CircSTRBP contributes to H2O2-induced lens epithelium cell dysfunction through increasing NOX4 mRNA stability by recruiting IGF2BP1.

Previous studies have shown that the development of age-related cataract (ARC) is involved in lens epithelium dysfunction, which is associated with abnormally expressed circular RNAs (circRNAs). The current work aims to probe the role of circSTRBP (hsa_circ_0088,427) in hydrogen peroxide (H2O2)-induced lens epitheliums. Lens epithelium tissues were harvested from ARC or normal subjects (n = 23). CircSTRBP, spermatid perinuclear RNA binding protein (STRBP), and nicotinamide adenine dinucleotide phosphate oxidase subunit 4 (NOX4) levels were measured using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell proliferation, cycle progression, and apoptosis were assessed using 5-ethynyl-2'-deoxyuridine (EdU), Cell Counting Kit-8 (CCK-8), and flow cytometry assays. Caspase 3 activity, reactive oxygen species (ROS), malondialdehyde (MDA), and Glutathione peroxidases (GSH-PX) levels were detected using corresponding kits. NOX4 protein level was determined using Western blot. The interaction between insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) and circSTRBP or NOX4 was assessed through RNA immunoprecipitation (RIP). CircSTRBP and NOX4 abundances were increased in lens epithelium samples from ARC patients and H2O2-treated SRA01/04 cells. CircSTRBP knockdown might abolish H2O2-triggered SRA01/04 cell proliferation repression and apoptosis and oxidative stress promotion. In mechanism, circSTRBP is bound with IGF2BP1 and improves the stability and expression of NOX4 mRNA in SRA01/04 cells. CircSTRBP facilitated H2O2-induced SRA01/04 cell apoptosis and oxidative stress through by enhancing NOX4 mRNA stability via recruiting IGF2BP1, providing novel insights for ARC progression and treatment.

A novel rice grain size gene OsSNB was identified by genome-wide association study in natural population.

Increasing agricultural productivity is one of the most important goals of plant science research and imperative to meet the needs of a rapidly growing population. Rice (Oryza sativa L.) is one of the most important staple crops worldwide. Grain size is both a major determinant of grain yield in rice and a target trait for domestication and artificial breeding. Here, a genome-wide association study of grain length and grain width was performed using 996,722 SNP markers in 270 rice accessions. Five and four quantitative trait loci were identified for grain length and grain width, respectively. In particular, the novel grain size gene OsSNB was identified from qGW7, and further results showed that OsSNB negatively regulated grain size. Most notably, knockout mutant plants by CRISPR/Cas9 technology showed increased grain length, width, and weight, while overexpression of OsSNB yielded the opposite. Sequencing of this gene from the promoter to the 3'-untranslated region in 168 rice accessions from a wide geographic range identified eight haplotypes. Furthermore, Hap 3 has the highest grain width discovered in japonica subspecies. Compared to other haplotypes, Hap 3 has a 225 bp insertion in the promoter. Based on the difference between Hap 3 and other haplotypes, OsSNB_Indel2 was designed as a functional marker for the improvement of rice grain width. This could be directly used to assist selection toward an improvement of grain width. These findings suggest OsSNB as useful for further improvements in yield characteristics in most cultivars.

Clinical value of contrast-enhanced ultrasonography in focal hypoechogenic lesions of thyroid.

OBJECTIVE: The objectives of this study were to analyze the accuracy of contrast-enhanced ultrasonography (CE-US) in diagnosing focal hypoechogenic lesions of the thyroid (FHLT), and to explore the clinical value of CE-US in the diagnosis of FHLT. METHODS: Patients undergoing CE-US and ultrasound-guided fine needle aspiration (US-FNA) of FHLT at First Hospital of China Medical University between January 2017 and December 2018 were selected for the study; this included patients with papillary thyroid carcinoma (PTC), subacute thyroiditis (SAT) and focal Hashimoto thyroiditis (FHT). All patients underwent color Doppler ultrasonography (CD-US) after which thyroid image reporting and data system (TI-RADS) grading were done. Then, each patient underwent CE-US and US-FNA. The results of the CE-US were analyzed using descriptive statistics. The cytopathological results from the US-FNAs were the gold standard used to confirm the diagnoses. RESULTS: A total of 56 patients were selected for the study. In the PTC group (n = 16), grading was as follows: TI-RADS4a, n = 3; TI-RADS4b, n = 12; and TI-RADS4c, n = 1. More patients with PTC showed heterogeneous hypoenhancement (n = 15) than heterogeneous isoenhancement (n = 1) on CE-US. In the SAT group (n = 24), grading was as follows: TI-RADS3, n = 1; TI-RADS4a, n = 18; TI-RADS4b, n = 5. Fewer patients with SAT showed heterogeneous hypoenhancement (n = 2) than heterogeneous isoenhancement (n = 22) on CE-US. In the FHT group (n = 16), grading was as follows: TI-RADS3, n = 1; TI-RADS4a, n = 11; TI-RADS4b, n = 4. Of those in the FHT group, one patient showed heterogeneous isoenhancement, one patient showed heterogeneous hypoenhancement, and 14 showed uniform isoenhancement on CE-US. The diagnostic accuracy of CD-US alone differed significantly from that of CD-US + CE-US (p < 0.05). CONCLUSION: CE-US has a high diagnostic accuracy for FHLT and can be used to identify PTC, SAT, and FHT.

Non-invasive SERS serum detection technology combined with multivariate statistical algorithm for simultaneous screening of cervical cancer and breast cancer.

Surface-enhanced Raman scattering (SERS), as a rapid, reliable and non-destructive spectral detection technology, has made a series of breakthrough achievements in screening and pre-diagnosis of various cancerous tumors. In this paper, high-performance gold nanoparticles/785 porous silicon photonic crystals (Au NPs/785 PSi PhCs) active SERS substrates were specially designed for serum testing, and realized highly sensitive detection of serum from healthy people, patients with cervical cancer and breast cancer. Based on the SERS spectra of the three groups of serum, the significant differences between the healthy group and cancer group at 1030 cm-1 and 1051 cm-1 were analyzed, and the similar but different serum SERS spectra of cervical cancer and breast cancer patients were compared. In addition, the spectral difference detected by SERS technology combined with a multivariate statistical algorithm was used to distinguish three kinds of serum. The serum SERS spectral sensitive bands were extracted by recursive weighted partial least squares (rPLS), and the three classification diagnosis models were established by combining orthogonal partial least squares discriminant analysis (OPLS-DA), linear discriminant analysis (LDA) and principal component analysis support vector machine (PCA-SVM) for synchronous classification and discrimination of the three groups of serum. The diagnostic results showed that the overall screening accuracy of three models were 93.28%, 97.77% and 94.78%, respectively. These above results confirmed that the Au NPs/785 PSi PhCs can realize super-sensitive detection of serum, and the established diagnostic model has great potential for pre-diagnosis and simultaneous screening of cervical cancer and breast cancer.

An APETALA2/ethylene responsive factor, OsEBP89 knockout enhances adaptation to direct-seeding on wet land and tolerance to drought stress in rice.

Water stress is the most important adverse factor limiting rice production. Too much water leads to flood and too little leads to drought. Floods and droughts can severely damage crop at different times of the rice life cycle. So the research on submergence tolerance and drought resistance of rice is particularly urgent. In this study, we reported that OsEBP89 (Oryza sativa Ethylene-responsive element binding protein, clone 89), a member of the AP2/ERF subfamily, is involved in a novel signal transduction associated with the tolerance to drought and submergence stress. OsEBP89 was found to be strongly inhibited by drought stress and promoted by submergence. The OsEBP89 protein was located at the nucleus in the rice protoplast. Loss of OsEBP89 was found to improve the seed germination under submerged conditions and also enhanced the tolerance to drought stress throughout growth stage. Additionally, OsEBP89 knockout rice plants increased the accumulation of proline, improved the ability to scavenge ROS compared to overexpression lines and wild type after PEG treatment. Transcriptome data indicates that knockout of OsEBP89 improved the expression of specific genes in response to adverse factors, such as OsAPX1, OsHsfA3, and OsP5CS. Further results indicate that OsEBP89 can interact with and be phosphorylated by SnRK1α (sucrose non-fermenting-1-related protein kinase-1 gene). These findings provide insight into the mechanism of abiotic stress tolerance, and suggest OsEBP89 as a new genetic engineering resource to improve abiotic stress tolerance in rice.

Microwave method synthesis of magnetic ionic liquid/gold nanoparticles as ultrasensitive SERS substrates for trace clopidol detection.

Clopidol is one of the most widely used anti-coccidiosis drugs. Its residues in poultry products and the environment pose a serious threat to human health. In this work, microwave-assisted synthesis of magnetic ionic liquid/gold nanoparticles (MIL-Au NPs) as the SERS substrates were first designed for sensitive and reliable determination of clopidol residue in egg samples. The experiment shows that MIL(1-methyl-3-hexyl imidazole ferric tetrachloride ([C6mim]FeCl4)) and microwave play a key role in the dispersion and morphology of Au NPs. Under the optimal conditions, the as-prepared MIL-Au NPs were applied to the SERS detection of clopidol in methanol and egg solution and its detection limits can be as low as to 0.5 µg/kg (equal to 0.5 ppb) in both solutions. The standard curves with regression coefficients of 0.9298 and 0.93496 were constructed in the linear range of 100-1000 ppb and 0.5-50 ppb for clopidol in egg solutions. Moreover, satisfactory recoveries (97.5-103.2%) were obtained for egg samples. The developed SERS method provides a way for quantitation of clopidol and can be applied for the convenient, reliable, and highly sensitive detection of antibiotic residues in food and environment, which has great potential in food safety and biological monitoring. Graphical abstract.

Gastric cancer vaccines synthesized using a TLR7 agonist and their synergistic antitumor effects with 5-fluorouracil.

BACKGROUND: Vaccines play increasingly important roles in cancer treatment due to their advantages of effective targeting and few side effects. Our laboratory has attempted to construct vaccines by conjugating TLR7 agonists with tumor-associated antigens. Furthermore, immunochemotherapy has recently become an appealing approach to cancer therapy. 5-fluorouracil (5-FU), a commonly used chemotherapeutic agent, can reportedly potently and selectively kill tumor-associated MDSCs in vivo. METHODS: Gastric cancer vaccines were synthesized by the covalent attachment of our TLR7 agonist with the gastric cancer antigen MG7-Ag tetra-epitope, leading to T7 - ML (linear tetra-epitope) and T7 - MB (branched tetra-epitope). Cytokines induced by the vaccines in vitro were assessed by ELISA. A tumor challenge model was created by treating BALB/c mice on either a prophylactic or therapeutic vaccination schedule. 5-FU was simultaneously applied to mice in the combination treatment group. CTL and ADCC activities were determined by the LDH method, while CD3+/CD8+, CD3+/CD4+ T cells and MDSCs were evaluated by flow cytometry. RESULTS: In vitro, rapid TNF-α and IL-12 inductions occurred in BMDCs treated with the vaccines. In vivo, among all the vaccines tested, T7 - MB most effectively reduced EAC tumor burdens and induced CTLs, antibodies and ADCC activity in BALB/c mice. Immunization with T7 - MB in combination with 5-FU chemotherapy reduced tumor sizes and extended long-term survival rates, mainly by improving T cell responses, including CTLs, CD3+/CD8+ and CD3+/CD4+ T cells. 5-FU also enhanced the T7 - MB efficiency by reversing immunosuppressive factors, i.e., MDSCs, which could not be validly inhibited by the vaccines alone. In addition, T7 - MB repressed tumor growth and immune tolerance when the therapeutic schedule was used, although the effects were weaker than those achieved with either T7 - MB alone or in combination with 5-FU on the prophylactic schedule. CONCLUSIONS: A novel effective gastric cancer vaccine was constructed, and the importance of branched multiple antigen peptides and chemical conjugation to vaccine design were confirmed. The synergistic effects and mechanisms of T7 - MB and 5-FU were also established, observing mainly T cell activation and MDSC inhibition.

TRPA1 contributed to the neuropathic pain induced by docetaxel treatment.

Peripheral mechanical neuropathic pain is a serious side effect of docetaxel chemotherapy for cancer. However, the underlying mechanism for this side effect is unknown. In the present study, we found that docetaxel treatment induced mechanical allodynia in rats. We further revealed that the transient receptor potential ankyrin subtype 1 protein (TRPA1) protein level is upregulated and the TRPA1 activator allyl isothiocyanate induced larger ion currents in the dorsal root ganglion neurons from the docetaxel treated rats. In addition, application the TRPA1 blocker Ap18 reversed the docetaxel-induced mechanical hypersensitivity. We suggest that the docetaxel-induced mechanical allodynia is mediated by upregulation of TRPA1 in dorsal root ganglion neurons.

Androgenesis, gynogenesis, and parthenogenesis haploids in cucurbit species.

Haploids and doubled haploids are critical components of plant breeding. This review is focused on studies on haploids and double haploids inducted in cucurbits through in vitro pollination with irradiated pollen, unfertilized ovule/ovary culture, and anther/microspore culture during the last 30 years, as well as comprehensive analysis of the main factors of each process and comparison between chromosome doubling and ploidy identification methods, with special focus on the application of double haploids in plant breeding and genetics. This review identifies existing problems affecting the efficiency of androgenesis, gynogenesis, and parthenogenesis in cucurbit species. Donor plant genotypes and surrounding environments, developmental stages of explants, culture media, stress factors, and chromosome doubling and ploidy identification are compared at length and discussed as methodologies and protocols for androgenesis, gynogenesis, and parthenogenesis in haploid and double haploid production technologies.

Perturbation of physiological systems by nanoparticles.

Nanotechnology is having a tremendous impact on our society. However, societal concerns about human safety under nanoparticle exposure may derail the broad application of this promising technology. Nanoparticles may enter the human body via various routes, including respiratory pathways, the digestive tract, skin contact, intravenous injection, and implantation. After absorption, nanoparticles are carried to distal organs by the bloodstream and the lymphatic system. During this process, they interact with biological molecules and perturb physiological systems. Although some ingested or absorbed nanoparticles are eliminated, others remain in the body for a long time. The human body is composed of multiple systems that work together to maintain physiological homeostasis. The unexpected invasion of these systems by nanoparticles disturbs normal cell signaling, impairs cell and organ functions, and may even cause pathological disorders. This review examines the comprehensive health risks of exposure to nanoparticles by discussing how nanoparticles perturb various physiological systems as revealed by animal studies. The potential toxicity of nanoparticles to each physiological system and the implications of disrupting the balance among systems are emphasized.

Nestin positively regulates the Wnt/ß-catenin pathway and the proliferation, survival and invasiveness of breast cancer stem cells.

INTRODUCTION: We investigated Nestin expression in triple-negative breast cancer and examined how the modulation of Nestin expression affects cell cycle progression, survival, invasion and regulatory signaling in breast cancer stem cells (CSC) in vitro. METHODS: Nestin expression in 150 triple-negative breast cancer specimens were examined by immunohistochemistry. The role of Nestin expression in tumorigenesis was examined by assaying naturally occurring Nestinhigh/Nestinlow CSC from 12 breast cancer tissues, as well as CSC from 26 clinical specimens, where Nestin overexpression and silencing was achieved by genetic manipulation, for their ability to form mammospheres and induce solid tumors. Cell cycle progression, spontaneous apoptosis and invasiveness of Nestin-silenced breast CSC were investigated by flow cytometry and transwell assays. The relative levels of expression of epithelial-mesenchymal transition (EMT) and Wnt/ß-catenin pathway-related molecules were determined by western blotting. RESULTS: Nestin expression was significantly associated with poor survival in patients with triple-negative breast cancer (P = 0.01). Nestinhigh breast CSC rapidly formed typical mammospheres in vitro. Nestinhigh, but not Nestinlow CSC, efficiently formed solid tumors in vivo. Nestin silencing induced cell cycle arrest at G2/M (52.03% versus 19.99% in controls) and promoted apoptosis (36.45% versus 8.29% in controls). Nestin silencing also inhibited breast CSC invasiveness, and was associated with significantly upregulated E-cadherin, while N-cadherin, vimentin, a-smooth muscle actin (a-SMA), matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) expression was downregulated (P <0.05 for all). Nestin silencing also upregulated Axin, glycogen synthase kinase-3 beta (GSK-3ß), adenomatous polyposis coli (APC), and peroxisome proliferator-activated receptor alpha (PPARa), and downregulated ß-catenin, c-Myc, cyclin D and MMP-7 expression in CSC. Inhibition of the Wnt/ß-catenin pathway mitigated mammosphere formation in Nestinhigh CSC, while inhibition of GSK-3ß promoted the mammosphere formation in Nestinlow CSC (P <0.05 for all). CONCLUSIONS: Our data indicates that Nestin positively regulates the proliferation, survival and invasiveness of breast CSC by enhancing Wnt/ß-catenin activation.

Clinical implications of GRHL3 protein expression in breast cancer.

The objective of this study was to investigate the expression and significance of the GRHL3 protein in breast cancer diagnosis and prognosis. Overall, 111 patients with histologically confirmed breast cancer who had undergone radical surgery were enrolled in this study. The GRHL3 protein expression status in the breast cancers was evaluated by immunohistochemistry. The relationship between GRHL3 protein expression status and clinicopathological factors and the breast cancer prognoses was also determined. In total, 71 (63.96%) out of 111 cases were found to express GRHL3 protein. GRHL3 expression was higher in breast cancers, compared to other pathologic cancer types (χ (2) = 5.68, P < 0.05). Moreover, GRHL3 protein was also observed to correlate with breast cancer clinical stage and histological grade (χ (2) = 7.99, P < 0.05 and χ (2) = 7.907, P < 0.05, respectively). Interestingly, triple-negative breast cancers had lower expression rates than other breast cancers (41.18 vs. 71.28%, P < 0.05). GRHL3 was shown to be an independent prognostic factor of breast cancer in Cox regression analysis. Altogether, our results indicate predominant GRHL3 expression in breast cancers, especially non-triple-negative cancers and early stage cancers. GRHL3 expression appeared to decrease with tumor progression. Survival analysis demonstrated the inhibitory effect of GRHL3 in breast cancer. These results strongly suggest the possible involvement of GRHL3 in tumor suppression.

Nestin: predicting specific survival factors for breast cancer.

The purpose of this study is to investigate the relationship between nestin expression and clinicopathological characteristics, immunohistochemical markers and to determine the prognostic impact of nestin expression in breast cancer so as to lay a foundation for the treatment of breast cancer. A total of 109 patients who were histologically diagnosed with breast cancer and underwent radical operations from January 2006 to September 2007 in China Medical University were enrolled in the study. Nestin protein expression was evaluated by immunohistochemistry. The relationship between nestin and other parameters was analyzed by using chi-square test and Fisher's exact test. Nestin expression was observed in 37.6% (41/109) of cases. There were no significant differences between the age of >40 and ≤40 years group in terms of nestin expression (39.8 vs 18.2%; P = 0.161). The rate of nestin expression between those with and without lymph node metastasis was not significantly different (X (2) = 0.086; P = 0.769). The 5-year survival rates of the patients with nestin expression and those without were 34.1% (14/41) and 55.9% (38/68), respectively (P = 0.028). Overall, triple-negative breast cancers had higher expression rates than other cancers (54.1 vs 29.2%; P = 0.011). Nestin expression rate in ER- and PR-negative tumors was found to be significantly higher than cases that were ER- and PR-positive (P = 0.011 and P = 0.036, respectively). However, it was not found that HER2 expression was related to nestin expression (P = 0.120). These results suggest that the expression of nestin might play an important role in the prognosis of breast carcinoma, especially in the triple-negative subgroups.

Synthesis and immunoregulatory activities of conjugates of a Toll-like receptor 7 inert ligand.

In the synthesis and modification of the analogs of an adenine type of Toll-like receptor (TLR) 7 agonists, we found a special compound, 9-propionyloxy-8-hydroxy-2-(2-methoxyethoxy)-adenine (6). It is a synthesized TLR7 inert ligand, which does not respond to TLR7 itself. However, it can be coupled with protein or peptide antigens via propionyloxy functional group to promote their immunogenicity significantly. The compound was covalently coupled to protein and peptide to get the conjugates. The inductivity of cytokine production by the conjugates was 872.4-fold compared with the unconjugated antigens in vitro by mouse splenocyte. These data show that the immunostimulatory activity of inert TLR7 ligand can be endowed, and the activity of antigens can be amplified by conjugation with various proteins and peptides, thus broadening the potential therapeutic application and reducing the risk of TLR7 agonists' side effects.

In-Situ Fabrication of Electroactive Cu2+-Trithiocyanate Complex and Its Application for Label-Free Electrochemical Aptasensing of Thrombin.

The preparation of an electroactive matrix for the immobilization of the bioprobe shows great promise to construct the label-free biosensors. Herein, the electroactive metal-organic coordination polymer has been in-situ prepared by pre-assembly of a layer of trithiocynate (TCY) on a gold electrode (AuE) through Au-S bond, followed by repetitive soaking in Cu(NO3)2 solution and TCY solutions. Then the gold nanoparticles (AuNPs) and the thiolated thrombin aptamers were successively assembled on the electrode surface, and thus the electrochemical electroactive aptasensing layer for thrombin was achieved. The preparation process of the biosensor was characterized by an atomic force microscope (AFM), attenuated total reflection-Fourier transform infrared (ATR-FTIR), and electrochemical methods. Electrochemical sensing assays showed that the formation of the aptamer-thrombin complex changed the microenvironment and the electro-conductivity of the electrode interface, causing the electrochemical signal suppression of the TCY-Cu2+ polymer. Additionally, the target thrombin can be label-free analyzed. Under optimal conditions, the aptasensor can detect thrombin in the concentration range from 1.0 fM to 1.0 µM, with a detection limit of 0.26 fM. The spiked recovery assay showed that the recovery of the thrombin in human serum samples was 97.2-103%, showing that the biosensor is feasible for biomolecule analysis in a complex sample.

Characterization of a chlorine resistant and hydrophilic TiO2/calcium alginate hydrogel filtration membrane used for protein purification maintaining protein structure.

The development of membranes for protein purification has stringent requirement of disinfection resistance, low protein adsorption and anti-fouling, without changing protein structure. In this study, hydrophilic titanium dioxide (TiO2)/calcium alginate (TiO2/CaAlg) hydrogel membranes were prepared by a simple ionic cross-linking method. The effects of the porogenic agent polyethylene glycol (PEG) concentration, the molecular weight of PEG, and the concentration of TiO2 on the filtration properties were systematically investigated. The TiO2/CaAlg membrane exhibited excellent bovine serum albumin (BSA) rejection and anti-fouling properties. The mechanical properties and surface energy of the TiO2/CaAlg membrane were significantly improved. The chemical bonding mechanism of TiO2 and NaAlg was investigated by molecular dynamic simulation. The TiO2/CaAlg membrane had good chlorine resistance and could be disinfected or cleaned with sodium hypochlorite. The TiO2/CaAlg hydrogel membrane loaded with polyhydroxybutyrate (PHB) nanofibers maintained high flux (136.7 L/m2h) and high BSA rejection (98.0 %) at 0.1 MPa. The results of circular dichroism and synchronous fluorescence indicated that the secondary structure of BSA was maintained after membrane separation. This study provides one method for the preparation of green and environmentally friendly membrane for protein purification.

Self-assembled cationic micelles based on PEG-PLL-PLLeu hybrid polypeptides as highly effective gene vectors.

Developing safe and effective nonviral gene vector is highly crucial for successful gene therapy. In the present study, we designed a series of biodegradable micelles based on hybrid polypeptide copolymers of poly(ethylene glycol)-b-poly(l-lysine)-b-poly(l-leucine) (PEG-PLL-PLLeu) for efficient gene delivery. A group of amphiphilic PEG-PLL-PLLeu hybrid polypeptide copolymers were synthesized by ring-opening polymerization of N-carboxyanhydride, and the chemical structure of each copolymer was characterized by (1)H NMR and FT-IR spectroscopy measurement. The PEG-PLL-PLLeu micelles were positively charged with tunable sizes ranging from 40 to 90 nm depending on the length of PLL and PLLeu segment. Compared with PEG-PLL copolymers, PEG-PLL-PLLeu micelles demonstrated significantly higher transfection efficiency and less cytotoxicity. Furthermore, the transfection efficiency and biocompatibility of the micelles can be simultaneously improved by tuning the length of PLL and PLLeu segments. The transfection efficiency of PEG-PLL-PLLeu micelles in vivo was two to three times higher than that of PEI(25k), which was attributable to their capability of promoting DNA condensation and cell internalization as well as successful lysosome escape. Hence well-defined PEG-PLL-PLLeu micelles would serve as highly effective nonviral vectors for in vivo gene delivery.

Establishment of a reliable scheme for obtaining highly stable SERS signal of biological serum.

As a rapid and non-destructive biological serum detection method, SERS technology was widely used in the screening and medical diagnosis of various diseases by combining the analysis of serum SERS spectrum and multivariate statistical algorithm. Because of the high complexity of serum components and the variability of SERS spectra, which often resulted in the phenomenon that the SERS spectrum of the same biological serum was significantly different due to the different test conditions. In this experiment, through the dilution treatment of the serum and the systematic test of the serum of all concentration gradients with lasers of wavelength of 785, 633 and 532 nm, the most suitable conditions for detecting the serum were investigated. The experimental results showed that only when the serum is diluted to low concentration (10 ppm), the SERS spectrum with high reproducibility and stability could be obtained, furthermore, the low concentration serum had weak tolerance to laser, and 532 nm laser was not suitable for serum detection. In this paper, a set of test scheme for obtaining highly stable serum SERS spectra was established by using high-performance gold nanoparticles (Au NPs) as the active substrate of SERS. Through comparative analysis of SERS spectrum of serum of normal people and cervical cancer, the reliability of the established low-concentration serum test program was verified, as well as its great potential advantages in disease screening and diagnosis.

Associations of BRAF V600E, clinical pathology and imaging factors with the recurrence rate of papillary thyroid microcarcinoma.

In the present study, the recurrence rate of papillary thyroid microcarcinoma (PTMC) was assessed by analyzing postoperative follow-up data of affected patients and its associations with BRAF V600E, clinical pathology and imaging factors were explored. A total of 506 patients with PTMC were selected who underwent surgery from January 2014 to March 2016. The maximal diameter of thyroid nodules was ≤1 cm and all patients who underwent BRAF V600E testing and evaluation for lymph node metastasis. Postoperatively, each patient was regularly followed up to detect recurrence. Categorical variables were comparatively analyzed using univariate Cox linear regression analysis to screen for protective and adverse factors influencing recurrence of PTMC. A stepwise Cox proportional hazards regression model analysis was performed to explore risk factors affecting recurrence. Among the 506 patients, 477 were followed up, 29 were lost to follow-up and 26 patients experienced recurrence. The 5-year recurrent rate of PTMC was 5.45%. The univariate Cox regression analysis indicated that PTMC recurrence was influenced by BRAF V600E, sex, multifocality, capsular invasion and lateral cervical lymph node metastasis (P<0.05), but not by age, tumor location on the thyroid, size, single central lymph node metastasis, distant metastasis and operative approach (P>0.05). The significant factors associated with recurrent PTMC were subjected to stepwise multivariate Cox proportional hazards regression model analysis and the results indicated that BRAF V600E, sex, multifocality and lateral cervical lymph node metastasis were independent factors influencing recurrence in patients with PTMC, with a statistically significant difference (P<0.05). In conclusion, BRAF V600E, sex, multifocality and lateral cervical lymph node metastasis are independent risk factors for recurrent PTMC.

Correlation between FAK and EGF-Induced EMT in Colorectal Cancer Cells.

Epithelial-mesenchymal transition (EMT) plays an important role in the invasion and metastasis of colorectal cancer, which is mediated by FAK and EGF. However, whether FAK participates in EMT in colorectal cancer cells through the EGF/EGFR signaling pathway remains unknown. The aim of this study was to investigate the effector mechanisms of FAK in the process of EGF-induced EMT in colorectal cancer cells and to determine whether miR-217 is involved in this process. Caco-2 cancer cells were routinely cultured with and without treatment with 100 ng/mL EGF, and changes in cell morphology were observed using an inverted microscope. In addition, a transwell assay was used to detect cell migration under the condition of EGF treatment. The expression of FAK, pFAK, E-cadherin, vimentin, and ß actin was assessed by western blotting, and the expression of miR-217 was assessed using real-time PCR. We found that EGF induced EMT in colorectal cancer cells and enhanced cell migration and invasion ability. Moreover, FAK was involved in the EGF-induced EMT of colorectal cancer cells. EGF upregulated the expression of E-cadherin in colorectal cancer cells by activating FAK, and miR-217 was found to participate in EGF-induced EMT in colorectal cancer cells. Our findings indicate that EGF induces EMT in colorectal cancer cells by activating FAK, and miR-217 is involved in the EGF/FAK/E-cadherin signaling pathway.