Heterologous expression of the insect SVWC peptide WHIS1 inhibits Candida albicans invasion into A549 and HeLa epithelial cells.

Candida albicans (C. albicans), a microbe commonly isolated from Candida vaginitis patients with vaginal tract infections, transforms from yeast to hyphae and produces many toxins, adhesins, and invasins, as well as C. albicans biofilms resistant to antifungal antibiotic treatment. Effective agents against this pathogen are urgently needed. Antimicrobial peptides (AMPs) have been used to cure inflammation and infectious diseases. In this study, we isolated whole housefly larvae insect SVWC peptide 1 (WHIS1), a novel insect single von Willebrand factor C-domain protein (SVWC) peptide from whole housefly larvae. The expression pattern of WHIS1 showed a response to the stimulation of C. albicans. In contrast to other SVWC members, which function as antiviral peptides, interferon (IFN) analogs or pathogen recognition receptors (PRRs), which are the prokaryotically expressed MdWHIS1 protein, inhibit the growth of C. albicans. Eukaryotic heterologous expression of WHIS1 inhibited C. albicans invasion into A549 and HeLa cells. The heterologous expression of WHIS1 clearly inhibited hyphal formation both extracellularly and intracellularly. Furthermore, the mechanism of WHIS1 has demonstrated that it downregulates all key hyphal formation factors (ALS1, ALS3, ALS5, ECE1, HWP1, HGC1, EFG1, and ZAP1) both extracellularly and intracellularly. These data showed that heterologously expressed WHIS1 inhibits C. albicans invasion into epithelial cells by affecting hyphal formation and adhesion factor-related gene expression. These findings provide new potential drug candidates for treating C. albicans infection.

[Mechanism of Sijunzi Decoction in treatment of Alzheimer's disease based on UPLC-Q-TOF-MS, network pharmacology, and experimental verification].

The study identified the blood-entering components of Sijunzi Decoction after gavage administration in rats by UPLC-Q-TOF-MS/MS, and investigated the mechanism of Sijunzi Decoction in treating Alzheimer's disease by virtue of network pharmacology, molecular docking, and experimental verification. The blood-entering components of Sijunzi Decoction were identified based on the mass spectra and data from literature and databases. The potential targets of the above-mentioned blood-entering components in the treatment of Alzheimer's disease were searched against PharmMapper, OMIM, DisGeNET, GeneCards, and TTD. Next, STRING was employed to establish a protein-protein interaction(PPI) network. DAVID was used to perform the Gene Ontology(GO) annotation and the Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. Cytoscape 3.9.0 was used to carry out visual analysis. AutoDock Vina and PyMOL were used for molecular docking of the blood-entering components with the potential targets. Finally, the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling pathway enriched by the KEGG analysis was selected for validation by animal experiments. The results showed that 17 blood-entering components were detected in the serum samples after administration. Among them, poricoic acid B, liquiritigenin, atractylenolide Ⅱ, atractylenolide Ⅲ, ginsenoside Rb_1, and glycyrrhizic acid were the key components of Sijunzi Decoction in treating Alzheimer's disease. HSP90AA1, PPARA, SRC, AR, and ESR1 were the main targets for Sijunzi Decoction to treat Alzheimer's disease. Molecular docking showed that the components bound well with the targets. Therefore, we hypothesized that the mechanism of Sijunzi Decoction in treating Alzheimer's disease may be associated with the PI3K/Akt, cancer treatment, and mitogen-activated protein kinase(MAPK) signaling pathways. The results of animal experiments showed that Sijunzi Decoction significantly attenuated the neuronal damage in the hippocampal dentate gyrus area, increased the neurons, and raised the ratios of p-Akt/Akt and p-PI3K/PI3K in the hippocampus of mice. In conclusion, Sijunzi Decoction may treat Alzheimer's disease by activating the PI3K/Akt signaling pathway. The findings of this study provide a reference for further studies about the mechanism of action and clinical application of Sijunzi Decoction.

Treponema pallidum (Syphilis) Antigen TpF1 Induces Activation of Macrophages and Accelerates P2X7R-Induced NLRP3-Dependent Release of IL-1ß.

BACKGROUND: Syphilis is a chronic infectious disease caused by Treponema pallidum (Tp) infection, which causes local inflammation in the host. TpF1 is an oligomeric protein expressed by the Tp-infected host that can induce the host immune response. There are few studies regarding the role of TpF1 in macrophage activation and the subsequent release of cytokines. OBJECTIVE: The objective of this study is to elucidate the effects of TpF1 on the pathological process of Syphilis. In addition, we explored how purinergic 2X7 (P2X7R) induced NOD-like receptor family protein 3 (NLRP3) -dependent release of interleukin-1ß (IL-1ß) and the underlying mechanisms. METHODS: We explored the influence of TpF1 on cytokine release by macrophages using qRT-PCR and ELISA. The specific phenotype of activated macrophages was determined by flow cytometry. RESULTS: TpF1 was able to activate macrophages and induce the M1 macrophage phenotype. Moreover, TpF1 activated the NLRP3 inflammasome in macrophages, which was mediated by P2X7R. CONCLUSION: The Tp-induced protein TpF1 is able to induce macrophage activation and P2X7R-induced NLRP3-dependent release of IL-1ß. Our findings provide a theoretical basis for clarifying the clinical symptoms and pathogenesis of syphilis.

[Relationship between fluoride exposure, orthopedic injuries and bone formation markers in patients with coal-burning fluorosis]. / ç‡ƒç ¤æ±¡æŸ“åž‹æ°Ÿä¸­æ¯’æ‚£è€ æ°Ÿæš´éœ²ã€éª¨ç›¸æŸä¼¤ç¨‹åº¦ä¸Žéª¨å½¢æˆæ ‡å¿—ç‰©çš„å ³ç³».

Chronic exposure to fluoride is a public health problem worldwide. We explored the relationship between fluoride exposure, orthopedic injuries and bone formation markers alkaline phosphatase (ALP), bone Gla protein (BGP) in participants with coal-burning fluorosis in Hehua Village (coal-burning fluorosis endemic area) in Zhijin County of Guizhou Province and Zhangguan Village (non-fluoride contaminated area) in Anshun City of Guizhou Province. Environmental samples were collected and fluoride contents were examined using a fluoride ion-selective electrode. Dental fluorosis and skeletal fluorosis of 295 participants from Hehua Village and 85 participants from Zhangguan Village were diagnosed with informed consent. Urinary samples and peripheral blood samples were collected from the participants to determine urinary fluoride (UF), ALP acti-vity, and BGP content. The results showed that fluoride contents in rice, pepper, corn, drinking water, clay, vegetable-grown soil, coal and indoor and outdoor air were significantly higher than those in the control area, but lower than the previously reported values. With the increases of UF concentration, the ALP activity and BGP content significantly increased, the severity of skeletal fluorosis was greater, but with no significant changes in dental fluorosis. There was positive correlation between the severity of skeletal fluorosis and ALP activity, BGP content. These results indicated that low fluoride exposure could cause orthopedic injuries. ALP and BGP could be used to eva-luate the bone turnover in patients with skeletal fluorosis, which would be useful in the auxiliary diagnosis and therapeutic evaluation of skeletal fluorosis.

Quantitative hepatitis B core antibody level is a new predictor for treatment response in HBeAg-positive chronic hepatitis B patients receiving peginterferon.

A recent study revealed that quantitative hepatitis B core antibody (qAnti-HBc) level could serve as a novel marker for predicting treatment response. In the present study, we further investigated the predictive value of qAnti-HBc level in HBeAg-positive patients undergoing PEG-IFN therapy. A total of 140 HBeAg-positive patients who underwent PEG-IFN therapy for 48 weeks and follow-up for 24 weeks were enrolled in this study. Serum samples were taken every 12 weeks post-treatment. The predictive value of the baseline qAnti-HBc level for treatment response was evaluated. Patients were further divided into 2 groups according to the baseline qAnti-HBc level, and the response rate was compared. Additionally, the kinetics of the virological and biochemical parameters were analyzed. Patients who achieved response had a significantly higher baseline qAnti-HBc level (serological response [SR], 4.52±0.36 vs. 4.19±0.58, p=0.001; virological response [VR], 4.53±0.35 vs. 4.22±0.57, p=0.005; combined response [CR], 4.50±0.36 vs. 4.22±0.58, p=0.009)). Baseline qAnti-HBc was the only parameter that was independently correlated with SR (p=0.008), VR (p=0.010) and CR(p=0.019). Patients with baseline qAnti-HBc levels ≥30,000 IU/mL had significantly higher response rates, more HBV DNA suppression, and better hepatitis control in PEG-IFN treatment. In conclusion, qAnti-HBc level may be a novel biomarker for predicting treatment response in HBeAg-positive patients receiving PEG-IFN therapy.

Antithrombotic activities of aqueous extract from Gardenia jasminoides and its main constituent.

CONTEXT: Gardenia jasminoides J. Ellis (Rubiaceae) is a shrub tree species distributed all over the world. Now its pharmacological activities such as anti-atherosclerosis have been extensively studied. OBJECTIVE: To offer pharmacological proof for its further clinical application in cardiovascular diseases, the antithrombotic activities of the aqueous extract of G. jasminoides (GJ-ext) were studied in mouse and rat models. MATERIALS AND METHODS: GJ-ext was administrated orally to detect the effects on the models of carrageenan-induced tail thrombosis and arteriovenous shunt thrombosis. The effects of GJ-ext and geniposide (p.o.) on antiplatelet aggregation were examined. Geniposide and genipin were studied on venous thrombosis by oral administration. RESULTS: GJ-ext (67, 133 and 266 mg/kg) and aspirin (50 mg/kg), respectively, decreased the length of tail thrombus with average thrombus inhibition rate of 21.9, 55.7, 65.8 and 57.6% at 48 h and 19.0, 54.5, 69.3 and 56.9% at 72 h after carrageenan injection and, meanwhile, improved thrombosis induced by arteriovenous shunt (silk thread) with 36.3, 45.5, 86.4 and 63.7% inhibition rate of thrombus respectively, and the ED(50) of GJ-ext was 160.8 mg/kg. Furthermore, GJ-ext (67 mg/kg) and geniposide (20 mg/kg) significantly inhibited platelet aggregation induced by thrombin/collagen with 45.1%/19.3% and 52.8%/26.2% aggregation rate. Geniposide (10-40 mg/kg) and genipin (5-20 mg/kg) inhibited venous thrombosis induced by tight ligation of the inferior vena cava, their ED(50) values were 18.4 and 8.6 mg/kg, respectively. DISCUSSION AND CONCLUSION: This study indicated that GJ-ext and geniposide demonstrated remarkable antithrombotic activities and supported their therapeutic uses for thrombotic diseases.

[Association between IgG antibody against the C-terminal region of the preS1 protein of hepatitis B virus and the early response to interferon therapy in chronic hepatitis B].

OBJECTIVE: To determine the relationship between IgG antibody against the C-terminal region of the preS1 protein of hepatitis B virus and the early response to interferon therapy in chronic hepatitis B. METHODS: 69 patients with chronic hepatitis B virus (genotype B) infection were recruited in this study. 42 patients were treated with interferon-a-1b or a-2b, and 27 patients were treated with PEG interferon (a-2a). Peptide mimicking the C-terminal region of the preS1 protein (94-117aa) of genotype B HBV were synthesised, and the IgG antibody against this peptide was measured by ELISA, and the early response to IFN-alpha therapy was judged by the effect on the viral kinetics, transaminase and the status of HBeAg at 12th week after the treatment. RESULTS: 21 patients were positive for anti-preS1 antibody, and 48 patients were negative for anti-preS1 antibody. After 12 weeks of the treatment, the average decrease in viral levels was 3.37log10 copies/ml and 0.33log10 copies/ml in anti-PreS1 positive patients and anti-preS1 negative patients, respectively, the difference between the two groups was significant (Z = -3.658, P = 0.000); the average decrease in ALT levels was 92 U/L and 30.5 U/L in these two groups, respectively (Z = -2.132, P = 0.033). The rate of hepatitis B e antigen (HBeAg) loss was 41.2% (7/17) and the rate of anti-HBe seroconversion was 5.9% (1/17) in anti-PreS1 positive group, however, the rate of hepatitis B e antigen loss was only 12.8% (5/39), and none of the patients in anti-PreS1 negative group showed anti-HBe seroconversion, the difference between the two groups was significant (Z = -5.110, P = 0.000). The rates of response were 71.4% (15/21) and 16.7% (8/48), respectively, in anti-PreS1 positive group and anti-PreS1 negative group. The rates of complete response were 23.8% (5/21) and 6.25% (3/48), respectively, in these two groups. The positive predictive value (PPV) of anti-C-terminal region of preS1 (94-117aa) antibody in predicting early response was 71.6% and the negative predictive value (NPV) was 83.3%. CONCLUCIONS: Detection of anti-C-terminal region of preS1 (94-117aa) antibody may help to improve the efficacy of INF-alpha therapy for chronic hepatitis B (CHB).