The two-component system expression patterns and immune regulatory mechanism of Vibrio parahaemolyticus with different genotypes at the early stage of infection in THP-1 cells.

Vibrio parahaemolyticus must endure various challenging circumstances while being swallowed by phagocytes of the innate immune system. Moreover, bacteria should recognize and react to environmental signals quickly in host cells. Two-component system (TCS) is an important way for bacteria to perceive external environmental signals and transmit them to the interior to trigger the associated regulatory mechanism. However, the regulatory function of V. parahaemolyticus TCS in innate immune cells is unclear. Here, the expression patterns of TCS in V. parahaemolyticus-infected THP-1 cell-derived macrophages at the early stage were studied for the first time. Based on protein-protein interaction network analysis, we mined and analyzed seven critical TCS genes with excellent research value in the V. parahaemolyticus regulating macrophages, as shown below. VP1503, VP1502, VPA0021, and VPA0182 could regulate the ATP-binding-cassette (ABC) transport system. VP1735, uvrY, and peuR might interact with thermostable hemolysin proteins, DNA cleavage-related proteins, and TonB-dependent siderophore enterobactin receptor, respectively, which may assist V. parahaemolyticus in infected macrophages. Subsequently, the potential immune escape pathways of V. parahaemolyticus regulating macrophages were explored by RNA-seq. The results showed that V. parahaemolyticus might infect macrophages by controlling apoptosis, actin cytoskeleton, and cytokines. In addition, we found that the TCS (peuS/R) could enhance the toxicity of V. parahaemolyticus to macrophages and might contribute to the activation of macrophage apoptosis. IMPORTANCE This study could offer crucial new insights into the pathogenicity of V. parahaemolyticus without tdh and trh genes. In addition, we also provided a novel direction of inquiry into the pathogenic mechanism of V. parahaemolyticus and suggested several TCS key genes that may assist V. parahaemolyticus in innate immune regulation and interaction.

Composition engineering of AZO films for controlled photon-electron conversion and ultrafast nonlinear optical behavior.

Exploring micro-nano photonic crystals as nonlinear optical switching and optical limiting devices for Gaussian light fields with ultrashort pulse widths has attracted extensive research, mainly originating from its controllable modulation of the third/fifth-order nonlinear optical behavior and ultrafast carrier dynamics. In this work, Al-doped ZnO (AZO) films with controllable and excellent third-order nonlinear optical behavior have been uniformly deposited on quartz substrates by a single-step co-sputtering method. Al dopant-dependent ultrafast carrier dynamics and nonlinear optical properties in hexagonal ZnO films are discussed. The bonding mode of Al atoms in the ZnO lattice changed from substitutional to substitutional-decoration, which has been controllably achieved at different DC sputtering powers. The strain, crystallinity, grain size, dislocation density, and texture coefficient of the sample were quantitatively calculated by XRD and Raman spectroscopy, which confirmed that the phase parameters can be regulated by the sputtering power. In addition, Hall test and photoluminescence spectra showed the contribution of the donor level on the band structure and the electron transfer characteristics, which will provide a strategy for understanding multi-type carrier dynamics under strong light fields. The finite-difference time-domain method was used to simulate the linear optical absorption/transmittance of the sample under a plane-wave optical field, which proved that the light-matter interaction failed to be significantly suppressed by shading and scattering effects. The carrier relaxation process and nonlinear absorption/refractive effects were controllably optimized by dopant Al atoms, which were confirmed by Z-scan and transient absorption spectroscopy. Compared with pure ZnO films, the third-order nonlinear refraction and absorption coefficients of AZO-power films can reach -8.926 × 10-15 m2 W-1 and -0.634 × 10-7 m W-1, respectively. AZO films with ultrafast carrier dynamics and controllable excellent third-order nonlinear optical coefficients can be used as all-optical switches and optical limiting devices, which provide a reference for advanced micro-nano optical materials.

Fate of Vibrio parahaemolyticus on shrimp after acidic electrolyzed water treatment.

The objective of this study was to investigate the fate of Vibrio parahaemolyticus on shrimp after acidic electrolyzed water (AEW) treatment during storage. Shrimp, inoculated with a cocktail of four strains of V. parahaemolyticus, were stored at different temperatures (4-30 °C) after AEW treatment. Experimental data were fitted to modified Gompertz and Log-linear models. The fate of V. parahaemolyticus was determined based on the growth and survival kinetics parameters (lag time, λ; the maximum growth rate, µmax; the maximum growth concentration, D; the inactivation value, K) depending on the respective storage conditions. Moreover, real-time PCR was employed to study the population dynamics of this pathogen during the refrigeration temperature storage (10, 7, 4 °C). The results showed that AEW treatment could markedly (p<0.05) decrease the growth rate (µmax) and extend the lag time (λ) during the post-treatment storage at 30, 25, 20 and 15 °C, while it did not present a capability to lower the maximum growth concentration (D). AEW treatment increased the sensitivity of V. parahaemolyticus to refrigeration temperatures, indicated by a higher (p<0.05) inactivation value (K) of V. parahaemolyticus, especially for 10 °C storage. The results also revealed that AEW treatment could completely suppress the proliferation of V. parahaemolyticus in combination with refrigeration temperature. Based on above analysis, the present study demonstrates the potential of AEW in growth inhibition or death acceleration of V. parahaemolyticus on seafood, hence to greatly reduce the risk of illness caused by this pathogen during post-treatment storage.

Two novel thermostable chitinase genes from thermophilic fungi: cloning, expression and characterization.

Two chitinase genes, Tachit1 from Thermoascus aurantiacus var. levisporus and Ctchit1 from Chaetomium thermophilum were isolated. Tachit1 and Ctchit1 encode putative single-domain proteins (TaCHIT1 and CtCHIT1) of 399 and 402 amino acid residues, respectively. The catalytic domains of TaCHIT1 and CtCHIT1 are similar to those of other fungal chitinases in family 18 of glycosyl hydrolases. TaCHIT1 and CtCHIT1 have a molecular weight of about 48.4 and 47.3kDa, respectively when produced in recombinant Pichia pastoris. The enzymes exhibited optimum catalytic activity at pH 8.0 and 50 degrees C for TaCHIT1 and at pH 5.5 and 60 degrees C for CtCHIT1. TaCHIT1 retained 95.3% of its activity after 60 min at 50 degrees C. CtCHIT1 was stable at 50 degrees C and retained 96.7% of its activity after 60 min incubation at 60 degrees C. The TaCHIT1 and CtCHIT1 produced Glc-NAc2 as the major product, when colloidal chitin was used as the substrate. The enzyme could not hydrolyze pNp-(GlcNAc), but hydrolyzed colloidal chitin, powdery chitin and chitosan. These features make these proteins potentially useful for applications requiring chitin hydrolysis at elevated temperatures.

[Experimental study on the effects of interferon-gamma gene on Tenon's capsule fibroblasts in vitro].

OBJECTIVE: To investigate the effects of interferon-gamma (IFN-gamma) gene on Tenon's capsule fibroblasts in vitro. METHODS: Using lipofectAMINE, IFN-gamma gene was transferred to human Tenon's capsule fibroblasts with plasmid pcDNA3IFN-gamma in vitro. Cells treated with plasmid pcDNA3 without IFN-gamma gene were used as the controls. The transfected fibroblasts were screened by G418. The expression of IFN-gamma was determined by RT-PCR, immunohistochemistry and flow cytometry assay. The effects of IFN-gamma on the proliferation of Tenon's capsule fibroblasts were evaluated by flow cytometry and MTT. RESULTS: The Tenon's capsule fibroblasts transferred the IFN-gamma gene could express the IFN-gamma RNA and protein transiently. The proliferation of the fibroblasts transferred the IFN-gamma gene was inhibited. CONCLUSION: The proliferation of the Tenon's capsule fibroblasts in vitro can be inhibited by transferred IFN-gamma gene.