Patients' Knowledge About the Endoscopic UltrasoundGuided Fine Needle Aspiration of Pancreas Is Not Enough.

BACKGROUND: The aim of the study is to evaluate the extent to which patients acquired necessary knowledge about pancreatic endoscopic ultrasound-guided fine needle aspiration and assess what should be more focused on in the informed consent process. METHODS: Adult patients enrolled in this study had pancreatic lesions confirmed by regular imaging and planned to undergo the first pancreatic endoscopic ultrasound-guided fine needle aspiration. These patients were asked to complete a questionnaire, including indications, possible results, downstream events, the risk for false-negative and malignant lesions, and so on. Then we conducted a longterm follow-up of these patients to obtain the final results. RESULTS: Most people (94.25%) correctly recognized that the indication of pancreatic endoscopic ultrasound-guided fine needle aspiration was to exclude malignant lesions. Almost all patients knew that the results could be benign or malignant, while the number of people who were aware of non-diagnostic (22%), indeterminate (18%) outcomes, and the possibility of further testing (20%) after the endoscopic ultrasound-guided fine needle aspiration has decreased significantly. Finally, we got that the false-negative rate and percentages of malignancy were 17.81% and 83.91%, while 98% of participants did not recognize that there is a false-negative risk of endoscopic ultrasound-guided fine needle aspiration and more than 2/3 of participants did not know how much risk they might have for malignant lesions. CONCLUSIONS: A high proportion of patients who received endoscopic ultrasound-guided fine needle aspiration could identify the indication for this procedure but remained unaware of possible outcomes, downstream events, especially the risk for false-negative and malignant lesions. It is necessary to improve the quality of dialogue between clinicians and patients, and the information about the risk of false-negative and malignancy may need to be emphasized in the informed consent process.

Talaromyces marneffei Influences Macrophage Polarization and Sterilization Ability via the Arginine Metabolism Pathway in Vitro.

The opportunistic fungal pathogen Talaromyces marneffei, which is endemic across a narrow band of tropical Southeast Asia and southern China, is an intracellular pathogen that causes systemic and lethal infection through the mononuclear phagocyte system. The mechanisms by which T. marneffei successfully replicates and escapes the immune system remain unclear. To investigate the role of arginine metabolism in the escape of T. marneffei from killer macrophages, we assessed inducible nitric oxide synthase (iNOS) and arginase expression, nitric oxide (NO) production, arginase and phagocytic activity, and the killing of T. marneffei in a coculture system. Our results indicate that T. marneffei induced macrophage polarization toward the M2 phenotype and regulated the arginine metabolism pathway by prolonging infection, thereby reducing antimicrobial activity and promoting fungal survival. Moreover, inhibiting T. marneffei-induced macrophage arginase activity with Nω-hydroxy-nor-arginine restored NO synthesis and strengthened fungal killing. These findings indicate that T. marneffei affects macrophage polarization and inhibits macrophage antimicrobial function via the arginine metabolism pathway.

Overexpression of CD86 enhances the ability of THP-1 macrophages to defend against Talaromyces marneffei.

BACKGROUND: Macrophages are the first line of defense against Talaromyces marneffei. CD86 is a surface molecule expressed on antigen-presenting cells, such as macrophages, that provide costimulatory signals necessary for T cell activation and survival. In a prior study, it was shown that as infection progressed, CD86 expression levels in macrophages considerably declined while CD86 concentrations in the supernatant significantly increased. Additionally, M1 macrophage polarization was insufficient and switched to M2 macrophage polarization. Besides costimulation, however, additional roles of CD86 are not known or well-studies. Therefore, we hypothesized that upregulating CD86 on macrophages might promote T. marneffei defense. METHODS: A lentivirus vector, called Lenti-CD86, was used to infect THP-1 cells to overexpress secretory CD86. Through killing assay, nitric oxide detection, and cytokine detection, the capacity of THP-1 macrophages to phagocytose and kill T. marneffei was examined. RESULTS: In the current study, Lenti-CD86 transfection of THP-1 cells resulted in a signifant expression of CD86. Additionally, the THP-1 macrophages stably transfected with Lenti-CD86 showed higher nitric oxide and IL-1ß production, faster polarization, and stronger phagocytosis and killing capabilities than the non-transfected or control virus transfected cells. CONCLUSION: Our study shows that lentivirus-mediated CD86 overexpression improves THP-1 macrophages' capacity to phagocytose and eliminate T. marneffei.

The Effect of Talaromyces marneffei Infection on CD86 Expression in THP-1 Cells.

BACKGROUND: Talaromyces marneffei (T. marneffei) is a destructive opportunistic dimorphic fungal which can cause lethiferous Talaromycosis, but the clearance of T. marneffei mainly depends on the innate immune response. OBJECTIVE: To investigate whether T. marneffei can inhibit the expression of CD86 in THP-1 cells after infection and discuss the potential mechanisms. METHODS: Western blot and immunoelectron microscopy were used to detect the CD86 expression on T. marneffei cultured on BHI medium at 37°C. Western blot, enzyme-linked immunoassay and immunofluorescence were used to detect the change of CD86 expression on macrophages incubating with T. marneffei. Enzyme-linked immunoassay was used to detect the content of CD86 in supernatant in the co-culture system. Immunohistochemistry and immunoelectron microscopy were used to detect the expression of CD86 on T. marneffei incubating with macrophages. RESULTS: T. marneffei did not express CD86 when cultured separately at 37°C detected by Western blot and immunoelectron microscopy, but it did express CD86 when incubated with macrophages detected by immunohistochemistry and immunoelectron microscopy. The CD86 expression of macrophages significantly decreased at 72 hours when infected with T. marneffei while the content of CD86 in supernatant significantly increased at 72 hours compared with the control group which were detected by Western blot, enzyme-linked immunoassay and immunofluorescence. CONCLUSION: 1) After T. marneffei infection, CD86 expression on THP-1 decreased, and with the progression of infection, insufficient polarization of M1 macrophages gradually appeared; 2) T. marneffei may adsorb or uptake CD86 in supernatant produced by macrophages during the contact with THP-1 cells, thus leading to the consumption of CD86 in macrophages.