Supported lipid bilayers (SLBs) have been widely used in biomedical and bioengineering research in vitro because its structure and function are similar to natural cell membrane. A fluorescence recovery after photobleaching (FRAP) technique was used to measure the lateral diffusion of the SLBs composed of 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1, 2-dioleoyl-sn-glycero-3-［(N-(5-amino-1-carboxyp-entyl) iminodiacetic acid)］ (DGS-NTA) on the glass slide, and the effects of the DOPC-to-DGS-NTA ratio, small unilamellar vesicles (SUV) producing method, sizes of bleaching areas and concentrations of loading proteins on the SLBs fluidity and diffusion coefficient were studied systematically in this paper. The results demonstrated that: (1) SUV made by probe sonication exhibited more uniform and smaller size compared with that made by film extrusion, but the whole process of SLBs formation must not be exposed to air. (2) The fluorescence recovery rate and diffusion coefficient of the SLBs decreased with the increasing bleaching area size. With the mole ratio of DOPC to DGS-NTA decreasing from 98∶2 to 84∶16, the fluidity and fluorescence recovery degree decreased gradually, and the SLBs would lose its fluidity if the ratio reached to 82∶18. (3) The average fluorescence intensity of SLBs increased linearly with the loading protein concentration (10–40 nmol·L–1), and the protein showed good mobility on the SLBs. The study would provide a good platform of bio-membrane for further research on interactions among cell membrane molecules and subsequent signals response.
ObjectiveTo summarize the research progress of near infra-red fluorescence imaging (NIRFI) in biliary tract surgery, and to provide protection for improvements of therapeutic effect and safety of biliary tract surgery.MethodThe relevant literatures about studies on NIRFI in the biliary tract surgery in recent years were reviewed.ResultsThe NIRFI had been preliminarily used in the surgical treatment of benign and malignant biliary diseases, and had shown its unique value in cholangiography. It provided a new method for effectively avoiding surgical complications, shortening operation time, reducing the rate of conversion to open surgery, evaluating blood supply of bile duct and improving the safety of operation.ConclusionsNIRFI has achieved notable successes in treatment of biliary tract diseases. With future application of fluorescence imaging in near infra-red Ⅱ window and new specific fluorescence targeting molecules, this technique will highlight its more important values in biliary surgery.
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.
Objective Using chemically extracted acellular methods to treat extracranial section of the canine whole facial nerve, to evaluated its effects on nerve structure and the removal extent of Schwann cells and myel in. Methods Twenty whole facial nerves were exposed from 10 canines [weighing (18 ± 3) kg]. The extracranial trunk of canine facial nerve and its branches (temporal branch, zygomatic branch, buccal branch, marginal mandibular branch, and cervical branch) were dissected under l ight microscope. Twenty facial nerves were divided into the experimental group (n=12) and control group (n=8) randomly. In experimental group, the nerve was extracted with the 3%TritonX-100 and 4% sodium deoxycholate. In control group, the nerve was not extracted. HE staining and immunofluorescence histological stainings for Hoechst33258, P75, Zero, and Laminin were performed. Results After histological staining, it was found that myel in and Schwann cells were removed from the facial nerve while the basal lamina tube remained intact. The whole canine facial nerves (one nerve trunk and multiple nerve branches) had the similar result. Conclusion The canine whole facial nerve has natural structure (one nerve trunk and multiple nerve branches) by extracted with chemically extracted acellular methods, so it is an available graft for repairing the defect of the whole facial nerve.
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.
Objective To observe preserving effect on myocytes in porcine aortic valve replacement with minimal extracorporeal circulation (MECC). Methods 7 pigs were collected as experimental animals and undertook aortic valve replacement with MECC. Morphological and immunofluorescence intensity changes of right atrial and left ventricular tissues were observed. Results HE staining showed that there were not significant changes and edema or injury of myocytes of right atriums and left ventricles between preoperation and postoperation. Immunofluorescence staining showed complement C3b/c in right atrial myocardial tissues after the operation were a little ber, and innate antibody IgG were a little ber in left ventricular myocardial tissues but similarly weak in right atrial myocardial tissues pre- and post-operation. There was not significant changes in HSPG staining in pre-and post-operative right atrial myocardial tissues, but HSPG were obviously weaker in left ventricular myocardial tissues after the operation. Conclusion MECC is effective on support of porcine aorta valve replacement.
ObjectiveTo investigate the application value of indocyanine green (ICG) fluorescence imaging technology for determining the blood supply of parathyroid in thyroid surgery.MethodsThe patients who underwent total thyroidectomy and bilateral central lymph node dissection for papillary thyroid carcinoma (PTC) from June 1, 2017 to January 1, 2018 were prospectively enrolled and then divided into a study group and control group randomly. The study group used the ICG fluorescence imaging technology to evaluate the blood supply of the parathyroid glands, while the control group assessed the blood supply by naked eyes, then determined that whether the parathyroid glands were retained in situ or autotransplanted. The incidence of hypoparathyroidism, length of hospital stay, and parathyroid hormone (PTH) were compared between the two groups.Results① A total of 60 patients with PTC were included in the study, and 30 patients in each group. There were no significant differences in the baseline informations of the two groups such as the gender, age, comorbidities, and preoperative PTH, Ca2+ levels, etc. (P>0.05). ② The ICG score of type A parathyroid glands (except type A3) was lower than that of type B parathyroid glands (0.99±0.38 versus 1.45±0.58, t=–2.395, P<0.05). ③ The length of postoperative hospital stay was shorter in the study group than in the control group (t=–2.159, P=0.035). ④ The ICG fluorescence imaging could significantly reduce the incidence of temporary hypoparathyroidism (χ2=5.079, P=0.024). The incidence of permanent hypoparathyroidism was not statistically different between the two groups (χ2=1.000, P=0.317), and only 1 case appeared in the control group. ⑤ There were no statistically significant differences in the PTH and serum Ca2+ levels at day 1, month 1, month 3, and month 6 after the surgery between the two groups (P>0.05). ConclusionICG fluorescence imaging technology could be used to determine blood supply of parathyroid in situ in real time during operation. Further studies are needed to confirm this conclusion.
Objective To explore the feasibility and accuracy of using indocyanine green fluorescence (ICGF) to identify the intersegmental plane after ligation of the target pulmonary vein during thoracoscopic segmentectomy. Methods From December 2022 to June 2023, the patients with pulmonary nodules undergoing video-assisted thoracoscopic anatomical segmentectomy with intersegmental plane displayed using ICGF after ligation of the target pulmonary vein by the same medical team in our hospital were collected. Preoperative three-dimensional reconstruction was used to identify the target segment where the pulmonary nodule was located and the anatomical structure of the arteries, veins, and bronchi in the target segment. The intersegmental plane was first determined by the inflation-deflation method after the target pulmonary vein was ligated during the operation. During the waiting period, the target artery and bronchus could be separated but not cut off. The inflation-deflation boundary was marked by electrocoagulation, and then ICGF was injected via peripheral vein to identify the intersegmental plane again, and the consistency of the two intersegmental planes was finally evaluated. Results Finally 32 patients were collected, including 14 males and 18 females, with an average age of 58.69±11.84 years, ranging from 25 to 76 years. The intersegmental plane determined by inflation-deflation method was basically consistent with ICGF method in all patients. All the 32 patients successfully completed uniportal thoracoscopic segmentectomy without ICGF-related complications or perioperative death. The average operation time was 98.59±20.72 min, the average intraoperative blood loss was 45.31±35.65 mL, and the average postoperative chest tube duration was 3.50±1.16 days. The average postoperative hospital stay was 4.66±1.29 days, and the average tumor margin width was 26.96±5.86 mm. Conclusion The ICGF can safely and accurately identify the intersegmental plane by target pulmonary venous preferential ligation in thoracoscopic segmentectomy, which is a useful exploration and important supplement to the simplified thoracoscopic anatomical segmentectomy.
This consensus aims to elucidate the applications of 3D fluorescence technology in thoracic surgery, summarizing its clinical value and prospects in areas such as sub-lobar resection, mediastinal structure protection, and lymph node dissection, providing a reference for the clinical practice of 3D fluorescence technology in thoracic surgery. Furthermore, this consensus is committed to advancing the research and innovation of domestically produced 3D fluorescence medical endoscopes. It seeks to clarify clinical practice needs and technical development directions, promote the widespread application of high-quality domestic medical endoscopes, and support the independent innovation and development of China's medical equipment industry.
ObjectiveTo evaluate diagnostic performance of crithidia luciliae immunofluorescence test (CLIFT), enzyme linked immunosorbent assay (ELISA), linear immunoassay (LIA) and chemiluminescence immunoassay (CLIA) for detection of anti-dsDNA antibodies for systemic lupus erythematosus (SLE).
MethodsA total of 178 sera[SLE (n=86), other systemic rheumatic diseases (n=62), and healthy individual (n=30)], from whom received treatment from July 2012 to June 2013, were tested by 4 different assay kits.
ResultsThe diagnostic performances of four methods for detecting anti-dsDNA antibodies for SLE were ELISA, CLIA, CLIFT and LIA, from higher to lower; while ELISA had the highest sensitivity (67.4%), and CLIA had the highest specificity (95.6%). The three test methods (ELISA, LIA, CLIA) had almost perfect concordance with the comparison method (CLIFT, Kappa >0.8). With cut-off values set at 95% of specificity, there was no statistical difference of sensitivity between ELISA and CLIA (58.1%, 60.5%; P>0.05).
ConclusionFour assays can be used for the clinical detection of anti-dsDNA antibodies, and the results have an almost perfect concordance. Different assays show various performances depending on the methods and cut-off values used.
