Phosphorylated vimentin-triggered fibronectin matrix disaggregation enhances the dissemination of Treponema pallidum subsp. pallidum across the microvascular endothelial barrier.

Fibronectin (FN) is an essential component of the extracellular matrix (ECM) that protects the integrity of the microvascular endothelial barrier (MEB). However, Treponema pallidum subsp. pallidum (Tp) breaches this barrier through elusive mechanisms and rapidly disseminates throughout the host. We aimed to understand the impact of Tp on the surrounding FN matrix of MEB and the underlying mechanisms of this effect. In this study, immunofluorescence assays (IF) were conducted to assess the integrity of the FN matrix surrounding human microvascular endothelial cell-1 (HMEC-1) with/without Tp co-culture, revealing that only live Tp exhibited the capability to mediate FN matrix disaggregation in HMEC-1. Western blotting and IF were employed to determine the protein levels associated with the FN matrix during Tp infection, which showed the unaltered protein levels of total FN and its receptor integrin α5ß1, along with reduced insoluble FN and increased soluble FN. Simultaneously, the integrin α5ß1-binding protein-intracellular vimentin maintained a stable total protein level while exhibiting an increase in the soluble form, specifically mediated by the phosphorylation of its 39th residue (pSer39-vimentin). Besides, this process of vimentin phosphorylation, which could be hindered by a serine-to-alanine mutation or inhibition of phosphorylated-AKT1 (pAKT1), promoted intracellular vimentin rearrangement and FN matrix disaggregation. Moreover, within the introduction of additional cellular FN rather than other Tp-adhered ECM protein, in vitro endothelial barrier traversal experiment and in vivo syphilitic infectivity test demonstrated that viable Tp was effectively prevented from penetrating the in vitro MEB or disseminating in Tp-challenged rabbits. This investigation revealed the active pAKT1/pSer39-vimentin signal triggered by live Tp to expedite the disaggregation of the FN matrix and highlighted the importance of FN matrix stability in syphilis, thereby providing a novel perspective on ECM disruption mechanisms that facilitate Tp dissemination across the MEB.

Uncovering the mechanisms of host mitochondrial cardiolipin release in syphilis: Insights from human microvascular endothelial cells.

The release of host mitochondrial cardiolipin is believed to be the main factor that contributes to the production of anti-cardiolipin antibodies in syphilis. However, the precise mechanism by which mitochondria release cardiolipin in this context remains elusive. This study aimed to elucidate the mechanisms underlying mitochondrial cardiolipin release in syphilis. We conducted a cardiolipin quantitative assay and immunofluorescence analysis to detect mitochondrial cardiolipin release in human microvascular endothelial cells (HMEC-1), with and without Treponema pallidum (Tp) infection. Furthermore, we explored apoptosis, a key mechanism for mitochondrial cardiolipin release. The potential mediator molecules were then analyzed through RNA-sequence and subsequently validated using in vitro knockout techniques mediated by CRISPR-Cas9 and pathway-specific inhibitors. Our findings confirm that live-Tp is capable of initiating the release of mitochondrial cardiolipin, whereas inactivated-Tp does not exhibit this capability. Additionally, apoptosis detection further supports the notion that the release of mitochondrial cardiolipin occurs independently of apoptosis. The RNA-sequencing results indicated that microtubule-associated protein2 (MAP2), an axonogenesis and dendrite development gene, was up-regulated in HMEC-1 treated with Tp, which was further confirmed in syphilitic lesions by immunofluorescence. Notably, genetic knockout of MAP2 inhibited Tp-induced mitochondrial cardiolipin release in HMEC-1. Mechanically, Tp-infection regulated MAP2 expression via the MEK-ERK-HES1 pathway, and MEK/ERK phosphorylation inhibitors effectively block Tp-induced mitochondrial cardiolipin release. This study demonstrated that the infection of live-Tp enhanced the expression of MAP2 via the MEK-ERK-HES1 pathway, thereby contributing to our understanding of the role of anti-cardiolipin antibodies in the diagnosis of syphilis.

[Analysis of Bacterial Communities and Antibiotic Resistance Genes in the Aquaculture Area of Changli County].

In order to understand the distribution of microorganisms and various antibiotic resistance genes in the aquaculture area of Changli County, Qinhuangdao, high-throughput sequencing technology was used in this study. We utilized 16S rDNA gene sequencing and metagenome sequencing methods to analyze the seawater, sediment, and gut contents of the local fish Synechogobius hasta in the aquaculture area in spring. The results showed that Proteobacteria, Firmicutes, and Bacteroidota were the dominant bacteria in seawater; and Proteobacteria, Crenarchaeota, Acidobacter, and Actinobaciota were rich in the sediment; whereas Proteobacteria, Cyanobacteria, Firmicutes, and Bacteroidota were in relatively high abundance in fish gut contents. The microbial diversity of sediment samples was the most abundant, followed by seawater samples, and the microbial diversity of fish intestinal contents was the lowest. Moreover, the microbial diversity of similar samples was relatively similar, and the microbial diversity of different types of samples was quite different. For samples at different sites, there were significant differences between seawater samples at each site, and there were small differences between sediment samples at each site, and some sediment sample groups did not have significant differences in microbial composition. In all sample groups, five ß-lactam antibiotic resistance genes (blaOXA-325, cepS, blaCARB-20, blaOXA-55, and blaTRU-1) and four aminoglycoside antibiotic resistance genes[aac(6')-IIb, amrA, aac(6')-Ie-aph(2″)-Ia, and aph(3')-Vc] were detected. There was also a certain correlation between antibiotic resistance genes and microbial communities.

The m6 A modification of Il17a in CD4+ T cells promotes inflammation in psoriasis.

Psoriasis is a chronic inflammatory skin disorder. The mechanism of psoriasis pathogenesis is not entirely clear. Here, we reported that the level of the N6-methyladenosine (m6 A) modification was increased in psoriatic CD4+ T cells compared with healthy controls. In the psoriasis mouse model, depletion of the RNA demethylase, Alkbh5, from CD4+ T cells promoted the psoriasis-like phenotype and inflammation. Intriguingly, this phenotype and inflammation were alleviated by the ablation of the m6 A methyltransferase Mettl3 in CD4+ T cells. Mechanistically, we found that the m6 A modification of IL17A mRNA increased the expression of IL-17A (an important pro-inflammatory factor in psoriasis) and promoted psoriasis. Thus, our study provided evidence that the m6 A modification of IL17A in CD4+ T cells regulates inflammation in psoriasis.

Recent advances in targeting leucine-rich repeat kinase 2 as a potential strategy for the treatment of Parkinson's disease.

Parkinson's disease (PD) is the second most common neurodegenerative disease. Several single gene mutations involved in PD have been identified such as leucine-rich repeat kinase 2 (LRRK2), the most common cause of sporadic and familial PD. Its mutations have attracted much attention to therapeutically targeting this kinase. To date, many compounds including small chemical molecules with diverse scaffolds and RNA agents have been developed with significant amelioration in preclinical PD models. Currently, five candidates, DNL201, DNL151, WXWH0226, NEU-723 and BIIB094, have advanced to clinical trials for PD treatment. In this review, we describe the structure, pathogenic mutations and the mechanism of LRRK2, and summarize the development of LRRK2 inhibitors in preclinical and clinical studies, trying to provide an insight into targeting LRRK2 for PD intervention in future.

Mixed oligosaccharides-induced changes in bacterial assembly during cucumber (Cucumis sativus L.) growth.

The application of oligosaccharides can promote plant growth by increasing photosynthesis or inducing plant innate immunity. However, the mechanisms by which oligosaccharides affect bacterial community diversity and abundance remain unclear. In this study, a mixed oligosaccharide was applied to the growth of cucumbers. The findings of the present study suggest that the application of MixOS has significant effects on the bacterial communities in the phyllosphere, rhizosphere, and bulk soil of cucumber plants. The treatment with MixOS resulted in delayed senescence of leaves, well-developed roots, and higher fruit production. The bacterial diversity and composition varied among the different ecological niches, and MixOS application caused significant shifts in the bacterial microbiome composition, particularly in the phyllosphere. Moreover, mixed oligosaccharides increased the abundance of potential growth-promoting bacteria such as Methylorubrum spp. and Lechevalieria spp., and more zOTUs were shared between the WM and MixOS treatments. Furthermore, the bacterial co-occurrence network analysis suggested that the modularity of the phyllosphere networks was the highest among all samples. The bacterial co-occurrence networks were altered because of the application of MixOS, indicating a greater complexity of the bacterial interactions in the rhizosphere and bulk soil. These findings suggest that mixed oligosaccharides has the potential to improve plant growth and yield by modulating the bacterial communities within and outside the plants and could provide a theoretical basis for future agricultural production.

Psoriasis and COVID-19 Infection Negatively Impact Each Other: An Analysis of 3581 Cases.

Objective: To determine whether COVID-19 infection and psoriasis impact each other. Methods: WeChat app was used to carry out a questionnaire survey in individuals aged 18 years and over. Results: A total of 3581 individuals, including 2515 individuals without psoriasis and 1066 psoriatic patients completed the survey. The rate of COVID-19 infection was significantly higher in psoriatic patients than in those without psoriasis (89.59% vs 77.93%, p < 0.0001). Moreover, the rate of relapse and/or worsening of psoriasis was higher in psoriatic patients with COVID-19 infection than in that without COVID-19 infection (75.29% vs 47.75%, p < 0.0001). Conclusion: COVID-19 infection and psoriasis negatively impact each other.

Effect of Individualized Cardiac Rehabilitation on Cardiac Function, Time Consumption, and Quality of Life in Patients After CABG.

BACKGROUND: To investigate the effect of individualized cardiac rehabilitation (CR) on cardiac function, time consumption, and quality of life (QoL) in post-CABG patients. METHODS: Two different CR strategy: basic rehabilitation and individualized rehabilitation was designed. The patients were screened and randomized into the two groups: the basic rehabilitation group (BRG) and individualized rehabilitation group (IRG). Data, such as clinical characteristics, LVEF, 6MWD (6-min walk distance), BNP, LVEDD (left ventricular end diastolic dimension), SF-36 score, and time consumption were collected and recorded. RESULTS: There was no difference between the IRG and BRG patients in the clinical characteristics. The 6MWD and LVEF on post-op significantly were higher, while BNP and LVEDD significantly was lower in the IRG than in BRG. The time to first out-of-bed activity, ICU stay time, and post-op hospital stay time of the IRG in post-op was significantly shorter than BRG. The IRG patients scored significantly higher on the SF-36. CONCLUSION: Individualized CR is safe and can reduce the time consumption and improve the cardiac function and QoL of patients undergoing CABG.

Generation of a human embryonic stem cell line (SMUDHe010-A-82) carrying a homozygous c.1538G > A (p.G513D) mutation in the OSMR gene by CRISPR/Cas9-mediated homologous recombination.

Mutations in the tumor suppressor M receptor (OSMR) gene are associated with primary localized cutaneous amyloidosis (PLCA). Recently, we confirmed that OSMR loss-of-function mutations enhance epidermal keratinocyte differentiation via inactivation of the STAT5/KLF7 signaling. However, no disease model was available for PLCA. Accordingly, we generated an OSMR c.1538G > A mutant human embryonic stem cell line (SMUDHe010-A-82) using CRISPR/Cas9-mediated homologous recombination. The cell line preserves normal karyotype, pluripotency and the ability to differentiate into all three germ layers. Moreover, the cell line can be used to prepare human skin organoid, which may provide a disease model for PLCA.

Diagnosis of Primary Cutaneous Amyloidosis by Rapid 4,6-Diamidino-2-Phenylindole Staining.

BACKGROUND: Quick and accurate diagnosis of primary cutaneous amyloidosis (PCA) may be difficult because its symptoms are often subtle and nonspecific. OBJECTIVE: We sought to review the literature on the roles of various staining methods in the diagnosis of amyloidosis and demonstrate added benefits of using rapid 4,6-diamidino-2-phenylindole (DAPI) staining in the diagnosis of PCA. METHODS: Three groups of cases, namely, PCA, neurodermatitis, and prurigo nodularis, were retrieved from a computerized pathology database for study, and their paraffin-embedded tissue blocks were cut following standard procedures. The tissue sections were stained with three stains: hematoxylin-eosin (HE), Congo red, and DAPI stains, and examined under the microscope to compare the staining patterns of these three methods. We also performed amyloid keratin and apolipoprotein E (APOE) staining on the sections of PCA in order to further support our conclusion. The PCA sections were read by junior and senior dermatopathologists for comparison. RESULTS: The sensitivity of DAPI staining for PCA was significantly higher than that of Congo red staining and HE staining (p < 0.001). This statement holds true whether the experiment was grouped in one sample or was divided into groups of junior and senior dermatopathologists (p < 0.001). The DAPI-positive staining areas, except for the nuclei, were consistent with the amyloid deposition areas. In this study, DAPI staining had a sensitivity of 98.6% and a specificity of 100%. CONCLUSION: DAPI staining could serve as a useful technique to establish the diagnosis of PCA, and its high efficacy in diagnosing PCA makes it less dependent on the experience levels of the evaluators. Additionally, the binding of DAPI to the A-T-rich sequence of double-stranded DNA suggests that amyloid may contain DNA or a similarly structured nucleic acid.

Ameliorated membranous nephropathy activities of two ethanol extracts from corn silk and identification of flavonoid active compounds by LC-MS2.

The current study looks to evaluate the effect of corn silk flavonoids on membranous nephropathy (MN). Polyamide resin (PR) can be used to enrich corn silk ethanol extract (CSEE) to obtain flavonoid-rich extract (PR-CSEE), the total flavonoid content (TFC) of which we found to be 57.4%. The results of scanning electron microscope, Fourier-transform infrared, and high-performance liquid chromatography analyses determined that PR-CSEE and CSEE have different structural characteristics, but that PR-CSEE has higher TFC. MN mice models were induced by cationic bovine serum albumin, and we found that PR-CSEE administration reduced urine protein levels markedly, while renal function, glomerular atrophy, inflammatory infiltration, and in-serum immunoglobulin G and complement 3 content were improved. Through LC-MS2 spectrometry analysis, we pinpointed the 12 major flavonoid active compounds in PR-CSEE. These findings suggest that PR-CSEE can act as a potential functional food material by which to improve MN.

A retrospective study to assess the efficacy, safety, and drug survival of secukinumab in plaque psoriasis patients in China.

Secukinumab, a full human immunoglobulin G1κ monoclonal antibody that targets interleukin-17A, has demonstrated remarkable efficacy and appreciable tolerance in patients with moderate-to-severe psoriasis. However, data on its real-life performance, particularly on drug survival in China are limited. To investigate the efficacy, safety, and drug survival of secukinumab in Chinese patients with psoriasis, we conducted a monocentric retrospective study of 66 patients with moderate-to-severe psoriasis to followed-up for 52 weeks. At week 12, 86.4%, 57.6%, and 10.6% of the patients attained 75% improvement in psoriasis area and severity Index (PASI) score from baseline (PASI 75), PASI 90, and PASI 100 responses, respectively. The quality of life of patients markedly improved. The overall survival rate was 74.2%. Adverse events occurred in 30 patients (45.5%). The results revealed favorable efficacy, safety, and tolerability of secukinumab in the treatment of patients with psoriasis and provided data on drug survival in real-life clinical setting in China for the first time.

Hypouricemia effects of corn silk flavonoids in a mouse model of potassium oxonated-induced hyperuricemia.

The hypouricemic effect of corn silk flavonoids (CSFs) in vivo that were extracted by ethanol and fractionated by continuous elution with 40% (CSF-A) and 60% (CSF-B) ethanol solutions on polyamide column were investigated in this study. CSFs reduced serum uric acid (UA) level in a mouse model of potassium oxonate-induced hyperuricemia. CSF-B had the best hypouricemic effect, as it decreased the serum UA level by 26.69% and xanthine oxidase (XO) activity in the serum by 11.29%. The mechanism of action of CSF-B was related to the inhibition of XO activity and the promotion of UA excretion. CSF-B was found to contain 12 kinds of major flavonoids, five of which were speculated to influence its activity in the hyperuricemia mice. The five flavonoids were apigenin-6-C-glucoside-7-O-glucoside, kaempferol-3-O-rutinoside, luteolin-7-glucoside, luteolin-3',7-di-O-glucoside, and naringenin, respectively. Structure analysis revealed that C-4', C5 hydroxyl groups, and C2=C3 double bonds in CSF-B gave the latter its hypouricemic effect. PRACTICAL APPLICATIONS: The prevalence of hyperuricemia has increased in recent times. Current hypouricemic drugs have side effects and can easily lead to various complications. Therefore, it is of great practical significance to find safer and more effective hypouricemic drugs. This study demonstrated that corn silk flavonoids may be used as a dietary supplement to manage hyperuricemia.

Exploration on self-equilibrium rule and adsorption-desorption model between pine nut (Pinus koraiensis) peptide molecules and environmental moisture molecules.

The storage of pine nut (Pinus koraiensis) peptide (PNP) powder involves hygroscopic phenomena. To investigate the adsorption and self-equilibrium rules between these peptides and the environmental moisture molecules, several studies such as low-field nuclear magnetic resonance (LF-NMR), dynamic vapor sorption (DVS) and adsorption-desorption models were done. The results showed that the outward moisture migration occurred during storage as 7.80% and 16.68% moisture were respectively constrained by the original sample and 90 days after lyophilization, by chemical bonding. Additionally, 1.79% moisture was lost in PNP powder at day 90. The optimized adsorption model for PNP powder was changed from Henderson's to Oswin's model during the 90 days' storage whereas the optimized desorption model was changed from Halsey's to GAB's model. The PNP powder at day 90 presented smaller particles with an average diameter and height of 15.645 nm and 50 nm, respectively, and it contained more molecular moisture which cannot be removed. The free thiol of the PNP powder at day 0 and day 90 was 1.75 ± 0.16 µM SH/g and 1.95 ± 0.16 µM SH/g, respectively, and the total sulfhydryl was 101.46 ± 1.06 µM SH/g and 118.44 ± 1.27 µM SH/g. The registered increased sulfhydryl content contributed to the generation of off-flavor.

Antifungal curcumin promotes chitin accumulation associated with decreased virulence of Sporothrix schenckii.

Curcumin, a yellow polyphenol compound, is known to possess antifungal activity for a range of pathogenic fungi. However, the fungicidal mechanism of curcumin (CUR) has not been identified. We have occasionally found that chitin redistributes to the cell wall outer layer of Sporothrix schenckii (S. schenckii) upon sublethal CUR treatment. Whether CUR can affect chitin synthesis via the protein kinase C (PKC) signaling pathway has not been investigated. This study describes a direct fungicidal activity of CUR against S. schenckii demonstrated by the results of a checkerboard microdilution assay and, for the first time, a synergistic effect of CUR with terbinafine (TRB). Furthermore, the results of real-time PCR showed that sublethal CUR upregulated the transcription of PKC, chitin synthase1 (CHS1), and chitin synthase3 (CHS3) in S. schenckii. The fluorescence staining results using wheat germ agglutinin-fluorescein isothiocyanate (WGA-FITC) and calcofluor white (CFW) consistently showed that chitin exposure and total chitin content were increased on the conidial cell wall of S. schenckii by sublethal CUR treatment. A histopathological analysis of mice infected with CUR-treated conidia showed dampened inflammation in the local lesion and a reduced fungal burden. The ELISA results showed proinflammatory cytokine secretion at an early stage from macrophages stimulated by the CUR-treated conidia. The present data led to the conclusion that CUR is a potential antifungal agent and that its fungicidal mechanism may involve chitin accumulation on the cell wall of S. schenckii, which is associated with decreased virulence in infected mice.

Taenia taeniaeformis in rat favors protracted skin lesions caused by Sporothrix schenckii infection: Dectin-1 and IL-17 are dispensable for clearance of this fungus.

We occasionally found that cestode Taenia taeniaeformis in rats favored Sporothrix schenckii infection and survival, causing protracted cutaneous lesions. In this study, we compared the pathology and cytokines profile of rats co-infected with the two pathogens and infected with S. schenckii alone to explore underlying mechanisms. In the co-infection group, there was high expression of ß-glucan receptor Dectin-1 in the cutaneous lesions and no multinucleated giant cells, but in the S. schenckii infection group the opposite was observed. Cytokines profiles demonstrated an expected finding that IL-4, commonly expressed in helminth and fungus infection, is undetectable in the two infection groups. In the single fungal infection group, cytokines IFN-γ, IL-10 and IL-17 kept increasing in the first few weeks of infection to a peak which was followed by gradual decrease. This study showed that Dectin-1 and IL-17, which were believed to be the major anti-fungus mechanisms, are Th2 independent and dispensable for clearance of S. schenckii infection, suggesting that S. schenckii has a different molecular recognition pattern and evokes anti-infection mechanisms other than Dectin-1 and IL-17.

[Study on sensitization and mechanism of CGA-BSA].

OBJECTIVE: To investigate the sensitization and mechanism of artificial antigen of chlorogenic acid (CGA-BSA). METHOD: Using intensive immunization to establish allergy animal model on guinea pig and preparing antiserum and tissue for further test. Using HE staining to observe pathology change of lungs, trachea, liver. Using passive mast cell (PMC) degranulation test to observe the immunogenicity of CGA-BSA and using ELISA to detect IgE and histamine in plasma. RESULT: There established allergy animal model on guinea pig, which include a increase cell degranulation by a ratio (63.58 +/- 10.23)% in PMC test, increase of specific antibody IgE and increase of histamine in plasma after provocation by ELISA. CONCLUSION: Allergen CGA-BSA could provoke allergenic response in guinea pig, and the allergic response belongs to type I allergy.

[Study of immunogenicity of chlorogenic acid-BSA with different number of CGA].

OBJECTIVE: To investigate the immunogenicity of chlorogenic acid-BSA(CGA-BSA) and the foundation for the allergization of CGA-BSA. METHOD: The CGA-BSA with different number of CGA was synthesized to allergize the BALB/c mice and SD rats to get antiserum. The level of IgE and histamine was analyzed by ELISA, and the passive cutaneous anaphylaxis (PCA) test was carried to analyze the antibody titer. RESULT: When the coupling rate of CGA-BSA is 20, the highest IgE and histamine level in BALB/c mice and the antibody titer in PCA are the highest. CONCLUSION: It has better immunogenicity when the coupling rate of CGA-BSA is between 10 to 20, while the CGA-BSA 20 with the best immunogenicity.