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Objective:To explore genetic etiology and evaluate antifungal immunity in a patient with recurrent cervical lymphadenitis caused by Candida albicans. Methods:Next-generation sequencing was performed to screen susceptibility genes for mycosis in a patient with recurrent cervical lymphadenitis caused by Candida albicans and his parents. Peripheral blood mononuclear cells (PBMCs) and neutrophils were extracted from the patient and 6 healthy controls, and subjected to in vitro co-culture with Candida albicans. Western blot analysis was performed to determine the expression of caspase recruitment domain-containing protein 9 (CARD9) in PBMCs of the patient, enzyme-linked immunosorbent assay to detect levels of tumor necrosis factor-α (TNF-α), interleukin (IL) -6, IL-17A, IL-1β and granulocyte-macrophage colony-stimulating factor (GM-CSF) in the co-culture medium, and a colony-counting method was used to detect the survival rate of Candida albicans after treatment with neutrophils. Statistical analysis was carried out by using t test for comparisons between two groups. Results:Two compound heterozygous mutations were identified in the CARD9 gene of the patient, including c.68C>A (p.S23X) in exon 2 inherited from his father and c.820dupG (p.D274Gfs*61) in exon 6 inherited from his mother. Western blot analysis showed that the relative expression level of CARD9 protein in the PBMCs was 0.41 ± 0.07 in the healthy control group, but CARD9 expression was absent in the patient. After stimulation with heat-inactivated Candida albicans spores, the levels of TNF-α, IL-6, IL-17A, IL-1β and GM-CSF secreted by PBMCs of the patient were significantly lower than those by PBMCs of the healthy controls (all P < 0.001). After 30- and 120-minute in vitro co-culture with neutrophils, the survival rates of Candida albicans were significantly higher in the patient (78.00%, 74.00%, respectively) than in the healthy controls (70.91% ± 1.75%, 34.55% ± 5.35%, t = 3.74, 6.99, respectively, both P < 0.05) . Conclusion:Compound heterozygous mutations were identified in the CARD9 gene of the patient with recurrent cervical lymphadenitis caused by Candida albicans, which led to the absence of CARD9 protein expression, and the patient had a defect in the immunity against Candida albicans.
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Objective:To investigate clinical characteristics of acquired palmoplantar keratodermas (PPK) and the effect of metal implants (MIs) on plasma cytokines in patients with PPK.Methods:An observational study was conducted. Patients with acquired PPK were collected from Department of Dermatology, General Hospital of Eastern Theater Command from June 2020 to December 2021, and health checkup examinees during the same period served as controls. The PPK area and severity index was evaluated according to the eczema area and severity index, metal allergens were detected by metal patch test (MPT) , and plasma levels of cytokines, including tumor necrosis factor (TNF) -α, interferon (IFN) -γ, interleukin (IL) -4, IL-13, IL-17A and IL-8, were detected by enzyme-linked immunosorbent assay. Non-normally distributed measurement data were expressed as median (quartile 1, quartile 3) , and analyzed by rank sum test; intergroup comparisons of enumeration data were performed by chi-square test.Results:A total of 81 PPK patients were collected, including 42 males and 39 females, and their average age was 48 years (range, 21 - 65 years) . In vivo MIs were found in 37 (45.7%) patients, including dental implants in 34, orthopedic implants in 2, and cardiac implants in 1. During the same period, 36 healthy subjects were included in the control group, and there was no significant difference in the age and gender composition between the two groups ( P > 0.05) . No significant difference was observed in the PPK area and severity index between the MI group (37 cases, 0.40 [0.27, 0.75] points) and non-MI group (44 cases, 0.38 [0.19, 0.70] points; Z = 1.21, P = 0.225) . Forty-six patients in the PPK patient group and 30 in the control group were subjected to MPT, and the positive rate of MPT was significantly higher in the patient group (14/46) than in the control group (2/30, χ2 = 6.17, P = 0.013) , and was significantly higher in the patients with in vivo MIs (10/25) than in the patients without (4/21, adjusted P < 0.017) . Plasma cytokines were detected in 36 cases in the patient group and 36 in the control group. The plasma levels of TNF-α and IFN-γ were significantly lower in the patient group (66.2 [58.7, 69.3] pg/ml, 645.0 [571.5, 681.1] pg/ml, respectively) than in the control group (71.5 [64.5, 73.9] pg/ml, 716.5 [620.4, 785.0] pg/ml, respectively; both P < 0.05) , but there was no significant difference in the levels of plasma IL-4, IL-13, IL-17A and IL-8 between the two groups (all P > 0.05) . Conclusions:Positive MPT reaction is more common in acquired PPK patients, especially in patients with in vivo MIs. There may be a correlation between metal and acquired PPK, and changes in plasma TNF-α and IFN-γ levels may be related to the onset of some acquired PPK.
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Current evidence shows that subcutaneous injection of methotrexate results in more rapid and better absorption, higher blood drug levels and less variable exposure than oral dosing. Among patients with rheumatoid arthritis or psoriasis, subcutaneous methotrexate is more effective than oral methotrexate, and has a lower risk of gastrointestinal complaints. For patients in whom oral methotrexate is proved to be inadequate or intolerant, subcutaneous methotrexate also shows good efficacy. Subcutaneous methotrexate can serve as a treatment option before the use of biological agents, and in this way, large amounts of money can be saved. Subcutaneous methotrexate has good application prospects for clinical practice.
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Objective Trichophyton rubrum strains can cause superficial infection and also deep tissue infection, but relevant animal model has not been reported yet.The aim of this study was to construct an animal model of granuloma infected by T.rubrum. Methods Three T.rubrum strains isolated from clinical granuloma tissues, 2 T.rubrum strains isolated from tinea infection and a standard strain of ATCCMYA4438 were selected.Corticosteroids were given to the Balb/C mice before and after the injection of the T. rubrum and mucin suspension and the mice model of granuloma was established.Direct microcopy, culture and histopathologic method were adopt to verify the infection effects. Results The mice inoculated with the T.rubrum granuloma strains with mucin suspension were examined after 21 days in the condition of applying appropriate dose of glucocorticoids.Direct microscopic examination showed the slender mycelium, fungal culture showed the growth of colony and histopathology showed excessive keratinization of foot tissue, formation of granuloma in the dermis with inflammatory cell infiltration of neutro-philic granulocyte and lymphocytes.However, the mice inoculated with the T.Rubrum tinea strains with mucin suspension showed the negative result. Conclusion The rubrum granuloma mice model can be es-tablished using the clinical isolates of T.rubrum granuloma strains with the mucin and glucocorticoids interventions.
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A 76?year?old female patient complained of right chest pain for three months. CT scan showed a clump?like high?density shadow measuring 4.8 cm × 3.0 cm in size in the dorsal portion of the right lower lobe of the lung. Aspiration biopsy was performed, and biopsy samples were subjected to fungal culture and histopathological examination. Histopathological examination showed chronic granulomatous inflammation with hyaline septate hyphae. After 4?day culture, white villous dense colonies were formed on the Sabouraud′s agar medium. The center of the colonies was slightly elevated with wrinkles or radiating striae on the surface, and the bottom of the colonies was faint yellow in color. Microculture yielded abundant septate branched hyphae, and very few colorless hyaline quasi?circular spores. DNA sequencing of rDNA internal transcribed spacer (ITS) regions and β?tubulin genes was performed to identify the isolate, and antifungal susceptibility testing was carried out in vitro. The MEGA7.0 software was used to build phylogenetic trees of Aspergillus fumigatus complex and its closely related species. The isolate was identified as Aspergillus fumigatus by molecular biologic sequencing. The patient was diagnosed with pulmonary aspergilloma. After administration of itraconazole oral solution and vorionazole tablets, the condition got better obviously.
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Eccrine spiradenoma usually occurs as solitary tender nodules. However, it rarely occurs as multifocal localized tumors or has zosteriform distribution. We report the case of a 32-year-old woman with a 20-year history of itchy lesions on the left side of the back and forearm with a zosteriform distribution. Before disease onset, almost no patients have inducing factors, but our patient received an injection beforehand; however, whether this was coincidental or causative remains unknown. The lesions became very itchy after perspiration or eating spicy food, which has never been reported. A literature search revealed 22 cases of multiple segmental eccrine spiradenoma; we summarized the clinical characteristic in order to aid diagnosis and treatment selection.
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Adult , Female , Humans , Diagnosis , Eating , ForearmABSTRACT
Objective Little is known about the role of low oxygen concentration in the formation of the capsule of Cryptococ -cus neoformans .This study was to investigate the effect of hypoxic condition on the capsule size of Cryptococcus neoformans . Methods We cultured 39 Cryptococcus neoformans strains in liquid Sabouraud medium and then incubated them at the normoxic (21%) and hypoxic concentration (1%), respectively.After 72 hours of incubation, we observed the capsules formed at different ox-ygen conditions under the Olympus microscope and the measured their sizes using the Axiovision software . Results Of the total number of Cryptococcus neoformans strains , 30 showed larger capsules in the normoxic than in the hypoxic condition , 17 with statisti-cally significant differences in their diameters (P<0.05), while the other 9 strains exhibited just the opposite results , 4 with statisti-cally significant differences in their diameters ( P<0.05). Conclusion Hypoxia might inhibit the growth of the Cryptococcus neoformans capsule.However, further studies are needed on its correlation with the serum or gene types of the Cryptococcus neofor-mans strain.
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Objective The article was to observe the clinical efficacy of the application of terbinafine and mizolastine in com -bined treatment of chronic eczema ( CE) with dermatophytes infection , so as to define the etiology role of dermatophytes in allergic dis-eases. Methods All subjects were randomly divided into experiment group and control group .The experiment group was treated with the combination of terbinafine and mizolastine , while the control group took mizolastine orally alone .EASI grading , recovery rate and effective rate were evaluated at 2, 3 week after the treatment and EASI grading and recurrence rate were evaluated at 4 weeks after the treatment. Results 79 patients had finished the experiment .Significant difference was found in the effective rates between two groups at 3 weeks after treatment (P<0.05).At 4 weeks after the treatment, EASI value and recurrence rate in experiment group were obviously lower than those in control group (P<0.05). Conclusion Good therapeutic effect has been achieved through the ap-plication of terbinafine and mizolastine in combined treatment of CE with dermatophytes infection , which implies dermatophytes plays an important role in the etiology of CE .
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In recent years , fungal biofilms related infection have become an increasingly important clinical problem .Many clinically important fungi can form biofilms , including:Candida albicans , Cryptococcus neoformans , Rhodotorula species , Aspergillus fumigatus, Malassezia pachydermatis, Histoplasma capsulatum.Many chronic persistent infections are associated with the fungal bio-film formation.The developmental phases of fungal biofilms are complicated , including adhesion, colonisation, maturation and dispers-al, which are governed by many genes .This review discusses clinical impact of biofilm formation of the common human pathogenic fun -gi, biofilm structure and the molecular mechanisms during the biofilm formation process .
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Objective There have been a few reports on chronically recurrent and generalized superficial mycosis caused by trichophyton rubrum.This article was to investigate the cause, diagnosis and therapy of the mycosis. Methods 5 patients with chron-ically recurrent and generalized superficial mycosis caused by trichophyton rubrum were collected from June 2012 to June 2014 in our hospital.Bacterioscopic examination and cultivation were made on skin lesions of the patients.A typical patient who had 7-year course of desease with toenails seriously infected and widespread skin eruption was selected for histopathology examination on skin lesions, mi-crobiology and molecular biology study on 4 bacterial strains isolated from skin lesions in different parts, and in vitro chemosensitivity assay for drug selection.PCR (rDNA ITS sequence analysis) was performed for diagnosis and early treatment. Results Microscopic examintion on skin lesions demonstrated numerous septate, branched hyphae.Cultivation and molecular biology study identified tricho-phyton rubrum.The strain was identified as trichophyton rubrum by ITS sequence analysis and the isolated strains from different lesions were the same fungal species.Histopathology examination revealed slight hyperplasia of squamous epithelium , epidermal hyperkeratini-zation and the upper dermis presented a sparse perivascular lymphocytic infiltrate.The PAS-stain confirmed the presence of few hyphae in the horny layer.The pathogen of this case was trichophyton rubrum. A combination therapy with systemic itraconazole and topically applied terbinafine hydrochloride cream was successful.A follow-up examina-tion one year later showed no recurrence of symptoms. Conclusion The isolation and identification of pathogen is the key to the diagnosis of chronically recurrent and generalized superficial mycosis, with ad-ditional attention to all or none toenail infection.The therapy should not focus simply on the tinea corporis, while comprehensive treatment combined with chemosensitivity assay is preferred.
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Objective There is a lack of information on the effect of hypoxia on the virulence of Cryptococcus neoformans . This study was to construct a Cryptococcus neoformans bilfilm model in vitro and observe the influence of hypoxia on the biofilm forma-tion. Methods We constructed a Cryptococcus neoformans biofilm model in vitro in 24-well and 96-well microculture plates using DMEM culture medium at the oxygen concentrations of 21%( normoxia ) and 1% ( hypoxia ) .We collected the Cryptococcus neofor-mans biofilms at 2, 4, 8, 12, 24, 48, and 72 hours after culturing and observed their thickness , structure, and growth activity in the two different oxygen conditions by light microscopy , confocal laser scanning microscopy , and MTT assay . Results The Cryptococcus neoformans biofilm model was successfully constructed in the conditions of both hypoxia and normoxia .The processes of biofilm forma-tion in the two conditions were similar , involving adhesion , aggregation , micro-colony formation , and biofilm maturation , with the ulti-mate biofilm thickness of about 16 μm.The cell density and growth activity of the biofilms increased with the extension of incubation time, gradually stabilized with their maturity , and both were relatively higher at 1%than at 21%oxygen concentration . Conclusion The abilities of Cryptococcus neoformans biofilm formation vary with different oxygen concentrations , and hypoxia can promote the for-mation of Cryptococcus neoformans biofilms .
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Objective Candidas rank as the second factor of deep fungal infection .This study aimed to investigate the rela-tionship between Candidas and melanin by screening clinically isolated melanin -producing Candidas . Methods We incubated 360 strains of clinically isolated Candidas with caffeic acid , DOPA agar , and minimal medium at a temperature of 35℃, with Cryptococcus neoformans as the positive control .We observed the changes in the culture media using transmission electron microscopy (TEM) and electron spin resonance ( ESR) spectroscopy . Results Cryptococcus neoforman produced pigmentation in all the three types of medi -um.At 5 days, a standard strain of Candida albicans SC 5314 turned dark brown in the DOPA agar and minimal medium and , at 10 days, 12 and 15 of the 360 Candida strains turned dark brown in the DOPA agar and minimal medium , respectively, which were not shown to be melanin by TEM and ESR spectroscopy . Conclusion Culture media are not reliable enough for the identification of mel-anin-producing Candidas , and the observations need to be further verified by TEM and ESR spectroscopy .
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A 48-year-old female presented with a 6-year history of papules and plaques all over the body and with 1-year history of blurred vision in the right eye.Physical examination showed porcelain-white atrophic papules with peripheral erythematous halo and telangiectnsia.She also suffered from exotropia,visual deterioration,visual field defects of the right eye,as well as numbness of the left index finger,thumb and right anterior tibia.Skin biopsies of abdominal lesions revealed dermal necrosis with mucoid degeneration,inflammatory infiltration predominated by lymphocytes around several small blood vessels and occlusion of some blood vessels in deep dermis.Colonoscopy of the whole colon demonstrated scattered patches of hyperemia and erosions with the formation of shallow ulcers.Nerve electromyologram revealed damage to the nerves of right quadriceps femoris muscles.Fecal analysis showed that occult blood was strongly positive.A diagnosis of malignant atrophic papulosis was made based on the characteristic clinical presentation,laboratory and histopathological findings.She was treated with dipyridamole and aspirin for three months,which resulted in no clinical improvement or deterioration.