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Salmonellae, gram-negative bacteria, are facultative intracellular pathogens that cause a number of diseases in animals and humans. The poor penetration ability of antimicrobial agents limits their use in the treatment of intracellular bacterial infections. In this study, the cell-penetrating peptides (CPPs) bLFcin 6 and Tat 11 were separately conjugated to the antimicrobial peptide N2, and the antibacterial activity and pharmacodynamics of the CPPs-N2 conjugates were first evaluated against Salmonellae typhimurium in vitro and in macrophage cells. The cytotoxicity, cellular uptake and mechanism of cellular internalization of the CPPs-N2 conjugates were also examined in RAW264.7 cells. Similar to N2, CPPs-N2 have two reverse β-sheets and three loops. The minimal inhibitory concentration (MIC) of CPPs-N2 was approximately 2 μM, which was higher than that of N2 (0.8 μM). The dose-time curves and cytotoxicity assay showed that both peptide conjugates were more effective than N2 alone at concentrations ranging from 0.25 to 1 × MIC, and they exhibited low cytotoxicity (9.78%–13.54%) at 100 μM. After 0.5 h incubation, the cell internalization ratio of B6N2 and T11N2 exceeded 28.3% and 93.5%, respectively, which was higher than that of N2. The uptake of B6N2 and T11N2 was reduced by low temperature (82.1%–91.7%), chlorpromazine (35.7%–75.1%), and amiloride (26.0%–52.1%), indicating that macropinocytosis and clathrin-mediated endocytosis may be involved. Approximately 98.85% and 91.35% of bacteria were killed within 3 h by T11N2 and B6N2, respectively, which was higher than the percentage killed by N2 (69.74%). Compared with the bactericidal activity of N2 alone, the bactericidal activity of T11N2 and B6N2 was increased by 53.7%–99.6% and 85.3–85.8%, respectively. Both CPPs-N2 conjugates may be excellent candidates for novel antimicrobial agents to treat infectious diseases caused by intracellular pathogens. [ABSTRACT FROM AUTHOR]

Syndromic craniosynostosis is a group of multiple conditions with high heterogeneity, and many rare syndromes still remain to be characterized. To identify and analyze causative genetic variants in nine unrelated probands mainly manifested as syndromic craniosynostosis, we reviewed the relevant medical information of the patients and performed the whole exome sequencing, further verified with Sanger sequencing and parental background. Bioinformatics analysis was used to evaluate the potential deleterious or benign effect of each genetic variant through evolutionary conservation alignment, multi-lines of computer predication and the allele frequency in population dataset (control and patient). The Standards and guidelines from American College of Medical Genetics and Genomics was used to classify and interpret the pathogenicity for each genetic variant. All the nine probands were found to carry the possibly causative variants, among which three variants including two missense mutations (c.3385C > T in IFT122 gene, c.3581A > G in SMC1A gene) and a frameshift mutation (c.434dupA in TWIST1 gene) have never been reported in patients before. We suggested Cornelia de Lange syndrome caused by SMC1A variant is a neglected syndromic craniosynostosis. Our study not only expanded genotypic and phenotypic spectrum of the rare syndromes, but also confirmed that there existed an underlying genetic mechanism. We emphasized that deliberate selection of both the potential candidates and comprehensive detection methods for genetic analysis is important to increase the genetic diagnosis yield of syndromic craninosynostosis. [ABSTRACT FROM AUTHOR]

Objective. The aim of the present study was to investigate the association between dietary patterns and precocious puberty among Shanghai children. Methods. A cross-sectional study was conducted among Shanghai children by multistage stratified cluster random sampling in June 2014. Diet was assessed using a simplified food frequency questionnaire (FFQ). Height, weight, and Tanner stages of breast development, pubic hair growth, and testicular volume were carefully measured. Exploratory factor analysis was used to identify dietary patterns, and logistic regression analysis was used to assess the association between dietary patterns and precocious puberty. Results. Three distinct dietary patterns, “traditional diet,” “unhealthy diet,” and “protein diet,” were established. Neither the “traditional diet” pattern nor the “protein diet” pattern showed any association with precocious puberty, taking gender, BMI, and adjustment factors into consideration. The “unhealthy diet” pattern was significantly positively associated with precocious puberty in both boys (OR = 1.24, 95% CI = 1.02–1.51) and girls (OR = 1.31, 95% CI = 1.10–1.56). The relationship remained positive only for girls (OR = 1.25, 95% CI = 1.04–1.49) after adjustment for age and BMI but statistically nonsignificant after further adjustment for socioeconomic factors in both boys and girls. Conclusions. Dietary patterns were found to be related to precocious puberty among Shanghai children. [ABSTRACT FROM AUTHOR]

Optical vortex attracts many researchers for its interesting properties and promising applications. Focusing properties of the linearly polarized Lorentz-Gaussian beam with azimuthally-variant phase filters were investigated by vector diffraction theory. Results show that the focal intensity distributions can be altered considerably by phase variation parameter of phase filter and the beam parameters, changing phase variation parameter of the phase mask, focal shift, focal switch and focal split may occur, simultaneously, the focal shift direction may change. Moreover, altering phase variation parameter will change the energy distributions of maximum intensity peak and other small intensity peaks. And novel focal patterns also evolve considerably, such as from only one peak to multiple peaks. The focal pattern evolution principle is different with different beam parameter. The tunable focal shift can be used to construct controllable optical tweezers. In practice, azimuthally-variant phase filters can be implemented through phase-only spatial light modulator, which can conveniently alter the wavefront phase distribution of the incident laser beam in the control of computer. [ABSTRACT FROM AUTHOR]

The accumulation and spread of antibiotic resistance bacteria (ARB) in the environment may accelerate the formation of superbugs and seriously threaten the health of all living beings. The timeliness and accurate diagnosing of antibiotic resistance is essential to controlling the propagation of superbugs in the environment and formulating effective public health management programs. Herein, we developed a speedy, sensitive, accurate, and user-friendly colorimetric assay for antibiotic resistance, via a synergistic combination of the peroxidase-like property of the Au–Fe 3 O 4 nanozyme and the specific gene identification capability of the CRISPR-Cas12a. Once the CRISPR-Cas12a system recognizes a target resistance gene, it activates its trans -cleavage activity and subsequently releases the Au–Fe 3 O 4 nanozymes, which oxidizes the 3,3,5,5-tetramethylbenzidine (TMB) with color change from transparent to blue. The diagnosing signals could be captured and analyzed by a smartphone. This method detected kanamycin-resistance genes, ampicillin-resistance genes, and chloramphenicol-resistance genes by simple operation steps with high sensitivity (<0.1 CFU μL−1) and speediness (<1 h). This approach may prove easy for the accurate and sensitive diagnosis of the ARGs or ARB in the field, thus surveilling and controlling the microbial water quality flexibly and efficiently. [Display omitted] • Nanozyme as a novel visual signal reporter for the CRISPR detection system. • Smartphone-based diagnosing makes field detection simple to operate and fast. • CRISPR-nanozyme assay as a rapid and accurate environmental surveillance tool. [ABSTRACT FROM AUTHOR]

Bovine lactoferricin (LfcinB) has potent antibacterial, antifungal and antiparasitic activities but is also hemolytic. Our objective was to identify LfcinB17-31 derivatives with reduced hemolysis and improved antimicrobial activity via substituting Cys3, Arg4, Gln7, Met10, and Gly14 with more hydrophobic residues. Two peptides, Lfcin4 and Lfcin5, showed higher activity against Staphylococcus aureus and Salmonella enteritidis and lower hemolytic activity than the parent peptide LfcinB17-31. These peptides permeabilized the outer and inner membranes of S. enteritidis; however, Lfcin5 did not permeabilize the inner membrane of S. aureus. Gel retardation and circular dichroism spectra showed that Lfcin4 and Lfcin5 bound to bacterial genomic DNA. Lfcin4 inhibited DNA, RNA and protein synthesis. Both peptides induced the peeling of membranes and the lysis of S. enteritidis. At doses of 10 and 15 mg/kg, Lfcin4 and Lfcin5 reduced the bacterial counts in infected thigh muscles by 0.03-0.10 and 0.05-0.63 log10 CFU/g of tissue, respectively, within 10 h. Lfcin4 and Lfcin5 enhanced the survival rate of endotoxemic mice; reduced serum IL-6, IL-1β and TNF-α levels; and protected mice from lipopolysaccharide-induced lung injury. These data suggest that Lfcin4 and Lfcin5 may be antimicrobial and anti-endotoxin peptides that could serve as the basis for the development of dual-function agents. [ABSTRACT FROM AUTHOR]

Background Pathogenic variants of ACAN have been reported to cause spondyloepiphyseal dysplasia Kimberley type, spondyloepimetaphyseal dysplasia, familial osteochondritis dissecans and idiopathic short stature with normal to advanced bone age. A recent international cohort study significantly expanded the ACAN mutation spectrum, further delineated the heterogeneous clinical characteristics of ACAN mutation patients. The prevalence of ACAN mutation in short stature patients is yet unknown. Methods Here we set to assess the frequency of ACAN variants among a cohort of 218 Chinese children with non-syndromic short stature. Results We identified three novel truncating variants at the 5′ end of ACAN gene. All these pathogenic variants co-segregate with severe short stature phenotype in families. In addition, none of the probands showed significant advanced bone age. All affected individuals showed no signs of significant dysmorphic features or skeletal abnormities. The prevalence of ACAN defect in this cohort is estimated to be 1.4% (3/218). It is higher among families with parents also affected with severe short stature, up to 7.0% (3/43) if parental height is < 2.5 SD or 16.7% (3/18) if parental height is < 3.0 SD. Conclusion Our data suggest that ACAN mutation is a relative common cause of familial severe short stature. [ABSTRACT FROM AUTHOR]

Mutations in KAT6B gene are responsible for Say–Barber–Biesecker–Young–Simpson syndrome (SBBYSS) and genitopatellar syndrome (GPS), with most mutations occurring in exon 18. A 4-year-old Chinese boy presented with short stature but no other clinical features of SBBYSS or GPS had a de novo novel nonsense pathogenic mutation in exon 14 of the KAT6B gene at position c.2636T>A (p.Leu879X). The correlation analysis of genotype–phenotype indicated distinctive clinical features (short stature, growth hormone deficiency, and delayed bone age) compared with the classical mutations of KAT6B gene. To the best of our knowledge, this is the first report of KAT6B gene mutation in any Chinese individual. Thiswork expands the mutant phenotypic spectrum of the KAT6B gene. [ABSTRACT FROM AUTHOR]

A Faraday isolator based on magnetically biased graphene is proposed and analyzed. The isolator consists of two rectangular waveguides, two circular-rectangular transitions and a circular cylindrical waveguide loaded multiple graphene sheets, with a static magnetic field which parallel to the signal transmission direction biased. The principles of the isolator have been investigated using the mode-matching technique. The isolation is calculated by the signal flow diagram combined with the transmission matrix method. The theoretical results show that its isolation is more than 30 dB in the working frequency (1.8 GHz–2.2 GHz). The isolation can be tuned by varying the chemical potential or varying the magnetic field. The novel isolator has a simple structure and a wider bandwidth compared to the conventional Faraday isolator. Simultaneously the transmission direction of the isolator can be changed easily by changing the direction of the magnetic field. [ABSTRACT FROM AUTHOR]

Exome sequencing has become a formidable tool for identifying potential de novo variants in causative genes of human diseases, such as neurodegenerative disorders. This article describes a 16-month-old girl with spinal muscular atrophy with lower extremity predominance and a 13-month-old girl with malformations of cortical development. Exome sequencing identified a novel de novo heterozygous missense mutation c.3395G>A (p.Gly1132Glu) and a previously reported de novo heterozygous missense mutation c.10151G>A (p.Arg3384Gln) in the DYNC1H1 gene. Bioinformatics predictions for c.3395G>A and c.10151G>A indicated pathogenicity of the mutations. DYNC1H1 is a pivotal component of cytoplasmic dynein complex, which is a microtubule-related motor involved in retrograde transport. Previous studies indicated that mutant dynein showed decreased run-length of the motor proteins and diminished retrograde transport, which were clearly associated with neuronal death and neurologic diseases. The present findings expand the mutational spectrum of the DYNC1H1 gene, reemphasizing the significance of the DYNC1H1 protein in the functioning of neurons. [ABSTRACT FROM AUTHOR]

NZ2114 is a promising candidate for therapeutic application owing to its potent activity to Staphylococcus aureus. Our objective was to identify NZ2114 derivatives with improved activity through substitution of His16 and His18 with residues Arginine and Lysine. Eight mutants were designed and expressed in Pichia pastoris X-33 via pPICZαA. Five of them exhibited strong antimicrobial activity against S. aureus at low minimal inhibitory concentrations (MICs) of 0.057-0.454 μM. Among them, H1, H2, and H3 showed ideal pharmacodynamic effects on methicillin-resistant S. aureus ATCC43300. The total protein level of H1, H2, and H3 reached 1.70, 1.77 and 1.54 g/l at 120 h of induction in the 5-l fermenter, respectively. They killed over 99.9% of pathogens within 1.5 h at 2× and 4× MIC. The post antibiotic effect of H1, H2 and H3 to S. aureus ATCC43300 was 2.94, 1.75 and 1.55 h at 2× MIC, which was similar with their original peptide NZ2114 (1.43 h) and vancomycin (1.72 h). The fractional inhibitory concentration index (FICI) indicated indifferent effects between H1, H2, H3 and vancomycin, ampicillin, rifampicin. Additionally, they had low hemolysis and high stability in different environments (temperature, pH, proteases, and saline ions). All results indicate that H1, H2, and H3 can be produced in large-scale and have potential as therapeutic drugs against MRSA. [ABSTRACT FROM AUTHOR]

NZ2114 is a promising candidate for therapeutic application owing to potent activity to gram-positive bacterium such as Streptococcus pneumoniae and Staphylococcus aureus. This work is the first report to describe the in vitro and in vivo antibacterial characteristics of NZ2114 against Streptococcus suis. It exhibited strong antimicrobial activity against S. suis type 2 strains CVCC 606, CVCC 3309, and CVCC 3928 at a low minimal inhibitory concentration (MIC) of 0.03-0.06 μM. The NZ2114 killed over 99.9% of tested S. suis CVCC 606 in Mueller-Hinton medium within 4 h when treated with 4 × MIC. It caused only less than 0.25% hemolytic activity in the concentration of 256 μg/ml. Additionally, NZ2114 exhibited potent in vivo activity to S. suis. All mice were survival when the dosage was low to 0.2 mg/kg. Over 99% of S. suis cells were killed within 4 h in blood, lung, liver and spleen with dosage of 10, 20, and 40 mg/kg in mice peritonitis models and no pathogen were detected after 24 h of treatment. Further, no pathological phenomenon in lung and low level of inflammatory cytokines in blood were detected. These results indicate that NZ2114 has the potential to be a new antimicrobial agent candidate for the clinical treatment of infection caused by S. suis type 2. [ABSTRACT FROM AUTHOR]

Tubular interstitial extracellular matrix accumulation, which plays a key role in the pathogenesis and progression of diabetic kidney disease (DKD), is believed to be mediated by activation of PI3K/Akt signal pathway. However, it is still not clear whether SH2 domain-containing inositol 5'-phosphatase (SHIP), known as a negative regulator of PI3K/Akt pathway is also involved in extracellular matrix metabolism of diabetic kidney. In the present study, decreased SHIP and increased phospho-Akt (Ser 473, Thr 308) were found in renal tubular cells of diabetic mice accompanied by overexpression of connective tissue growth factor (CTGF) and extracellular matrix deposition versus normal mice. Again, high glucose attenuated SHIP expression in a time-dependent manner, concomitant with activation of PI3K/Akt signaling and extracellular matrix production in human renal proximal tubular epithelial cells (HK2) cultured in vitro , which was significantly prevented by transfection of M90-SHIP vector. Furthermore, in vivo delivery of rAd-INPP5D vector (SHIP expression vector) via intraperitoneal injection in diabetic mice increased SHIP expression by 3.36 times followed by 65.26%, 70.38% and 46.71% decreases of phospho-Akt (Ser 473), phospho-Akt (Thr 308) and CTGF expression versus diabetic mice receiving rAd-EGFP vector. Meanwhile, increased renal extracellular matrix accumulation of diabetic mice was also inhibited with intraperitoneal injection of rAd-INPP5D vector. These above data suggested that overexpression of SHIP might be a potent method to lessen renal extracellular matrix accumulation via inactivation of PI3K/Akt pathway and suppression of CTGF expression in DKD. [ABSTRACT FROM AUTHOR]

Abstract COG6-CDG is a rare autosomal recessive disease of congenital disorders of glycosylation (CDG) caused by deficiency of subunit 6 of the conserved oligomeric Golgi complex (COG6), which is characterized by growth retardation, developmental disability, microcephaly, liver and gastrointestinal disease, recurrent infections and hypohidrosis/hyperthermia. Only eight mutations causing COG6 deficiencies have been described since the first report in 2010. Here, we report the first Chinese patient with COG6-CDG. Utilizing targeted next generation sequencing and Sanger sequencing, we detected compound heterozygous variants (c.1A > G, p.? and c.388C > T, p.(Gln 130*)) of the COG6 gene, both of which were pathogenic. Our study therefore extended the genotype-phenotype relationship of the COG6 gene. [ABSTRACT FROM AUTHOR]

A correction to the article "Design, expression, and characterization of a novel targeted plectasin against methicillin-resistant Staphylococcus aureus" by Ruoyu Mao and colleagues that was published in the March 2013 issue is presented.

Background: Epidemiological studies have investigated the role of choline acetyltransferase (ChAT) in Alzheimer’s disease (AD). ChAT gene polymorphisms (SNPs rs868750G/A, rs1880676G/A, rs2177369G/A, and rs3810950G/A) may be associated with the risk of AD. In this meta-analysis, we determined the relationship between the four polymorphisms and the risk of AD. Methods: We searched MEDLINE, EMBASE, and HuGEnet databases for studies linking the four polymorphisms with AD risk. We included 16 articles in our meta-analysis to assess the association between the four polymorphisms and susceptibility to AD by calculating the pooled odds ratios (ORs) and 95% confidence intervals (CIs). Results: The combined results showed no significant association with rs1880676G/A and rs2177369G/A polymorphisms. The risk of AD (GG+GA versus AA: OR = 0.01, 95%CI = 0.01–0.02, P < 0.05; GG versus GA+AA: OR = 0.85, 95%CI = 0.72–1.00, P = 0.05; GA versus AA: OR = 0.60, 95% CI = 0.37–0.98, P = 0.04) with rs868750G/A polymorphism, or the association of rs3810950G/A polymorphism with AD risk in the overall population (GA versus AA: OR = 0.64, 95% CI = 0.44–0.93, P = 0.02; GG+GA versus AA: OR = 0.62, 95% CI = 0.39–0.97, P = 0.04) or Asian group (GA versus AA: OR = 0.50, 95% CI = 0.32–0.76, P = 0.001, and GG+GA versus AA: OR = 0.46, 95% CI = 0.30–0.09, P = 0.0002) was demonstrated. Conclusions: Our meta-analysis suggested that rs1880670G/A, and rs2177369 G/A polymorphisms were not risk factors for AD. However, rs3810950G/A, or rs868750G/A genetic polymorphism was a genetic risk factor for the development of AD. The rs3810950G/A polymorphism had a negative effect on the risk of AD for GA or GG+GA genotypes compared with AA in the overall population or Asians. [ABSTRACT FROM AUTHOR]

Background: Stroke is a multifactorial disease in which genetic factors play an important role. Previous studies associated angiotensin converting enzyme (ACE) (insertion/deletion, I/D) gene polymorphism with ischemic stroke risk in Caucasian individuals reported conflicting results. The purpose of this study was to evaluate the association between ACE (I/D) gene polymorphism and ischemic stroke risk by a meta-analysis.Methods:The related studies were searched in MEDLINE, EMBASE and HuGEnet databases. The odds ratios (ORs) and corresponding 95% confidence intervals (CIs) for ischemic stroke risk associated with this polymorphism were estimated using fixed-effect or random-effects model. Twenty-two studies (5528/5081 cases/controls) were eligible in our meta-analysis.Results: Overall, statistical associations of the ACE (I/D) polymorphism with ischemic stroke risk were found in dominant model (DD + ID versus II) : OR = 1.21, 95% CI = (1.06,1.38),P= 0.006, recessive model (DD versus ID + II): OR = 1.28, 95% CI = (1.05,1.55),P= 0.01, and homozygote comparison (DD versus II): OR = 1.37, 95% CI = (1.14,1.65),P= 0.001 for Caucasians. When stratifying according to stroke subtypes, there were similarly significant differences for small vessel disease in dominant model (DD + ID versus II) : OR = 1.44, 95% CI = (1.01,2.05),P= 0.04, recessive model (DD versus ID + II): OR = 1.30,95% CI = (1.09,1.55),P= 0.004, and homozygote comparison (DD versus II): OR = 1.44, 95% CI = (1.15,1.80),P= 0.001.Conclusion: This analysis suggests that the ACE (I/D) polymorphism may be a risk factor for ischemic stroke, genotype DD of ACE could increase the risk of ischemic stroke in Caucasians. Subgroup analyses indicate that stroke subtypes may be a genetic risk factor of ischemic stroke, and there might be a greater genetic liability with small vessel disease. [ABSTRACT FROM AUTHOR]

Plectasin, the first fungus-derived defensin from Pseudoplectania nigrella , is considered to be one of the most likely substitutes for antibiotics used to cure Staphyloccocus and Streptococcus infections. Recombinant expression of plectasin in Pichia pastoris X-33 was achieved. In this work, we aimed to develop a multi-copy gene cassette to enhance plectasin expression in P. pastoris . Expression vectors pPICZαA-Pn ( n = 1, 2, 4, or 8) harboring a one, two, four, or eight-copy gene cassette were constructed and confirmed by digestion with Bam HI and Bgl II. Vectors linearized by Bgl II were transformed into P. pastoris by electroporation. The gene copy number of the recombinants was quantified by real-time quantitative PCR. Yields of recombinant plectasin were 296, 409, 516, and 879 mg/L for the 1-, 2-, 4-, and 8-copy transformants, respectively, in a 5-L fermentor, demonstrating that an increase in copy number results in a proportional elevation in the expression level of recombinant plectasin. The purity of plectasin was 90.8%, and the yield was found to be 814 mg/L using ion-exchange chromatography. Multi-copy gene dosage enhances plectasin expression levels and highly reduces its production cost. Additionally, the above results could be helpful in improving the application and understanding of the P. pastoris expression system. [ABSTRACT FROM AUTHOR]

A novel antimicrobial peptide MP1106 was designed based on the parental peptide plectasin with four mutational sites and a high level of expression in Pichia pastoris X-33 via the pPICZαA plasmid was achieved. The concentration of total secreted protein in the fermented supernatant was 2.134 g/l (29 °C), and the concentration of recombinant MP1106 (rMP1106) reached 1,808 mg/l after a 120-h induction in a 5-l fermentor. The rMP1106 was purified using a cation-exchange column, and the yield was 831 mg/l with 94.68 % purity. The sample exhibited a narrow spectrum against some Gram-positive bacteria and strong antimicrobial activity against Staphylococcus aureus at low minimal inhibitory concentrations (MICs) of 0.014, 1.8, 0.45, and 0.91 μM to S. aureus strains ATCC 25923, 29213, 6538, and 43300, respectively. Meanwhile, rMP1106 showed potent activity (0.03-1.8 μM) against 20 clinical isolates of methicillin-resistant S. aureus (MRSA). In addition, rMP1106 exhibited a broad range of thermostability from 20 to 100 °C. The higher antimicrobial activity of rMP1106 was maintained in neutral and alkaline environments (pH 6, 8, and 10), and its activity was slightly reduced in acidic environments (pH 2 and 4). The rMP1106 was resistant to the digestion of pepsin, snailase, and proteinase K and was sensitive to trypsin. It exhibited hemolytic activity of only 1.16 % at a concentration of 512 μg/ml and remained stable in human serum at 37 °C for 24 h. Furthermore, the activity of rMP1106 was minorly affected by 10 mM dithiothreitol and 20 % dimethylsulfoxide. Our results indicate that MP1106 can be produced on a large scale and has potential as a therapeutic drug against S. aureus. [ABSTRACT FROM AUTHOR]

For preventing premature drug release in neutral environment and avoiding them being trapped into the endosomal/lysosomal system, we developed a novel iron silicate@liposome hybrid (ILH) formulation, which can be used as a carrier to transport doxorubicin (DOX) in a pH-sensitive manner and to escape from endosomal/lysosomal trapping through “proton-sponge” effect. The high intensity of photoacoustic signal from in vitro photoacoustic imaging (PAI) experiments suggests that it is a promising candidate for PAI agent, providing the potential for simultaneously bioimaging and cancer-targeting drug delivery. Cytotoxicity of our formulation toward tumor cells was remarkably higher than free DOX (48.4±7.7% and 26.2±8.4%, P<0.001). Confocal laser scanning microscopy experiments showed the enhanced transportation and enrichment process of DOX in QSG-7703 cells. Taking together, we developed an easy approach to construct a multifunctional anticancer drug delivery/imaging system with a potency as a PAI agent. The strategy of combining drug carrier and imaging agent is an emerging platform for further construction of nanoparticle and may play a significant role in cancer therapy and diagnosis. [ABSTRACT FROM AUTHOR]

The programmed death-1 (PD-1) molecule is mainly expressed on functionally 'exhausted' CD8 T cells, dampening the host antitumor immune response. We evaluated the ratio between effective and regulatory T cells (Tregs) and PD-1 expression as a prognostic factor for operable breast cancer patients. A series of 218 newly diagnosed invasive breast cancer patients who had undergone primary surgery at Ruijin Hospital were identified. The influence of CD8 cytotoxic T lymphocytes, FOXP3 (Treg cell marker), and PD-1 immune cell counts on prognosis was analyzed utilizing immunohistochemistry. Both PD-1 immune cells and FOXP3 Tregs counts were significantly associated with unfavorable prognostic factors. In bivariate, but not multivariate analysis, high tumor infiltrating PD-1 cell counts correlated with significantly shorter patient survival. Our results suggest a prognostic value of the PD-1 immune cell population in such breast cancer patients. Targeting the PD-1 pathway may be a feasible approach to treating patients with breast cancer. [ABSTRACT FROM AUTHOR]

Highlights: [•] First report of cloning of a novel AP gene from Saccharomyces boulardii. [•] Heterologous expression of high-activity rAP in Pichia pastoris X-33. [•] rAP possess favorable enzymatic characters. [•] rAP possess the function of dephosphorylation of LPS. [•] rAP alleviate inflammation responses caused by LPS. [Copyright &y& Elsevier]

NZ2114, a new variant of plectasin, was overexpressed in Pichia pastoris X-33 via pPICZαA for the first time. The total secreted protein of fermentation supernatant reached 2,390 mg/l (29 °C) and 2,310 mg/l (25 °C), and the recombinant NZ2114 (rNZ2114) reached 860 mg/l (29 °C) and 1,309 mg/l (25 °C) at 96 h induction in a 5-l fermentor, respectively.The rNZ2114 was purified by cation exchange chromatography, and its yield was 583 mg/l with 94.8 % purity. The minimal inhibitory concentration (MIC) of rNZ2114 to four ATCC strains of Staphyloccocus aureus was evaluated from 0.028 to 0.90 μM. Meanwhile, it showed potent activity (0.11-0.90 μM) to 20 clinical isolates of MRSA. The rNZ2114 killed over 99.9 % of tested S. aureus (ATCC 25923 and ATCC 43300) in Mueller-Hinton medium within 6 h when treated with 4 × MIC. The postantibiotic effect of rNZ2114 to S. aureus ATCC 25923 and ATCC 43300 was 18.6-45.6 and 1.7-3.5 h under 1×, 2×, and 4× MIC, respectively. The fractional inhibitory concentration index (FICI) indicated a synergistic effect between rNZ2114 and kanamycin, streptomycin, and vancomycin against S. aureus ATCC 25923 (FICI = 0.125), and additivity between rNZ2114 and ampicillin, spectinomycin (FICI = 0.625), respectively. To S. aureus ATCC 43300 [methicillin-resistant S. aureus (MRSA)], rNZ2114 showed a synergistic effect (FICI = 0.125-0.3125) with kanamycin, ampicillin, streptomycin, and vancomycin, and antagonism with spectinomycin (FICI = 8.0625). The rNZ2114 caused only less than 0.1 % hemolytic activity in the concentration of 128 μg/ml, and showed a good thermostability from 20 to 80 °C. In addition, it exhibited the highest activity at pH 8.0. These results suggested that large-scale production of NZ2114 is feasible using the P. pastoris expression system, and it could be a new potential antimicrobial agent for the prevention and treatment of S. aureus especially for MRSA infections. [ABSTRACT FROM AUTHOR]

Abstract: The alpha subunit of β-conglycinin is one of the main allergens found in soybeans. For the preparation of a specific monoclonal antibody (mAb) against the α subunit, potential epitopes were predicted using Protean and evaluated by Wu''s Antigenic Index. The specific epitope 85EQDERQFPFPR95 was synthesized, and then conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) for use as an immunogen and the plate-coating antigen, respectively. The resulting mAb, termed mAb-60K, was characterized as belonging to the IgG1 isotype and containing the κ light chain; it exhibited high specificity for the α subunit and did not cross react with the α′ and β subunits of β-conglycinin or other proteins found within soybeans. A competitive enzyme-linked immunosorbent assay (cELISA) with high accuracy and reproducibility was developed based on mAb-60K and the plate-bound peptide. Co-incubation with the full-length α subunit showed a 50% mAb binding inhibition concentration (IC50) value of 4.42ng/mL, with a linear inhibition curve observed α subunit concentrations between 0.65 and 29.84ng/mL. The newly developed mAb-60K and the companion cELISA could provide a valuable tool for determining sensitivity towards the α subunit of soybean β-conglycinin and for future studies on food allergies resulting from this protein. [Copyright &y& Elsevier]

Aims: To determine whether administration of long-acting polyethylene glycol moiety covalently linked to recombinant human growth hormone (PEG-rhGH) can be efficacious in the treatment of small for gestational age (SGA) rats and to characterize its effects on metabolic parameters. Methods: 20 pregnant rats were randomly divided into undernourished and standard nourished groups. SGA or appropriate for gestational age (AGA) offspring from each were then randomly assigned to receive either PEG-rhGH or normal saline (NS). Once-weekly subcutaneous injections of PEG-rhGH (2 μg/g/week) were administered starting at 21-42 days. Glycometabolism, blood pressure (BP), hormone and biochemical levels were analyzed at 21, 42 and 70 days. Results: SGA rats at 21 days were lighter and shorter than AGAs (34.77 ± 2.11 vs. 48.83 ± 1.78 g, 10.42 ± 0.22 vs. 12.99 ± 0.17 cm, p < 0.05). At 42 days, SGA animals experienced a greater body weight gain. BP was higher in NS-treated SGA than in the AGA group at 70 days (138.16 ± 3.02 vs. 112.26 ± 5.42 mm Hg, p < 0.05). Meanwhile, NS-treated SGA exhibited higher glucose levels at 60 and 90 min than the AGA groups. No differences in hormone levels between the SGA and AGA groups were found at the end of PEG-rhGH treatment. Conclusion: These data suggest that PEG-rhGH treatment does not increase the risk of developing metabolic syndrome in adolescent aged rats. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: SHORT syndrome is a rare genetic disease named with the acronyms of short stature, hyper-extensibility of joints, ocular depression, Rieger anomaly and teething delay. It is inherited in an autosomal dominant manner confirmed by the identification of heterozygous mutations in PIK3R1. This study hereby presents a 15-year-old female with intrauterine growth restriction, short stature, teething delay, characteristic facial gestalts who was identified a novel de novo nonsense mutation in PIK3R1. Case presentation: The proband was admitted to our department due to irregular menstrual cycle and hirsutism with short stature, who had a history of intrauterine growth restriction and presented with short stature, teething delay, characteristic facial gestalts, hirsutism, and thyroid disease. Whole-exome sequencing and Sanger sequencing revealed c.1960C > T, a novel de novo nonsense mutation, leading to the termination of protein translation (p. Gln654*). Conclusions: This is the first case report of SHORT syndrome complicated with thyroid disease in China, identifying a novel de novo heterozygous nonsense mutation in PIK3R1 gene (p. Gln654*). The phenotypes are mildly different from other cases previously described in the literature, in which our patient presents with lipoatrophy, facial feature, and first reported thyroid disease. Thyroid disease may be a new clinical symptom of patients with SHORT syndrome. [ABSTRACT FROM AUTHOR]

• Probiotics have different physiological functions on immune and microecology. • There are two types of probiotics resistance to antibiotics: intrinsic and acquired. • Probiotics develop antibiotic resistance by gene mutation or horizontal gene transfer. • Probiotics have potential risks-pathogenicity and antibiotic resistance gene transfer. Probiotics are defined as live microorganisms that, when administered in adequate amounts, confer a health benefit upon the host. At present, probiotics are gaining popularity worldwide and are widely used in food and medicine. Consumption of probiotics is increasing with further in-depth research on the relationship between intestinal flora and host health. Most people pay more attention to the function of probiotics but ignore their potential risks, such as infection and antibiotic resistance transfer to pathogenic microbes. Physiological functions, effects and mechanisms of action of probiotics were covered in this review, as well as the antibiotic resistance phenotypes, mechanisms and genes found in probiotics. Typical cases of antibiotic resistance of probiotics were also highlighted, as well as the potential risks (including pathogenicity, infectivity and excessive immune response) and corresponding strategies (dosage, formulation, and administration route). This timely study provides an avenue for further research, development and application of probiotics. [ABSTRACT FROM AUTHOR]

More than 30% of patients with epilepsy progress to drug-resistant epilepsy, leading to a significant increase in morbidity and mortality of epilepsy. The limitation of epileptic drug to reach the epileptogenic focus is the critical reason, and the blood-brain barrier (BBB) plays a crucial role. Here, we successfully constructed a hepatitis B core (HBc) protein nanocage (NC) with the insertion of brain target TGN peptide for facilitating epileptic drug phenytoin delivery to the brain. Our results demonstrated that this nanocage can specifically and efficiently target the brain tissue by 2.4 fold and increase the antiepileptic efficiency of phenytoin about 100 fold in pilocarpine induced models of epilepsy. Both in vivo mice and in vitro human neural three-dimensional cortical organoids demonstrated high penetration ability. These functions are achieved through the facilitation of brain target peptide TGN rather than disruption of brain blood barrier. In summary, we presented an efficient antiepileptic drug delivery nanocage for the treatment of refractory epilepsy. Moreover, this therapeutic modulation also provides promising strategy for other intractable neurological disease. [ABSTRACT FROM AUTHOR]

• The photoelectric performance of MoS2/WSe2 heterojunctions was studied in a PEC cell. • Heterojunctions were fabricated via liquid-phase exfoliation and vacuum filtration. • The band alignment of the heterojunctions was established with the support of XPS. • The mechanism of the photoelectrochemistry process was revealed. Fabricating a two-dimensional (2D) van der Waals heterostructure is an efficient strategy to improve the photoelectric performance of 2D materials, thus providing a new possibility for the photoanode material design in photoelectrochemistry (PEC) devices. Herein, transition metal dichalcogenide (TMD) MoS 2 /WSe 2 heterojunction photoelectrodes are prepared via liquid-phase exfoliation and vacuum filtration. Linear sweep voltammetry, transient photocurrent, and open circuit potential measurements show that the heterojunction photoelectrodes have enhanced photocurrent intensity and improved photoresponse activity. Moreover, Nyquist impedance plots, Bode phase plots, and Mott-Schottky measurements demonstrate that the heterojunction samples have higher charge transfer rate and longer charge lifetime than MoS 2 and WSe 2. To reveal the mechanisms, the band alignment of MoS 2 /WSe 2 heterojunction samples is established with the support of X-ray photoelectron spectroscopy and first-principles theory calculation. Accordingly, the improved PEC performance of the heterojunction photoanode is ascribed to the built-in electric field between MoS 2 and WSe 2 nanosheets, which promotes the photogenerated e-h pair separation and suppresses their recombination. This work verifies the PEC mechanism of MoS 2 /WSe 2 heterojunction photoelectrodes, which is significant for the design of PEC devices based on TMD heterostructures. [ABSTRACT FROM AUTHOR]

Background: Gain-of-function pathogenic variants of the Erb-B2 receptor tyrosine kinase 3 (ERBB3) gene contribute to the occurrence and development of a variety of human carcinomas through activation of phosphatidylinositol 3-kinase (PI3K)/AKT and extracellular signal-regulated kinase (ERK) signaling. ERBB3 gene homozygous germline variants, whose loss of function may cause autosomal recessive congenital contractural syndrome, were recently identified. This study aims to identify the disease-causing gene in a Chinese pedigree with variable phenotypes involving multiple systems, including developmental delay, postnatal growth retardation, transient lower limb asymmetry, facial malformations, atrioventricular canal malformation, bilateral nystagmus and amblyopia, feeding difficulties, immunodeficiency, anemia, and liver damage, but without congenital contracture.Methods: Trio-whole exome sequencing (WES) was performed to identify the disease-causing gene in a 24-month-old Chinese female patient. The pathogenicity of the identified variants was evaluated using in silico tools and in vitro functional studies.Results: Trio-WES revealed compound heterozygous variants of c.1253 T > C (p.I418T) and c.3182dupA (p.N1061Kfs*16) in the ERBB3 gene. Functional studies showed that p.I418T resulted in normal expression of ERBB3, which was capable of interacting with ERBB2. However, the variant impaired ERBB3 phosphorylation, consequently blocking ERBB2 phosphorylation and AKT and ERK activation. The truncated protein resulting from the c.3182dupA variant also lacked the capacity to activate downstream signaling pathways.Conclusions: We report the first patient with a novel multisystem syndrome disorder without congenital contracture resulting from biallelic loss-of-function variants of ERBB3. [ABSTRACT FROM AUTHOR]

Background: Neurofibromatosis 1 (NF1) is one of the most common dominantly inherited genetic disorders worldwide, with an age-dependent phenotypic expression. Exploring the mutational spectrum and clinical presentation of NF1 patients at different ages from a diverse population will aid the understanding of genotype–phenotype correlations. Methods: In this study, 95 Chinese children with clinical suspicion of NF1 mainly due to the presence of multiple café-au-lait macules (CALMs) were subjected to medical exome-sequencing analysis and Sanger confirmation of pathogenic variants. Clinical presentations were evaluated regarding dermatological, ocular, neurological, and behavioral features. Results: Pathogenic or likely pathogenic NF1 variants were detected in 71.6% (68/95) of patients; 20 pathogenic variants were not previously reported, indicating that Chinese NF1 patients are still understudied. Parental Sanger sequencing confirmation revealed 77.9% of de novo variants, a percentage that was much higher than expected. The presence of a higher number of NF1-related features at young ages was correlated with positive diagnostic findings. In addition to CALMs, neurological and behavioral features had a high expression among Chinese NF1 children. We attempted to correlate short stature with the locations of the pathogenic variants across the NF1 gene. It is interesting to notice that variants detected in the C-terminal region of the NF1 gene were less likely to be associated with short stature among the NF1 patients, whereas variants at the N-terminal were highly penetrant for the short stature phenotype. Conclusion: Novel NF1 pathogenic variants are yet to be uncovered in under-studied NF1 patient populations; their identification will help to reveal novel genotype–phenotype correlations. [ABSTRACT FROM AUTHOR]

Background: Wiedemann-Steiner syndrome (WDSTS) is a rare genetic disorder characterized by facial gestalt, neurodevelopmental delay, skeletal anomalies and growth retardation, which is caused by variation of KMT2A gene. To date, only 2 Chinese WDSTS patients have been reported. Here, we report the phenotypes and KMT2A gene variations in 14 unrelated Chinese WDSTS patients and investigate the phenotypic differences between the Chinese and French cohorts.Methods: Next generation sequencing was performed for each patient, and the variants in the KMT2A gene were validated by Sanger sequencing. The phenotypes of 16 Chinese WDSTS patients were summarized and compared to 33 French patients.Results: Genetic sequencing identified 13 deleterious de novo KMT2A variants in 14 patients, including 10 truncating, 2 missenses and 1 splicing variants. Of the 13 variants, 11 are novel and two have been reported previously. One of the patients is mosaic in the KMT2A gene. The variation spectra and phenotypic profiles of the Chinese WDSTS patients showed no difference with patients of other ethnicities; however, differ in the frequencies of several clinical features. We demonstrated that variations in the KMT2A gene can lead to both advanced and delayed bone age. We identified 6 novel phenotypes, which include microcephaly, deep palmar crease, external ear deformity, carpal epiphyseal growth retardation, dyslipidemia, and glossoptosis. In addition, patients harbored missense variants in the CXXC zinc finger domain of KMT2A showed more severe neurophenotypes.Conclusion: Our study consists of the largest cohort of Chinese WDSTS patients that continues to expand the WDSTS phenotypic and variation spectrum. Our results support the notion that the CXXC zinc finger domain of KMT2A gene is a hotspot for missense variants associated with more severe neurophenotypes. [ABSTRACT FROM AUTHOR]