血液病自体脾组织移植的免疫结构：形态计量和免疫组化研究

应用形态计量法及免疫组化ＡＢＣ技术，检测１７例血液病自体脾组织移植后脾脏白髓的形态定量特征以及各区域淋巴细胞及其亚群分布。结果显示．血液病脾脏白髓内牌小体及生发中心增生、面积扩大．动脉周围淋巴鞘（ＰＡＬＳ）面积缩小；自体移植再生脾组织白髓脾小体及生发中心减少、面积缩小．红髓内淋巴细胞密度增加，以ＣＤ３、ＣＤ８增生为主。生发中心及边缘带ＣＤ２０明显减少．而ＰＡＬＳ内ＣＤ４减少、ＣＤ８增生。提示血液病自体脾组织移植后能重建脾脏的免疫结构．并能改善血液病脾脏免疫功能亢进及淋巴细胞的异常分布。     

Recurrent Ruptured Spleen

Regrowth of splenic tissue after splenectomy for trauma and splenectomy for idiopathic thrombocytopenia purpura have been reported. However, rupture of splenic tissue, either spontaneous or traumatic, that requires a second surgical intervention for hemoperitoneum caused by a ruptured splenic nodule or splenosis has rarely been reported. We report the case of a 43-year-old man in whom hemoperitoneum developed 25 years after he underwent an open splenectomy, after a motor vehicle accident, that required removal of a recurrent ruptured splenotic nodule.     

门静脉高压巨脾大部切除后残脾神经分布与定量研究

目的：观察门静脉高压巨脾大部切除后残脾神经纤维分布与密度变化,评估残脾保留的价值。 方法：选取门静脉高压脾肿大行脾大部切除并残脾腹后固定术患者13例,收集患者术后切取的巨脾组织,以及术后8年穿刺获取的残脾组织,另取外伤性脾组织13例为正常对照。采用免疫组化法检测脾神经肽Y（NPY）和神经丝蛋白200（NF 200）阳性神经纤维分布及密度。 结果：3组脾组织NPY和NF200阳性神经纤维的分布部位大致相同,但两者在巨脾组织中的密度明显较高。红髓部分的定量分析显示,巨脾组织NPY与NF200阳性神经纤维密度均明显高于残脾组织和正常脾组织（均P<0.05）,而两种阳性神经纤维密度在残脾组织与正常脾组织间差异无统计学意义（均P>0.05）。 结论：巨脾大部切除术后残脾神经纤维分布及含量与正常脾大致相同,提示解除高压环境后,残脾神经功能能逐渐恢复正常。     

The current and future aspects of liver cell transplantation

Our previous experiment presented that isolated adult rat hepatocytes survived and proliferated markedly and they reconfigured macroscopic hepatic tissue occupying more than 50% of the spleen 27 months after transplantation. On the basis of the results, we have emphasized that the hepatized spleen provides many fundamental and clinical interests on hepatology. The morphlogical studies of the recomposed hepatic tissue revealed almost normal architecture of the liver except the lack of bile duct system. Prolongation of the survival time was obtained in rats with hepatized spleen which underwent a portacaval shunt several months earlier compared with animals without hepatized spleen, when the rats were completely devascularized. That means that the hepatized spleen can act as an auxillary liver. For the purpose of applying this experimental model clinically, liver cell autotransplantation was performed using dog and monkey. These resulted in recomposed hepatic tissue in the monkey's spleen 15 months after transplantation, but no survival of the transplanted hepatocytes in dogs. In human, we have been developed a new hepatocyte isolation technique and we can obtain approximately 10(6) isolated human hepatocytes (viability 55-86%) from the partially resected liver with the hand-made multiperfusion system. Finally, future problems of clinical application of the intrasplenic hepatocyte transplant were mentioned especially about the idea of the liver cell bank.     

Perfusion and functional anatomy of the splenic remnant supplied by short gastric vessels

BACKGROUND: Surgeons have described many methods to preserve the injured spleen for the maintenance of host defence. The volume and the perfusion of remaining splenic tissue are important for better functional results. Imaging the functioning tissue with radionuclide provides information about the physiology of the concerning organ. METHODS: Thirty rats were separated into three groups: control, explorative laparotomy alone; partial splenectomy, upper part of the spleen supplied by short gastric vessels was preserved after partial resection; and devascularized spleen, the entire spleen was preserved after ligation of splenic artery. The size, functional anatomy, and perfusion status of splenic tissue were assessed by liver-spleen scintigraphy using radiolabeled heat denatured red blood cells. RESULTS: In the partial splenectomy group, splenic radioactivity count, spleen/liver ratio, and radionuclide uptake were mildly reduced, and found to be 87% (P = 0.012), 91% (P = 0.16), and 88% (P <0.001) of the normal spleen, respectively. The area of functional tissue in the upper splenic remnant was 51% (P <0.001) of the normal spleen. In the devascularized spleen group, the radioactivity count, the ratio, and the uptake were 38% (P <10(-6)), 36% (P <10(-6)), and 49% (P <10(-6)) of the normal spleen respectively. The area of functional tissue in the devascularized spleen was calculated as 47% (P <10(-6)) of the normal spleen. CONCLUSIONS: The collateral circulation is insufficient for proper function of the entire spleen after disconnection of the main arterial blood supply. The functional tissue is markedly impaired. On the other hand, the upper part of the spleen is remained well perfused via the short gastric vessels. The collateral circulation is satisfactory for this splenic tissue after reduction of its volume. That size splenic tissue seems to have satisfactory functional ability. Proper functional results mostly depend on the balance between the volume and blood supply of the remaining splenic tissue.     

Regeneration and phagocytic function of devascularized spleens

The phagocytic function of normal splenic tissue and of regenerated tissue following splenic artery ligation was investigated in rats using radiolabelled stannous fluoride colloid. Colloidal carbon was used to determine the histological location of phagocytosis within the spleen. Six months after ligation, the median weight of the devascularized spleens was 25% of that of spleens in control rats. Technetium stannous colloid clearance by devascularized spleens was reduced to 10% of normal and 25% when corrected for spleen weight. The colloidal carbon injected intravenously was observed primarily in the marginal zone in both normal and devascularized spleens. Histologically, devascularized spleens contained significantly less white pulp and marginal zone. The splenic tissue which regenerates following ligation does not have the phagocytic ability of normal splenic tissue. This may be due to the decreased regrowth of the lymphoid compartments of the spleen.     

YKL-40蛋白与门静脉高压脾脏肿大纤维化重塑进程的研究

目的：探讨YKL-40蛋白在人门静脉高压脾脏组织中的表达及临床意义。 方法：采用免疫组织化学法检测人正常脾脏组织14份及门静脉高压脾脏组织48份中YKL-40蛋白表达情况,Masson三色染色检测脾脏纤维化程度。 结果：门静脉高压患者脾脏YKL-40蛋白表达明显高于正常脾脏,两组YKL-40表达差异具有统计学意义（P 〈 0.05）,且其表达程度随Child-Pugh分级增高而增高（P 〈 0.05）。YKL-40蛋白表达与患者自由门静脉压力（FPP）呈正相关（R = 0.499,P 〈 0.01）。门静脉高压脾脏中纤维增生较正常脾脏增生明显（P 〈 0.05）,YKL-40蛋白表达与纤维化程度呈正相关（R = 0.857,P 〈 0.01）。 结论：YKL-40在门静脉高压脾脏中表达增高,且表达水平与FPP、Child-Pugh分级相关。YKL-40表达与脾脏纤维化程度成正相关,提示YKL-40可能参与门静脉高压脾脏纤维化重塑过程。YKL-40蛋白可能成为治疗门静脉高压脾大的重要生物治疗因子。     

Autologous splenic implantation in pigs. A comparison of two methods

Two methods of autologous splenic implantation, with sliced or with minced splenic tissue, were compared in six pigs. The two procedures were carried out simultaneously in each pig and the splenic remnants were surgically removed three months later. These remnants were histomorphometrically quantified by point counting. Statistical analysis showed the mincing procedure to be significantly superior, since 33% of the originally implanted tissue remained, in contrast to 21% of sliced, implanted tissue. At microscopy the tissue remaining after both procedures was in all cases indistinguishable from normal pig spleen. No necrotic splenic tissue was found. The authors conclude that minced tissue and omental pouch technique are preferable for autologous splenic implantation.     

Experimental study on function of regenerated splenic tissue after splenic autotransplantation

To prevent postsplenectomy overwhelming sepsis, splenic autotransplantation has been clinically attempted. However, function of regenerated splenic tissue after splenic autotransplantation has not been completely understood. Changes in weigh of regenerated splenic tissue, splenic blood flow, splenic immune responses and phagocytic function were studied for one year after splenic autotransplantation using Sprague-Dawley rats. At one year after autotransplantation, the weight of regenerated splenic tissue was increased to 80% of the originally implanted spleen and the blood flow was increased to 80% of the control spleen. The counts of lymphocytes and macrophages in the regenerated splenic tissue were significantly low at eight weeks after transplantation, however lymphocytes was increased to 58.8% and macrophages was increased to 29.5% of the control spleen at 16 weeks after transplantation. The blast formation of splenic lymphocytes was lower at the early stage after transplantation, thereafter, it was increased at the later time after transplantation. Microangiography of the regenerated spleen showed new capillaries around the implanted tissue 2 weeks after transplantation. These results suggested that the transplanted splenic tissue was regenerated to the similar structure to normal spleen and immunological function was recovered close to the normal splenic tissue.     

Alteration of proliferation and subtle changes of protein synthesis in autologous transplanted spleens.

In the case of massive splenic rupture, heterotopic autologous transplantation of splenic tissue into the omentum majus may be used to restore splenic function. Yet little is known about specific functions of the transplants compared to the normal spleen. The goal of this study was to get more information about immunologic functions and protein expression in splenic transplants. As an animal model we used the pig, whose splenic morphology and immunoarchitecture is similar to that of the human spleen. Histologic examination of transplants revealed structures that were comparable to normal spleens (consisting of red and white pulp and including germinal centers). Immunologic tests such as the hemolytic plaque assay and mitogen stimulation revealed that the number of plaque-forming cells was not changed significantly, but the stimulation index for T cells was drastically increased in the autotransplants. Electrophoresis and immunochemical methods showed differences in the protein patterns between both tissues. Several proteins were found to be produced only in the spleen, or were produced in much higher amounts in the spleen than in the splenic transplants. More information about these differences between spleen and splenic transplants is needed before we can recommend a general clinical application of autologous spleen transplantation.     

Autotransplantation of splenic tissue

Autotransplantation of splenic fragments has already been carried out in humans. The optimal size of the particles and amount of tissue required for this procedure has yet to be found. In normal young pigs and miniature piglets, autologous splenic tissue was transplanted into the greater omentum. The regenerated splenic mass, splenic blood flow, and histology were studied six months later. Implanting small splenic particles produced comparable results to implanting thin slices of splenic tissue. The mass of regenerated splenic tissue was only 3.1 g after implanting the whole spleen and 4.5 g after transplanting half of the spleen, which means 5.3% and 7.8% respectively of the weight of control spleens. The blood flow per gram in the regenerated splenic tissue was much lower than in the normal spleen. The blood flow in the whole of the splenic tissue is important for the clearance function of the spleen. Six months after transplanting the whole or half of the spleen, the blood flow to the regenerated splenic tissue was only 1% of that in the control minipigs. When half of the spleen was left in situ, as a model for a partial splenectomy, and the other half transplanted, the regenerated mass was only 3.4% of all splenic tissue and the blood flow 1.5% of the total splenic blood flow. In this model the regenerated splenic mass was independent of the size of the implants and the mass of implanted tissue. The extremely low blood flow indicates an inadequate clearance function and thus the protective function would probably be negligible.     

Ectopic spleen in the retroperitoneum. Case report

A 43-year-old man presented with a retroperitoneal mass, the histological examination of which showed well developed spleen parenchyma with a capsule and fibrous tissue in between normal looking splenic cells. Postoperative 99mTc sulphur colloid scans of his liver and spleen were normal. This is, to our knowledge, the first reported case of an ectopic spleen in the left retroperitoneal space.     

Torsion of Wandering Spleen in a Woman Presenting as Emergency

Wandering spleen is a very rare condition, it is characterized by the absence or weakness of one or more of the ligaments that hold the spleen in its normal position in the left hypochondrium. The disorder is not genetic in origin. Instead of ligaments, the spleen is attached by a stalk-like tissue supplied with blood vessels (vascular pedicle). If the pedicle is twisted in the course of the movement of the spleen, the blood supply may be interrupted or blocked (ischemia) to the point of severe damage to the blood vessels (infarction). Because there is little or nothing to hold it in place the spleen “wanders” in the lower abdomen or pelvis where it may be mistaken for an unidentified abdominal mass. “Acquired” wandering spleen may occur during adulthood due to injuries or other underlying conditions that may weaken the ligaments that hold the spleen in its normal position (e.g., connective tissue disease or pregnancy). We here report a case of 40-year-old woman presented to the surgical emergency department, with complaints of colicky abdominal pain of 8 h duration with increasing severity and bilious vomiting. On examination, the patient well built, nourished, conscious, and coherent with profuse sweating. Vital signs were normal excepting high pulse rate of 120/min with blood pressure of 100/60. On examination, she has a central abdominal distention shifting to different quadrants with colicky pain with vague tender firm mass. Abdominal ultrasonography showed ectopic spleen around the umbilicus with variable echos and whorled appearance of the splenic vessels. The patient was prepared for emergency laparotomy. At laparotomy, the spleen was found just below the umbilicus and it was dusky enlarged with infarction with four anticlockwise twists of the long splenic pedicle in the central quadrant of the abdomen. Splenectomy was carried out as there was clear evidence of splenic ischemia after detorsion of the spleen.     

Immune function of the upper splenic remnant supplied by short gastric vessels.

OBJECTIVE: To study the immune function of the upper third of the spleen supplied by short gastric vessels after two thirds partial splenectomy. DESIGN: Experimental study. SETTING: Teaching hospital, Turkey. MATERIAL: Sixty Wistar-albino rats, 20 in each group. INTERVENTIONS: Control = sham laparotomy; partial splenectomy = the upper third of the spleen supplied by short gastric vessels was preserved after two thirds partial splenectomy and dividing the main vascular supply; and total splenectomy. At the end of the sixth week postoperatively, antigenic stimulation was induced with an injection of pneumococcal suspension in 10 animals from each group. 0.5 ml of diluted Indian ink was injected into the aorta. MAIN OUTCOME MEASURES: Histological architecture of splenic tissue, and changes in the white pulp after antigenic stimulus. Bacteriological analysis with aerobic blood culture. Phagocytic activity as counted by Indian-ink-laden macrophages. The ability to produce antibodies as measured by serum IgM concentrations. RESULTS: Histological architecture of splenic tissue was normal. Germinal centres (p = 0.02), lymphoid follicles (p = 0.09), and their ratio (p = 0.0006) in the white pulp of the splenic remnant was significantly increased after antigenic stimulus compared with normal spleen. Significantly more animals without spleens developed bacteraemia (p = 0.02). Phagocytic activity of the upper splenic remnant was 89% that of normal spleen. Serum IgM concentrations without antigenic stimulus were 144, 138.2 (p = 0.6), and 86.2 (p < 0.001) mg/L; and with antigenic stimulus 263, 201.7 (p < 0.0001), and 98.1 (p < 0.0001) mg/L in groups 1, 2, and 3, respectively. The increase in serum IgM concentrations as a response to antigen was significant in the control (p < 0.0001) and in the partial splenectomy group (p < 0.0001), but not in the splenectomy group (p = 0.1). CONCLUSIONS: After reduction of its volume, the upper splenic remnant remained adequately supplied by the short gastric vessels. The upper part of the spleen preserved its normal histological architecture, had considerable phagocytic activity, possessed the ability to produce antibodies, and created a satisfactory immune response to antigenic stimulus. In rats, a considerable volume of functional and well perfused splenic tissue is preserved even after dividing the main vessels.     

Middle segment pancreatectomy for the treatment of benign and low malignant potential pancreatic tumours

Background: Conventional distal pancreatectomy is used for the removal of lesions located at the body and tail of the pancreas. Generally, the spleen is sacrificed. But for benign and low malignant potential tumours in the pancreatic neck and body, this incurs notable loss of normal pancreatic tissue and the unnecessary risk of diabetes mellitus as well as splenic loss. Methods: We report three cases of middle segment pancreatectomy in an effort to avoid the unnecessary loss of normal pancreatic tissue and the spleen. Two patients were males with a mucinous cystadenoma and non‐cystic mucinous cystadenocarcinoma. The other was a female with a serous cystadenoma. Middle segment pancreatectomy was performed. Two patients were reconstructed with double pancreaticojejunostomy and the third with a pancreaticogastrostomy. Results: No major complications were observed. Comnclusions: Middle segment pancreatectomy is a safe and effective procedure for the resection of benign and low malignant potential tumours in the pancreatic neck and body with preservation of normal pancreatic tissue and the spleen in selected patients. This is the first report of middle segment pancreatectomy for pancreatic tumour in China.     

磁性阿霉素白蛋白纳米粒在移植性肝癌模型中的磁靶向性

目的：观察磁性阿霉素白蛋白纳米粒在移植性肝癌模型中的磁靶向性，并观察磁性白蛋白纳米粒在各脏器中的分布特征。方法：建立大鼠移植性肝癌模型。大鼠正中开腹，胃十二指肠动脉插管固定。实验组，肝肿瘤区外加磁场，肝动脉注射磁性阿霉素白蛋白纳米粒(相当于阿霉素0．5mg／kg)，磁场应用30min，移去磁场后，动物立即处死。对照组肝肿瘤区不加磁场，肝动脉注射同样剂量的纳米粒后30min处死。动物处死后，立即取肿瘤组织、非磁区正常肝组织、心、肾、脾、肺、小肠和胃作了计数，肿瘤组织、肝组织送病理切片检查。结果：肝肿瘤区应用磁场30min后，磁区肿瘤组织的放射活性较非磁区肝组织的放射活性明显增加，磁区肿瘤组织的放射活性为非磁区正常肝组织的放射活性的8．7倍。对照组在没有磁场存在的情况下，肿瘤组织的放射活性为正常肝脏的2．8倍。实验组肺的放射活性较对照组明显降低。肾、心、脾、小肠和胃两组之间无明显差异。另外，实验组脾、肺和胃与肿瘤组织的放射活性之比较对照组大为降低。注入纳米粒800%以上分布于肝脏。结论：在磁场的作用下，磁性阿霉素白蛋白纳米粒在大鼠移植性肝肿瘤中的聚集明显增加。即使肝肿瘤区没有外加磁场，由于肿瘤组织和正常肝组织血管密度的差异，磁性阿霉素白蛋白纳米粒在肿瘤组织中的分布明显高于正常肝组织。实验组脾、肺、胃与肿瘤组织的放射活性比值大大低于对照组，说明磁场的存在使这些脏器的相对药物暴露明显降低。     

Optimal site and amount of splenic tissue for autotransplantation.

Clinical and basic studies have documented a high susceptibility to pneumococcal infection in asplenic humans and animals. It has been suggested that autotransplantation of splenic tissue might be a method of providing host resistance when total splenectomy is necessary. However, the effect of splenic autograft has remained controversial. This study was performed to evaluate the most effective site and amount of splenic autograft using rats. Rats were divided into five groups for the purpose of determining the site of splenic autotransplantation: splenectomy, sham operation, implantation into the omental pouch, intraperitoneal implantation, and intramuscular implantation. For determining the amount for autotransplantation, the rats were divided into seven groups: splenectomy, sham operation, and implantations of 25, 50, 100, 200, or 300 mg of splenic tissue. All animals were challenged with Streptococcus pneumoniae type 6, 16 weeks after surgery. Howell-Jolly bodies appeared postsplenectomy, but disappeared in the implanted rats 16 weeks after the operation. Histologically, the implanted tissue was indistinguishable from that of a normal spleen. Pneumococcal clearance from the bloodstream and survival rate were significantly higher in rats implanted in the omental pouch as compared with splenectomized rats. Intraperitoneal and intramuscular implanted rats did not show a significant difference from the splenectomized rats. More than 50% of splenic tissue for autograft showed a significant increase in pneumococcal clearance and survival rate as compared with that of splenectomized rats. It was suggested that the most effective site of autotransplantation is the omental pouch and approximately 50% of the whole spleen would be necessary for prevention from sepsis.     

Immune cell subpopulations in regenerated splenic tissue in rats.

BACKGROUND: Asplenic patients have an increased risk of infections. Operations such as autotransplantation or splenic artery ligation have been suggested to ensure retention of functional splenic tissue after splenectomy, but their protective value is unclear. Immune responses, such as production of antibody, remain impaired in humans and animals even when such tissue is present, and phagocytosis is less efficient than by normal spleen tissue. In the present study the cellular composition of regenerated tissue is determined. METHODS: Splenic tissue was obtained from rats 6-9 months after splenic autotransplantation, splenic artery ligation or sham operation. The lymphocyte and macrophage subpopulations were labelled using a panel of monoclonal antibodies and analysed by flow cytometry. RESULTS: Both the total number of cells and the number of cells per gram of tissue were significantly reduced. There was a substantial reduction in the percentage of some of the cells examined (CD4+ and CD8+ T lymphocytes subsets), but not all (B lymphocytes, ED1+ and ED2+ macrophages, OX2+ and OX6+ cells). CONCLUSIONS: The reduction in the T lymphocyte subsets in regenerated splenic tissue compared with the normal spleen might explain the immunological dysfunction which persists after splenic autotransplantation. The reduction in the number of macrophages may be responsible for the alteration in phagocytic efficiency of regenerated splenic tissue.     

A study of liver regeneration using fetal rat liver tissue transplanted into the spleen

The liver morphology of fetal hepatic tissue transplanted into an ectopic location was investigated over one year period. Fetal liver fragments prepared from a maternal rat on the 18th or 19th day of pregnancy were injected into the splenic parenchyma of syngeneic rats using a 21 gauge needle. Histologically, the fetal liver did not essentially show any apparent lobular architecture or cord structure. The transplanted fetal hepatic tissues survived and formed hepatic cords in the spleen instead of undergoing degeneration and necrosis. Three characteristic features became complete during the 4 weeks following transplantation, namely; clumps of hepatocytes with obvious hepatic cords and sinusoids, markedly proliferating bile ducts and proliferating individual hepatocytes. Macroscopic nodules of the hepatocytes on the spleen were seen at about 6 months after transplantation. When the differentiation of the transplanted fetal hepatic tissue was compared with the development of a normal neonatal liver after birth, it was delayed by only about one week, while there was no proliferation of bile ducts in the normal neonatal liver. This experimental model provides a useful system for investigating liver regeneration and the mechanism of cell growth.     

A study of liver regeneration using fetal rat liver tissue transplanted into the spleen

The liver morphology of fetal hepatic tissue transplanted into an ectopic location was investigated over one year period. Fetal liver fragments prepared from a maternal rat on the 18th or 19th day of pregnancy were injected into the splenic parenchyma of syngeneic rats using a 21 gauge needle. Histologically, the fetal liver did not essentially show any apparent lobular architecture or cord structure. The transplanted fetal hepatic tissues survived and formed hepatic cords in the spleen instead of undergoing degeneration and necrosis. Three characteristic features became complete during the 4 weeks following transplantation, namely; clumps of hepatocytes with obvious hepatic cords and sinusoids, markedly proliferating bile ducts and proliferating individual hepatocytes. Macroscopic nodules of the hepatocytes on the spleen were seen at about 6 months after transplantation. When the differentiation of the transplanted fetal hepatic tissue was compared with the development of a normal neonatal liver after birth, it was delayed by only about one week, while there was no proliferation of bile ducts in the normal neonatal liver. This experimental model provides a useful system for investigating liver regeneration and the mechanism of cell growth.