DNA指纹技术应用于培养表皮细胞异体移植后的判定

我们采用ＤＮＡ指标技术，成功地对２例烧伤病人供皮创面移植异体培养表皮后的４个活检标本进行了检测，结果证实培养表皮异体移植后至少可存活２８天，提示培养表皮异体移植后存活时间延长。     

聚合酶链反应与培养表皮细胞异体移植结果的判定

培养的人表皮细胞异体移植后究竟能否长期存活，因无客观的方法证实，目前争议较大。作者采用三种聚合酶链反应体外ＤＮＡ扩增技术（ＰＣＲ），对培养表皮细胞异体移植于不同创面的存活情况进行了判定。结果在１６例、２７个活检标本中均查到了供者细胞的遗传标记，其中最长一例为移植后９２天。这表明人培养表皮异体移植后可长期存活。作者认为三种ＰＣＲ扩增技术各有其优点及不足，根据具有情况配合作用，可对人培养表皮细胞异体移     

DNA指纹技术应用于培养表皮细胞异体移植后的判定

我们采用ＤＮＡ指纹技术，成功地对２例烧伤病人供皮创面移植异体培养表皮后的４个活检标本进行了检测，结果证实培养表皮异体移植后至少可存活２８天，提示培养表皮异体移植后存活时间延长。     

DNA指纹技术应用于培养表皮细胞异体移植后的判定

我们采用DNA指纹技术,成功地对2例烧伤病人供皮创面移植异体培养表皮后的4个活检标本进行了检测,结果证实培养表皮异体移植后至少可存活28天,提示培养表皮异体移植后存活时间延长。     

复合皮移植后皮片生长和收缩情况的监测

目的观察不同复合皮移植后皮片的生长和收缩情况。方法对异体乳猪真皮＋自体表皮（组１），异体小猪真皮＋自体表皮（组２）和自体表皮（组３）共３个组移植的皮片，结合皮片完整性和猪妊娠过程等因素，经２年多动态监测移植皮片面积。结果移植后２～１２４周复合皮表面光滑无瘢痕；３６周后３组皮片面积均有不同程度生长扩大，依次为组１＞组２＞组３（Ｐ＜０．０１）；在复合皮组，２４周后整张皮片明显优于拼接皮（Ｐ＜０．００１）；猪妊娠过程显示，复合皮组有较好的组织增生能力和弹性回缩力（Ｐ＜０．０１）；组织学观察组１乳头和皮钉等组织结构成熟早，２４周时已接近正常皮肤。结论经酶处理的冷冻异体乳猪真皮复合移植后具有良好的组织弹性、耐磨性和细胞生长能力，是理想的真皮替代物；实验动物妊娠可客观地用于复合皮质量的检测。     

复合皮移植后皮片生长和收缩情况的监测

目的观察不同复合皮移植后皮片的生长和收缩情况。方法对异体乳猪真皮 自体表皮(组1),异体小猪其皮 自体表皮(组2)和自体表皮(组3)共3个组移植的皮片,结合皮片完整性和猪妊娠过程等因素,经2年多动态监测移随皮片面积。结果移植后2～124周复合皮表面光滑无瘢痕;36周后3组皮片面积均有不同程度生长扩大,依次为组1>组2>组3(P<0.01);在复合皮组,24周后整张皮片明显优于拼接皮(P<0.001);猪妊娠过程显示,复合皮组有较好的组织增生能力和弹性回缩力(P<0.01);组织学观察组1乳头和皮钉等组织结构成熟早,24周时已接近正常皮肤。结论经酶处理的冷冻异体乳猪真皮复合移植后具有良好的组织弹性、耐磨性和细胞生长能力,是理想的真皮替代物;实验动物妊娠可客观地用于复合皮质量的检测。     

客观判定人培养表皮细胞同种移植后存活情况的方法

培养表皮细胞同种移植后是否能长期存活,目前国内外均有争议,为明确培养表皮同种移植后的转归,国内外已有人采用免疫细胞化学技术及分子生物学技术对这一问题进行了研究。本文综述了到目前为止已采用的方法及结果。     

复合皮移植后皮片生长和收缩情况的监测

OBJECTIVE: To evaluate the growth and contraction of different composite skin grafts. METHODS: The status of the following composite skin grafts was evaluated over 2 years: allogeneic sucking pig dermis and autologous epidermis (group 1, n = 19), allogeneic young pig dermis and autologous epidermis (group 2, n = 7), autologous epidermis (group 3, n = 8). The effects of gestation on the composite skin grafts were also assessed. RESULTS: It showed that composite skin grafts had smooth surface and no scar formation during 2 to 124 weeks after transplantation. After week 36, the surface area of each group enlarged with different growth rates: group 1 > group 2 > group 3 (P < 0.01). Whole piece grafting was significantly better than joining together of smaller pieces after week 24 (P < 0.001). During pregnancy, CS had good growth ability and elasticity (P < 0.01). Histological study showed that group 1 had rapid growth of skin papillae and epidermal ridges similar to normal skin at week 24. CONCLUSION: As CS graft with trypsin treated allogeneic sucking pig dermis as a substitute for dermis showed the advantages of rapid growth, good elasticity and stability, it is suggested that it could be used in clinical practice.     

自异体皮混合移植成活机制中角质形成细胞及细胞因子作用的实验研究

目的　观察细胞因子及角质形成细胞在自异体皮混合移植成活机制中的作用。　方法建立自异体皮混合移植大鼠模型和人体表皮细胞 淋巴细胞混合培养 (MELC)模型 ,检测大鼠移植术后血清及创面局部组织标本培养上清液中白细胞介素 (IL)1 0的水平 ,观察体外表皮细胞在诱导免疫抑制中的作用。　结果　术后 7d,移植大鼠血清IL 1 0含量为 (2 5 .89± 2 .82 )ng/L,明显高于正常大鼠的 (1 4 .2 0± 2 .4 3)ng/L(P <0.0 5 )。术后 4～ 1 4d,大鼠组织标本培养上清液中IL 1 0含量显著不同(P <0.0 5或 0 .0 1 ) ;术后 2 1、2 8d与正常大鼠比较 ,差异无显著性意义 (P >0.0 5 )。自体表皮细胞、角质形成细胞在MELC体系中的抑制作用与其剂量相关。自体角质形成细胞加入到MELC中 ,辅助性T淋巴细胞 (Th)1分泌的细胞因子谱主要通过IL 1 0介导而抑制Th2细胞因子谱的分泌 ;当加入抗IL 1 0单克隆抗体后该抑制作用消失。　结论　自异体皮混合移植后 ,嵌入自体皮片中的角质形成细胞通过激活Th2使局部IL 1 0水平升高 ,这可能是移植成活的重要原因之一。     

异体腱细胞的分离培养与鉴定

异体腱细胞的分离培养与鉴定康庆林，田万成，王爱民如何最大限度激发腱细胞参与肌腱愈合是解决肌腱粘连的关键。体外培养是研究腱细胞生物学特征的重要手段，国外早在７０年代就进行了动物腱细胞培养，但有关异体腱细胞培养报道较少，我们成功地用贴块法培养了异体腱细胞...     

Activation of Tim-3-Galectin-9 pathway improves survival of fully allogeneic skin grafts

T cell immunoglobulin and mucin domain (Tim)-3 is a molecule expressed on terminally differentiated murine Th1 cells but not on Th2 cells. Identification of Galectin-9 as a ligand for Tim-3 has now firmly established the Tim-3-Galectin-9 pathway as an important regulator of Th1 immunity, which results in apoptosis of Th1 cells. Here, we demonstrate that engagement of Tim-3 by mouse recombinant Galectin-9 remarkably suppresses allograft rejection and improves survival of allogeneic skin grafts. Furthermore, administration of recombinant Galectin-9 decreases Tim-3 positive cells in draining lymph node and selectively inhibits production of IFN-gamma after skin transplantation. At last, even low dose of Galectin-9 (1 microg/ml) can obviously inhibit TCR crosslinking-induced primary T cell proliferation in vitro. These observations suggest that Tim-3-Galectin-9 pathway plays an important role in the termination of productive Th1-immune response and could lead to developing novel therapies in transplant medicine.     

The synergistic effect of low-dose cyclosporine and fluocinolone acetonide on the survival of rat allogeneic skin grafts

Both CsA and topical FA can prolong the survival of skin allografts under the proper conditions. This study was performed to determine if there is a synergistic effect between these two compounds. Buf (RT1b) rat split-thickness skin grafts were transplanted onto the backs of Lew (RT1l) rats. The MST for the control group was 9.89 +/- 0.35 days. In rats given oral CsA, 2.5 or 5 mg/kg, daily from the second day of grafting, the MST was 16.0 +/- 1.9 and 13.6 +/- 0.4 days (blood CsA levels were 166 +/- 20 and 640 +/- 32 ng/ml at the time of rejection, respectively. Topical FA applied daily beginning 72 hr after grafting resulted in a MST of 24.1 +/- 3.6 days. When both topical FA and 5 mg/kg oral CsA were used, the allograft survival time was more than 100 days in 4 of 7 animals. When oral CsA 2.5 mg/kg was combined with topical CsA and FA, the allograft rejection was delayed until 50 days postgrafting in four of six animals. The synergistic effects of oral CsA and topical FA is significant, and thus allows for the use of a subtherapeutic dosage of each compound and provides a potentially safe means for prolonging skin allograft survival.     

Early vascularization of syngeneic, allogeneic and xenogeneic skin grafts

The vascularization of syngeneic, allogeneic and xenogeneic skin grafts 3-4 days after grafting was studied in inbred rats by injecting hosts with 51Cr labelled red cells on the third day and counting graft radioactivity on the fourth day. Vascularization as determined by this method was quantitatively similar in syngeneic, allogeneic and mammalian xenogeneic grafts, while the vascularization of non-mammalian xenografts was significantly inferior. For any one donor-host combination, full-thickness grafts contained relatively more blood than did split-thickness grafts, presumably because of the large vessels in the deep part of the dermis in full-thickness grafts. Full-thickness neonatal grafts were vascularized similarly to full-thickness adult skin grafts. Grafted skin contained relatively more blood than did the corresponding normal skin. Reconstituted freeze dried allogeneic skin grafts contained virtually no blood, a phenomenon possibly analogous to the 'no reflow' phenomenon of microsurgery. The inferior vascularization of non-mammalian xenografts is more plausibly explained on the basis of defective self recognition than as representing a reaction to foreign determinants. These results do not necessarily indicate that one type of graft is better than another in clinical practice. But if the beneficial effects of temporary skin grafts do indeed depend on their capacity to become vascularized, fresh skin appears preferable to reconstituted freeze dried skin.     

Surgical management of hands in children with recessive dystrophic epidermolysis bullosa: use of allogeneic composite cultured skin grafts.

Recessive dystrophic epidermolysis bullosa (RDEB) is characterised by progressive childhood hand syndactyly and flexion contractures, which can be managed surgically but require split thickness autografts to facilitate satisfactory postoperative healing. We report on the partial substitution, for autografts, of improved composite cultured skin (CCS) allografts. The structure and preparation of these CCSs is outlined and their application in the course of 16 operations performed on 7 RDEB children with syndactyly and flexor contractures of fingers is described. Hand contractures were released and web spaces were covered with local flaps and split thickness autografts, while adjacent sides of the digits and other areas, as well as donor sites were generally grafted with CCS. Morphologic and functional results with CCS were judged to be good to excellent, the average time to recurrence was increased approximately 2-fold and smaller autografts needed to be used. In addition, healed CCS-treated donor sites could provide superior donor sites for further surgery.