Targeted polyethylenimine/(p53 plasmid) nanocomplexes for potential antitumor applications.

The development of the tumor-targeting ability of nanocarriers is of paramount importance for gene delivery into tumor lesions as well as to avoid biotoxicity. Here we report the synthesis of the polyethyleneimine-fluorescein isothiocyanate-folic acid (PEI-FITC-FA) polymer, which could condense the tumor suppressor pp53 to form nanocomplexes. These targeted nanocomplexes exhibited favorable physical properties including a small size of <100 nm, exploiting the enhanced permeability and retention effect and tumor-targeting ability by binding to the overexpressed FA receptors on tumor cell surfaces. In addition, once the nanocomplexes are accumulating in the tumor tissue, the target functional ligand, FA, can selectively recognize the over-expressed FA receptor and subsequently remain on the tumor cell surface, which can significantly promote the tumor cell uptake because of the time- and concentration-dependent internalization caused by the enhanced interaction between nanocomplex and tumor cell. Our results indicated that PEI-FITC-FA/pp53 nanocomplexes could be efficiently delivered into tumor cells, and subsequently induce tumor cell apoptosis. Thus, the targeted cationic polymer PEI-FITC-FA could be used as an advanced nanocarrier for gene delivery.

Dynamic Dispersal of Surface Layer Biofilm Induced by Nanosized TiO2 Based on Surface Plasmon Resonance and Waveguide.

Pollutant degradation is present mainly in the surface layer of biofilms, and the surface layer is the most vulnerable to impairment by toxic pollutants. In this work, the effects of nanosized TiO2 (n-TiO2) on the average thicknesses of Bacillus subtilis biofilm and on bacterial attachment on different surfaces were investigated. The binding mechanism of n-TiO2 to the cell surface was also probed. The results revealed that n-TiO2 caused biofilm dispersal and the thicknesses decreased by 2.0 to 2.6 µm after several hours of exposure. The attachment abilities of bacteria with extracellular polymeric substances (EPS) on hydrophilic surfaces were significantly reduced by 31% and 81% under 10 and 100 mg/liter of n-TiO2, respectively, whereas those of bacteria without EPS were significantly reduced by 43% and 87%, respectively. The attachment abilities of bacteria with and without EPS on hydrophobic surfaces were significantly reduced by 50% and 56%, respectively, under 100 mg/liter of n-TiO2 The results demonstrated that biofilm dispersal can be attributed to the changes in the cell surface structure and the reduction of microbial attachment ability.IMPORTANCE Nanoparticles can penetrate into the outer layer of biofilm in a relatively short period and can bind onto EPS and bacterial surfaces. The current work probed the effects of nanosized TiO2 (n-TiO2) on biofilm thickness, bacterial migration, and surface properties of the cell in the early stage using the surface plasmon resonance waveguide mode. The results demonstrated that n-TiO2 decreased the adhesive ability of both cell and EPS and induced bacterial migration and biofilm detachment in several hours. The decreased adhesive ability of microbes and EPS worked against microbial aggregation, reducing the effluent quality in the biological wastewater treatment process.

Acid-Activated Melittin for Targeted and Safe Antitumor Therapy.

Melittin (MLT), as a natural active biomolecule, can penetrate the tumor cell membrane to play a role in cancer treatment and will attract more attention in future development of antitumor drugs. The main component of natural bee venom MLT was modified by introducing a pH-sensitive amide bond between the 2,3-dimethyl maleimide (DMMA) and the lysine (Lys) of MLT (MLT-DMMA). MLT and its corresponding modified peptide MLT-DMMA were used for antitumor and biocompatibility validation. The biomaterial characteristics were tested by MALDI-TOF MS, 1H NMR, IUPAC and HPLC, cell viability, hemolytic and animal experiment safety evaluation. Compared with the primary melittin, the modified peptide showed decreased surface charge and low cytotoxicity in physiological conditions. Moreover, cell assays confirmed the acid-activated conversion of amide bond resulting in adequate safety during delivery and timely antitumor activity in tumor lesions. Thus, MLT-DMMA provided a feasible platform to improve the targeted and safe antitumor applications.

Analysis of bioprinting strategies for skin diseases and injuries through structural and temporal dynamics: historical perspectives, research hotspots, and emerging trends.

This study endeavors to investigate the progression, research focal points, and budding trends in the realm of skin bioprinting over the past decade from a structural and temporal dynamics standpoint. Scholarly articles on skin bioprinting were obtained from WoSCC. A series of bibliometric tools comprising R software, CiteSpace, HistCite, and an alluvial generator were employed to discern historical characteristics, evolution of active topics, and upcoming tendencies in the area of skin bioprinting. Over the past decade, there has been a consistent rise in research interest in skin bioprinting, accompanied by an extensive array of meaningful scientific collaborations. Concurrently, diverse dynamic topics have emerged during various periods, as substantiated by an aggregate of 22 disciplines, 74 keywords, and 187 references demonstrating citation bursts. Four burgeoning research subfields were discerned through keyword clustering-namely, #3 'in situbioprinting', #6 'vascular', #7 'xanthan gum', and #8 'collagen hydrogels'. The keyword alluvial map reveals that Module 1, including 'transplantation' etc, has primarily dominated the research module over the previous decade, maintaining enduring relevance despite annual shifts in keyword focus. Additionally, we mapped out the top six key modules from 2023 being 'silk fibroin nanofiber', 'system', 'ionic liquid', 'mechanism', and 'foot ulcer'. Three recent research subdivisions were identified via timeline visualization of references, particularly Clusters #0 'wound healing', #4 'situ mineralization', and #5 '3D bioprinter'. Insights derived from bibliometric analyses illustrate present conditions and trends in skin bioprinting research, potentially aiding researchers in pinpointing central themes and pioneering novel investigative approaches in this field.

[Effect of horizontal rotary culture on zebrafish vascular development].

With the development of space life science, a study on the influence of microgravity on organism has been an increasingly concerned topic. Lots of studies indicate that microgravity plays an important role in the early development of embryos. The vascular system as the first-function system of embryos provides an interesting topic for many researchers. However, those studies were mostly carried out in vitro by rotary cell culture system (RCCS), while few experiments were done in vivo. Using zebrafish as a model, this research investigated the effects of horizontal rotary culture on the vascular development in vivo. Zebrafish embryos at 24 hpf (hour post-fertilization) were selected and divided into two groups. One group was cultured by the shaker, and the other was cultured normally as the control. After 12 h, all the embryos were collected and detected. The phenotype of zebrafish was observed by stereo microscope. Then, the expression of vascular specific expression factor, flk1, flt4, and ephrinB2 was compared by RT-PCR, qPCR, and in situ hybridization, respectively. Cell apoptosis and proliferation in situ were observed using TUNEL assay and bromodeoxyuridine incorporation. The results demonstrated that horizontal rotary culture at 90 r/min decreased the hatching of embryos (10.3±0.41 vs. 0.0, P<0.05), accelerate the heart rate (223.5±2.32 vs. 185.0±3.23, P<0.05) and increased the content of melanin in zebrafish significantly. At the same time, we found some differences in the vascular system of zebrafish after horizontal rotary culture which caused a down regulation of flk1, flt4, and ephrinB2. On the other hand, horizontal rotary culture accelerated the apoptosis of cells in zebrafish, but showed no significance in proliferation. In conclusion, horizontal rotary culture has a significant influence on the vascular development in zebrafish.

High-level production of a cold-active B-mannanase from Bacillus subtilis BS5 and its molecular cloning and expression.

Mannanases can be useful in the food, feed, pulp and paper industries. In this research a Bacillus subtilis strain (named Bs5) which produced high-level beta-mannanase was isolated. Maximum level of beta-mannanase (1231.41 U/ml) was reached when Bacillus subtilis Bs5 was grown on konjac powder as the carbon source for nine hours at 32 degrees C. The beta-mannanase was a typical cold-active enzyme and its optimal temperature of 35 degrees C was the lowest among those of the known mannanases from bacteria. In addition, the optimal pH was 5.0 and much wide pH range from 3.0-8.0 was also observed in the beta-mannanase. These properties make the beta-mannanase more attractive for biotechnological applications. The DNA sequence coding the beta-mannanase was cloned and the open reading frame consisted of 1089 bp encoding 362 amino acids. A phylogenetic tree of the beta-mannanase based on the similarity of amino acid sequences revealed that the beta-mannanase formed a cluster with the beta-mannanases of Bacillus subtilis, which was separated from the mannanases of fungi and other bacteria. The beta-mannanase gene could be expressed in Escherichia coli and the recombinant beta-mannanase was characterized by Western blot. This study provided a new source of carbohydrate hydrolysis enzyme with novel characteristics from Bacillus subtilis.

[Advance in biomechanical study of embryonic vascular system development].

Embryonic vascular system development is a complex process, whose progress is regulated by a variety of the stimulation and inhibition signals, and these signals must play synergistic effect so as to ensure that each stage of vascular development can proceed normally. The vascular development is controlled by the gene to a certain extent, and has received extensive attention. Recent studies have revealed the biomechanical role is necessary to embryonic vascular development, in which different mechanism of cell biomechanics is involved. In this review, we summarize the latest research progress on the role of biomechanical factors during embryonic vascular system development.

Protein tyrosine nitration in atherosclerotic endothelial dysfunction.

Accumulation of reactive oxygen species (ROS) can induce both protein tyrosine nitration and endothelial dysfunction in atherosclerosis. Endothelial dysfunction refers to impaired endothelium-dependent vasorelaxation that can be triggered by an imbalance in nitric oxide (NO) production and consumption. ROS reacts with NO to generate peroxynitrite, decreasing NO bioavailability. Peroxynitrite also promotes protein tyrosine nitration in vivo that can affect protein structure and function and further damage endothelial function. In this review, we discuss the process of protein tyrosine nitration, increased expression of nitrated proteins in cardiovascular disease and their association with endothelial dysfunction, and the interference of tyrosine nitration with antioxidants and the protective role in endothelial dysfunction. These may lead us to the conception that protein tyrosine nitration may be one of the causes of endothelial dysfunction, and help us gain information about the mechanism of endothelial dysfunction underlying atherosclerosis.

[Effects of different factors on transformed hairy root in Gynostemm apentaphyllum].

OBJECTIVE: To induce hairy roots of Gynostemm apentaphyllum by Agrobacterium rhizogenes strains. METHODS: Hairy roots were induced by the co-culture method of explants and Agrobacterium rhizogenes strains. Effects of different Agrobacterium rhizogenes strains, explants, pre(co)-culture time, Bacterial concentration, infecting time, As concentration and antibiotic medium on the transformation frequency were studied. RESULTS: The highest induction frequency was obtained form leaf 2 days co-cultivation, which were induced by Agrobacterium rhizogenes OD 600 0. 8 for 10 min, 100 micromol/L As and MS + 300 mg/L Cab. CONCLUSION: Hairy roots were induced by co-cultivation and the optimum induced condition were determined.

No Statistically Apparent Difference in Antiviral Effectiveness Observed Among Ribavirin Plus Interferon-Alpha, Lopinavir/Ritonavir Plus Interferon-Alpha, and Ribavirin Plus Lopinavir/Ritonavir Plus Interferon-Alpha in Patients With Mild to Moderate Coronavirus Disease 2019: Results of a Randomized, Open-Labeled Prospective Study.

BACKGROUND: Currently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread globally, causing an unprecedented pandemic. However, there is no specific antiviral therapy for coronavirus disease 2019 (COVID-19). We conducted a clinical trial to compare the effectiveness of three antiviral treatment regimens in patients with mild to moderate COVID-19. METHODS: This was a single-center, randomized, open-labeled, prospective clinical trial. Eligible patients with mild to moderate COVID-19 were randomized into three groups: ribavirin (RBV) plus interferon-α (IFN-α), lopinavir/ritonavir (LPV/r) plus IFN-α, and RBV plus LPV/r plus IFN-α at a 1:1:1 ratio. Each patient was invited to participate in a 28-d follow-up after initiation of an antiviral regimen. The outcomes include the difference in median interval to SARS-CoV-2 nucleic acid negativity, the proportion of patients with SARS-CoV-2 nucleic acid negativity at day 14, the mortality at day 28, the proportion of patients re-classified as severe cases, and adverse events during the study period. RESULTS: In total, we enrolled 101 patients in this study. Baseline clinical and laboratory characteristics of patients were comparable among the three groups. In the analysis of intention-to-treat data, the median interval from baseline to SARS-CoV-2 nucleic acid negativity was 12 d in the LPV/r+IFN-α-treated group, as compared with 13 and 15 d in the RBV+IFN-α-treated group and in the RBV+LPV/r+ IFN-α-treated group, respectively (p=0.23). The proportion of patients with SARS-CoV-2 nucleic acid negativity in the LPV/r+IFN-α-treated group (61.1%) was higher than the RBV+ IFN-α-treated group (51.5%) and the RBV+LPV/r+IFN-α-treated group (46.9%) at day 14; however, the difference between these groups was calculated to be statistically insignificant. The RBV+LPV/r+IFN-α-treated group developed a significantly higher incidence of gastrointestinal adverse events than the LPV/r+ IFN-α-treated group and the RBV+ IFN-α-treated group. CONCLUSIONS: Our results indicate that there are no significant differences among the three regimens in terms of antiviral effectiveness in patients with mild to moderate COVID-19. Furthermore, the combination of RBV and LPV/r is associated with a significant increase in gastrointestinal adverse events, suggesting that RBV and LPV/r should not be co-administered to COVID-19 patients simultaneously. CLINICAL TRIAL REGISTRATION: www.ClinicalTrials.gov, ID: ChiCTR2000029387. Registered on January 28, 2019.

Endogenous pH-responsive nanoparticles with programmable size changes for targeted tumor therapy and imaging applications.

Nanotechnology-based antitumor drug delivery systems, known as nanocarriers, have demonstrated their efficacy in recent years. Typically, the size of the nanocarriers is around 100 nm. It is imperative to achieve an optimum size of these nanocarriers which must be designed uniquely for each type of delivery process. For pH-responsive nanocarriers with programmable size, changes in pH (~6.5 for tumor tissue, ~5.5 for endosomes, and ~5.0 for lysosomes) may serve as an endogenous stimulus improving the safety and therapeutic efficacy of antitumor drugs. This review focuses on current advanced pH-responsive nanocarriers with programmable size changes for anticancer drug delivery. In particular, pH-responsive mechanisms for nanocarrier retention at tumor sites, size reduction for penetrating into tumor parenchyma, escaping from endo/lysosomes, and swelling or disassembly for drug release will be highlighted. Additional trends and challenges of employing these nanocarriers in future clinical applications are also addressed.

[Concentration polarization of low density lipoprotein at the distal end of carotid stenosis promotes atherogenesis].

To test the hypothesis that concentration polarization of atherogenic lipids may occur in the arterial system and play an important role in localization of atherosclerosis, we simulated and measured in vitro the luminal surface concentration of low density lipoprotein (LDL) in local stenosis at the distal end of carotid artery by number simulation and laser scanning confocal microscopy, then we designed carotid stenosis model to test the role of LDL concentration polarization in atherogenesis. The in vitro experiment showed that the luminal surface LDL concentration was higher than the bulk concentration as predicted by the concentration polarization theory. The relative luminal surface LDL concentration changed with the flow velocity and ratio of stenosis. The wall concentration of LDL was highest in the round tube with 40% stenosis at the same velocity, while the wall concentration of LDL was higher when Re was 250 than Re was 500 at the same extent of narrowness. The animal experiment also revealed that general atherogenic plaques obviously occurred at the distal end of local stenosis where concentration polarized. The results strongly support our hypothesis that concentration polarization of lipoproteins occurs in local stenosis at the distal end of carotid artery, and this in turn promotes the localization of atherosclerosis which develops in the arterial system.

[Analysis of signal peptides of the secreted proteins in Ralstonia solanacearum GMI1000].

The 3,440 open reading frames (ORFs) of Ralstonia solanacearum GMI1000 were used for predicting signal peptides by comprehensive analyses with SignalP 3.0, TMHMM 2.0, TargetP 1.01, LipoP 1.0 and PSORTb, and screening based on L value. Total 186 signal peptides with conserved amino acid residues are found and among them, 134 are secretary types, 22 are RR-motif types and 30 are lipoprotein signal peptides. The length distribution of signal peptide and its domains were analyzed systemically. No type pilin-like signal peptides and bacteriocins and pheronones signal peptides are found in the database of deduced proteins encoded by the genome of R. solanacearum GMI1000.

[The protection of elemene on injured HUVECs induced by hydrogen peroxide].

OBJECTIVE: To evaluate the effect of elemene on the survival of normal human umbilical endothelial cells (HUVECs) and the protection of elemene on injured HUVECs induced by hydrogen peroxide (H2O2). METHODS: Normal HUVECs were treated with elemene 1-100 microg/ml for 24-72 hours, the survival rate of HUVECs was determined by tetrazolium assay (MTT). To evaluate the protective effect of elemene on HUVECs from H2O2 injury, HUVECs were injured by 1 mmol/L H2O2 and then different final concentrations of elemene were added before the injury. After culturing 1 hour, then detecting the index of MDA, T-AOC, SOD, CAT, GSH-Px and NO. RESULTS: Elemene could inhabit the proliferation of VEC and it presented dose-dependent, while on the side of anti-oxidization injury, it also presented dose-dependent. MDA content and the effect of H2O2 to antioxidase activity were decreased, NO content in cell was increased, and the amount of apoptosis was reduced. CONCLUSION: Elemene has dual effects on the survival rate of normal HUVECs in vitro, which is related to the concentration and the duration of drug exposure. Elemene has protective effects on the injured endothelial cells injured by oxidization through the function of anti-lipid oxidization.

Hyperlipidemia-induced apoptosis of hippocampal neurons in apoE(-/-) mice may be associated with increased PCSK9 expression.

Hyperlipidemia is a risk factor for Alzheimer's disease (AD) and other neurodegenerative diseases. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a lipid regulatory gene involved in cell apoptosis. However, the function and mechanism of PCSK9 in neuronal apoptosis following hyperlipidemia remains to be elucidated. The present study established a hyperlipidemic mouse model by feeding a high­fat diet (HFD) to 6­week­old apoE(­/­) mice. Plasma lipid levels, hippocampal lipid accumulation, hippocampal histology, and hippocampal neuronal apoptosis were all monitored for changes. The expression levels of PCSK9, ß­secretase 1 (BACE1), B­cell lymphoma 2 (Bcl­2), Bcl­2­associated X protein (Bax), and caspase­3 in hippocampal CA3 and CA1 neurons were also measured. Results demonstrated that a HFD increased the lipid accumulation in the CA3 hippocampus and the levels of plasma lipids, including triglycerides, total cholesterol, low­density lipoprotein, and high­density lipoprotein. In addition, CA3 neurons in the HFD group indicated apparent injuries and increased neuronal apoptosis, which are associated with the expression of Bcl­2, Bax, and caspase­3. A HFD also increased the expression levels of PCSK9 and BACE1. BACE1 promotes cleavage of amyloid precursor proteins to generate ß­amyloid peptide (Aß), which induces neuronal apoptosis. Protein levels of Aß are associated with the observation of amyloid plaques in the hippocampus of the HFD group. The results suggest that hyperlipidemia regulates neuronal apoptosis by increasing PCSK9 and BACE1 expression. Overall, the current study may elucidate the role of lipid metabolism disorder in AD pathogenesis.

Elevating VEGF-A and PDGF-BB secretion by salidroside enhances neoangiogenesis in diabetic hind-limb ischemia.

Hind-limb ischemia (HLI) is one of the major complication of diabetic patients. Angiogenesis potential in diabetic patients is severely disrupted, and the mechanism underlying it has not been fully elucidated, making it an obstacle for developing an efficient therapeutic angiogenesis strategy. Skeletal muscle cells, through their paracrine function, had been known to be critical for neoangiogenesis. Here we found that hyperglycemia upregulates the expression of skeletal muscle cells prolyl hydroxylase domain 3 (PHD3), which resulted in the decrease of the secretion of angiogenic factors, especially VEGF-A and PDGF-BB. We showed that treatment with salidroside, a small molecule drug, significantly suppresses PHD3 expression and increases VEGF-A and PDGF-BB secretion from skeletal muscle cells, which in turn enhances the proliferation and migration potentials of endothelial and smooth muscle cells. Finally, we demonstrated that intramuscular injection of salidroside into the ischemic hind limbs of diabetic HLI model mice could efficiently induce neoangiogenesis and blood perfusion recovery. Thus, our novel findings not only reveal the effects of hyperglycemia on the angiogenesis potential of skeletal muscle cells and the mechanism underlying it, but also provides a novel finding suggesting that salidroside might be a potential small molecule drug for diabetic HLI.

Dual tumor-targeted poly(lactic-co-glycolic acid)-polyethylene glycol-folic acid nanoparticles: a novel biodegradable nanocarrier for secure and efficient antitumor drug delivery.

Further specific target-ability development of biodegradable nanocarriers is extremely important to promote their security and efficiency in antitumor drug-delivery applications. In this study, a facilely prepared poly(lactic-co-glycolic acid) (PLGA)-polyethylene glycol (PEG)-folic acid (FA) copolymer was able to self-assemble into nanoparticles with favorable hydrodynamic diameters of around 100 nm and negative surface charge in aqueous solution, which was expected to enhance intracellular antitumor drug delivery by advanced dual tumor-target effects, ie, enhanced permeability and retention induced the passive target, and FA mediated the positive target. Fluorescence-activated cell-sorting and confocal laser-scanning microscopy results confirmed that doxorubicin (model drug) loaded into PLGA-PEG-FA nanoparticles was able to be delivered efficiently into tumor cells and accumulated at nuclei. In addition, all hemolysis, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, and zebrafish-development experiments demonstrated that PLGA-PEG-FA nanoparticles were biocompatible and secure for biomedical applications, even at high polymer concentration (0.1 mg/mL), both in vitro and in vivo. Therefore, PLGA-PEG-FA nanoparticles provide a feasible controlled-release platform for secure and efficient antitumor drug delivery.

[Development of drug coating on drug eluting stents].

Researches on drug-eluting stents are now focusing on three main aspects: the stent materials, the coating matrix material and the selection, adhesion and controlled release of the biological agents. The current development progresses of the coating materials, their characteristics, and the coating method for metallic stents are reviewed in this paper.

[Study on surface modification and biocompatibility of NiTi alloy intravascular stents].

This paper introduces the surface modification of NiTi alloy intravascular stents for roughness by chemical erosion and plasma deposition technology. The stent which had been granulated with chemical erosion was treated with TiO2 film prepared with Gel-sol. The study on the biocompatibility of the modified stent by the above two ways shows that the modified stent is rougher, and its anticoagulation and hydrophilicity are improved. However, the capability of erosion resistance is not enhanced significantly.

[In vitro study of platelet glycoprotein monoclonal antibody eluting stents].

In order to prove the feasibility of preparation of the drug-incorporated stent by immersing stent wires in the monoclonal antibody (mAb) solution, fluorescence stain and image analysis were used to evaluate the L-PLA-coated stent. Absorption was measured using a radioisotope technique after preparing the mAb-incorporated stent, and the absorption curve was determined from the absorption data. In an in vitro perfusion circuit, the antibody was eluted from the stent matrices, and the related influence factors were evaluated based on the release data.