自体移植脾组织神经肽Y阳性神经纤维再生的实验研究

目的：研究自休脾组织移植术后不同时相神经肽Y阳性（NPY^ ）神经纤维的再生规律。方法：健康Wistar大鼠56只，雌雄不限，体重100-120g，随机分为实验组及假手术组，术后7，14，30，60，90，120，180d取两组脾组织，行免疫组化抗NPY抗体染色及图像分析定量测定。结果：术后30d内，自体移植脾组织无NPY^ 神经纤维；术后60d，NPY^ 神经纤维出现于移植脾组织周边区域；术后90d，NPY^ 神经纤维向移植脾组织中央实质区伸展；术后120-180d，移植脾组织内NPY^ 神经纤维分布及密切均接近正常。结论：自体脾组织移植后可实现NPY^ 神经纤维再生；再生神经纤维来源于移植脾组织周围大网膜。     

自体脾组织移植后病理学变化的实验研究

目的　研究自体脾组织移植后不同时相点病理形态学变化规律。　方法　健康Wistar大鼠 5 6只 ,雌雄不限 ,体重 10 0～ 12 0 g ,随机分为 7组 ,每组 8只中又设脾切除自体组织大网膜内脾移植组 5只 ,假手术组 3只 ,分别于术后 7、14、3 0、60、90、12 0、180d取脾组织 ,光镜、透射电镜观察。　结果　移植脾组织之重量在术后 7d最轻 ,为 0 .0 72 g ,其后逐渐增加 ,180d时为 0 .5 11g ,各时相点之间有明显差异 (P <0 .0 5 )。病理检查提示移植脾组织经历急性期、缓解期、修复期三个病理时期 ,逐步恢复并接近正常的组织结构。　结论　自体脾组织大网膜内移植术是简便有效的脾移植方法。自体移植脾组织再生过程可分为 :急性期、缓解期与修复期     

自体脾泥移植治疗创伤性脾破裂

目的：探讨自体脾泥移植在创伤性脾破裂全脾切除术中的疗效。方法：对11例创伤性脾破裂患者行全脾切除及自体脾泥组织大网膜内移植术,观察其临床疗效。结果：本组1例因合并严重肺损伤术后死于呼吸衰竭,1例继发腹腔感染失败,9例治愈;术后3个月复查B超或CT有移植脾存活,血清IgM,IgG恢复正常范围,无脾切除后暴发性感染(OPSI)发生。结论：对创伤性脾破裂行全脾切除患者,自体脾泥大网膜移植是一种新的、安全、有效的保脾手术方式。     

带蒂大网膜修复严重输尿管损伤及其机制的实验研究

目的 探讨带蒂大网膜修复严重输尿管损伤的作用机制. 方法 20只健康成年杂种犬随机分为实验组及对照组,每组10只,均建立严重输尿管损伤动物模型,端端吻合输尿管后实验组采用带蒂大网膜包裹损伤输尿管,对照组未采用大网膜包裹.术后不定期观察有无尿瘘及输尿管坏死.术后12周观察术侧输尿管损伤愈合情况,取吻合口及周围组织行病理学检查,镜下观察血管再生情况,免疫组化检测血管内皮生长因子(VEGF)及其受体KDR的表达. 结果 术后实验组无尿瘘,对照组2只因尿瘘反复腹腔感染死亡.术后12周实验组输尿管吻合口处黏膜及平滑肌层完全再生,血管再生现象明显,VEGF及KDR表达阳性细胞密度分别为(12.65±0.02)%和(10.23±0.03)%.对照组吻合口愈合不良并狭窄,思侧肾严重积液(脓),无明显血管再生,VEGF及KDR表达阳性细胞密度仅为(1.54±0.03)%、(2.65±0.04)%,明显低于实验组(均P<0.05). 结论 带蒂大网膜有促进严重输尿管损伤修复作用,可能通过VEGF及KDR表达升高促进血管再生而实现.     

带蒂大网膜对严重损伤输尿管的修复作用

目的 探讨带蒂大网膜对严重输尿管损伤的修复作用及其机制.方法 随机将20条犬均分为实验组及对照组,建立严重输尿管损伤动物模型,实验组采用带蒂大网膜包裹损伤输尿管,对照组未采用大网膜包裹.术后观察尿瘘及输尿管坏死情况,术后12周再手术观察输尿管损伤愈合及吻合口血管再生情况,免疫组织化学染色检测血管内皮生长因子(VEGF)及其受体KDR的表达.结果 实验组均无尿瘘,对照组2例因尿瘘反复腹腔感染而死亡.术后12周实验组输尿管吻合口处黏膜及平滑肌层再生,血管再生现象明显,VEGF及KDR表达升高.对照组吻合口处愈合不良或瘢痕愈合,血管再生不明显,VEGF及KDR阴性或弱表达.结论 带蒂大网膜有促进严重输尿管损伤修复的作用,可能通过VEGF及KDR的表达升高促进血管再生而实现.     

自体移植脾巨噬细胞C3b、Fc受体及蛋白质表达的实验研究

目的 探讨自体脾组织移植后的功能状况。方法 采用小鼠进行自体脾组织网膜内移植，术后6个月切取移植脾组织，检测巨噬细胞的Fc、C3b受体及蛋白表达。结果 自体移植脾巨噬细胞Fc受体的含量与原位脾相近，C3b受体的功能正常；蛋白质的表达与原位睥相同。结论 大网膜内自体移植脾组织的功能在细胞水平是正常的，移植脾组织具有原位脾的功能。     

腹膜后自体脾移植治疗外伤性脾破裂的前瞻性研究

目的 探讨腹膜后自体脾移植在严重脾外伤保脾手术中应用的可行性及价值。方法 采用前瞻性病例对照研究,将66例严重脾外伤病人分为腹膜后自体脾移植组例、大网膜自体脾移植组,其中腹膜后自体脾移植组32例,大网膜自体脾移植组34例。观察术后一般情况,术后1天～ 12个月血常规,血IgA、IgM、IgG、C3、Tuftsin水平变化。结果 腹膜后自体脾移植组手术时间短于大网膜自体脾移植组（P<0.05）,术后其它一般情况变化、术后1天～ 12个月血IgA、IgM、IgG、C3、Tuftsin水平两组无统计学差异（P> 0.05）。结论 腹膜后自体带蒂脾移植术能够保留脾脏的部分免疫功能,且手术操作简便,在临床上推广应用是可行的。     

转染VEGF基因提高游离自体颗粒脂肪移植物存活率的实验研究

目的 研究转染人血管内皮生长因子基因(VEGF)对大鼠游离的自体脂肪移植后移植物存活的影响.方法 SD大鼠48只,分为3组,每组16只.于自体游离颗粒脂肪移植时分别注入脂质体包裹的重组VEGF质粒(目的 基因组)和空白质粒(空白质粒组)以及生理盐水(生理盐水组).术后定期计算植入物后前重量比,并行HE染色观察组织病理改变,免疫组化法检测组织的VEGF表达水平及微血管密度.结果 目的 基因组在移植物重量改变方面显著小于空白质粒组及生理盐水组(P＜0.05),其VEGF表达及微血管密度均显著高于其他两组(P＜0.01),空白质粒组及生理盐水组差异无统计学意义.结论 脂质体包裹的VEGF基因质粒能够在脂肪组织中表达VEGF,诱导新血管的形成,减少移植组织的吸收.     

脾损伤自体脾组织移植的临床应用

目的：探讨自体脾组织移植在临床中的应用。方法：总结32例脾外伤行全脾切除自体脾组织移植手术，其中采用大网膜囊内移植18例，去粘膜游离空肠段内移植12例，腹直肌鞘内移植2例。结果：术后随访均显示脾功能满意，尤以去粘膜游离空肠段内移植效果最好。结论：自体脾组织移植可作为严重脾外伤、全脾切除术后保留脾功能的一个重要有效手段，移植脾的功能恢复与血供有密切的关系。     

不同量自体脾组织移植抗肺炎球菌感染的研究

采用大鼠进行不同量脾组织网膜内移植，术后６月检测有关抗感染指标，结果显示气管内感染肺炎球菌后，４０％，６０％，８０％脾移植组和假手术组的存活率明显高于脾切除组，血中肺炎球菌廓清率在脾切除组中明显下降，而在各移植组则接近正常，结果表明，自体脾组织移植有一定上的抗感染能力，脾组织移植量以４０％～６０％为宜。     

Hematological, hemorheological, immunological, and morphological studies of spleen autotransplantation in mice: preliminary results

Using a spleen autotransplantation model, we conducted hematological, hemorheological, immunological, and morphological studies in mice 6 weeks after splenectomy. Sixty male and female A/J inbred mice were equally divided into 3 groups: 1) SE group, splenectomy was performed; 2) AU group, spleen chips were autotransplanted into the omentum without vascular anastomosis following splenectomy; and 3) C group (controls), no intervention in these mice. At postoperative week 6, the following studies were performed: 1) measurement of hematological parameters; 2) hemorheological studies, including relative cell transit time (RCTT) and fibrinogen levels; and 3) activity of peripheral phagocytes, measured by zymozan-induced chemiluminescence, which was calculated in stimulation index values (SI). In addition, histological investigations of autotransplants were conducted. Erythrocyte mean cell volume and platelet counts, RCTT, fibrinogen levels, and activity of phagocytes were significantly higher in the SE group, compared to those in the C group. In the AU group, these parameters were similar to those in the C group. Morphologically, the transplanted spleen showed normal histology. These data indicate that the transplanted spleens restored their function. We conclude that spleen autotransplantation reserves the normal morphology of spleen and restores most of the spleen's hematological, hemorheological, and immunological functions. Both SI index and erythrocyte deformability can be an informative detection of decreasing splenic function. These data suggest that spleen autotransplantation may provide a useful tool to prevent complications following splenectomy in a clinical setting.     

Regeneration of splenic remnants after partial splenectomy in rats

Splenic regeneration in the rat was measured after removal of 25, 50, or 75% of the spleen, 50% of the spleen with autotransplantation of the excised portion, and splenectomy with autotransplantation of 50% of the spleen. Splenic growth in rats undergoing sham splenectomies served as a control. Splenic mass at 6 weeks and 4 months after surgery was directly related to the remnant size. “Normalized” spleen weights (measured as grams of splenic tissue per 100 grams of rat weight) after 25, 50, and 75% splenectomy were 57, 41, and 38% of controls at 6 weeks, and 77, 71, and 44% of controls at 4 months. All differences were significant at P < 0.03 except those between 50 and 75% splenectomy at 6 weeks, and between 25 and 50% splenectomy at 4 months. A comparison of autotransplanted splenic mass after total splenectomy with that after 50% splenectomy (0.042 ± 0.005 and 0.025 ± 0.004, respectively, at 6 weeks) demonstrated that an intact subtotal spleen inhibited significantly regeneration of the autotransplanted spleen. The effect of autotransplanted splenic tissue on regeneration of a splenic remnant was little to none at 4 months.     

Assessment of post-splenectomy residual splenic function. Splenic autotransplants

Splenectomy increases the risk of fulminant sepsis. The present study assesses residual splenic function in patients splenectomized due to traumatic rupture of the spleen; and six cases with splenic autotransplants. Splenic tissue was observed in only 48% of the splenectomized patients and 100% of the autotransplant cases. The two most reliable analytical parameters to assess the presence of functional splenic tissue, were the absence of Howell-Jolly bodies and normal IgM blood levels. In cases where total splenectomy is indicated, it has proved useful to perform autotransplantation of splenic tissue at omentum major level.     

Laparoscopic splenic autotransplantation

Since 1990, we have performed splenic autotransplantation in more than 100 patients to treat splenic trauma, portal hypertension, myeloid metaplasia due to myelofibrosis, chronic lymphocytic leukemia, and Gaucher disease. The aim of this present study was to present splenic autotransplantation performed by laparoscopic means. A 33-year-old woman with severe splenic pain due to ischemia caused by multiple focal thromboses of splenic arterial branches was successfully treated by laparoscopic splenectomy and splenic tissue autotransplantation. The spleen was removed and cut in 20 fragments that were sutured to the greater omentum. This procedure was safely conducted with minor bleeding and no technical difficulties or complications. The postoperative follow-up of 12 months has been uneventful; the patient's pain disappeared on the first postoperative day. Hematologic, immunologic, tomographic, and scintigraphic examinations confirmed the functions of the splenic autotransplants. It is feasible and safe to perform splenic autotransplants by laparoscopic means.     

Splenic tissue autotransplantation in rabbits: no restoration of host defense

BACKGROUND: The loss of spleen may increase the incidence of overwhelming sepsis. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the effectiveness of regenerated splenic tissue in preventing infection. This study explored the effectiveness of splenic tissue autotransplantation in restoring host defense. MATERIALS AND METHODS: Rabbits were divided into three groups: splenic autotransplantation, sham operation, and total splenectomy. Histomorphology, T-lymphocyte count, serum lysozyme levels, hemolysin titers, and pneumococcal clearance were observed as read-out parameters over 24 weeks. RESULTS: Histological study showed that the white pulp was poorly developed and central arterioles were missing in the regenerated splenic tissue of the autotransplanted rabbits. The weight of regenerated spleens recovered 6 months later in the splenic autotransplantation group was 11% of that in the sham operation group and was significantly less than the weight at implantation. There was no significant difference in the number of T lymphocytes or level of serum lysozyme between the three groups. A poor antibody response by the rabbits in the splenic autotransplantation and total splenectomy groups was noted after the primary intravenous administration of sheep red blood cells compared to those of sham operation group. After the challenge with type 3 pneumococci intravenously, pneumococcal clearance from the bloodstream in the splenic autotransplantation group did not differ significantly from that in the total splenectomy group, but was markedly delayed compared with that in the sham operation group. CONCLUSIONS: The low quantity and poor quality of the regenerated splenic tissue contribute to the inferior immunoprotective ability of animals autotransplanted with one-third of the original spleen. This suggests that the regenerated spleen cannot compensate for the immunological function of the original one, especially host resistance to infection.     

Histologic study of experimental spleen transplant in rats

BACKGROUND: The objective of this paper was to demonstrate that the grafts of cervical splenic transplantation on rats using our experimental model present a normal histological appearance. METHODS: Isogenic consanguineous Lewis rats 12 weeks old and weighing 250 gr. were used. Histological findings of a group of 25 cervical splenic grafts transplanted by means of splinting vascular venous microanastomoses and a group of 25 splenic grafts autotransplanted in the omentum were compared with a control group. The specimens were assigned according to a score of 0 to 4, following Moore's histological criteria. RESULTS: All grafts in transplanted and autotransplanted groups had a score of 3 or 4. Then, all splenic grafts from the transplanted group had histological findings very similar to a normal spleen. In the autotransplantation group, the percentage of grafts with a score 3 (60%) was superior to the transplantation group (46%). However, the transplantation group presented a percentage of score 4 (54%), superior to the autotransplantation group (40%). CONCLUSIONS: In our study all grafts from the cervical spleen transplantation group had histological findings very similar to a normal spleen. The percentage of spleens with histological normality in the transplantation group was superior to the autotransplantation group. However, there was no statistical significance.     

Immunologic function against infection in splenic autotransplanted mice

The increasing recognition of the danger of overwhelming postsplenectomy infection (OPSI) has led surgeons to attempt to maintain splenic function after spleen injury. One technique they use when splenorrhapy or partial splenectomy are not feasible is the deliberate autotransplantation of splenic tissue. But the amount of splenic tissue necessary to prevent OPSI remains controversial, and opinions differ about the importance of the location and size of the splenic fragments implanted. The mice were divided into five groups, I. splenectomy, II. splenectomy +30% of the spleen implanted intraperitoneal site, III. splenectomy +50% implanted intraperitoneally, IV. splenectomy +50% implanted subcutaneously and V. Sham operation. This study assessed the blood flow of the splenic tissue, increasing weight of splenic mass, histology, the serum level of the immunoglobulins (IgG, IgA, and IgM), pneumococcal antibody titers after vaccination, and survival after intravenous pneumococcal challenge. This study demonstrated that intraperitoneal transplantation showed better regeneration and afforded better protection from OPSI than subcutaneous transplantation. And 30 to 50 percent of the whole splenic tissue mass protected against experimental pneumococcal sepsis. The splenic autotransplants developed in volume and blood supply after 8 weeks, and immunologic function against infection recovered at the same time.