Can nanoparticles and nano‒protein interactions bring a bright future for insulin delivery?

Insulin therapy plays an essential role in the treatment of diabetes mellitus. However, frequent injections required to effectively control the glycemic levels lead to substantial inconvenience and low patient compliance. In order to improve insulin delivery, many efforts have been made, such as developing the nanoparticles (NPs)-based release systems and oral insulin. Although some improvements have been achieved, the ultimate results are still unsatisfying and none of insulin-loaded NPs systems have been approved for clinical use so far. Recently, nano‒protein interactions and protein corona formation have drawn much attention due to their negative influence on the in vivo fate of NPs systems. As the other side of a coin, such interactions can also be used for constructing advanced drug delivery systems. Herein, we aim to provide an insight into the advance and flaws of various NPs-based insulin delivery systems. Particularly, an interesting discussion on nano‒protein interactions and its potentials for developing novel insulin delivery systems is initiated.

Curcumin- and Cyclopamine-Loaded Liposomes to Enhance Therapeutic Efficacy Against Hepatic Fibrosis.

BACKGROUND AND PURPOSE: Hepatic fibrosis is a public health problem characterized by activation of hepatic stellate cells (HSCs), which triggers excessive production of extracellular matrix (ECM). Inhibition of HSC activation may be an effective treatment. Since various pathways control HSC activation, a combination of drugs with different mechanisms may be more effective than monotherapy. METHODS: Here, we prepared liposomes loaded with curcumin and cyclopamine to inhibit HSC activation. We systematically analyzed the physicochemical characteristics of liposomes loaded with the two drugs, as well as their effects on HSC proliferation, activation and collagen production on gene, protein and cellular levels. RESULTS: The prepared liposomes helped solubilize both drugs, contributing to their uptake by cells. Liposomes loaded with both drugs inhibited cell proliferation, migration and invasion, as well as induced more apoptosis and perturbed the cell cycle more than the free combination of both drugs in solution or liposomes loaded with either drug alone. Liposomes loaded with both drugs strongly suppressed HSC activation and collagen secretion. CONCLUSION: Our results suggest that liposome encapsulation can increase the uptake of curcumin and cyclopamine as well as the synergism between them in anti-fibrosis. This approach shows potential for treating hepatic fibrosis.

The UV absorption of graphene oxide is size-dependent: possible calibration pitfalls.

Graphene oxide (GO) is often quantified via its UV absorption, typically at around 230 nm. This is convenient but the effect of the size of GO on the accuracy of this method has been ignored so far. The authors report that the molar absorbance of GO is size-dependent. Data are presented on the absorbance of small (hydrodynamic diameter 1 µm), medium sized (1.5 µm), and large (2.2 µm) GO particles at wavelengths of 210, 230 and 250 nm. In general, linear relationship and good regression fits are obtained, but with different slope depending on size even at the same wavelength. This implies that using the UV absorption-based calibration may cause significant errors in GO quantification. Ultimately, this leads to incorrect dosages and faulty conclusions. This may also explain a variety of inconsistent results obtained in previous biological applications of GO. Graphical abstract The size of graphene oxide (GO) determines its UV absorption and the UV absorption-based calibration (GO-s, GO-m and GO-l represent the GO with small, medium and large size).

Study on the diagnosis of gout with xanthine and hypoxanthine.

BACKGROUND: Hyperuricemia is the only biochemical index in the classification of acute gouty arthritis in American Rheumatism Association 1977 and the main basis of clinical diagnosis for most doctors. However, nearly half of the time gout occurs without hyperuricemia, especially in an acute attack，which leads to an urgent need to find a new substitute diadynamic criteria of gout. Xanthine and hypoxanthine, as precursors of uric acid, have been reported to be high in gout patients with hyperuricemia and presumed to be gout biomarkers. OBJECTIVES: To further explore the possibility of xanthine and hypoxanthine to be gout biomarkers as substitutes for uric acid. METHODS: A reversed-phase HPLC-UV method was employed for simultaneous quantitative detection of uric acid (UA), xanthine (X), and hypoxanthine (HX) in gout patients' (with and without hyperuricemia) and healthy persons' serum. RESULTS: The xanthine and hypoxanthine concentrations in gout patients with hyperuricemia and without hyperuricemia are higher than in healthy persons with a P < 0.001. CONCLUSIONS: This study supplements previous researches by confirming that xanthine and hypoxanthine are significantly elevated in gout patients' serum especially in patients' with normouricemia, which supported xanthine and hypoxanthine may have clinical application for the diagnosis of gout.

Risk of Dementia in Long-Term Benzodiazepine Users: Evidence from a Meta-Analysis of Observational Studies.

BACKGROUND AND PURPOSE: There is conflicting evidence in the literature on the association between benzodiazepines (BDZs) and the risk of dementia. This meta-analysis aimed to determine the relationship between the long-term usage of BDZs and the risk of dementia. METHODS: The PubMed and Embase databases were systematically searched for relevant publications up to September 2017. The literature search focused on observational studies that analyzed the relationship between the long-term use of BDZs and the risk of dementia. Pooled rate ratios (RRs) with 95% confidence interval (CI) were assessed using a random-effects model. The robustness of the results was checked by performing subgroup and sensitivity analyses. RESULTS: Ten studies were included: six case-control and four cohort studies. The pooled RR for developing dementia was 1.51 (95% CI=1.17-1.95, p=0.002) in patients taking BDZ. The risk of dementia was higher in patients taking BDZs with a longer half-life (RR=1.16, 95% CI=0.95-1.41, p=0.150) and for a longer time (RR=1.21, 95% CI=1.04-1.40, p=0.016). CONCLUSIONS: This meta-analysis that pooled ten studies has shown that BDZ significantly increases the risk of dementia in the elderly population. The risk is higher in patients taking BDZ with a longer half-life (>20 hours) and for a longer duration (>3 years).

Chaiqin Chengqi decoction inhibits inflammatory mediators and attenuates acute pancreatitis through deactivation of janus kinase/signal transducer and activator of transcription signaling pathway.

OBJECTIVE: To investigate the effect of Chaiqin Chengqi decoction (CQCQD) on acute pancreatitis (AP) by janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in vitro and in vivo. METHODS: AP was induced by caerulein both in AR42J cells and in mice. AR42J cells were divided into five groups: the control group, the AP group, the CQCQD group, JAK/STAT signaling pathway inhibitor AG490 group, and the CQCQD and AG490 group. After induction, cellular supernatant of five groups were collected for measuring the concentrations of inflammatory cytokine amylase, interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), nuclear factor κB (NF-κB) by enzyme-linked immunosorbent assay and the expression of JAK-2, STAT-3 signaling transduction proteins by Western blot, respectively. Experiments in mice were performed similar to that of in AR42J cells. RESULTS: Treatment of AR42J cells with CQCQD reduced the pancreatic injury and negatively regulated the activities of amylase, as well as inhibited expression of several inflammatory cytokines such as IL-6, TNF-α, IL-1ß, NF-κB. Administration of CQCQD significantly inhibited JAK-2 activation and down-regulated phosphorylation of downstream substrate STAT-3 the same as AG490, resulting in inhibition of inflammatory mediators and amelioration of pancreatitis. CONCLUSION: The results suggested that CQCQD exerted anti-inflammatory effects on AP via reducing expression and phosphorylation of JAK and STAT.

Antioxidant and immunomodulatory activities of a polysaccharide from Flammulina velutipes.

OBJECTIVE: To evaluate the antioxidant and immunomodulatory activities of a unique polysaccharide from the medicinal fungus Flammulina velutipes in vitro. METHODS: Using water extraction and alcohol precipitation, crude polysaccharides were obtained. After purification by DEAE-cellulose 52 ion exchange chromatography and Sephacryl S-300 HR gel filtration chromatography, High performance liquid chromatography equipped with evaporative light-scattering detector, Infrared radiation and Nuclear magnetic resonance were used to evaluate the structure of the polysaccharide. Its immunomodulatory activity was measured by examining the production of nitric oxide (NO) and cytokine secretion, and via lymphocyte proliferation experiments. Its effects on the scavenging activities of hydroxyl radical, superoxide anion and reducing power were also measured. RESULTS: A water-soluble polysaccharide, Flammulina velutipes polysaccharide I-A (FVP I-A), was obtained with a molecular mass of 8.14 x 10(4) Da determined by high performance gel permeation chromatography. An in vitro antioxidant assay indicated that FVP I-A could scavenge hydroxyl radical, superoxide anion and possessed reducing power and could largely promote NO production and augment the interleukin-1ß, interleukin-6, and tumor necrosis factor-α secretion by RAW264.7 macrophages (P < 0.05). Moreover, FVP I-A could promote lymphocyte proliferation (P < 0.05), and synergistically enhance the augmentation of the proliferation of mouse lymphocytes by concanavalin A and lipopolysaccharides (P < 0.01, P < 0.05). CONCLUSION: The FVP I-A obtained from Flammulina velutipes possessed antioxidant activity and could enhance non-specific and specific immune responses in vitro.

[Effects of urokinase intervention on endotoxin-induced DIC in Wistar rat model].

OBJECTIVE: To study the effects of urokinase intervention on endotoxin-induced DIC in Wistar rats model. METHODS: Wistar rats were randomly assigned into 4 groups: normal saline (NS) group, urokinase (UK) group, endotoxin (LPS) group and LPS+ UK group. These agents were given to the rats by the tail vein intravenous infusion, NS group was treated with NS 2.5 mL/h x 4 h; UK group with NS 2.5 mL/h, 1 h later UK 4 IU/(g x h) x 3 h; LPS group with LPS 3 mg/(kg x h) x 4 h; LPS+UK group with LPS 3 mg/(kg x h) firstly, 1 h later UK 4 IU/(g x h) ) < 3 h. After the intervention, the function of coagulation and fibrinolysis, the indicators of organ damage and microcirculation fibrin micro-clots were evaluated. RESULTS: One hour after the infusion of 3 mg/(kg x h) of LPS, DIC signs began to appear, and became more apparent over time. After the intervention of urokinase, the values of clotting time (PT), activated partial thromboplastin time (APTT) were significantly shorter, but the platelet count (PLT), the amount of fibrinogen (FIB) changed little. Plasminogen activator inhibitor-1 (PAI-1) level decreased, while the D-dimer level increased. Serum creatinine (Cr), alanine aminotransferase (ALT) also decreased significantly. The biopsy of liver, kidney, and lung showed tissue damage became better, and the organ microcirculation fibrin micro-clots decreased significantly. CONCLUSION: The concentration of 3 mg/(kg x h) endotoxin can successfully induce DIC model in Wistar rats. Urokinase could play a positive role to prevent the LPS-induced DIC.

An assessment of the genetic diversity within Ganoderma strains with AFLP and ITS PCR-RFLP.

Ganoderma lucidum is one of the most important medicinal materials and plant pathogens. Because of its specific interhybridization, the genetic background, however, is relatively unclear. It made identification of Ganoderma strains, especially closely related strains difficulty. Amplified fragment length polymorphism (AFLP) using 14 primer combinations and internal transcribed spacer (ITS) PCR-RFLP were used in a comparative study which was designed to investigate the closely related Ganoderma strains genetic relations at molecular level. The analysis of 37 Ganoderma strains showed there were 177 polymorphic AFLP markers and 12 ITS PCR-RFLP markers, and all accessions could be uniquely identified. Among the Ganoderma accessions, similarity coefficients ranged from 0.07692 to 0.99194 in AFLP. The Ganoderma strains formed a tight cluster in nine groups in AFLP whereas seven groups in ITS PCR-RFLP. The cluster analysis revealed that the taxonomical system of subgenus Ganoderma is composed of Sect. Ganoderma and Sect. Phaeonema, and the strain 22 should be a variant form of strain 21. All methods delineated the Ganoderma strains from the different regions seeming to show a greater level of genetic diversity. It indicated that the genotype study at molecular level is a useful complement method to the current classification system of Ganoderma strains based on morphological traits. The congruency of the experiments was analyzed using the biostatistical software DPS V3.01.