Bone metastasis in non-small-cell lung cancer: genomic characterization and exploration of potential targets.

Background: Bone metastasis (BM) seriously affects the quality of life and reduces the survival time of patients with non-small-cell lung cancer (NSCLC). The genomic characteristics and potential targets of BMs are yet to be fully explored. Objective: To explore the genetic characteristics and potential targets of BM in NSCLC. Design: In all, 83 patients with NSCLC were retrospectively selected in this study. Genomic characterization of BMs was explored with the analysis of NGS results from primary tumors and BMs in 6 patients, then combined with NGS results of lung tumors in 16 patients with initial recurrence in bone to analyze mutations potentially associated with BMs, and finally, the correlation was further validated in 61 postoperative patients. Methods: The next generation sequencing (NGS) was performed to identify genomic differences between pulmonary primary tumors and BM. Fluorescence in situ hybridization and immunohistochemistry were performed in postoperative tumor tissues from patients who had undergone radical surgery to validate the predictive role of molecular targets for BM. The correlation between cyclin-dependent kinase 4 (CDK4) and BM was evaluated by Pearson's chi-square test. The university of alabama at birminghan cancer data analysis portal (UALCAN) was carried out for the detection of CDK4 expression in lung cancer and the relationship between CDK4 and clinicopathological parameters. The relationship between prognosis and CDK4 expression was analyzed by the Kaplan-Meier plotter. Results: The rate of gene amplification was increased (24% versus 36%) while gene substitution/indel was decreased (64% versus 52%) in BMs. The BM-specific mutations were analyzed in 16 recurrent patients which revealed the highest incidence of CDK4 amplification (18.8%). According to the Kaplan-Meier plotter database, the NSCLC patients with high CDK4 gene expression showed poor overall survival (OS) and recurrence-free survival (RFS) (p < 0.05). The incidence of CDK4 amplification tended to be higher in recurrent patients compared to the patients without BM (18.8% versus 4.7%, p = 0.118). Conclusion: Compared to the primary tumors of NSCLC, the genome of BMs showed an increased proportion of amplification and a decreased proportion of gene substitution/indel. Furthermore, the CDK4 amplification ratio seemed to be elevated in NSCLC patients with BM which may be associated with poor OS and RFS.

Genomic characterization and potential targets of bone metastasis in non-small cell lung cancer NGS was performed on the matched primary tumors and bone metastases to explore the differences in the genomes of bone metastases, and it was found that gene amplification increased in bone metastases. Combined with the results of NGS in NSCLC patients with the first postoperative recurrence site in the bone, it was found that CDK4 amplification expression increased in bone metastases. Finally, the correlation between bone metastasis and CDK4 amplification was verified by expanding the sample.

Pulmonary salivary gland tumor-hyalinizing clear cell carcinoma: a literature review.

Primary pulmonary hyalinizing clear cell carcinoma (HCCC) is a very rare lung tumor that accounts for less than 0.09% of all primary lung tumors and has no specific epidemiology. The correct diagnosis requires imaging, laboratory, pathological, immunohistochemical, and molecular examination. The most typical feature of pulmonary HCCC is the clear cell component with clear stroma. In addition, the fusion gene EWSR1::ATF1 due to t(12;22)(q13;q12) is essential for the pathological diagnosis of pulmonary HCCC. The main treatment for pulmonary HCCC is surgery. This review focus on the pathological features, immunohistochemical examination, mutation analysis and treatment of pulmonary HCCC.

Codelivery of CuS and DOX into Deep Tumors with Size and Charge-Switchable PAMAM Dendrimers for Chemo-photothermal Therapy.

Accurate targeting of therapeutic agents to specific tumor tissues, especially via deep tumor penetration, has been an effective strategy in cancer treatments. Here, we described a flexible nanoplatform, pH-responsive zwitterionic acylsulfonamide betaine-functionalized fourth-generation PAMAM dendrimers (G4-AB), which presented multiple advantages for chemo-photothermal therapy, including template synthesis of ultrasmall copper sulfide (CuS) nanoparticles and further encapsulation of doxorubicin (DOX) (G4-AB-DOX/CuS), long-circulating performance by a relatively large size and zwitterionic surface in a physiological environment, combined size shrinkage, and charge conversions via pH-responsive behavior in an acidic tumor microenvironment (TME). Accordingly, high tumor penetration and positive cell uptake for CuS and DOX have been determined, which triggered an excellent combination treatment under near-infrared irradiation in comparison to the monochemotherapy system and irresponsive chemo-photothermal system. Our study represented great promise in constructing multifunctional carriers for the effective delivery of photothermal nanoparticles and drugs in chemo-photothermal therapy.

Beer-gut microbiome alliance: a discussion of beer-mediated immunomodulation via the gut microbiome.

As a long-established fermented beverage, beer is rich in many essential amino acids, vitamins, trace elements, and bioactive substances that are involved in the regulation of many human physiological functions. The polyphenols in the malt and hops of beer are also important active compounds that interact in both directions with the gut microbiome. This review summarizes the mechanisms by which polyphenols, fiber, and other beneficial components of beer are fermentatively broken down by the intestinal microbiome to initiate the mucosal immune barrier and thus participate in immune regulation. Beer degradation products have anti-inflammatory, anticoagulant, antioxidant, and glucolipid metabolism-modulating potential. We have categorized and summarized reported data on changes in disease indicators and in vivo gut microbiota abundance following alcoholic and non-alcoholic beer consumption. The positive effects of bioactive substances in beer in cancer prevention, reduction of cardiovascular events, and modulation of metabolic syndrome make it one of the candidates for microecological modulators.

Pickering emulsions stabilized by soybean protein isolate/chitosan hydrochloride complex and their applications in essential oil delivery.

In this work, fabrication of soybean protein isolate (SPI)/chitosan hydrochloride (CHC) composite particles stabilized O/W Pickering emulsions using soybean oil as an oil phase was optimized by examining the effects of pH, SPI/CHC mass ratio, SPI/CHC composite particle concentration and oil phase fraction on the stability of the emulsions. The results showed that under the conditions of SPI/CHC mass ratio 1:1, pH 4 and particle concentration 2 %, the SPI/CHC composite particles could stabilize the emulsions with oil phase fraction up to 80 %. At an oil phase fraction of 60 %, the emulsions had a minimum particle size. The microstructure, storage and oxidation stabilities and rheological properties of the emulsions were determined. Using this SPI/CHC composite particle-stabilized Pickering emulsion template, citrus essential oil (CEO) Pickering emulsion (CEOP) was prepared. CEOP was found to markedly inhibit two food-related microorganisms, Staphylococcus aureus and Escherichia coli. In addition, the CEOP emulsion dilution (containing 4500 µL CEO/L) not only improved the water solubility of CEO, but also effectively retarded the browning and bacterial growth of fresh-cut apple. The SPI/CHC-stabilized Pickering emulsion template constructed in this work provides a promising alternative for the delivery of antimicrobial essential oils in the food industry.

Soybean protein isolate/chitosan complex-rutin microcapsules.

Rutin is a flavonoid polyphenol with excellent biological activity, but due to its instability and poor water solubility, the utilization rate is reduced in vivo. Preparation of rutin microcapsules from soybean protein isolate (SPI) and chitosan hydrochloride (CHC) by composite coacervation can improve this restriction. The optimal preparation conditions were as follows: the volume ratio of CHC/SPI 1:8, pH 6, and total concentration of CHC and SPI 2 %. The rutin encapsulation rate and loading capacity of the microcapsules were 90.34 % and 0.51 % under optimal conditions. The SPI-CHC-rutin (SCR) microcapsules had a gel mesh structure and good thermal stability, and the system was stable and homogeneous after 12 d storage. During in vitro digestion, the release rates of SCR microcapsules in simulated gastric and intestinal fluids were 16.97 % and 76.53 %, respectively, achieving a targeted release of rutin in intestinal fluids; and the digested products were found to exhibit superior antioxidant activity to that of free rutin digests, indicating a good protection of microencapsulation on the bioactivity of rutin. Overall, SCR microcapsules developed in this study effectively enhanced the bioavailability of rutin. The present work provides a promising delivery system for natural compounds with low bioavailability and stability.

Synthesis of pH-responsive polyzwitterions for activated cellular uptake and tumor accumulation of gold nanoparticles at tumorous acidity.

The response sensitivity of surface material plays an important role in adjustable nano-bio interactionin vivo. In this present, a zwitterionic polymer (polyzwitterion) containing quaternary ammonium cation and sulfonamide anion poly(4-((4-(3-(methacryloyloxy)propoxy)phenyl) sulfonamido)-N, N, N-trimethyl-4-oxobutan-1-aminium chloride) (PMPTSA) was synthesized by Reversible Addition-Fragmentation Chain Transfer Polymerization (RAFT) polymerization to explore the pH responsive behavior in tumors. The PMPTSA-coated gold nanoparticles (PMPTSA-@-Au NPs) showed zwitterionic nature such as antifouling ability, low cellular uptake and prolonged circulation time similar with common hydrophilic polymers, including polyethylene glycol (PEG), poly(carboxybetaine methacrylate) and poly(sulfobetaine methacrylate) functional gold nanoparticles in physiological environment (pH 7.4). A high sensitivity and reversible positive charge conversion of P(MPTSA)-@-Au NPs at tumor slight acidic microenvironment (∼pH 6.8) leaded to an enhanced cellular internalization than that at pH 7.4 and increased tumor accumulation compared with PEG, polycarboxybetaines and polymer sulphobetaine (PSB) functional gold nanoparticles. The highly pH responsive PMPTSA will provide the promising application in cancer nanomedicine.

Pulmonary Salivary Gland Tumor, Mucoepidermoid Carcinoma: A Literature Review.

Pulmonary mucoepidermoid carcinoma (PMEC) is the most common malignant salivary gland tumor in the lungs and accounts for 0.1-0.2% of all lung malignancies in adults. It has no specific epidemiological or clinical characteristics. Correct diagnosis requires the combined examinations of images, laboratories, pathology, and immunohistochemistry (IHC) as well as molecular characteristics. PMEC tumors are characterized by squamous, intermediate, and mucus-secreting cells. Currently, histological appearance, mitotic frequency, cellular atypia, and necrocytosis allow the classification of PMEC into low grade or high grade. Molecular changes are crucial to pathological diagnosis. The driver of PMEC seems to be the fusion protein MECT1-MAML2 that is generated from a genetic mutation in t (11; 19) (q21; p13), while other gene mutations are also reported. However, no treatment of PMEC exists so far; surgical excision is still the primary treatment, while the efficacies of chemotherapy or radiotherapy are undefined. Tyrosine kinase inhibitor (TKI) therapy and immunotherapy showed to have significant therapeutic effects but require more investigation and better understanding. This review focuses on the clinical characteristics, imaging and pathologic features, immunohistochemical examination, mutation analysis, differential diagnosis, prognosis, and treatment of PMEC.

Research on the Multi-Element Synthetic Aperture Focusing Technique in Breast Ultrasound Imaging, Based on the Ring Array.

As a widely clinical detection method, ultrasonography (US) has been applied to the diagnosis of breast cancer. In this paper, the multi-element synthetic aperture focusing (M-SAF) is applied to the ring array of breast ultrasonography (US) imaging, which addresses the problem of low imaging quality due to the single active element for each emission and the reception in the synthetic aperture focusing. In order to determine the optimal sub-aperture size, the formula is derived for calculating the internal sound pressure of the ring array with a 200 mm diameter, and the sound pressure distribution is analyzed. The ring array with 1024 elements (1024 ring array) is established in COMSOL Multiphysics 5.6, and the optimal sub-aperture size is 16 elements, according to the sound field beam simulation and the directivity research. Based on the existing experimental conditions, the ring array with 256 elements (256 ring array) is simulated and verified by experiments. The simulation has a spatial resolution evaluation in the k-Wave toolbox, and the experiment uses nylon rope and breast model imaging. The results show that if the sub-aperture size has four elements, the imaging quality is the highest. Specifically, the spatial resolution is the best, and the sound pressure amplitude and signal-to-noise ratio (SNR) are maintained at a high level in the reconstructed image. The optimal sub-aperture theory is verified by the two kinds of ring arrays, which also provide a theoretical basis for the application of the multi-element synthetic aperture focusing technology (M-SAF) in ring arrays.

Synergistic size and charge conversions of functionalized PAMAM dendrimers under the acidic tumor microenvironment.

Developing nanomedicine with highly adaptive behaviors has shown great effectiveness in cancer treatment. However, the multi-functional integration of nano-therapeutic systems inevitably leads to complexity in the structure and impairs the operational efficiency or performance. Herein, we describe a novel nano-therapeutic system, G4-AB, capable of simultaneous dual conversions of the size and charge while targeting the acidic tumor microenvironment. G4-AB, containing a hydrophobic inner cavity for doxorubicin (DOX) loading, was synthesized by modifying amine-terminated 4th-generation polyamidoamine (G4-PAMAM) dendrimers with acylsulfonamide betaine (AB). Due to the dipole-dipole interaction among the AB moieties, G4-AB self-assembles to form micellar clusters with a zwitterionic surface. Possessing an anti-fouling property and suitable size, G4-AB exhibits optimized blood circulation under physiological pH conditions. Moreover, the extracellular pH value of the tumor microenvironment (pH 6.5) can trigger the protonation of acylsulfonamide, resulting in the cationization of AB and dissociation of G4-AB into unimolecular micelles (∼12 nm) due to electrostatic repulsion. The synergistic dual conversions further ensure drug accumulation with enhanced tumor penetration and cell internalization. The in vitro and in vivo experiments demonstrate that the G4-AB-DOX nano-therapeutic system possesses better antitumor efficiency and lower toxicity than free DOX or PEGylated PAMAM.

The activated DNA double-strand break repair pathway in cumulus cells from aging patients may be used as a convincing predictor of poor outcomes after in vitro fertilization-embryo transfer treatment.

Women with advanced maternal age exhibit low anti-Müllerian hormone (AMH) levels and an altered follicular environment, which is associated with poor oocyte quality and embryonic developmental potential. However, the underlying mechanism is poorly understood. The present study aimed to assesswhether aging patients exhibit an activated DNA double-strandbreak (DSB) repair pathway in cumulus cells and thus, an association with poor outcomes after in vitro fertilization-embryo transfer (IVF-ET) treatment. Cumulus cells from young (≤29 y) and aging (≥37 y) human female patients were collected after oocyte retrieval. Our results indicated that aging patients showed a higher rate of γ-H2AX-positive cells than in young patients (24.33±4.55 vs.12.40±2.31, P<0.05). We also found that the mRNA expression levels of BRCA1, ATM, MRE11 and RAD51 were significantly elevated in aging cumulus cells. Accordingly, significantly increased protein levels of phospho-H2AX, BRCA1, ATM, MRE11 and RAD51 could be observed in aging cumulus cells. Moreover, aging cumulus cells showed a more frequent occurrence of early apoptosis than young cumulus cells. This study found that increases in DSBs and the activation of the repair pathway are potential indicators that may be used to predictoutcomes after IVF-ET treatment.

Heterologous expression of carcinoembryonic antigen in Lactococcus lactis via LcsB-mediated surface displaying system for oral vaccine development.

BACKGROUND/PURPOSE: Carcinoembryonic antigen (CEA) is an attractive target for immunotherapy because it is expressed minimally in normal tissue, but is overexpressed in a wide variety of malignant epithelial tissues. Lactic acid bacteria (LABs), widely used in food processes, are attractive candidates for oral vaccination. Thus, we examined whether LABs could be used as a live vaccine vector to deliver CEA antigen. METHODS: CEA was cloned into an Escherichia coli/Lactococcus lactis shuttle vector pSEC:LEISS under the control of a nisin promoter. For displaying the CEA on the cell surface of the L. lactis strain, the anchor motif LcsB from the S-layer protein of Lactobacillus crispatus was fused with CEA. Intracellular and cell surface expression of the CEA-LcsB fusion was confirmed by western blot analysis. RESULTS: Significantly higher levels of CEA-specific secretory immunoglobulin A in the sera of mice were observed upon oral administration of strain cultures containing the CEA-LcsB fused protein. In addition, the CEA-LcsB antigen group showed a higher spleen index compared to the CEA antigen alone or negative control, demonstrating that surface-displayed CEA antigen could induce a higher immune response. CONCLUSION: These results provided the first evidence for displaying CEA antigen on the cell surfaces of LABs as oral vaccines against cancer or infectious diseases.

In vitro evaluation of Lactobacillus crispatus K313 and K243: high-adhesion activity and anti-inflammatory effect on Salmonella braenderup infected intestinal epithelial cell.

Currently, there is an increasing interest in the use of probiotics as an alternative strategy to antimicrobial compounds. In this study, two high adhesive strains Lactobacillus crispatus K313 adhering to HT-29 cells as well as Lb. crispatus K243 adhering to collagen type IV were isolated from chicken intestines. SDS-PAGE analysis revealed the presence of the potential S-proteins SlpA and SlpB in Lb. crispatus K243 and K313. SlpA and SlpB, rich in hydrophobic amino acids, were proved to be involved in adhering to collagen type IV and HT-29 cells, respectively, based on the LiCl treatment assay. After removal of S-proteins, the viability and tolerance of the two Lb. crispatus strains to simulated gastric and small intestinal juice were reduced, indicating the protective role of S-proteins against the hostile environments. Lb. crispatus K313 exhibited the stronger autoaggregation ability and inhibitive activity against Salmonella braenderup H9812 adhesion to HT-29 cells than the strain K243. To elucidate the inhibitive mechanism, cultured epithelial cells were exposed with Lb. crispatus strains, and followed by a challenge with S. braenderup H9812. The pro-inflammatory signaling factors (IL-8, CXCL1 and CCL20) from HT-29 were detected by real-time PCR technology. The results showed that both of Lb. crispatus strains down-regulated the transcription level of those pro-inflammatory genes induced by S. braenderup H9812 by 36.2-58.8%. ELISA analysis was further confirmed that Lb. crispatus K243 and K313 inhibited the IL-8 secretion triggered by S. braenderup H9812 by 32.8% and 47.0%, indicating that the two isolates could attenuate the pro-inflammatory signaling induced by S. braenderup H9812, and have the potential application in clinical practice to prevent diarrhea.

[Pregnancy outcomes of in vitro fertilization and embryo transfer in infertile women with polycystic ovarian syndrome].

OBJECTIVE: To investigate the pregnancy outcomes of in vitro fertilization (IVF) and embryo transfer (ET) in infertile women with polycystic ovarian syndrome (PCOS). METHODS: From August 2005 to June 2011, 200 IVF-ET cycles performed in women with polycystic ovarian syndrome in Shengjing Hospital and Shenyang 204 Hospital were enrolled in this retrospective study, matched with 400 IVF-ET cycles in infertile women with fallopian tube factors as control group. The incidence of abortion, gestational diabetes mellitus (GDM), hypertensive disorders in pregnancy, preterm birth, small for gestational age infant (SGA), large for gestational age infant (LGA), neonatal asphyxia, neonatal death and deformity was compared between two groups. RESULTS: The incidence of spontaneous abortion was 26.0% (52/200) in PCOS group, which was significantly higher than 10.2% (41/400) in control group (P < 0.05). The incidence of GDM, hypertensive disorders in pregnancy, preterm birth, cesarean section in PCOS group was 23.6% (35/148), 16.2% (24/148), 17.6% (26/148), 83.1% (123/148), which were significantly higher than 4.2% (15/359), 6.1% (22/359), 7.8% (28/359), 73.8% (265/359) in control group (P < 0.05). The incidence of SGA, LGA, neonatal asphyxia, neonatal death and deformity did not show remarkable difference between two groups, which were 2.7% (4/148), 4.7% (7/148), 5.4% (8/148), 0 in PCOS group and 1.4% (5/359), 2.2% (8/359), 2.8% (10/359), 0 in control group (P > 0.05). CONCLUSION: IVF-ET is an effective treatment for infertile women with PCOS, however, the incidence of spontaneous abortion, GDM, hypertensive disorders in pregnancy, preterm birth, cesarean section in PCOS patients was increased.