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[Laparoscopic surgery of the spleen in children].

Laparoscopic operations of the spleen, incl. splenectomy, are increasingly accepted worldwide. The most frequent indications for this type of surgical treatment are haematological diseases (some types of haemolytic anaemia, thrombocytopenia, myeloproliferative syndrome). Another indication for surgery is hypersplenism from other causes such as vascular anomalies or isolated thrombosis of the lienal vein. The reason for urgent or planned laparoscopy may be also traumatic damage of the spleen (haemorrhage, pseudocysts). In the submitted work the authors present their experience with 21 patients where a laparoscopic operation of the spleen was performed. Of 21 children four were laparoscopically operated on account of a mesothelial cyst or posttraumatic pseudocyst of the spleen (ratio 1:3). In another 15 children laparoscopoc splenectomy was performed for haematological reasons (11 x spherocytosis, 3 x idiopathic thrombocytopenic purpura, 1 x autoimmune haemolytic anaemia). Twice the operation was indicated on account of isolated thrombosis of the lienal vein. In three instances a concurrently, also laparoscopic cholecystectomy was performed and in two liver biopsy. On operation 3-5 troacars were used and the patient was placed in a supine position. In no instance preoperative embolization of the lienal arteries was performed. On treatment of cystic structures of the spleen the authors made in three instances total resection of a cyst--in the upper pole of the spleen, in case of a central localization, only 2/3 resection and fixation of the omentum. In resections recently coagulation and Argon spraying was used. The spleen was extracted by morcellation in a plastic bag or via a 5 cm left-sided subcostal incision (2x). The median size of the spleen was 14.4 cm (10-20 cm), of cystic formations 7.25 cm (6-8 cm). The mean period of operation in splenectomy was 146 minutes. In two patients a blood transfusion was necessary (1x splenectomy, 1x resection of the lower third of the spleen). As to complications the authors recorded once major haemorrhage after splenectomy--resolved by the use of Endo GIA stapler. Conversion was not necessary in any of the operations. The mean period of hospitalization was 6 days.

Adolescent↗

[Traumatic rupture of the spleen].

In the work the authors present their own experience in the treatment of spleen traumatic rupture in the period of 10 years (1978-1987). In that period in 13 patients spleen were removed because of traumatic rupture. Spleen iatrogen lesions, spontaneous spleen ruptures and spleen-removing caused by other indications were not considered. The 6 male and 7 female patients were operated on the 6 year old child was the youngest and 79 year old man was the oldest. Spleen traumatic rupture is mostly caused by falls, accidents (traffic) and then by blows and fights. 4 patients had isolated spleen injuries and 9 spleen injuries together with neighbouring organs injuries. Some postoperative complications were noticed as: slowly wound healing (1), ileus (1), bronchopneumonia (2) and hematothorax (4). There was no mortality The authors consider each spleen traumatic rupture should not be removed especially in children and others with light injuries.

Adolescent↗

Regulation of apoptosis in vitro in mature murine spleen T cells.

Previously it has been shown that thymocytes undergo apoptosis, a form of programmed cell death, in response to glucocorticoids. This classic form of apoptosis is prevented by inhibition of protein synthesis. The current paper demonstrates that mature T cells also undergo apoptosis, but that the regulation of apoptosis in spleen T cells differs from that of thymocytes. Mature mouse spleen T cells were shown to die by apoptosis, not necrosis, when cultured without an added stimulus. Assays for apoptosis included internucleosomal DNA cleavage by gel electrophoresis, percent fragmentation of DNA by the diphenylamine method, and percent of cells with hypodiploid DNA by flow cytometry. The percent of apoptotic cells was 2% in fresh spleen T cells, and increased at least until 16 h, when 21% were apoptotic. Dexamethasone caused apoptosis in both thymus and spleen T cells, but only thymocytes showed a requirement for protein synthesis in dexamethasone-induced death. Cycloheximide increased apoptosis in spleen T cells, indicating that apoptosis was controlled by newly synthesized protective proteins. Spontaneous apoptosis was decreased in spleen T cells by protein kinase C activation, and was increased by H7 and staurosporine, which inhibits protein kinases, in contrast with the behavior of thymocytes. The protein kinase A/G inhibitor HA1004 also decreased spleen T cell apoptosis. The contrasting effects of cycloheximide on thymocytes and spleen T cells occurred over the same concentration range, and the same was true for PMA. The dexamethasone dose-response curves were similar, except that a greater proportion of spleen T cells were dexamethasone-resistant. These data support the hypothesis that the apoptosis program in T cells undergoes a transition during their maturation, such that apoptosis in mature T cells is regulated more like that of mature B cells than that of thymocytes.

Animals↗

The wandering spleen: CT findings and possible pitfalls in diagnosis.

AIM: To report the CT features of wandering spleen, a rare condition which can be incidentally detected as an abdominal or pelvic mass or can present with torsion, causing an acute abdomen. MATERIALS AND METHODS: The CT studies of seven patients, two children and five adults, with wandering spleen were reviewed. CT was performed urgently in three patients for acute abdomen, and electively in four. RESULTS: CT findings of wandering spleen included absence of the spleen in its normal position and a mass located elsewhere in the abdomen or pelvis, i.e. an ectopic spleen, enhancing homogeneously in four cases and failing partially or completely to enhance in the other three, indicating infarction. A "whirl" appearance representing the twisted splenic pedicle was seen in the three cases with torsion. Urgent splenectomy confirmed infarction secondary to torsion. CONCLUSION: The possible diagnosis of wandering spleen should be kept in mind when CT shows the spleen to be absent from its usual position and a mass is found elsewhere in the abdomen or pelvis. When, in addition, a "whirl" or partial or no enhancement of this mass are seen in a case presenting with acute abdomen, torsion of a wandering spleen is a likely diagnosis.

Abdomen, Acute↗

Pneumococcal clearance function of the intact autotransplanted spleen.

OBJECTIVE: To determine if the venous drainage of the spleen into the portal circulation is essential for its ability to protect against encapsulated bacterial challenge. DESIGN AND INTERVENTIONS: Three groups of dogs were randomly assigned to undergo either sham laparotomy, splenectomy, or splenectomy with autotransplantation of the intact spleen into the pelvis and formation of vascular anastomoses to the iliac vessels. Two weeks post-operatively, the dogs received a sublethal intravenous injection of Streptococcus pneumoniae type 25. OUTCOME MEASURES: Bacterial clearances and inflammatory damage to the liver. Bacterial clearance of the autotransplanted spleen should be no different from that of a sham-operation spleen and significantly different from that of a splenectomized animal. In addition, immunologic function of the autotransplanted spleen should not differ from that of a sham-operation spleen in the degree of inflammatory damage to the liver. RESULTS: No differences in bacterial clearance function were found between the animals that had undergone sham laparotomy or splenic autotransplantation. However, bacterial clearance in the splenectomized animals was significantly impaired. Histologic examination of the liver 2 weeks after the bacterial challenge revealed high-grade inflammatory damage to the livers of splenectomized dogs, intermediate liver damage in dogs that underwent autotransplantation, and essentially no damage in dogs that underwent sham laparotomy. Autotransplanted spleens were essentially nonreactive, lacking actively proliferating germinal centers, whereas splenic tissue from sham-operation animals showed reactivity. CONCLUSION: Although bacterial clearance function is unchanged in autotransplanted spleens, this method still does not fully protect the liver from inflammatory damage.

Analysis of Variance↗

Hematopoiesis in the human spleen.

To assess the capacity of the human spleen to function as a hematopoietic organ, spleen mononuclear cells from patients undergoing splenectomy were cultured to assay CFU-E, BFU-E, and CFU-C. Although the peripheral blood from cases of hereditary spherocytosis, Gaucher disease, Thalassemia major, and idiopathic thrombocytopenic purpura yielded 3-20 BFU-E/5 X 10(5) cells plated and 1-7 CFU-C/5 X 10(5), suspensions of spleen cells failed to grow colonies. Histologically these spleens did not show extramedullary hematopoiesis. In contrast, peripheral blood from a patient with osteopetrosis (marble bone disease) yielded 130 BFU-E/5 X 10(5), 40 CFU-E/5 X 10(5), and 9 CFU-C/5 X 10(5). The spleen had extensive extramedullary hematopoiesis on microscopy and grew 180 BFU-E/10(5), 210 CFU-E/10(5), and 25 CFU-C/10(5). We conclude that spleens from patients without extramedullary hematopoiesis do not contain committed hematopoietic progenitors in spite of normal precursors in the blood. In osteopetrosis the spleen contained stem cells in a concentration comparable to bone marrow, and the embryonic role of the spleen in blood formation appeared to continue in postnatal life.

Colony-Forming Units Assay↗

CD5 positive immunoregulatory B cells in spleen populations from multiple myeloma patients.

CD19+CD5+ lymphocytes constitute a minority of peripheral blood B cells. In view of the importance of these cells in the pathogenesis of the immunoregulation of myeloma, their incidence in another lymphoid organ was determined. CD5+ B cells were studied in 9 spleens from patients with multiple myeloma and in 10 spleens from normal individuals removed secondary to trauma. The total number of CD19+ B cells were increased in myeloma spleens (44.4% +/- 12.6%) as compared to normal spleens (20.4% +/- 7.4%). Likewise, the percentage of CD19 cells which co-expressed CD5 were increased in myeloma (25.3% +/- 12.4%) versus normal (4.4% +/- 2.3%) spleen. CD5+ B cells isolated from myeloma spleens, but not normal spleens, inhibit production of immunoglobulin in a pokeweed mitogen driven assay. Thus the spleen appears to be an important source of immunoregulatory B cells in multiple myeloma.

Antigens, Differentiation↗

An electron microscopic study of the spleen of the rat in an acute myelogenous leukemia.

The spleens of rats were studied by light and electron microscopy during the course of an acute myelogenous leukemia, with special reference to infiltration of leukemic myeloblasts in the spleens and to the correlation of leukemic cell infiltration with splenic hematopoiesis and splenomegaly. Leukemic myeloblasts infiltrated the cordal space of the red pulp. Many of them appeared in groups. Even in spleens which were heavily infiltrated, leukemic myeloblasts did not penetrate the while pulp. Massive infiltration and proliferation of the leukemic myeloblasts in the red pulp resulted in splenomegaly. The spleen increased its hematopoietic activity, while the medullary hematopoiesis diminished due to the invasion of leukemic myeloblasts in the bone marrow. Compensatory splenic hematopoiesis occurred in most of the leukemic spleens, but diminished in spleens which were very heavily infiltrated with leukemic myeloblasts. Thus, the degree of splenomegaly and splenic hematopoiesis did not necessarily correspond to the percentage of leukemic myeloblasts in the bone marrow, but rather related to the number of leukemic myeloblasts present in the spleen. A possible role for the splenic sinus walls in promoting compensatory hematopoiesis in the spleen is discussed. A consistent association of type "C" virus particles with leukemic myeloblasts was observed.

Animals↗

CD4+ T cells determine the ability of spleen cells from F1 hybrid mice to induce neonatal tolerance to alloantigens and autoimmunity in parental mice.

Spleen cells from F1 hybrid mice injected into newborn parental mice induce a state of cytolytic unresponsiveness to the corresponding alloantigens. However, these mice develop a transient autoimmune syndrome characterized by the production of multiple autoantibodies and glomerulonephritis. Previous reports indicated that the depletion of F1 donor T cells, shortly prior the injection into parental mice, does not interfere with any of these events. Here, we have explored whether the continuous absence of T cells in F1 mice influences the ability of their spleen cells to induce neonatal tolerance to alloantigens and the associated autoimmune manifestations. Our results revealed that spleen cells from athymic (BALB/c x C57BL/6) F1 hybrid (CB6F1) nu/nu mice or from euthymic CB6F1 mice depleted from birth of CD4+ T cells, but not of CD8+ T cells, are unable to induce neonatal tolerance to alloantigens and autoimmune manifestations. By contrast, the partial reconstitution of T cells in CB6F1 nu/nu mice, after the neonatal graft of a syngeneic thymus, restored the capacity of spleen cells from these mice to induce tolerance and autoimmunity when injected into newborn BALB/c mice. These results demonstrate that the functional defect of spleen cells from athymic CB6F1 nu/nu mice to induce neonatal tolerance to alloantigens is directly related to the long-term absence of mature CD4+ T cells. Interestingly, a new increase in the titers of anti-DNA Ab was observed when spleen cells from athymic CB6F1 nu/nu mice were injected into adult BALB/c mice that had been tolerized at birth with normal CB6F1 spleen cells. This finding indicates that B cells from CB6F1 nu/nu mice recover their capacity to interact with alloreactive Th2 cells when they are placed into mice having functional CD4+ T cells. These data indicate that the continuous absence of CD4+ T cells causes a reversible functional defect in F1 spleen cells that determines their inability to induce neonatal tolerance and autoimmunity.

Animals↗

Induction of sister chromatid exchange in spleen and bone marrow cells of rats exposed by inhalation to different dose rates of ethylene oxide.

We investigated the effects of dose rate on the frequency of sister chromatid exchange (SCE) in bone marrow and spleen cells of rats exposed to ethylene oxide (EtO). Four groups (18/group) of male Fischer 344 rats were exposed to EtO by inhalation. The exposures consisted of 100 ppm for 6 hr/day, 300 ppm for 2 hr/day, 600 ppm for 1 hr/day, and clean air control. All EtO treated rats were given a total exposure dose of 600 ppm.hr daily, 5 days/week for 3, 6, or 9 months. Six rats per group were sacrificed at each time point, and SCEs were measured in cultured spleen and bone marrow cells. A statistically significant increase was found in SCEs in both bone marrow and spleen cells for all treated groups and at each time point when compared to the control, except at the 3-month exposure for the middle and high dose-rate groups in bone marrow cells. In the spleen, the increases in SCEs were similar among the three experimental groups. In bone marrow, the lowest dose rate (100 ppm) resulted in higher SCE frequencies than the medium and high dose-rate group after 3 and 6 month exposures. The overall frequencies of SCEs in the spleen cells were higher than in the bone marrow cells. The increase in SCE frequencies and decrease in the replicative index in spleen cells were also dependent on the duration of exposure. These results indicate that (1) EtO, by inhalation, can cause SCEs both in spleen and bone marrow cells of Fischer 344 rats, (2) spleen cells are more sensitive to EtO than bone marrow cells, and (3) in bone marrow cells the lowest dose-rate (longest) exposure causes more SCEs than the highest dose-rate (shortest) exposures.

Administration, Inhalation↗

Cell-mediated cytotoxicity of sensitized spleen cells against target liver cells--in vivo and in vitro study with a mouse model of experimental autoimmune hepatitis.

Spleen cells obtained from C57BL/6 (B6) mice with an experimental autoimmune hepatitis were transferred to normal C57BL/6 recipient mice. Most prominent liver damages occurred in the recipient mice injected with sensitized nylon wool column-adherent spleen cells from the donor mice. Production of such liver damage was blocked by treatment of the sensitized adherent spleen cells with anti-Thy 1,2 monoclonal antibody and complement before injection. Based on these in vivo results, a microcytotoxicity assay was performed using isolated C57BL/6 hepatocytes as target cells and sensitized spleen cells obtained from hepatitis donor mice as effector cells. The fraction of sensitized nylon wool-adherent spleen cells demonstrated a high cytotoxic activity against isolated syngeneic hepatocytes, although the other fractions and spleen cells of control animals showed no such effect. The cytotoxic activity of sensitized-adherent spleen cells against target hepatocytes was significantly reduced after treatment with anti-Thy 1,2 antibody and complement, but it increased after depletion of B cells and Fc receptor-bearing T-cells. Although these sensitized nylon wool-adherent spleen cells showed high cytotoxic activities against syngeneic hepatocytes, their cytotoxicity against allogeneic hepatocytes was lower. They exerted no cytotoxic activity against syngeneic renal glomerular cells and EL-4 thymoma cells. These results suggest that sensitized T-cells in the nylon wool column-adherent fraction play the role of cytotoxic killer cells against target liver cells in vitro.

Animals↗

Sonographic assessment and grading of spleen size.

Ultrasonography was used to evaluate spleen size in patients with various clinical conditions including those of the liver, blood, collagen, or autoimmune disease. To express spleen size, a spleen index (SI), the product of the transverse diameter and its perpendicular diameter measured on the maximum cross-sectional image of the spleen, was used. SI correlated well with the volumes of resected spleens. Splenomegaly was present in high percentages of patients with liver, blood, collagen, and autoimmune disease, even though a majority of these spleens were not large enough to palpate. By grading the SI, characteristic distributions of SI were obtained for patients with different types of diseases. Obtaining and grading the SI by the use of ultrasound appears to be a significant supplemental aid for evaluating spleen size, especially in patients whose spleens are not palpable.

Adolescent↗

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.

Animals↗

Interferon gamma priming is not critical for IL-12 production of murine spleen cells.

Interferon gamma (IFN-gamma) priming is considered to be critical for interleukin 12 (IL-12) production of murine macrophages and human monocytes by lipopolysaccharide (LPS) stimulation. In our present experiments, freshly prepared spleen cells (f-spleen cells) were confirmed not to produce detectable level of IL-12 by LPS stimulation, although they produced significant amount of IL-12 by the stimulation with LPS plus IFN-gamma. However, the stimulation only with LPS induced IL-12 production of spleen cells preincubated in the absence of IFN-gamma. Findings on IL-12 p40 mRNA accumulation were consistent with their IL-12 production. Essentially the same results were obtained using spleen cells from IFN-gamma deficient mice. In the presence of anti-IL-10, f-spleen cells produced IL-12 upon LPS stimulation, indicating that the failure of f-spleen cells in IL-12 production is caused by IL-10 produced by themselves upon LPS stimulation. In addition, f-spleen cells produced IL-12 upon CD40 ligand stimulation, and the production was hardly affected by the presence of IFN-gamma or preincubation. These results indicate that IFN-gamma priming is not critical for IL-12 production of spleen cells stimulated with LPS or CD40 ligand, although IFN-gamma enhances the production, especially, in response to LPS stimulation.

Animals↗

Suppression of antitumor immunity by macrophages in spleens of mice bearing a large MOPC-315 tumor.

We had shown previously that progression of MOPC-315 plasmacytoma growth is associated with an increase in the percentage of macrophages in the spleen as well as a decrease in the ability of tumor-bearer spleen cells to mount an antitumor cytotoxic response upon in vitro immunization. Here we provide evidence that macrophages in the MOPC-315 tumor-bearer spleen are responsible at least in part for the suppression of the generation of antitumor cytotoxicity. Accordingly, removal of most macrophages by depletion of phagocytic cells or Sephadex G-10-adherent cells from spleens of mice bearing a large tumor resulted in augmented antitumor immune potential. Also, Sephadex G-10-adherent spleen cells from tumor-bearing (but not normal) mice drastically suppressed the in vitro generation of antitumor cytotoxicity by normal spleen cells. The suppressive activity of these adherent cells did not reside in contaminating suppressor T cells, since it was not reduced by treatment with monoclonal anti-Thy 1.2 antibody plus complement. The Sephadex G-10-adherent cell population from the tumor-bearer spleen suppressed the in vitro generation of antitumor cytotoxicity against autochthonous tumor cells but not against allogeneic EL4 tumor cells, and hence the suppression was apparently specific. The suppressive activity of the Sephadex G-10-adherent cell population from tumor-bearer spleens was overcome by treatment of the tumor-bearing mice with a low curative dose of cyclophosphamide. This immunomodulatory effect of a low dose of the drug in overcoming the suppression mediated by the Sephadex G-10-adherent cell population enables the effector arm of the immune system of tumor-bearing mice to cooperate effectively with the drug's tumoricidal activity in tumor eradication.

Animals↗

Laparoscopic splenopexy by peritoneal and omental pouch construction for intermittent splenic torsion ("wandering spleen").

Wandering spleen is an extremely rare anatomic variant with potentially serious clinical implications. Usually, splenectomy is advocated for treatment of this disease. Various methods for preserving the wandering spleen by means of splenopexy have been described, including two reports on laparoscopic splenic refixation. We describe the third case in which laparoscopic splenopexy was used to manage chronic intermittent splenic torsion. In a 25-year-old woman, splenopexy was successfully performed by laparoscopic reposition and fixation of the spleen by omental pouch creation. At laparoscopy with a normal operating room setup and four trocars, a free-floating, macroscopically normal spleen attached to an abnormally long vascular pedicle with no gastrosplenic or phrenicosplenic ligaments was detected in the lower right quadrant. The spleen was repositioned and placed in the left phrenorenal angle. Splenopexy was achieved by suturing the left colophrenic ligament to the lateral diaphragm, thus creating a pouch for the inferior part of the spleen, and by suturing the gastrocolic ligament to the anterior diaphragm to create a pouch for the upper splenic pole. The postoperative course was uneventful. At a follow-up examination 3 months after the operation, the patient was well, with no further episode of recurrent abdominal pain. Ultrasonographically, the spleen was seen easily in the left hypochondrium in its normal physiologic position. Laparoscopic splenopexy is a useful option for organ-preserving therapy of the wandering spleen.

Adult↗

Inadequate detection of accessory spleens and splenosis with laparoscopic splenectomy. A shortcoming of the laparoscopic approach in hematologic diseases.

BACKGROUND: The ultimate goal of surgery for hematological disorders is the complete removal of both the spleen and accessory spleens in order to avoid recurrence of the disease. Whereas splenectomy by open surgery provides excellent results, the validity of laparoscopic splenectomy in this regard remains unknown. OBJECTIVE: The purpose of this study was to evaluate the detection of accessory spleens during laparoscopic splenectomy for hematologic diseases. METHODS: We therefore evaluated the pre-, intra-, and postoperative detection of accessory spleens in a consecutive series of 18 patients treated by elective laparoscopic splenectomy for hematological diseases by using computed tomography (CT) and denatured red blood cell scintigraphy (DRBCS). RESULTS: Preoperative CT, DRBCS, and laparoscopic exploration detected 25%, 25%, and 75% of accessory spleens, respectively. At time of laparoscopy, 16 accessory spleens were detected in seven of the 18 patients (41%). In two patients (11%), laparoscopic exploration failed to detect accessory spleens, whereas preoperative CT (one case) and DRBCS (one case) did reveal them. Postoperatively, during a mean follow-up of 28 months (median, 24; range, 12-44 months), nine patients (50%) showed persistence of splenic tissue by DRBCS, and three of them had signs of disease recurrence. CONCLUSIONS: This prospective clinical study suggests that elective laparoscopic surgery for hematological diseases does not allow complete detection of accessory spleens. Moreover, after such a laparoscopic approach, residual splenic tissue is detectable in half of the patients during the follow-up.

Adolescent↗

Peritoneal "pillowcase" for the displaced spleen post-distal pancreatectomy.

BACKGROUND PURPOSE: Spleen-preserving distal pancreatectomy, although rarely performed, has become an option for treatment of distal pancreatic neoplasms. However, the spleen, once mobilised, is often free to tort. A reliable method of fixing the spleen would be a useful adjunct to this procedure. METHODS: We describe the use of a new technique to fix the spleen using a peritoneal "pillowcase" to envelop the spleen, thus preventing its torsion. RESULTS: This patient has done well, with no problems encountered intraoperatively or postoperatively, using this new technique of spleen fixation. CONCLUSIONS: The use of a peritoneal "pillowcase" to envelope the spleen following distal pancreatectomy has proven a useful adjunct to this somewhat difficult surgical procedure. It may be applied in other cases where spleen fixation is necessary.

Cystadenocarcinoma↗