Objective To investigate the protective effect of total flavonoids of drynaria on hemorrhagic shock reperfusion renal injury.Methods The 30 Sprague-Dawley rats were randomly divided into 3 groups: sham operation group, experimental group (hemorrhagic shock group) and drynaria fortunei group, 10 in each group. In drynaria fortunei group and experiment group, the blood from the femoral artery was with drawn until the mean arterial pressure reached 40 mmHg in 90 min, and then the withdrawn blood transfusion were reinfused and blood pressure recovered to the base value of 90%. The rats of drynaria fortunei group were given intraperitoneal injection of Drynaria total flavonoid 300 mg/kg, and experimental group and sham operation group were injected with 3 ml physiological saline. Blood creatinine (SCr) levels, the renal cell apoptosis index, the expression of Bcl-2 and mRNA Bax in kidney were detected. Results Compared with the experimental group, serum SCr (78.36 ± 5.31 mmol/Lvs. 151.43 ± 11.8 mmol/L) of drynaria fortunei group decreased significantly (P<0.01); the renal cell apoptosis (19.56% ± 4.37%vs. 14.31% ± 2.83%), the expression level of Osteopractic rats of Bax group mRNA (1.24 ± 0.05vs. 0.91 ± 0.07) of drynaria fortunei group decreased significantly; and the Bcl-2 mRNA (0.65 ± 0.05vs. 1.38 ± 0.07) and the expression level and Bcl-2/bax ratio (0.52 ± 0.06vs. 1.51 ± 0.08) of drynaria fortunei group increased significantly (P<0.01).Conclusions The drynaria total flavonoids on rats with hemorrhagic shock and resuscitation of renal injury has a protective effect, and its mechanism may improve the expression of Bcl-2 and mRNA, increase the ratio of bcl-2/bax, and inhibit  or block the apoptosis of renal tissue.

Objective To evaluate the feasibility of reconstruction of artificial bladder tissue using the expanded bladder cells seeded onto both sides of a synthetic PLGA polymer and cultured in the nude mice by tissue engineering technique. Methods The urothelial cells and smooth muscle cells were obtained from young rabbits by mechanical isolation and enzyme digestion method. In vitro cultured and expanded smooth muscle cells and urothelial cells were seeded onto the outer and inner surfaces of each polymer. Then, the constructs were implanted into the subcutaneous pockets of athymic mice. At 4 and 8 weeks after implantation, the specimens were harvested and examined macroscopically, histologically, and immunohistochemically. Results The polymer was covered with urothelial cells inside and smooth muscle cells outside. As polymer degradation was in progress, the urothelial cells and the smooth muscle cells kept proliferation and converged. Conclusion Artificial bladder tissue reconstructed by tissue engineering technique has the similar properties to those of the normal bladder wall. This study has laid solid foundation for further studies of tissue engineering bladder.

AIM: To set up a method of inducing mouse embryonic stem cells (mESC) to differentiate into cardiomyocyte after treatment with 5-azacytidine. METHODS: Cytotoxicity of 5-azacytidine was measured by MTT assay. Treatment of mESC with conditioned culture mediums, which were composed of 5-azacytidine alone or combined with retinoic acid, induced the cell differentiation to cardiomyocytes. The cells induced were identified by detecting the expression of cardiac proteins (myosin, desmin, ?-actin and ?-actinin). Gene MLC-2v, a specific gene of ventricular-like cardiomyocyte, was also detected by RT-PCR. RESULTS: The non-cytotoxic dose of 5-azacytidine was 8 ?mol/L, which was able to induce mESC to differentiate into cardiac syncytiums. Cells induced expressed many cardiac proteins and MLC-2v mRNA. However, combined with retinoic acid inhibited mESC differentiation into cardiomyocyte. CONCLUSION: 5-azacytidine is able to promote mESC differentiation into cardiomyocytes. A method of inducing mESC to differentiate into cardiomyocytes in vitro has been established.

Objective To probe the periodontal ligament regeneration following the implantation of bone marrow mesenchymal cells ( BMSCs ) sheet-collagen membrane-BMSCs sheet sandwich complex.Methods BMSCs cell sheet-collagen membrane-BMSCs sheet complexes were compounded on the root surface of teeth of Beagle dogs.All the dogs were killed on 4 and 12 weeks after implantation.Periodontal ligament regeneration was observed by radiological means, HE staining and Sirus-red staining.Results Compared with collagen membrane group and blank control group, there was a clearly periodontal ligament like tissue and Sharpey′s like fibres formation in test group only.Conclustion Cell sheet-collagen membrane-cell sheet sandwich complex can effectively improve the periodontal ligament regeneration.

In order to study the chondrogenic phenotype differentiation of adult sheep bone marrow-derived mesenchymal stem cells (MSCs) in a defined medium as potential seed cells for cartilage tissue engineering. MSCs were isolated by density centrifugation with Percoll solution from bone marrow aspirated from sheep iliac crest. The third passage of MSCs were induced with H-DMEM containing TGF-beta3, IGF-I, Dexamethasone and VitC. The shape and ultrastructure of cells were observed, toluidine blue stain for GAG and immunohistochemistry for type II collagen were applied for chondrogenic phenotype identification. After 14 days of induction, MSCs changed from a spindle-like appearance to a polynal shape, a large amount of endoplasmic reticulum, Golgi complex and mitochondria were observed, and the differentiation of MSCs chondrogenic phenotype was verified by positive staining of toluidine blue and immunohistochemistry. MSCs derived from bone marrow can differentiate to chondrogenic phenotype when induced in vitro and can be used as optimal seed cells for cartilage tissue engineering.
Bone Marrow Cells/*cytology ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Chondrocytes/*cytology ; *Chondrogenesis ; Mesenchymal Stem Cells/*cytology ; Phenotype ; Sheep ; Tissue Engineering

BACKGROUND AND OBJECTIVERadiation sensitivity is closely related to tissue oxygen, and rh-endostatin can induce the high level of oxygen content in tumor by "normalizing" tumor angiogenesis which is associated with radiotherapy sensitivity. The aim of this study is to observe the effect of combination of radiotherapy with rh-endostatin in the rats with lung cancer.

METHODSImmediate lewis cancerous ascetic injection method was used to make rats tumors bearing model, then the rats was divided into four groups randomly: group A was treated with saline; group B was treated with rh-endostatin; group C was treated with irradiation and group D was treated with rh-endostatin and irradiation. After all rats were treated, inhibition rates and the tumor growth curve were calculated. Immunohistochemisty was adopted to check the expressions of vascular endothelial growth factor (VEGF) and microvessel density (MVD).

RESULTSCompared with group A, the growth rates of the tumors in the other group were obviously slower, and the tumor weights were significantly different form group A (P < 0.05). Compared with the other groups, the tumor weights of group D were obviously reduced (P < 0.05). Compared with group A, VEGF and MVD of other three groups were reduced (P < 0.05), and group D were significantly cut down.

CONCLUSIONCombination with radiotherapy and rh-endostatin could inhibit the lung cancer significantly in rats. The possible mechanisms are to decrease the expression ofVEGF and inhibit the production of angiogenesis.

Animals ; Carcinoma, Lewis Lung ; drug therapy ; metabolism ; pathology ; radiotherapy ; Endostatins ; therapeutic use ; Female ; Immunohistochemistry ; Lung Neoplasms ; drug therapy ; metabolism ; radiotherapy ; Mice ; Mice, Inbred C57BL ; Microvessels ; pathology ; Random Allocation ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo evaluate the feasibility of growing tissue-engineered cartilage using chondrocytes seeded onto a biodegradable porous bioceramic, the beta-tricalcium phosphate (beta-TCP).

METHODSA porous bioceramic template of beta-TCP was created in the shape of a disc. Chondrocytes isolated from rabbit articular cartilage were seeded on the beta-TCP template and then kept in rotatory cell culture system (RCCS) for 1 week prior to subcutaneous transplantation into athymic mice. The three-dimensional structure was well-maintained 16 weeks after implantation. After 4, 8, 16 weeks, the specimens were harvested and examined macroscopically, histologically and immunohistochemically.

RESULTSGross morphological and histological analysis of the specimens from the chondrocyte-beta-TCP complex demonstrated new cartilage construction. The overall configuration of the experimental specimens closely resembled the structure of beta-TCP template.

CONCLUSIONThese findings suggest that porous bioceramic (beta-TCP) is a good "matrix" for chondrocyte, and can be used for cartilage engineering.

Animals ; Calcium Phosphates ; pharmacology ; Cartilage ; growth & development ; transplantation ; DNA ; analysis ; Female ; Glycosaminoglycans ; analysis ; Immunohistochemistry ; Mice ; Mice, Nude ; Tissue Engineering

In order to study the chondrogenic phenotype differentiation of adult sheep bone marrow-derived mesenchymal stem cells (MSCs) in a defined medium as potential seed cells for cartilage tissue engineering. MSCs were isolated by density centrifugation with Percoll solution from bone marrow aspirated from sheep iliac crest. The third passage of MSCs were induced with H-DMEM containing TGF-beta3, IGF-I, Dexamethasone and VitC. The shape and ultrastructure of cells were observed, toluidine blue stain for GAG and immunohistochemistry for type II collagen were applied for chondrogenic phenotype identification. After 14 days of induction, MSCs changed from a spindle-like appearance to a polynal shape, a large amount of endoplasmic reticulum, Golgi complex and mitochondria were observed, and the differentiation of MSCs chondrogenic phenotype was verified by positive staining of toluidine blue and immunohistochemistry. MSCs derived from bone marrow can differentiate to chondrogenic phenotype when induced in vitro and can be used as optimal seed cells for cartilage tissue engineering.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Chondrocytes ; cytology ; Chondrogenesis ; Mesenchymal Stromal Cells ; cytology ; Phenotype ; Sheep ; Tissue Engineering