OBJECTIVE: To explore the effects of two methods of smear layer removal on the surface properties of dentin. METHODS: Sixty extracted sound third molars were collected in this study, and were prepared as uniform dentin specimens with smear layer. All specimens were randomly divided into three groups: Control group, ultrasonic treatment (UT) group and etched treatment (ET) group. Scanning electron microscope (SEM) were used to observe the surface micromorphology of all three groups. Then, the surface elements, mineral phases and functional groups were analyzed by energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and flourier transformed infrared spectrometer (FTIR) respectively. The mechanical properties, hydrophilicity and biocompatibility were also further evaluated. RESULTS: It was revealed that dentin tubules of UT and ET groups were exposed, but lots of dentin debris piled up on the surface of the control one which covered up dentin tubules on the surface. The EDX results should that the weaker peak value of calcium and phosphorus in ET group than control and UT groups. Characteristic peaks of hydroxyapatite could be seen by XRD in all of the three groups, but lower distinctive peaks of amide Ⅰ, Ⅱ and Ⅲ bands of collagen of the dentin surface in control group than in ET and UT groups. The microhardness results showed that ET group was lower than control and UT groups, the difference was significant (P < 0.05). Better hydrophilicity of ET group was investigated (P < 0.05) than control group and UT group. Cells could be observed to adhere normally to dentin surface of each group which meant that all of the three groups had good biocompatibility. CONCLUSION Both UT and ET could effectively remove the smear layer on the surface of dentin and had no adverse effect of the dentin micromorphology and biocompatibility. The ultrasonic removal of the smear layer did not influence the mineral structure, hydrophilicity and mechanical properties of dentin surface. Although ET can effectively improve the hydrophilicity of dentin but decreased mechanical properties and the content of calcium and phosphorus.
Dentin/ultrastructure* ; Humans ; Surface Properties ; Smear Layer ; Molar, Third ; Microscopy, Electron, Scanning ; Dental Etching/methods*

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*

OBJECTIVES: We aimed to compare the apical sealing properties of three endodontic sealers, namely, C-Root SP (C-R), iRoot SP, and GuttaFlow Bioseal (GFB) in vitro. METHODS: Eighty-two single-rooted premolars and anterior teeth were prepared by using M3 machine with nickel-titanium file and randomly divided into six experimental groups (n=12) and two control groups (n=5). Group A1: single-cone technique (SC)+C-R; group B1: SC+iRoot SP; group C1: SC+GFB; group A2: single-cone with ultrasonic activation (SU)+C-R; group B2: SU+iRoot SP; group C2: SU +GFB; group D: positive control group, and group E: negative control group. Dye penetration length and lateral root canal filling in each group were measured by dye penetration test. A scanning electron microscope (SEM) was used to observe the interface between gutta pertscha, root canal sealer, and dentin wall. Dye penetration length was measured and analyzed by Kruskal-Wallis test, and data on lateral root canal filling were evaluated using Chi-square. RESULTS: The dye penetration length in group A1 was lower than that in groups C1 and A2 (P<0.05) but was not significantly different from the other groups (P>0.05). Lateral root canal filling was not significantly different among all groups (P>0.05). SEM showed that GFB was slightly better than C-R and iRoot SP in binding to gutta pertcha and dentin wall. CONCLUSIONS GFB, C-R, and iRoot SP demonstrate excellent apical sealing ability. Under the conditions tested in this study, SU did not yield significantly improve the apical sealing ability of the three root canal sealers.
Root Canal Filling Materials/chemistry* ; Humans ; Gutta-Percha ; Microscopy, Electron, Scanning ; Root Canal Obturation/methods* ; Ceramics ; Dimethylpolysiloxanes ; Drug Combinations

OBJECTIVES: This study aimed to compare the effectiveness of ultrasound and acoustic and laser cleaning of curved root canals. METHODS: A total of 92 molars with independent root canals with a curvature of 20°-40° were prepared and standardized at 04 25# and stained with gentian violet solution for 72 h. Among them, 52 were randomly divi-ded into four groups for final rinsing (n=13): NI group, PUI group, EDDY group, and PIPS group. Ten samples in each group were cut horizontally along the long axis perpendicular to the root and divided into curved upper, curved, and apical segments. Images were taken with a stereomicroscope and Image J measurements were taken to calculate the depth of rinse penetration. The remaining three samples from each group were split along the long axis of the dentin, photographed by scanning electron microscope to record the dentin tubule exposure and staining layer, and scored for staining layer by double-blind method. SPSS 26.0 software was used to perform statistical analysis and select the best flushing method. An extra 40 samples were randomly divided into four groups for detection of flushing fluid penetration depth (n=10): 10, 20, 30, and 40 s. RESULTS: In the upper part, the mean depth of infiltration was not significantly different between the experimental and control groups (P>0.05). The PIPS group had a significantly lower smear layer score than the control group and the EDDY group (P<0.01). In the curved segment, the mean depth of infiltration was significantly greater in the PUI group than in the control group (P<0.05); the tarnish layer score was lower in each experimental group than in the control group. At the top, the mean depth of infiltration was greater in the PUI and PIPS groups than in the control group (P<0.05), and the smear layer score was lower in the PIPS group than in the other groups (P<0.05). After the time was changed, the depth of infiltration of PUI increased only in the apical segment as the flushing time increased. CONCLUSIONS The PUI and PIPS methods facilitate the penetration of irrigation solution into the dentin canal in curved root canals, especially in the apical segment. The PIPS technique is effective in removing the smear layer in curved root canals.
Humans ; Dental Pulp Cavity ; Microscopy, Electron, Scanning ; Root Canal Irrigants ; Root Canal Preparation/methods* ; Smear Layer ; Sodium Hypochlorite ; Therapeutic Irrigation/methods* ; Double-Blind Method

Urinary system tumors affect a huge number of individuals, and are frequently recurrent and progressing following surgery, necessitating lifelong surveillance. As a result, early and precise diagnosis of urinary system cancers is important for prevention and therapy. Histopathology is now the golden stan-dard for the diagnosis, but it is invasive, time-consuming, and inconvenient for initial diagnosis and re-gular follow-up assessment. Endoscopy can directly witness the tumor's structure, but intrusive detection is likely to cause harm to the patient's organs, and it is apt to create other hazards in frequently examined patients. Imaging is a valuable non-invasive and quick assessment tool; however, it can be difficult to define the type of lesions and has limited sensitivity for early tumor detection. The conventional approaches for detecting tumors have their own set of limitations. Thus, detection methods that combine non-invasive detection, label-free detection, high sensitivity and high specificity are urgently needed to aid clinical diagnosis. Optical diagnostics and imaging are increasingly being employed in healthcare settings in a variety of sectors. Raman scattering can assess changes in molecular signatures in cancer cells or tissues based on the interaction with vibrational modes of common molecular bonds. Due to the advantages of label-free, strong chemical selectivity, and high sensitivity, Raman scattering, especially coherent Raman scattering microscopy imaging with high spatial resolution, has been widely used in biomedical research. And quantity studies have shown that it has a good application in the detection and diagnosis of bladder can-cer, renal clear cell carcinoma, prostate cancer, and other cancers. In this paper, several nonlinear imaging techniques based on Raman scattering technology are briefly described, including Raman spectroscopy, coherent anti-Stokes Raman scattering, stimulated Raman scattering, and surface-enhanced Raman spectroscopy. And we will discuss the application of these techniques for detecting urologic malignancy. Future research directions are predicted using the advantages and limitations of the aforesaid methodologies in the research. For clinical practice, Raman scattering technology is intended to enable more accurate, rapid, and non-invasive in early diagnosis, intraoperative margins, and pathological grading basis for clinical practice.
Humans ; Male ; Microscopy/methods* ; Radiopharmaceuticals ; Spectrum Analysis, Raman/methods* ; Technology ; Urologic Neoplasms/diagnosis*

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*

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*

Recording the highly diverse and dynamic activities in large populations of neurons in behaving animals is crucial for a better understanding of how the brain works. To meet this challenge, extensive efforts have been devoted to developing functional fluorescent indicators and optical imaging techniques to optically monitor neural activity. Indeed, optical imaging potentially has extremely high throughput due to its non-invasive access to large brain regions and capability to sample neurons at high density, but the readout speed, such as the scanning speed in two-photon scanning microscopy, is often limited by various practical considerations. Among different imaging methods, light field microscopy features a highly parallelized 3D fluorescence imaging scheme and therefore promises a novel and faster strategy for functional imaging of neural activity. Here, we briefly review the working principles of various types of light field microscopes and their recent developments and applications in neuroscience studies. We also discuss strategies and considerations of optimizing light field microscopy for different experimental purposes, with illustrative examples in imaging zebrafish and mouse brains.
Animals ; Mice ; Microscopy/methods* ; Zebrafish ; Neurons/physiology* ; Brain/physiology* ; Neurosciences

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.
Agrobacterium ; Blotting, Southern ; Digestion ; Genome ; Genomics ; Methods ; Microscopy, Fluorescence ; Oxidoreductases ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Wounds and Injuries