Enhancing oncolytic virus efficiency with methionine and N-(3-aminoprolil)methacrylamide modified acrylamide cationic block polymer.

Oncolytic virus ablation of tumor cells has the advantages of high tumor selectivity, strong immunogenicity, and low side effects. However, the recognition and clearance of oncolytic viruses by the immune system are the main factors limiting their anti-tumor efficiency. As a highly biosafe and highly modifiable oncolytic virus vector, acrylamide can improve the long-term circulation of oncolytic viruses. Still, it is limited in its uptake efficiency by tumor cells. Herein, we constructed an N-hydroxymethyl acrylamide-b-(N-3-aminopropyl methacrylamide)-b-DMC block copolymer (NMA-b-APMA-b-DMA, NAD) as an oncolytic virus carrier, which not only improves the long-term circulation of oncolytic viruses in the body but also shows excellent stability for loading an oncolytic virus. The data shows that there was no obvious difference in the transfection effect of the NAD/Ad complex with or without neutralizing antibodies in the medium, which meant that the cationic carrier mediated by NAD/Ad had good serum stability. Only 10 micrograms of NAD carrier are needed to load the oncolytic virus, which can increase the transfection efficiency by 50 times. Cell experiments and mouse animal experiments show that NAD vectors can significantly enhance the anti-tumor effect of oncolytic viruses. We hope that this work will promote the application of acrylamide as an oncolytic virus vector and provide new ideas for methods to modify acrylamide for biomedical applications.

Camptothecin enhances the anti-tumor effect of low-dose apatinib combined with PD-1 inhibitor on hepatocellular carcinoma.

Apatinib has been shown to apply to a variety of solid tumors, including advanced hepatocellular carcinoma. Preclinical and preliminary clinical results confirmed the synergistic antitumor effects of apatinib in combination with anti-programmed death-1 (PD-1) inhibitors. In this study, we investigated camptothecin (CPT) enhances the anti-tumor effect of low-dose apatinib combined with PD-1 inhibitor on hepatocellular carcinoma. CPT combined with a PD-1 inhibitor enhances the anti-tumor effects of low-dose apatinib in hepatocellular carcinoma which was evaluated in making use of the H22 mouse model (n = 32), which was divided into four groups. Immunohistochemical staining and western blotting were used to detect nuclear factor erythroid 2-related factor 2 (Nrf2) as well as sequestosome 1 (p62), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2), PD-1, and programmed cell death ligand 1 (PD-L1). The results showed that the average size of the tumor of the combination group (Group D) was significantly less than that of the apatinib + PD-1 inhibitor group (Group C). The expression levels of Nrf2, p62, VEGFA, VEGFR2, PD-1, and PD-L1 in the apatinib + PD-1 inhibitor group(Group C) were lower than those in the control group (Group A) (P < 0.05). The expression levels of these genes in the apatinib + PD-1 inhibitor group (Group C) were significantly lower in the combination group (Group D) (P < 0.05). There was no obvious difference in body weight and liver and kidney functions between the four groups of mice. In conclusion, CPT improves the anti-tumor effect of low-dose apatinib combined with PD-1 inhibitor on hepatocellular carcinoma.

Tumor markers are associated with rapidly progressive interstitial lung disease in adult-dermatomyositis.

OBJECTIVES: DM-associated rapidly progressive interstitial lung disease (DM-RP-ILD) has been the clinical conundrum. We assess the serum levels of tumor markers (TMs) in different types of ILD, and explore the diagnostic utility of TMs for DM/ADM-RP-ILD. METHODS: This was a retrospective cohort study, data including clinical and laboratory records were collected from the first affiliated hospital of Zhengzhou University from December 2015, to June 2020. Tumor markers (TMs) include CEA, CA153, CA724, CA125, and CA199. Spearman analysis, ROC, and Kaplan-Meier curve were used for data analysis. RESULTS: Total 272 patients (149 DM and 123 ADM) were enrolled, 152 (55.88%) with ILD (116 with chronic ILD, 36 with RP-ILD) and 120 (44.12%) without ILD among them. The serum levels of CEA and ferritin were significantly higher in patients with RP-ILD than in the other two groups. Serum CA125, CA199, and CA153 levels in patients with RP-ILD were higher than those without ILD. CEA levels were associated with the ferritin, KL-6 and anti-MDA5 levels, and CEA concentration was significantly negatively correlated with DLco (P = 0.016, R2 = - 0.281). CEA [AUC = 0.7, 95% CI = (0.594, 0.806)] and ferritin [AUC = 0.737, 95% CI = (0.614, 0.860)] had diagnosed value for patients developing RP-ILD. Patients with high serum CEA levels had higher mortality rate within the DM-ILD population. CONCLUSIONS: TMs and ferritin were increased in DM/ADM-RP-ILD, and serum CEA and CA153 levels can evaluate disease severity of DM. And CEA and ferritin can be used as noninvasive diagnostic biomarkers for patients with DM-RP-ILD. Key Points • Interstitial lung disease (ILD) is a serious complication of DM, and is a leading cause of mortality, especially rapidly progressive ILD. • Tumor markers as a kind of noninvasive detection can reflect the disease severity of DM, and CEA and ferritin can be used to identify patients with RP-ILD.

[Deaths and life expectancy losses attributed to high-salt diet in Shandong province].

Objective: To estimate the deaths and life expectancy losses attributed to diet with high salt in Shandong province. Methods: Based on 24 h urinary sodium and blood pressure levels from the final evaluation survey of Shandong-Ministry of Health Action on Salt and Hypertension Project (SMASH) in 2016 and death cause data from Shandong Mortality Surveillance System, the population attributable fractions (PAF) and the deaths due to high-salt diet were calculated based on the framework of comparative risk assessment and the life expectancy loss was calculated by life table method. Results: A total of 32 987 deaths caused by high-salt diets were reported in 2016, accounting for 11.74% of related disease deaths and 4.95% of all deaths. The proportion of deaths due to high-salt diet in men (13.51%) was higher than that in women (9.17%). Cardiovascular diseases were the major causes of deaths due to high salt diet, accounting for 90.82% of all disease deaths caused by high-salt diets. The other causes were gastric cancer (8.10%) and chronic kidney disease (1.08%). The PAF in urban residents (13.87%) was higher than that in rural residents (10.87%). A loss of 0.58 years of the life expectancy were attributed to the high-salt diet. The different diseases caused by high-salt diet had different effects on life expectancy loss, ischemic heart disease ranked first, followed by cerebral hemorrhage and cerebral infarction. Conclusions: The proportion of deaths attributed to high-salt diets was high in Shandong. Cardiovascular diseases were the most important causes of deaths caused by high-salt diets. High-salt diet is still seriously affecting the health of residents in Shandong, indicating that salt reduction interventions need to be strengthened.

Behcet's disease with peripheral nervous system involvement successfully treated with golimumab: a case report and review of the literature.

Nervous system involvement is a rare and serious complication of Behcet's disease (BD), and the peripheral type is rarer. This article aimed to describe a case of BD with the peripheral nervous system (PNS) involvement and present a comprehensive literature review. One case of BD with PNS involvement was reported and related literature was retrospectively reviewed via PubMed/MEDLINE and Scopus database. The patient was resistant to traditional treatments, such as glucocorticoids and immunosuppressants, but had rapid quiescence after using golimumab. Our literature review suggests that the involved peripheral nerves in BD were diverse, the most common were the tibial nerves and peroneal nerves, vasculitis might be the main cause, and prednisone was still the cornerstone of treatment. TNF-α inhibitors have been increasingly used for refractory BD in recent years. This well-illustrated case demonstrates the potential benefit of golimumab to the patient with PNS involvement. Given the diversity and complexity of PNS involvement, we recommend golimumab as a new trial treatment in clinical practice.

Co-fermentation of lentils using lactic acid bacteria and Bacillus subtilis natto increases functional and antioxidant components.

We identified lentil products with both nutritional value and antioxidant capacity by studying the changes of probiotics and functional substances during single fermentation with lactic acid bacteria (LAB) or co-fermentation using LAB and Bacillus subtilis natto. After fermentation, the best growth of LAB was observed in anaerobic solid-state co-fermentation, whereby the viable counts of Lactobacillus plantarum TK9 and Lactobacillus paracasei TK1501 reached 2.77 × 109 and 2.78 × 109 CFU/g, respectively. Furthermore, the total phenol and genistin content produced by the two mixed groups, respectively, increased by 0.52- and 0.66-fold, as well as 0.63- and 0.64-fold, compared with unfermented samples. Similarly, the free amino acid content increased by 0.53- and 0.49-fold, respectively. The 50% inhibitory concentrations for the radical-scavenging against 1,1-diphenyl-2-picrylhydrazyl and α-glucosidase inhibitory activity were lower following anaerobic co-fermentation. Consistently, products of anaerobic mixed solid-state fermentation had higher oxygen radical absorbance capacity. Therefore, anaerobic solid-state co-fermentation of lentils using B. subtilis natto may promote the multiplication of LAB and enhance the antioxidant activity of fermented lentil products. PRACTICAL APPLICATION: Simple and efficient food handling is more suitable for industrial production. Co-fermentation is a good method to optimize the fermentation process. Co-culture technology has high potential in terms of functionality and antioxidant capacity.

Effects of ozone concentration on the postharvest quality and microbial diversity of Muscat Hamburg grapes.

Grapevines are widely planted around the world. Although grapes have high nutritional value, they are highly perishable. To explore the effect of ozone concentration on the postharvest quality of Muscat Hamburg grapes, the ethylene production rate, respiratory intensity, soluble solids, titratable acidity, firmness, threshing rate, total yeast and mold counts, and the activities of superoxide dismutase, peroxidase, catalase, polyphenol oxidase and phenylalanine ammonia lyase were determined, and the fungal metagenome on the grape surface was analyzed. Among the ozone treatment groups, 14.98 mg m-3 ozone showed a positive effect on grape preservation. After 80 days of storage, the contents of soluble solids and titratable acidity increased by 3.1% and 0.03%, respectively, compared with the control group. Over the same period, firmness increased by 4.22 N and the threshing rate decreased by 0.5%. During storage, the activity of polyphenol oxidase was inhibited and the activities of superoxide dismutase, peroxidase, catalase, and phenylalanine ammonia lyase were maintained, which delayed the senescence of grapes and maintained freshness. Ozone can reduce the number of fungi on the grape surface, change the colony structure, and reduce the occurrence of diseases. An ozone concentration of 14.98 mg m-3 can delay the senescence of Muscat Hamburg grapes and improve storage quality.

A Multi-crosslinking Nanocapsule-Based Serial-Stimuli-Responsive Leakage-Free Drug-Delivery System In Vitro.

As some stimuli utilized in conventional drug delivery systems can also be found in normal cells, it is inevitable that encapsulated drugs escape from carriers into normal cells. Based on mutual interactions among proteins, polyphenol compounds, and metal ions, we developed a serial-stimuli-responsive drug delivery system. With multi-crosslinking structure, nanocapsules can maintain the integrity of the framework, even with a certain amount of stimuli present, and eventually reach tumor cells to initiate apoptosis, and protect normal cells from being damaged. Meanwhile, the fluorescence of DOX will be quenched when encapsulated in nanocapsules. This property means that the DOX that is released from nanocapsules can be monitored in real-time based on the recovery of fluorescence. These versatile nanocapsules exhibit great potentials to treat cancer.

A sensitive ratiometric fluorescent probe for quantitive detection and imaging of alkaline phosphatase in living cells.

We report a sensitive fluorescence probe Ratio-ALP for in vivo ratiometric imaging of alkaline phosphatase (ALP). The probe was designed by conjugating a naphthalimide with a phosphate group as the responsive moiety for enzymatic reactions. The fluorescence intensity ratio displayed a linear relationship against the concentration of ALP in the concentration range from 0 to 200 U/L, its LOD is as low as 0.25 U/L. Ratio-ALP has good selectivity and sensitivity, high cell permeability, and low cytotoxicity. Furthermore, probe Ratio-ALP was also successfully applied to image endogenous ALP activity in living cells.

Dual-Site Fluorescent Probe to Monitor Intracellular Nitroxyl and GSH-GSSG Oscillations.

Nitroxyl (HNO), the one-electron-reduction product of NO has recently been revealed to have potentially beneficial pharmacological properties in cardiovascular health as a result of interactions with specific thiols such as glutathione (GSH). To disentangle the complicated inter-relationship between HNO and GSH in the signal transduction and oxidative pathways, we designed and synthesized a dual-site fluorescent probe NCF to indicate cellular HNO and GSH-GSSG balance. The sensitive and selective detection of HNO was achieved by incorporating an organophosphine group to naphthaldehyde-TCF. Then the resulted fluorescent product is able to monitor the conversion of GSH and GSSG reversibly. Additionally, outstanding biocompatibility make it capable of monitoring intracellular HNO and consequently GSH-GSSG oscillationsin living cells. We anticipate that NCF will be a unique molecular tool to investigate the interplaying roles of HNO and GSH.

Vitamin D protects podocytes from autoantibodies induced injury in lupus nephritis by reducing aberrant autophagy.

SUBJECT: The aim of this study was to investigate whether vitamin D plays a protective role in podocyte injury induced by autoantibodies purified from the serum of patients with lupus nephritis (LN) via reducing aberrant autophagy. METHODS: Autophagic activities of renal tissues of patients with LN were evaluated under transmission electronic microscope (TEM). Immunoglobulin G (IgG) from patients with LN was purified to induce human podocyte injury, and the role of vitamin D in injury was observed. Podocytes were observed under TEM, autophagic activity was evaluated by western blot analysis and quantitative real-time polymerase chain reaction, and mRFP-GFP-LC3B adenovirus was infected into human podocytes in vitro. RESULTS: Significantly higher autophagic levels were observed in patients with LN (P <0.05), and apparently greater autophagic levels in podocytes were shown (P <0.05). Among different classifications of LN, class V (n = 5), III + V (n = 5), and IV + V (n = 5) gained higher autophagic levels than class III (n = 5) and IV (n = 5). Induced autophagy, which was evident by increased LC3B-II and Beclin 1 level, caused consumption of p62, more autophagosomes observed under TEM, and more LC3B dots observed under confocal microscope in the IgG group, along with decreased nephrin expression, which suggests podocyte injury. Reduction of autophagy as well as alleviated podocyte injury was observed in the IgG+ vitamin D group. CONCLUSION: This study demonstrates that vitamin D plays a protective role in podocyte injury induced by autoantibodies from patients with LN and appears to be a novel therapy target in LN.

A two-photon fluorescent probe for basal formaldehyde imaging in zebrafish and visualization of mitochondrial damage induced by FA stress.

The detection of mitochondrial formaldehyde (FA) is of great significance because FA is generated through a one-carbon formaldehyde cycle in mitochondria, and abnormal elevations in FA levels can damage mitochondria by decreasing the mitochondrial membrane potential and inhibiting mitochondrial respiration. Herein, a mitochondria-targetable two-photon probe (Mito-FA-FP) has been well demonstrated. Mito-FA-FP is conjugated with hydrazine as the FA-reactive site and a pyridine derivate as the mitochondria-targetable moiety. After reacting with FA, Mito-FA-FP exhibits dramatic fluorescence enhancement (12-fold) due to suppression of the PET process in the probe and presents good selectivity as well as high sensitivity (LOD: 12.4 µM). Moreover, Mito-FA-FP can be utilized to monitor endogenous FA in mitochondria and evaluate mitochondrial damage caused by FA stress through observing mitochondrial morphology changes. With good two-photon absorption cross-section and high two-photon fluorescence contrast, Mito-FA-FP has been successfully employed for the two-photon fluorescence imaging of basal FA in zebrafish.

Specific and sensitive imaging of basal cysteine over homocysteine in living cells.

Biological thiols play important roles in maintaining appropriate redox status of organisms. Accepting the challenge to differentiate structurally similar cysteine (Cys) and homocysteine (Hcy), we have successfully developed a miniature synthetic turn-on fluorescent probe based on 6-(2-benzothiazolyl)-2-naphthalenol for Cys. This probe is able to specifically react with Cys to yield its naphthalenol derivative, accompanied by remarkable green fluorescence enhancement with a detection limit of 14.8 nM. Besides, this probe displays much greater selectivity for Cys over other biological thiols, including homocysteine (Hcy) and glutathione (GSH). Practically, good cell permeability and low cytotoxicity make it suitable for monitoring basal Cys in living cells.

Ratiometric fluorescence imaging of endogenous selenocysteine in cancer cell matrix.

Monitoring intracellular selenocysteine (Sec) is of significant interest for studying Sec metabolism and disease-relevant changes in Sec homeostasis. Herein, we rationally designed an ICT-based ratiometric probe (Rat-Sec) for selectively discriminating endogenous Sec in a live cell matrix. By utilizing an acrylate moiety, Rat-Sec manifested significantly ratiometric and red-shifted (∼117 nm) fluorescence signals within 1 min, and concomitantly demonstrated apparent color alteration from colorless to faint yellow in the presence of Sec. When applied to quantitatively detect Sec under physiological conditions, Rat-Sec gives a detection limit of 12 nM. Offering great advantages due to its ratiometric nature and high specificity and selectivity, Rat-Sec provides reliable fluorescence quantification and rapid tracking of exogenous and endogenous Sec in living cells.