Multifunctional bispecific nanovesicles targeting SLAMF7 trigger potent antitumor immunity.

The effectiveness of immune checkpoint inhibitor (ICI) therapy is hindered by the ineffective infiltration and functioning of cytotoxic T cells and the immunosuppressive tumor microenvironment (TME). Signaling lymphocytic activation molecule family member 7 (SLAMF7) is a pivotal co-stimulatory receptor thought to simultaneously trigger natural killer (NK)-cell, T-cell, and macrophage antitumor cytotoxicity. However, the potential of this collaborative immune stimulation in antitumor immunity for solid tumors is under-explored due to the exclusive expression of SLAMF7 by hematopoietic cells. Here, we report the development and characterization of multifunctional bispecific nanovesicles targeting SLAMF7 and Glypican-3-a hepatocellular carcinoma (HCC)-specific tumor antigen. We found that by effectively "decorating" the surface of solid tumors with SLAMF7, these nanovesicles directly induced potent and specific antitumor immunity and remodeled the immunosuppressive TME, sensitizing the tumors to programmed cell death protein 1 (PD-1) blockade. Our findings highlight the potential of SLAMF7-targeted multifunctional bispecific nanovesicles as an anticancer strategy with implications for designing next-generation targeted cancer therapies.

First-principles study of LiFePO4 modified by graphene and defective graphene oxide.

The energy stability and electronic structural of graphene and defective graphene oxide (GO) parallel to the surface of LiFePO4 (010) were theoretically investigated by using first-principles density functional theory calculations within the DFT + U framework. The calculated formation energy shows that GO coating on the surface of LiFePO4 (010) is energetically favorable and has higher bond strength compared to graphene. The calculation of the electronic structure indicates that the emergence of band in-gap states originates from graphene coating, with adsorbed O atoms contributing significantly above the Fermi level. Electron density difference indicate that GO stands on the LFP (010) surface through C-O and Fe-O bonds, rather than relying on van der Waals forces placed parallel to the LFP crystal, with the chemical bond at the LFP/GO interface (Fe-O-C) both anchoring the coated carbon layer and promoting electron conductivity at the interface. In addition, LFP/GO shows superior electrochemical performance, Atomic Populations suggests that the average Fe-O bonding on the surface of LiFePO4 (010) was clearly changed after graphene or GO coating, which led to the expansion of Li+ channels and favored the migration insertion and extraction of Li+.

Damage-Associated Molecular Patterns, a Class of Potential Psoriasis Drug Targets.

Psoriasis is a chronic skin disorder that involves both innate and adaptive immune responses in its pathogenesis. Local tissue damage is a hallmark feature of psoriasis and other autoimmune diseases. In psoriasis, damage-associated molecular patterns (DAMPs) released by damaged local tissue act as danger signals and trigger inflammatory responses by recruiting and activating immune cells. They also stimulate the release of pro-inflammatory cytokines and chemokines, which exacerbate the inflammatory response and contribute to disease progression. Recent studies have highlighted the role of DAMPs as key regulators of immune responses involved in the initiation and maintenance of psoriatic inflammation. This review summarizes the current understanding of the immune mechanism of psoriasis, focusing on several important DAMPs and their mechanisms of action. We also discussed the potential of DAMPs as diagnostic and therapeutic targets for psoriasis, offering new insights into the development of more effective treatments for this challenging skin disease.

Seawater pearl hydrolysate inhibits photoaging via decreasing oxidative stress, autophagy and apoptosis of Ultraviolet B-induced human skin keratinocytes.

BACKGROUND: Ultraviolet (UV) is the main reason to cause photoaging skin which not only hinders beauty, brings the patients with psychological burden, but also pathologically leads to the occurrence of tumors in skin. OBJECTIVE: This study goes into the inhibitory effect and mechanism of seawater pearl hydrolysate (SPH) to address human skin keratinocytes photoaging induced by UVB. METHODS: The photoaging model of Hacat cell was constructed by UVB irradiation, the levels of oxidative stress, apoptosis, aging, autophagy and autophagy-related protein and signal pathway expression were assessed to characterize the inhibitory effect and mechanism of SPH on photoaging Hacat cell. RESULTS: Seawater pearl hydrolysate significantly accelerated (p < 0.05) the activities of superoxide dismutase, catalase, and glutathione peroxidase, and markedly reduced (p < 0.05) the contents of reactive oxygen species (ROS), malondialdehyde, protein carbonyl compound and nitrosylated tyrosine protein, aging level, apoptosis rate in Hacat cell induced by 200 mJ cm-2 UVB after 24 and 48 h of culture; high dose SPH significantly raised (p < 0.05) relative expression level of p-Akt, p-mTOR proteins, and markedly decreased (p < 0.05) relative expression level of LC3II protein, p-AMPK, and autophagy level in Hacat cell induced by 200 mJ cm-2 UVB, or in combination with the intervention of PI3K inhibitor or AMPK overexpression after 48 h of culture. CONCLUSION: Seawater pearl hydrolysate can effectively inhibit 200 mJ cm-2 UVB-induced photoaging of Hacat cells. The mechanism indicates removing the excessive ROS through increasing the antioxidation of photoaging Hacat cells. Once redundant ROS is eliminated, SPH works to reduce AMPK, increase PI3K-Akt pathway expression, activate mTOR pathway to lowdown autophagy level, and as a result, inhibit apoptosis and aging in photoaging Hacat cells.

Sequential Targeting Hybrid Nanovesicles Composed of Chimeric Antigen Receptor T-Cell-Derived Exosomes and Liposomes for Enhanced Cancer Immunochemotherapy.

Paclitaxel (PTX)-based chemotherapy remains the main approach to treating lung cancer but systemic toxicity limits its use. As chimeric antigen receptor-T (CAR-T) cell-derived exosomes contain tumor-targeted CARs and cytotoxic granules (granzyme B and perforin), they are considered potential delivery vehicles for PTX. However, the low drug-loading capacity and hepatotropic properties of exosomes are obstacles to their application to extrahepatic cancer. Here, a hybrid nanovesicle named Lip-CExo@PTX was designed for immunochemotherapy of lung cancer by fusing exosomes derived from bispecific CAR-T cells targeting both mesothelin (MSLN) and programmed death ligand-1 (PD-L1) with lung-targeted liposomes. Due to the lung-targeting ability of the liposomes, over 95% of intravenously administered Lip-CExo@PTX accumulated in lung tissue. In addition, with the help of the anti-MSLN single-chain variable fragment (scFv), the PTX and cytotoxic granules inside Lip-CExo@PTX were further delivered into MSLN-positive tumors. Notably, the anti-PD-L1 scFv on Lip-CExo@PTX blocked PD-L1 on the tumors to avoid T cell exhaustion and promoted PTX-induced immunogenic cell death. Furthermore, Lip-CExo@PTX prolonged the survival time of tumor-bearing mice in a CT-26 metastatic lung cancer model. Therefore, Lip-CExo@PTX may deliver PTX to tumor cells through sequential targeted delivery and enhance the antitumor effects, providing a promising strategy for immunochemotherapy of lung cancer.

Functionalized Rhodium Nanoparticles as Antimicrobial Agents for Treatment of Drug-Resistant Skin and Soft Tissue Infections.

Skin and soft tissue infections (SSTIs) are among the most common bacterial infections reported in outpatients. Drug-resistant bacteria are the major cause of treatment failure and increased mortality rate in patients with SSTIs, posing significant challenges to human health. In this study, new-generation rhodium nanoplates (RhNPs) and glycol chitosan- and polydopamine-functionalized RhNPs (Rh@GCS) are developed for the treatment of drug-resistant SSTIs. RhNPs exhibited favorable antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Ag-resistant MRSA. The modified Rh@GCS exhibited enhanced antibacterial activity and can directly kill various drug-resistant bacteria by increasing the permeability of cell membranes, including gram-positive MRSA and gram-negative multidrug-resistant Escherichia coli (E.coli) and Pseudomonas aeruginosa (PA). Moreover, Rh@GCS effectively inhibited bacterial growth and promoted the healing of skin lesions in MRSA-induced SSTI mouse models. These results suggest that Rh@GCS is a promising nonantibiotic antimicrobial agent for the treatment of drug-resistant SSTIs.

Study on the Mechanism of Hydrolyzed Seawater Pearl Tablet in Treating Chronic Sleep Deprivation Mice Model.

BACKGROUND: Modern lifestyle increasingly deprives people from sleep to different degrees. Long-term sleep deprivation will facilitate body's pathological behaviors, such as lethargy, depression, and anorexia. OBJECTIVE: This study is an investigation into the mechanism of hydrolyzed seawater pearl tablet in treating chronic sleep deprivation mice model. METHODS: The chronic sleep deprivation model was established involving C57BL/6mice; the body weight, behavioral characteristics, hippocampal structure, oxidative stress, apoptosis-related protein expression, and intestinal bacteria in mice were assessed to characterise hydrolyzed seawater pearl tablet. RESULTS: Hydrolyzed seawater pearl tablet significantly accelerated body weight, open field test score, and sugar water preference rate (P < 0.05), alleviated the structural damage of hippocampus, reduced the content of MDA (P < 0.05), Bax protein expression, increased the content of GSH (P < 0.05), the activities of SOD, GSH-Px, and Bcl-2 protein expression in the hippocampus, increased the Escherichia coli, Bacteroides, Bifidobacterium and Lactobacillus (P < 0.05), which are beneficial bacteria in the intestine, in chronic sleep deprivation mice, and reduced the amount of Clostridium perfringens (P < 0.05), which are harmful bacteria in the intestine. CONCLUSION: Hydrolyzed seawater pearl tablet can improve the depression-like mental state of mice caused by chronic sleep deprivation. The mechanism involves improving the antioxidant activity of the hippocampus to eliminate the excessive ROS, which inhibits cell apoptosis and alleviates tissue structure damage. Meanwhile, it may also be involved in adjusting the microbiota level and improving the mental and behavioral activities of chronic sleep deprivation mice through the intestine-brain axis.

Immune Effects of Macrophages in Rheumatoid Arthritis: A Bibliometric Analysis From 2000 to 2021.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease characterized by macrophage activation. The current characteristics, hotspots, and research frontiers of macrophage-related RA were analyzed using bibliometric analysis. Relatedpapers published from 2000 to 2021 in the Web of Science database were retrieved. The diagrams were generated and analyzed using the bibliometric software package. VOSviewer and CiteSpace were used to evaluate and visualize the research trends and hotspots in macrophage-related RA. A total of 7253 original articles were obtained. Global research on macrophage-related RA is in an advanced stage of development, with core authors, teams and research institutions emerging. United States has published the most papers, received the most citations, and had the highest H-index over the last 22 years. The University of Amsterdam and the journal of Arthritis and Rheumatism are the most productive research institutions and journals. Tak PP's (St Vincent's Hospital) paper has the highest publication and citation scores. The keywords "bone loss" and "polarization" have the highest frequency. Additionally, the study of macrophage polarization in RA has been research focus in recent years. This study demonstrates that research on macrophages in RA will continue. China is a significant producer, whereas the United States is an influential nation in this regard. In the last decade, most studies have concentrated on fundamental research. Recent studies have shown how macrophages play a role in controlling and weakening inflammation, and drug delivery and mechanism have come to the fore.

Inhibitory Effect of Seawater Pearl Hydrolysate on UVA-Induced Photoaging of Human Skin Fibroblasts.

This study is an investigation into the inhibitory effect of seawater pearl hydrolysate (SPH) on the UVA-induced photoaging of human skin fibroblast (HSF) cells, and the mechanism thereof. HSF cells were cultured and irradiated with a UVA 0-50 J·cm-2 dose gradient. The cell inhibition rate was detected using the CCK8 method, and the half-inhibitory dose was determined. Based on this, the dose of UVA irradiation for the follow-up experiment was selected to establish a photoaging model of the HSF cells. The cells were divided into a normal (N) group, UVA-irradiated (UVA) group, SPH low dose (SPHL) group, SPH medium dose (SPHM) group, and SPH high dose (SPHH) group. The photoaging model of HSF cells was established by UVA irradiation in the UVA, SPHL, SPHM, and SPHH groups; the SPHL, SPHM, and SPHH groups were treated with SPH at concentrations of 50, 100, and 200 mg·L-1, respectively, at the same time. After 24 and 48 h of culture, the reactive oxygen species (ROS) level of the HSF cells was detected by flow cytometry, and the required culture time of the HSF cells for the follow-up experiment was selected. The malondialdehyde and glutathione contents, as well as the activities of the superoxide dismutase, catalase, and glutathione peroxidase in the HSF cells, were detected by biochemical methods. The levels of expression of MMP-1 and collagen I protein in HSF cells were detected by the western blot test, the extent of aging of HSF cells was detected by ß-galactosidase staining, and the apoptosis level of HSF cells was detected by flow cytometry. The results show that SPH inhibits the UVA-induced photoaging of HSF cells in a dose-dependent manner within a certain concentration range, and the effect of a concentration of 200 mg·L-1 was the most significant. The mechanism is related to improving the antioxidant activity of photoaging HSF cells to eliminate excessive ROS. It can inhibit apoptosis, reduce the protein expression of MMP-1, and effectively control the degradation of collagen I protein in photoaging HSF cells. Therefore, SPH offers potential for use in sunscreen cosmetics.

Effects of different silicate minerals on silicon activation by Ochrobactium sp. T-07-B.

As a kind of solid waste with a high silicon content, electrolytic manganese residue (EMR) can be utilized as silicon source by plants through bioleaching processes. EMR contains a variety of silicate minerals. In order to determine the source of available silicon in the bioleaching process of EMR, it is necessary to investigate the influence of silicate minerals in EMR on silicon-activating behavior of specific minerals. In this study, Ochrobactium sp. T-07-B was used to conduct bioleaching experiments on five kinds of silicate minerals with different structures (quartz, muscovite, biotite, olivine, and rhodonite); the growth of Ochrobactium sp. T-07-B, their acid- and polysaccharide-producing capacity, and evolution of surface morphology and structure of the silicate minerals in different systems were determined, so as to explore the silicon-activating capacity of Ochrobactium sp. T-07-B and the selectivity toward different minerals in the bioleaching process. Results showed that the effects of Ochrobactium sp. T-07-B for different silicate minerals were obviously different, and the sequence of silicon-activating efficiency from high to low was as follows: muscovite (65.84 mg·L-1) > biotite (63.84 mg·L-1) > olivine (55.76 mg·L-1) > rhodonite (50.98 mg·L-1) > quartz (23.63 mg·L-1). Results of this study may be of guiding significance for the future research on the silicon-activating behavior of solid waste.

Fast, Simple, and Highly Specific Molecular Detection of Porphyromonas gingivalis Using Isothermal Amplification and Lateral Flow Strip Methods.

Porphyromonas gingivalis is an important oral pathogen that causes periodontal disease and is difficult to culture under conventional conditions. Therefore, a reliable technique for detecting this pathogenic bacterium is required. Here, isothermal recombinase polymerase amplification (RPA), a new nucleic acid amplification method, was combined with a visualization method based on nanoparticle-based lateral flow strips (LFS) for the rapid detection of P. gingivalis. The species-specific 16S rRNA sequence of P. gingivalis was used as the target for RPA, and a set of specific primer-probe combinations were designed and screened to amplify the target sequences. As a thermostatic amplification method, the RPA reaction, under optimized conditions, takes only 30 min to complete at a constant temperature (37°C). The amplification reaction products can be detected visually by LFS without any need for special equipment. The RPA-LFS method established for the detection of P. gingivalis was shown to be highly specific in distinguishing P. gingivalis from other pathogenic organisms by using 20 clinical isolates of P. gingivalis and 23 common pathogenic microorganisms. Susceptibility measurements and probit regression analysis were performed with gradient dilutions of P. gingivalis genomic DNA. The method was obtained to be highly sensitive, with a detection limit of 9.27 CFU per reaction at 95% probability. By analyzing the gingival sulcus fluid specimens from 130 patients with chronic periodontitis, the results showed that the RPA-LFS method detected 118 positive cases and 12 negative cases of P. gingivalis, and the results obtained were consistent with those of a conventional PCR assay. The RPA-LFS method is an efficient, rapid, and convenient diagnostic method that simplifies the tedious process of detecting P. gingivalis.

Protective Effect of the Pearl Extract from Pinctada fucata martensii Dunker on UV-Induced Photoaging in Mice.

Although the effect of pearl powder has been recognized for more than a thousand years from healthcare to beauty care, there has yet to be an in-depth understanding of its anti-photoaging effect. In the present study, the protective effect of pearl extract (PE) on UV-induced photoaging in mice was evaluated. First, the amino acid analysis of PE was carried out. Then, different dosages of pearl extract gel (PEG) were applied topically on the shaved dorsal skins regions of mice before UV irradiation. Skin physiological and histological analysis, antioxidant enzymes and inflammatory factor test were used to evaluate the anti-photoaging effect of PEG. The results showed that PEG contained 14 amino acids, and could inhibit UV-irritated skin wrinkles, laxity, thickness, and dryness. Moreover, PEG upregulated the activities of CAT, GSH-Px, SOD and decreased MDA level, and suppressed the production of IL-1ß, IL-6, PGE2 , TNF-α, and COX-2 in UV-irradiated mice. The therapeutic effect in high dose PEG group was superior to those of positive control (Vitamin E). This study demonstrated the underlying mechanisms of PEG against UV-irritated photoaging. And PEG possesses a potential use in photoprotective medicines and cosmetics.

Therapeutic Effect of Seawater Pearl Powder on UV-Induced Photoaging in Mouse Skin.

The objective of this study was to investigate the therapeutic effect of seawater pearl powder (SPP) on ultraviolet (UV) irradiation-induced photoaging in mouse skin. The protein and trace elements in SPP were detected by liquid chromatography-mass spectrometry, atomic fluorescence spectrometry, and inductively coupled plasma-atomic emission spectrometry. The effect of SPP on treating skin damage resulting from UV-induced photoaging was observed by gross physical appearance and histopathological analysis. Oxidative stress and melanin synthesis were analyzed using biochemical method. Western blotting was applied to analyze the phosphorylation and expression levels of matrix metalloproteinase-1 (MMP-1), collagen I, and proteins involved in the mitogen-activated protein kinase (MAPK) signaling pathways (p38, ERK, and JNK). The results show that SPP has a significant therapeutic effect on UV-induced photoaging of skin and improves and restores appearance and tissue structure of mouse skin. The major mechanism may be related to reduction of expression level of MMP-1 and enhancement of collagen I production via inhibition of MAPK signaling pathway after scavenging of excess reactive oxygen species (ROS) in the UV-induced photoaged skin of mice. Meanwhile, it may also be involved in reducing melanin content by inhibiting tyrosinase activity after scavenging excess ROS in the UV-induced photoaged skin of mice. Therefore, SPP could be a good substance to treat photoaging skin. Taking cost-effectiveness and efficacy into consideration, the optimal concentration of SPP for treating photoaging skin could be 100 mg/g.

Development and validation of a nomogram to predict postoperative pulmonary complications following thoracoscopic surgery.

BACKGROUND: Postoperative pulmonary complications (PPCs) after thoracoscopic surgery are common. This retrospective study aimed to develop a nomogram to predict PPCs in thoracoscopic surgery. METHODS: A total of 905 patients who underwent thoracoscopy were randomly enrolled and divided into a training cohort and a validation cohort at 80%:20%. The training cohort was used to develop a nomogram model, and the validation cohort was used to validate the model. Univariate and multivariable logistic regression were applied to screen risk factors for PPCs, and the nomogram was incorporated in the training cohort. The discriminative ability and calibration of the nomogram for predicting PPCs were assessed using C-indices and calibration plots. RESULTS: Among the patients, 207 (22.87%) presented PPCs, including 166 cases in the training cohort and 41 cases in the validation cohort. Using backward stepwise selection of clinically important variables with the Akaike information criterion (AIC) in the training cohort, the following seven variables were incorporated for predicting PPCs: American Society of Anesthesiologists (ASA) grade III/IV, operation time longer than 180 min, one-lung ventilation time longer than 60 min, and history of stroke, heart disease, chronic obstructive pulmonary disease (COPD) and smoking. With incorporation of these factors, the nomogram achieved good C-indices of 0.894 (95% confidence interval (CI) [0.866-0.921]) and 0.868 (95% CI [0.811-0.925]) in the training and validation cohorts, respectively, with well-fitted calibration curves. CONCLUSION: The nomogram offers good predictive performance for PPCs after thoracoscopic surgery. This model may help distinguish the risk of PPCs and make reasonable treatment choices.

Embelia Laeta aqueous extract suppresses acute inflammation via decreasing COX-2/iNOS expression and inhibiting NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Embelia laeta (L.) Mez., which is called Suanjifeng in Chinese ethnic Yao medicine, is traditionally for inflammation-related diseases, such as oral ulcer, sore throat, enteritis, and rheumatoid arthritis. However, the biological properties and the underlying mechanisms of Embelia laeta still need further studies. AIM OF THIS STUDY: The present study aims to investigate the anti-inflammatory effect and its underlying mechanisms of Embelia laeta. MATERIALS AND METHODS: In this study, except acute toxicity experiments, Kunming (KM) mice of either sex were enrolled to establish inflammatory model induced by xylene, acetic acid and carrageenan, respectively. Mice were randomly divided into different groups and pretreated by oral gavage with different doses of Embelia laeta aqueous extract (ELAE) (2.5, 5, 10 g/kg) and 10 mg/kg of Indo for 7 days. Ear edema, vascular permeability, abdominal writhing, and paw edema degree were detected in related experiments. Moreover, in the carrageenan-induced paw edema mice model, histological changes were detected by H&E staining. MDA, MPO and NO were detected by assay kit. Proinflammatory cytokines of IFN-γ, TNF-α, IL-1ß, IL-6 and PGE2 were detected by ELISA. Additionally, COX-2, iNOS and NF-κB pathway-related proteins were detected by Western blotting. RESULTS: Results showed that the ELAE evoked an obvious dose-dependent inhibition of ear edema induced by xylene, paw edema induced by carrageenan, as well as suppressing the increase of vascular permeability and writhing times elicited by acetic acid. Histopathological analysis indicated that ELAE could significantly decrease the cellular infiltration in paw tissue. ELAE showed antioxidant property through markedly decrease the MDA level and MPO activity in edema paw. In addition, ELAE decreased the proinflammatory cytokines IFN-γ, TNF-α, IL-1ß, IL-6, PGE2 and NO that induced by carrageenan. Western blotting results also showed that ELAE could obviously downregulate the COX-2 and iNOS expression. Further analysis revealed that ELAE also inhibited NF-κB from the cytoplasm to the nucleus and stabilize the conversion of IκBα. CONCLUSION: ELAE had powerful anti-inflammatory property, which might be had a close relationship with mediating proinflammatory cytokines production, decreasing the COX-2 and iNOS expression, and inhibiting the activation of NF-κB signaling pathway.

Screening of silicon-activating bacteria and the activation mechanism of silicon in electrolytic manganese residue.

Electrolytic manganese residue (EMR) is a kind of solid waste with a high silicon content. Most of the silicon in EMR, however, exist in the state of SiO2, which cannot be directly absorbed by plants. Currently, it is very challenge to recover the silicon from EMR. In this study, a preliminary screening of strains with silicon-activating ability was conducted, and four strains were screened out and isolated from the soil around the tailings pond of EMR. Then, single factor experiments were conducted to obtain the optimal growth conditions of the four strains, and the results indicated that the Ochrobactrum sp. T-07 had the best silicon-activating ability from EMR after nitrosoguanidine mutagenesis (Ochrobactrum sp. T-07-B). The available silicon (in terms of SiO2) in the leaching solution was up to 123.88 mg L-1, which was significantly higher than that produced by Bacillus circulans and Paenibacillus mucilaginosus, the two commercial available pure culture strains. Results of direct/indirect contact experiments between Ochrobactrum sp. T-07-B and EMR revealed that bioleaching was promoted under the synergistic effect of bacteria growth on the surface of and metabolism within EMR. The newly isolated strains with silicon-activating effect are different from the existing-known silicate bacteria and may be used for more efficient silicon activation in silicate minerals.

Hydrolyzed seawater pearl tablet modulates the immunity via attenuating Th1/Th2 imbalance in an immunosuppressed mouse model.

OBJECTIVE: To investigate whether Hydrolyzed Seawater Pearl tablet (HSPT) could modulate the Th1/Th2 imbalance in an immunosuppressed mouse model with Th1 to Th2 shift induced by Cyclosporine A (CsA) which can be used in the clinical treatment of Th2 to Th1 shift diseases, and explore the possible mechanism for the adjuvant therapeutic efficacy of HSPT on recurrent respiratory infections (RRI) and acquired immune deficiency syndrome (AIDS). METHODS: The mice were randomly divided into six groups of five animals each, namely normal group, model group, lentinan polysaccharide tablet (LPT) group and three HPST treated groups. HPST treated groups were administered with HPST (0.51, 1.02, 2.04 g/kg) via intragastric gavage (i.g) for 30 consecutive days. LPT used as reference drug for positive control, LPT group was administered with LPT (8.2 mg/kg) for 30 consecutive days. Normal group and model group were received distilled water. The animals in model group, LPT group and HPST treated groups were injected intraperitoneally with CsA (50 mg/kg) to establish the immunosuppressed mice model with Th1 to Th2 shift on the 20th, 22nd and 24th day, one hour after the administration of the respective treatment. Animals were sacrificed one hour after the last administration to collect blood and splenic tissue. The proportion of T cells including CD8+ and CD4+ T cells, Th1 and Th2 in peripheral blood of experimental mice were measured by flow cytometric. The protein level in serum and mRNA level in splenic tissue of experimental mice for interleukin (IL)-2, IL-12, interferon-γ (IFN-γ), IL-4, IL-6, IL-10 and IL-13 were measured by enzyme linked immunosorbent assay and fluorescence quantitative polymerase chain reaction respectively. RESULTS: HSPT elevated the proportion of T cells including both CD8+ and CD4+ T cells, in which the proportion of Th1 and Th2 cells increased, while the ratio of Th1/Th2 cells decreased in peripheral blood of the immunosuppressed mouse model with Th1 to Th2 shift induced by CsA. Furthermore, HSPT elevated both protein and mRNA level of Th1-type cytokines IL-2 and IFN-γ, while had no significant effect on protein and mRNA level of Th1-type cytokine IL-12 and Th2-type cytokines IL-4, IL-6, IL-10, IL- 13 in mouse model. CONCLUSION: Our findings suggest that HSPT can increase proportion of T cells including both CD8+ and CD4+ T cells and induce Th2 to Th1 shift in both cells and cytokines, which probably was the mechanism to account for the adjuvant therapeutic efficacy of HSPT on RRI and AIDS.

An optimized method for the induction and purification of mouse bone marrow dendritic cells.

Dendritic cells (DCs) play an essential role in the initiation of adaptive immune responses, but they are rare in all organs. The traditional methods used to increase the yield and purity of DCs are the early removal of granulocyte culture medium and the isolation of high-purity DCs by magnetic-activated cell sorting (MACS). This study provides a more rapid and economical optimization method to obtain more high-purity DCs. (i) We harvested 18% more bone marrow (BM) cells by using forceps to crack the epiphysis instead of cutting it with scissors during BM cell extraction. (ii) When the cells in the culture medium that is discarded on day 3 in the traditional method were centrifuged and then added back to the petri dish, the DC yield on day 5 increased by 61%. (iii) On the third day, the addition of fresh medium and the retention of the original medium rather than discarding it increased the number of DCs harvested on the fifth day by 137%. (i-iii) The improved method cost an average of 74% less than the conventional method and yielded the same number and function of cells. (iv) The initial number of BM cells was increased by 15% in 4-week-old mice compared with 8-week-old mice. (v) The Percoll density centrifugation (PDS) method was used to purify DCs on day 6 after induction, and the purity of the DCs was greater than 90%, which showed no significant difference from the MACS method. However, the yield of the PDS method increased by 21%. In addition, the PDS method has a lower cost, with an average purification cost of 4 CNY ($0.58) compared with 648 CNY ($93.25) for MACS, reducing the cost by 99%. Therefore, high-purity and high-yield DCs can be rapidly obtained through a five-step improvement in the process of BM cell extraction, induction and purification.

(+/-)-Borneol Reverses Mitoxantrone Resistance against P-Glycoprotein.

P-Glycoprotein (Pgp) is a main factor contributing to multidrug resistance and the consequent failure of chemotherapy. Overcoming Pgp efflux is a strategy to improve the efficacy of drugs. (+)-Borneol (BNL1) and (-)-borneol (BNL2) interfere and inhibit Pgp, and thus, the accumulation of drugs increases in cells. However, it is not clear yet how they play the inhibitory effect against Pgp. In this work, the effect and molecular mechanism of borneol enantiomers in reversing mitoxantrone (MTO) resistance against Pgp were explored by in vitro and in silico approaches. Chemosensitizing potential tests showed that BNLs could enhance the efficacy of MTO in MES-SA/MX2 cells, and BNL2 exhibited a stronger potential. The protein expression of Pgp was decreased to some extent by the administration of BNLs. Molecular docking revealed that BNLs could reduce the binding affinity between MTO and Pgp. The results were consistent with the chemosensitizing potential test and were supported by molecular dynamics (MD) simulations. Molecular docking also suggested that BNLs preferred to bind in the drug-binding pocket rather than the nucleotide-binding domain of inward-facing Pgp. The occupied space of BNLs had an evident distance from that of MTO, which was further verified by the conformational analysis after MD simulations. The decomposition of binding free energies revealed the key amino acid residues (GLN195, SER196, TRP232, PHE343, SER344, GLY346, and GLN347) for BNLs to reverse MTO resistance. The results provide an insight into the mechanism through which BNLs reduce the MTO resistance against inward-facing Pgp in the drug-binding pocket through noncompetitive inhibition.

Species identification and mutation breeding of silicon-activating bacteria isolated from electrolytic manganese residue.

A strain of silicon-activating bacteria was isolated from electrolytic manganese residue (EMR); identified as a species of Ochrobactrum by integrated microscopic morphological characteristics, biochemical index determination, and clone analysis (i.e., results of 16S rRNA sequence); and temporarily named as Ochrobactrum sp. T-07 (T-07). The optimal growth conditions of the strain T-07 were obtained as follows: temperature of 30 °C, initial pH of 7.0, shaking speed of 180 rev. min-1, and loading volume of 100 mL. In order to enhance its activation activity of silicon, T-07 went through the ultraviolet (UV) mutagenesis and nitrosoguanidine (NTG) mutagenesis breeding, and the mutant strain T-07-B with higher activity was obtained. Under the optimal fermentation condition (leaching time of 20 days, temperature of 30 °C, initial pH of 7, pulp concentration of 5%, shaking speed of 180 rev. min-1, and particle diameter of EMR ≤ 180 µm), the available silicon content in the supernatant reached 98.8 mg L-1, which was 2.4 times of the original strain T-07. Therefore, T-07 can be used as a good backup in developing biological silicon fertilizer for plants.