Purpose: Malignant melanoma (MM), the most lethal skin cancer, is highly invasive and metastatic. These qualities are related to not only genetic mutations in MM itself but also the interaction of MM cells with the immune system and microenvironment. This study aimed to construct a combined immunotherapy and gene therapy drug delivery system for the effective treatment of MM. Methods: Mature dendritic cell (mDC) exosomes (mDexos) with immune induction functions were used as carriers. BRAF siRNA (siBRAF) with the ability to silence mutated BRAF in MM was encapsulated in mDexos by electroporation to construct a biomimetic nanosystem for the codelivery of immunotherapy and gene therapy drugs (siBRAF-mDexos) to the MM microenvironment. Then, we investigated the nanosystem’s serum stability and biocompatibility, uptake efficiency in mouse melanoma cells (B16-F10 cells), cytotoxicity against B16-F10 cells and inhibitory effect on BRAF expression. Furthermore, we evaluated its antimelanoma activity and safety in vivo. Results: SiBRAF-mDexos were nanosized. Compared to siBRAF, siBRAF-mDexos displayed significantly increased serum stability, biocompatibility, uptake efficiency in B16-F10 cells, and cytotoxicity to B16-F10 melanoma cells; they also had a significantly greater inhibitory effect on BRAF expression and induced T-lymphocyte proliferation. Moreover, compared with siBRAF, siBRAF-mDexos showed significantly enhanced anti-MM activity and a high level of safety in vivo. Conclusion: The study suggests that the siBRAF-mDexo biomimetic drug codelivery system can be used to effectively treat MM, which provides a new strategy for combined gene therapy and immunotherapy for MM.

Colistin is being considered as “the last ditch” treatment in many infections caused by Gram-negative stains. However, colistin is becoming increasingly invalid in treating patients who are infected with colistin-resistant Escherichia coli (E. coli) and Klebsiella Pneumoniae (K. pneumoniae). To cope with the continuous emergence of colistin resistance, the development of new drugs and therapies is highly imminent. Herein, in this work, we surprisingly found that the combination of quercetin with colistin could efficiently and synergistically eradicate the colistin-resistant E. coli and K. pneumoniae, as confirmed by the synergy checkboard and time-kill assay. Mechanismly, the treatment of quercetin combined with colistin could significantly downregulate the expression of mcr-1 and mgrB that are responsible for colistin-resistance, synergistically enhancing the bacterial cell membrane damage efficacy of colistin. The colistin/quercetin combination was notably efficient in eradicating the colistin-resistant E. coli and K. pneumoniae both in vitro and in vivo. Therefore, our results may provide an efficient alternative pathway against colistin-resistant E. coli and K. pneumoniae infections.

The effect of shear displacement on the directivity of permeability in fractures is studied in this paper. The studied fracture surface has 3D self-affine fractal characteristics that are created using the modified successive random addition (SRA) method. Fluid flow through the fracture is simulated using the COMSOL Multiphysics code based on the finite element method (FEM) by changing the angle between the shear direction and macroscopic flow direction. The evolutions of the aperture distribution and flow paths with changes in shear displacement are investigated, and the change in the equivalent permeability is evaluated. The results show that the mean aperture and its deviation for rough fractures increase as the shear displacement increases, and this change is accompanied by an increase in void spaces and decreasing contact areas between the upper and lower fracture surfaces. The flow paths become more tortuous, and the channeling flow effect occurs during the shear process. The equivalent permeability of the fractures varies as the inclination between the shear direction and macroscopic flow direction changes. The permeability with the largest magnitude exists in the direction perpendicular to the shear direction, and the permeability with the smallest magnitude exists in the direction parallel to the shear direction. The equivalent permeability of the fractures at other inclinations varies between the smallest and greatest values. Notably, larger inclinations correspond to higher permeability magnitudes. The ratio of the directional permeability to the permeability in the direction parallel to the shear direction varies between 1.03 and 2.71. This ratio tends to decrease as the shear displacement and JRC increase, which indicates that the directivity of the permeability is more obvious for fractures with smaller JRCs and smaller shear displacement.

Diabetic peripheral neuropathy (DPN) is a chronic and serious microvascular complication of diabetes linked to redox imbalance. Sestrin2, a novel inducible stress protein, participates in glucose metabolic regulation and redox homeostasis. However, the association between serum Sestrin2 and DPN is unknown. To explore the association between serum Sestrin2 and DPN in patients with type 2 diabetes mellitus (T2DM). A total of 96 T2DM patients and 39 healthy volunteers, matched by age and sex, participated in this cross-sectional study. Clinical features and metabolic indices were identified. Serum Sestrin2 was measured by ELISA. The association between Sestrin2 and DPN was studied. Correlation and logistic regression analyses were used to evaluate the associations of different metabolic indices with Sestrin2 and DPN. The 96 patients with T2DM were divided into DPN (n = 47) and patients without DPN (n = 49). Serum Sestrin2 was significantly lower in healthy volunteers than in all T2DM patients combined [9.10 (5.41-13.53) ng/mL vs 12.75 (7.44-23.80) ng/mL, P < 0.01]. T2DM patients without DPN also had significantly higher levels of Sestrin2 than healthy volunteers [14.58 (7.93-26.62) ng/mL vs 9.10 (5.41-13.53) ng/mL, P < 0.01]. However, T2DM patients with DPN had lower cir culating Sestrin2 levels compared to T2DM patients without DPN [9.86 (6.72-21.71) ng/mL vs 14.58 (7.93-26.62) ng/mL, respectively, P < 0.01]. Bivariate correlation analysis revealed that serum Sestrin2 was positively correlated with body mass index (r = 0.672, P = 0.000), hemoglobin A1c (HbA1c) (r = 0.292, P = 0.000), serum creatinine (r = 0.206, P = 0.016), triglycerides (r = 0.731, P = 0.000), and fasting glucose (r = 0.202, P = 0.040), and negatively associated with estimated glomerular filtration rate (r = -0.230, P = 0.007). After adjustment for sex, age, HbA1c, and diabetes duration, multiple regression analysis revealed that Sestrin2 was independently correlated with body mass index and triglyceride levels (P = 0.000). Logistic regression analyses indicated that Sestrin2, diabetes duration, and high-density lipoprotein were strongly associated with DPN (odds ratio = 0.855, 1.411, and 0.041, respectively). Our results show Sestrin2 is decreased in T2DM patients with DNP. As lower Sestrin2 is independently associated with DPN, Sestrin2 may contribute to progression of DPN in T2DM patients.

In order to investigate the combined effect of contact area, aperture variation, and fracture connectivity on the fluid flow through a fractured medium, a series of flow simulations were implemented on two types of three-dimensional discrete fracture network (3D DFN) models constituting fractures having spatially variable apertures and parallel plates, respectively. The flow tortuosity within the 3D DFN models was examined by changing the density, aperture distribution, and closure of fractures. The results show that compared with the 3D DFN models constituting parallel plates, the model with variable apertures provides more pronounced 3D preferential flow pathways. At the individual fracture scale, the preferential flow pathways mostly converge within the void spaces of large aperture, and at the network scale, they are located in the most transmissive fractures within the connected networks. The permeability of 3D DFNs depends not only on the contact area and aperture variation within individual fractures but also on the fracture connectivity and the contact at fracture intersections within the fracture network. Increasing the fracture connectivity tends to enhance the permeability, while increasing the contact at fracture intersections would significantly reduce the permeability. A correlation between the equivalent permeability of 3D DFNs constituting fractures with spatially variable apertures and parallel plates is proposed incorporating the effect of network-scale topology. A tortuosity factor for 3D DFNs is defined based on the proposed model, and it can account for two competing effects when the model is upscaled from individual fracture to fracture network: the permeability reduction induced by contact obstacles at fracture intersections and permeability enhancement induced by increasing the fracture connectivity.