, which highlights the importance of neuroimmune-epithelial cross-talk in maintaining tissue homeostasis.

Anti-proliferation effects of Sirolimus sustained delivery film in rabbit glaucoma filtration surgery.

PURPOSE: To investigate the efficacy, safety, and mechanisms of Sirolimus sustained delivery film on prevention of scar formation in a rabbit model of glaucoma filtration surgery. METHODS: Sixty-four New Zealand white rabbits who underwent trabeculectomy in the right eye were randomly allocated to one of the four treatment regimens: Sirolimus sustained delivery film treatment group (Group A), or drug-free film treatment group (Group B), or 30 ng/ml Sirolimus-soaked sponge treatment group (Group C), or no adjunctive treatment group (Group D), and each group consists of 16 rabbits. Intraocular pressure (IOP), morphologic changes of bleb, anterior chamber flare, and corneal endothelial cell count and complications were evaluated over a 28-day period follow-up time. Aqueous humor samples were gathered from Group A, and the concentration of Sirolimus was measured regularly post-operation. Rabbits were sacrificed on the 7th, 14th, and 28th day post-operation separately, and the fibroblast hypertrophy, infiltration of inflammatory, and proliferation of new collagen fiber formation in each group were evaluated with HE and Masson staining. Proliferative cell nuclear antigen (PCNA) and fibroblast apoptosis were evaluated by immunohistochemistry and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) assay at the 28th day post-operation. RESULTS: Both Sirolimus sustained delivery film (Group A) and Sirolimus alone (Group C) were well tolerated in this model, and significantly prolonged bleb survival compared with no drug treatment group (Group B and D; p<0.001). Group A had the longest bleb survival time in comparison with other groups (p<0.001). There were significant differences in IOP readings between Group A and other groups at the last follow-up (p<0.05). The concentration of Group A maintained stable for over 2 weeks, drops from (10.56 ±0.05) ng/ml at day 3 to (7.74 ±0.05) ng/ml at day 14. The number of corneal endothelial cells of Group A was not statistically significant between pre and post-operation. Histologic examination demonstrated that eyes treated with Sirolimus, especially the Sirolimus sustained delivery film, showed an obvious reduction in subconjunctival fibroblast scar tissue formation compared with no drug treatment groups, and had minimal evidence of inflammatory cell infiltration and new collagen deposition in the subconjunctiva. Immunohistochemistry assay showed that PCNA-expression was lower in the Group A (16.25±3.24%) compared to other groups (p<0.01). TUNEL assay showed a significant increase in the number of apoptotic fibroblasts around the surgical area in Group A and Group C (9.75±1.71% and 8.50±1.92%) compared to the Group B and D (p<0.01). CONCLUSIONS: Sirolimus drug sustained delivery film can inhibit inflammatory cell activity, impede fibroblast proliferation activity, and induce fibroblast apoptosis in the filtration surgery sites in rabbit. The results indicate a safe and effective treatment strategy in anti-scaring treatment in glaucoma surgery.

[Long-term effects of non-penetrating trabecular surgery versus trabeculectomy for treating glaucoma].

OBJECTIVE: To evaluate clinical outcomes of non-penetrating trabecular surgery (NPTS) and trabeculectomy surgery (TS) in the treatment of primary open angle glaucoma (POAG). METHODS: It was a case-control study. A total of 63 patients (63 eyes) with POAG were observed retrospectively. Thirty one eyes and 32 eyes underwent NPTS and TS, respectively. Intraocular pressure (IOP), filtration bleb, visual field and post-operative complications were observed for 6-60 months. The CMH χ(2) test was used to analyse the difference of them. RESULTS: After operation, the IOP in the NPTS group were from (13.87 ± 4.88) mm Hg (1 mm Hg = 0.133 kPa) to (24.01 ± 6.55) mm Hg, the IOP in the TS group were from (11.90 ± 4.92) mm Hg to (19.10 ± 7.43) mm Hg. The IOP in the NPTS group was significantly higher than that in the TS group (F = 5.137, P < 0.05). The ratio of sustained filtration bleb of NPTS group after surgery was 25/31 (80.6%), while 6/31 were flat filtration bleb. There were statistically significant difference in the rate of disappearance of filtration bleb between these two groups (χ(2) = 8.129, P < 0.05). The difference of visual field loss postoperatively between these two groups was not statistically significant. The incidence rate of newly developed cataract after NPTS and TS was 6/31 and 12/32, respectively. The difference of rate of complication between these two groups was statistically non-significant (χ(2) = 3.797, P < 0.05). The successful rate after NPTS and TS was 61.54% (16/26) and 14.29% (4/28), respectively. The difference of successful rate between these two groups was statistically significant (χ(2) = 14.463, P < 0.05). CONCLUSIONS: Both NPTS and TS are effective methods for the treatment of POAG. Postoperative complications after NPTS are less than those of TS, But patients with TS could maintain a lower IOP than those with NPTS. Long-term efficacy of NPTS is uncertain, it's important to choose the suitable surgery to gain a high success rate.

Biofilm microenvironment activated supramolecular nanoparticles for enhanced photodynamic therapy of bacterial keratitis.

Infectious keratitis caused by bacterial biofilms is one of the main causes of corneal blindness, presenting a serious threat to public health. In this study, matrix metalloproteinase (MMP)-sensitive supramolecular nanoparticles (denoted as MMP-S NPs) were constructed for enhancing photodynamic antibacterial effect against biofilm-associated bacterial keratitis. MMP-S NPs were prepared by host-guest self-assembly of chlorin e6 (Ce6) conjugated ß-cyclodextrin (ß-CD) prodrug (ß-CD-Ce6) and MMP-9-sensitive peptides (YGRKKKRRQRRR-GPLGVRG-EEEEEE) terminated with adamantane (Ad) (Ad-MMP-S PEPs). MMP-S NPs with EEEEEE peptide shell had a negatively charged surface, preventing adhesion to the normal ocular surface or healthy corneal cells, thus enhancing tear retention time. After arriving at the infected lesions, the protective EEEEEE peptide shell of MMP-S NPs was removed, triggered by overexpressed MMP-9 in the keratitis microenvironment. The subsequently exposed cationic peptides helped the nanoparticles penetrate and accumulate in biofilms as well as bind to Gram-negative bacteria Pseudomonas aeruginosa (P. aeruginosa), which eventually improved the photodynamic antibacterial effect. Furthermore, the P. aeruginosa keratitis model verified the high effectiveness of a topical eye drop formulation of MMP-S NPs in killing bacteria by destroying the bacterial membrane as a result of in situ photodynamic activation of reactive oxygen species (ROS) formation under light irradiation. Moreover, the inflammatory response in the cornea was inhibited to a great extent. As a result, further damage to the corneal tissue was completely suppressed. This research provides a viable antibacterial alternative to fight against bacterial keratitis through effective elimination of infectious bacteria and eradication of bacterial biofilms in the cornea.

Combination of hesperetin and platinum enhances anticancer effect on lung adenocarcinoma.

Lung cancer remains the leading cause of oncological death. There is an urgent need to discover new molecular targets and to develop new treatments. One of the UDP-glucuronosyltransferases (UGTs) family, UGT1A3, is highly expressed in lung adenocarcinoma (LUAD), and is associated with poor prognosis. Its inhibitor, hesperetin, may play an important role in anticancer therapy. The purpose of this study was to investigate the role of UGT1A3 in the progression of lung adenocarcinoma and to explore the value of its inhibitor hesperetin in the treatment of LUAD. Hesperetin suppressed lung adenocarcinoma cell proliferation and migration. The combination treatment of hesperetin with platinum suppressed tumor progression more significantly, especially compared with single drug treatment. UGT1A3 is an important prognostic factor for LUAD, and hesperetin can synergize platinum drugs by inhibiting UGT1A3 and increasing levels of reactive oxygen species (ROS).

Mining and validating the expression pattern and prognostic value of acetylcholine receptors in non-small cell lung cancer.

Acetylcholine receptors (AChRs), including nicotinic acetylcholine receptors (nAChRs) and muscarinic acetylcholine receptors (mAChRs), are highly expressed in bronchial epithelial cells.We used The Cancer Genome Atlas (TCGA) data set to evaluate the expression pattern and prognostic value of the AChR gene family in non-small cell lung cancer (NSCLC). The mined data was validated by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC).The survival analysis of TCGA data set showed that only CHRNA7 in the AChR gene family affected prognosis in both lung adenocarcinoma and lung squamous cell carcinoma. Furthermore, qRT-PCR proved that CHRNA7 was significantly upregulated in tumor tissues compared with matched normal tissues at mRNA level (P = .001). The expression level of α7 nAChR (encoded by CHRNA7) in 141 patients was measured by IHC and a high expression of α7 nAChR was associated with unfavorable prognosis (P = .008). Multivariate analysis showed that α7 nAChR was an independent prognostic factor (HR = 2.041; 95% CI 1.188-3.506; P = .007).α7 nAChR was upregulated in NSCLC and was associated with unfavorable prognosis. This gene may be a potential target for lung cancer treatment.

Tripartite motif-containing 15 overexpression in non-small cell lung cancer is associated with poor patient prognoses.

Purpose: This study aimed to comprehensively investigate the differential expression and prognostic indicators of the tripartite motif-containing (TRIM) gene family in non-small cell lung cancer (NSCLC). Methods: The Cancer Genome Atlas (TCGA) Research Network and three datasets from Gene Expression Omnibus (GEO) database were used to assess TRIM gene family expression patterns in NSCLC. Quantitative real-time PCR and immunohistochemistry (IHC) were conducted to confirm differentially expressed genes (DEGs). Kaplan-Meier survival analysis and univariate Cox regression analysis were carried out to analyze the association between TRIM gene expression and NSCLC prognoses. Gene set enrichment analysis (GSEA) was carried on for the predict the biological processes. Results: Of the 78 TRIM family members measured, TRIM15 was selected due to the DEGs and the prognostic value regarding NSCLC. In lung squamous cell carcinoma (LUSC), the Log2 fold change (Log2FC) of TRIM15 was 5.16 (p= 0.00575), whereas in lung adenocarcinoma (LUAD), it was 6.37 (p =6.78E-07). TRIM15 upregulation was related to poor prognoses in both LUSC (HR 1.353; 95%CI 1.023-1.789; p =0.034) and LUAD (HR 1.560; 95%CI 1.159-2.101; p =0.003). Using immunohistochemistry, TRIM15 expression was significantly higher in NSCLC tissues compared with that of matched normal tissues (p =0.0009), and similar findings were generated with tissue microarray analysis (p<0.0001). Conclusion: TRIM15 could act as a diagnostic predictor or therapeutic target for lung cancer treatments.

Intrapulmonary Cellular-Level Distribution of Inhaled Nanoparticles with Defined Functional Groups and Its Correlations with Protein Corona and Inflammatory Response.

Concerns over the health risks associated with airborne exposure to ultrafine particles [PM0.1, or nanoparticles (NPs)] call for a comprehensive understanding in the interactions of inhaled NPs along their respiratory journey. We prepare a collection of polyethylene glycol-coated gold nanoparticles that bear defined functional groups commonly identified in atmospheric particulates (Au@PEG-X NPs, where X = OCH3, COOH, NH2, OH, or C12H25). Regardless of the functional group, these ∼50 nm NPs remain colloidally stable following aerosolization and incubation in bronchoalveolar lavage fluid (BALF), without pronouncedly crossing the air-blood barrier. The type of BALF proteins adhered onto the NPs is similar, but the composition of protein corona depends on functional group. By subjecting Balb/c mice to inhalation of Au@PEG-X NPs for 6 h, we demonstrate that the intrapulmonary distribution of NPs among the various types of cells (both found in BALF and isolated from the lavaged lung) and the acute inflammatory responses induced by inhalation are sensitive to the functional group of NPs and postinhalation period (0, 24, or 48 h). By evaluating the pairwise correlations between the three variables of "lung-nano" interactions (protein corona, intrapulmonary cellular-level distribution, and inflammatory response), we reveal strong statistical correlations between the (1) fractions of albumin or carbonyl reductase bound to NPs, (2) associations of inhaled NPs to neutrophils in BALF or macrophages in the lavaged lung, and (3) level of total protein in BALF. Our results provide insights into the effect of functional group on lung-nano interactions and health risks associated with inhalation of PM0.1.

Expression profiling of ubiquitin-related genes in LKB1 mutant lung adenocarcinoma.

Liver kinase B1 (LKB1) is a tumor suppressor, and there is a very high proportion of LKB1 mutation in lung adenocarcinoma. The function of LKB1 is closely related to that of ubiquitin related genes. Our objective is to analyze the changes in ubiquitin-related genes in LKB1 mutant lung adenocarcinoma. We searched The Cancer Genome Atlas (TCGA) and obtained gene expression profiles from 230 lung adenocarcinoma patients, which were then analyzed using R software. Kaplan-Meier curves and Cox proportional hazards regression were applied to estimate survival. Real-time reverse transcription PCR was used to verify gene expression. Gene function was explored by gene set enrichment analysis. There were significantly expressed differences in the ubiquitin-related gene SH3RF1 between the LKB1 mutant and wild-type lung adenocarcinoma patients (p = 9.78013E-05). Patients with LKB1 mutation and high expression of SH3RF1 had a better prognosis than the low expression group (HR 0.356, 95% CI 0.136-0.929, p = 0.035). SH3RF1 can influence cell cycle, apoptosis, DNA replication and the p53 signaling pathway. SH3RF1 might have great clinical value act as a diagnostic biomarker and indicator to evaluate the prognosis of LKB1 mutant lung adenocarcinoma patients. This gene also can become a new treatment target for LKB1 mutant lung adenocarcinoma patients.