Synergetic interaction of capsid proteins for virus-like particles assembly of foot-and-mouth disease virus (serotype O) from the inclusion bodies.

Recombinant virus-like particles (VLP) with single capsid protein have been successfully produced through prokaryotic system, but for those with multiple capsid proteins such as the foot-and-mouth disease virus (FMDV), this approach is more challenging. In this study, in vitro assembly of FMDV VLP was investigated with its capsids VP1, VP2 and VP3 separately expressed as inclusion bodies. After extraction and solubilization, three capsids were purified in denatured state through a flow-through model, increasing its purity to 90%. VLP assembly for FMDV was observed after diluting the mixture of denatured capsids in the ration of 1: 1: 1, while no VLP appeared if the separately diluted and refolded capsids were co-incubated. This result suggests certain synergetic interactions exist among the three capsids, which are crucial for FMDV VLP assembly. Sodium chloride and capsid protein concentration both greatly affect the assembling efficiency. After purification through size exclusion chromatography, VLP with similar diameter and morphology as inactivated FMDV were obtained, which elicited high IgG titers and B cell activation when vaccinated in mouse. It could also induce specific humoral and cellular immune responses in splenocytes proliferative experiments. Our study demonstrated the feasibility of in vitro assembling FMDV VLP from inclusion bodies of VP1, VP2 and VP3 for the first time.

Genetic Profiles and Three-year Follow-up Study of Chinese Males With Congenital Hypogonadotropic Hypogonadism.

BACKGROUND: The correlation between long-term treatment outcomes with genotypes in congenital hypogonadotropic hypogonadism (CHH) males is rarely reported. AIM: To investigate the correlations among genotypes, phenotypes, and treatment outcomes for CHH male patients. METHODS: Whole exome sequencing was performed for 73 Chinese CHH males from one academic center. Patients self-selected one of the 4 treatments: pulsatile Gonadorelin pump (PGP), cyclical gonadotropins therapy (CGT), human menopausal gonadotropin monotherapy, or testosterone replacement treatment. Clinical assessments were performed every 3 months for 3 years. OUTCOMES: The pathogenicity of variants was determined. Baseline clinical features, spermatogenesis outcomes were analysed. RESULTS: 62 variants were identified in 51 patients (69.9%), 17 of which were novel. Among these mutations, variants on FGFR1, PROKR2, CHD7, ANOS1 and NSMF gene were 16.1%, 16.1%, 11.3%, 8.1% and 8.1% respectively. 11 patients followed the oligogenic pattern (21.6%). All CHD7 patients had hearing impairment or structural deformities of external/inner ear, and were diagnosed as CHARGE syndrome. 24.7% of CHH patients manifested with ear/hearing anomalies. KS patients had higher rates of cryptorchidism history and ear/hearing anomalies than normosmic CHH subjects. Male patients with PROKR2 mutations showed relatively better testicular development, less dental deformity when compared with FGFR1 mutations. About 30% normosmic patients defined by simple olfactory assessment showed olfactory nerve center (ONC) dysplasia under nasal sinus MRI examination. Among the CHH males treated with CGT or PGP, 70.2% reached spermatogenesis within 3 years of treatment. CLINICAL IMPLICATIONS: No direct correlation was observed between certain responsible genes and spermatogenic outcomes. When CHH patients were identified with CHD7 variants, ear/hearing evaluation should be carefully performed. The precise assessment of ONC development was advised for normosmic CHH subjects. STRENGTHS & LIMITATIONS: This study provided informative long-term treatment data of CHH male patients screened with whole exome sequencing. The limitations included small number of subgroups with multifaceted gene variants, clinical heterogeneity, and uncontrolled sperm-inducing treatment method. The seventeen novel mutations worth experimental validation in the future. CONCLUSION: The clinical severity is partially related with specific gene variants, and detailed individualized data and outcomes were provided. Ear/hearing anomalies were closely connected with CHD7 variants, and were common problems for CHH patients. Simple olfactory assessment underestimated the true olfactory deficit. L. Zhang, Y. Gao, Q. Du, et al. Genetic Profiles and Three-year Follow-up Study of Chinese Males With Congenital Hypogonadotropic Hypogonadism. J Sex Med 2021;18:1500-1510.

Effect of ceramic type and thickness on the masking ability of chairside machinable all-ceramic crowns.

OBJECTIVES: This study aimed to evaluate the influence of ceramic type and thickness on the masking ability and final aesthetic effects of chairside machinable all-ceramic crowns. METHODS: Six kinds from three types chairside machinable ceramic materials (IPS e.max CAD HT/MT/LT, IPS Empress LT, and VITA Suprinity HT/T) in shade A2 were fabricated to slice specimens into 1.0, 1.5, and 2.0 mm-thick sections (n=10). The color parameters of the specimens against black and white tiles and four resin substrates (A2, A4, B3, and C4 shade) were measured with a spectrophotometer. The translucency parameter (TP) was calculated using color parameters measured over standard white and black backgrounds. The color differences (ΔE) were calculated between there substrate shades (A4, B3, C4 ) and A2 shade (control group). Two-way analysis of variance (ANOVA) was performed on the TP values. The two va-riables were ceramic type and ceramic thickness. Three-way ANOVA was used to determine the effects of ceramic materials, ceramic thickness, and substrate shades on the ΔE values, followed by Tukey test for multiple comparisons (α=0.05). RESULTS: Ceramic type, ceramic thickness, and substrate shade significantly affected the ΔE values (P<0.001). The L* and b* values of the specimens increased with increasing ceramic thickness, except in substrate A2, whereas the ΔE values decreased. The color difference of all 1.0 mm-thick specimens or all specimens over the substrates C4 shade exceeded the clinically acceptable threshold (ΔE>3.3). CONCLUSIONS: The masking ability of chairside machinable all-ceramic crowns is influenced by ceramic type and thickness, and ceramic material. The thickness of ceramic less than 2.0 mm cannot mask the gray shade abutment.

Lipopolysaccharide modification enhances the inhibitory effect of clodronate liposomes on hepatic fibrosis by depletion of macrophages and hepatic stellate cells.

Hepatic fibrosis is a complex chronic liver disease in which both macrophages and hepatic stellate cells (HSCs) play important roles. Many studies have shown that clodronate liposomes (CLD-lipos) effectively deplete macrophages. However, no liposomes have been developed that target both HSCs and macrophages. This study aimed to evaluate the therapeutic efficacy of lipopolysaccharide-coupled clodronate liposomes (LPS-CLD-lipos) and the effects of liposomes size on hepatic fibrosis. Three rat models of hepatic fibrosis were established in vivo; diethylnitrosamine (DEN), bile duct ligation (BDL), and carbon tetrachloride (CCl4). Hematoxylin and eosin staining and serological liver function indices were used to analyze pathological liver damage. Masson's trichrome and Sirius red staining were used to evaluate the effect of liposomes on liver collagen fibers. The hydroxyproline content in liver tissues was determined. In vitro cell counting kit-8 (CCK-8) and immunofluorescence assays were used to further explore the effects of LPS modification and liposomes size on the killing of macrophages and HSCs. Both in vitro and in vivo experiments showed that 200 nm LPS-CLD-lipos significantly inhibited hepatic fibrosis and the abnormal deposition of collagen fibers in the liver and improved the related indicators of liver function. Further results showed that 200 nm LPS-CLD-lipos increased the clearance of macrophages and induced apoptosis of hepatic stellate cells, significantly. The present study demonstrated that 200 nm LPS-CLD-lipos could significantly inhibit hepatic fibrosis and improve liver function-related indices and this study may provide novel ideas and directions for hepatic fibrosis treatment.

Super absorbent glutaric anhydride-modified agar: Structure, properties, and application in biomaterial delivery.

Agar was modified with glutaric anhydride (GA) in this study to expand its application in food and medicine. Glutaric anhydride-modified agar (GAR) can maintain high gel strength (1247.4 g/cm2) and improved transparency (82.7 %). The esterified agar formed by GA further formed a cross-linking molecule structure by increasing the reaction temperature. Notably, excellent freeze-thaw stability (24.1 %) and swelling property (3116.6 %) of GAR indicated that the carboxyl-terminal of modified agar improves its affinity with water. Therefore, satisfactory water permeability and expansive stone enable agar films to achieve high water absorption. Furthermore, GAR films exhibit a specific absorption capacity of tetracycline hydrochloride in weak acid solution, thereby suggesting its potential application as a sustainable drug delivery carrier. Finally, the structure of the modified agar was analyzed to explain the mechanism of binding water. Cryo-scanning electron microscopy (SEM) depicted the porous structure of the agar gel responsible for swelling, drug loading, and release. Low-field NMR results showed that GA improves agar gel's binding and free water content. According to our research results, these GAR hydrogel membranes with excellent properties have the potential to be used as effective drug delivery materials.

In situ titanium phosphate formation on a titanium implant as ultrahigh bonding with nano-hydroxyapatite coating for rapid osseointegration.

Titanium (Ti) has been widely used as a dental implant material due to its excellent mechanical property and good biocompatibility. However, its poor biological activity severely limits its ability to bond with bony tissues. To ameliorate this situation, a preparation method of ultra-high bonding nano-hydroxyapatite (n-HA) coating on the Ti surface is urgently needed. Here, Ti phosphate/n-HA (TiP-Ca) composite coatings with ultra-high bonding were prepared by a two-step hydrothermal treatment. The TiP coating was first formed in situ on the pure Ti substrate and then n-HA crystals further grew on the TiP surface. The formation mechanism of composite coating and reasons for increased bonding strength were systematically investigated. The results show that the TiP-Ca coating remains stable and exhibits an ultra-high bonding strength with the Ti implant (up to 783.30 ± 207.46 N). An effective solution was designed to address the problems of easy peel off. Cell experiments showed that TiP-Ca could promote the adhesion of MC3T3-E1 and expression of OCN, Runx2, and ALP. In vivo evaluation further confirmed that the TiP-Ca composite coating significantly enhanced osseointegration. The designed coating shows great potential in clinical application of implants.

Effect of soybean protein isolate-pectin composite nanoparticles and hydroxypropyl methyl cellulose on the formation, stabilization and lipidolysis of food-grade emulsions.

The formation and stabilization mechanism as well as digestion characteristics of food-grade emulsions prepared by the SPNPs-HPMC mixed systems (a combination of soybean protein isolate-pectin composite nanoparticles (SPNPs) and hydroxypropyl methylcellulose (HPMC)) were investigated. Then, it was found that the SPNPs-HPMC mixed systems could not only enhance the stability of the emulsion, but also make it have a satisfactory lipidolys is efficiency. During the formation and stabilization of the emulsion, HPMC was adsorbed in the early stage of emulsion formation, while SPNPs needed a longer adsorption time. When the HPMC concentration was 0.25-0.5 wt%, HPMC and SPNPs co-adsorbed on the interface. When the HPMC concentration was 1-2 wt%, HPMC and SPNPs competed to adsorb on the interface, of which the adsorption HPMC was dominant. In vitro simulation of digestion, SPNPs were decomposed into substances with lower interfacial activity, and the structure and activity of HPMC were well maintained, which led them to reconstruct a new interface layer. Thus, the size distribution and surface area of the emulsion droplets were retained in a good state for the lipidolysis process. Therefore, the SPNPs-HPMC mixed systems could both enhance the stability of the emulsion and grant it a satisfactory lipidolysis efficiency.

Polystyrene microplastics induced female reproductive toxicity in mice.

Plastics have caused serious environmental pollution. In recent years, microplastics (MPs) have caused widespread concern about their potential toxicity on animals and humans, especially on organ and tissue deposition. However, there is little known about the reproductive toxic effects of MPs in female mammals. In this study, the reproductive toxicity of polystyrene MPs (PS-MPs) in female mice was evaluated after continued exposure for 35 days. Results showed that PS-MPs could accumulate in heart, liver, spleen, lung, kidney, brain, large intestine, small intestine, uterus, ovary and blood of exposed mice. Moreover, PS-MPs exposure increased the IL-6 level and decreased malondialdehyde (MDA) level in mouse ovaries. The results also showed that PS-MPs exposure decreased the first polar body extrusion rate and the survival rate of superovulated oocytes. Meanwhile, PS-MPs reduced the level of glutathione (GSH), mitochondrial membrane potential (MMP), endoplasmic reticulum calcium ([Ca2+]ER) and increased reactive oxygen species (ROS) in oocytes. In conclusion, our study illustrated that PS-MPs exposure induced the inflammation of ovaries and reduced the quality of oocytes in mice, which provided a basis for studying the reproductive toxic mechanism of PS-MPs in female mammals.

Doxorubicin-liposome combined with clodronate-liposome inhibits hepatocellular carcinoma through the depletion of macrophages and tumor cells.

Macrophages in the liver have capacities of capturing and phagocytosing nanocarriers. Macrophages also play an important role in the inflammatory microenvironment and in the tumorigenesis, development and progression of hepatocellular carcinoma (HCC). Several studies have shown that depletion of macrophages is a viable strategy for drug delivery and tumor microenvironment regulation. We prepared liposomes containing doxorubicin and clodronate using an ammonium sulfate gradient and thin film hydration method. The repressive therapeutic effects of liposomes were compared by intrasplenic injection at different stages of a primary HCC model induced by diethylnitrosamine (DEN) in rats. Doxorubicin-liposome (DOX-LIP) and clodronate-liposome (CL-LIP) about 180-200 nm were successfully prepared and characterized. We found that DOX-LIP combined with CL-LIP could effectively inhibit the occurrence and development of liver cancer without major organ damage and side effects. The combination of doxorubicin and clodronate liposomes notably decreased hepatic CD68 + macrophages, enriched DOX in plasma and accumulated it for a long time in the liver and spleen, thus improving the tumor microenvironment, inhibiting the activation of hepatic progenitor cells (HPCs) and promoting the apoptosis of tumor cells, and finally producing the inhibitory and therapeutic effects of HCC in rats. Results of this study were expected to provide a new prospect for the chemotherapy of HCC.

The effects of cellulose nanocrystal and cellulose nanofiber on the properties of pumpkin starch-based composite films.

Pumpkin starch (PS) was extracted from Cucurbita maxima and utilized to prepare films in combination with cellulose nanocrystal (CNC) and cellulose nanofiber (CNF), using a solvent casting strategy. The PS was characterized to contain 26.6% of amylose, exhibiting a "B"-type crystalline structure and high stability against thermal degradation. PS/CNF films showed better thermal stability than PS/CNC films, whereas the CNC was more effective than CNF for enhancing the tensile strength (TS) of the films. The nanocomposite films containing 1% CNC showed the highest TS of 30.32 MPa. Fourier transform infrared spectra revealed stronger hydrogen bonding in the PS/CNC films, likely contributing to the observed high mechanical strength. CNC and CNF both decreased the transparency of PS films, by 5.2% and 13.1%, respectively. Overall, the properties of PS composite films can be effectively modified by incorporating CNC and CNF, as PS/CNC films with high mechanical strength and PS/CNF films with good thermal stability. Our results indicate that PS is a suitable material for CNC/CNF composite film fabrication. These films are expected to be especially useful in food packaging applications.