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.

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.

Postzygotic mutations are acquired in normal tissues throughout an individual's lifetime and hold clues for identifying mutagenic factors.Here,we investigated postzygotic mutation spectra of healthy individuals using optimized ultra-deep exome sequencing of the time-series samples from the same volunteer as well as the samples from different individuals.In blood,sperm,and muscle cells,we resolved three common types of mutational signatures.Signatures A and B represent clock-like mutational processes,and the polymorphisms of epigenetic regulation genes influence the pro-portion of signature B in mutation profiles.Notably,signature C,characterized by C＞T transitions at GpCpN sites,tends to be a feature of diverse normal tissues.Mutations of this type are likely to occur early during embryonic development,supported by their relatively high allelic frequencies,presence in multiple tissues,and decrease in occurrence with age.Almost none of the public datasets for tumors feature this signature,except for 19.6％of samples of clear cell renal cell carcinoma with increased activation of the hypoxia-inducible factor 1(HIF-1)signaling pathway.Moreover,the accumulation of signature C in the mutation profile was accelerated in a human embryonic stem cell line with drug-induced activation of HIF-1α.Thus,embryonic hypoxia may explain this novel signature across multiple normal tissues.Our study suggests that hypoxic condition in an early stage of embryonic development is a crucial factor inducing C＞T transitions at GpCpN sites;and indi-viduals'genetic background may also influence their postzygotic mutation profiles.

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.

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.

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.

Objective To evaluate the value of MSCT and MRI in diagnosis of early postoperative complications of inguinal hernia repair (IHR).Methods Imaging and clinical data in 1 6 patients with early complications of IHR were analyzed retrospectively.Re-sults Among the complications,postoperative infection in 2 was found including incision infection in 1 and groin cellulitis in other 1. CT showed swelling abdominal wall and heterogeneous enhancement for incision infection,and inguinal mass,deep inguinal ring thickening,edema of residual sac with fluid and air collections for groin cellulitis.Seroma was found in 8,and CT and MRI demon-strated residual sac effusion,spermatic cord thickening and spermatic vascular tortuosity.Residual sac edema was found in 4,and CT and MRI showed thickening spermatic cord without effusion in residual sac or scrotum.Effusions between patch and anterior abdom-inal wall were detected by MRI in 2,one of which was accompanied by patch shrinking.Conclusion With specific clinical background for early complications of IHR,MSCT and MRI provide more anatomical information of inguinal region,which may contribute to di-agnosis and treatment of early postoperative complications.

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 .

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.

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 .