ABSTRACT
BACKGROUND AND PURPOSE: Cutaneous nerve biopsies based on two-dimensional analysis have been regarded as a creditable assessment tool for diagnosing peripheral neuropathies. However, advancements in methodological imaging are required for the analysis of intact structures of peripheral nerve fibers. A tissue-clearing and labeling technique facilitates three-dimensional imaging of internal structures in unsectioned, whole biological tissues without excessive time or labor costs. We sought to establish whether a tissue-clearing and labeling technique could be used for the diagnostic evaluation of peripheral neuropathies. METHODS: Five healthy individuals and four patients with small-fiber neuropathy (SFN) and postherpetic neuralgia (PHN) were prospectively enrolled. The conventional methods of indirect immunofluorescence (IF) and bright-field immunohistochemistry (IHC) were adopted in addition to the tissue-clearing and labeling method called active clarity technique-pressure related efficient and stable transfer of macromolecules into organs (ACT-PRESTO) to quantify the intraepidermal nerve-fiber density (IENFD). RESULTS: The mean IENFD values obtained by IF, bright-field IHC, and ACT-PRESTO in the healthy control group were 6.54, 6.44, and 90.19 fibers/mm², respectively; the corresponding values in the patients with SFN were 1.99, 2.32, and 48.12 fibers/mm², respectively, and 3.06, 2.87, and 47.21 fibers/mm², respectively, in the patients with PHN. CONCLUSIONS: This study has shown that a tissue-clearing method provided not only rapid and highly reproducible three-dimensional images of cutaneous nerve fibers but also yielded reliable quantitative IENFD data. Quantification of the IENFD using a tissue-clearing and labeling technique is a promising way to improve conventional cutaneous nerve biopsies.
Subject(s)
Humans , Biopsy , Fluorescent Antibody Technique, Indirect , Imaging, Three-Dimensional , Immunohistochemistry , Methods , Nerve Fibers , Neuralgia, Postherpetic , Peripheral Nerves , Peripheral Nervous System Diseases , Prospective StudiesABSTRACT
Objective@#Preliminary study on the clinical effect of preoperative ultrasound endoscopy combined with staining labeling technique to locate the actual boundary of esophageal and gastric cancer@*Methods@#From September 1, 2015 to October 30, 2017, 18 patients with esophageal adenocarcinoma were enrolled in this study. The actual boundaries of esophageal and gastric-derived adenocarcinoma lesions were localized by endoscopic ultrasonography and staining. There were 10 males and 8 females. After completing the preoperative examination, 1-2 days before operation, endoscopic ultrasonography was used to locate the edge of the lesion. Two point injection of carbon nano suspension was used to mark the location of 1cm at the longest distance from the longitudinal axis of the tumor. According to the length of longitudinal axial staining, the thoracotomy was performed. Intraoperative proximal margin resection was used to send frozen pathology. According to the results of freezing, the operation was decided. After the operation, the specimens from the margin of the tumor were segmented into paraffin section, which was about 0.5cm in each segment, and the tumor cells were observed under the electron microscope at all levels of the paraffin sections.@*Results@#The average time of preoperative endoscopic ultrasonography staining was(10.16±1.38) min, and the diameter of nano carbon diffusion was(1.43±0.41)cm. All patients in the operation could clearly see the nano carbon staining area under the naked eye. In the field, the average time of locating lesions was(1.27±0.53)min. 5 patients underwent thoracoabdominal surgery and 13 underwent abdominal surgery. The average length of the cut margin of the tumor was(4.74±1.12)cm, and the frozen pathology of the incision margin was negative, and no additional operation was performed. The routine pathology confirmed that all the specimens were negative.@*Conclusion@#The staining and labeling technique for adenocarcinoma of the esophagogastric junction under endoscopic ultrasonography can detect the tumor edge and the scope of invasion accurately. It provides guidance and guarantee for the smooth implementation of AEG precision surgery. It is a safe, rapid and effective positioning technique.
ABSTRACT
Objective Preliminary study on the clinical effect of preoperative ultrasound endoscopy combined with stai-ning labeling technique to locate the actual boundary of esophageal and gastric cancer Methods From September 1, 2015 to October 30, 2017, 18 patients with esophageal adenocarcinoma were enrolled in this study. The actual boundaries of esophage-al and gastric-derived adenocarcinoma lesions were localized by endoscopic ultrasonography and staining. There were 10 males and 8 females. After completing the preoperative examination, 1-2 days before operation, endoscopic ultrasonography was used to locate the edge of the lesion. Two point injection of carbon nano suspension was used to mark the location of 1cm at the lon-gest distance from the longitudinal axis of the tumor. According to the length of longitudinal axial staining, the thoracotomy was performed. Intraoperative proximal margin resection was used to send frozen pathology. According to the results of freezing, the operation was decided. After the operation, the specimens from the margin of the tumor were segmented into paraffin section, which was about 0. 5cm in each segment, and the tumor cells were observed under the electron microscope at all levels of the paraffin sections. Results The average time of preoperative endoscopic ultrasonography staining was(10. 16 ± 1. 38) min, and the diameter of nano carbon diffusion was(1.43 ±0.41)cm. All patients in the operation could clearly see the nano carbon staining area under the naked eye. In the field, the average time of locating lesions was(1.27 ±0.53)min. 5 patients under-went thoracoabdominal surgery and 13 underwent abdominal surgery. The average length of the cut margin of the tumor was(4. 74 ±1.12)cm, and the frozen pathology of the incision margin was negative, and no additional operation was performed. The routine pathology confirmed that all the specimens were negative. Conclusion The staining and labeling technique for adeno-carcinoma of the esophagogastric junction under endoscopic ultrasonography can detect the tumor edge and the scope of invasion accurately. It provides guidance and guarantee for the smooth implementation of AEG precision surgery. It is a safe, rapid and effective positioning technique.
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Objective To investigate the sensitivities and specificities of enzyme-linked inununosorbent assay(ELISA)and immunogold labeling technique in the detection of anti-human immunodeficiency virus antibody.Methods(1)Sensitivity test:15 sera from patients with acquired immunodeficiency syndrome(AIDS)diagnosed by western blotting were diluted proportionately,we detected the anti-human immunodeficiency virus antibody in all diluted sera using ELISA and immunogold labeling technique,and recorded the maximum dilutions of the two methods that the anti-human immunodeficiency virus antibody was positive.(2)Specificity test:200 sera from patients with nonAIDS were detected by ELISA and immunogold labeling technique,and we recorded the negative rates of the two methods.Results(1)Sensitivity test:The maximum dilution of ELISA was significantly higher than that of immunogold labeling technique(P ＜ 0.05).(2)Specificity test:The negative rates of the two methods were 100％,and there was no significant difference between the two methods.Conclusion The sensitivity of ELISA in the detection of antihuman immunodeficiency virus antibody is better than that of immunogold labeling technique,but the sensitivities and specificities of the two methods are consistent when the serum is detected,so the immunogold labeling technique is able to replace ELISA as a screening test for AIDS.