Camptothecin/*pharmacology ; Liver Neoplasms, Experimental/*immunology ; Neoplasm Transplantation ; Sarcoma, Experimental/*immunology ; Transplantation, Isogeneic

Animals ; Camptothecin ; pharmacology ; Liver Neoplasms, Experimental ; immunology ; Mice ; Neoplasm Transplantation ; Sarcoma, Experimental ; immunology ; Transplantation, Isogeneic

BACKGROUND: Rotor off (R/O) fraction obtained by counterflow centrifugal elutriation (CCE) contains small number of T cells and many hematopoietic stem cells. Since megakaryocytes and its progenitors are larger than other cells in bone marrow, it may be easier to be separated by CCE and newly applied to hematopoietic stem cell transplantation (HSCT) for early megakaryocytes reconstitution. METHODS: The marrow cells of BALB/c mice in each group (17, 25, 28mL/min, and R/O fraction) were cultured for quantifying CFU-MK and measured after 10 days. BALB/c mice were lethally irradiated and transplanted with R/O cells. The dosages of transplanted cells were 5x104 in Group A, 5x105 in Group B, and 5x106 in Group C. The platelet counts in peripheral blood were measured up to post-transplant day 14. RESULTS: After CCE, recovery rate of the loaded cells was 82.2% and the R/O fraction was 35.9%. Most CFU-MK were formed in R/O fraction, and Group C showed the fastest recovery. Group A couldn't reach the level of 100x109/L until post-transplant day 14, and Group B showed slower recovery compared to Group C. All 5 mice survived in Group C, but 2 out of 5 mice survived in Group A and B. CONCLUSIONS: R/O fraction contains higher number of megakaryocyte progenitors, and CCE could be an effective method for separating megakaryocyte progenitors essential for reconstituting platelet after HSCT. The cell dose of 5x106 was required for the effective recovery of platelet and the survival of BALB/c mice in syngeneic bone marrow transplantation with R/O cells.
Animals ; Blood Platelets* ; Bone Marrow ; Bone Marrow Transplantation ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Megakaryocyte Progenitor Cells ; Megakaryocytes ; Mice* ; Platelet Count ; T-Lymphocytes ; Transplantation, Isogeneic*

OBJECTIVETo establish stoma and stoma-free murine models of heterotopic small intestine transplantation in order to choose a more effective and reliable model.

METHODSA total of 140 male 8-10 weeks age C57BL/6(B6) mice weighted 25-30 g were enrolled in the experiment. Syngeneic heterotopic small intestine transplantation was performed between C57BL/6 mice, and recipient mice were divided into either stoma or stoma-free group. Heterotopic small intestine transplantation was performed in 70 mice, with 35 mice in each group. After closing the proximal end of the graft by ligation, the distal end of graft was exteriorized as a stoma then secured to the skin of the abdominal wall in stoma group. In stoma-free group, the distal end of graft was anastomosed end-to-side to the recipient ileum. Successful rate of operation, two-week survival rate, operation time, associated complications, postoperative care time and body weight change were recorded and compared between two groups.

RESULTSThe successful rate of stoma group was 65.7%, while it was 80.0% of stoma-free group (χ(2)=1.806, P=0.179). The operation time of donor in stoma group was (48.1±6.6) minutes, while it was (47.2±5.9) minutes in stoma-free group (t=0.598, P=0.552). The operation time of recipient in stoma group was (77.9±9.1) minutes, while it was (76.4±8.3) minutes in stoma-free group (t=0.683, P=0.497). The cold ischemic time of graft in stoma group was (34.7±4.0) minutes, while it was (33.9±4.6) minutes in stoma-free group(t=0.667, P=0.507). The two-week survival rate of stoma group was 45.7%, and it was 77.1% of stoma-free group(χ(2)=7.295, P=0.007). The stoma group had more complications[54.3%(19/35) vs. 22.9%(8/35), χ(2)=7.295, P=0.007], which needed more postoperative care time(191 min vs. 35 min). The weight loss in stoma group in the third day after operation was more significant [(81.52±5.20)% vs. (85.46±4.65)%, t=2.856, P=0.006]. By 2 weeks after operation, the weight of mice in both groups retruned to 95% of the postoperative wight.

CONCLUSIONThe murine heteropotic small intestine transplantation model with stoma-free appears to be more reasonable and reliable.

Animals ; Digestive System Surgical Procedures ; Ileum ; surgery ; Intestine, Small ; transplantation ; Male ; Mice ; Mice, Inbred C57BL ; Surgical Stomas ; Transplantation, Heterotopic ; methods ; Transplantation, Isogeneic

An orthotopic penetrating keratoplasty model was developed in the rat. An oversized (0.5 mm) graft was used and 8 interrupted sutures were applied. These sutures were not removed. Eleven grafts out of 13 were rejected by the 3rd week in the disparate group (Brown Norway rat to Lewis rat transplantation group), which was characterized by edema, opacity, and neovascularization. All grafts remained clear in the syngeneic group (Lewis rat to Lewis rat transplantation group). Immunohistochemical examination was performed. This model seems to be a reliable and reproducible one to evaluate rejection reaction in corneal transplantation.
Animals ; Female ; Graft Rejection ; Keratoplasty, Penetrating/immunology/*methods/pathology ; Lymphocyte Subsets/immunology/pathology ; Macrophages/immunology/pathology ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Transplantation, Homologous ; Transplantation, Isogeneic

The objective of this study was to investigate the effect of ligustrazine on the expression of stem cell factor mRNA (SCF) in bone marrow tissue and explore the mechanism of hematopoietic reconstitution after bone marrow transplantation (BMT). The colony forming unit of spleen (CFU-S) were counted, the survival rate at days 7, 14 and 21 after BMT were measured, as well as the expression level of SCF mRNA was detected by RT-PCR. The results showed that in ligustrazine group CFU-S counts on day 10 and survival rate, expression level of SCF mRNA on day 7, 14 and 21 after BMT were higher than that in the control group (p < 0.01 or p < 0.05). In conclusion, ligustrazine promotes the recovery of hematopoietic cells in bone marrow, enhances the repair of bone marrow microvessels, and then improves bone marrow microenvironment and promotes hematopoietic reconstitution.
Animals ; Bone Marrow Transplantation ; Hematopoiesis ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Stem Cell Factor ; genetics ; metabolism ; Transplantation, Isogeneic

BACKGROUNDA practical problem impeding clinical translation is the limited bone formation seen in artificial bone grafts. Low-pressure/vacuum seeding and dynamic culturing in bioreactors have led to a greater penetration into the scaffolds, enhanced production of bone marrow cells, and improved tissue-engineered bone formation. The goal of this study was to promote more extensive bone formation in the composites of porous ceramics and bone marrow stromal cells (BMSCs).

METHODSBMSCs/β-tricalcium phosphate (β-TCP) composites were subcultured for 2 weeks and then subcutaneously implanted into syngeneic rats that were split into a low-intensity pulsed ultrasound (LIPUS) treatment group and a control group. These implants were harvested at 5, 10, 25, and 50 days after implantation. The samples were then biomechanically tested and analyzed for alkaline phosphate (ALP) activity and osteocalcin (OCN) content and were also observed by light microscopy.

RESULTSThe levels of ALP activity and OCN content in the composites were significantly higher in the LIPUS group than in the control group. Histomorphometric analysis revealed a greater degree of soft tissue repair, increased blood flow, better angiogenesis, and more extensive bone formation in the LIPUS groups than in the controls. No significant difference in the compressive strength was found between the two groups.

CONCLUSIONLIPUS treatment appears to enhance bone formation and angiogenesis in the BMSCs/β-TCP composites.

Animals ; Bone Marrow Cells ; physiology ; Bone Transplantation ; Calcium Phosphates ; pharmacology ; Male ; Osteogenesis ; physiology ; Rats ; Stromal Cells ; Tissue Engineering ; methods ; Transplantation, Isogeneic ; Ultrasonics ; methods

Comparison of pancreaticoduodenal transplants (PDT) and duct-ligated pancreas transplant (DLPT) were performed using syngeneic and allogeneic studies in rats. Both DLPT and PDT allogeneic grafts showed mild rejection. DLPT groups showed disorganized pathology and acini replaced by fat. Eventually, massive fibrosis was seen in the Islets of Langerhans, as well as rejection cellular infiltrates. In both PDT groups, normal histology was observed in the same period. Thus the effect of duct occlusion is highly detrimental for the grafts.
Animals ; Graft Rejection/pathology ; Ligation/adverse effects ; Pancreas/*pathology ; Pancreas Transplantation/*adverse effects ; Pancreatic Ducts/surgery ; Postoperative Period ; Rats ; Rats, Inbred Lew ; Rats, Sprague-Dawley ; Research Support, Non-U.S. Gov't ; Transplantation, Homologous ; Transplantation, Isogeneic

OBJECTIVETo screen specific functional proteins from lymphocyte involved in acute rejection using differential proteomics research.

METHODSTwo groups of rat liver transplantation models were established (isograft as control and allograft as acute rejection groups) by transplantation within Wistar rats, and between Wistar and SD. Morphology study were performed by histochemistry tech, followed by serum cytokine detection with ELISA. With 2-dimensional electrophoresis, proteomes of lymphocyte from the rats of different groups were separated and 2 proteome profiles were established. Comparing with the 2 profiles, 25 spots were selected and picked for in gel digestion, followed for analysis by matrix assisted laser desorption ionization (MALDI)-time of fly (TOF)/TOF MS. Two of the proteins were detected with Western blot to verify the changing profiles.

RESULTSThe results of morphology analysis and detection of cytokines (IL-2 and IFN-gamma) indicate that the animal models were established successfully and acute rejection happened after transplantation for 3 days. Twenty-five differential proteins were found out to be associated with acute rejection, among which 13 proteins were upregulated and 12 downregulated. The expression alterations of 2 proteins (beta-actin and carbonic anhydrase) are consistent with proteomics analysis results showing in Western blot.

CONCLUSIONSTwenty-five specific proteins exploiting mechanism of acute rejection are screened out, including IL-2 and carbonic anhydrase, which maybe benefit for the further works.

Acute Disease ; Animals ; Disease Models, Animal ; Graft Rejection ; metabolism ; pathology ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Liver ; pathology ; Liver Transplantation ; methods ; Lymphocytes ; metabolism ; Peptide Mapping ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transplantation, Homologous ; Transplantation, Isogeneic

In this study antigen-independent factor in the pathogenesis of chronic rejection of organ transplants was examined. Kidney isografts and allografts were transplanted orthotopically into bilaterally nephroectomized rat recipients and studied functionally, morphologically and immunohistologically, at serial intervals up to 52 weeks after transplantation. Allograft recipients developed progressive proteinuria after 12 weeks, with gradual renal failure ultimately leading to death. At the same time, morphological changes, including progressive arteriosclerosis and glomerulosclerosis, tubular atrophy and interstitial fibrosis, developed. Immunohistologically, macrophages infiltrated glomeruli during this period and cytokines became unregulated. Our results showed that antigen-independent functional and morphological changes occurred in long-term kidney isografts and mimicked those appearing much earlier in allografts that reject chronically. Initial injury and extent of functioning renal mass is suggested to be important factor for such late changes.
Animals ; Graft Rejection ; etiology ; immunology ; pathology ; Graft Survival ; physiology ; Kidney ; immunology ; pathology ; Kidney Transplantation ; immunology ; methods ; pathology ; Proteinuria ; etiology ; Rats ; Rats, Inbred Strains ; Time Factors ; Transplantation, Homologous ; Transplantation, Isogeneic