The role of near‐infrared fluorescence imaging with indocyanine green dye in pedicle division with the paramedian forehead flap

The paramedian forehead flap is considered the gold standard for nasal reconstruction following oncologic surgery. During the 21‐day delay in two‐stage surgery protocols, many patients report considerably reduced quality of life because of the pedicle. This prospective case series study examined the usefulness of near‐infrared (NIR) fluorescence with indocyanine green (ICG) for flap perfusion assessment and identified variables associated with time to flap perfusion. Ten patients (mean age 75.3 ± 11.6 years) with diagnosis of basal cell carcinoma (n = 9) or squamous cell carcinoma (n = 1) underwent intravenous indocyanine injection and NIR fluorescence imaging for assessment of flap vascularisation 2 to 3 weeks after stage 1 surgery. NIR fluorescence imaging showed 90% to 100% perfusion areas in all patients after 14 to 21 days. Early pedicle division occurred in two patients on postoperative days 14 and 16. One minor complication (wound healing disorder) was seen following flap takedown after 14 days. There were no associations between time to flap perfusion and defect size or flap area. NIR fluorescence imaging with ICG dye is a useful method for non‐invasive perfusion assessment when used in conjunction with clinical assessment criteria. However, a decision for early pedicle division may raise risk of complications in specific patient groups and must therefore be made with great care.     

Indocyanine green fluorescence imaging for laparoscopic complex upper urinary tract reconstructions: a comparative study

BackgroundTo describe our technique for using an intraureteral injection of indocyanine green (ICG) and visualization under near-infrared fluorescence (NIRF) to facilitate challenging upper urinary tract reconstructions (UUTRs) and to present the comparative outcomes.MethodsWe collected 36 patients who underwent laparoscopic UUTRs between April 2019 and March 2020, and we divided the patients into two groups based on the use of ICG (ICG group and non-ICG group). Demographic characteristics, perioperative outcomes, and functional outcomes were compared between the two groups.ResultsThere were 18 cases in the ICG group and 18 cases in the non-ICG group, respectively. There were no differences in the baseline characteristics between the two groups. The intraoperative time to identification of the ureter (TIU; 20.9±11.7 vs. 30.0±14.6 min, P=0.03) and length of postoperative hospital stay (LPHS; 11.1±3.0 vs. 16.6±10.0 days, P=0.03) were significantly shorter in the ICG group. There was also a trend for lesser time for locating the stricture (43.0±27.9 vs. 55.4±18.6 min, P=0.14) and lower estimated blood loss (EBL) in the ICG group patients (88.3±75.4 vs. 91.7±46.2 mL, P=0.22). During the mean 3.8-month follow-up for the ICG group and the 6.2-month for the non-ICG group, there was a trend for more severe complications in the non-ICG group.ConclusionsVisualizing intraureteral ICG under NIRF is useful in challenging UUTRs, allows for rapid ureteral identification and accurate real-time delineation of the ureteral stricture margins, and provides encouraging follow-up outcomes compared with those in the non-ICG group.     

Characterization of EGFP-labeled mesenchymal stem cells and redistribution of allogeneic cells after subcutaneous implantation

INTRODUCTION: Bone marrow mesenchymal stem cells (MSCs) are ideal target cells for cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by different methods. It is important to study the short-term fate of labeled allogeneic MSCs after subcutaneous implantation in rabbits in order to provide insights into the application of allogeneic MSCs for tissue regeneration. MATERIALS AND METHODS: Mesenchymal stem cells were labeled by two different methods in vitro and then were incubated with gelatin sponge. Autologous MSCs-Gelatin constructs and allogeneic MSCs-Gelatin constructs were subcutaneously implanted into 32 rabbits. The constructs were analyzed for the survival and migration of labeled MSCs at day 3, week 1, 3, and 5 post-implantation. RESULTS: EGFP was successfully expressed following transfection of MSCs with the retroviral vector pLEGFP-N1. In addition, EGFP-MSCs can be functionally induced into osteocytes, chondrocytes, and adipocytes in conditioned media. By weeks 3 after implantation, the labeled cells distributed extensively on the surface of gelatin sponge and gradually integrated into host tissues. EGFP-labeled MSCs were observed under fluorescence microscopy and BrdU-expressing cells were detected with immunohistochemical stain in allogeneic or autologous MSCs-Gelatin constructs during the initial five weeks after implantation. CONCLUSIONS: It is a simple and reliable way to trace the changes of MSCs in vivo by EGFP in cell transplantation and gene therapy. Allogeneic rabbit MSCs can survive for at least 5 weeks after subcutaneous implantation and maintain a strong ability of migration, indicating that allogeneic MSCs are of certain value in clinical application for temporary replacement.     

硬脊膜内背侧动静脉瘘的显微外科治疗

目的 探讨硬脊膜内背侧动静脉瘘的显微外科治疗及其疗效,初步探索术中吲哚菁绿荧光血管造影的应用.方法 2004年1月至2008年8月共有24例硬脊膜内背侧动静脉瘘患者进行了半椎板切除显微外科治疗并获得随访.其中2例为先接受介入栓塞治疗,后因复发再接受外科手术治疗.3例接受术中吲哚菁绿荧光血管造影.脊髓功能评价采用改良阿米诺夫量表,结合影像学检查结果对术后疗效进行综合评价.结果 术后平均随访36个月,影像学随访未见病变残留或复发.脊髓功能评价16例为优,阿米诺夫评分平均0.7分;4例为良,平均4.8分;3例为中,平均6.7分;1例为差,平均9.0分.综合疗效评价,治愈16例,改善6例,无变化2例.结论 单侧半椎板切除入路显微外科治疗足对硬脊膜内背侧动静脉瘘的有效治疗方法,术中吲哚菁绿荧光血管造影有助于手术.     

与基底细胞癌细胞凋亡相关的分子生物学研究--促凋亡分子Bad的克隆及其pEGFP-C3真核表达载体的构建

目的:克隆Bcl-2相关凋亡基因Bad并构建其真核表达载体,探讨其对肿瘤细胞凋亡的诱导作用。方法:采用RT-PCR的方法,扩增Bad基因全长片段。通过基因定向克隆,构建Bad基因的真核表达质粒载体。结果:经酶切鉴定、PCR及DNA序列测定鉴定,Bad基因表达质粒pEGFP-C3载体成功构建。结论:成功克隆Bcl-2相关促凋亡基因Bad并构建其真核表达载体pEGFP-C3-Bad,为进一步研究Bad在人肿瘤细胞系中的促凋亡作用奠定了实验基础。     

Cavernous antioxidant effect of green tea,epigallocatechin‐3‐gallate with/without sildenafil citrate intake in aged diabetic rats

This study aimed to assess the cavernous antioxidant effect of green tea (GT), epigallocatechin‐3‐gallate (EGCG) with/without sildenafil citrate intake in aged diabetic rats. One hundred and four aged male white albino rat were divided into controls that received ordinary chow, streptozotocin (STZ)‐induced aged diabetic rats, STZ‐induced diabetic rats on infused green tea, induced diabetic rats on epigallocatechin‐3‐gallate and STZ‐induced diabetic rats on sildenafil citrate added to EGCG. After 8 weeks, dissected cavernous tissues were assessed for gene expression of eNOS, cavernous malondialdehyde (MDA), glutathione peroxidase (GPx), cyclic guanosine monophosphate (cGMP), and serum testosterone (T). STZ‐induced diabetic rats on GT demonstrated significant increase in cavernous eNOS, cGMP, GPx and significant decrease in cavernous MDA compared with diabetic rats. Diabetic rats on EGCG demonstrated significant increase in cavernous eNOS, cGMP, GPx and significant decrease in cavernous MDA compared with diabetic rats or diabetic rats on GT. Diabetic rats on EGCG added to sildenafil showed significant increase in cavernous eNOS, cGMP and significant decrease in cavernous MDA compared with other groups. Serum T demonstrated nonsignificant difference between the investigated groups. It is concluded that GT and EGCG have significant cavernous antioxidant effects that are increased if sildenafil is added.     

携带报告基因EGFP的CXCR4 RNA干扰质粒的构建及生物活性的鉴定

目的构建携带报告基因EGFP的CXCR4RNA干扰质粒,证实其高效抑制靶基因表达的生物活性。方法设计有发夹状结构的两条DNA序列,用PCR扩增CXCR4特异性小干扰RNA(smallinterferingRNA,siRNA),转入带有增强型绿色荧光蛋白(EGFP)和启动子U6的pGensil-1质粒,转化DH5a菌株,提取质粒行酶切及测序鉴定。重组质粒转染PC-3m细胞株,检测绿色荧光蛋白表达及对细胞CXCR4表达的抑制情况。结果成功构建了CXCR4靶向的RNA干扰质粒pGensil-1/siCXCR4,在靶细胞可同时表达siRNA及绿色荧光蛋白。结论pGensil-1/siCXCR4质粒的成功构建对利用基因干扰技术治疗前列腺癌转移方面的研究奠定了一定的实验基础。     

Indocyanine green guided laser photocoagulation in patients with occult choroidal neovascularisation

AIMS: To determine whether indocyanine green (ICG) guided laser photocoagulation of occult choroidal neovascularisations (OCNV) is beneficial for patients with occult choroidal neovascularisation secondary to age related macular degeneration (AMD). METHODS: A prospective pilot study was performed in 21 eyes with OCNV secondary to AMD that could be identified extrafoveolarly or juxtafoveolarly in an early ICG angiographic study. Laser photocoagulation was applied to the neovascular membrane identified in the early ICG angiographic study. RESULTS: Visual acuity ranged from 20/400 to 20/20 (logMAR 0.54 (SD 0.29) before and hand movements and 20/30 (logMAR 0.81 (0.69)) at the last follow up after laser photocoagulation. During the follow up (30 (13) months) vision improved in four eyes (two lines), in seven eyes the initial visual acuity could be stabilised (two lines), in five eyes vision dropped moderately (three to five lines), and in five eyes vision decreased severely (six or more lines). Recurrences (seven patients) or persistent CNV (six patients) was observed in 13 patients. CONCLUSION: This preliminary study of ICG guided laser photocoagulation of occult extrafoveal and juxtafoveal choroidal neovascularisations suggests that this technique may improve the visual prognosis of these patients. Further prospective controlled studies are necessary to confirm these data.