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Context: The timing of early cumulus cell removal (ECCR) can be changed within a range. The change has an effect on the multiple pronuclei (MPN) rate and the exposure time of oocytes to sperm waste products. The timing of ECCR effects the outcomes of assisted reproductive technology, however, it is still unclear what time is best for ECCR. Aims: To find the best time for ECCR based on clinical outcomes in order to increase the success rate of assisted reproductive technology. Methods: A retrospective study was performed. Cycles were categorised into six groups according to the timing of ECCR. The clinical outcomes of these six groups were compared by Kruskal–Wallis test and Pearson X 2 test. Key results: The timing of ECCR had a significant effect on the MPN rate, 0PN without cleavage rate and grade 1–2 embryo rate at Day 3. Among our six time groups of ECCR, the cumulus cell removal ≤4 h post-insemination group had the highest MPN rate and grade 1–2 embryo rate at Day 3, and the 5.5 h < time ≤6 h group had the highest pronuclear formation failure rate. In addition, 2PN rate was not significantly different among the six time groups. Conclusions: Within the time range of ECCR in our study, selecting an earlier timing to remove the cumulus cells may have a better outcome. Implications: ECCR is helpful for optimisation of the short-term insemination strategy. An important operation of assisted reproductive technology is cumulus cell removal. The timing of cumulus cell removal has an effect on the outcomes of assisted reproductive technology. Our study aims to find the best time for cumulus cell removal, which will increase the success rate of assisted reproductive technology. [ABSTRACT FROM AUTHOR]

Similar to other immune-mediated diseases, IgG4-related disease (IgG4-RD) is the disease that develops in genetically susceptible individuals exposed to external or endogenous antigens. In the present study, it was confirmed that MAG (myelin-associated glycoprotein) antibodies (IgG, IgG4, and IgM) were detected by immunofluorescence (IFA) in serum of the patients with IgG4-RD. In vivo, the levels of prolactin and Th2 cytokines in CGRP+/− rats were higher than those in wild-type. Our findings indicate that the presence of CGRP-deficiency-mediated MAG antibodies is a probable molecular basis for the initial events which were triggered in IgG4-RD immune responses via prolactin upregulation. [ABSTRACT FROM AUTHOR]

• Adropin activates PPAR-γ and its downstream target gene CD36 can take up exogenous fatty acids. • Adropin can mobilize macrophages to break down store lipids, take in exogenous fatty acids and promote lipid metabolism. • Adropin regulates the repolarization of macrophages from M1 to M2, reverses the inflammatory phenotype, and exerts anti-inflammatory effects. • The level of serum adropin in IBD patients was decreased and negatively correlated with the severity of IBD. The aim of this study was to describe the effects of adropin deficiency on the distribution, phenotype and pathological phenotype of macrophages in colonic and mesenteric tissues of AdrKO (Enho −/−) mice, so as to explore the mechanism of adropin deficiency in spontaneous and experimental colitis. In this study, RNA-seq and metabonomics were used to screen the regulatory mechanism of adropin on the phenotypic transformation of macrophages. We found that adropin levels in active UC patients were significantly lower than those in normal subjects and remission UC patients, and at the same time, a large number of proinflammatory M1-type macrophages were infiltrated in the mesenteric tissue of colonic tissues from UC and CD patients. At the same time, spontaneous colitis occurred in Enho-/- (adropin-deficient)C57BL/6 mice, and there was an imbalance of M2 → M1 polarization of macrophages in colon and mesentery of Enho-/- mice. In vivo , it has showed that adropin deficiency could exacerbate the pathological phenotype of colitis induced by TNBS. In vitro, adropin was used to intervene RAW264.7 macrophages, and then combined analysis of RNA-seq and metabolomics demonstrated that adropin regulated lipid metabolism of macrophages through PPARγ, thus promoting the repolarization of macrophages from M1 to M2. Adropin deficiency led to an imbalance in the phenotypic distribution of macrophages infiltrating the colon and mesenteric tissues, namely, an increase in M1 type, which led to the occurrence and development of colitis. [ABSTRACT FROM AUTHOR]

Herein, polythionine-Au nanocomposites (PTh-Au) were prepared through one-pot reaction using HAuCl 4 as co-oxidant in thionine solution. The prepared PTh-Au exhibited intrinsic electrochemical activity based on the dual participation of protons and electrons during electrochemical doping/dedoping process. Thus, a label-free electrochemical immunoassay to detect a high isoelectric point (p I ) biomarker (using trypsin as a model) was developed based on co-participation of proton and electron in the PTh-Au's response. In a detecting buffer with pH values lower than trypsin's p I (=10.8), positively charged trypsin prevented proton and electron transfer simultaneously to reduce the current response of PTh-Au. The decreasing amplitude of the current can be used to quantify the concentration of trypsin, which can be utilized to determine the health condition of pancreas. The proposed immunoassay exhibited wide linear range, low limit of detection and high specificity in a detecting buffer (pH 5.5). The obtained immunosensor was successfully applied to detect trypsin in human serum accurately. Additionally, this sensing strategy may be used to detect other important biomarkers with relatively high p I s (>7). [ABSTRACT FROM AUTHOR]

Actinic keratosis (AK) is a common precancerous skin lesion with significant harm, and it is often confused with non-actinic keratoses (NAK). At present, the diagnosis of AK mainly depends on clinical experience and histopathology. Due to the high difficulty of diagnosis and easy confusion with other diseases, this article aims to develop a convolutional neural network that can efficiently, accurately, and automatically diagnose AK. This article improves the MobileNet model and uses the AK and NAK images in the HAM10000 dataset for training and testing after data preprocessing, and we performed external independent testing using a separate dataset to validate our preprocessing approach and to demonstrate the performance and generalization capability of our model. It further compares common deep learning models in the field of skin diseases (including the original MobileNet, ResNet, GoogleNet, EfficientNet, and Xception). The results show that the improved MobileNet has achieved 0.9265 in accuracy and 0.97 in Area Under the ROC Curve (AUC), which is the best among the comparison models. At the same time, it has the shortest training time, and the total time of five-fold cross-validation on local devices only takes 821.7 s. Local experiments show that the method proposed in this article has high accuracy and stability in diagnosing AK. Our method will help doctors diagnose AK more efficiently and accurately, allowing patients to receive timely diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE) leads to dysfunctions of human trophoblast cells and further induces miscarriage. In our previous study, we have found that lnc-HZ03 and miR-hz03 are upregulated in BPDE-exposed human trophoblast cells and in recurrent miscarriage tissues; and the upregulated miR-hz03 caused by lnc-HZ03 further promotes the apoptosis of human trophoblast cells and induces miscarriage. However, how lnc-HZ03 upregulates miR-hz03 is completely unknown. In this study, we find that lnc-HZ03 upregulates the expression level of a transcription factor TFIID (a TATA-binding protein) and promotes TFIID-mediated transactivation response element RNA-binding protein (TRBP) transcription. Subsequently, the upregulated TRBP promotes the maturation of miR-hz03 by splicing its precursor pre-miR-hz03 in human trophoblast cells. In BPDE-exposed trophoblast cells or in recurrent miscarriage tissues, lnc-HZ03 was upregulated, which accelerates the TFIID-mediated TRBP transcription and thus promotes TRBP-mediated miR-HZ03 maturation. Subsequently, the upregulated miR-hz03 further promotes the apoptosis of human trophoblast cells and induces miscarriage. This work provides new insights into the regulation of miRNA expression levels by lncRNAs in BPDE-exposed human trophoblast cells. [ABSTRACT FROM AUTHOR]

We used a releasable disulfide carbonate linker to construct precursors of gelators and form stable gels. [ABSTRACT FROM AUTHOR]

Background and Aims: The high mortality rate of pancreatic cancer is a bottleneck for further treatment with long-term efficacy. Thus, it is urgent to identify new methods to accurately predict the early onset of pancreatic cancer. We hypothesized that the different genotypes of cationic trypsinogen ( PRSS1) gene could confer susceptibility and/or resistance to pancreatic cancer in the Han Chinese population. Methods: The genotypes of PRSS1 were determined in 154 patients with pancreatic cancer and in a control group of 520 healthy individuals of Han Chinese descent. Clinical information was obtained, single-nucleotide polymorphisms (SNPs) of the PRSS1 gene were analyzed by direct sequencing, and the distribution of the genotypes were tested for Hardy-Weinberg equilibrium. Odds ratios and 95% confidence intervals were calculated by logistic regression analysis to estimate the associations between the different genotypes or haplotypes and the risk of pancreatic cancer. Results: Three SNPs (−409 C/T, −204 A/C, and c.486 C/T) were identified. A case-control analysis revealed a 0.118-fold (95% CI: 0.037-0.653), 0.842-fold (95% CI: 0.177-4.010), and 0.750-fold (95% CI: 0.519-1.085) change in risk of developing pancreatic cancer for individuals harboring these SNPs, respectively. The individuals with the −409 C/T genotype tended to have a reduced risk compared to those who carried the −409 T/T genotype. A protective effect was observed for the C-A-C haplotype compared to the T-A-T haplotype (OR = 0.115, 95% CI: 0.016-0.849) or compared to the T-A-C haplotype (OR = 0.090, 95% CI: 0.012-0.667). Serum levels of trypsin in patients with the −409 C/T genotype were only one-fourth that of those with the −409 T/T genotype and only one-third that of the healthy controls. Conclusions: The −409 C/T genotype of PRSS1 was reveled to be a protective factor against pancreatic cancer in the Han Chinese population. [ABSTRACT FROM AUTHOR]

Abstract: In this study, an enzyme-amplified electrochemical biosensor was developed for detection of the promyelocytic leukemia/retinoic acid receptor alpha (PML/RARα) fusion gene in acute promyelocytic leukemia (APL). This new sensor employs a hairpin locked nucleic acids (LNAs) probe dually labeled with biotin and carboxyfluorescein molecule (FAM). The probe is immobilized at a streptavidin-modified electrode surface via the biotin–streptavidin bridge, and FAM serves as an affinity tag for the peroxidase conjugate binding. Initially, the immobilized hairpin probe was in the “closed” state in the absence of the target, which shielded FAM from being approached by the bulky anti-FAM-HRP conjugate due to the steric effect. Target binding opens the hairpin structure of the probe, the probe undergoes a significant conformational change, forcing FAM away from the electrode. As a result, the FAM label becomes accessible by the anti-FAM-HRP, and the target hybridization event can be sensitively transduced via the enzymatically amplified electrochemical current signal. This new biosensor demonstrates its excellent specificity for single-base mismatch and able to detect as little as 83fM target DNA even in the presence of human serum. We also employed this sensor to directly detect PCR real sample with satisfactory results. [Copyright &y& Elsevier]

Background: Understanding factors associated with antiretroviral treatment (ART) adherence is crucial for ART success among people living with HIV (PLHIV) in the "test and treat" era. Multiple psychosocial factors tend to coexist and have a syndemic effect on ART adherence. We aimed to explore factors associated with ART adherence and the syndemic effect of multiple psychosocial factors on ART adherence among PLHIV newly starting ART in Guangdong Province, China. Methods: Newly diagnosed PLHIV from six cities in Guangdong Province were recruited between May 2018 and June 2019, and then followed up from May 2019 to August 2020. Baseline and follow-up data were collected from a questionnaire and the national HIV surveillance system, the follow-up data of which were analyzed in this study. A Center for Adherence Support Evaluation (CASE) index > 10 points was defined as optimal ART adherence, which was measured via participants' self-reported adherence during follow-up survey. Multivariable logistic regression was used to identify factors associated with ART adherence. Exploratory factor analysis (EFA) and multi-order latent variable structural equation modeling (SEM) were performed to explore the syndemic effect of multiple psychosocial factors on ART adherence. Results: A total of 734 (68.53%) follow-up participants were finally included in this study among the 1071 baseline participants, of whom 91.28% (670/734) had self-reported optimal ART adherence. Unemployment (aOR = 1.75, 95%CI: 1.01–3.02), no medication reminder (aOR = 2.28, 95%CI: 1.09–4.74), low medication self-efficacy (aOR = 2.28, 95%CI: 1.27–4.10), low social cohesion (aOR = 1.82, 95%CI: 1.03–3.19), no social participation (aOR = 5.65, 95%CI: 1.71–18.63), and ART side effects (aOR = 0.46, 95%CI: 0.26–0.81) were barriers to optimal ART adherence. The EFA and second-order latent variable SEM showed a linear relationship (standardized coefficient = 0.43, P < 0.001) between ART adherence and the latent psychosocial (syndemic) factor, which consisted of the three latent factors of medication beliefs and self-efficacy (standardized coefficient = 0.65, P < 0.001), supportive environment (standardized coefficient = 0.50, P < 0.001), and negative emotions (standardized coefficient=-0.38, P < 0.01). The latent factors of medication beliefs and self-efficacy, supportive environment, and negative emotions explained 42.3%, 25.3%, and 14.1% of the variance in the latent psychosocial factor, respectively. Conclusions: About nine out of ten PLHIV on ART in Guangdong Province self-reported optimal ART adherence. However, more efforts should be made to address barriers to optimal ART adherence. [ABSTRACT FROM AUTHOR]

Background: Previous studies revealed somatic mutations of the cationic trypsinogen gene (PRSS1) in patients with chronic pancreatitis and pancreatic cancer. However, whether PRSS1 mutations trigger pancreatic cancer and/or promote malignant proliferation and metastasis in pancreatic cancer remains largely unclear, as well as the potential underlying mechanisms. Methods: In the present study, whole-exome sequencing was applied for screening, and the R116C mutation was validated by Sanger sequencing. Phosphorylation antibody array, RNA-Seq, and RT-qPCR were adopted to screen and validate that R116C mutation promoted pancreatic cancer progression via the JAK1-STAT5 pathway. Results: It showed that migration and invasion were significantly increased in R116C-bearing PANC-1 cells compared with wild type counterparts. In a transgenic mouse model of iZEG-PRSS1_R116C, primary pancreatic intraepithelial neoplasia (PanINs) was observed in the pancreatic duct. Conclusions: These findings suggested a novel pathway mediating pancreatic cancer development, with PRSS1 mutation and overexpression playing an "inside job" role in pancreatic carcinogenesis and tumor development. [ABSTRACT FROM AUTHOR]

In response to the pressing need for robust disease diagnosis from gastrointestinal tract (GIT) endoscopic images, we proposed FLATer, a fast, lightweight, and highly accurate transformer-based model. FLATer consists of a residual block, a vision transformer module, and a spatial attention block, which concurrently focuses on local features and global attention. It can leverage the capabilities of both convolutional neural networks (CNNs) and vision transformers (ViT). We decomposed the classification of endoscopic images into two subtasks: a binary classification to discern between normal and pathological images and a further multi-class classification to categorize images into specific diseases, namely ulcerative colitis, polyps, and esophagitis. FLATer has exhibited exceptional prowess in these tasks, achieving 96.4% accuracy in binary classification and 99.7% accuracy in ternary classification, surpassing most existing models. Notably, FLATer could maintain impressive performance when trained from scratch, underscoring its robustness. In addition to the high precision, FLATer boasted remarkable efficiency, reaching a notable throughput of 16.4k images per second, which positions FLATer as a compelling candidate for rapid disease identification in clinical practice. [ABSTRACT FROM AUTHOR]

The article discusses a study which examined factors associated with immediate antiretroviral therapy (ART) initiation among newly diagnosed people living with HIV (PLHIV) aged 18 years and above between May 2018 and June 2019 at six study sites in Guangdong Province, China. Topics include characteristics of the study participants, factors associated with immediate ART initiation among newly diagnosed PLHIV in Guangdong Province, and limitations of the study.

Objective: Welding fume exposure is inevitable of welding workers and poses a severe hazard to their health since welding is a necessary industrial process. Thus, preclinical diagnostic symptoms of worker exposure are of great importance. The aim of this study was to screen serum differential metabolites of welding fume exposure based on UPLC–QTOF-MS/MS. Methods: In 2019, 49 participants were recruited at a machinery manufacturing factory. The non-target metabolomics technique was used to clarify serum metabolic signatures in people exposed to welding fume. Differential metabolites were screened by OPLS-DA analysis and Student's t-test. The receiver operating characteristic curve evaluated the discriminatory power of differential metabolites. And the correlations between differential metabolites and metal concentrations in urine and whole blood were analyzed utilizing Pearson correlation analysis. Results: Thirty metabolites were increased significantly, and 5 metabolites were decreased. The differential metabolites are mainly enriched in the metabolism of arachidonic acid, glycero phospholipid, linoleic acid, and thiamine. These results observed that lysophosphatidylcholine (20:1/0:0) and phosphatidylglycerol(PGF1α/16:0) had a tremendous anticipating power with relatively increased AUC values (AUC > 0.9), and they also presented a significant correlation of Mo concentrations in whole blood and Cu concentrations in urine, respectively. Conclusion: The serum metabolism was changed significantly after exposure to welding fume. Lysophosphatidylcholine (20:1/0:0) and phosphatidylglycerol (PGF1α/16:0) may be a potential biological mediator and biomarker for laborers exposure to welding fume. [ABSTRACT FROM AUTHOR]

Artificial oocyte activation (AOA) is considered an effective method to improve clinical outcomes in patients with some forms of male factor infertility and does not increase the risk of birth defects. However, the effects of AOA on patients with multiple morphological abnormalities of the sperm flagella (MMAF) caused by a DNAH1 mutation are still unknown. To explore the effects, our study analyzed a case with MMAF due to DNAH1 homozygous mutation that underwent testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection (ICSI). The case had 28 MII oocytes. The 28 oocytes were divided randomly and equally into AOA and non-AOA groups. Ionomycin was used for AOA. We compared the clinical outcomes of two groups and selected three blastulation failure embryos from each group for transcriptome analysis (Data can be accessed through GSE216618). Differentially expressed genes (DEGs) were determined with an adjusted p-value <0.05 and a |log2-fold change| ≥1. The comparison of clinical outcomes showed that the two pronuclei (2PN) rate and grade 1–2 embryo rate at day 3 were not significantly different between the two groups. Transcriptome analyses of blastulation failed embryos showed that the use of AOA had potential risks of chromosome structure defects, transcriptional regulation defects, and epigenetic defects. In conclusion, when the case with MMAF due to DNAH1 mutation underwent TESE-ICSI, ionomycin-induced oocyte activation could not improve the clinical outcomes and introduced the risks of chromosome structure defect, transcriptional regulation defect, and epigenetic defect. [ABSTRACT FROM AUTHOR]

Epstein-Barr virus-encoded latent membrane protein-1 (LMP1) plays a fundamental role in the malignant transformation of nasopharyngeal carcinoma (NPC), although the mechanism is not well understood. Here, we showed that Limb-bud and Heart (LBH) is considerably downregulated in patient NPC tissues. The expression of LBH in biopsies of 40 consecutive NPC patients devoid of initial distant metastasis and treated according to consistent guidelines was also analyzed, and we found the LBH expression level was correlated with some of clinicopathological features, disease-specific survival (DSS), distant metastasis-free survival (DMFS). We further determined that LBH normally induces NPC cell cycle arrest at the G1/S transition, and LBH can suppress the growth of transplanted NPC tumors in vivo by downregulating LMP1-mediated NF-κB transcriptional activity. Transforming growth factor-beta 1 (TGF-β1) normally protects against tumor development by suppressing cell proliferation, but NPC cells acquire resistance to TGF-β1-mediated inhibition. We found that TGF-β1 inhibits NF-κB transcriptional activity and nasopharyngeal epithelial cell proliferation through upregulating LBH expression. These data reveal a previously unknown NPC transformation mechanism and provide a new concept and treatment strategy for LMP1-driven oncogenesis in NPC. [ABSTRACT FROM AUTHOR]

IgG4-related interstitial lung disease (IgG4-RILD), which is characterized by increased IgG4 levels, IgG4+ plasma cell infiltration and irregular whorled fibrosis, is a recently described lung disorder that belongs to the group of systemic fibroinflammatory IgG4-related diseases (IgG4-RD). The aim of the present study was to improve the current knowledge regarding the specific clinical and histopathological characteristics of IgG4-RILD and to investigate its underlying immune mechanism in vivo. A total of 7 patients newly diagnosed with IgG4-RILD were enrolled in the present study (4 men and 3 women; mean age, 57 years; range, 29-71 years). Patients' clinical history was collected and serological indicators, including C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), antinuclear antibodies (ANAs) and tumor markers were measured. Serum immunoglobulin G (IgG), IgE and IgG4 levels were also evaluated. In addition, computed tomographic (CT) images and pathological examinations were used to determine the characteristics of lung lesions in all patients. The majority of patients presented with symptoms of fever, cough and dyspnea, while allergic symptoms were also encountered. The laboratory examination results revealed different degrees of increased CRP, ESR, tumor markers, ANA, serum IgE and IgG4. The CT images revealed diffuse ground glass opacities, bronchiectasis and thickened bronchovascular bundles. Histologically, the lung lesions were characterized by dense IgG4+ lymphoplasmacytic infiltrates intermixed with extensive fibrous tissue hyperplasia and an irregularly storiform pattern of fibrosis. The mean number of IgG4+ plasma cells was >10 cells/high power field. The ratio of IgG/IgG4+ plasma cells was >50% in inflamed lesions and the number of parenchymal cells was markedly reduced. Obliterative phlebitis or obliterative arteritis was observed in all patients. In conclusion, the clinicopathological similarities between IgG4-RILD and other IgG4-RD suggest that IgG4-related immunopathological processes may be associated with the pathogenesis of pulmonary lesions. Future studies based on the findings herein may elucidate the specific pathological process underlying the development of this fibroinflammatory disorder. [ABSTRACT FROM AUTHOR]

The clinical translation of therapeutic peptides is generally challenged by multiple issues involving absorption, distribution, metabolism and excretion. In this study, a macrophage membrane-coated poly(lactic-co-glycolic acid) (PLGA) nanodelivery system was developed to enhance the bioavailability of the somatostatin (SST) peptide, which faces the hurdles of short half-life and potential side effects in the treatment of chronic pancreatitis. Using a facile nanoprecipitation strategy, SST was loaded in the nanoparticles with an encapsulation efficiency (EE) and a loading efficiency (LE) of 73.68 ± 3.56% and 1.47 ± 0.07%, respectively. The final formulation of SST-loaded nanoparticles with the camouflage of macrophage membrane (MP-SST) showed a mean diameter of 151 ± 4 nm and an average zeta potential of −29.6 ± 0.3 mV, which were stable long term during storage. With an above 90% cell viability, a hemolysis level of about 2% (<5%) and a preference for being ingested by activated endothelial cells compared to macrophages, the membrane–polymer hybrid nanoparticle showed biocompatibility and targeting capability in vitro. After being intravenously administered to mice with chronic pancreatitis, the MP-SST increased the content of SST in the serum (123.6 ± 13.6 pg/mL) and pancreas (1144.9 ± 206.2 pg/g) compared to the treatment of (Dulbecco's phosphate-buffered saline) DPBS (61.7 ± 6.0 pg/mL in serum and 740.2 ± 172.4 pg/g in the pancreas). The recovery of SST by MP-SST downregulated the expressions of chronic pancreatitis-related factors and alleviated the histologic severity of the pancreas to the greatest extent compared to other treatment groups. This augmentation of SST therapeutic effects demonstrated the superiority of integrating the synthetic polymer with biological membranes in the design of nanoplatforms for advanced and smart peptide delivery. Other peptides like SST can also be delivered via the membrane–polymer hybrid nanosystem for the treatment of diseases, broadening and promoting the potential clinical applications of peptides as therapeutics. [ABSTRACT FROM AUTHOR]

The activation of cardiac fibroblasts (CFs) after myocardial infarction (MI) is essential for post-MI infarct healing, during which the regulation of transforming growth factor beta1 (TGF-β1) signaling is predominant. We have demonstrated that TGF-β1-mediated upregulation of LBH contributes to post-MI CF activation via modulating αB-crystallin (CRYAB), after being upregulated by TGF-β1. In this study, the effect of LBH-CRYAB signaling on the cardiac microenvironment via exosome communication and the corresponding mechanisms were investigated. The upregulation of LBH and CRYAB was verified in both cardiomyocytes (CMs) and CFs in hypoxic, post-MI peri-infarct tissues. CM-derived exosomes were isolated and identified, and LBH distribution was elevated in exosomes derived from LBH-upregulated CMs under hypoxia. Treatment with LBH+ exosomes promoted cellular proliferation, differentiation, and epithelial-mesenchymal transition-like processes in CFs. Additionally, in primary LBHKO CFs, western blotting showed that LBH knockout partially inhibited TGF-β1-induced CF activation, while LBH-CRYAB signaling affected TGF-β1 expression and secretion through a positive feedback loop. The administration of a Smad3 phosphorylation inhibitor to LBHKO CFs under TGF-β1 stimulation indicated that Smad3 phosphorylation partially accounted for TGF-β1-induced LBH upregulation. In conclusion, LBH upregulation in CMs in post-MI peri-infarct areas correlated with a hypoxic cardiac microenvironment and led to elevated exosomal LBH levels, promoting the activation of recipient CFs, which brings new insights into the studies and therapeutic strategies of post-MI cardiac repair. [ABSTRACT FROM AUTHOR]

(E) Immunohistochemical analysis of the expression of CD20 in serial sections of IgG4-related lung disease. Graph: Figure 3 Immunohistochemical analysis of the expression of (A) CD68 in serial sections of IgG4-related lung disease. (F) Immunohistochemical analysis of the expressions of CD38 in serial sections of IgG4-related lung disease. [Extracted from the article]

Inflammatory bowel disease (IBD) is a group of immune-mediated conditions. Immune activity is varied by age and gender. The present study is aimed at investigating the effect of age and gender on the positive rates of anti-Saccharomyces cerevisiae antibodies (ASCA), anti-neutrophil cytoplasmic antibodies (ANCA), anti-intestinal goblet cell antibodies (GAB), and antibodies to exocrine pancreas (PAB) in IBD patients. A total of 1871 hospitalized patients with confirmed IBD were included in this study. Sera were obtained from each subject for antibody measurement by indirect immunofluorescence assay. The positive rates of ANCA IgG and IgA were higher in female patients than those in male patients (P < 0.001) while the positive rate of PAB IgG was just reversed (P < 0.001). Moreover, the median ages of patients with positive ANCA IgG and IgA were higher than patients with negative antibodies (P = 0.0019 and P = 0.0110 , respectively), while the median ages of patients with positive PAB IgG and IgA were significantly lower than patients with negative PAB (P < 0.0001). The serum levels of ANCA IgG and IgA were potentiated in old female patients, while serum PAB IgG was easy to be detected in the young male patients with IBD. [ABSTRACT FROM AUTHOR]

Objectives. To investigate the relationship between polymorphisms of calcitonin-related peptide gene II (beta-calcitonin gene-related peptide (βCGRP), CALCB) and serum CGRP levels in salivary adenoid cystic carcinoma. Materials and Methods. Using the polymerase chain reaction (PCR) technique, the full-length amplification and genotype analysis of CALCB genes were performed in 39 patients with adenoid cystic carcinoma of salivary gland and 158 normal controls. The gene frequencies of major genotype of CALCB in adenoid cystic carcinoma of salivary gland and normal control group were analyzed. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate serum calcitonin gene-related peptide (CGRP) and its concentration of alpha and beta subtypes. Results. Univariate logistic regression analysis showed that the CALCB rs2839222 T/T genotype was closely related to the occurrence of salivary adenoid cystic carcinoma, with a correlation coefficient of 3.89. Conclusions. The serum CGRP concentration in the salivary adenoid cystic carcinoma group was 1.56 times that of the normal control group. The αCGRP subtype was significant, which was 3.02 times that of the normal control. The polymorphism of βCGRP gene is associated with genetic susceptibility to salivary adenoid cystic carcinoma, and serum CGRP and βCGRP can be used as novel markers of salivary adenoid cystic carcinoma. [ABSTRACT FROM AUTHOR]

Pancreatic cancer is one of the most common malignancies worldwide. This study is aimed at searching the possible genetic mutations and the value of novel gene mutation in the DNA damage-inducible transcript 4 (DDIT4) and signaling pathway in pancreatic cancer. Polymerase chain reaction (PCR) was performed to amplify the DNA sequences of DDIT4 from patients with pancreatic ductal adenocarcinoma. In addition, we used IHC to detect the expression level of DDIT4 in patients with pancreatic cancer in different types of gene mutation. Double-labeled immunofluorescence was employed to explore the expression levels of DDIT4/LC3 and their potential correlation. Our work indicated the two novel stable gene mutations in DDIT4 mRNA 3 ′ -untranslated region (m.990 U>A and m.1246 C>U). Thirteen samples were found to have mutation in the DDIT4 3 ′ -untranslated regions (UTR). To further verify the influence of gene mutation on protein expression, we performed immunohistochemistry on different gene mutation types, and we found a correlation between DDIT4 expression and gene mutation, which is accompanied by nuclear staining deepening. In order to further discuss the clinical value of DDIT4 gene mutation, immunofluorescence suggested that the expression of DDIT4 colocated with LC3; thus, we speculated that DDIT4 mutation may be involved in autophagy in pancreatic cancer cell. In this study, we found mutation in the 3 ′ -UTR region of DDIT4, which may be associated with DDIT4 expression and tumor autophagy in pancreatic cancer tissues. [ABSTRACT FROM AUTHOR]

DDIT4 is a mitochondrial and tumor-related protein involved in anti-tumor therapy resistance, proliferation, and invasion, etc. Its expression level increases under the stress such as chemotherapy, hypoxia, and DNA damage. A number of clinical studies have confirmed that DDIT4 can change the behavior of tumor cells and the prognosis of patients with cancer. However, the role of DDIT4 in promoting or suppressing cancer is still inconclusive. This article summarized the four characteristics of DDIT4 including a mitochondria-related protein, interactions with various protein molecules, immune and metabolic cell related proteins and participator in the oxygen sensing pathway, which may be related to the progress of cancer. [ABSTRACT FROM AUTHOR]

Amentoflavone, robustaflavone, 2″,3″-dihydro-3′,3‴-biapigenin, 3′,3‴-binaringenin, and delicaflavone are five major hydrophobic components in the total biflavonoids extract from Selaginella doederleinii (TBESD) that display favorable anticancer properties. The purpose of this study was to develop a new oral delivery formulation to improve the solubilities, dissolution rates, and oral bioavailabilities of the main ingredients in TBESD by the solid dispersion technique. Solid dispersions of TBESD with various hydrophilic polymers were prepared, and different technologies were applied to select the suitable carrier and method. TBESD amorphous solid dispersion (TBESD-ASD) with polyvinylpyrrolidone K-30 was successfully prepared by the solvent evaporation method. The physicochemical properties of TBESD-ASD were investigated by scanning electron microscopy, differential scanning calorimetry, and Fourier-transform infrared spectroscopy. As a result, TBESD was found to be molecularly dispersed in the amorphous carrier. The solubilities and dissolution rates of all five ingredients in the TBESD-ASD were significantly increased (nearly 100% release), compared with raw TBESD. Meanwhile, TBESD-ASD showed good preservation stability for 3 months under accelerated conditions of 40 °C and 75% relative humidity. A subsequent pharmacokinetic study in rats revealed that Cmax and AUC0–t of all five components were significantly increased by the solid dispersion preparation. An in vivo study clearly revealed that compared to raw TBESD, a significant reduction in tumor size and microvascular density occurred after oral administration of TBESD-ASD to xenograft-bearing tumor mice. Collectively, the developed TBESD-ASD with the improved solubility, dissolution rates and oral bio-availabilities of the main ingredients could be a promising chemotherapeutic agent for cancer treatment. [ABSTRACT FROM AUTHOR]

In this work, quaternary ammonium carbon quantum dots (QCQD) was successfully synthesized with 2,3-epoxypropyltrimethylammonium chloride and diallyldimethylammonium chloride by a simple green "one-pot" method. QCQD was demonstrated to have satisfactory antibacterial activity against gram-positive bacteria, including Staphylococcus aureus , methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis , Listera monocytogenes and Enterococcus faecalis. QCQD was used to treat MRSA-infected pneumonia in mice, and could promote the regression of pulmonary inflammation in lung of mice. In vivo and in vitro toxicity tests also confirmed that QCQD had acceptable biocompatibility. There is no doubt that all the experimental results confirmed that QCQD is a promising antimicrobial agent for treating infections of gram-positive bacteria. The antibacterial mechanism of QCQD was analyzed and confirmed by the quantitative proteomics based tandem mass tags, revealing that QCQD might interfere with the protein translation, posttranslational modification and protein turnover in bacterial cells by mainly acting on ribosomes and upregulating RNA degradation related proteins. The application of proteomics extended the research on antibacterial mechanism from single target to protein network and realized the research and analysis on multiple targets of bacteria treated by QCQD, providing a new idea and strategy for the development of efficient bactericidal weapon against drug-resistant bacteria. Image 1 [ABSTRACT FROM AUTHOR]

This study explored the triggering mechanism of interstitial lung disease (ILD). We established the effects of immunogenic and neurogenic calcitonin gene-related peptide (CGRP) imbalance on the regulation of aquaporin 5 (AQP5) expression and the repair responses that promote the transition from alveolar epithelial cell (AEC) apoptosis to pulmonary fibrosis. Newly diagnosed ILD patients (n = 60) were enrolled, whose serological levels of β-CGRP, α-CGRP, AQP5, receptor activity modifying protein 1, and receptor component protein were detected by ELISA. Th1 and Th2 cytokines and CD4+ and CD8+ cells were measured by flow cytometry method. In vivo, bleomycin (BLM) was set for modeling pulmonary fibrosis. A CALCA-HET model was set as a chronic pulmonary fibrosis model. Hematoxylin–eosin, immunohistochemistry, and Masson's trichrome staining were performed to assess the role of apoptosis in the injured lung. The concentrations of cytokines were determined by cytokine antibody arrays. Abnormal activation of serological CD4+ T lymphocytes and predominant Th2 response was established in the patients with ILD. Moreover, the ratio of β-CGRP/α-CGRP positively correlated with the increased level of AQP5 in patients with ILD. In vivo, a significant increase of AQP5 and β-CGRP at the chronic stage of pulmonary fibrosis was induced by BLM in the mice model, whereas the expression of AQP5 protein was generally lower in the acute alveolitis phase. Moreover, the levels of AQP5 and α-CGRP in the CALCA-HET rats were lower than those of the normal saline group. The high ratio β-CGRP/α-CGRP enhanced the expression of AQP5, inhibited transforming growth factor-β1 (TGFβ1)/P-Smad1/Smad4 pathway, and upregulated the NRF2 signal, whereas the apoptosis of AECs was significantly reduced in late-stage pulmonary fibrosis. The imbalance of β-CGRP/α-CGRP may be associated with the predominance of Th2 response and participate in the process of AEC apoptosis in lung fibrosis. The high ratio of β-CGRP/α-CGRP may elevate the level of AQP5 through inactivation of the TGF-β1/smad1 signaling pathway and upregulation of the Nrf2 signaling in the chronic stage of pulmonary fibrosis. [ABSTRACT FROM AUTHOR]

The prognostic value of the genotype of the PRSS1 gene in patients with pancreatic ductal adenocarcinoma (PDAC) remains poorly understood. The aim of the present study was to evaluate the association between the PRSS1 genotype and clinicopathological characteristics of patients with PDAC, as well as to explore the prognostic significance of the PRSS1 genotype in patients with PDAC. A total of 124 patients with PDAC patients were included in the current study and the PRSS1 genotype of the enrolled patients was determined by the polymerase chain reaction. Associations between the PRSS1 genotype and clinicopathological characteristics were subsequently analyzed using the Chi-square test. The impact of the PRSS1 genotype on patient prognosis was assessed using the Kaplan-Meier method, and predictive factors of overall survival (OS) time were analyzed by Cox regression. A total of 56 patients with PDAC (45.16%) had the T/C PRSS1 genotype, which was associated with large tumor sizes (P=0.027) and higher tumor node metastasis (TNM) stages (P=0.041). Following a median follow-up of 19 months, the T/C genotype of PRSS1 genotype was associated with a shorter OS time (P=0.037) compared with the C/C or T/T PRSS1 genotypes. Univariate and multivariate analyses revealed that PRSS1 genotype was identified to be an independent prognostic factor for the OS time of patients with PDAC. The results obtained in the current study suggested that the PRSS1 genotype, as well as factors such as the serum level of carbohydrate antigen 19-9 and the TNM stage, may act as independent prognostic factors for the OS time of patients with PDAC. [ABSTRACT FROM AUTHOR]

Corona Virus Disease 2019 (COVID-19) is an acute respiratory infection and a global public health event. The level of aβ 2 GPI is significantly up-regulated in COVID-19 patients. The impact of inactivated vaccination against COVID-19 on aβ 2 GPI and in vitro fertilization and embryo transfer (IVF-ET) remains unknown amidst the universal administration of COVID-19 vaccines. We conducted a retrospective study to assess the impact of COVID-19 inactivated vaccination on aβ 2 GPI levels and its effect on superovulation and pregnancy outcomes. We found aβ 2 GPI level is significantly up-regulated after vaccination. There was no statistical difference in mature egg rate, 2PN fertilization rate, day 3 high-quality embryo rate, blastocyst formation rate, embryo implantation rate and miscarriage rate between the vaccine group and control group. Our findings showed vaccination with COVID-19 inactivated vaccine can elevate the level of aβ 2 GPI in peripheral blood but have no effect on the outcomes of controlled ovarian hyperstimulation and pregnancy in IVF-ET. • This is the first study to explore the correlation between the level of aβ 2 GPI in patients after inactivated COVID-19 vaccination and the outcome of ovulation hyperstimulation and pregnancy in IVF-ET. • The results showed aβ 2 GPI level is significantly up-regulated after inactivated COVID-19 vaccination. • The results showed no statistical difference in mature egg rate, 2PN fertilization rate, day 3 high-quality embryo rate, blastocyst formation rate, embryo implantation rate and miscarriage rate betwee n the vaccine group and control group. [ABSTRACT FROM AUTHOR]

Abstract: Sarcosine is a potential prostate cancer marker. In this study, we developed a method of three‐phase solvent bar liquid‐phase microextraction combined with high‐performance liquid chromatography to determine sarcosine after derivatization with 4‐dimethylarminoazobenzene‐4‐sulfonyl chloride from human urine. The effects of different extraction conditions on extraction efficiency were investigated and optimized. Under optimum experimental conditions, a calibration graph exhibited linearity over the range of 0.05–25 μmol/L with a correlation coefficient (r2) of 0.9990. The enrichment factor was 168, and the detection limit was 0.02 μmol/L. The method was successfully used to analyze sarcosine in human urine and non‐invasive detection, and good spiked recoveries ranging from 90.5 to 93.6% were obtained. The proposed method exhibited high sensitivity, high enrichment factor, good precision, and a simple setup. It may contribute to the early accurate diagnosis and the progression monitoring of prostatic carcinoma. [ABSTRACT FROM AUTHOR]

α-Synuclein phosphorylation and mitochondrial calcium homeostasis are important mechanisms underlying mitochondrial dysfunction in Parkinson's disease, but the network regulating these mechanisms remains unclear. We identified the role of key phosphokinases and the pathological effects of α-synuclein phosphorylation on mitochondrial calcium influx and mitochondrial function in Parkinson's disease. The function of the key phosphokinase, calcium/calmodulin-dependent serine protein kinase, was investigated through loss- and gain-of-function experiments using a cell model of Parkinson's disease. The regulation of mitochondrial calcium uniporter-mediated mitochondrial calcium influx by calcium/calmodulin-dependent serine protein kinase was explored using a cellular model of Parkinson's disease. Coimmunoprecipitation experiments and α-synuclein mutation were used to explore the mechanism through which calcium/calmodulin-dependent serine protein kinase regulates mitochondrial calcium uniporter-mediated mitochondrial calcium influx and exacerbates mitochondrial damage in Parkinson's disease. Here, we show the pathogenic role of calcium/calmodulin-dependent serine protein kinase in Parkinson's disease progression. Calcium/calmodulin-dependent serine protein kinase phosphorylated α-synuclein to activate mitochondrial calcium uniporter and thus increase mitochondrial calcium influx, and these effects were blocked by α-synuclein S129A mutant expression. Furthermore, the calcium/calmodulin-dependent serine protein kinase inhibitor CASK-IN-1 exerted neuroprotective effects in Parkinson's disease. Collectively, our results suggest that calcium/calmodulin-dependent serine protein kinase phosphorylates α-synuclein to activate the mitochondrial calcium uniporter and thereby causes mitochondrial calcium overload and mitochondrial damage in Parkinson's disease. We elucidated a new role of calcium/calmodulin-dependent serine protein kinase in Parkinson's disease and revealed the potential therapeutic value of targeting calcium/calmodulin-dependent serine protein kinase in Parkinson's disease treatment. [Display omitted] • CASK was firstly verified to be upregulated during PD progression. • CASK activated p-a-syn-MCU signaling to modulate mitochondrial calcium overload. • CASK could serve as a candidate target among the future development of PD treatment. [ABSTRACT FROM AUTHOR]

Human neuropeptide Y (hNPY) is one of the most widely expressed neurotransmitters in the human central and peripheral nervous systems. It consists of 36 highly conserved amino acid residues, and was first isolated from the porcine hypothalamus in 1982. While it is the most recently discovered member of the pancreatic polypeptide family (which includes neuropeptide Y, gut-derived hormone peptide YY, and pancreatic polypeptide), NPY is the most abundant peptide found in the mammalian brain. In order to exert particular functions, NPY needs to bind to the NPY receptor to activate specific signaling pathways. NPY receptors belong to the class A or rhodopsin-like G-protein coupled receptor (GPCR) family and signal via cell-surface receptors. By binding to GPCRs, NPY plays a crucial role in various biological processes, including cortical excitability, stress response, food intake, circadian rhythms, and cardiovascular function. Abnormal regulation of NPY is involved in the development of a wide range of diseases, including obesity, hypertension, atherosclerosis, epilepsy, metabolic disorders, and many cancers. Thus far, five receptors have been cloned from mammals (Y1, Y2, Y4, Y5, and y6), but only four of these (hY1, hY2, hY4, and hY5) are functional in humans. In this review, we summarize the structural characteristics of human NPY receptors and their role in metabolic diseases. [ABSTRACT FROM AUTHOR]

Searching for potential anticancer agents from natural sources is an effective strategy for developing novel chemotherapeutic agents. In this study, data supporting the in vitro and in vivo anticancer effects of delicaflavone, a rarely occurring biflavonoid from Selaginella doederleinii, were reported. Delicaflavone exhibited favorable anticancer properties, as shown by the MTT assay and xenograft model of human non-small cell lung cancer in male BALB/c nude mice without observable adverse effect. By transmission electron microscopy with acridine orange and Cyto-ID®Autophagy detection dyes, Western blot analysis, and RT-PCR assay, we confirmed that delicaflavone induces autophagic cell death by increasing the ratio of LC3-II to LC3-I, which are autophagy-related proteins, and promoting the generation of acidic vesicular organelles and autolysosomes in the cytoplasm of human lung cancer A549 and PC-9 cells in a time- and dose-dependent manner. Delicaflavone downregulated the expression of phospho-Akt, phospho-mTOR, and phospho-p70S6K in a time- and dose-dependent manner, suggesting that it induced autophagy by inhibiting the Akt/mTOR/p70S6K pathway in A549 and PC-9 cells. Delicaflavone is a potential anticancer agent that can induce autophagic cell death in human non-small cell lung cancer via the Akt/mTOR/p70S6K signaling pathway. Delicaflavone showed anti-lung cancer effects in vitro and in vivo. Delicaflavone induced autophagic cell death via Akt/mTOR/p70S6K signaling pathway. Delicaflavone did not show observable side effects in a xenograft mouse model. Delicaflavone may represent a potential therapeutic agent for lung cancer. Key messages: [ABSTRACT FROM AUTHOR]

In this work, a signal-off double probes electrochemical DNA sensor was developed to simultaneously detect Legionella and Legionella pneumophila . We designed hairpin probes which modified by ferrocene/methylene blue (Fc/MB) for selective electrochemical detecting Legionella and Legionella pneumophila . The hairpin probes were immobilized by Au-S chemical bond reaction on the surface of AuE electrodes, which can be cleaved by EcoRI in the absence of Legionella, but cannot be cleaved in the presence of Legionella or Legionella pneumophila . The current changes before and after DNA hybridization and enzymatic cleavage were monitored by the proposed DNA sensor with differential pulse voltammetry. Electrochemical signals was generated by Fc/MB. There was a good linear relationship between the peak current value and the logarithmic concentration of Legionella or Legionella pneumophila . The linear regression equation of Legionella was I (nA) = − 111.57log C (mol/L) + 1084.0, R 2 = 0.9902; and the linear regression equation of Legionella pneumophila was I (nA) = − 126.09log C (mol/L) + 1238.2, R 2 = 0.9945. The linear range had been estimated from 2.0 × 10 − 9 to 2.0 × 10 − 4 mol/L. Legionella , Legionella pneumophila and other bacteria were sensitively distinguished by the proposed DNA sensor. And the sensor could be used for detecting the Legionella pneumophila samples from patients or environments. [ABSTRACT FROM AUTHOR]

Trypsin is a protease-activated receptor-2 (PAR2) activator that upregulates the interleukin (IL)-17 receptor signal in the airway epithelial cells and amplifies the inflammatory response, but does not modify the growth kinetics of the metapneumovirus. How does the coronavirus spread from cell to cell is yet an enigma. The present study analyzed the possible role of trypsin in the activation of coronavirus in vitro and in vivo. We found that the overexpression of trypsin in A549 cells upregulated IL-17 and angiotensin-converting enzyme (ACE). In the humanized transgenic mice, trypsin activated M1 macrophages. Together, our results suggested that the upregulation of trypsin may support a new pathway for coronavirus transmission in patients. • The spike protein of CoV is cleaved by trypsin, cathepsin and other active proteases and activated for fusion. • The cutting event of CoV leads to the fusion of cells and virus. • Trypsin may play an important role in the process of virus aggregation, release and replication. [ABSTRACT FROM AUTHOR]

We report a novel method for the amplified detection of K-ras point mutation by using a chronocoulometric DNA sensor based on Escherichia coli DNA ligase and AuNPs amplification effects. Our method is based on a ‘sandwich’ detection strategy, involving capture probe immobilized on a gold electrode (GE) and reporter probe loaded on Au nanoparticles (AuNPs probe). Each probe flanks one of two fragments of the target DNA sequence. Successive hybridization event brings AuNPs, along with hundreds of reporter probes to form the ‘sandwich’ structure. After high-fidelity E. coli DNA ligase reaction and denaturing at an elevated temperature, We then employ chronocoulometry to detect [Ru(NH 3 ) 6 ] 3+ electrostatically bounding to the DNA probe strands. If there is an allele mismatch, no ligation takes place. In this way, the novel approach had successfully distinguish the K-ras mutant DNA and the K-ras wild DNA. The charge variable (Δ Q ) is proportional to the logarithmic K-ras mutant DNA concentration from 1.0 nM to 0.1 pM, with the detection limit of 0.01 pM. Owing to its ease of operation and low detection limit, it is expected that the proposed approach may hold great promise in both research-based and clinical genomic assays. [ABSTRACT FROM AUTHOR]

Neonatal sepsis remains an important and common cause of morbidity and mortality among newborn infants, especially in developing countries. The aim of the present study was to determine whether serum trypsin levels and genotypes of cationic trypsinogen ( PRSS1) gene could be served as markers for predicting neonatal sepsis. The serum trypsin levels and genotypes of PRSS1 were examined in both 50 infants with infection during neonatal period and 56 healthy neonates as controls. The infected infants were further subdivided into infants with sepsis group ( n = 18) and infected infants without sepsis ( n = 32). The genotype of PRSS1 was analyzed by direct sequencing, and the serum trypsin level was measured by immunoassay. It showed that the median value of serum trypsin was significantly higher in infected infants (31.90 ng/mL) than in controls (12.85 ng/mL; P = 0.030). More importantly, sepsis subgroup (50.95 ng/mL) had significantly higher median serum trypsin than infected infants without sepsis subgroup (19.10 ng/mL) and controls (12.85 ng/mL) ( P = 0.015 and P = 0.002, respectively). Additionally, the median serum trypsin levels were found significantly higher in infants who had T/T (37.90 ng/mL) genotype of PRSS1 compared with those who had C/T genotype (12.80 ng/mL; P = 0.005). This study suggested that serum trypsin and rs10273639 C/T of PRSS1 were revealed to be novel markers for predicting neonatal sepsis. [ABSTRACT FROM AUTHOR]

[Display omitted] • 4′''ADLF was synthesized and evaluated for anticancer activities against 10 cancer cells. • 4′''ADLF achieves better antitumor activity against HepG2 in vivo and in vitro, and induces apoptosis and arrests cell cycle. • Improvements in pharmacokinetics of 4′''ADLF compared with DLF. • Network pharmacology of 4′''ADLF and molecular docking of six high score proteins are both discussed. • 4′''DLF induced autophagy mainly by activating MAPK phosphorylation pathway with a dose-dependent inhibitory effect. Structural modification of natural products is the effective option to improve their pharmacological effects and drug properties. DLF is a lead compound of antitumor drug, which is a broad-spectrum, low toxic and high-efficient component isolated from Selaginella doederleinii Hieron by our research group. Here, we report the structural modification method of this component, and find that the acetylated product of C4′′′- OH (C4′′′-acetyl-delicaflavone, 4′′′ADLF) has better inhibitory effect on the selected cancer cell lines, including, lung, liver, colon and cervical cancer cell lines. Since the increased water solubility of 4′′′ADLF may lead to higher absorption rate and activity, we evaluate the pharmacodynamics in vitro and in vivo, and the pharmacokinetic of 4′′′ADLF. It shows that 4′′′ADLF inhibit the proliferation and induce cycle arrest in tumor cells, and had better anticancer activity and bioavailability than DLF. [ABSTRACT FROM AUTHOR]

The venoms of toxic animals are chemical pools composed of various proteins, peptides, and small organic molecules used for predation and defense, in which the peptidic toxins have been intensively pursued mining modulators targeting disease-related ion channels and receptors as valuable drug pioneers. In the present study, we uncovered the molecular diversity of peptide toxins in the venom of the spider Heteropoda pingtungensis (H. pingtungensis) by using a combinatory strategy of venom gland cDNA library and transcriptome sequencing (RNA-seq). An amount of 991 high-quality expressed sequence tags (ESTs) were identified from 1138 generated sequences, which fall into three categories, such as the toxin-like ESTs (531, 53.58%), the cellular component ESTs (255, 25.73%), and the no-match ESTs (205, 20.69%), as determined by gene function annotations. Of them, 190 non-redundant toxin-like peptides were identified and can be artificially grouped into 13 families based on their sequence homology and cysteine frameworks (families A–M). The predicted mature toxins contain 2–10 cysteines, which are predicted to form intramolecular disulfide bonds to stabilize their three-dimensional structures. Bioinformatics analysis showed that toxins from H. pingtungensis venom have high sequences variability and the biological targets for most toxins are unpredictable due to lack of homology to toxins with known functions in the database. Furthermore, RP-HPLC and MALDI-TOF analyses have identified a total of 110 different peptides physically existing in the H. pingtungensis venom, and many RP-HPLC fractions showed potent inhibitory activity on the heterologously expressed NaV1.7 channel. Most importantly, two novel NaV1.7 peptide antagonists, µ-Sparatoxin-Hp1 and µ-Sparatoxin-Hp2, were characterized. In conclusion, the present study has added many new members to the spider toxin superfamily and built the foundation for identifying novel modulators targeting ion channels in the H. pingtungensis venom. [ABSTRACT FROM AUTHOR]

The fabrication of a facile, sensitive, and versatile immunosensor for the quantification of metallothionein-3 (MT-3) is proposed in this work. The K3[Fe(CN)6]–chitosan–glutaraldehyde (K–CS–GA) conjugate prepared from K3[Fe(CN)6], chitosan and glutaraldehyde was employed as the redox-active signal source. Carbon nanodots (C-dots) were coupled with Nafion to form the nanocomposite architecture layer to carry antibodies (Abs). C-dots enhanced the electrochemical response of the proposed immunosensor to improve the detection sensitivity. The fabrication steps of the immunosensor were characterized using differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Antigen determination was achieved via the decreased current response of the K3[Fe(CN)6] caused by the insulated coupled antigen. The detected signals were proportional to the logarithm of the concentrations of MT-3 ranging from 5 pg mL−1 to 20 ng mL−1 with a detection limit of 2.5 pg mL−1 in PBS. The proposed immunosensor showed high sensitivity, good selectivity and reproducibility. Furthermore, detection results using real serum samples showed the immunosensor's potential applications in clinical diagnostics. [ABSTRACT FROM AUTHOR]

Abstract: This study reports on a novel biosensor for K-ras mutation gene detection by using hairpin locked nucleic acids (LNA) probes dually labeled with amines groups and biotin. The hairpin LNA probes was covalently immobilized on poly-Eriochrome cyanine R (ECR) film modified glassy carbon electrode (GCE) as a switch. Streptavidin-horseradish Peroxidase labeled gold nanoparticles (SA-HRP-AuNPs) bioconjugate were used as tracer for signal amplification. The immobilized hairpin LNA probe was in the “closed” state in the absence of K-ras mutation gene, which shielded biotin from being approached by the bulky SA-HRP-AuNPs bioconjugate due to the steric effect. When hairpin LNA probes hybridized with K-ras mutation gene, hairpin LNA probes undergo significant conformational change, forcing biotin away from GCE surface. As a result, the biotin label becomes accessible by the SA-HRP-AuNPs bioconjugate, and K-ras mutation gene hybridization event can be sensitively transduced via the numerous HRP enzymatically amplified electrochemical current signal. This new biosensor had a good specificity to distinguish the K-ras mutations, K-ras wild-type sequence, and a high sensitivity. [Copyright &y& Elsevier]

Abstract: We herein proposed a novel, facile, and sensitive strategy for selective detection of tumor necrosis factor-alpha (TNF-α) using K3[Fe(CN)6] as the label-free signal. A K3[Fe(CN)6]-chitosan-glutaraldehyde system (K-CS-GA) capable of high signal indication was developed for the first time to improve the sensitivity of the electrochemical immunosensor. K-CS-GA acting as the signal indicator was first fixed on a glass carbon electrode to be the signal layer. Subsequently, nafion was used to cover the signal layer and immobilize mouse anti-human TNF-α onto the electrode. The target TNF-α captured onto the electrode through immunoreactions efficiently induced the current response of the electrode due to the non-conducting biomoleculars. TNF-α was detected by monitoring the electrode current response in PBS using cyclic voltammetry (CV), which proved to be sensitive to varying concentrations of TNF-α. The decreases of anodic currents were proportional to the logarithm of the antigen concentrations in the range of 0.02–34ng/ml with the detection limit of 10pg/ml. The proposed immunosensor not only demonstrated high sensitivity, good selectivity, acceptable stability, and reproducibility, but also exhibited potential applications in clinical protein diagnostics. [Copyright &y& Elsevier]

Nanozymes, as a new type of artificial enzyme, have recently become a research hotspot in the field of catalysis and biomedicine. However, the application of nanozyme is limited by catalytic activity changes of different substrates and low specificity. This work shows that citrate-capped platinum nanoparticles (Cit-PtNPs) exhibit stronger oxidase-like activity than other platinum nanozymes at different pH when 3-methyl-2-benzothiazolinonehydrazone hydrochloride (MBTH) and n-ethyl-n- (2-hydroxy-3-sulfopropyl)-m-toluidine sodium salt (TOOS) were used as chromogenic substrates. This phenomenon has important reference value for different nanozymes to choose chromogenic substrates in catalysis. In MBTH-TOOS chromogenic system, MBTH (-NH) radical is first produced during the reaction through catalytic oxidation of Cit-PtNPs, which reacts with TOOS to produce a colorless compound. The blue-purple quinoid dye was produced through the dismutation of the colorless compound. The catalytic mechanism of the oxidase-like activity of Cit-PtNPs is that two-electron reduction process and four-electron reduction process are simultaneously carried out in the catalytic process. Furthermore, to solve the problem of low specificity of metal nanozymes, protamine is designed as aggregation promoter of Cit-PtNPs and the specifichydrolysis substrate of trypsin. In this work, it can achieve one-step detection of trypsin by the boosting oxidase activity of Cit-PtNPs at pH8. The catalytic activity of Cit-PtNPs is proportional to the concentration of trypsin. The linear range for trypsin is 1.0–70.0 ngmL−1 and the limit of detection is measured to be 0.6 ngmL−1. This novel method has also been successfully applied to the detection of inhibitors and trypsin in urine samples. [Display omitted] • Cit-PtNPs with robust oxidase-like activity and a wide pH range in MBTH-TOOS chromogenic system. • Experiments indicates that MBTH (-NH) radical is first produced during the reaction by catalytic oxidation of Cit-PtNPs. • Two-electron and four-electron reduction process are simultaneously carried out in the catalytic process of Cit-PtNPs. • The Cit-PtNPs were applied as a nanoprobe for facile, selective, and fast colorimetric detection of trypsin and inhibitor. [ABSTRACT FROM AUTHOR]

Abstract: An amplified chronocoulometric Locked nucleic acid (LNA) sensor (CLS) for selective electrochemical detection of K-ras mutation was developed based on site-specific DNA cleavage of restriction endonuclease EcoRI. Thiolated-hairpin LNA probe with palindrome structure of stem was immobilized on the gold nanoparticles modified gold electrode (NG/AuE). It can be cleaved by EcoRI in the absence of K-ras mutation-type DNA (complementary with the loop part of hairpin probe), but cannot be cleaved in the presence of mutation-type DNA. The difference before and after enzymatic cleavage was then monitored by chronocoulometric biosensor. Electrochemical signals are generated by chronocoulometric interrogation of Hexaammineruthenium (III) chloride (RuHex) that quantitatively binds to surface-confined hairpin LNA probe via electrostatic interactions. The results suggested this CLS had a good specificity to distinguish the K-ras mutation-type, wild-type and non-complementary sequence. There was a good linear relationship between the charge and the logarithmic function of K-ras mutation-type DNA concentration. The detection limit had been estimated as 0.5fM. It is possible to qualitatively and quantitatively detect K-ras point mutation in pancreatic cancer. [Copyright &y& Elsevier]

Background and Aims: The high mortality rate of pancreatic cancer is a bottleneck for further treatment with long-term efficacy. Thus, it is urgent to identify new methods to accurately predict the early onset of pancreatic cancer. We hypothesized that the different genotypes of cationic trypsinogen (PRSS1) gene could confer susceptibility and/or resistance to pancreatic cancer in the Han Chinese population.Methods: The genotypes of PRSS1 were determined in 154 patients with pancreatic cancer and in a control group of 520 healthy individuals of Han Chinese descent. Clinical information was obtained, single-nucleotide polymorphisms (SNPs) of the PRSS1 gene were analyzed by direct sequencing, and the distribution of the genotypes were tested for Hardy-Weinberg equilibrium. Odds ratios and 95% confidence intervals were calculated by logistic regression analysis to estimate the associations between the different genotypes or haplotypes and the risk of pancreatic cancer.Results: Three SNPs (-409 C/T, -204 A/C, and c.486 C/T) were identified. A case-control analysis revealed a 0.118-fold (95% CI: 0.037-0.653), 0.842-fold (95% CI: 0.177-4.010), and 0.750-fold (95% CI: 0.519-1.085) change in risk of developing pancreatic cancer for individuals harboring these SNPs, respectively. The individuals with the -409 C/T genotype tended to have a reduced risk compared to those who carried the -409 T/T genotype. A protective effect was observed for the C(-409)-A(-204)-C(486) haplotype compared to the T(-409)-A(-204)-T(486) haplotype (OR = 0.115, 95% CI: 0.016-0.849) or compared to the T(-409)-A(-204)-C(486) haplotype (OR = 0.090, 95% CI: 0.012-0.667). Serum levels of trypsin in patients with the -409 C/T genotype were only one-fourth that of those with the -409 T/T genotype and only one-third that of the healthy controls.Conclusions: The -409 C/T genotype of PRSS1 was revealed to be a protective factor against pancreatic cancer in the Han Chinese population. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a kind of nanoporous gold electrode (NPG) prepared with repetitive square-wave oxidation reduction cycle (SWORC) was reported. The active surface area of the proposed NPG electrode was 9.9 times larger than that of a bare flat one characterized by cyclic voltammetry (CV). An electrochemical DNA biosensor based on NPG electrode was fabricated for detection of promyelocytic leukemia/retinoic acid receptor α (PML/RARα) fusion gene in acute promyelocytic leukemia (APL) by using Methylene Blue (MB) as an electroactive indicator. Differential pulse voltammetry (DPV) was employed to monitor the hybridization reaction on the probe modified electrode. The decrease of the peak current of MB was observed upon hybridization of the probe with target DNA. The results indicated that the peak current was linear with the concentration of complementary strand in the range of 60pM to 220pM with a detection limit of 6.7pM. This new biosensor exhibited excellent sensitivity and selectivity and had been used for an assay of PCR real sample with a satisfactory result. [Copyright &y& Elsevier]

Limb-bud and heart (LBH) gene has received increasing attention in recent cancer studies. Here we investigated the role of the LBH gene in regulating the metastasis capacity and epithelial-mesenchymal transition (EMT) of nasopharyngeal carcinoma (NPC) cells, and its potential mechanism. The expressions of LBH and αB-crystallin (CRYAB) were modulated by lentiviral infection, or plasmid/siRNA transfection, and the phosphorylation of p38 was suppressed by an inhibitor, to explore their functions in modulating NPC cell phenotypes, as well as the relationships of these factors with each other. Cellular proliferation, migration and invasion were examined by RTCA system, Transwell assays and Matrigel Transwell assays respectively. The EMT progression was indicated by RT-qPCR and Western blotting measuring the expressions of EMT biomarkers. NPC xenografts were constrcucted, and formed tumors were sectioned for morphology and immunohistofluorescence. The interaction between LBH and CRYAB was examined by colocalization and Fluorescence resonance energy transfer (FRET) analysis. We reached the conclusion that LBH inhibits the proliferation, migration, invasion and EMT of NPC cells, and its effects were partially achieved by suppressing p38 phosphorylation, which subsequently downregulates the mRNA expression and phosphorylation of CRYAB, while CRYAB directly interacts with LBH in NPC cells. This LBH-related pathway we revealed provides a novel therapeutic target for nasopharyngeal carcinoma research. • Focused on cellular metastasis during the recurrence of nasopharyngeal carcinoma. • Epithelial-mesenchymal transition was observed in multiple nasopharyngeal carcinoma cell lines. • Limb-bud and heart gene was studied in nasopharyngeal carcinoma research, including both in vitro and in vivo data. • Fluorescence resonance energy transfer analysis was applied to evaluate protein-protein interaction. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is one of the most common malignant tumors worldwide and tends to have drug resistance. Delicaflavone (DLF), a novel anticancer agent of biflavonoid from Selaginella doederleinii Hieron , showed strong anti-CRC activities, which has not yet been reported. In this study, we investigated the effects and possible anti-CRC mechanism of DLF in vitro and in vivo. It was shown that DLF significantly inhibited the cells viability and induced G2/M phase arrest, apoptosis, the loss of mitochondrial membrane potential (Δψm), generation of ROS and increase of intracellular Ca2+ in HT29 and HCT116 cells by MTT assay, TEM, flow cytometry and inverted fluorescence microscope. Western blot and qPCR assays results further confirmed DLF induced caspase-dependent apoptosis and inhibited PI3K/AKT/mTOR and Ras/MEK/Erk signaling pathways in CRC cells. Meanwhile, DLF significantly suppressed the tumor growth via activation of Caspase-9 and Caspase-3 protein and decrease of ki67 and CD34 protein without apparent side effects in vivo. In summary, these results indicated DLF induced ROS-mediated cell cycle arrest and apoptosis through ER stress and mitochondrial pathway accompanying with the inhibition of PI3K/AKT/mTOR and Ras/MEK/Erk signaling cascade. Thus DLF could be a potential therapeutic agent for CRC. [ABSTRACT FROM AUTHOR]