The feasibility of bone marrow stromal cells autologous transplantation for rabbit model of dilated cardiomyopathy induced by adriamycin was studied. Twenty rabbits received 2 mg/kg of adriamycin intravenously once a week for 8 weeks (total dose, 16 mg/kg) to induce the cardiomyopathy model with the monitoring of cardiac function by transthoracic echocardiography. Marrow stromal cells were isolated from cell-transplanted group rabbits and were culture-expanded on the 8th week. On the 10th week, cells were labeled with 4,6-diamidino-2-phenylindole (DAPI), and then injected into the myocardium of the same rabbits. The results showed that viable cells labeled with DAPI could be identified in myocardium at 2nd week after transplantation. Histological findings showed the injury of the myocardium around the injection site was relieved with less apoptosis and more expression of bcl-2. The echocardiography found the improvement of local tissue movement from (2.12±0.51) cm/s to (3.81±0.47) cm/s (P<0.05) around the inject site, but no improvement of heart function as whole. It was concluded bone marrow stromal cells transplantation for dilated cardiomyopathy was feasibe. The management of cells in vitro, the quantity and the pattern of the cells transplantation and the action mechanism still need further research.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

In order to investigate the origin of neointimal smooth muscle cells in transplant arteriosclerosis in rat aortic allograft, sex-mismatched bone marrow transplantation was performed from male Wistar rats to female Wistar rats. Four weeks after transplantation, the aortic transplant model was established by means of micro-surgery in rats. The recipients were divided into 4 groups: female Wistar-female Wistar aortic isografts, female SD female Wistar aortic allografts, male SD-male Wistar aortic allografts, female SD-chimera Wistar aortic allografts. Eight weeks after transplantation, aortic grafts were removed at autopsy and processed for histological evaluation and immunohistochemistry. The results indicated that excessive accumulation of α-SMA-positive smooth muscle cells resulted in significant neointima formation and vascular lumen stricture in rat aortic allografts. Neointima assay revealed that the neointimal area and NIA/MA ratio of transplanted artery were significantly increased in all of aortic allograft groups as compared with those in aortic isograft group (P<0.01). Neointimal smooth muscle cells were harvested from cryostat sections of aortic allograft by microdissection method. The Sry gene-specific PCR was performed, and the result showed that a distinct DNA band of 225 bp emerged in the male-male aortic allograft group and chimera aortic allograft group respectively, but not in the female-female aortic allograft group. It was suggested that recipient bone-marrow cells, as the origin of neointimal smooth muscle cells, contributed to the pathological neointimal hyperplasia of aortic allograft and transplant arteriosclerosis.

To examine the expression of human telomere reverse transcriptase (hTERT), p53 and proliferating cell nuclear antigen (PCNA) in cystitis glandularis, 38 patients were divided into two grouips: group A (including 18 cases of papillary cystitis glandularis) and group B (including 20 subjects with normal bladder mucosa). All the cases were immunohistochemically examined by using antibodies specifically against p53 and PCNA, and hTERT was determined by in situ hybridization. hTERT was found in 6 cases (33.3%) and p53 was detected in 4 cases (22.2%) in group A, while they were not detected in group B. There were significant differences in hTERT and p53 expression between groups A and B (P<0.05 for both). PCNA was detected in 7 cases (38.9%) in group A and 1 case (5.0%) in group B, and significant difference in PCNA expression was found between the two groups (P<0.05). The expressions of hTERT, p53 and PCNA were significantly higher in group A than in group B, suggesting that papillary cystitis glandularis is predisposed to cancerous change, and p53, PCNA, hTERT may be related to the malignant alteration.

The effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) and osteogenic revulsants alone or in combination at different time points and in different dosages on proliferation and osteogenesis of bone marrow stromal cells (BMSCs) in SD rats were investigated. Rat BMSCs were cultured in vitro and induced by rhBMP-2 in different dosages (10, 50, 100 and 200 μg/L) alone or in combination with osteogenic revulsants. MTT colorimetric assay was used to evaluate The proliferation, activity of alkaline phosphoric (ALP) and osteocalcin were measured at 3rd, 6th, 9th, 12th day respectively. The results showed that rhBMP-2 and osteogenic revulsants could promote the differentiation of BMSCs towards osteoblast phenotype. The proliferation of BMSCs could be enhanced by rhBMP-2 in a dose-dependent manner. The expression of osteoblast phenotype was significantly higher by using both of them than by using them alone, which was verified by the activity of ALP and osteocalcin. It was suggested that the combined use of rhBMP-2 and osteogenic revulsants could promote the proliferation and simultaneously induce and maintain the expression of osteoblast phenotype of BMSCs in rats.

To explore the clinical value of contrast-enhanced ultrasound (CEUS) in differentiating benign and malignant focal liver lesions (FLLs) with SonoVue, CEUS was used to examine 113 patients with focal liver lesions (FLLs) in our hospital during July 2005 to December 2006. All the patients underwent contrast-enhanced CT (CECT) or contrast-enhanced MRI (CEMRI). Except for patients with focal fatty sparings (n=18) and with hemangiomas (n=8), all the patients were confirmed by operation or ultrasonic-guided liver puncture biopsy. A sulfur hexafluoride gas-based contrast agent was used with a MI of 0.15 to 0.17. Forty-eight cases of malignant FLLs, including 30 hepatocellular carcinomas (HCCs), 2 cholangiocarcinomas and 16 metastatic tumors, were detected. Seventy-eight cases of benign FLLs, including 33 hemangiomas, 9 focal nodular hyperplasias (FNHs), 19 focal fatty sparings, 5 abscesses, 7 regenerative nodules and 2 inflammatory pseudo-tumor, were involved. The contrast pattern of benign and malignant FLLs was quite different. CEUS has higher specificity and sensitivity than conventional ultrasound in differentiating benign and malignant FLLs.