Autofluorescence has great advantage on detecting premalignant lesions and early cancers which are not detectable by conventional white light endoscopy (WLE). In this review, the recent advances in autofluorescence for diagnosis of precancerous lesions and early cancers are presented. Varieties of endogenous fluorophores in biological tissues, the potential mechanisms of the autofluorescence differences between normal and abnormal tissues, the selection of light source and optimal excitation wavelengths, and effective algorithms for processing autofluorescence data are highlighted. Finally, the shortages and improvement directions of autofluorescence technique for the diagnosis of precancerous lesions and early cancers are briefly discussed.
ObjectiveTo evaluate the value of real-time indocyanine green fluorescence imaging navigation (ICG-FIN) in laparoscopic rectal cancer surgery. MethodsThe patients who adopted ICG-FIN during laparoscopic rectal cancer surgery in the Department of Anorectal Surgery of Xuzhou Central Hospital from April 2022 to June 2023 according to the inclusion and exclusion criteria (ICG-FIN group) were collected, meanwhile matching (1∶1) of patients who did not adopt ICG-FIN during laparoscopic surgery from January 2021 to May 2022 (control group). The general data, surgical conditions, intraoperative and postoperative outcomes between the two groups were compared. ResultsThere were 62 patients in the ICG-FIN group and 62 patients in the control group. There were no statistical differences in the gender, age, body mass index, comorbidities, and so on between the two groups (P>0.05). The tumor localization, lymph node tracing, fluorescence imaging of the intended resection of intestinal tract and anastomotic site were observed in the ICG-FIN group. Seven patients (11.3%) had changed in the intended resection of intestinal anastomotic line during surgery, while there were no changes of the surgical plan in the control group. There were no statistical differences (P>0.05) in terms of surgical method, operative time, intraoperative bleeding, proportion of ileostomy, time of the first postoperative exhaust, postoperative hospital stay, and incidence of short-term complications between the two groups. Compared with the control group, the incidence of anastomotic leakage was lower (P=0.012), and the number of lymph nodes cleaned was more (P=0.016) in the ICG-FIN group. However, there was no statistical difference in the number of positive lymph nodes detected between the two groups (P=0.343). ConclusionsAccording to the results of this study, ICG-FIN is a reliable and effective method during laparoscopic rectal cancer surgery, which can accurately localize tumor, trace and guide lymph node dissection. Real-time evaluation of intestinal blood flow perfusion is of great practical value in reducing anastomotic leakage.
ObjectiveTo summarize the application status and progress of indocyanine green fluorescence imaging in laparoscopic anatomic liver resection , and to analyze its advantages, limitations, and prospects.MethodThe literatures about indocyanine green fluorescence imaging in laparoscopic anatomic liver resection were reviewed.ResultsIndocyanine green fluorescence imaging had been preliminarily used in the operation of liver tumors and had shown its unique value in the anatomical liver resection, providing a new way to reduce the recurrence of liver cancer, improve the therapeutic effect, and prolong the survival time of patients.ConclusionsThe clinical application of indocyanine green fluorescence imaging in anatomic liver resection is still at the stage of development and popularization. Although it has unique advantages and development potential, it needs to be further improved in the aspects of tissue penetration, specificity, and staining success rate.
Objective To insure early detection and hence efficient prevention of allograft rejection in transplanted heart, investigate possible applications of NAD(P)H fluorescence components analysis at the level of living cardiac cells to propose new approaches for diagnosis of rejection. Methods NAD(P)H was studied for noninvasive fluorescent probing of the mitochondrial function. Human cardiomyocyte were isolated from one additional endomyocardial biopsy (EMB) of 14 pediatric patients with heart ransplantation. Rat cardiomyocyte (n=5, 13-14 week old) were also isolated by the same approach for human myocytes. Autofluorescence(AF) was recorded in living cardiomyocytes following excitation with 375 nm UVlight and detection by spectrallyresolved time correlated single photon counting (TCSPC), based on the simultaneous measurement of the fluorescence spectra and lifetimes. Rat cardiac cells were divided into four groups: normoxic condition, normoxia with Rotenone, ischemic condition and ischemia with Rotenone. Comparison of cardiomyocyte AF between human and rat; compared kinetics of rat cardiomyocytes AF in normoxic conditions to ischemiamimicking ones, induced at physiological temperatures by reducing cell pH and oxygen content; comparison of cardiomyocyte AF dynamic changes in transplanted pediatric patients presenting either no rejection (R0) or mild rejection (R1). Results We have achieved appropriate isolation of living cardiomyocytes from human biopsies, as well as from rat cardiac tissues and determined their AF. At least a 3-exponential decay with 0.5-0.7ns, 1.9-2.4 ns and 9.0-15.0 ns lifetime pools is necessary to describe human cardiomyocyte AF within 420560 nm spectral range. Rat cardiomyocyte steadystate AF in ischemiamimicking condition was significantly increased when compared normoxic ones (Plt;0.05); application of Rotenone induced a significant increase in AF intensity in ischemic and normoxic condition, however no significant difference between the two groups (Plt;0.05).Human cardiomyocyte AF was found significantly lower in comparison to experimental rat model in the same condition(Plt;0.05). A correlation between changes in steadystate NAD(P)H fluorescence and rejection grades was found when comparison of R1 to R0. R1 showed significantly increased fluorescence intensity (Plt;0.05), without change in the spectra shape, results can be comparable to the effect of ischemiamimic conditions. Conclusion Our studies clearly demonstrated that spectrallyresolved fluorescence spectral analysis coupled to fluorescence lifetime are high sensitive approaches to examine mitochondrial metabolic oxidative state directly in living human cardiomyocytes with good reproducibility. Human cardiomyocytes are more metabolically active than the rat ones, while this activity (and thus ATP production) seems lowered during rejection process. In perspective, the advantage of this method is the possibility of its combination to multiphoton confocal microscopy, which can result in the adaptation of this approach directly to tissue biopsy, as well as in vivo directly via cardiac catheterization without the necessity of cell isolation. This approach provides promising new tool for clinical diagnosis and treatment of allograft rejection, and will enhance our knowledge about cardiomyocyte oxidative metabolism and/or its dysfunction at a cellular level.
Objective To analyze the variation of intestinal microflora in patients with colorectal cancer by SYBR GreenⅠreal-time fluorescence quantitative PCR and reveal the role and significance of intestinal microflora in the colorectal cancer-associated molecular pathogenesis. Methods A set of 16S rRNA gene group of species-specific primers for Bifidobacterium spp., Lactobacillus group, Escherichia coli, and ddl gene-targeted species-specific primers for Enterococcus faecalis and feces Enterococcus were designed. Patients with colorectal cancer (colorectal cancer group, n=30) and healthy volunteers (normal control group, n=30) were included and whose feces were collected to extract bacterial genome DNA. SYBR GreenⅠ real-time fluorescence quantitative PCR was used to analyze the five mentioned bacterial amounts. Results Level of Bifidobacterium spp. (4.52±0.49) and Lactobacillus group (5.46±0.12) in colorectal cancer group were significantly lower than those （9.25±0.83 and 7.45±0.37） of normal control group (Plt;0.05), whereas levels of Escherichia coli (5.82±0.47), Enterococcus faecalis (10.6±0.30) and feces Enterococcus (5.74±0.16) in colorectal cancer group were significantly higher than those （4.68±0.32, 4.95±0.24, and 5.03±0.43） of normal control group (Plt;0.05). Conclusions The fecal microflora composition of patients with colorectal cancer is significantly decreased in Bifidobacterium spp. and Lactobacillus group, whereas increased in Escherichia coli, Enterococcus faecalis, and feces Enterococcus. These data underline that the occurrence and progress of colorectal cancer may be related to intestinal microflora.
ObjectiveTo explore the clinical application of near-infrared autofluorescence (NIRAF) detection technology in protecting the parathyroid glands and the research progress on the autofluorescent substances. MethodThe recent literature on clinical application of NIRAF detection technology in protecting the parathyroid glands and the identification of fluorescent substances, both domestically and internationally, was conducted. ResultsThe majority of current studies indicate that NIRAF detection technology can effectively assist surgeons in identifying parathyroid tissue, improve the accuracy of intraoperative parathyroid identification, and reduce postoperative complications such as hypocalcemia. However, a small number of studies have found that the use of NIRAF detection technology during surgery does not significantly reduce postoperative complications in thyroid surgery patients, especially in those with secondary hyperparathyroidism. Current research on autofluorescent substances in the parathyroid glands remains relatively limited, with proteins such as the calcium-sensing receptor and vitamin D receptor being considered potential sources of fluorescence emitted by the parathyroid glands under near-infrared light excitation. ConclusionsBased on the reviewed literature, NIRAF detection technology for parathyroid gland identification has demonstrated significant effectiveness in intraoperative identification of parathyroid tissue and reduction of postoperative complications. However, limitations such as insufficient accuracy in patients with hyperparathyroidism and lack of user-friendliness restrict its clinical application. Therefore, future research should focus on identifying the endogenous fluorescent substances in the parathyroid glands and their luminescence mechanisms. This will enable targeted improvements in fluorescence detection technology, further enhancing the accuracy and convenience of intraoperative parathyroid detection, ultimately benefiting patients more significantly.
Objective To explore the accuracy and efficiency of indocyanine green fluorescence (ICGF) imaging in evaluating blood perfusion of parathyroid gland (PG) during total thyroidectomy. Methods Seventy patients who underwent total thyroidectomy and bilateral central lymph node dissection for papillary thyroid carcinoma (PTC) from March 2021 to December 2021 were enrolled and randomly divided into experimental group (ICGF imaging, n=35) and control group (normal treatment, n=35). Blood perfusion of PGs was evaluated by ICGF imaging and naked eye in each group respectively. The perfusion of PGs, incidence of hypoparathyroidism, and number of autotransplanted PGs were analyzed between the two groups. Results There was no difference between two groups in the incidence of transient hypoparathyroidism (P=0.339), and no one occurred permanent hypoparathyroidism. More PGs were autotransplanted in the experimental group compared to the control group (P＜0.001). At least one PG with good perfusion in the experimental group predicted an extremely high rate of normal parathyroid hormone levels of the patients postoperatively than the control group (P=0.003). Conclusion ICGF imaging can evaluate the blood perfusion of PGs accurately and guide their autotransplantation.
ObjectiveTo observe the detection of circulating tumor cells (CTCs) in peripheral blood of patients with gastric cancer, and to investigate the relationship between the CTCs and clinicalpathological features of gastric cancer. MethodsSixty cases of gastric cancer from September 2011 to September 2013 of our hospital were as the research object, and compared with 40 cases of benign gastric disease over the same period. These patients' venous blood were collected, the CTCs counts were determined by using the CellTracks AutoPrep fluorescence scaning system, and the relationship between CTCs and clinicopathological features of gastric cancer was analyzed. ResultsThe detection rate of CTCs in gastric cancer patients was 70.0% (42/60), in control group was 7.5% (3/40). The positive rate of CTCs in peripheral blood of patients with gastric cancer was significantly higher than that of benign gastric disease (P<0.05). The positive rate of CTCs in peripheral blood were no correlated with gender, age, N staging, distant metastasis, tumor size, and vascular invasion (P>0.05), but were correlated with the TNM staging of tumor and degree of differentiation (P<0.05). The cumulative survival rates of 12 months and 18 months after operation in CTCs negtive patients with gastric cancer were higher than that CTCs positive patients (P<0.05). ConclusionsThe detection of CTCs is easy to manage and repeatable. The positive rate of CTCs in gastric patients is higher, which can reflect the progression of tumor and serve as the prognostic index in gastric cance patients.
ObjectiveTo evaluate efficacy and safety of laparoscopic Glissonean pedicle transection anatomic hepatectomy using indocyanine green (ICG) fluorescence imaging.MethodThe retrospective analysis was made on the preoperative clinical data, surgical treatment and postoperative status of a patient with hepatocellular carcinoma who underwent the laparoscopic Glissonean pedicle transection anatomic hepatectomy using the ICG fluorescence imaging technology in the Department of Liver Surgery, West China Hospital of Sichuan University.ResultsAccording to the preoperative history, imaging and laboratory examinations, the diagnosis of hepatocellular carcinoma was considered. The intraoperative exploration revealed that there was only one tumor located in the segment Ⅳ and was superficial. The ICG fluorescence imaging was used to perform the Glissonean pedicle transection anatomic hepatectomy. The postoperative pathological diagnosis was consistent with hepatocellular carcinoma without serious complications. The patient recovered well. No recurrence was found in the follow-up period up to 6 months.ConclusionsLaparoscopic Glissonean pedicle transection anatomic hepatectomy using ICG fluorescence technology can be used as a safe and precise treatment to solve problems such as bleeding during operation, difficult determination of tumor boundary, and whether having residual tumor in surgical margin.
ObjectiveTo analyze the feasibility, advantages and disadvantages of the fluorescence method and the inflation-deflation method in defining the intersegmental plane during thoracoscopic lung segmental resection.MethodsFrom February to October 2018, 60 patients underwent thoracoscopic anatomical segmentectomy in Thoracic Surgery Department of Nanjing Chest Hospital, with 28 males and 32 females, aged from 25 to 82 years. Three-dimension computed tomography bronchography and angiography was used to reconstruct pulmonary vessels, bronchus and virtual intersegmental plane. Among them, 20 patients used the fluorescence method to define the intersegmental plane, and the other 40 patients used the traditional inflation-deflation method to define the intersegmental plane.ResultsFluorescent injection of indocyanine green (ICG) showed a clear intersegmental line with a duration sufficient to complete the label. With the fluorescence method, the intersegmental plane occurrence time was significantly shortened (10.75±3.78 s vs. 988.00±314.24 s, P<0.001) and had satisfactory repeatability. The lungs did not need to be inflated, which was convenient for the operation. And the operation time was shortened (108.75±31.28 min vs 138.00±32.47 min, P=0.002). No obvious ICG injection-related concurrency symptoms was found.ConclusionCompared with the traditional inflation-deflation method, the fluorescence method can display the intersegmental line quickly, accurately and clearly, reduce the difficulty of surgery, shorten the operation time, and provide reliable technical support for thoracoscopic anatomical segmentectomy. The fluorescence is a safe and effective method that is worthy of clinical application.