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 the protective effect and mechanism of isorhamnetin on myocardial injury in rats with acute myocardial infarction (AMI).Methods:The AMI rat model was established by coronary artery left anterior descending ligation. The SD rats were divided into sham-operated group, model group, Fasudil group (30 mg/kg) and low-, medium-, and high-dose (25, 50, 100 mg/kg) groups. Each group had 5 rats. They were administrated intragastric, once a day, and were treated for 14 d. Color Doppler ultrasonography was used to detect cardiac function by measuring left ventricular end-systolic diameter (LVESd), left ventricular end-diastolic diameter (LVEDd), and left ventricular long-axis shortening fraction (FS). Myocardial infarct size was detected by TCT staining. Superoxide dismutase (SOD), malondialdehyde (MDA), interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) levels in rat serum were detected by ELISA and TUNEL assay for the cardiomyocyte apoptosis index. Cysteinyl aspartate specific proteinase-3 (Caspase-3) activity in myocardial tissue was detected by the Caspase-3 activity assay kit, and the expression of Bcl-2 associated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) proteins in myocardial tissue was detected by Western Blot.Results:Compared with the sham operation group, the FS, SOD content, and expression levels of Bcl-2 protein in the model group were significantly decreased (all P<0.05), and LVEDd, LVESd, myocardial infarct size, MDA content, IL-6 content, TNF-α content, IL-1β content, apoptosis index, Caspase-3 activity, and Bax protein expression level were significantly increased (all P<0.05). There was no significant difference between the low concentration of isorhamnetin and the model group (all P>0.05). However, the above changes caused by the construction model after treatment with Fasudil and medium- and high-concentrations of isorhamnetin significantly reversed (all P<0.05). Conclusions:Isorhamnetin can improve cardiac function and protect myocardial injury in AMI rats by reducing oxidative stress and inflammatory damage to cardiomyocytes and inhibiting cardiomyocyte apoptosis.

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.

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 To evaluate the role of caveolin-1 (Cav-1) in penehyclidine hydrochioride (PHC)-induced inhibition of lipopolysaccharide(LPS)-induced activation of Toll-like receptor 4 (TLR4) /p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in macrophages of mice.Methods Macrophages of mice were seeded in 6 em diameter dishes (5 ml per dish) and divided into 5 groups (n=20 each) using a random number table:Scr-siRNA group (S group),Scr-siRNA + LPS group (LPS group),Ser-siRNA+LPS +PHC group (LPS+P group),Cav-1-siRNA+LPS group (C+LPS group) and Cav-1-siRNA+LPS+PHC group (C+LPS+P group).Macrophages were transfected with Scr-siRNA for 24 h in S,LPS and LPS+P groups and with Smart pool Cav-1 siRNAs for 24 h in C+LPS and C+LPS+P groups.LPS at the final concentration of 1 μg/ml was added after the end of transfection,and macrophages were then incubated for 2 h in LPS,LPS+P,C+LPS and C+LPS+P groups.In LPS+P and C+LPS+P groups,PHC at the final concentration of 2 μg/ml was added at 2 h of incubation with LPS,and macrophages were then incubated for 2 h.The expression of Cav-1 and TLR4 was detected by Western blot.The expression of p38 MAPK was determined by immunofluorescence.The level of tumor necrosis factor-alpha (TNF-α) in the culture medium was determined by enzyme-linked immunosorbent assay.The activity of myeloperoxidase (MPO) in macrophages was measured by colorimetry.Results Compared with group S,the expression of TLR4 and p38 MAPK was significantly up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in the other four groups,the expression of Cav-1 was significantly downregulated in LPS and C+LPS groups (P ＜0.05),and no significant change was found in the expression of Cav-1 in group LPS+P (P＞0.05).Compared with group LPS,the expression of Cav-1 was significantly up-regulated,the expression of TLR4 and p38 MAPK was down-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were decreased in group LPS+P,and the expression of Cay-1 was significantly down-regulated,the expression of TLR4 and p38 MAPK was up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in group C+LPS (P＜0.05).Compared with group LPS+P,the expression of Cav-1 was significantly down-regulated,the expression of TLR4 and p38 MAPK was up-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were increased in group C+LPS+P (P＜0.05).Compared with group C+LPS,the expression of Cav-1 was significantly up-regulated,the expression of TLR4 and p38 MAPK was down-regulated,and the concentration of TNF-α in the culture medium and activity of MPO were decreased in group C+LPS+P (P＜0.05).Conclusion The mechanism by which PHC inhibits LPS-induced activation of TLR4/p38 MAPK signaling pathway in macrophages is related to up-regulating Cav-1 expression in mice.

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.

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.
Adult ; Diagnosis ; Eating ; Female ; Forearm ; Humans

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 .

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 .