Objective To investigate the effects of siRNA silencing enhancer of zeste homolo 2 (EZH2) on invasion and migration of cervical cancer cells and the molecular mechanisms.Methods The small interfering RNAs (siRNA) targeting EZH2 were transiently transfected into C33A cell line by lipofectamin2000.The effects of EZH2 on cell invasion and migration were detected by wound-healing assay, Transwell assay and soft agar colony assay.The expression of MMP2 was detected by Western blot.Results Downregulation of EZH2 expression by siRNA in C33A cell line significantly inhibited cell invasion and migration in vitro.Meanwhile, siRNA-mediated depletion of EZH2 reduced the expression of MMP2.Conclusion Knocking down EZH2 expression by siRNA could surpress invasion and migration of human cervical cancer cells, which might be related to downregulating MMP2 expression.

Objective To integrate existing data and analyze the optogenetic application in the study of the cocaine addiction mechanism.Methods The key words optogenetics,addiction, cocaine and so on were selected.Relevant research information was collected from various authentic databases such as China Journal Full-text Database,Chinese Biomedical Databases,Wanfang Digital Journals,VIP Full-text Database and PubMed,SCIENCEDIRECT,SPRINGERLINK databases by literature study,assess the quality of the studies and the qualitative systematic review was conducted in this study.Results Findings from present data showed that:with great spatial and temporal specificity,the optogenetic application not only proved the important role of traditional theories of mesolimbic dopamine system,but also dissected the neural circuits and molecular mechanisms of dopaminergic and glutamatergic neurotransmission underlying cocaine addiction.Conclusion The use of optogenetic tools has given researchers the potential to dissect the neuronal circuits and molecular changes,leading us to a better understanding of the mechanisms of cocaine addiction and to the development of novel treatment.

Objective To summarize the successful experience of three cases of heart-lung transplantations performed in our institute.Methods From July 2003 to August 2012,three patients,with diagnosis of end-stage heart-lung diseases,received heart-lung transplantation in our institute.One case was diagnosed as congenital atrial septal defect,Eisenmanger syndrome,NYHA class Ⅳ; one was dilated cardiomyopathy with moderate/severe pulmonary arterial hypertension,NYHA class Ⅲ-Ⅳ,one was diagnosed as double outlet left ventricle (DOLV) with ventricle septal defect and stenosis of pulmonary artery and its left and right branches,NYHA class Ⅲ-Ⅳ.Donor hearts were preserved with UW solution,donor lungs were preserved with Euro-Collin solution in case one and with low potassium dextran containing prostaglandin E1 in the others.Extensive disinfection and strict scrutiny were implemented postoperatively.Immunosuppressive therapy included administration of zenapax or basiliximab preoperatively,methylprednisolone during the operation,and cyclosporine a/tacrolimus + prednisone + mycophenolate postoperatively.Surgical hemostasis is of great importance,as the total pleural effusion reaches 14 640 ml within 31 days postoperatively in case two.Strict postoperative disinfection and isolation were implemented,and management of the respiratory tract was intensified.Therapeutic bronchoscopy was performed frequently for sputum suction.In case two,bronchoscopy was used thirteen times within 40 days after transplantation.Broad-spectrum antibiotics and antifungal antibiotics were used for infection control.Results All three patients were discharged after recovery from operation.Case one died of obstructive bronchitis and lung failure caused by chronic rejection four years and ten months postoperatively.Case two died of sudden cerebrovascular accident 68 days after operation.Case three survives more than one year postoperatively so far and is still alive.Conclusion Proper preservation of the donor heart and lung,perfect surgical hemostasis,strict infection control,frequent application of bronchoscopy and appropriate immunosuppressive management are critical to the success of heart-lung transplantation.

Objective To estimate the application value of a standard operating procedure (SOP) in the detection of syphilitic anticardiolipin reagin. Methods Clinical laboratories from 9 local hospitals in Shanghai participated the program. Quality control samples with unknown target value were qualitatively and quantitatively examined according to the uniform SOP in these laboratories with the same reagent and facility of horizontal reaction. External quality assessment (EQA) was carried out by using seven serum samples with no, or low (1∶ 128 dilution) to high (1∶1 dilution) concentrations of target before and after the implementation of SOP. The test results were statistically analyzed and the reasons for the detecting error were assessed. Results A total of 388 tests were performed in the 9 clinical laboratories. The total accuracy rate was 93.0%, including 40.2% in the detection of samples with 1 ∶ 8 dilution of target, 49.2% in the detection of samples with 1 ∶ 16 dilution of target, and 3.6% in the detection of samples with 1 ∶ 32 dilution of target. No forward bias was observed in these tests. There was a significant difference in the accuracy rate between the two times of EQA before and after the implementation of SOP (x2 = 4.17, P < 0.05). Conclusions The improved standard procedure for nontreponemal antigen test is beneficial to the decrease of testing error, and may provide a basis for the establishment of SOP and implementation of internal quality assessment.

Objective To explore and evaluate MRI in diagnosing primary muscle non-Hodgkin lymphoma. Methods Six surgically confirmed primary muscle non-Hod#in lymphoma underwent MR examination including T_1WI, T_2WI and T_1 WI enhanced studies. The acquired images date was reviewed and analysed retrospectively in comparison with surgical and pathological results. Results The locations of 6 cases were cervical part (2), upper extremity (1), lower extremity (3), respectively. All cases involved of more than one anatomical compartment with poorly defined solid masses in 5 cases and well defined in 1 cases, 5 extended to subcutaneous fat and 3 extended along the neurovascular bundle. The mean tumor diameter was 13.9 cm, ranging from 7.3 to 22.5 cm. One was well demarcated and 5 were ill-defined. On T_1 WI, 2 were slighdy high signal intensity and 4 were slighdy low signal intensity. On T_2 WI, 2 were slightly high signal intensity, 3 were intermediate signal intensity and 1 was high signal intensity. Five were inhomogeneous and 1 was homogeneous. The intrinsic structure such as muscle fiber, tendo, spatium intermusculare were detected on 5 cases. Of the 5 dynamic contrast-enhanced cases, it showed moderate enhamcement during arterial phase, 2 were homogeneous and 3 were inhomogeneous. And it showed progressive enhancement during interstitial phase, 3 were homogeneous and 2 were inhomogeneous. Conclusions Primary muscle lymphoma always originated deep to the fascia showing subcutaneous extension and multiple compartment invasion. Typically form poorly defined solid masses with slightly high in signal intensity on MR T_2WI and middle degree dynamic delayed contrasted-enhanced in which intrinsic anatomic structure such as muscle fiber, tendo, spatium intermusculare and so on can be discerned, almost all cases involve more than one muscle compartment and some of tumor extend along the neurovascular bundle.

BACKGROUND: Growth differentiation factor-5 (GDF-5) plays an important role in the development and formation of cartilage, extremities, and joints, which is a widely used joint development marker.OBJECTIVE: To express mature peptide of human GDF-5 in E. coil by the way of genetic engineering, and to explore the inductive activity of recombinant protein in vivo.DESIGN, TIME AND SETTING: The observation experiment based on gene was performed at the Analysis and Testing Center of Southern Medical University from January to June 2006.MATERIALS: Human fetus cartilage tissue was harvested from Department of Gynaecology and Obstetrics, and the consent was obtained from the family. Ten KM mice were purchased from experimental animal center of Southern Medical University, half male and half female, weighing 18-22 g, aged 6-8 weeks.METHODS: The hGDF-5 gene encoding mature peptide was gained by RT-PCR from the total RNA which was extracted from fetus cartilage tissues, and was inserted into the pET22b(+) vector to construct recombinant prokaryotic expression plasmid pET22b(+)-GDF5, which was transformed into E. coil BL-21 to be expressed after IPTG induction. Proteins of interest were purified with sepharose chelated with nickel ions (Ni2+) and then implanted in mouse hindlimb muscle to evaluate the biological activities by routine hematoxylin-eosin staining.MAIN OUTCOME MEASURES: The expression, sequencing of target gene was observed by agarose gel electrophoresis, and the protein expression was detected by SDS-PAGE electrophoresis, meanwhile, the GDF5-inducing activity was evaluate by histological observation.RESULTS: RT-PCR product was about 350 bp in length, which was confirmed by double enzyme digestion of the recombinant plasmid, sequencing result was in agreement with the reported hGDF-5 sequence in Genbank. SDS-PAGE analysis showed a conspicuous band representing a new foreign protein with relative molecular mass of approximately 14 KD after induced expressioin. Cartilage tissues were formed in the mouse muscle where the purified proteins were implanted. CONCLUSION: The integral human GDF-5 mature peptide gene was cloned successfully from human fetus cartilage tissue and a high-yield expression was achieved in E. coli, the pudfied protein has chondrogenic activities in vivo.

Objective To Explore the imaging features relative to pathology of pelvic chondrosarcoma and to evaluate the clinical value.Methods All 12 cases patients with primary pelvic chondrosarcoma confirmed by pathological examination underwent radiography,spiral CT plain scanning,MR SE-T1WI,FSE-T2WI and SE-Tl WI enhancement scanning before operation.The imaging data was reviewed and analyzed retrospectively to compare with surgical and pathological results.Results Eleven conventional chondrosarcoma and one dedifierentiated chondrosarcoma were located in different parts of pelvis.The diameters of the tumors ranged from 4.7 to 17.0 cm with one case less than 5.0 cm,6 cases being 5.O-10.0 am and 5 cases more than 10.0 Cln.The CT valHe of 5 cases was identical or inferior to muscle with mild to moderate"ring-and-arc"mineralization and soft mass.MR imaging depict the high water content of these lesions as very high signal intensity was detected on T2 WI.Six cases showed typical"ring-and-arc"fibrous tissue which enhanced persistently.Aggressive features of deep endosteal scalloping and soft-tissue extension was also found in these cases.Conclusions Radiographic findings Can suggest the diagnosis of pelvic chondrosarcoma when there is typical"ring-and.arc"fibrous tissue,mineralization,aggressive features of deep endosteal scalloping and large soft-tissue extension.MR imaging reflect directly this pathologic structure,superior to that of CT and radiography.CT is optimal to detect the matrix mineralization,particularly when it is subtle or when the lesion is located in anatomically complex pelvic areas.

Objective To explore the imaging features of primary malignant fibrous histiocytoma(MFH)of bone and correlate them with pathological findings.Methods Thirteen cases patients with primary MFH of bone confirmed by surgical pathology underwent radiography,spiral CT plain scanning and MR SE-T1 WI,T2 WI and SE-T1 WI enhancement scanning before operation.The imaging date was reviewed and analysed retrospectively in comparison with surgical and pathological results.Results Of 13 MFH,11 were located in the end of long bone,and 2 in the diaphysis.The distance between tumors in the end of long bone and adjacent joint surface was 1 to 5 cm.All lesions showed osteolytic destruction with the maximum diameter of the tumors from 5.3 to 12.7 cm.The tumors had eccentric aggressive osteolytic destruction in 10 lesions,internal crest within the lesions in 7,inconsecutive marginal osteosclerosis in 11,little periosteal reaction in 2 and small soft tissue masses in 9,respectively.The CT value of lesions was similar to muscle.MR imaging depicted low signal intensity with aggressive features on T1 WI,iso to slight high signal intensity on T2 WI,and middle or high degree contrast enhancement on enhanced T1 WI images.Macroscopically,MFH was usually located eccentrically within the bone and produced little or no osseous expansion.The soft tissue component appeared multi-nodules and pseudo-encapsulated.Histologically,they consisted of spindle-shaped fibroblasts,which radiated outward in a spiral array from a central focus and produced a nebula or storiform appearance,and cells,which were small and oval with little visible cytoplasm.Conclusions The imaging manifestations of MFH were specific to some extent.Combined utilization of plain X-ray,CT,and MRI is helpful for the diagnosis and differential diagnosis of MFH.

Objective To investigate the effects of gene transfection with human growth/differentiation factor 5(GDF5)on the growth and difierentiation of bone nlarrow stromal stem cells (BMSCs).Methods GDF5 gene was trans fected into BMSCs by liposome method. Then cell proliferation and cycles were examined by MTT and flow cytometry respectively. Cell morphology was observed under light microscope and electron microscope (EM).GDF5 and Collagen Ⅱ were detected at the level of mRNA and protein by RT-PCR and immunocytochemistry. Alkaline phosphate activity was examined by lead citrate method. Osteocalcin mRNA expression was determined bv RT-PCR. ResulIs GDF5 gene was transfected into BMSCs successfully and the transfected cells still maintained their natural growth and proliferation features. Stable expression of GDF5 gene in BMSCs was obtained. The trans fected ceils had basically the same proliferation ability and cell cycles as the untransfccted ones. After transfection comparatively more polygonal cells could be seen in light microscope, showing irregular arrangement mode. Plenty organells were observed and cell nucleus showed irregular shape under EM. The expressions of Collagen Ⅱ mRNA and protein were positive. But osteocalcin mRNA expression was negative. Conclusion Since BMSCs can be induced by GDF5 to differentiate into chondrogenic cells. GDF5 gene-modified BMSCs may be used as candidate seed cells of cartilage tissue engineering.

Angiography ; Female ; Hepatic Artery ; abnormalities ; diagnostic imaging ; Humans ; Middle Aged ; Tomography, X-Ray Computed ; Ultrasonography