Abstract: Objective Using Amplex red fluorometric assay to detect the lysyl oxidase (LOX) enzyme activity in tissue engineered heart valve (TEHV). Methods Porcine aortic valves were decellularized with trypsin+ethylene diaminetetraacetic acid(EDTA), TritonX-100, and RNaseⅠ＋DNaseⅠ, then they were seeded by myo-fibroblasts that harvested from rats. Then they were fed with Dulbecco’s modified Eagle medium (DMEM) which contained high glucose for 27 days, they were fed with phenol red-free and serumfree DMEM for 24 hours, and the medium was harvested and used for LOX enzyme activity assays with the Amplex red fluorometric assay. And reverse transcription-polymerase chain reaction (RT-PCR) technique was used to analyze the expression of LOXmRNA in TEHV. Results All the samples produced measurable amounts of active LOX enzyme. The fluorescence units were 45.60±1.66, and the corresponding concentration of LOX enzyme were 0.123±0.003μg/ml. At the same time, all the samples expressed LOXmRNA. The expression of LOXmRNA was corresponding to the results of the Amplex red fluorometric assay. Conclusion It is feasible to detect the LOX enzyme activity in TEHV with the Amplex red fluorometric assay. And this assay gives a way to reflect that LOX plays an important role in collagen cross-linking of extracellar matrix in TEHV.
ObjectiveTo verify the consistency between artificial interpretation and automatic interpretation by HELIOS automatic immunofluorescence system by comparing their results on the same antinuclear antibodies (ANA) fluorescent slides, and analyze the application of automatic interpretation clinically. MethodA total of 281 ANA fluorescent slides of 281 impatients or outpatients in February 2015 were analyzed by HELIOS automatic immunofluorescence system and artificial interpretation respectively. As HELIOS could only determine the titer not the fluorescence type, only the negative or positive results qualitatively and the titer of ANA positive slides were analyzed. ResultsThere was no statistically significant difference between HELIOS automatic immunofluorescence system and artificial interpretation in negative or positive rate qualitatively (P>0.05) . The total coincidence rate was 98.9%, the positive coincidence rate was 99.5%, and the negative coincidence rate was 97.4%, and the kappa coefficient was 0.973. The difference of titer between the two groups had no statistical significance (P>0.05) . ConclusionsThe results of HELIOS automatic Immunofluorescence system and artificial interpretation are in good consistency. HELIOS automatic immunofluorescence system is suitable for clinical use as its high degree of automation, simple operation and result reliability.
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 To evaluate the effectiveness and safety of indocyanine green fluorescence method versus modified inflation-deflation method for thoracoscopic anatomic segmentectomy. Methods CNKI, Wanfang Database, China Biomedical Literature Database, Web of Science, Cochrane Library, EMbase, PubMed, Clinicaltrials.gov, were searched from 1 January 2000 to 1 May 2023, and controlled studies between indocyanine green fluorescence and modified inflation deflation method in thoracoscopic segmentectomy were collected. Meta-analysis was performed using Stata14MP and RevMan5.4. Results A total of 10 articles, including 1 156 patients, were identified. In thoracoscopic anatomic segmentectomy, indocyanine green fluorescence method had an advantage over modified inflation deflation method. The total incidence of postoperative complications decreased (OR=0.51, 95%CI 0.36 to 0.71, P<0.0001). The incidence of air leaks decreased (OR=0.50, 95%CI 0.31 to 0.80, P=0.004), the operation time shortened (MD=−25.81, 95%CI −29.78 to −21.84, P<0.00001), the length of postoperative hospital stays shortened (MD=−0.98, 95%CI −1.57 to −0.39, P=0.001), the rate of clear displaying for intersegmental boundary line increased (OR=5.79, 95%CI 2.76 to 12.15, P<0.00001). The difference was statistically significant. Conclusion Compared with modified inflation deflation method, indocyanine green fluorescence method can quickly and clearly display the intersegmental boundary line, reduce the difficulty of surgery, shorten the operation time, reduce the length of postoperative hospital stay, and provide reliably technical support for thoracoscopic anatomic segmentectomy. It is an effective and safe method, which is worthy of extensive application.