Eu MOF-enhanced FeNCD nanozymes for fluorescence and highly sensitive colorimetric detection of tetracycline.

The constrained enzymatic activity and aggregation challenges encountered by small-sized nanozymes pose obstacles to their practical utility, necessitating a strategy to mitigate aggregation and boost enzymatic catalytic efficiency. In this work, a negatively charged Eu MOF was utilized as the encapsulation matrix, encapsulating the small-sized nanozymes FeNCDs into the Eu MOF to synthesize an FeNCDs@Eu MOF. The dispersibility of the encapsulated FeNCDs was increased, and owing to the negative charge of the FeNCDs@Eu MOF, electrostatic pre-concentration of the positively charged target molecule tetracycline (TC) was facilitated, thereby amplifying the enzymatic catalytic efficiency of the FeNCDs. The response of the FeNCDs to TC increased by nearly 6 times upon encapsulation. The TC detection limit (LOD) of the FeNCDs@Eu MOF-based sensor is as low as 11.63 nM. The incorporation of fluorescence detection expanded the linear range of the sensor, rendering it more suitable for practical sample detection.

Analysis of influencing factors on monohydroxylated derivative of oxcarbazepine plasma concentration in children with epilepsy.

PURPOSE: This study aimed to investigate the factors affecting the plasma concentration of monohydroxylated derivative (MHD) of oxcarbazepine (OXC) in children with epilepsy. METHODS: We recruited 125 children with epilepsy who received OXC monotherapy. Among them, 16 single nucleotide polymorphisms were detected by MassARRAY genotyping technology to evaluate the influence of related factors on the plasma concentration of OXC monotherapy. MHD is the main active metabolite of OXC, and its plasma concentration was measured by high-performance liquid chromatography (HPLC). RESULTS: Bivariate correlation analysis revealed that concentration-dose ratio (CDR) increased with weight, and the corresponding maintenance dose decreased with weight (r=0.317, P=0.001 for CDR; r=-0.285, P=0.000 for OXC maintenance dose). The duration of seizure was found to be associated with CDR (0.90 ± 0.36 vs 0.74 ± 0.26 µg·kg/mg/mL for ≥6 years vs <1 year, P=0.028; 0.90 ± 0.36 vs 0.64 ± 0.21 µg·kg/mg/mL for ≥6 years vs 1-3 years, P=0.004; 0.90 ± 0.36 vs 0.69 ± 0.18 µg·kg/mg/mL for ≥6 years vs 3-6 years, P=0.031). The CDR of patients with ABCB1 rs1045642 mutation homozygous GG type is higher than heterozygous AG type (0.79 ± 0.30 vs 0.68 ± 0.20 µg·kg/mg/mL for AG vs GG, P=0.032). CONCLUSION: This study clarified the association of weight, duration of seizure, and gene polymorphisms of ABCB1 rs1045642 with MHD plasma concentration in children with epilepsy.

The Impact of hyperuricemia on long-term clinical outcomes of renal transplant recipients: a systematic review and meta-analysis.

PURPOSE: To evaluate the effect of hyperuricemia on clinical outcomes of renal transplant recipients (RTRs). METHODS: A literature search of PubMed, Cochrane, Embase was conducted up to March 20, 2020. The primary outcome was the estimated glomerular filtration rate (eGFR). The second outcomes were the risk of graft loss, death, cardiovascular event and the level of triglyceride. The following search terms were utilized: ((Hyperuricemic group) OR (Hyperuricaemia) OR (Hyperuric) OR (Urea acid) OR (Uric acid) OR (Acid urate) OR (Urate) OR (Gout)) and ((Transplantation) OR (Transplantations) OR (Transplant) OR (Transplants) OR (Graft)). RESULTS: 28 studies with 18224 patients were eligible for inclusion. There was no significant difference in eGFR (<12 months, p=0.07), the risk of graft loss (<60 months, p=0.07) and death (<60months, p=0.19) between the hyperuricemic and normouricemic group in the early post-transplantation period. But increased uric acid levels contributed to the long-term decline of eGFR, the risk of graft loss and death increased after transplantation. Hyperuricemia increased the risk of cardiovascular event with no significant difference in the level of triglyceride between the two groups. CONCLUSIONS: Increased uric acid levels contributed to the long-term decline of eGFR, increased risk of graft loss and death after transplantation. Although there was no significant effect on triglyceride, hyperuricemia increased the risk of cardiovascular event.

Corrigendum: Biosynthesis, characterization, and antifungal activity of plant-mediated silver nanoparticles using Cnidium monnieri fruit extract.

[This corrects the article DOI: 10.3389/fmicb.2023.1291030.].

Highly sensitive biosensor for specific miRNA detection based on cascade signal amplification and magnetic electrochemiluminescence nanoparticles.

Herein, magnetic electrochemiluminescence (ECL) nanoparticle Fe3O4@PtPd/Ru(bpy)32+ had been synthesized then been coupled with CRISPR/Cas13a system and Zn2+ dependent DNAzyme to design a novel ECL biosensor for specific detection of microRNA-145 (miRNA). The synthesized multifunctional magnetic nanoluminescent materials Fe3O4@PtPd/Ru(bpy)32+ not only load Ru(bpy)32+ to provide ECL signals, but also can quickly achieve separation and enrichment from complex matrices. In addition, ferrocene (Fc) was used as a quencher in the Ru(bpy)32+/tripropylamine (TPA) system. Fc was modified on DNA bound to Fe3O4@PtPd. Benefited from the highly specific recognition ability of CRISPR/Cas13a, the target miRNA induces CRISPR/Cas13a trans-cleavage to trigger the Zn2+-dependent DNAzyme cyclic cleavage to realize the dual signal amplification. DNA modified by Fc was split by target miRNA-induced cleaving, and then magnetic separation was performed to keep Fc away from the surface of the nanoparticles. Thus, the enhanced ECL signal was obtained to detect miRNA-145. Under optimized conditions, the prepared sensor showed a wide linear range (1 fM to 1 nM) and a low limit of detection (LOD) down to 0.41 fM. Furthermore, it shows excellent selectivity and good reproducibility. The proposed ECL platform has huge potential applications in the development of various sensitive sensors for detecting the other miRNA.

Etiology and outcomes of acute kidney injury in Chinese children: a prospective multicentre investigation.

BACKGROUND: The incidence of AKI appears to have increasing trend. Up to now, prospective, multi-center, large-sample epidemiological study done on pediatric AKI on aspects of epidemiological characteristics, causes and outcomes have not reported. It is necessary to develop prospective, multi-center, large-sample epidemiological study in our country on pediatric AKI. The aim of this study was to determine the clinical features, etiology, and outcomes of acute kidney injury (AKI) in Chinese children. METHOD: Paediatric patients (≤18 years old) admitted to 27 hospitals (14 children's hospitals and 13 general hospitals) affiliated with the Medical University were investigated. AKI was defined using the 2005 Acute Kidney Injury Network criteria. RESULTS: During the study period, 388,736 paediatric patients were admitted. From this total, AKI was diagnosed in 1,257 patients, 43 of whom died. The incidence and mortality of AKI was 0.32% and 3.4% respectively. The mean (± SD) age of patients was 48.4 ± 50.4 months. Among the 1,257 AKI paediatric patients, 632 were less than one year old. Among the AKI paediatric patients, 615 (48.9%) were in stage 1, 277 (22.0%) in stage 2, and 365 (29.0%) in stage 3. The most common causes of AKI were renal causes (57.52%), whereas postrenal (25.69%) and prerenal (14.96%) causes were the least common. The three most common causes of AKI according to individual etiological disease were urolithiasis (22.35%), of which exposure to melamine-contaminated milk accounted for the highest incidence (63.7%); acute glomerulonephritis (10.10%); and severe dehydration (7.48%). A total of 43 AKI patients (3.4%) died during their hospital stay; 15 (34.9%) of the 43 died as a result of sepsis. CONCLUSION: Primary renal diseases are a major risk factor for paediatric AKI in China. In terms of specific etiological disease, urolithiasis (postrenal disease) was the leading cause of paediatric AKI in 2008, when the disease was linked to exposure to melamine-contaminated milk. Sepsis is the leading cause of death in Chinese paediatric AKI patients. Future studies should focus on effective ways of controlling renal disorders and sepsis to improve the clinical management of paediatric AKI in China.

Biosynthesis, characterization, and antifungal activity of plant-mediated silver nanoparticles using Cnidium monnieri fruit extract.

The present study describes a novel method for green synthesis of silver nanoparticles using Cnidium monnieri (CM-AgNPs). Cnidium monnieri fruit is an excellent anti tinea drug that can be used externally to treat superficial fungal infections in the human body. The aqueous ethanolic extract of Cnidium monnieri fruit was prepared and employed in the synthesis of stable silver nanoparticles via biological reduction method. The synthesis conditions of CM-AgNPs was systematically optimized using Box-Behnken design. CM-AgNPs were well characterized by UV-spectroscopy and X-ray powder diffraction (XRD), and it was confirmed that the synthesized particles were AgNPs. The possible functional groups required for the reduction and stabilization of CM-AgNPs in the extract were identified through FTIR spectrum. The size of CM-AgNPs structure was confirmed to be approximately 44.6 nm in polydisperse spherical shape through scanning electron microscopy (SEM), transmission electron microscopy (TEM), and laser dynamic light scattering (DLS). Further, the minimum inhibitory concentration 90% (MIC90) ratios values of Cm-AgNPs against Trichophyton rubrum (7 d), T. mentagrophytes (7 d) and Candida albicans (24 h) were 3.125, 3.125, and 0.78125 µg/mL, respectively, determined by the broth micro dilution method. Finally, the result was concluded that the synthesized AgNPs could be further evaluated in large scale as a potential human topical antifungal agent.

Controllable release ratiometric fluorescent sensor for hyaluronidase via the combination of Cu2+-Fe-N-C nanozymes and degradable intelligent hydrogel.

As a popular controllable-released carrier, intelligent hydrogels are often used in drug delivery and disease therapeutics. Meanwhile, benefit from the mimic-enzyme activity performance, Fe-N-C nanozymes have been widely used in sensing and analysis. However, the combination of intelligent hydrogels with specific degradability and Fe-N-C nanozymes with enhanced activity in one system to achieve controllable and sensitive detection is rare. Herein, we combine intelligent hydrogel with mimic peroxidase activity enhanced Fe-N-C nanozymes to construct a ratiometric fluorescence probe for sensitive detection of hyaluronidase (HAase). The modification of copper ions has been proved to enhance the mimic enzyme activity of Fe-N-C nanozymes greatly. Cu2+ modified Fe-N-C nanozymes were embedded in hyaluronic acid hydrogel. In the presence of HAase, the HA hydrogel structure was hydrolyzed and released Cu2+-Fe-N-C nanozymes gradually. The released Cu2+-Fe-N-C nanozymes are used to catalyze the hydrogen peroxide system so that o-phenylenediamine is oxidized to orange fluorescent 2, 3-diaminophenolazine (DAP). Due to the electrostatic interaction, the fluorescence resonance energy transfer can occur between the negatively charged copper nanoclusters emitted by 430 nm and the positively charged DAP emitted by 560 nm. The activity of HAase was monitored according to the ratio of fluorescence intensity at 560 nm and 430 nm (F560/F430). The linear range of this method is 0-10.0 U/ml and the detection limit is 0.43 U/mL (S/N = 3). This strategy has been further applied to biological samples successfully.

Effect of octreotide-polyethylene glycol(100) monostearate modification on the pharmacokinetics and cellular uptake of nanostructured lipid carrier loaded with hydroxycamptothecine.

A new conjugate, octreotide-polyethylene glycol(100) monostearate (OPMS), was developed for the enhancement of targeting delivery of hydroxycamptothecine (HCPT) loaded in nanostructured lipid carrier (NLC). 2 × 10(-3) and 5 × 10(-3) mmol of OPMS were respectively used to modify NLC so that the targeted nanocarriers with low and high ligand density were obtained. For comparison, the pegylated NLCs without octreotide were prepared by adding equal molar amounts of polyethylene glycol(100) monostearate (PGMS). The relation between the modification levels and properties of various NLCs were studied in vivo and in vitro. At a high modification level, a slower release rate of HCPT and the more stable nanocarriers was achieved. At the same time, the fixed aqueous layer thickness (FALT) and average surface density of PEG chains (SD(PEG)) was increased, but the distance (D) between two neighboring PEG grafting sites became narrower. The in vivo pharmacokinetic study in healthy rat indicated that the modified NLCs had a longer circulation than NLC (P < 0.05) due to pegylation effect and OPMS modified NLCs had larger MRT and AUC(0-t) than that of PGMS modified NLCs at the same modification level. Furthermore, the florescence microscopy observation also showed the targeting effect of octreotide modification on somatostatin receptors (SSTRs) of tumor cell (SMMC-7721). The uptake of SMMC-7721 was much more than that of normal liver cell (L02) for OPMS modified NLC, and the highest uptake was observed for 5 × 10(-3) mmol of OPMS modified one. No obvious difference was found among the L02 uptake of OPMS modified NLCs and NLC, but their uptake was higher than that of PGMS modified NLCs. All the results indicated that the OPMS highly modified NLCs would improve the effect of antitumor therapy by inhibiting the degradation, evading RES and enhancing the drug uptake of tumor cells.

Differences in tissue expression of HBV markers in children with HBV-associated glomerulonephritis.

BACKGROUND/AIMS: Hepatitis B virus-associated glomerulonephritis (HBV-GN) is recognized as one of the major secondary nephropathies in HBV high-risk areas. To determine possible differences in the expression of HBV immune markers in tissues, we retrospectively examined HBV immune markers in the serum, renal tissues, and liver tissues in 132 HBV-GN children. METHODS: All 132 patients had biopsy-proven HBV-GN including the presence of positive HBV antigens in the kidney. Serum-HBV immune markers were tested by an enzyme-linked immunosorbent assay. Renal and liver biopsies were done in 26 patients. All renal tissues were examined for HBV immune markers by immunofluorescence, and liver tissues were examined by immunohistochemistry. RESULTS: Among the 132 patients, all showed varying degrees of kidney injury. Serum hepatitis B envelope antigen (HBeAg) was positive in 80 patients and negative in 52 patients. The positivity rate of Hepatitis B core antigen in renal tissue was statistically higher in serum HBeAg (-) than in serum HBeAg (+) patients (96.2% vs. 55.0%). Furthermore, there was no relationship between the presence of hepatitis B surface antigen and HBcAg in liver and renal tissue. CONCLUSION: HBV markers are not consistently present in serum, renal tissues, and liver tissues in children with HBV-GN.

Box-Behnken optimization design and enhanced oral bioavailability of thymopentin-loaded poly (butyl cyanoacrylate) nanoparticles.

This study was done to prepare thymopentin (TP5)-loaded poly (butyl cyanoacrylate) nanoparticles (TP5-PBCA-NPs) and evaluate thier efficacy for oral delivery. TP5-PBCA-NPs were prepared by emulsion polymerization, and the formulation was optimized based on Box-Behnken experimental design. The physico-chemical characteristics of TP5-PBCA-NPs were evaluated using transmission electron microscopy (TEM), malvern zetasizer, Fourier transform infra-red spectroscopy (FT-IR) and differential scanning calorimetry (DSC). The encapsulation efficiency, enzymatic degradation and release behavior of TP5-PBCA-NPs in various media were evaluated using a high-performance liquid chromatography (HPLC) method. The pharmacodynamic studies on oral administration of TP5-PBCA-NPs were performed in FACScan flow cytometry. An optimum formulation consisted of 0.7% poloxamer 188 (Pol), 0.6% dextran-70 (Dex), 0.1% sodium metabisulfite (Sm), 0.1% TP5 and 1% (v/v) n-butyl cyanoacrylate. The particle size and zeta potential of optimized TP5-PBCA-NPs was 212 nm and -22.6 mV respectively with 82.45% encapsulation efficiency. TP5 was entrapped inside the nanoparticles in molecular dispersion form. The release of TP5 from PBCA-NPs was pH dependent; the cumulative release percentage in 0.1 M HCI for 4 hours was less than 16% while it was more than 80% in pH6.8 PBS. The PBCA-NPs could efficiently protect TP5 from enzymatic degradation; the remained percentage of TP5 encapsulated in PBCA-NPs was 58.40% after incubated with trypsin in pH6.8 PBS for 4 h while it was only 32.29% for free drug. In the oral administration study in vivo, the lowered T-lymphocyte subsets values were significantly increased and the raised CD4+/CD8+ ratio was evidently reduced compared with that of TP5 solution (p < 0.05), and the improvement of bioavailability was dose-dependent. These results indicated that the PBCA nanoparticles may be a promising carrier for oral delivery of TP5.

High-Resolution Microscopy to Learn the Nuclear Organization of the Living Yeast Cells.

The spatial organization of the nucleus is a key determinant in all genome activities. However, the accurate measurement of the nuclear organization is still technically challenging. Here, the technology NucQuant we created previously was utilized to detect the variation of the nuclear organization, including the heterogeneity of the nuclear geometry, the change of the NPC distribution along different cell cycle stages during interphase, and the organization of the nucleolus. The results confirmed that not only the growth rate and the NPC distribution are influenced by the carbon source; the nuclear shape is also impacted by the carbon source. The nuclei lost their spherical geometry gradually when the cell was cultured from the most to a less favorable carbon source. We also discovered that the nucleolus prefers to locate at the nuclear periphery, which was called the "genes poor region," especially when the cells entered quiescence. Furthermore, the distribution of the NPC along the different stages during the interphase was analyzed. We proposed that with the growth of the cell, the nucleus would grow from the surface of the NE flanking the nucleolus firstly.

[Preliminary analysis of urinary proteomics in children with steroid-resistant and steroid-sensitive nephrotic syndrome].

OBJECTIVE: To study and identify the protein markers in the urine of children with steroid-sensitive (SSNS) and steroid-resistant nephrotic syndrome(SRNS). METHODS: Total urinary proteins were extracted from children with SSNS before and after steroid therapy, SRNS, and healthy children (n=5 in each group). Urinary proteins were separated by immobilized pH gradient based on two-dimensional gel electrophoresis (2-DE). The silver-stained 2-DE gels were scanned with digital Image Scanner and analyzed with Image Master 2-DE Elite 3.01 software. Peptide mass fingerprint (PMF) of differential protein spots was obtained with MALDI-TOF-MS. Proteins were identified by Mascot software based on NCBI protein database. RESULTS: There were 66 spots with different expression of protein between SRNS children and SSNS children before steroid therapy, and 24 spots and 27 spots only occurred in SRNS children and SSNS children before steroid therapy, respectively. There were 75 spots with different expression of protein between SSNS children after steroid therapy and healthy controls, and 11 spots only occurred in SSNS children after steroid therapy. Eighteen protein spots with different expression (6 spots in each nephrotic group) were chose and analyzed by MALDI-TOF-MS, and 9 types of proteins were identified. CONCLUSIONS: Nine types of urinary proteins with different expression (6 spots in each nephrotic group) were identified between SRNS and SSNS children, and they might be the biomarkers for SRNS or SSNS.

Ubiquitin specific peptidase 5 regulates colorectal cancer cell growth by stabilizing Tu translation elongation factor.

Ubiquitin specific peptidase 5 (USP5) is a ubiquitous expressed deubiquitinating enzyme (DUB). It has been shown involved in DNA repair, apoptosis, inflammation, and tumor cell growth. However, the function and molecular mechanism of USP5 in colorectal cancer (CRC) are still unclear. In the present study, we asked how it affected the growth of colorectal cancer cells. Methods: A shRNA-based high-content screening was performed to identify DUBs affecting the growth of CRC cells. CCK-8 assay and xenografts were used to assess CRC cell growth, survival and tumorigenesis. RT-qPCR, immunoblotting and immunohistochemistry were carried out to quantitate USP5 expression in CRC tissues and cell lines. Immunoprecipitation and mass spectrometry analysis were performed to identify USP5-interacting proteins. Cycloheximide chase was performed to assess Tu translation elongation factor (TUFM) stability. Dual luciferase reporter assay was utilized for USP5 promoter analysis. Results: We found that USP5 was highly expressed in a group of primary CRC tissues, and the increased USP5 was correlated with clinical stages and shorter overall survival. While USP5 knockdown effectively inhibited CRC cell growth, overexpressed USP5 promoted the growth of CRC cells and made them more resistant to doxorubicin (DOX). TUFM was discovered as a substrate of USP5. USP5 deubiquitinated TUFM and increased its level in CRC cells. Enforced expression of TUFM was able to alleviate the growth inhibition induced by USP5 knockdown. Further analyses showed that EBF transcription factor 1 (EBF1) was a major regulator for USP5 transcription, and DOX inhibited EBF1-USP5-TUFM axis in CRC cells. Conclusions: USP5 was required for CRC cells and promoted their growth and resistance to chemotherapeutics. TUFM was a USP5 deubiquitinating substrate that mediated the cellular effects of USP5. The transcription of USP5 was regulated by EBF1. Thus, targeting EBF1-USP5-TUFM axis is a potential novel strategy for CRC treatment.

Preparation and Characterization of Silymarin Synchronized and Sustained Release Dropping Pill.

PURPOSE: This study aimed to develop a synchronized and sustained-release silymarin dropping pill, and to evaluate its pharmacokinetic characteristics. METHOD: Polyoxyethylene stearate, glyceryl monostearate, and stearic acid were used to prepare the dropping pills. X-ray powder diffraction, differential scanning calorimetry, and release were used to evaluate its physicochemical properties. The plasma concentration of silybin in beagle dogs after oral administration of silymarin dropping pills and silymarin capsule was determined by RP-HPLC. RESULTS: Synchronized release was achieved with high similarity factor f2 values between every set of two of the five components. Mean plasma concentration-time curves of silymarin after oral administration of dropping pills in beagle dogs were in accordance with first-order absorption and open twocompartment model. The Tmax, Cmax, and AUC0-∞ of dropping pills in beagle dogs were 0.8750±0.13 h, 0.8183±0.07 µg·ml-1, and 2.274±0.90 µg·h·ml-1, respectively. Silymarin dropping pills prolonged in vivo exposure and reduced maximum in vivo concentration, achieving a stable level in the serum. CONCLUSION: The combination of solid dispersion technique and dropping pill formulation allowed synchronized release of multiple components in herbal medicine, and has potential application in the development of sustained release in herbal medicine.

Preparation and characterization of silymarin synchronized-release microporous osmotic pump tablets.

The pharmacological activity of herbal medicine is an overall action of each component in accordance with their original proportion. An efficient, sustained, and controlled-release drug delivery system of herbal medicine should ensure the synchronized drug release of each active component during the entire release procedure. In this study, silymarin (SM), a poorly soluble herbal medicine, was selected as a model drug to develop a synchronized-release drug delivery system: an SM microporous osmotic pump (MPOP) tablet. The SM was conjugated with phospholipid (SM phytosome complex, SM-PC) to improve the solubility, and the difference in the apparent octanol-water partition coefficient between the two components was significantly reduced. The dissolution rate of SM-PC was significantly higher than SM active pharmaceutical ingredients and was the same as that of the commercial SM capsule. The SM-PC was used to generate the MPOP tablet. SM was mixed with poly(ethylene) oxide and sodium chloride (an osmotic agent) to form the MPOP core, followed by coating with cellulose acetate and poly(ethylene) oxide to generate the SM MPOP. The results demonstrated that SM MPOP could synchronically and sustainably release the five active components within 12 hours (the similar coefficient f 2 between two components was >65), and the average cumulative release rate was 85%. Fitting of the drug-release curve showed a zero-order release profile for SM MPOP. Our study showed that the phytosome complex technique combined with the MPOP system will achieve synchronized release of the various active components of herbal medicine and have potential applications in developing sustained release preparations in herbal medicine.

Development of solidified self-microemulsifying delivery systems with enhanced stability of sirolimus and extended release.

The application of sirolimus (SRL) as immunosuppressive agent is hampered by its poor water solubility and narrow therapeutic range. The self-microemulsifying drug delivery system (SMEDDS) succeeded in improving the solubility of SRL in our previous work. In this study, the formulation of the SMEDDS was further optimized by investigating the influence of the excipients including the media, antioxidant and organic acid. It was demonstrated that addition of 0.20% of citric acid in SMEDDS most efficiently promoted the stability of SRL under high temperature (40±2°C), high humidity (relative humidity 90±5%) or strong light irradiation (4500±500lx). SMEDDS absorbed by microcrystalline cellulose (MCC) was mixed with hydroxypropyl methylcellulose (HPMC) to prepare tablets. The optimal formulation composed of 15% of HPMC 100 LV with hardness of 120N, which had a sustained release of 12h. Results of X-ray powder diffraction and differential scanning calorimetry demonstrated that SRL in the tablets was in amorphous or molecularly dispersed state. The SMEDDS-tablets presented as promising substrates for water insoluble drugs with enhanced stability and extended release.

Oral absorption enhancement of salmon calcitonin by using both N-trimethyl chitosan chloride and oligoarginines-modified liposomes as the carriers.

In this study both N-trimethyl chitosan chloride (TMC) and oligoarginine (Arg8) were utilized to modify liposomes as the multifunctional carriers (TMC-Arg8-Lips) for enhancing the oral absorption of salmon calcitonin. Two permeation enhancers with positive charges were sequentially adsorbed on the liposomal surface with negative charges by electrostatic interaction. Instead of salmon calcitonin, fluorescein isothiocyanate dextran (FD4) was loaded in TMC-Arg8-Lips for Caco-2 cell permeation test in vitro and penetration examination in rat intestinal tract in vivo. The results showed that the apparent permeability coefficient (Papp) of TMC-Arg8-Lips containing FD4 were 7.0-, 4.4-, 1.8- and 1.4-folds higher than FD4 solution, FD4-TMC solution, non-modified liposomes (Non-Lips) and TMC modified liposomes (TMC-Lips), respectively. A strong fluorescence was observed by confocal laser scanning microscope (CLSM) at rat intestinal wall isolated in different times after the FD4 loaded carriers were intragastrically administrated. Furthermore, the images revealed that TMC-Arg8-Lips could penetrate deeply inside the mucosal membrane. The pharmacodynamic study indicated that TMC-Arg8-Lips containing calcitonin were more efficient in enhancing the absorption and prolonging the reduction of blood calcemia in rats. The area above the plasma calcium concentration-time curve (AAC) of TMC-Arg8-Lips containing calcitonin was increased by more than 16.6- and 1.6-fold when compared to Non-Lips and TMC-Lips, respectively.

The potential use of allogeneic platelet-rich plasma for large bone defect treatment: immunogenicity and defect healing efficacy.

Autologous platelet-rich plasma (PRP) has been extensively investigated for large bone defect treatment, but its clinical application is harassed by controversial outcome, due to highly variable PRP quality among patients. Alternatively, allogeneic PRP from well-characterized donors cannot only generate more consistent and reliable therapeutic effect but also avoid harvesting large quantities of blood, an additional health burdens to patients. However, the use of allogeneic PRP for bone defect treatment is generally less investigated, especially for its immunogenicity in such application. Here, we meticulously investigated the immunogenicity of allogeneic PRP and evaluated its healing efficacy for critical-sized defect treatment. Allogeneic PRP contained 4.1-fold and 2.7- to 4.9-fold higher amount of platelets and growth factors than whole blood, respectively. The intramuscular injection of allogeneic PRP to rabbits did not trigger severe and chronic immunoresponse, evidenced by little change in muscular tissue microstructure and CD4⁺/CD8⁺ T lymphocyte subpopulation in peripheral blood. The implantation of allogeneic PRP/deproteinized bone matrix (DPB) constructs (PRP+DPB) successfully bridged 1.5-cm segmental radial defects in rabbits, achieving similar healing capacity as autologous MSC/DPB constructs (MSC+DPB), with greater bone formation (1.1-1.5×, p<0.05) and vascularization (1.3-1.6×, p<0.05) than DPB alone, shown by histomorphometric analysis, bone mineral density measurement, and radionuclide bone imaging. Furthermore, the implantation of both allogeneic PRP- and autologous MSC-mediated DPB constructs (PRP + MSC + DPB) resulted in the most robust bone regeneration (1.2-2.1×, p<0.05) and vascularization (1.3-2.0×, p<0.05) than others (PRP+DPB, MSC+DPB, or DPB alone). This study has demonstrated the promising use of allogeneic PRP for bone defect treatment with negligible immunogenicity, great healing efficacy, potentially more consistent quality, and no additional health burden to patients; additionally, the synergetic enhancing effect found between allogeneic PRP and autologous MSCs may shed a light on developing new therapeutic strategies for large bone defect treatment.

Octreotide-mediated tumor cell uptake and intracellular pH-responsive drug delivery of the self-assembly supramolecular nanocarrier.

In this study, DOX-loaded supramolecular nanocarrier (DLSC) was assembled by using two new amphiphilic polymers, octreotide-polyethylene glycol monostearate (OPMS) and N-octyl-N-succinyl O-carboxymethyl chitosan (OSCC). The characteristics of the DLSC were investigated. The results indicated that the significant pH-triggered release in vitro. The cellular uptake of DLSC was much higher than that of DOX-loaded OSCC micelles (DLOM) in the SMMC-7721 (somatostatin receptor (SSTR) over-expressed cell) cells, which suggested the SSTR-mediated properties. A considerable amount of drug entered the nucleus due to the pH-triggered deformation of the supramolecular structure and rapid release of drug in acidic endosomes of tumor cells. The killing efficacy was much higher than that of DLOM in the SMMC-7721. In S180 sarcoma-bearing KM mice, the biodistribution and therapeutic activity were studied. DLSC showed extended circulation time in plasma, decreasing drug concentrations in the heart and accumulating drug concentrations in the pancreas and tumor. In addition, minimized weight changes and cardiac toxicity, high suppression ratio of tumor growth and longer survival time were observed after intravenous injection of DLSC. The studies suggested that the supramolecular nanocarrier constructed of different designated polymers with multiple functions would be one of the most effective approaches for active targeting drug delivery.