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*

OBJECTIVE: To assess the application of probe-based confocal laser endomicroscopy (pCLE) in diagnosis of gastric carcinoma and precancerous lesions. METHODS: Patients underwent pCLE in the First Affiliated Hospital of Zhejiang University School of Medicine during December 2013 and November 2014 and in the First Affiliated Hospital of Zhejiang Chinese Medical University during January 2014 and December 2017 were enrolled. The consistency between pCLE diagnosis and pathological diagnosis of gastric lesions, including atrophic gastritis, gastric intestinal metaplasia, low-grade intraepithelial neoplasia and high-grade intraepithelial neoplasia (including gastric carcinoma) was analyzed. RESULTS: Totally 154 gastric lesions from 119 patients were detected by pCLE. Using pathological diagnosis as gold standard, the sensitivity, specificity, coincidence rate and κ value of pCLE diagnosis for atrophic gastritis were 94.34%, 91.09%, 92.21%and 0.83; those indicators for gastric intestinal metaplasia were 84.47%, 92.16%, 87.01% and 0.72. The coincidence rate and κ value of pCLE diagnosis of complete gastric intestinal metaplasia were 0.75 and 0.49; for incomplete gastric intestinal metaplasia were 0.79 and 0.48, respectively. The sensitivity, specificity, coincidence rate and κ value of pCLE diagnosis for low-grade intraepithelial neoplasia were 85.29%, 87.50%, 87.01%and 0.66; those for high-grade intraepithelial neoplasia (including gastric carcinoma) were 95.83%, 97.17%, 96.75%and 0.92. CONCLUSIONS pCLE can be used for diagnosis of gastric carcinoma and pericancerous lesions and also for typing of gastric intestinal metaplasia.
Carcinoma ; diagnostic imaging ; Endoscopy, Gastrointestinal ; Humans ; Metaplasia ; Microscopy, Confocal ; Precancerous Conditions ; diagnostic imaging ; Sensitivity and Specificity ; Stomach ; diagnostic imaging ; pathology ; Stomach Neoplasms ; diagnostic imaging

BACKGROUND: Respiratory viral infections are the leading cause of asthma exacerbations. Eosinophil activation results in the formation of eosinophil extracellular traps (EETs), which release web-like structures of DNA and proteins that bind, disarm and extracellularly kill pathogens. OBJECTIVE: We investigated whether the respiratory syncytial virus (RSV) in vitro could induce EETs in bronchoalveolar lavage fluid eosinophils in a murine model of asthma. METHODS: BALB/cJ mice (6–8 weeks old) were sensitized with 2 subcutaneous injections of ovalbumin (20 μg) on days 0 and 7, followed by three intranasal challenges with ovalbumin (100 μg) on days 14, 15, and 16 of the protocol. The control group received Dulbecco's phosphate-buffered saline. Bronchoalveolar lavage fluid eosinophils of ovalbumin group or control group were stimulated with RSV (103 PFU/mL) in vitro for 3 hours. After that, culture supernatant was collected to perform the analyses proposed in this study. RESULTS: We verified an increase in extracellular DNA concentration in bronchoalveolar lavage fluid eosinophils from ovalbumin group stimulated with RSV (10³ PFU/mL) in vitro, which was confirmed by confocal microscopy. We demonstrated that most cells are negative for annexin V and propidium iodide in all groups evaluated. Also, RSV in vitro decreased interferon-ɣ in culture supernatant when compared to the ovalbumin group. CONCLUSION: In this study, we demonstrated for the first time that RSV in vitro induces EETs formation in eosinophils from asthmatic mice.
Animals ; Annexin A5 ; Asthma ; Bronchoalveolar Lavage Fluid ; DNA ; Eosinophil Peroxidase ; Eosinophils ; Extracellular Traps ; In Vitro Techniques ; Inflammation ; Injections, Subcutaneous ; Mice ; Microscopy, Confocal ; Ovalbumin ; Propidium ; Respiratory Syncytial Viruses

OBJECTIVE: This work aims to uncover the promoting effect of 17% ethylenediaminetetraacetic acid (EDTA) irrigation on the dentin adhesion of Enterococcus faecalis (E. faecalis). METHODS: Forty-eight half split samples and twelve dentin slices were randomly divided into three experimental groups and one control group. The experimental groups and the control group were soaked with EDTA in different time lengths and with normal saline, respectively. E. faecalis was inoculated, and its dentin adhesion was measured via scanning electron microscopy, confocal laser scanning microscopy (CLSM), colony forming unit counts, and histological Gram staining. RESULTS: According to histological Gram staining, the depth showed no statistically significant differences between 1 min group and the control group, 1 min group and 3 min groups (P>0.05). E. faecalis intruded in the dentine tubules (measured by CLSM), and the thickness of the biofilm on the dentin surface and the colony numbers of experimental groups were greater than those of the control group (P<0.05). The differences between the three experimental groups were statistically signi-ficant (P<0.05). CONCLUSIONS EDTA (17%) irrigation can promote E. faecalis adhesion to dentin. This adhesion would in turn prolong EDTA treatment time.
Biofilms ; Dentin ; Edetic Acid ; Enterococcus faecalis ; Microscopy, Confocal ; Root Canal Irrigants ; Sodium Hypochlorite

PURPOSE: The purpose of the present study was to compare the accuracy of four different metal copings fabricated by CAD/CAM technology and to evaluate clinical effectiveness. MATERIALS AND METHODS: Composite resin tooth of the maxillary central incisor was prepared for a metal ceramic crown and duplicated metal die was fabricated. Then scan the metal die for 12 times to obtain STL files using a confocal microscopy type oral scanner. Metal copings with a thickness of 0.5 mm and a cement space of 50 µm were designed on a CAD program. The Co-Cr metal copings were fabricated by the following four methods: Wax pattern milling & Casting (WM), Resin pattern 3D Printing & casting (RP), Milling & Sintering (MS), Selective laser melting (SLM). Silicone replica technique was used to measure marginal and internal discrepancies. The data was statistically analyzed with One-way analysis of variance and appropriate post hoc test (Scheffe test) (α=.05). RESULTS: Mean marginal discrepancy was significantly smaller in the Group WM (27.66 ± 9.85 µm) and Group MS (28.88 ± 10.13 µm) than in the Group RP (38.09 ± 11.14 µm). Mean cervical discrepancy was significantly smaller in the Group MS than in the Group RP. Mean axial discrepancy was significantly smaller in the Group WM and Group MS then in the Group RP and Group SLM. Mean incisal discrepancies was significantly smaller in the Group RP than in all other groups. CONCLUSION: The marginal and axial discrepancies of the Co-Cr coping fabricated by the Wax pattern milling and Milling/Sintering method were better than those of the other groups. The marginal, cervical and axial fit of Co-Cr copings in all groups are within a clinically acceptable range.
Ceramics ; Crowns ; Dental Marginal Adaptation ; Freezing ; Incisor ; Methods ; Microscopy, Confocal ; Printing, Three-Dimensional ; Replica Techniques ; Silicon ; Silicones ; Tooth ; Treatment Outcome

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