Objective To explore the potential applications of a chamber for in vivo tissue engineering,and to establish a novel model for in vivo tissue-engineered cartilage.Methods Auricular chondrocytes were isolated,cultured and identified from the ears cartilages of New Zealand white rab bits; rabbit auricular chondrocytes(RACs) were seeded into the scaffolds:(1) RACs were seeded into collagen gel scaffold; (2) RACs were seeded into PLGA/collagen gel scaffold in vitro,and the compos ites were placed into the chambers and implanted in the donor rabbit.As control groups,the composites were implanted directly subcutaneously in the donor rabbit without using chambers,and the contents were harvested at 8 weeks after implantation.Gross examination,histologic and immunohistochemical staining and RT-PCR test were performed to evaluate the harvested contents.Results Under the same conditions inside the chambers,the contents formed into new cartilage-like tissue by histo logical and immunohistochemical staining and RT-PCR.In contrast,in the control groups without chambers displayed vascular invasion and inflammatory reaction in the subcutaneous layer of skin,which eventually led to fibrous tissue or absorption.Conclusions Cartilage is successfully constructed in an immunocompetent animal model using a bioinert perforated chamber.This method is effective in creating a relatively favorable environment for cartilage regeneration,which may provide a valuable reference for the clinical application of tissue regeneration.

Objective To investigate the anti-arrhythmias mechanism of nardostachyos chinensis batal by intracardiac electrophysiologic study. Methods Intracardiac electrophysiologic study were performed in 40 supraventricular tachycardia(SVT)patients before and after oral nardostachys chinensis batal administraton,and the results were compared with 35 SVT patients or normal persons without nardostachys chineusis batal intake. Results The atrial effective,relative,functional refractory periods(AERP,ARRP,AFRP)and ventricular ERP(VERP)were significantly longer in nardostachyos chinensis batal administration group than control(AFRP(358 ± 76)ms vs.(256 ±69)ms;ARRP(365 ±74)ms vs.(287 ±55)ms;VERP(264 ± 36)ms vs.(213 ± 27)ms respectively);but the sinus and atrioventricular node function had no changes. Conclusions The anti-arrhythmia mechanism of nardostachyos chinensis batal were related to the prolonged cardiac refractory periods and the reduced cardiac excitability.

Objective:To construct the siRNA expression vector of focal adhesion kinase(FAK) gene and inhibit the expression of FAK gene in tongue cancer cell line Tca8113 by RNA interfering technique. Methods:According to the encoding sequence of FAK mRNA, two pieces of oligonucleotide sequences were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pGCSilencerTM-U6/Neo siRNA expression vector. After being identified by restriction enzyme method, the recombinant pSilencer-FAK plasmids were transfected into Tca8113 cells. The transfected cells were selected by G418 method. Immuocytochemistry and Western blotting were used to evaluate FAK gene silencing efficiency. Results:The oligonucleotide fragments were correctly inserted into pGCSilencerTM-U6/Neo vector. FAK expression of the transfected cells was significantly down-regulated by pSilencer-FAK. Conclusion:The siRNA expression vector of FAK is successfully constructed and FAK expression of Tca8113 cells can be inhibited by RNA interfering technique.

OBJECTIVETo observe the ultrastrual changes of the skin between the continuous tissue expansion and the conventional tissue expansion.

METHODSTwelve white piglets were used for this study on an animal model of tissue expansion with the continuous tissue expansion group and the conventional tissue expansion group. The tissue samples in each group were harvested and prepared for the transmission electron microscope observation.

RESULTSThe interspace among basal cells and spinose cells was increasing and the numbers of cell conjunctures were decreasing in the both groups. However, these changes in the continuous tissue expansion group were more obvious than in the conventional tissue expansion group. In the dermal layer of the skin, the ultrastructure of collagen fibers were basically normal. But, the fibroblasts and capillary endothelia cells were more activated in the continuous tissue expansion group, compared with the conventional tissue expansion group. The fibroblast apoptosis and collagenolysis spots were observed in both of the groups, while the red blood cells were also found in the tissue leaked outsides from the blood vascular cavities.

CONCLUSIONTissue expansion may result in tissue growth and tissue degeneration in the same time.

Animals ; Microscopy, Electron ; Skin ; cytology ; ultrastructure ; Swine ; Tissue Expansion ; methods

OBJECTIVETo investigate the contraction mechanism of expanded tissue.

METHODSWhite domestic pigs were employed to establish tissue expansion model. Tissue samples were taken after expansion for contraction measurements and histopathological observation.

RESULTSDermis and capsule were the main positions of contraction. In the dermal layer, elastic fibers and myofibroblasts proliferated greatly and alpha-Actin enhanced in the myoepidermal cells and capillary endothelial cells during expansion. In the capsule, the direction of elastic and collagen fibers arranged crossly and formed into special contractible structure.

CONCLUSIONElastic fibers, myofibroblasts and alpha-Actin enhancement of cells were the substantial materials of tissue contracture increases. The results suggest that suitable tension must be applied into expanded flaps for anti-contraction.

Animals ; Muscle Contraction ; Surgical Flaps ; Swine ; Tissue Expansion

Objective To compare the cumulative analgesic effects of electroacupuncture at Sanyinjiao (SP6), Xuanzhong (GB39) and non-acupoint in treating primary dysmenorrhea. Method By adopting a multi-centered randomized controlled study method, 501 patients recruited from Dongzhimen Hospital of Beijing University of Chinese Medicine, China-Japan Friendship Hospital, Beijing Hospital of Traditional Chinese Medicine of Capital Medical University, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Huguosi Hospital of Chinese Medicine of Beijing University of Chinese Medicine and the Outpatient of Shandong University of Traditional Chinese Medicine were randomized into a Sanyinjiao group, a Xuanzhong group, and a non-acupoint group, 167 subjects in each group. The electroacupuncture intervention was applied when dysmenorrhea flared up and the Visual Analogue Scale (VAS) ≥40 mm, with frequency at 2/100 Hz and intensity during patient’s endurance, 30 min each time, once a day, and for successive 3 d. Before the first treatment, 30 min after the first treatment, and respectively prior to the second and third treatment, VAS was used to measure the pain intensity. Meanwhile, the Retrospective Symptom Scale (RSS-COX 2) was investigated before the first treatment, right after the removal of needles for the first treatment, before the second and third treatment. Result The decrease of VAS in Sanyinjiao group was more significant than that in Xuanzhong group and non-acupoint group (MD=﹣2.92 mm, P=0.028; MD=﹣3.47 mm, P=0.009), while there was no significant difference between Xuanzhong group and non-acupoint group (MD=﹣0.56 mm, P=0.674); there were no significant differences in comparing the RSS-COX2 total score among the three groups (P=0.086). Conclusion Sanyinjiao (SP6) can produce a more significant cumulative analgesic effect for primary dysmenorrhea patient than Xuanzhong and non-acupoint, and the effects of Xuanzhong and non-acupoit are equivalent.

OBJECTIVETo investigate the expression and clinical significance of enhancer of zeste homolog 2 (EZH2) and p53 proteins in cervical squamous cell carcinoma (SCC) and cervical intraepithelial neoplasia (CIN).

METHODSThe expression and distribution of EZH2 and p53 were determined with reference to clinicopathological features and patient survival. 168 cervical SCC, 19 CINII, 35 CINIII patients and 30 normal control cases were collected for immunohistochemical analysis.

RESULTSThe expression of EZH2 in the normal cervix, CIN and SCC was 6.7% (2/30), 37.0% (20/54) and 75.6% (127/168), respectively, with a significant difference between them (P < 0.05). The expression of p53 was 3.3% (1/30), 20.4% (11/54) and 39.3% (66/168), respectively, also (P < 0.05). In the 168 SCC cases, the positive rate of EZH2 in the cases with lymph node metastasis was 82.9%, and that of p53 was 45.7%; the positive rate of EZH2/p53 protein expression in the cases with negative lymph nodes was 70.4%, and that of p53 was 34.7%, with a significant difference between the two subgroups (P < 0.05). Among the 143 followed-up SCC patients, the EZH2(+) p53(+) cases had a progression-free survival of (51.3 ± 3.8) months, that of EZH2(+) or p53(+) cases was (66.1 ± 2.0) months, and that of EZH2(-) p53(-) cases was (71.3 ± 1.9) months, with a very significant difference among the three subgroups (P < 0.001). The overall survival times of EZH2(-) p53(-), EZH2(+) or p53(+), and EZH(2+) p53(+) cases were (72.9 ± 1.1), (68.6 ± 1.8), and (57.4 ± 3.4) months, respectively, with a significant difference among the three subgroups (P < 0.001). The multivariate analysis showed that EZH2 expression, lymph node metastasis and tumor staging were independent prognostic factors.

CONCLUSIONSBoth EZH2 and p53 proteins may play important roles in the occurrence and development of cervical squamous cell carcinoma. There is a close relationship between the expression of both EZH2 and p53 proteins and the prognosis of SCC patients.

Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Cervical Intraepithelial Neoplasia ; diagnosis ; metabolism ; Disease-Free Survival ; Enhancer of Zeste Homolog 2 Protein ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Staging ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prognosis ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Uterine Cervical Neoplasms ; diagnosis ; metabolism