Mechanosensitive TRPV4 is required for crystal-induced inflammation.

Crystal structures activate innate immune cells, especially macrophages and initiate inflammatory responses. We aimed to understand the role of the mechanosensitive TRPV4 channel in crystal-induced inflammation. Real-time RT-PCR, RNAscope in situ hybridisation, and Trpv4eGFP mice were used to examine TRPV4 expression and whole-cell patch-clamp recording and live-cell Ca2+ imaging were used to study TRPV4 function in mouse synovial macrophages and human peripheral blood mononuclear cells (PBMCs). Both genetic deletion and pharmacological inhibition approaches were used to investigate the role of TRPV4 in NLRP3 inflammasome activation induced by diverse crystals in vitro and in mouse models of crystal-induced pain and inflammation in vivo. TRPV4 was functionally expressed by synovial macrophages and human PBMCs and TRPV4 expression was upregulated by stimulation with monosodium urate (MSU) crystals and in human PBMCs from patients with acute gout flares. MSU crystal-induced gouty arthritis were significantly reduced by either genetic ablation or pharmacological inhibition of TRPV4 function. Mechanistically, TRPV4 mediated the activation of NLRP3 inflammasome by diverse crystalline materials but not non-crystalline NLRP3 inflammasome activators, driving the production of inflammatory cytokine interleukin-1ß which elicited TRPV4-dependent inflammatory responses in vivo. Moreover, chemical ablation of the TRPV1-expressing nociceptors significantly attenuated the MSU crystal-induced gouty arthritis. In conclusion, TRPV4 is a common mediator of inflammatory responses induced by diverse crystals through NLRP3 inflammasome activation in macrophages. TRPV4-expressing resident macrophages are critically involved in MSU crystal-induced gouty arthritis. A neuroimmune interaction between the TRPV1-expressing nociceptors and the TRPV4-expressing synovial macrophages contributes to the generation of acute gout flares.

Evaluating the accuracy and diagnostic value of CFW and a new fluorescent reagents, fluorescent brightener 85, for the diagnosis of vulvovaginal candidiasis.

BACKGROUND: Vulvovaginal candidiasis (VVC) is a common vaginitis in females. The commonly used diagnostic method, 10% potassium hydroxide (KOH) smear microscopy, makes it not very easy to recognize fungi. METHODS: Vaginal secretions were collected from clinically suspected VVC patients and divided into four groups and examined using KOH, CFW (Calcofluor White), FB 85(fluorescent brightener 85), and culture. The data were statistically analyzed. RESULTS: In total, 110 patients with suspected VVC were recruited. The positive rates of KOH, CFW, FB 85, and the culture method were 68.2%, 64.5%, 61.8%, and 77%, respectively. According to the McNemar test, there was no statistically significant difference between the KOH, CFW, and the FB 85 methods (p > 0.05). However, CFW had a shorter diagnosis time than the KOH method and had a statistically significant difference (p < 0.001). Moreover, CFW has the highest sensitivity, specificity, and accuracy. In morphological recognition, it was easier to recognize fungal structures with CFW and FB 85 than with the KOH. CONCLUSIONS: The fluorescent method is a good method for the diagnosis of VVC. And the fungi can be found more quickly. Similar to CFW, FB 85 is also a potential good fluorescent reagent for the diagnosis of VVC and has potential value for application in clinical fungal infection diseases.

Application of reflectance confocal microscopy to investigate the non-ablative, micro-ablative, and ablative effects of CO2 fractional laser irradiation on skin.

CO2 fractional laser, as an ablative fractional laser, is commonly used in cosmetic treatment. We applied CO2 fractional laser irradiation to skin in vitro and used reflectance confocal microscopy (RCM) to image and detect the presence of any non-ablative, micro-ablative and ablative effects, in order to better understand the features of a CO2 fractional laser. In vitro irradiation of foreskin was performed using a CO2 fractional laser. Foreskin specimens were divided into 4 groups that received different amounts of irradiation energy, based on the number of irradiation passes they received: 5, 10, 15, and 20 passes, respectively. This corresponds to fluence energy of 16.3, 32.6, 48.9, 65.3 J/cm2. Immediately after irradiation, digital microscopy (DM), RCM, and histopathology were performed to observe whether the non-ablative, micro-ablative, and ablative phenomenon occurred, and the injury features of MTZs. Immediately after CO2 fractional laser irradiation, RCM and DM showed that when the numbers of passes were 5 and 10, a micro-ablative column (MAC) could not be observed or was very small. We mainly observed a thicker thermal coagulation zone (TCZ), representing non-ablative or micro-ablative effects. When the number of passes were increased to 15 and 20, the MAC was significantly enlarged and surrounded by a TCZ of medium thickness, representing ablative effects. For the first time, this study used RCM and DM to demonstrate that a CO2 fractional laser could achieve non-ablative, micro-ablative, and ablative effects on irradiated skin via different energy levels.

Evaluation of a new fluorescent reagent, fluorescent brightener 85, for the diagnosis of suspected onychomycosis compared with potassium hydroxide.

Onychomycosis is a common fungal infection in dermatology clinics. The commonly used diagnostic method, potassium hydroxide (KOH) direct microscopy, does not have a high-positive detection rate. Therefore, a new diagnostic method is needed to confirm onychomycosis. The objective of this research was to use fluorescent brightener 85 (FB 85), a new fluorescent reagent for fungi detection, and to evaluate this new method based on the diagnostic positive rate, accuracy and efficiency in suspected onychomycosis compared with the KOH method. Subungual debris was collected from clinically suspected onychomycosis cases and divided into two parts. The parts were examined by FB 85 and 10% KOH solution, respectively. In total, 108 patients with suspected onychomycosis were recruited. The positive rates of FB 85 and KOH were 88.9% and 55.6%, respectively. The patients showed significant statistical differences (P < .001). Compared with the KOH method, the FB 85 fluorescent method was an accurate and efficient method with a higher positive rate. This method could increase the diagnostic accuracy and efficiency of onychomycosis detection.

Application of reflectance confocal microscopy to evaluate skin damage after irradiation with an yttrium-scandium-gallium-garnet (YSGG) laser.

The objective of this study was to observe the characteristics of the skin after irradiation with a 2790-nm yttrium-scandium-gallium-garnet (YSGG) laser using reflectance confocal microscopy (RCM). A 2790-nm YSGG laser was used to irradiate fresh foreskin (four doses, at spot density 3) in vitro. The characteristics of microscopic ablative columns (MAC), thermal coagulation zone (TCZ), and microscopic treatment zones (MTZ) were observed immediately after irradiation using digital microscope and RCM. The characteristics of MAC, TCZ, and MTZ with variations in pulse energy were comparatively analyzed. After irradiation, MAC, TCZ, and MTZ characteristics and undamaged skin between MTZs can be observed by RCM. The depth and width of MTZ obviously increased with the increase in pulse energy. At 80, 120, and 160 mJ/microbeam (MB), the MTZ actual area and proportion were about two times that of the theoretical value and three times at 200 mJ/MB. With increases in depth, the single MAC gradually decreased in a fingertip-shaped model, with TCZ slowly increasing, and MTZ slightly decreasing in a columnar shape. RCM was able to determine the characteristics of thermal injury on the skin after the 2790-nm YSGG laser irradiation with different pulse energies. Pulse energy higher than 200 mJ/MB may have much larger thermal injury and side effect. RCM could be used in the clinic in future.

An ultrastructural study of Trichophyton rubrum induced onychomycosis.

BACKGROUND: Trichophyton rubrum (T.rubrum) caused onychomycosis is the most common nail fungal disease. The common diagnostic methods are direct microscopic examination and fungal culture. In this study we used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to study the subungual ultrastructural changes in T. rubrum induced onychomycosis. METHODS: Six outpatients with onychomycosis were recruited and T.rubrum infection was confirmed by fungal culture. Six toenail samples were collected and prepared for SEM characterization. The cultured fugal colonies were prepared for SEM and TEM characterization. RESULTS: 1) SEM showed significant structural damages and the formation of a thin layer or a single layer of keratinocytes in all infected nail plates. Hyphae (piercing or penetrating keratinocytes layers), arthrospores and local bacterial aggregation were observed on the ventral surface of the nail plates. 2) SEM of the cultured fungal colony showed relatively straight, highly branched hyphae and microconidias; TEM showed branching hyphae that were composed of double-layer cell walls. Hyphae had nucleus, mitochondria, liposomes, lysosomes, scattered rough endoplasmic reticulum, myeloid bodies and aggregated ribosomes. There were high-density particles outside the hyphae. CONCLUSION: SEM showed a large number of hyphae penetrated the keratinocytes layer, suggesting that T. rubrum can cause severe damage to the stratum corneum. TEM showed the ultrastructural features of T. rubrum-induced infection before treatment.

Impact of metformin on melanoma: a meta-analysis and systematic review.

Background: There is evidence of a modest reduction in skin cancer risk among metformin users. However, no studies have further examined the effects of metformin on melanoma survival and safety outcomes. This study aimed to quantitatively summarize any influence of metformin on the overall survival (OS) and immune-related adverse effects (irAEs) in melanoma patients. Methods: Selection criteria: The inclusion criteria were designed based on the PICOS principles. Information sources: PubMed, EMBASE, Cochrane Library, and Web of Science were searched for relevant literature published from the inception of these databases until November 2023 using 'Melanoma' and 'Metformin' as keywords. Survival outcomes were OS, progression-free survival (PFS), recurrence-free survival (RFS), and mortality; the safety outcome was irAEs. Risk of bias and data Synthesis: The Cochrane tool for assessing the risk of bias in randomized trial 2 (RoB2) and methodological index for non-randomized studies (MINORS) were selected to assess the risk of bias. The Cochrane Q and I 2 statistics based on Stata 15.1 SE were used to test the heterogeneity among all studies. Funnel plot, Egger regression, and Begg tests were used to evaluate publication bias. The leave-one-out method was selected as the sensitivity analysis tool. Results: A total of 12 studies were included, involving 111,036 melanoma patients. The pooled HR for OS was 0.64 (95% CI [0.42, 1.00], p = 0.004, I2 = 73.7%), HR for PFS was 0.89 (95% CI [0.70, 1.12], p = 0.163, I2 = 41.4%), HR for RFS was 0.62 (95% CI [0.26, 1.48], p = 0.085, I2 = 66.3%), and HR for mortality was 0.53 (95% CI [0.46, 0.63], p = 0.775, I2 = 0.0%). There was no significant difference in irAEs incidence (OR = 1.01; 95% CI [0.42, 2.41]; p = 0.642) between metformin and no metformin groups. Discussion: The improvement in overall survival of melanoma patients with metformin may indirectly result from its diverse biological targets and beneficial effects on multiple systemic diseases. While we could not demonstrate a specific improvement in the survival of melanoma patients, the combined benefits and safety of metformin for patients taking the drug are worthy of recognition. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42024518182.

Large-Scale Proteome Profiling Identifies Biomarkers Associated with Suspected Neurosyphilis Diagnosis.

Neurosyphilis (NS) is a central nervous system (CNS) infection caused by Treponema pallidum (T. pallidum). NS can occur at any stage of syphilis and manifests as a broad spectrum of clinical symptoms. Often referred to as "the great imitator," NS can be easily overlooked or misdiagnosed due to the absence of standard diagnostic tests, potentially leading to severe and irreversible organ dysfunction. In this study, proteomic and machine learning model techniques are used to characterize 223 cerebrospinal fluid (CSF) samples to identify diagnostic markers of NS and provide insights into the underlying mechanisms of the associated inflammatory responses. Three biomarkers (SEMA7A, SERPINA3, and ITIH4) are validated as contributors to NS diagnosis through multicenter verification of an additional 115 CSF samples. We anticipate that the identified biomarkers will become effective tools for assisting in diagnosis of NS. Our insights into NS pathogenesis in brain tissue may inform therapeutic strategies and drug discoveries for NS patients.

Importance of controlling mesocotyl elongation in the development of rice seedlings intended for mechanical transplantation.

The mesocotyl facilities the emergence of deep-sown rice. However, the effects of mesocotyl elongation on mechanically transplanted rice seedlings remain unclear. In this study, the indica three-line hybrid rice Chuanyou 6709 (CY6709) and the indica conventional rice Guichao II (GCII) were selected as experimental materials. The seedlings were grouped based on mesocotyl lengths of 1.0 and 2.0 cm (M1 and M2, respectively), and seedlings without mesocotyl elongation were used as a control (M0). Seedling morphology, root morphology and physiology, and dynamic changes in soluble sugar and protein, malondialdehyde (MDA), and antioxidant enzyme activity in the mesocotyl were evaluated. The results showed that the elongation of mesocotyl is not conducive to improving the quality of mechanically transplanted seedlings, resulting in weak seedlings and poor root coiling force. The mesocotyl lengths of the seedlings showed a single peak with increasing seedling age, which gradually disappeared. The longer the mesocotyls, the slower their senescence. The MDA content of M2 was significantly lower than that of M1, and the activities of soluble sugar, soluble protein, and antioxidant enzymes of M2 were higher than those of M1, implying that seedlings with longer mesocotyls yielded lower-quality seedlings, reducing their suitability for mechanized transplantation. Compared with those of M0, the root-shoot ratio, stem base width, leaf age, leaf area, white root number, root coiling force, root length, root surface area, and root volume of M1 and M2 were reduced. Therefore, in the raising of rice seedlings, excessive elongation of the rice mesocotyl is not conducive to optimum root growth and development of aboveground structures for seedlings that are suitable for mechanized transplantation. Controlling the mesocotyl elongation can facilitate the cultivation of high-quality mechanically transplanted seedlings.

Mechanisms of Broad-Band UVB Irradiation‒Induced Itch in Mice.

Although sunburn can produce severe uncontrollable itching, the underlying mechanisms of UV irradiation‒induced itch are poorly understood because of a lack of experimental animal models of sunburn itch. In this study, we established a sunburn-related mouse model and found that broad-band UVB irradiation elicited scratching but not wiping behavior in mice. Using a combination of live-cell calcium ion imaging and quantitative RT-PCR on dorsal root ganglion neurons, H&E staining, immunofluorescence staining of skin preparations, and behavioral testing, in combination with genetic and pharmacological approaches, we showed that TRPV1-positive dorsal root ganglion neurons but not mast cells are involved in broad-band UVB irradiation‒induced itch. Moreover, both genetic and pharmacological inhibition of TRPV1 function significantly alleviated the broad-band UVB irradiation‒induced itch response. Collectively, our results suggest that broad-band UVB irradiation evokes itch sensation in mice by promoting TRPV1 channel function in dorsal root ganglion neurons and provide potential therapeutic targets for sunburn-related itch.

The Role of Scanning Electron Microscopy in the Direct Diagnosis of Onychomycosis.

PURPOSE: The purpose of this study was to evaluate the role of scanning electron microscopy (SEM) in the direct diagnosis of suspected onychomycosis with negative mycological test results. METHODS: Outpatients diagnosed with suspected onychomycosis with negative mycological test results, including direct microscopic examination with 10% potassium hydroxide (KOH) and fungal culture, on 3 separate occasions were recruited. A small piece of infected nail was obtained for SEM examination. RESULTS: Among the 48 suspected onychomycosis samples, SEM revealed that 18 (37.5%) were positive for fungal structures, including 10 (20.8%) cases of hyphae and 8 (16.7%) cases of yeast blastospores or budding. CONCLUSION: SEM represents an effective method to diagnose suspected onychomycosis when the traditional mycological methods were negative. Therefore, this technique could be used in clinical practice.

Scanning electron microscopy of the nail plate in onychomycosis patients with negative fungal culture.

Onychomycosis is a common dermatological problem and can be identified by direct microscopic examination and fungal culture. However, the positive rate of fungal culture is low. This study investigated the application of scanning electron microscopy in the diagnosis of onychomycosis in 20 patients with negative fungal culture. In this study, a routine glutaraldehyde fixation method was used to prepare specimens for electron microscope examination. Results showed that under the scanning electron microscope, significant structural damage was observed in the nail plate in all patients. Hyphaes were seen in 70% of cases. A mixture of scattered hyphaes, pseudohyphaes, and spores was observed in 30% of cases. A mixture of spores and bacteria was observed in 10% of cases. A mixture of hyphaes and bacteria was observed in 20% of cases. The typical hyphae pierced a thin layer or single layer of corneocytes. Hyphaes could be smooth, sleek, and straight with visible separation, or dry, bent, and folded with a smooth surface. The diameter of hyphaes was 1-2 µm. The scattered spores were the main form of spore growth, and the growth of budding spores can be seen attached to the surface of layered armor. Most of the bacteria were gathered in clumps on the ventral surface, especially in grooves. In conclusion, scanning electron microscopy can be used to preliminarily identify the pathogen involved and the degree of damage in cases where onychomycosis is clinically diagnosed, but fungal culture is negative.