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.     

移植骨髓间充质干细胞对兔退变椎间盘髓核细胞凋亡的影响

目的 观察骨髓间充质干细胞(MSCs)移植对兔退变椎间盘髓核细胞凋亡的影响.方法 以各兔L2/3、L3/4、L4/5、L5/6节段分为正常组、退变组、成纤维细胞(SFs)移植对照组、MSCs移植治疗组.MSCs和SFs分别经绿色荧光蛋白(GFP)转染后,注射植入退变椎间盘的髓核.通过透射电镜观察退变椎间盘凋亡髓核细胞形态;用实时定量聚合酶链反应(PCR)检测退变组织中髓核细胞凋亡相关基因bcl-2和box mRNA的表达;免疫荧光法标记髓核细胞凋亡相关蛋白Caspase-3,并通过TUNEL法标记凋亡髓核细胞,激光共聚焦显微镜检测髓核细胞凋亡蛋白表达率和细胞凋亡比率.结果 透射电镜下,退变椎间盘中凋亡髓核细胞呈现出核染色质边集,空泡形成,核膜断裂,凋亡小体形成等变化.MSCs移植治疗组bcl-2 mRNA的表达量高于退变组和SFs移植对照组(P<0.05),bax mRNA的表达量与退变组差异无统计学意义(P>0.05).MSCs移植治疗组细胞凋亡率和Caspase-3表达率均高于正常组[细胞凋亡率分别为(16.75±2.14)%和(6.86±1.08)%;Caspase-3表达率分别为[(20.34±1.03)%和(6.09±0.77)%](P<0.05),低于退变组和SFs移植对照组[细胞凋亡率分别为(31.87±4.16)%和(29.02±2.16)%;Caspase-3表达率分别为(31.50±3.78)%和(30.20±4.93)%](P<0.05).结论 髓核细胞凋亡在椎间盘退变过程中起重要作用.MSCs移植能有效抑制椎间盘髓核细胞凋亡,延缓椎间盘退变过程.     

绿茶多酚改善环孢素A抑制血管舒张的作用

目的 观察绿茶多酚(GTP)对环孢素A(CsA)抑制血管舒张作用的改善,并探讨其机制.方法 将30只SD大鼠随机平均分为3组:CsA组、对照组和CsA+GTP组.建模5周后,检测体质量、肾功能:血尿素氮(BUN)、肌酐(Cre);并取胸主动脉环,观察乙酰胆碱(Ach)诱发的血管舒张反应、左旋硝基精氨酸甲酯(L-NAME)和吲哚美辛预处理对舒张反应的影响、去内皮细胞的血管舒张反应.并检测血管组织一氧化氮(NO)水平.结果 实验5周后,CsA组大鼠体质量(253.2±8.1)g低于对照组(292.1±9.5)g;CsA+GTP组体质量(287.9±9.7)g高于CsA组;CsA组BUN、Cre含量高于对照组;CsA+GTP组BUN、Cre低于CsA组;差异均有统计学意义(P<0.05).CsA组大鼠Ach引起的动脉环的最大舒张度为(42.5±4.3)%,低于对照组的(81.2±7.6)%和CsA+GTP组的(70.1±6.5)%,差异有统计学意义(P<0.05).用L-NAME预处理后,CsA组和CsA+GTP组动脉环的舒张幅度分别为(40.3±3.7)%和(45.8±4.2)%,均低于对照组的(79.4±6.8)%;用吲哚美辛预处理后对照组和CsA+GTP组的舒张反应高于CsA组;差异均有统计学意义(P<0.05).去内皮细胞的各组血管中,血管舒张反应被明显抑制,各组血管的舒张百分比差异无统计学意义(P>0.05).CsA组大鼠血管组织中NO含量显著低于对照组和CsA+GTP组,差异有统计学意义(P<0.05).结论 CsA可导致血管组织NO水平下降,引起NO介导的内皮依赖性血管舒张功能异常.应用绿茶多酚后,则升高其血管组织中NO水平,改善内皮依赖性的血管舒张功能.     

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.     

表达增强型绿色荧光蛋白的骨肉瘤细胞亚株建立及生物学特性

目的 建立经绿色荧光蛋白基因转染的骨肉瘤细胞亚株并研究其生物特性。方法 利用脂质体转染增强型绿色荧光蛋白真核表达质粒（pEGFP-N1）人人骨肉瘤MG63细胞系，通过有限稀释法和细胞电泳，获得两株细胞克隆M6和M8，经体外细胞增殖、软琼脂形成、生长曲线、裸鼠成瘤试验综合分析其生物学行为改变。应用组织形态学观察、染色体分析、软琼脂克隆形成法研究癌细胞的生物学特性。结果 M6和M8两株在细胞电泳率和侵袭性上差异有统计学意义（P〈0．05），其中M6的群体倍增时间为38．4h，软琼脂形成率为18．7％，M8群体倍增时间为23．0h，软琼脂形成率为29．3％。裸小鼠背部皮下接种M6和M8，发现M8成瘤时间短，细胞增殖快，但在4周内两者均不发生转移。结论 骨肉瘤细胞亚系有不同的转移特性，GFP的整合及表达未对MG63细胞的生长状态造成明显影响，可作为报告基因进一步了解骨肉瘤细胞转移的差异性分析。     

骨髓干细胞参与小鼠急性肾小管坏死修复的实验研究

【摘要】 目的 观察在生理和病理情况下,骨髓来源的干细胞(BMSC)能否分化成肾小管上皮细胞。 方法 绿色荧光蛋白(GFP)标记的C57BL/6转基因小鼠提供骨髓细胞。同种无荧光标记的C57BL/6小鼠100只分为正常对照组、全身照射组、缺血再灌注组、骨髓移植组、骨髓移植+缺血再灌注组。受体鼠的骨髓重建经血液常规检查和流式细胞仪检测确认,并采用荧光组织化学、免疫组织化学等方法观察绿色荧光标记的BMSC在受体鼠肾脏的分布及数量。 结果 全身致死剂量γ射线照射未造成小鼠肾脏组织结构和生理功能的明显改变。骨髓移植后第56、84天的受体鼠肾小管中有少量GFP阳性细胞的存在[(78.75±5.99)%、(79.58±4.60)%],激光共聚焦显微镜进一步证实这些细胞位于肾小管,并表达肾小管上皮细胞特异性的功能蛋白megalin。 结论 在生理和病理情况下,骨髓干细胞均可以向肾小管上皮细胞转分化,参与肾小管上皮细胞的更新,并且在急性肾小管坏死的病理条件下,骨髓干细胞的肾向转化率与肾脏受损程度有关。     

The in vitro effect of antimicrobial photodynamic therapy with indocyanine green on Enterococcus faecalis: Influence of a washing vs non-washing procedure

IntroductionThe purpose of this study was to evaluate the in vitro effect of washing and non-washing of indocyanine green (ICG) as photosensitizer (PS) on bacterial count, biofilm formation, development and degradation of Enterococcus faecalis.MethodsThe anti-bacterial, anti-biofilm formation, anti-biofilm development and biofilm degradation of anti-microbial photodynamic therapy (aPDT) against E. faecalis was determined at concentrations of 3 to 2000 μg/mL of ICG, subject to 18 J/cm² dose of diode laser (808 nm) in washing and non-washing producers. Bacterial viability measurements and biofilm assays were evaluated by broth microdilution method and crystal violet assays, respectively.ResultsICG-mediated aPDT, using 25 to 2000 μg/mL and 50 to 2000 μg/mL showed significant reduction in E. faecalis growth when compared to the control in non-washing and washing producers, respectively (P < 0.05). Also, ICG-mediated aPDT showed a significantly inhibitory effect on biofilm formation of E. faecalis in concentration of 6 to 2000 μg/mL and 100 to 2000 μg/mL in non-washing and washing groups (P < 0.05). The biofilm development was inhibited by concentrations of 12 to 2000 μg/mL and 100 to 2000 μg/mL in non-washing and washing groups. The biofilm degradation increased from concentrations of 12 to 2000 μg/mL and 250 to 2000 μg/mL in non-washing and washing groups, respectively.ConclusionThis study shows that the application of ICG should be accompanied by laser irradiation without being washed out to achieve better result for bacterial count reduction and anti-biofilm effects.     

脑内移植GFP基因修饰细胞的研究

目的为确定外源GFP基因能否作为标记基因来追踪细胞在植入脑内后的生长情况。方法构建GFP基因真核表达质粒,导入PA317细胞,筛选出GFP高表达的PA317细胞克隆,移植入SD大鼠纹状体,作相应部位的组织冰冻切片,用荧光显微镜观察GFP在脑内的表达。结果GFP高表达的PA317细胞克隆,发出强绿色荧光,被移植入SD大鼠纹状体后．仍发出绿色荧光。结论GFP基因可以作为标记基因追踪细胞在植入脑内后的生长情况。