VEGF gene therapy for the survival of transplanted fat tissue in nude mice.

The objective of this study was to determine the effects of adenovirus-mediated vascular endothelial growth factor (Ad-VEGF) on the angiogenesis and survival of free-fat tissue transplantation in nude mice. Thirty 6-week-old CD-1 nude male mice were injected with 1ml fat tissue (harvested by suction-assisted lipectomy from the breast of humans) in the subcutaneous of scalp and were randomised into three groups of 10 animals each. Group 1 was the study group, in which Ad-VEGF was mixed with transplanted fat tissue and injected into mice. In group 2, adenovirus-mediated green fluorescent protein (Ad-GFP) gene was mixed with transplanted fat tissue and injected into the mice. In group 3, normal saline alone was used. Both group 2 and group 3 are control groups. The animals were euthanised 15 weeks after the procedure. The fat survival weight and volume of the study group were significantly greater than those of two control groups (p<0.05). Light microscopical examination of haematoxylin and eosin-stained slides of the dissected fat 15 weeks after injection was performed in group 1 and group 2. Less cyst formation and fibrosis, indicating improved quality of the injected fat, can be obtained by the addition of Ad-VEGF. Vascular density was evaluated at the microvascular level through the use of light microscopic sections of the central part of the fat tissue at 15 weeks after injection by von Willebrand factor staining. Histological evaluation showed that capillary density increased markedly in the study group mice. Mice of the study group disclosed significantly higher VEGF protein levels detected by ELISA assay of plasma samples obtained from the mice after the fat injection (day 1, 4, 7 and 28; p<0.01) at each time point than the mice of the two control groups. The findings reported in this study indicate that the VEGF gene therapy can enhance the survival and the quality of grafted fat tissue, which may be due to induction of angiogenesis.     

血管内皮祖细胞移植提高裸鼠缺血皮瓣存活率

目的探讨血管内皮祖细胞(endothelialprogenitorcell,EPC)移植促进早期断蒂的缺血皮瓣的血管新生,从而提高皮瓣存活率的可能性。方法体外分离、培养人脐血中EPCs,免疫组织化学进行鉴定,然后移植于裸鼠随意皮瓣,皮瓣早期断蒂,裸鼠分两组:实验组(EPCs移植)、对照组(M199注射),术后皮瓣4d断蒂。观察皮瓣的存活率、激光多普勒血液监测仪监测血流灌注、CD34免疫组织化学检测皮瓣毛细血管密度、激光共聚焦检测EPCs在皮瓣内的密度。结果脐血中分离培养的EPCs表达CD34、KDR及CD133,实验组EPCs移植裸鼠皮瓣后,整合到缺血部位新生血管中,与对照组的皮瓣存活率分别为(60.3±2.1)%、(34.2±1.8)%(P<0.05);而且两组毛细血管密度、血流灌注差异均有统计学意义(P<0.05);实验组术后第7、11天时二组皮瓣中的EPCs密度分别为(75.2±6.5)个/mm2、(305.4±26.5)个/mm2,而对照组均为0个/mm2(P<0.05)。结论脐血中的EPCs体外培养后移植体内可促进缺血皮瓣的血管新生,提高存活率。     

The Role of Frozen Storage in Preserving Adipose Tissue Obtained by Suction-Assisted Lipectomy for Repeated Fat Injection Procedures

BACKGROUND: The injection of autologous free fat obtained by suction-assisted lipectomy for the correction of soft tissue defects is a common procedure in plastic surgery. However, unpredictable partial absorption of the injected fat often necessitates repeated procedures. OBJECTIVE: To examine the role of frozen storage as a means of preserving the fat obtained by suction-assisted lipectomy for repeated procedures. METHODS: Human adipose tissue obtained by suction-assisted lipectomy was stored in a domestic refrigerator at -18 degrees C for 2 weeks. After thawing, the fat was injected into nude mice. In the control group, the fat was injected immediately after the harvesting procedure. Grafts were dissected out and compared 15 weeks postinjection. RESULTS: Injected fat survived in both study and control groups. No significant differences were found between fat graft weight and volume, or in any of the histologic parameters examined. CONCLUSION: Fat obtained by suction-assisted lipectomy may be preserved for future use by freezing.     

MMP-2反义寡核苷酸抑制人膀胱癌裸鼠异体移植瘤生长的实验研究

目的：探讨基质金属蛋白酶2（MMP2）反义寡核苷酸对人膀胱癌裸鼠异体移植瘤生长的抑制作用。方法：制备膀胱癌裸鼠移植瘤模型18只,随机分为3组：MMP2反义寡核苷酸（ASODN）治疗组、MMP2正义寡核苷酸（SODN）治疗组及对照组,每组6只。待瘤结节直径≥5mm后,分别在肿瘤细胞接种部位周围及中心皮下注射反义寡核苷酸／阳离子脂质体复合物、正义寡核苷酸／阳离子脂质体复合物和生理盐水。每周2次,连续4周,断颈处死裸鼠,计算肿瘤体积,称量瘤重。常规切片,HE染色,观察组织学形态并进行病理学评估。应用免疫组织化学技术（SP法）检测各组移植瘤组织中PCNA蛋白表达。结果：与对照组瘤重（7．49±0．53）g比较,ASODN组瘤重（4．18±0．53）g明屁降低（P〈0．01）;ASODN组、正义寡核苷酸（SODN）组抑瘤率分别为44．19％、8．41％（P〈0．01）;ASODN组、对照组增殖指数（PI值）分别为27．63％、88．39％,抑制率为60．76％（P〈0．01）;ASODN组瘤体病理学特征改善。结论：MMP-2反义寡核苷酸能抑制裸鼠体内移植瘤生长,通过反义寡核苷酸下调MMP-2的表达,降低癌细胞的增殖活性,有效逆转肿瘤的恶性表型。为膀胱癌的基因治疗提供了实验依据。     

Effects of direct injection of dehydrated ethanol on PC3 human prostate cancer cells in nude mice: Preliminary study

OBJECTIVES: The effect of direct local injection of dehydrated ethanol on hormone-independent prostatic carcinoma cells (PC3 cells) implanted in nude mice was investigated. METHODS: PC3 tumors were implanted subcutaneously into 30 nude mice. Three weeks later, dehydrated ethanol was injected directly into the tumors. Twenty-three animals received an injection of ethanol at a volume of 40 microL, 80 microL, or 160 microL, and were divided into a low-dose group (n = 11) and a high-dose group (n = 12) on the basis of the ethanol/tumor volume ratio (<30% versus> or =30%). The control group (n = 7) was injected with 40 microL of physiological saline. The tumor volume before treatment was 324.9 +/- 110.9 mm(3) and was assessed as well as 1 day, 4 days, 1 week, 2 weeks, and 3 weeks after injection. Then the changes of tumor volume were compared between the two ethanol groups (low-dose group and high-dose group) and the control group. Histological examination was performed for up to 3 weeks after injection. RESULTS: Assessment of tumor volume showed that the ethanol/tumor volume ratio was 16.1 +/- 5.3% in the low-dose group (n = 5) and 51.8 +/- 20.3% in the high-dose group (n = 6). Tumor growth was significantly inhibited after 1 and 2 weeks in the ethanol groups compared with the control group (n = 3). After 3 weeks, there was a tendency for tumor regrowth in the low-dose group, but growth inhibition was maintained in the high-dose group. Histological examination showed tumor degeneration and necrosis with feeding vessel obstruction in the acute phase. CONCLUSIONS: Local injection of dehydrated ethanol regressed tumors of prostatic carcinoma cells in nude mice, with the degeneration of tumor cells and occlusion of feeding vessels.     

血管内皮生长因子和雌二醇促进血管内皮祖细胞分化生成血管的对比研究

目的：对比研究常规剂量下血管内皮生长因子（VEGF）和雌二醇对血管内皮祖细胞（EPCs）分化生成血管的促进作用。方法：分离培养人外周血EPCs,与基质胶混匀后注射到9只裸鼠双侧下腹部,另设2只注射等体积培养液与基质胶的混合液。将9只注射细胞的裸鼠随机分为3组,每组分别定期局部注射VEGF、雌二醇,生理盐水,定期观察记录血管组织块的生长状况。移植6周后取材,测量计算血管组织块的体积、HE染色观察血管组织块的血管增殖状况,各组之间进行对比。结果：给药组血管组织块体积与注射生理盐水组相比,具有显著性差异,体积明显偏大,而给药组之间体积未见显著性差异。HE染色观察可见各组的血管组织块内血管增殖明显,血管排列紊乱的,管腔大小不一。给药组血管密度明显大于注射生理盐水组,而给药组之间的血管密度差异较小。结论：VEGF和雌二醇组具有促进EPCs分裂增殖形成血管的能力,常规剂量下两者促进EPCs分裂增殖生成血管的能力无显著性差异。     

骨髓血管内皮祖细胞移植提高缺血皮瓣存活率的研究

目的：探讨大鼠骨髓来源的血管内皮祖细胞（endothelial progenitor cells,EPCs）移植促进缺血皮瓣的血管新生,从而提高皮瓣存活率的可能性。方法：在Wistar大鼠背部形成2cm×8cm蒂部位于双侧髂棘连线上的随意型超比例皮瓣,建立缺血皮瓣模型。采用密度梯度离心法从Wistar大鼠骨髓中分离出骨髓单个核细胞,并在专有的诱导培养基（EG-M2MV）中培养,贴壁筛选法分离,诱导分化为EPCs;对细胞的形态、表型、功能加以鉴定。将EPCs注射移植于iWstar大鼠背部的缺血皮瓣,大体观察皮瓣的存活率,vWF免疫酶染色法检测皮瓣毛细血管密度。以缺血皮瓣单纯注射磷酸盐缓冲液（PBS）的Wistar大鼠作为对照组。结果：骨髓中分离培养的EPCs呈现典型的＂铺路石＂样外观,表达CD34、Flk-1及CDl 33表型,能够摄取DIL-ac-LDL和特异性结合FITC-UEA-1。注射EPCs组的缺血皮瓣存活率以及毛细血管密度显著高于对照组（P〈0.05）。结论：骨髓中的EPCs在体外培养后注射移植于皮瓣内,可以促进缺血性皮瓣的血管新生,提高缺血性皮瓣的存活率。     

Transplantation of endothelial progenitor cells transferred by vascular endothelial growth factor gene for vascular regeneration of ischemic flaps

BACKGROUND: Neovascularization occurs through two mechanisms: angiogenesis and vasculogenesis. Therefore, there are two strategies to promote neovascularization: therapeutic angiogenesis and therapeutic vasculogenesis (endothelial progenitor cells therapy). MATERIALS AND METHODS: In this study, we examined whether or not endothelial progenitor cells combined with vascular endothelial growth factor (VEGF) gene therapy is useful for ischemia surgical flaps in vivo. At the same time, we quantitatively compared the neovascularization ability of transplanted endothelial progenitor cells (EPCs) transducted with VEGF165 gene and EPCs alone. EPCs were isolated from cord blood of healthy human volunteers, cultured in vitro for 7 days and identified by immunofluorescence. After transduced with VEGF165 gene in vitro, proliferative activity of EPCs was assessed using MTT assay. CM-DiI was used to trace EPCs in vivo 4 days after injection of 5 x 10(5) VEGF-transduced EPCs(VEGF-transduced EPCs group, n = 10), 5 x 10(5) EPCs (non-transduced EPCs group, n = 10) in 500 microL EBM-2 media, or 500 microL EBM-2 media (EBM-2 media group, n = 10) local, a cranially based flap was elevated on the back of nude mice. The percent flap survival, neovasculariztion and blood flow recovery of flaps was detected. RESULTS: EPCs expressed cell markers CD34, KDR, and CD133. A statistically significant increase in percent flap survival was observed in mice of VEGF-transduced EPCs group as compared with that of non-transduced EPCs group: 67.99 +/- 6.64% versus 59.43 +/- 4.69% (P < 0.01), and 41.24 +/- 2.44% in EBM-2 media group (P < 0.01). The capillary density and blood flow recovery of flaps in VEGF-transduced EPCs group were both improved. CM-DiI-labeled VEGF-transduced EPCs were observed in vivo and the numbers of cells increased. CONCLUSION: EPCs from human cord blood can increased neovascularization of ischemic flaps and augmented the survival areas, and VEGF-transduced EPCs have more powerful ability of promoting neovascularization in animal model of ischemic flaps.     

Immunoscintigraphy of xenotransplanted hepatoblastoma with iodine 131-labeled anti-alpha-fetoprotein monoclonal antibody.

BACKGROUND/PURPOSE: Hepatoblastoma (HB) is the most common primary malignant liver tumor affecting infants and young children. The alpha-fetoprotein level is elevated in 95% of all children with hepatoblastoma. Therefore, it is of interest to assess targeting of the HB marker alpha-fetoprotein by antibody imaging. In this pilot study, the authors investigated the radioimmunoscintigraphy of xenotransplanted HB in nude mice utilizing an anti-alpha-fetoprotein antibody. METHODS: HB cell suspensions from tumors of 3 children were transplanted subcutaneously into nude mice NMRI (nu/nu). A total of 200 microg of intact anti-alpha-fetoprotein antibody was injected intravenously into 8 animals from each HB. Before injection, the monoclonal antibody was labeled with iodine (I) 131 (specific activity of 75 MBq/mg, labeling yield of 95%) using the conventional iodogen method. Planar scintigraphic images of anesthetized mice in posterior views were acquired with a gamma camera immediately after injection, and after 1, 2, 3, 7, and 14 days. The biodistribution data were obtained by killing and dissecting animals, and the activity in the tissues was measured in a gamma counter. The alpha-fetoprotein levels in the animals' sera were recorded 15 days after imaging and were compared with the control group. RESULTS: A total of 66% of the hepatoblastomas could be detected by scintigraphy. Within 24 hours, the mean specific tumor uptake in nude mice hepatoblastomas with a volume of over 1,000 mm3, was 14% per injected dose (+/-3.9%). The biological half-life of the labeled antibody complex in the tumor was 3.86 (+/-0.84) days. Thyroid uptake of free I-131 was 2.85% per injected dose (+/-1.5%) reflecting the deiodination of the labeled antibody complex. CONCLUSIONS: The results show the possibility of imaging xenotransplanted hepatoblastoma with 131I-labeled anti-alpha-fetoprotein and may, in the future, determine tumor recurrence and extension, and thereby improve the prognosis of advanced HBs.     

人脐血内皮祖细胞治疗裸鼠心肌梗死

目的 探讨人脐血内皮祖细胞(EPCs)移植治疗裸鼠心肌梗死的可行性.方法 采用淋巴细胞分离液提取人脐血单个核细胞(MNCs),应用添加诱导因子的培养基于体外诱导分化并于培养7 d后进行鉴定.采用20只裸鼠建立心肌梗死模型后,将体外诱导分化7 d并摄取CM-Dil的内皮祖细胞通过尾静脉注射进行细胞移植到实验组,对照组注射培养基.2周后计数心梗区域新生毛细m管密度及心梗面积并于荧光显微镜下观察新生血管的荧光.结果 体外诱导7 d后贴壁细胞CD34阳性率达(50.48±5.17)%,CDl33阳性率达(19.12±4.37)%.实验组平均梗死面积为(8.27±1.64)%,对照组为(14.30±2.84)%(t=-4.78,P<0.05);实验组每高倍视野平均新生血管密度为14.29±1.38,对照组为10.17±1.72(t=4.71,P<0.01);行荧光显微镜下观察实验组新生血管有红色荧光.讨论人脐血单个核细胞在体外诱导分化为内皮祖细胞,进行细胞移植到建立心梗模型的裸鼠后可在心梗区域形成新生血管,从而并改善梗死部位心脏功能.     

人脂肪来源细胞体内构建组织工程化脂肪的实验研究

目的探讨以组织工程技术，应用脂肪来源细胞（Adipose—derived cells，ADCs）体内构建脂肪组织的可行性。方法吸脂术获得人脂肪组织，一部分直接植入裸鼠体内；另一部分用酶消化法分离、培养ADCs，将第三代细胞接种于纤维凝胶（Fibrin glue）支架中，经成脂肪培养液诱导1周后，植入裸鼠体内，4周后取材，用称重法及油红、HE染色检测结果。实验分为直接注射脂肪组、单纯支架组、细胞-支架复合体脂肪诱导组、非诱导组。结果直接注射组在裸鼠皮下形成脂肪组织，但吸收量大；细胞-支架复合体诱导组有大量脂肪类组织形成，油红染色显示组织有脂滴形成；细胞-支架复合体非诱导组和单纯支架组均未发现脂肪组织形成。结论采用组织下程技术将吸脂术获得脂肪来源细胞接种纤维凝胶支架，体内构建脂肪组织，具有可行性。     

Reversal of hyperglycemia in mice after subcutaneous transplantation of macroencapsulated islets

BACKGROUND: Macroencapsulated islets can reverse hyperglycemia in diabetic animals when transplanted i.p. or into the fat pad. The s.c. space is an attractive site for such transplantation because macrocapsules can be implanted with local anesthesia and be easily removed or reloaded with fresh islets. METHODS: Immunoprotective 20 microl ported devices were transplanted under the skin of Streptozocin-diabetic nude mice. Devices were loaded with 1200 rat islets in culture medium or in alginate. Empty devices were implanted for 2 weeks and then loaded with islets. Normal mice and mice with islets transplanted under the renal capsule or under the skin were used as controls. Seven weeks after transplantation, an intraperitoneal glucose tolerance test (IPGTT) was performed, followed by implant removal. RESULTS: Three weeks after transplantation, normal blood glucose levels were observed in all animals. Compared with those of normal controls, IPGTTs showed accelerated blood glucose clearance in mice transplanted with islets either within devices or beneath the kidney capsule. Fasted transplanted mice were hypoglycemic before glucose injection and 2 hr later. After removal of the implants, all recipient mice returned to hyperglycemia. Histological evaluation revealed viable islet cells and a network of close vascular structures outside the devices. CONCLUSIONS: Macroencapsulated islets transplanted into the s.c. space were able to survive and regulate blood glucose levels in mice. The observed differences in glucose metabolism between normal and transplanted mice may be attributed to the site of transplantation and to the use of rat islets, which have a different set point for glucose induced insulin release.     

稳定病理性瘢痕模型的建立

目的:通过比较两种病理性瘢痕动物模型——裸鼠瘢痕模型和兔耳瘢痕模型的稳定性并对制作方法进行改进,为病理性瘢痕研究中选择有效的动物模型提供依据.方法:分别采用改良保留表皮(带表皮人瘢痕组织移植1周后剪除移植组织上方的裸鼠皮肤)、去除表皮(去除瘢痕组织表皮后移植)的方法分别在皮下移植人增生性瘢痕和瘢痕疙瘩制作裸鼠瘢痕模型.分别采用去除软骨膜及软骨、保留软骨膜及软骨、去除软骨膜保留软骨3种术式制作兔耳瘢痕模型,观察各瘢痕模型的效果、模型稳定时间和病理学改变来评价模型质量.结果:裸鼠组可快速并稳定地复制增生性瘢痕及瘢痕疙瘩的病理生理特点,且改良保留表皮增生性瘢痕或瘢痕疙瘩移植组的稳定时间(15周和20周)及增生程度明显优于去除表皮的增生性瘢痕及瘢痕疙瘩移植组(模型稳定时间为8周和10周);兔耳组可观察到去除软骨膜保留软骨组瘢痕稳定时间(1 5周)及增生程度明显优于另外两组(模型稳定时间分别为9周和10周).结论:裸鼠瘢痕模型的制作以改良保留表皮移植术式为最佳,兔耳瘢痕模型的制作则以去除软骨膜保留软骨术式为最佳.     

携带Fas基因重组腺病毒治疗瘢痕疙瘩的体内实验研究

目的 研究携带人Fas基因的两种重组腺病毒对瘢痕疙瘩的体内治疗效果。方法 构建瘢痕疙瘩裸鼠模型,应用携带Fas基因的常规腺病毒Ad—Fas（T）和细菌内重组腺病毒Ad—Fas（B）注射及Fas单克隆抗体（FasMcab）辅助对植入裸鼠皮下的瘢痕疙瘩组织进行治疗。通过大体观察、常规病理及电镜观察检测瘢痕疙瘩组织的变化。结果 单纯使用腺病毒注射的瘢痕疙瘩组织块体积仅轻度缩小。前期注射Ad—Fas（B）或Ad—Fas（T）后,应用FasMcab作为后续治疗,瘢痕疙瘩组织块均明显缩小,HE染色证实瘢痕疙瘩组织结构遭到破坏,电镜观察发现细胞凋亡证据。结论 重组腺病毒Ad—Fas（B）及Ad—Fas（T）的瘢痕疙瘩基因治疗效果令人满意。为瘢痕疙瘩的治疗提供了新途径。     

基因重组腺相关病毒介导内皮抑素治疗祼鼠膀胱癌

目的 观察重组腺相关病毒(rAAV)-内皮抑素(Endostatin,ES)(简称rAAV-ES)基因治疗膀胱癌的效果.方法 T24细胞悬液接种于BALB/C裸鼠右侧肩胛区皮下,取已成瘤裸鼠随机分为3组:对照组、rAAV-多克隆位点(MCS)组、rAAV-ES组,每组5只,分别瘤内注射生理盐水、rAAV-MCS及rAAV-ES,4周后处死荷瘤裸鼠,取出移植瘤,常规苏木素-伊红(HE)染色、计算微血管密度(MVD);采集裸鼠血液标本酶联免疫吸附试验(ELISA)检测血清中ES浓度.结果 (1)瘤内注射rAAV-ES 9 d后,肿瘤生长明显抑制,治疗4周后:各组肿瘤体积分别是:rAAV-ES组(0.75±0.08)cm~3、rAAV-MCS组(1.27±0.13)cm~3和对照组(1.24±0.17)cm~3;组间比较差异有统计学意义(P＜0.05);(2)各组移植瘤MVD分别:rAAV-ES组(18.72±2.53)个/HP、rAAV-MCS组(52.38±6.46)个/HP和对照组(49.94±7.17)HP;rAAV-ES组显著低于对照组和rAAV-MCS组(P＜0.05).结论 rAAV-ES能抑制膀胱癌的血管生成和肿瘤的生长.     

Searching for the Favorable Donor Site for Fat Injection: In Vivo Study Using the Nude Mice Model

BACKGROUND: The use of suctioned fat grafts for correction of soft tissue defects is a widespread procedure in esthetic and reconstructive surgery. The main disadvantage of this simple and sensible procedure is the unpredictable absorption rate of the fat graft. A lot of research has been performed aiming for enhancement of the take of the fat grafts. OBJECTIVE: Our study was performed to find if there is any favorable donor site for fat harvesting. METHODS: This in vivo experiment using the nude mice model enables the study of the long-term survival of human fat in an animal model. The fat was harvested from three donor areas: the thigh, abdomen, and breast of a 48-year-old woman who came for an elective esthetic procedure. After centrifugation, 1 cc of fat was injected subcutaneously into the scalp of the nude mouse. There were 15 mice in each of the three groups, according to the selected donor sites. The animals were sacrificed 16 weeks after the procedure. The extracted fat was evaluated in terms of weight, volume, and six histologic parameters: integrity, vascularization, cyst formation, fibrosis, necrosis, and inflammation. RESULTS: This study could not find any statistically significant differences between the three investigated donor sites in the evaluated parameters. CONCLUSION: On the basis of this study, there is no favorable area for harvesting fat grafts. The donor site can be chosen according to the preference of the surgeon and the patient.     

轴状脂肪移植的实验和初步临床研究

目的通过与Coleman方法比较,探讨轴状脂肪移植方法的可行性。方法采用Coleman方法及轴状脂肪移植方法,分别于11例拟行头面部脂肪移植患者的下腹部抽取脂肪。取3～4周龄免疫缺陷裸鼠48只,雌雄不限,体重8.6～12.2 g;随机分为两组,每组24只。以背部皮下间隙为移植受区,实验组移植0.5 mL轴状脂肪移植方法获得的人体脂肪组织,对照组移植0.5 mL Coleman方法获得的人体脂肪组织。术后大体观察实验动物背部外观,于1、2、4、8周两组随机处死6只裸鼠,取出背部皮下移植脂肪进行大体、组织学及免疫组织化学染色观察,并于取出脂肪前后对裸鼠称重,计算移植脂肪重量。脂肪抽取术后即刻取脂肪组织进行葡萄糖转移实验和脂肪细胞活性测定。2010年5月-2011年10月,临床应用面部轴状脂肪移植治疗11例局部凹陷性畸形。结果术后实验组裸鼠背部隆起样外观基本维持,对照组2周时已趋于平坦。两组植入脂肪重量相似(P>0.05);除术后2周外,其余各时间点实验组残余脂肪重量均显著高于对照组(P<0.05)。组织学观察示实验组细胞形态保持良好,血管分布均匀;对照组细胞破坏再吸收显著,血管集中分布于移植组织边缘。术后实验组完整脂肪细胞计数均明显多于对照组,除术后2周外其余各时间点组间比较差异均有统计学意义(P<0.05)。实验组毛细血管计数均少于对照组,术后1、2周比较差异有统计学意义(P<0.05),但4、8周时差异无统计学意义(P>0.05)。轴状脂肪移植方法抽取的脂肪组织葡萄糖转移量为(1.462±0.080)mmol/L,显著高于Coleman方法抽取的脂肪组织(1.153±0.199)mmol/L(t=3.317,Ρ=0.021);且轴状脂肪移植方法抽取的脂肪组织细胞活性更佳。临床应用患者均获随访,随访时间2～9个月;随访期间受区未见明显萎缩及塌陷。结论与Coleman方法比较,轴状脂肪移植方法获得的脂肪组织可保持原结构和良好活性,移植术后能更早与受区建立血液循环,术后组织再吸收率低,形态维持时间长。     

荧光腹腔镜下小鼠胃癌可视化成像的实验研究

目的探讨荧光腹腔镜下小鼠胃癌可视化成像在鉴别肿瘤和正常组织中的价值。方法共20只3~4周龄,体重20 g裸鼠。2只裸鼠右后肢皮下注射0.2 ml高表达HER-2基因的NCI-N87胃癌细胞悬液用于观察背部成瘤情况,1周后当瘤体直径达1 cm左右进行下一步实验观察。将0.2 ml高表达HER-2基因的NCI-N87胃癌细胞悬液注射于12只裸鼠胃黏膜下,建立裸鼠胃癌模型。将量子点与HER-2的单抗结合制作量子点纳米荧光探针。将12只胃癌模型裸鼠分为A、B组,每组6只,A组裸鼠尾静脉注射量子点纳米荧光探针100μl与裸鼠胃癌模型进行体内结合,B组裸鼠尾静脉先注射HER-2单克隆抗体再注射量子点纳米荧光探针100μl,C组6只正常生长的裸鼠尾静脉直接注射量子点纳米荧光探针100μl。最后利用改装后的荧光腹腔镜分别以15、30 min和1、2、4、8、12、24、30 h时间点观察不同时期裸鼠体内胃癌组织及其他部位的荧光显像,标本送检。结果裸鼠种瘤成功,胃癌组织在荧光腹腔镜下完成可视化成像,荧光探针结合后稳定性良好,荧光强度在1 h后达到峰值,30 h后荧光消失。发光组织病理切片证实为胃癌组织,免疫组化证实该组织高表达HER-2蛋白。结论量子点纳米荧光探针性质稳定,荧光强度1 h达到峰值,然后逐渐衰减,荧光腹腔镜下可清晰看到高表达HER-2的NCI-N87胃恶性肿瘤的边界和周围组织及淋巴结的转移情况。     

脂肪组织来源干细胞提高游离脂肪移植存活率的研究

目的 探讨脂肪组织来源干细胞移植在体内促进游离移植脂肪组织的再血管化,提高移植脂肪组织存活率的可行性.方法 自人体吸脂术中脂质部分分离、培养AScs,行成脂、骨和软骨分化.将ASCs以DiI标记后与人体吸脂术获得的脂肪组织混合移植于18只裸鼠背部,裸鼠背部随机注入3组移植物:ASCs组(A组)、胰岛素组(B组)及培养基对照组(C组).术后6个月观察移植物情况,通过组织观察、HE染色和免疫组化进行分析.结果 抽脂术脂质部分能获取大量的ASCs,能分化成脂肪、成骨和软骨细胞.术后6个月3组移植脂肪的湿重分别为A组(165.97±5.51)mg,B组(93.42±5.12)mg,C组(67.64±5.09)lIIg.A组移植脂肪的存活率高于B、C组(P=0.000).纤维化程度的测定,"网格点计数"3组移植物的点个数分别为A组(152.2±9.8)个/10HF,B组(743.9±20.4)个/10HF,C组(892.2±16.5)个/10HF.A组移植脂肪的纤维化及坏死程度低于B、C组(P=0.000).免疫组化证实:ASCs散在分布于部分脂肪细胞及小叶间隔中,ASCs在体内能够部分转化为血管内皮细胞.结论 脂肪组织来源干细胞移植在体内可促进游离移植脂肪组织的再血管化,提高移植脂肪组织的存活率并改善了移植物的质地.AsCs辅助移植可能是一种较为理想的细胞疗法.     

结肠造口结肠癌原位移植模型的建立

目的建立一种新型的结肠癌原位移植模型—结肠造口结肠癌原位移植模型。方法对不同的BALB／C nu／nu裸鼠分别进行皮下结肠癌细胞接种和进行结肠造口,将皮下形成的肿瘤制成细胞悬液后种植在造口处,待肿瘤生长至5mm时使用氟尿嘧啶（5-Fu）腹腔注射。观察肿瘤生长、淋巴结转移及肿瘤的病理情况。结果10只造口成功的裸鼠全部生长出肿瘤,5只5-Fu处理组裸鼠的生存时间为（15.2±3.7）周,未处理组生存时间为（12.3±2.8）周,差异有统计学意义（P=0.001）。处理组中1例发现淋巴结转移,而未处理组有2例淋巴结转移（P=0.49）。病理检查：造口处生长的肿瘤皆为低分化癌,肠系膜淋巴结处为转移性肿瘤。结论结肠造口结肠癌原位移植模型是一种便于观察、可以反复取样且肿瘤生物学特性符合结肠癌的理想模型。