地拉罗司对大鼠窄蒂缺氧皮瓣模型促进微血管生成的实验

目的 探索术前应用地拉罗司(deferasirox)对大鼠乒乓拍样狭长窄蒂缺氧皮瓣模型微血管生成的促进作用及与用药时间的相关性.方法 成年雄性SD大鼠40只,随机分为对照组(8只),实验组(32只,分为1、3、5、7d组,每组8只).实验组:地拉罗司100 mg/kg分别连续灌胃1、3、5、7d;对照组:生理盐水100 mg/kg连续灌胃7d,灌胃结束后,次日行乒乓拍样窄蒂任意型皮瓣成形术,均于术后第7天取材,应用免疫组织化学、Western印迹及实时荧光定量聚合酶链式反应(PCR)法检测CD34表达,应用实时荧光定量PCR检测低氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的表达.结果 实验组较对照组皮瓣成活率明显提高,随着用药时间的增加,皮瓣微血管密度(MVD)、HIF-1α和VEGF均逐渐增高,7d达最高.结论 地拉罗司能诱导HIF-1α的分泌,促进VEGF产生,增加MVD表达,明显具有提高皮瓣对低氧的耐受,减少内皮细胞低氧后的坏死率,对缺血缺氧的窄蒂皮瓣具有较好的保护作用.     

地拉罗司通过诱导上皮细胞间质化改变促进窄蒂皮瓣微血管生成的实验研究

目的 探讨地拉罗司能否通过诱导上皮细胞发生间质化改变,从而促进皮瓣微血管生成.方法 雄性成年SD大鼠32只随机均分为Ⅰ组和Ⅱ组,2组又分别分为a、b组,每组各8只.Ⅰa组连续7d灌胃地拉罗司100 mg/kg,Ⅰb组连续7d灌胃等量生理盐水,灌胃结束后,次日行大鼠背部窄蒂皮瓣成形术,术中观察皮下微血管情况,术后第7天观察皮瓣愈合情况,并计算皮瓣成活面积百分比.Ⅱa组连续7d灌胃地拉罗司100 mg/kg,Ⅱb组连续7d灌胃等量生理盐水,灌胃结束后,次日行大鼠背部窄蒂皮瓣成形术,术后第3天皮瓣远端采集标本,免疫组化和Western blot法检测分化簇因子34(C D34)、E钙黏素、波形蛋白的含量,并计算皮瓣微血管密度.结果 Ⅰa组皮瓣下微血管生长较Ⅰb组旺盛.Ⅰa组皮瓣存活率100％,Ⅰb组存活率62.5％ (P ＜0.05);Ⅰa和Ⅰb皮瓣成活面积百分比分别为(100±0.OO)％和(84.06±4.42)％(P＜0.05).免疫组化及Western blot法检测示lⅡa组E钙黏素低于Ⅱb,而波形蛋白、CD34的表达量高于Ⅱb(P＜0.05).结论 地拉罗司能够通过诱导上皮细胞发生间质化改变促进皮瓣组织微血管生成,具有较强的促进窄蒂皮瓣愈合的作用.     

缺氧缺血型狭长窄蒂皮瓣成活过程中SDF-1表达的实验研究

目的 检测基质细胞衍生因子-1( stromal cell derived factor1,SDF-1)在缺氧缺血型狭长窄蒂皮瓣组织不同时间、不同面积的表达,研究SDF-1/CXCR4轴在狭长窄蒂皮瓣组织血管新生中的作用.方法 在5头家猪双侧背部均制作蒂部长宽比例为4:2,携带面积分别为2 cm ×2 cm(A组皮瓣)、3 cm × 3 cm(B组皮瓣)、4 cm×4 cm(C组皮瓣)、5 cm × 5 cm(D组皮瓣)和6 cm ×6 cm(E组皮瓣)的狭长窄蒂皮瓣模型.对5组皮瓣进行大体观察、成活面积计算、皮瓣组织HE染色,用ELISA法检测皮瓣组织远端在术中及术后3、5、7、14 d时SDF-1表达变化情况.结果 ①同一组狭长窄蒂皮瓣,随着时间推移,SDF-1的表达也随之增加,在第5天达到最高值,随后逐渐下降,趋于基础水平.②不同组狭长窄蒂皮瓣,随着皮瓣面积的增大,SDF-1的表达也随之增加,当皮瓣面积达5cm×5 cm时,SDF-1的表达量不再增加,皮瓣远端部分坏死.结论 SDF-1的表达量可能与狭长窄蒂皮瓣成活过程中的供血量相关.     

缺氧缺血型狭长窄蒂皮瓣成活过程中基质细胞衍生因子-1、CD34及微血管密度的表达

目的检测基质细胞衍生因子-1（stromal cell—derived factor-1,SDF-1）、CD34、微血管密度（MVD）在缺氧缺血型狭长窄蒂皮瓣组织的不同时间、不同面积的表达,以及SDF-1在狭长窄蒂皮瓣组织血管新生中的作用。方法在5头家猪两侧背部各制作蒂部长宽比为4：2,携带皮瓣面积分别为2cm×2cm（A组）、3cm×3cm（B组）、4cm×4cm（C组）、5cm×5cm（D组）和6cm×6cm（E组）共5组的窄长蒂皮瓣模型。对5组皮瓣进行大体观察、皮瓣成活情况、光镜下检测MVD、免疫组织化学及酶联免疫技术（ELISA）,检测皮瓣远端组织在术后0、3、5、7、14d时,SDF-1和CD34表达。结果（1）同一组狭长窄蒂皮瓣,随着时间推移,SDF-1、CD34及MVD表达也随之增加,SDF-1在第5天达最高值（A组124．80±4．05,B组137．85±3．03,C组166．53±2．98,D组72．80±2．63,E组62．79±2．20）;CD34在第7天达最高值（A组16．76±0．62,B组17．60±0．72,C组18．48±0．55,D组12．70±0．60,E组11．51±0．70）;MVD在第7天达最高值（A组52．45±2．78,B组59．34±3．12,c组61．14±3．35,D组25．25±3．78,E组24．46±7．46）,随后逐渐下降。（2）不同组狭长窄蒂皮瓣,随着皮瓣面积的增大,SDF-1、CD34及MVD的表达也随之增加,当面积达5cm×5cm时,各项指标不再增加,皮瓣远端出现部分坏死。结论SDF-1对狭长窄蒂皮瓣成活中的新生血管起一定的调节作用。     

一氧化氮合酶和CD34在狭长窄蒂随意型皮瓣中表达与皮瓣成活的关系

目的通过在猪背部设计一定宽长比例的狭长窄蒂所携带不同大小的几组随意型皮瓣,在术后不同时间测定各组皮瓣内一氧化氮合酶（nitri coxide synthase,NOS）和CD34的表达量,并观察皮瓣的成活面积,揭示NOS、CD34表达量与皮瓣成活的关系,以探讨狭长窄蒂皮瓣的成活机制。方法在5只白色实验猪的两侧背部均形成蒂部宽长比皆为2：4的狭长窄蒂皮瓣各5个,其面积分别为2cm×2cm、3cm×3cm、4cm×4cm、5cm×5cm、6cm×6cm,并依次命名为A、B、C、D、E皮瓣。其中A瓣为对照瓣。术后对皮瓣进行大体观察,并分别于术中即刻、术后3、5、7、14d切取皮瓣远端外侧缘组织块,用于测定皮瓣内NOS、CD34表达量,术后14d计算皮瓣存活面积。结果A、B、C组皮瓣NOS、CD34的表达量随皮瓣面积的增大逐步升高,皮瓣完全存活（P〈0．05）;D、E组皮瓣当皮瓣面积达到一定限度时,NOS、CD34表达餐不再随皮瓣面积的增大而升高,表达相对恒定,皮瓣远端部分坏死（P〉0．05）。结论任意型皮瓣的成活与蒂的宽长比没有直接关系。一定宽长比例的狭长窄蒂所携带的随意皮瓣成活面积有其一定限度。     

一定长宽比狭长窄蒂与任意皮瓣成活面积关系的实验研究

目的 揭示具有一定长宽比例的狭长窄蒂与任意皮瓣成活面积关系. 方法 25头猪被随机分成5组,每组5头,5组皮瓣蒂长宽比分别为0 cm:2 cm、1 cm:2 cm、2 cm:2 cm、3 cm:2 cm、4 cm:2 cm,每个长宽比例的狭长窄蒂均携带5个不同面积的任意皮瓣,分别为2 cm×2 cm、3 cm×3 cm、4 cm×4 cm、5 cm×5 cm和6 cm×6cm,并依次命名为A、B、C、D、E,其中A瓣为B、C、D、E瓣的对照瓣,在每组每只猪的双侧背部均形成A、B、C、D、E皮瓣,顺序随机排列.对每组皮瓣进行大体观察、荧光色素钠染色、ECT血流测定、成活面积分析等. 结果 ①当狭长窄蒂的长宽比例一定时,随着皮瓣面积的增加,皮瓣成活面积也随之增大,但达一定界限时皮瓣远端即发生坏死,而成活面积并未缩小;②当皮瓣大小一定,随着狭长窄蒂的长宽比例增加,皮瓣成活面积不受影响,但达一定界限时皮瓣远端即发生坏死,皮瓣成活面积缩小. 结论 任意皮瓣的蒂部可以设计成狭长状,蒂宽可远远小于瓣宽;一定长宽比的狭长窄蒂所能携带的任意皮瓣成活面积有其最大值,一定范围内增大皮瓣面积或蒂部的长宽比例不会导致皮瓣坏死.     

狭长窄蒂皮瓣在面额部皮肤癌治疗中的应用

目的 介绍一种打破常规任意皮瓣长宽比例限制的狭长窄蒂皮瓣在面额部皮肤癌切除后组织缺损修复中的应用,并探讨其成活机制.方法 设计不带知名血管,以耳前或耳后狭长皮肤筋膜或单纯筋膜为蒂的侧颌颈部皮瓣,修复面额部皮肤癌切除后组织缺损26例,其中颞额区皮肤基底细胞癌5例,皮肤鳞状细胞癌6例,面部恶性黑色素瘤1例,基底细胞癌8例,鳞状细胞癌5例,皮肤黏液癌1例.26例皮瓣的蒂部有24例在耳前,2例在耳后;4例仅为筋膜蒂.皮瓣最大10.0 cm×8.0 cm,最小3.0 cm×2.5cm,蒂宽1.0～1.5 cm,蒂长2～6 cm.结果 26例狭长窄蒂皮瓣除5例皮瓣远端淤血,其后渐恢复,其余全部成活,修复后局部外形理想.结论 该狭长窄蒂皮瓣不带知名血管,其长宽比例超过常规任意皮瓣,无需解剖血管,切取容易,蒂部狭长使其旋转和覆盖范围大,供、受区邻近,是修复面额部皮肤癌扩大切除后组织缺损的理想皮瓣.     

狭长窄蒂皮瓣在老年人皮肤癌治疗中的应用

目的 探讨狭长窄蒂皮瓣在老年肿瘤切除后软组织缺损修复中的应用及成活机制.方法 设计蒂部位于肿瘤切除后创面旁沿知名或穿支血管走向的狭长窄蒂皮瓣,用于老年人肿瘤切除后软组织缺损的修复.皮瓣切取最大面积12 cm×10 cm,最小3 cm×4 cm,蒂宽1.0～1.5 cm,蒂长2～8 cm.结果 共17例,14例Ⅰ期成活,外形良好,3例因早期包扎不当,延迟愈合.结论 老年人皮肤组织细胞代谢率低,移植后皮瓣组织本身的耐缺血缺氧能力强,设计狭长窄蒂皮瓣修复老年肿瘤切除后软组织缺损,较易成活,且窄蒂狭长,转移容易,皮瓣不臃肿,同时利于肿瘤的早期综合治疗,是老年肿瘤术后修复较理想方法.     

猪狭长窄蒂随意型皮瓣蒂部不同长宽比例与成活面积关系的研究

目的 探讨狭长窄蒂随意型皮瓣蒂的长宽比例与皮瓣成活面积的关系,为临床应用狭长窄蒂皮瓣提供理论依据.方法 将25只猪随机分成5组,每组5只,5组皮瓣蒂的长、宽比分别为0∶2,1∶2,2∶2,3∶2,4∶2,每个长宽比例的狭长窄蒂均携带5个不同面积的随意型皮瓣,分别为2 cm×2 cm、3 cm×3 cm、4 cm×4 cm、5 cm×5 cm、6 cm×6 cm,并依次命名为A、B、C、D、E,其中A瓣为B、C、D、E瓣的对照瓣,在每组每只猪的双侧背部均形成A、B、C、D、E皮瓣,顺序排列但两侧背部相反.对每组皮瓣进行大体观察、荧光色素钠染色、ECT血流测定、病理学表现和成活面积分析等.结果 ①狭长窄蒂随意型皮瓣和常规随意型皮瓣的成活过程和病理过程是一致的,皮瓣的成活过程并未因蒂部变窄变长而延缓.②当狭长窄蒂的长宽比例不变时,随着皮瓣面积的增加,皮瓣成活面积也随之增大,但达一定界限时皮瓣远端即发生坏死,而成活面积并未缩小.③当皮瓣大小不变,随着狭长窄蒂的长宽比例增加,皮瓣成活面积不受影响,但达一定界限时皮瓣远端即发生坏死,皮瓣成活面积缩小.结论 ①随意型皮瓣的蒂宽可远远小于瓣宽,蒂宽和瓣长比可远小于传统的比例.②随意型皮瓣的蒂部可以设计成狭长状,使整个皮瓣形似"乒乓球拍",更利于皮瓣的旋转.③不同长宽比的狭长窄蒂所能携带的随意型皮瓣成活面积有其最大值,一定范围内增大皮瓣面积或蒂部的长宽比例不会导致皮瓣坏死.

Abstract：

Objective To explore the relationship between the ratio of length to width of slender narrow pedicle and random flap survival area, and to provide a theoretical basis for the clinical application of slender narrow pedicle flaps. Methods 25 pigs were randomly divided into 5 groups, 5 pigs in each group. The ratio of length to width of slender narrow pedicle in 5 groups respectively was: 0∶2 , 1∶2 ,2∶2, 3∶2, 4∶2 . Every ratio's slender narrow pedicle was carrying five different size of random flaps,which were2cm×2cm(A),3 cm×3 cm(B),4 cm×4 cm(C), 5 cm×5 cm(D),6 cm×6 cm(E),respectively. Flap A was control flap. In each group, flap A, B, C, D and E were created in each pigs'bilateral back. The order in both sides back is contrary. The flaps were evaluated with the general observation, fluorescence examination, blood flow ECT test, pathological expression and computerized analysis of survival area. Results ① The living process and pathologic process of traditional flap and slender narrow pedicle flap were consistent. It could not postpone the flap living process when the flap pedicle became long and narrow. ② When the ratio of the length to width of the slender narrow pedicle was constant, along with the flap area increased, the flap survival area also increased, but when the flap reached a certain area , the distal flap would necrosis, the flap survival area would not reduce. ③ When the flap size remained unchanged, along with the ratio of the length to width of the slender narrow pedicle increased, the flap survival area was not affected, but when the ratio of the length to width of the slender narrow pedicle increased to a certain limit, distal flap would necrosis, the flap survival area would reduce.Conclusions ① Pedicle width of random flap can be much smaller than flap width. The ratio of pedicle width to flap length is far less than traditional ratio. ② The pedicle of random flap can be designed as slender shape, so that the whole flap looks like "pingpang bat ", which makes the narrow pedicle flap rotate easily. ③ A certain ratio of the length to width of a slender narrow pedicle has a maximum flap survival area, and increasing the flapsize or ratio of the length to width of a slender narrow pedicle in a certain extent will not lead to flap necrosis.     

猪背部乒乓球拍样任意型皮瓣成活的动物实验研究

目的 探讨乒乓球拍样任意型皮瓣成活面积与窄蒂长宽比例关系.方法 25只猪随机分成5组,在5组猪的背部分别形成不同长宽比例的狭长窄蒂和5个不同面积的任意型皮瓣.对每组皮瓣进行大体观察、荧光色素钠染色、单光子发射计算机断层仪（ECT）血流量测定、成活面积分析等.结果 当狭长窄蒂的长宽比例不变时,随着皮瓣面积的增加,皮瓣成活面积也随之增大,但达一定限度时皮瓣远端即发生坏死,而成活面积并未缩小;当皮瓣面积不变,随着狭长窄蒂的长宽比例增加,皮瓣成活面积不受影响,但达一定限度时皮瓣远端即发生坏死,皮瓣成活面积缩小.结论 狭长窄蒂任意型皮瓣是一种简便实用的任意型皮瓣,蒂部可以设计成狭长状,蒂瓣的长宽比远小于传统的比例,在一定范围内增大皮瓣面积或蒂部的长宽比例不会导致皮瓣坏死.     

Soluble Fas and Fas-ligand in bladder cancer in vitro and in vivo

Circulating soluble Fas (sFas) and expression of Fas-ligand on cancer cells are mechanisms of immune escape. The aim of the present study was to investigate expression and production of Fas and Fas-ligand on bladder cancer cell lines of different grade as a basic mechanism of their secretion in vivo. sFas and sFas-ligand serum levels of patients with different stage of bladder cancer were examined to determine the possible clinical use of these molecules as tumor markers. Bladder cancer cell lines RT4 (G1), RT112 (G1), T24 (G3) and SUP (G4) were analyzed by flowcytometry for Fas and Fas-ligand expression. To determine if the Fas-ligand gene is transcribed in these bladder cancer cell lines, RT-PCR was performed on mRNA extracted from these cell lines. Production of sFas and sFas-ligand was examined in cell culture supernatants of the cancer cells as well as in the serum of 62 patients with bladder cancer by a specific ELISA test. We demonstrate that Fas is expressed in similar levels on all human bladder carcinoma cell lines. In T24 (G3) and SUP (G4) cell lines we were able to detect the Fas-ligand protein, whereas no Fas-ligand protein could be found in RT4 and RT112 (G1) cells. Fas-ligand mRNA was expressed in all bladder cancer cell lines. Furthermore, all bladder cancer cell lines produce sFas but no sFas-ligand in spite of mRNA expression. The range of sFas levels in the serum of all patients with bladder cancer was large and did not show a correlation to the histopathological stage of bladder cancer. Although there is in vitro evidence that sFas and Fas-ligand play a role in bladder cancer, no correlation between the sFas and s Fas-ligand serum levels and the histopathological stage of bladder cancer could be found. Therefore, serum sFas and sFas-ligand have to date limited clinical relevance.     

Predominantly posterior instrumentation and fusion in neuromuscular and neurogenic scoliosis in children and adolescents

We studied the results in 46 patients with neuromuscular and neurogenic scoliosis (average age 13.5 years, range 6–19 years) who had had posterior fusion with a modified Luque technique between May 1985 and June 1992. The main criteria to recommend surgery were curve progression, loss of balance when sitting, control of the head and difficulties in wearing an external orthotic support. The mean preoperative curve was 63°, the postoperative value was 24°, representing a correction of about 62%. The average number of stabilized segments was 13. In 39 out of 46 patients, lumbosacral fixation was included in the construct. Failure of implants, pseudarthroses and major losses of correction in purely neuromuscular scolioses could be avoided by using rigid segmental fixation and a dorsolateral fusion with a mixture of autologous and allogenous bone. The scoliosis most difficult to influence was found to be Friedreich's ataxia. In Duchenne muscular dystrophy the best method of treatment was surgery performed as early as possible, i.e. at the time of loss of walking capacity in the case of a scoliosis exceeding 20° and with two consecutive X-rays proving curve progression. Analysis of our series does not confirm the morbidity and complication rates of previous studies.     

Culture and characterisation of human urothelium in vivo and in vitro

The aim of this study was to culture human urothelium and generate enough cells for subsequent reconstructive surgery. Using a modification of the Rheinwald-Green method for the routine culture of keratinocytes from patients with burns, we successfully cultured 91% of 57 biopsies from the renal pelvis, ureter, bladder and urethra of paediatric patients. The cells could be split one to three up to 9 times at 7–10 day intervals, giving a surface area of 1000 cm² after a 2 month culture period. Primary cultures could not be initiated in defined medium MCDB153, although cells initiated using the Rheinwald-Green method could subsequently be propagated in this medium. Cytokeratin patterns in vitro were similar to those in vivo in the expression of keratins 7, 18 and 19 (characteristic of simple epithelia) and keratin 13 (characteristic of non-cornified stratified epithelia). Cultured urothelium also expressed keratin 14 (characteristic of cornified stratified epithelium) in about 25% of cells and keratin 16 (characteristic of fast-growing cells). These findings indicate that urothelial cells can be propagated in vitro for autologous grafting, and the next step is to identify substrates suitable for urothelial cell growth and differentiation and surgical manipulation.     

Overweight in childhood and bone density and size in adulthood

Summary  

We evaluated the adult bone structural traits in relation to childhood overweight in 832 men and women. Childhood overweight was associated with larger cross-sections at long bones in both sexes. Excess weight in childhood may also lead to higher trabecular density in females and somewhat lower cortical density in men.