OBJECTIVES: Verrucous epidermal nevus (VEN), seborrheic keratosis (SK), verruca plana (VP), verruca vulgaris (VV), and nevus sebaceous (NS) are common verrucous proliferative skin diseases with similar clinical appearances, often posing diagnostic challenges. Dermoscopy and reflectance confocal microscopy (RCM) can aid in their differentiation, yet their specific features under these tools have not been systematically described. This study aims to summarize and analyze the dermoscopic and RCM features of VEN, SK, VP, VV, and NS. METHODS: A total of 121 patients with histopathologically confirmed verrucous proliferative skin diseases were enrolled. Dermoscopy and RCM imaging was used to observe and analyze the microscopic features of these conditions. RESULTS: Under dermoscopy, the 5 diseases displayed distinct characteristics: VEN typically showed gyriform structures; SK was characterized by gyriform structures, comedo-like openings, and milia-like cysts; VP and VV featured dotted vessels and frogspawn-like structures; NS presented as brownish-yellow globules. RCM revealed shared features such as hyperkeratosis and acanthosis across all 5 diseases. Specific features included gyriform structures and elongated rete ridges in VEN; pseudocysts and gyriform structures in SK; evenly distributed ring-like structures in VP; vacuolated cells and papillomatous proliferation in VV; and frogspawn-like structures in NS. CONCLUSIONS These 5 verrucous proliferative skin conditions exhibit distinguishable features under both dermoscopy and RCM. The combination of these 2 noninvasive imaging modalities holds significant clinical value for the differential diagnosis of verrucous proliferative skin diseases.
Humans ; Dermoscopy/methods* ; Diagnosis, Differential ; Microscopy, Confocal/methods* ; Male ; Female ; Adult ; Middle Aged ; Adolescent ; Keratosis, Seborrheic/pathology* ; Young Adult ; Warts/diagnosis* ; Child ; Aged ; Skin Diseases/pathology* ; Nevus, Sebaceous of Jadassohn/diagnosis* ; Skin Neoplasms/diagnosis* ; Child, Preschool

OBJECTIVES: Papular dermatoses commonly affecting the female vulva, such as molluscum contagiosum, syringoma, lymphangioma, folliculitis, verruca vulgaris, ectopic sebaceous glands, and bowenoid papulosis, often present with similar clinical appearances and are frequently misdiagnosed. This study aims to explore the clinical diagnostic value of reflectance confocal microscopy (RCM) in differentiating these conditions. METHODS: A retrospective analysis was conducted on RCM imaging and histopathological findings from lesion sites in 172 female patients with vulval papular dermatoses. RCM characteristics confirmed by biopsy were summarized and diagnostic clues were explored. RESULTS: RCM diagnosis was consistent with histopathological diagnosis in 147 out of 172 cases (85.47%). Molluscum contagiosum, syringoma, lymphangioma, and folliculitis all exhibited cystic-like structures under RCM, differing in the location of the structures, wall characteristics, internal contents, and reflectivity. Verruca vulgaris, ectopic sebaceous glands, and bowenoid papulosis lacked such structures. Verruca vulgaris showed distinctive low-refractive vacuolated cells in the spinous layer; bowenoid papulosis exhibited mild cytologic atypia in the spinous layer; ectopic sebaceous glands were characterized by moderately to low-refractive, fish roe-like sebaceous lobules within the dermis. CONCLUSIONS RCM enables noninvasive, real-time, and dynamic visualization of key diagnostic and differential features of common vulvar papular dermatoses in women, offering high diagnostic value.
Humans ; Female ; Microscopy, Confocal/methods* ; Retrospective Studies ; Adult ; Vulvar Diseases/diagnosis* ; Middle Aged ; Young Adult ; Aged ; Adolescent ; Diagnosis, Differential ; Child ; Skin Diseases/pathology* ; Molluscum Contagiosum/diagnosis*

Multilayer paper-based cell culture, as an in vitro three-dimensional (3D) cell culture method, has been frequently used to research drug bioavailability, therapeutic efficacy, and dose-limiting toxicity in malignant tumors. This paper proposes a heterogenous multilayer paper stacking co-culture system to establish a model of natural killer (NK) cells moving through the endothelium layer and attacking tumor spheroids. This system consists of three layers: a bottom tumor-spheroid layer, a middle invasion layer, and a top endothelium layer. NK-92 cells were placed in the supernatant on top of the three layers. After two days of co-culture, the attack of tumor spheroids by NK cells was observed. We additionally examined the infiltration of NK-92 cells within the tumor spheroids at different Z-axis depths using a confocal microscope, and the results suggested that the system successfully realizes NK cells traveling cross the endothelium layer to form tumor-infiltrating NK cells (TINKs). The potential application of multilayer paper for co-culture models involving cancer cells and immune cells holds great promise for exploring the interaction dynamics of these two cell types.
Killer Cells, Natural/immunology* ; Coculture Techniques ; Spheroids, Cellular ; Humans ; Paper ; Cell Line, Tumor ; Microscopy, Confocal

BACKGROUND: Corneal confocal microscopy (CCM) is a noninvasive technique to detect early nerve damage of diabetic sensorimotor polyneuropathy (DSPN). Time in range (TIR) is an emerging metric of glycemic control which was reported to be associated with diabetic complications. We sought to explore the relationship between TIR and corneal nerve parameters in asymptomatic patients with type 2 diabetes (T2DM). METHODS: In this cross-sectional study, 206 asymptomatic inpatients with T2DM were recruited. After 7 days of continuous glucose monitoring, the TIR was calculated as the percentage of time in the glucose range of 3.9 to 10.0 mmol/L. CCM was performed to determine corneal nerve fiber density, corneal nerve branch density, and corneal nerve fiber length (CNFL). Abnormal CNFL was defined as ≤15.30 mm/mm 2 . RESULTS: Abnormal CNFL was found in 30.6% (63/206) of asymptomatic subjects. Linear regression analyses revealed that TIR was positively correlated with CCM parameters both in the crude and adjusted models (all P   <  0.05). Each 10% increase in TIR was associated with a 28.2% (95% CI: 0.595-0.866, P  = 0.001) decreased risk of abnormal CNFL after adjusting for covariates. With the increase of TIR quartiles, corneal nerve fiber parameters increased significantly (all P for trend <0.01). The receiver operating characteristic curve indicated that the optimal cutoff point of TIR was 77.5% for predicting abnormal CNFL in asymptomatic patients. CONCLUSIONS There is a significant independent correlation between TIR and corneal nerve fiber loss in asymptomatic T2DM patients. TIR may be a useful surrogate marker for early diagnosis of DSPN.
Humans ; Diabetes Mellitus, Type 2/complications* ; Cross-Sectional Studies ; Blood Glucose Self-Monitoring ; Blood Glucose ; Nerve Fibers ; Diabetic Neuropathies ; Cornea ; Microscopy, Confocal/methods*

Blood Glucose Self-Monitoring ; Blood Glucose ; Cornea ; Microscopy, Confocal

Confocal laser endomicroscopy technology can obtain cell-level images in real time and in situ, which can assist doctors in real-time intraoperative diagnosis, but its non-invasiveness makes it difficult to relocate the optical biopsy site. The confocal probe localization algorithm can automatically calculate the coordinates of the probe tip, that is, the coordinates of the optical biopsy site. In this paper, a confocal probe localization algorithm based on region growing and endoscope size prior was proposed. The algorithm detected the probe region by region growing on the probe edge image, then searched for tip points based on a given probe axis, and iteratively optimized it. Finally, based on the single-degree-of-freedom motion characteristics of the probe, the three-dimensional coordinates of the tip of the probe were calculated by using the prior information of the size of the endoscope, which solved the scale uncertainty problem of the monocular camera. The confocal probe localization algorithm was tested on the dataset collected in this paper. The results showed that our algorithm no longer relied on the color information of the probe, avoided the influence of uneven illumination on the gray value of the probe pixels, and had a more robust location accuracy and running speed. Within the length of the probe extending out of the endoscope from 0 to 5 cm, the pixel error could be as low as 11.76 pixels, and the average relative position error could be as low as 1.66 mm, which can achieve the real-time and accurate localization of the confocal probe.
Endoscopes ; Algorithms ; Microscopy, Confocal/methods*

Diabetic peripheral neuropathy(DPN),a chronic diabetic microvascular complication with a high incidence among diabetic patients,increases the risk of diabetic foot and amputation.Many methods are available for screening and evaluating DPN,including traditional 10 g monofilament,tuning fork and vibration perception,and tendon reflex tests,which should be combined with some nerve function score systems to improve the detection rate and accuracy for DPN.In recent years,a number of noninvasive new techniques have been developed for the evaluation of nerve injury,such as corneal confocal microscopy,quantitative sensory testing,current perception threshold test,sympathetic sudomotor function evaluation,and quantitative detection of skin advanced glycation end products.This paper reviews these noninvasive methods for screening and evaluating DPN to help clinicians detect and focus on DPN early.
Cornea ; Diabetes Mellitus ; Diabetic Foot ; Diabetic Neuropathies/diagnosis* ; Humans ; Mass Screening ; Microscopy, Confocal

Dermoscopy ; Diagnosis, Differential ; Humans ; Microscopy, Confocal ; Psoriasis/diagnostic imaging* ; Skin Neoplasms/diagnosis*

BACKGROUND: Silica particles (SPs) induce cell proliferation and osteogenic differentiation. We reported that SPs in the scaffold induced early stage osteogenic differentiation. METHODS: A polycaprolactone (PCL) scaffold was fabricated with a 10 wt% SPs. The surface of PCL scaffold was coated with a 10 µg/mL collagen solution. Next, the scaffold was conjugated with 2 µM SPs, 2 µg/mL bone morphogenetic protein 2 (BMP2), or 2 µM BMP2-conjugated SPs (BCSPs). Green fluorescent protein-coupled BMP2 was applied to fabricate the scaffold. The fluorescence intensity was analyzed by confocal microscopy. The mRNA levels of the early osteogenic differentiation marker, alkaline phosphatase (ALP), were analyzed by real-time quantitative polymerase chain reaction. Levels of BMP2, RUNX2, ERK1/2, and AKT were assessed by western blotting. RESULTS: ALP mRNA levels were significantly higher in the BCSP-conjugated scaffold than in the other scaffolds. In the early stage of osteogenic differentiation, the protein levels of BMP2, RUNX2, ERK1/2, and AKT in cells were significantly higher in the BCSP-conjugated scaffold than in other scaffolds. Thus, the BCSP composite scaffold induced rapid osteogenic differentiation. CONCLUSION: These results suggest that BCSP composite can be used to promote early stage osteogenic differentiation and show promise as a material for use in scaffolds for bone regeneration.
Alkaline Phosphatase ; Blotting, Western ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; Bone Regeneration ; Cell Proliferation ; Collagen ; Fluorescence ; Microscopy, Confocal ; Polymerase Chain Reaction ; RNA, Messenger ; Silicon Dioxide ; Stem Cells

PURPOSE: This study aimed to explore the effects and mechanisms of long non-coding RNA (lncRNA) anti-differentiation non-coding RNA (ANCR) on the osteogenesis of osteoblast cells in postmenopausal osteoporosis (PMOP). MATERIALS AND METHODS: Mice models of PMOP were established. ANCR expression and intracellular calcium ions were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and laser confocal microscopy, respectively. ANCR was silenced in osteoblast cells from PMOP mice by the transfection of siRNA-ANCR (si-ANCR). The proliferation and apoptosis of osteoblast cells was analyzed by MTT and flow cytometry, respectively. Alkaline phosphatase (ALP) activity and calcium nodules were examined by ALP and alizarin red staining assay, respectively. The expression of enhancer of zeste homolog 2 (EZH2), runt related transcription factor 2 (RUNX2), and OSTERIX was detected by qRT-PCR and Western blot. Furthermore, an osteogenesis model was constructed in mice, and osteoid formation was observed by hematoxylin-eosin (HE) staining. The interaction between lncRNA-ANCR and EZH2 was further identified by RNA pull-down assay. RESULTS: ANCR expression and intracellular calcium ions were increased in PMOP mice. Si-ANCR significantly increased the proliferation, ALP activity, calcium deposition of osteoblast cells and decreased apoptosis. ANCR and EZH2 were down-regulated by si-ANCR, while RUNX2 and OSTERIX were upregulated. Si-ANCR also promoted osteoid formation in mice treated with hydroxyapatite-tricalcium phosphate. In addition, ANCR specifically bound to EZH2. CONCLUSION: Silencing ANCR promotes the osteogenesis of PMOP osteoblast cells. The specific binding of ANCR with EZH2 suppressed RUNX2, thereby inhibiting osteogenesis.
Alkaline Phosphatase ; Animals ; Apoptosis ; Blotting, Western ; Calcium ; Female ; Flow Cytometry ; Humans ; Ions ; Mice ; Microscopy, Confocal ; Osteoblasts ; Osteogenesis ; Osteoporosis, Postmenopausal ; Real-Time Polymerase Chain Reaction ; RNA ; RNA, Long Noncoding ; RNA, Small Interfering ; RNA, Untranslated ; Transcription Factors ; Transfection