The aim of study was to investigate the effectiveness of allogeneic natural killer (NK) cells in haploidentical bone marrow transplantation (BMT) for leukemia mice. CB6F(1)(H-2b/d) murine model of EL9611 (H-2d) erythroleukemia was established by intravenous injection of EL9611 (H-2(d)) cells. CB6F(1)(H-2b/d) mice were transplanted with bone marrow (BM) cells from C57BL/6(H-2b) mice. Seventy CB6F(1)(H-2b/d) mice were randomly divided into 7 groups with 10 mice per group. 5 control groups were: group 1, in which no treatment was performed; group 2, in which mice were lethally irradiated (9 Gy); group 3, in which mice were treated with cytarabine with dose of 50 mg/kg for 6 days followed by the infusion of EL9611 (H-2(d)); group 4, in which mice were transplanted with BMT and group 5-the GVHD-control group, in which mice were transplanted with BM and spleen cells from C57BL/6(H-2b) mice 4 hours after irradiation. Experimental groups were divided into 2 groups: group A, in which mice were injected with C57BL/6(H-2b) NK cells (1 x 10(6)) after irradiation and were transplanted with BM from C57BL/6(H-2b) 4 hours later, and group B, in which mice were transplanted with BM cells and spleen cells from C57BL/6(H-2b) 4 hours after irradiation. The effect was assessed and compared by blood picture, survival time, body weight, and histopathology in the recipients. The results showed that the survival times in control group 1, 2, 3 and 5 were (10.10 +/- 0.88), (9.80 +/- 0.92), (22.70 +/- 3.23) and (20.10 +/- 1.73) days respectively. The survival time of control group 4 was (30.10 +/- 15.95) days and was over 30 days in 2 mice. The survival times in experimental group A and B were (39.10 +/- 18.11) and (49.30 +/- 17.24) days respectively. 4 mice in experimental group 1 and 7 mice in group 2 survived over 30 days. The survival time of experimental group 1 was significantly longer than that of control group 1, 2, 3 and 5 (p < 0.01). The survival time of experimental group 2 was significantly longer than that of other groups (p < 0.05). Histopathologic examination showed that splenohepatomegalia and disorganization of liver and spleen with infiltration of a large amount of leukemia cells in mice dead of leukemia. Chimerism of Y chromosome was shown in mice of experimental groups with long survival time. It is concluded that the injection with donor-derived NK cells can both eliminate leukemia cells and decrease the severity of GVHD after haploidential BMT.
Animals ; Bone Marrow Transplantation ; methods ; Killer Cells, Natural ; transplantation ; Leukemia ; surgery ; Mice ; Mice, Inbred C57BL ; Spleen ; transplantation ; Transplantation, Homologous

OBJECTIVETo evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat augmenter of liver regeneration (rALR) injection in treating rats with acute hepatic failure.

METHODSAcute hepatic failure (AHF) was induced in rats using D-gal (1.2 g/kg). The rats were then randomly divided into 6 groups. Group I received no further treatment and served as blank controls; group II received 1 ml buffered saline once through intrasplenic injection; group III received 1 ml rALR; group IV received 2 x 10(7)/ml hepatocytes; group V received 2 x 10(7) hepatocytes suspended in 1 ml rALR (50 microg/kg) and group VI received 2 x 10(7) hepatocytes in 1 ml cyclosporine A (10 mg/kg). The rats of the various treated groups were sacrificed at day 1, 5 and at week 2 and their livers and spleens were examined histopathologically. Blood samples of the rats were also obtained to determine the levels of TNF alpha and IL-1 beta.

RESULTSThere were no significant differences in survival between group I, II and III rats. 33.3% of the group IV rats survived for 2 weeks. At week 2, the survival rate of group V rats was significantly higher than that of group IV, but there was of no statistical significant increase when compared to that of group VI rats. Hepatocytes transplanted into spleens survived for 5-7 days in the spleens of group IV and VI rats, but they survived at least 2 weeks in group V. The average serum TNF alpha level in group IV was significantly higher than that in groups V and VI on the first postoperative day, but after four days, only the difference between group IV and group VI was significant (P < 0.05). The average serum level of TNF alpha in group II was higher than that in groups IV, V and VI on the first postoperative day (P < 0.05), but there were no significant differences between those in groups IV, V and VI on the 1st and the 5th postoperative days.

CONCLUSIONNewborn rat hepatocyte intrasplenic transplantation combined with rALR is effective in treating acute hepatic function failure induced by D-gal in rats. The transplanted hepatocytes can survive for at least 2 weeks in the spleens. The rALR mixed with the hepatocytes injected into the spleens may be able to facilitate the hepatocyte regeneration, to inhibit liver cell apoptosis and to suppress the cellular immunity.

Animals ; Cell Transplantation ; methods ; Female ; Hepatocytes ; transplantation ; Liver Failure, Acute ; immunology ; surgery ; Liver Regeneration ; Male ; Proteins ; therapeutic use ; Rats ; Rats, Wistar ; Spleen ; surgery

This study was conducted by consecutively transplanting spleens, which had gonads implanted previously. A total of 84 cases for infantile testicles and 106 cases for ovarian follicles were performed. In the case of ovarian implants, the results were determined by the total number of follicle implants. A modified spleen transplantation technique called double implantation of ovarian follicles was applied to increase the amount of the implants. In this technique, an extra spleen is implanted into the potential donor so that the ovarian follicles can be implanted to two different spleens, doubling the amount of implants. Through consecutive spleen transplantation, we observed the results beyond a typical rat's life span. In many of these cases, we found more aggressive forms of malignant tumor, seminoma and dysgerminoma. We present the results and discuss possible pathogenic mechanisms of tumor formation.
Animals ; Animals, Newborn ; Female ; Male ; Ovarian Neoplasms/*etiology ; Ovary/*transplantation ; Rats ; Rats, Inbred Lew ; Research Support, Non-U.S. Gov't ; Spleen/*surgery/*transplantation ; Testicular Neoplasms/*etiology/pathology ; Testis/*transplantation ; *Transplantation, Heterotopic

OBJECTIVETo study the therapeutic effect of splenic autotransplantation combined with lower esophagus transaction anastomosis in the treatment of liver cirrhosis induced portal hypertension.

METHODSThirty-six patients admitted from January 2003 to December 2006 were randomly divided into splenic autotransplantation group undergoing splenic autotransplantation after splenectomy combined with lower esophagus transaction anastomosis, and splenectomy group only undergoing splenectomy combined with lower esophagus transaction anastomosis. The general conduction, splenic scanning, liver function, and the level of serum Tuftsin and IgM of each patient were observed before and after operation.

RESULTSThe levels of Tuftsin and IgM in splenic autotransplantation group were significant higher than that of splenectomy group 2 months after the operation, and the liver function showed no significant difference between these two groups. Splenic tissue was detected in the retroperitoneal space by 99mTc-DRBC 2 months after operation.

CONCLUSIONSSplenic autotransplantation combined with lower esophagus transaction anastomosis is a safe and effective treatment strategy for patients with liver cirrhosis induced portal hypertension, and the spleen tissue transplanted into the retroperitoneal space can partially preserve the immune function.

Adult ; Aged ; Aged, 80 and over ; Anastomosis, Surgical ; Esophagus ; surgery ; Female ; Humans ; Hypertension, Portal ; etiology ; surgery ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Spleen ; transplantation ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

BACKGROUNDSurgical treatment options for patients with cirrhosis and portal hypertension are complicated. In this study, we evaluated the effectiveness of a new treatment strategy, splenic auto-transplantation and oesophageal transection anastomosis. We report results from clinical observations, splenic immune function and portal dynamics in 274 patients.

METHODSFrom 1979 to 2005, 274 cirrhosis patients with portal hypertension underwent the new treatment strategy, and were followed up to compare results with those patients who underwent traditional surgical treatment. From 1999 to 2002, a randomized controlled trial (RCT) was performed on 40 patients to compare their post-operative immune function. From 1994 to 2006, another RCT enrolled 28 patients to compare portal dynamics using three-dimensional dynamic contrast-enhanced magnetic resonance angiography (3D DEC MRA) investigation post operation.

RESULTSAmong 274 patients (mean age 41.8 years), the emergency operative mortality (4.4%), selective operative mortality (2.2%), complication rate (17.9%), prevalence of hepatic encephalopathy (< 1%), rate of portal hypertension gastritis (PHG) bleeding (9.1%), and morbidity of hepatic carcinoma (8%) were similar to those patients undergoing traditional operation; the spleen immunology function (Tuftsin, IgM) decreased in both groups 2 months post operation, but this decrease did not reach statistical significance. Through 3D DCE MRA, the cross sectional area and the velocity and volume of blood flow of the main portal vein decreased significantly after operation in both groups. The velocity and volume of blood flow in the auto-transplantation group was significantly lower than that in the control group.

CONCLUSIONSSplenic auto-transplantation and esophageal transection anastomosis is a safe, effective, and reasonable treatment strategy for patients with portal hypertension with varicial bleeding. It not only can correct hypersplenism, but may also achieve complete hemostasis. Spleens auto-transplanted into the retroperitoneal space can preserve immune function and establish broad collateral circulation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anastomosis, Surgical ; Child ; Esophagus ; surgery ; Female ; Humans ; Hypertension, Portal ; immunology ; surgery ; Imaging, Three-Dimensional ; Immunoglobulin M ; blood ; Male ; Middle Aged ; Prospective Studies ; Spleen ; transplantation ; Transplantation, Autologous

OBJECTIVETo observe the in vivo colonization, migration, and differentiation of in vitro cultured human fetal hepatic stem cells (HSCs) following intrasplenic transplantation for treatment of acute liver injury in mice with severe combined immunodeficiency (SCID).

METHODSHuman fetal HSCs were isolated from the normal fetal liver (16-24 weeks) and purified, and the morphology of HSCs was observed under optical and transmission electron microscopes. The expressions of stem cell markers were examined in these HSCs by means of immunocytochemistry and flow cytometry. The passaged human fetal HSC suspension (0.2 ml) were injected into the spleen of SCID mice with acute liver injury induced by two-third partial hepatectomy, and 15, 30, 60, and 90 days after cell transplantation, immunohistochemistry was performed to examine the location and expressions of human hepatocytes, alpha1-AT and AFP antigen in the spleen and liver of the recipient SCID mice. PAS staining was used to examine the expression of glycogen and RT-PCR employed for detection of the expressions of AFP and albumin mRNA in the spleen of the mice on the scheduled time points.

RESULTSUnder optical microscope and transmission electron microscope, most of the HSCs were small, about 1/6 to 1/3 of the size of the hepatocyte, with relatively large nucleus-cytoplasm ratio and only small quantities of endocytoplasmic reticulum, chondriosome, and ribosome. Immunohistochemistry and flow cytometry identified positive expressions of AFP, Thy-1, C-kit, CD34 and CK19 in the HSCs, and after cell transplantation, positive expressions of human hepatocyte, alpha1-AT, and AFP antigen occurred in the liver and spleen of the recipient SCID mice. PAS staining confirmed the presence of glycogenosome in the spleen of the mice following cell transplantation. RT-PCR on days 30, 60, and 90 showed positive expressions of human AFP and albumin mRNA in the spleen of the mice.

CONCLUSIONHuman fetal HSCs can survive and settle in the spleen and liver, and migrate to the damaged liver of the recipient mice after intrasplenic transplantation, with the capacity of proliferation and differentiation into hepatocytes in the recipient target organs.

Animals ; Cells, Cultured ; Female ; Fetal Stem Cells ; cytology ; transplantation ; ultrastructure ; Flow Cytometry ; Hepatectomy ; methods ; Hepatocytes ; cytology ; metabolism ; transplantation ; Humans ; Immunohistochemistry ; Liver ; injuries ; surgery ; Mice ; Mice, Inbred BALB C ; Mice, SCID ; Microscopy, Electron, Transmission ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; metabolism ; surgery ; Stem Cell Transplantation ; methods ; Transplantation, Heterologous ; alpha-Fetoproteins ; analysis ; genetics

OBJECTIVETo study the regularity of splanchnic hemodynamic changes after orthotopic liver transplantation (OLT) for patients with portal hypertension. At the same time, effect of such changes on splenomegaly, hypersplenism, collateral circulation and the postoperative liver function was discussed.

METHODSBetween June 2002 and October 2005, 173 liver transplantations were performed. In 38 patients with portal hypertension undergoing OLT, the following parameters were measured before surgery and subsequently at 1, 3, 5, 7 days, 1, 6 months and 1, 2, 3 years after operation by using Color Doppler sonography: portal blood flow mean velocity (PBV), portal blood flow volume (PBF), hepatic artery resistance indexes (HA-RI) and spleen size. The same parameters were measured in 8 patients with acute liver failure and 20 healthy controls. Meanwhile to observe liver function and varicose vein of esophagus.

RESULTSIn cirrhotics, PBV and PBF increased immediately after transplantation [from (13.7 +/- 4.2) cm/s to (58.4 +/- 25.2) cm/s and from (958 +/- 445) ml/min to (3024 +/- 1207) ml/min respectively, P < 0.05]. HA-RI also augmented [from (0.65 +/- 0.11) to (0.74 +/- 0.12), P < 0.05]. PBV returned to normal values after 6 months, PBF returned to normal value after 2 years. Spleen size decreased significantly, but splenomegaly persisted after 3 years. In addition the esophagogastric varix ameliorated significantly.

CONCLUSIONSAbnormal splanchnic hemodynamic changes for patients with portal hypertension still will long-term exist after OLT, but does not effect recovery of hypersplenism, esophagogastric varix and liver function.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Hemodynamics ; Hepatic Artery ; physiopathology ; Humans ; Hypertension, Portal ; pathology ; physiopathology ; surgery ; Intraoperative Period ; Liver ; physiopathology ; Liver Transplantation ; Male ; Middle Aged ; Portal Vein ; physiopathology ; Splanchnic Circulation ; physiology ; Spleen ; pathology

OBJECTIVETo discuss the risk factors of splenic arterial steal syndrome (SASS) after orthotopic liver transplantation.

METHODSTwenty-four cases who confirmed SASS after liver transplantation in Tianjin First Central Hospital between June 2005 and June 2013 were analyzed retrospectively. Another 96 cases were selected randomly from those patients of the same time with no complication of SASS patients postoperatively as control group. Clinical data of two groups including diameter of splenic artery and hepatic artery preoperatively, weight of graft, weight of recipients, cold/warm ischemia time, an hepatic period and operation time and so on were collected. Others including hepatic artery peak systolic velocity (PSV), end diastolic velocity (EDV), blood flow resistance index and portal vein average velocity (PVF) on the first day after liver transplantation, the day before diagnosis, the day when diagnosed, the 1, 3, 7 days after treatment in SASS group and on 1, 3, 7, 9, 11, 14 days after liver transplantation in control group. Statistical analysis were made between two groups.

RESULTSThe splenic artery/hepatic artery ratio preoperatively and weight of donor liver,and the GRWR in SASS group and control group were 1.26 and 1.00, 1 032 g and 1 075 g, (1.40±0.30)% and (1.82±0.21)% respectively, with significantly statistical differences (Z=-6.40, Z=-2.22, t=-6.50; all P<0.05). The warm ischemia time, the cold ischemia time, the anhepatic period and operation time in SASS group and control group were 3.5 minutes and 4.0 minutes, 10.25 hours and 10.10 hours, 43 minutes and 45 minutes, 8.7 hours and 8.7 hours, with no significantly statistical differences (all P>0.05). RI of hepatic went up gradually in the early time after transplantation while dropped obviously when spleen artery spring coils embolization was received (P<0.01) and trended to stable two weeks later.

CONCLUSIONSSplenic artery/hepatic artery ratio and GRWR are the positive and negative risk factors respectively for SASS. The gradual rising of hepatic RI in the early time after transplantation may be the warning signal SASS and spleen artery spring coils embolization is the effective strategy for SASS after liver transplantation.

Cold Ischemia ; Embolization, Therapeutic ; Hepatic Artery ; pathology ; Humans ; Liver ; surgery ; Liver Transplantation ; adverse effects ; Retrospective Studies ; Risk Factors ; Spleen ; blood supply ; Splenic Artery ; pathology ; Vascular Diseases ; epidemiology ; Warm Ischemia

OBJECTIVETo observe the evolution and differentiation of hepatic oval cells after transplanted into the spleens of homogenous rats, providing experimental data for treating hepatic failure with hepatic stem cells.

METHODSA two-step perfusion procedure was used to separate hepatic parenchymal cells from nonparenchymal cells. Then the suspension of nonparenchymal cells was centrifuged in Percoll gradients. The isolated cells were cultured, identified, and then transplanted into the spleens of homogenous rats undergone 2/3 hepatectomy.

RESULTSThe obtained cells were various in size with ovoid nuclei and inadequate cytoplasm. After 12 hours' culture, they revealed the characteristics of epithelial cells. Both the freshly isolated and cultured cells showed positive staining for cytokeratin 19 (CK19), OV6, alpha fetal protein (AFP), but negative for leucocyte common antigen (LCA). After intraspleenic transplantation into homogenous rats undergone partial hepatectomy, hepatic oval cells were differentiated into liver tissue-like structure including hepatocyte cords and bile ducts, and formed hepaticized spleen. But this kind of structure was not observed in the controls.

CONCLUSIONThe isolated rat hepatic oval cells show the biological characteristics of hepatic stem cells and can differentiate into hepatocytes and biliary epithelial cells under appropriate circumstances.

Animals ; Antigens, Differentiation ; analysis ; Cell Differentiation ; Cell Separation ; Cell Transplantation ; pathology ; Cells, Cultured ; Hepatectomy ; methods ; Hepatocytes ; drug effects ; physiology ; ultrastructure ; Liver ; cytology ; growth & development ; Liver Transplantation ; methods ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Spleen ; surgery ; Stem Cells ; drug effects ; physiology ; ultrastructure

OBJECTIVETo study the effect of pretreatment with apoptotic donor spleen cells on spleen lymphocyte function of recipient rats undergoing islet transplantation to explore new approaches to prolong islet graft survival.

METHODSApoptotic spleen cells from donor rats were obtained by exposure to γ-ray irradiation from (60)Co. Diabetic SD rat models were randomly divided into 4 groups to receive tail vein injections with saline (group A), normal cells (group B), apoptotic donor cells (group C), or necrotic donor cells (group D). One week later, orthotopic transplantation of islets under the renal capsule was performed. Before and at 1 and 2 weeks after islet transplantation, the recipient rats were examined for proliferative activity of spleen lymphocytes with CFSE cell staining and for IL-2 and IL-10 expressions in the cells using ELISA.

RESULTSPretreatment with donor apoptotic cells significantly suppressed the proliferative activity of recipient spleen lymphocytes before and at 1 and 2 weeks after islet transplantation as compared with the other three groups (P<0.05). The level of IL-2 was significantly decreased while IL-10 increased in apoptotic donor cell pretreatment group compared with those in the other 3 groups at each time point of observation.

CONCLUSIONThe effect of pretreatment with apoptotic donor cells on recipient spleen lymphocytes suggest an important role of apoptotic donor spleen cells in immune tolerance of grafts.

Animals ; Apoptosis ; immunology ; Cell Proliferation ; Cobalt Radioisotopes ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; surgery ; Gamma Rays ; Graft Survival ; Immune Tolerance ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Islets of Langerhans Transplantation ; Lymphocyte Transfusion ; Lymphocytes ; metabolism ; pathology ; radiation effects ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Spleen ; cytology ; metabolism ; radiation effects