Purpose:To look for an ideal screening method in vitro for establishing osteosarcoma cell sublines with different metastat ic potential. Methods:20 osteosarcoma cell sublines were isolated preliminari ly by clone technique in vitro. They were screened by electrophores migratio n rate assay and cell migration assay in vitro and obtained respectively a h igh and low metastatic potential osteosarcoma cell sublines (A1、A2、B1 and B2). The advantages of the two methods were compared and confirmed by using cell pro liferation, agarose clony-formation and transplantation in nude mice. Results:The cell proliferation rate , agarose clony-formation ability and spontaneous metastatic ability to lung of A1 and B1 were obviously h igher than that of A2 and B2 and the difference was statistically significant( P

[Objective]To investigate the injury characteristics and treatment of the Hoffa facture associated with extensor mechanism injury.[Method]Four patients with Hoffa facture associated with extensor mechanism injury treated in author's hospital in recent 6 years were enrolled in this study.All the patients were multiple-injured,and associated with the ipsilateral limb fracture and retinacular and capsule laceration.The site of the extensor mechanism injury included 1 distal patellar tendon disruption,2 proximal patellar tendon disruption,and 1 distal quadriceps tendon disruption.No cruciate ligament injury was found in 3 ipsilateral bicondylar fractures.One unicondylar fracture was associated with anterior and posterior cruciate ligament and medial collateral ligament injury simultaneously.Emergency operation was performed along with the debridement in the three open fractures.In another patient with closed fracture,operation was performed one week after injury.Fractures were reduced and fixed with lag screws.Extensor apparatus were repaired and strengthened with a tension band wire through both patella and tibia.Ipsilateral limb fractures were reduced and internal fixed with hardware simultaneously. The knees were immobilized in extension with cast after operation,and flexion exercise was commenced four to six weeks later.[Result]Four patients wounds reached the first rate healing.There was no infection and necrosis of skin.All patients were followed-up for six months to three years.All 7 condyles of the 4 patients reached bone union.Healing time was about six to twelve weeks.According to Letenneur’s functional assessment system,excellent and good results were in 1 cases,fine in 3 cases.[Conclusion]Hoffa facture associated with extensor mechanism injury are usually accompanied with injuries in other part of the body.In open fracture,emergency operation is mandatory,and fracture reduction and internal fixation and extensor apparatus repair should be performed along with debridement.Tension band wire may strengthen the stability of the extensor mechanism and facilitated early mobilization.

To determine whether increased expressions of gelatinase A(MMP-2) and gelatinase B(MMP-9) occur in vivo in autoimmune MRL/lpr mice model and to investigate the modulation effects of "shenle." Shenle (4g?kg -1 ?d -1 ,orally) or methylprednisolone(MPS,25mg?kg -1 ?d -1 ,ip)was administered daily to MRL/lpr mice at the age of 8 weeks. The activities of MMP-2/9 by gelatin zymography were compared in kidney protein extracts and urine. After treatment for 20 weeks, a progressive reduction in positive proteinuria number/total mice (40% vs.33 3% vs.80%, proteinuria over 300 mg/dl as positive) and an elevated survival rate (70% vs.80% vs. 50%) were found in "shenle" and MPS groups compared with the control group. Histological analysis of kidney tissues indicated that both "shenle" and MPS could inhibit the mesangial proliferation and renal sclerosis. Using SDS-PAGE gelatin zymography, we have identified increased expressions of both latent and activated form enzymes of MMP-2/9 in urine and kidney extraction. Immunohistochemical staining showed both MMP-2 and MMP-9 were obviously up-regulated within glomerulus in control group. "Shenle" as well as MPS suppressed the expression of both latent and activated form of MMP-2/9. These in vivo results suggested that MMP-2/9 expressions might play an important role in murine lupus nephritis. "Shenle" delayed the development of glomerulonephritis and improved survival in MRL/lpr mice probably by suppressing the expressions and activities of MMP-2/9.

To determine thrombin activation and fibrin deposition in the development of lupus nephritis in MRL lpr/lpr mice and the inhibitory effects of "shenle". "Shenle" (4g/(kg?d) orally) was administered daily to MRL lpr/lpr mice at the age of 8 weeks. After treatment for 20 weeks, we compared thrombin receptor (Protease Activated Receptor-1, PAR-1) expression with immunohistochemistry and fibrin deposition with MSB(Martius-Scarlet-Blue)staining in renal sections. PAR-1 mRNA expression was analyzed with RT-PCR method in the two groups. With the development of murine lupus nephritis, we observed an increase in thrombin receptor mRNA and severe fibrin deposition in renal tissue in the control group, while thrombin receptor protein expression was strikingly downregulated, suggesting its continuous activation and degradation. "Shenle" inhibited PAR-1 activation significantly and it was correlated with reduced fibrin deposition. These results suggested that thrombin activation may play an important role in the development of glomerulonephritis in MRL-lpr mice. "Shenle" ameliorated the murine renal lesions probably by inhibiting thrombin receptor activation and fibrin deposition.

Objective To evaluate the condition of living renal donors with 3-dimensional volume rendering computed tomography (3D-CT) in preoperation. Methods 10 consecutive cases received hand-assisted transperitoneal laparoscopic living donor nephrectomy (HLDN) from December 2001 to July. Total patients underwent 3-dimensional volume rendering computed tomography before operation for assessing the condition of the anatomical location, number, anomalies and spatial interrelationships of the renal arteries and veins. These 3D-CT were directly compare with intraoperative surgical findings at laparoscopy and the harvested kidney ex vivo to value the sensitivity and accuracy of 3D-CT. Results The CT digital data of 10 cases were reconstructed to 3D-CT with volume rendering technique, and HLDN were performed successfully in all the cases. The 3D-CT accurately identifies renal vascular anatomy. Conclusions Not only the 3D-CT accurately identifies renal vascular anatomy but also tridimensional images reconstructed by the enhanced CT digital data of arterial phase, venous phase and renal parenchymal phase are viewed by surgeon in any position. It is a manner that may facilitate renal hilar dissection during laparoscopic donor nephrectomy. The 3D-CT should be a routine examination for living donor in preoperation

Objective To study the chimerism formation in kidney transplantation recipient receiving peripheral blood stem cells (PBSCs) of the donor after the patient received preoperative total lymphoid irradiation (TLI). Methods 5 patients of living donor kidney transplantation were involved in present study. The mean age of donors was 40 and that of recipients was 27. There was one patients with 4 HLA antigen mismatches out of 6 HLA-A, B, DR antigens, two patients with 3 HLA-mismatches, and two patients with 1 HLA-mismatch. The patients received TLI in doses 90cGy/d from day -5 to day -1. The immunosuppression protocol for these patients during operation and afteroperation was similar to that for other cases, but the dosage was slightly reduced. Donor PBSCs were harvested twice via leukapheresis after the administration of human recombinant granulocyte colony stimulating factor (G-CSF). PBSCs were infused intravenously to the recipients on postoperative day 4 and 7. Chimerism of peripheral blood cell in recipients was detected by PCR-SSP assays. The grafted kidney function, acute rejection frequency and GVHD episodes were also observed. Results After 5 days of G-CSF based mobilizing regimen, the average number of PBSCs harvested from donors was 34?10~6. At 1 month post-transplantation, donor type HLA-DR gene was detected in all the recipients. At 2 and 3 months post-transplantation, chimerism was not detectable in the recipient with 4 HLA antigens (including 2 HLA-DR) mismatched donor kidney and in the recipient with 3 HLA (including 2 HLA-DR and 1 HLA-B) mismatched donor, respectively. In the other 3 recipients, chimerism was still positive 3 months post-transplantation. There were no symptoms of GVHD or infections after infusion of PBSCs. Transplanted kidneys functioned normally and no rejection episodes were observed till the end of the study. The immunosuppressant dosage was lower for these 5 cases than those after conventional cadavar renal transplantation. Conclusions Non-myeloablative TLI conditioning regimen is a safe and effective method which can promote the engraftment of donor PBSCs and induce hemopoietic chimerism. Chimerism can alleviate rejection of the transplanted kidney and reduce the immunosuppressant dosage. Collection and infusion of PBSCs are simple, convenient, effective, and inexpensive, and it is an optimal source of hemopoietic stem cells.

Objective To determine the changes in expressions of thrombin receptor and fibrin deposit in glomeruli during the process of senility. Method Rats were divided into 3 groups (8 rats in each group): 3-month-old group (3m), 12-month-old group (12m) and 24-month-old group (24m). Fibrin deposition was detected by Martius-Scarlet-Blue staining and direct immunofluorecence method. Immunohistochemical studies were performed to detect the expression of thrombin receptor (PAR-1) and transforming growth factor-? (TGF-?). Semi-quantitative PCR was performed to detect the changes in PAR-1 mRNA expression. A quantitative analysis of the expressions was performed by image analysis system. Result Significant pathological changes were found in glomeruli during the process of senility. Fibrin deposition was not observed in glomeruli in different groups. Significant expression of PAR-1 was found in glomerular endothelial cells, mesangial cells and epithelial cells in 3m rats. On the contrary, in 24m rats, PAR-1 expression in glomeruli was significantly decreased. Expression of TGF-? was increased with senility in glomeruli. PAR-1 gene expression, barely detectable in control tissue, was strikingly increased in 24m rats. Conclusion Thrombin receptor activation could be found in glomeruli of senile rat, and it is independent of fibrin deposition. Activation of PAR-1 may play an important role in the process of renal senility.

Objective To investigate the safety and effectiveness of the application of an immunosuppressing primed total lymphoid irradiation (TLI) of the recipient in kidney transplantation of living donor kidney graft. Methods 5 recipients, 3 males and 2 females with the average age of 27, underwent the primed regimen of TLI. As to 6 HLA-A, B, DR antigens of donors and recipients, 4 HLA antigen mismatches were found in one case, 3 mismatches in two cases, and 1 mismatch in two cases. The donor grafts were procured by the technique of hand-assisted transperitoneal laparoscopic living donor nephrectomy (HLDN). The primed regimen of TLI was carried out 5 days before the operation in a dosage 90 cGy/d. The intra- and postoperative immunosuppressant protocols for these cases were similar to that for the other cases, but the doses were reduced slightly. WBC, total T cell, CD4~+ and CD8~+ T cell subsets and Th1/Th2 immune deviation were examined in recipients periodically. The function of the grafted kidney, acute rejection frequency, and side effects of TLI were also observed. Results In the recipients undergone TLI, the total WBC and T cell, CD4~+ and CD8~+ subsets were reduced to a base value at 1-2 weeks post-transplantation and did not restore to the pre-transplantation levels until the investigation was ended. A particular phenomenon showing that in TLI based regimen T cells were shifted from Th1 to Th2 cells was observed. No side effects of TLI, such as infection, were found during the observation. Transplanted kidneys functioned normally and no rejection episodes were observed up to the end of the study. The immunosuppressant doses for those 5 cases were lower than that of conventional protocol. Conclusions TLI based non-myeloablative primed regimen is a safe and effective method for immunosuppression without serious side effects, and it can markedly alleviate kidney transplant rejection and reduce the dosage of immunosuppressive drugs.

Objective To investigate the clinical value of multi-slice spiral CT (MSCT)in diagnosis of internal abdominal hernia (IAH).Methods The MSCT findings in 1 5 patients with IAH confirmed by surgery were retrospectively analyzed.Results MSCT showed intestinal obstruction in 14 patients with strangulating intestinal obstruction in 8.The typical features of MSCT were as follows:translocated position of intestines (occupying effect),closed loop intestinal obstruction (gathered intestinal loops with different forms),assembled,expanded and translocated blood vessels of mesenteries,and thickened intestinal walls with ischemic edema and abnormal density.Surgical results showed paraduodenal hernia in 2,transomental hernia in 2,transmesenteric hernia in 5,ankylenteron strap hernias in 4,pericecal hernia in 1 and hernia through the Douglas fossa in 1,Intestinal obstruction in 14 and strangulating intestinal obstruction in 8 were also confirmed by surgery.The CT findings of IAH were correlated with the surgical results,and the accurate rate of grading of intestinal obstruction with CT was also consistent with the surgical results (Kappa=0.758,t=3.462,P=0.001).Conclusion IAH can be diagnosed accurately by MSCT in most patients,MSCT is helpful for guiding the surgical programs.

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