Objective To screen microsatellite DNA markers from genome of guinea pigs for further genetic quality control and gene-mapping of this species .Methods Microsatellite sequences were obtained by magnetic bead enrichment and genome database screening , and candidate loci were chosen to design primers .Thereafter , genomic DNA of 5 different guinea pig strains were employed to select polymorphic microsatellite DNA markers based on PCR amplification results .Re-sults A total of 304 microsatellite sequences were analyzed by magnetic bead enrichment and 125 primers were designed . One polymorphic microsatellite DNA marker and 17 specific sites ( no polymorphic was found ) were determined .By gene-mapping , 292 microsatellite sequences were obtained and 178 primers were analyzed , totally 25 polymorphic microsatellite DNA markers and 28 specific sites ( without polymorphics ) were discovered .Conclusions We obtained 26 polymorphic microsatellite DNA markers and 45 potential markers in guinea pigs , and these may lay a foundation for application of mic-rosatellite DNA markers in genetic quality control and gene-mapping of guinea pigs .

Objective To investigate the changes at the motor end-plate (i. e. neuromuscular junction) following the free neurovascular muscle transfer. Methods Light and electron microscopy were used to study serially during 30 weeks the changes at the neuromuscular junction after free muscle transfer of the gracilis muscle in the adult Wistar rat. Results Under light microscopy, after staining with acetylthocholine the neuromuscular junction showed changes of degeneration with withdrawal of the innervating axon terminal followed by regeneration and reconstitution of the neuromuscular junction. With the electron microscope, mitochondrial sweelling and clumping of the synaptic vesicles were followed by withdrawal of the axon terminal from the muscle membrane on denervation. The infolding of the muscle membrane at the neuromuscular junction became less prominent. With reinnervation the ulatrastructure of the junction was only partially reestablishied with poorly reconstituted primary and secondary folds of the muscle membrane. The newly formed neuromuscular junction still lacked the structural detail seen in the control neuromuscular junction, even after 30 weeks. Conclusion The persistent weakness following free neurovascular muscle transfer may be attributed to the failure of full recovery of the intricate ultrastructure of the neuromuscular junction.

Objective To investigate the technique and method of MSCT in examination of laryngeal. Methods MSCT data of 28 patients with laryngeal carcinoma confirmed pathologically were analyzed retrospectively. Images quality was evaluated and the results obtained with various windows and CT virtual larynscopy were compared. Results All images had good quality,20 cases(69% ) could be displayed with conventional soft tissue windows,24 cases(81% ) could be demonstrated with lung windows,and 27 cases(93% ) could be demonstrated with CT virtual larynscopy. Conclusion The multislice CT could effectively demonstrate laryngeal carcinoma, and could be applied routinely in examination of laryngeal carcinoma.

PurposeTo investigate the image quality of Dual Source CT (DSCT) high pitch prospective scan in acute chest pain examination by using triple-rule-out (TRO) protocol and its responses to heart beat rate change.Materials and Methods Thirty-two consecutive patients who planned to undergo TRO acute chest pain examination were enrolled and randomly divided into research group and control group. The conventional retrospective spiral scanning protocol was applied to the patients in the control group, while the high pitch prospective spiral scanning mode was used in the research group. CT value, noise level, SNR, CNR and radiation dosage in aorta and pulmonary arteries were recorded and compared between the two groups, and the image quality of the coronary arteries was scored subjectively.Results No significant difference was found in the aspects such as CT value, noise level, SNR, CNR radiation dosage in aorta and pulmonary arteries (P>0.05); however, the subjective scoring for coronary artery in the research group (85.5%) was significantly lower than that in the control group (93.9%) (χ2=8.33,P<0.05). Considering the influence of severe heart rate changes, we excluded those patients whose heart rate variance was more than 5 beats/min (control group:n=5; research group:n=4) and then compared the image quality of the two groups again. The subsequent results showed no difference (χ2=0.12,P>0.05). In addition, the radiation dosage in the control group was significantly higher than that in the research group [(20.8±6.2) mSvvs. (2.3±0.4) mSv;t=17.142,P<0.01].Conclusion DSCT high pitch prospective scanning in TRO protocol shows high success rate in examining patients with acute chest pain whose heart beat rate is lower than 80 beats/min and heart beat variance is less than 5 beats/min. The image quality has no difference with that derived from retrospective scan but with far more less radiation dose (2.3±0.4) mSv. This method thus is promising and can be widely promoted clinically.

Objective To discuss the clinical value of application of three-dimensional reconstruction technique of MSCT in examination of colonic tumours.Methods Thirty patients underwent scanning using helical CT and post-processing on the workstation.All CTC images were reconstructed using shaded surface display (SSD) on workstation.Perspestive images such as the ones from DCBE were generated via raysum.The images could clearly demonstrate the extent and detail of the disorder by adjusting the CT value threshold.These results and that of barium enema and colon endoscopy were compared.Results Thirty cases had completely clinical and pathological data.3D images could correctly reveal the macroscopic morphology of colon tract and location of lesions.The correctly diagnostic rate was 100％ by MSCT,while the diagnostic rate by CTVE and colon endoscopy was basicly identical.Conclusion With the utilization of multiple methods,3D reconstruction can effectively demonstrate colonic tumours,and can be applied routinely in examination of colonic tumours.

Objective:To establish an experimental phalloplasty model with sensory restoration in rabbits and to explore a new surgical technique to improve the postoperative sensory function in phallic reconstruction. Methods: Adult male New Zealand white rabbit was used. Penile reconstruction was performed with superficial epigastric faciovascular pedicle flap. Free saphenous nerve graft was dissected, embedded in the flap, and end-to-end anastomosed to dorsal nerve of penis. Postoperatively, H-E staining and CGRP immunohistochemical staining were applied to observe the morphology of the regenerated nerves in the reconstructed penis at different stages. Results:Only a few CGRP-positive fibers scattered in deep corium in the reconstructed penis 1 month after surgery, while the quantity of CGRP nerve fibers increased markedly and the CGRP-positive fibers appeared in both superficial corium and papillae corii after 3 months. Six months postoperatively, the positive nerve fibers were also observed around the cutaneous accessories and in the epidermis-corium boundary layer. Conclusion: This rabbit model demonstrates the possibility of a new method for phalloplasty with sensory recovery, which may redound to clinical application.

ObjectiveTo study the inhibiting effects on the invasion and metastasis of melanoma by CXCR4 gene silence in nude mice.MethodsThe CXCR4 specific recombinant plasmid vector was constructed and transfected into the cultured MV3 cell line with lipofectamine.The models of subcutaneous melanoma in nude mice were established with MV3 cells.The nude mouse model of lung metastasis was established by injection of MV3 cells into the tail vein.The animals were sacrificed at 8weeks after the melanoma cells injection.CXCR4-shRNA plasmid vectors were discontinuously injected directly into the established tumor and vein.The changes of weight and size of the tumors and the mice body weight during the therapy were calculated respectively.Histological observation was performed to evaluate the presence and number of metastatic tumors.ResultsThe subcutaneous melanoma tumors in nude mice were established successfully.The growth of tumors in the CXCR4-shRNA injected nude mice was inhibitted obviously through tumor growth curve. There were significant differences between negative shRNA control nude mice and blank control nude mice (P＜0.01).Melanoma cells with CXCR4 shRNA permanent transfection had a much lower lung and brain and liver metastatic potential in nude mice than control cells and mock control cells in vivo.ConclusionsCXCR4 gene silencing mediated by shRNA significantly suppresses the growth of MV3 cell in vitro.Silencing of CXCR4 mediated by shRNA can also effectively decrease the metastatic potential of lung and liver and brain.

Objective To investigate the relationship between thyroid nodules with calcification and thyroid carcinoma and its significance in the screening of thyroid carcinoma in high risk group.Methods The clinical data of 1771 patient undergoing surgery for thyroid nodules from March, 2006 to March, 2009 in Huashan Hospital, Fudan University were retrospectively analyzed. Results Among 1771 patients, 500 were finally identified as having malignant tumors. Incidence of calcification in thyroid carcinoma was 68. 4%, and that in benign thyroid nodules was 27.0% ( χ2 = 259. 5, P ＜ 0. 05 ). The specificity of microcalcification for the diagnosis of carcinoma was 89. 4%, and its positive predictive value was 66. 3% ( χ2 = 368.6, P ＜ 0. 01 ). The incidence of thyroid carcinoma in patients ＜ 45 years was 39.2%, while that in patients ≥ 45 years was 22.9% ( χ2 = 51.12, P ＜ 0. 05 ). The incidence of carcinoma in patients of single thyroid nodule was 31.7% and that in those with multiple nodules was 26. 4% (χ2 =4. 766,P ＜ 0. 05). Metastasis was pathologically diagnosed in 26. 8% of lymph nodes found by preoperative ultrasonography. Conclusions The specificity of thyroid nodule calcification, especially microcalcification is high for the diagnosis of thyroid carcinoma. High-risk index for carcinoma includes thyroid nodules with microcalcification, ＜ 45 years old and single thyroid nodule.

Objective To study the role of bone morphogenetic protein-7 in the osteogenic differentiation of periosteal cellsin vitro. Methods Periosteal cells, obtained from adult tibial periosteum, were cultured by routine method in vitro, and divided into two groups. One group cultured with BMP7 and the supplements of 100 nmol dexametasone, 10 mmol b-glycerophosphate and 50 mg/mL L-ascorbic acid (BMP7 group), the other cultured with the supplements alone as the control (control group). Ultrastructure and morphological changes of periosteal cells were observed by contrast phase microscope and electron microscope. In order to test the expression of markers of osteoblastic differantiation in periosteal cells, involved mineralized node and alkaline phosphatase. Each group was tested at the time of 5 d, 10 d, 15 d, 20 d, respectively, using ALP kit stain and Von Kossa stain with 3 samples at each time. Results The periosteal cells cultured by routine method and induced into osteoblast differentiation with BMP7 were both growing well, in vitro. Microscope observations showed that the periosteal cells were spindle-shaped, well-stacked, transparent and three-dimensional in the early stage, and cube-shaped or puncheon shaped in the mitotic phase, gradually became wide shuttle and irregular shape with a lot secretion in telophase. The positive cells were visible by the ALP kit staining and Von Kossa staining of calcium nodules at 5 d, 10 d, 15 d and 20 d in both groups.A difference of positive rate at each time point was found between BMP7 group and control group at 5 d, 10 d, 15 d, 20 d, and the difference was statistically significant (P ＜ 0.01). Conclusion It displayed well regeneration and osteogenesis ability in the periosteal cell. BMP7 has definite osteo-inductive activity, which can obviously enhance the proliferation and ossifyng differentiation of periosteal cells.

Gynaecomastia is a kind of benign and hyperplastic disease of male breast tissue. Based on the literature at home and abroad, different classification or standard of gynaecomastia has been expounded respectively from the pathogenesis, age, pathology, degree and iconography, and several common classification has been compared. Summarizing the advantages and disadvantages of these classification standards, evidence can be provided for clinical diagnosis and preoperative assessment of gynaecomastia, improvement direction can be offered for domestic classification criteria.