Objective To construct the nuclear receptor-related factor 1 ( Nurr1) specific short-hairpin RNA (shRNA) expressing vectors and study their effects on the expression of endogenous genes in MN9D dopaminergic cell lines. Methods Two RNAi recombinant plasmids (named pSC-N1 and pSC-N2) targeting at Nurrl gene were constructed. The purified plasmids were identified by DNA sequencing. After being transfected into MN9D dopaminergic cell lines with Lipofectamine?2000, the silencing effects of Nurrl shRNA expressing vectors on the expression of Nurrl mRNA and protein in MN9D cells were detected using real time fluorescence quantitative PCR and Western blot Results DNA sequencing confirmed that the Nurrl shRNA expressing vectors were constructed successfully. Nurrl mRNA expression in MN9D cells was specifically suppressed after the transfection of pSC-Nl and pSC-N2 by 62.3% and 45.6% respectively. Moreover, the expression of Nurrl protein was significantly suppressed by 57.4% and 72. 0% respectively. The control vectors had no silencing effect Conclusions The vectors pSC-Nl and pSC-N2 have been successfully constructed and can specifically suppress the expression of Nurrl mRNA and protein. Nurrl specific shRNA expressing vector may provide a novel applicable strategy for the study of the function of the genes associated with Parkinson disease and the development of dopaminergic neuron.

Objective To investigate the clinical features of acquired immunodeficiency syndrome (AIDS) with neurological complications. Methods Five with neurological complications out of 31 patients with confirmed AIDS in Shanghai area from 1992 to 2001 were analyzed. Results Of the five cases, there were one complicated with vacuolar myelopathy, one vacuolar myelopathy accompanied with dementia, one cerebral tuberculoma accompanied with dementia, one cerebral infarction accompanied with symptomatic epilepsia and trigeminal neuralgia, one polyneuropathy. Two of the vacuolar myelopathy are those showing first occurring manifestations. Conclusion AIDS patients are found at high risk of neurological complications, of which vacuolar myelopathy and AIDS related dementia are more common. If young patients who suddenly suffered from dementia, especially in patients accompanied with opportunistic infections, AIDS should be considered. In these cases serum HIV antibodies should be checked.

Objective To explore the effect of glucose regulated protein (GRP)78 on the neuronal apoptosis after cerebral ischemia reperfusion injury in rats. Methods Middle cerebral artery ischemia reperfusion rat's models were used with the modified filament method. The expression of GRP78 in the ischemic penumbra tissue was detected by RT-PCR, Western blot and immunohistochemistry at different time points. Primary cultured rat's neurons were exposed to hypoxia and subsequently reoxygenation. Western blot was used to investigate the expression of GRP78. The changes of the neuronal apoptosis after overexpression of GRP78 induced by 2-deoxyglucose were detected. Results The expression of GRP78 in the ischemic penumbra tissue in model group was significantly increased (mRNA : 0.7367±0.0651, F= 477.160, P < 0.01 ; Protein : 0. 8129±0. 0748, F=39.857, P < 0.01). The neuronal survival status was increased after overpression of GRP78 (increased by 39.22% ± 0. 44%, t=46.374, P < 0.01) while the neuronal apoptosis was decreased (decreased by 16.60±1.02, t=7.530, P <0.01). Conclusion Focal cerebral ischemia reperfusion can induce endoplasmic reticulum stress which plays a role in the neuronal apoptosis. The increased expression of GRP78 may protect the ischemic tissue from neuronal apoptosis.

Objective To evaluate the value of neutrophil gelatinase - associated lipocalin (NGAL) for diagnosis of acute kidney injury (AKI) after cardiopulmonary bypass established. Method 120 children undergoing cardiopulmonary bypass were enrolled in this prospectively study. Serial urine samples were analyzed by ELISA for NGAL expression. The primary outcome measure was AKI, which was defined as a 50% or greater increase in serum creatinine from baseline. Result 21 children ( 17.5% ) developed AKI. Urine concentrations of NGAL increased after cardiopulmonary bypass had been established for 2 hours. The concentration of NGAL in urine after cardiopulmonary bypass had been established for 2 hours was used to diagnose AKI. The area under the receiver-operating characteristic curve was 0. 96, the sensitivity was 92%, and the specificity was 91% for the cutoff value of 100ng/mg Cr. Conclusion Concentrations of NGAL in urine might be used as a sensitive and specific biomarker for early prediction of AKI after cardiopulmonary bypass had been established.

Male germ-line stem cells (mGSCs) have the capability of self-renewal and latent capability of differentiation. mGSCs is the unique diploid immortal cell which can transfer genetic information to filial generation. The combination of transgenic technology and mGSCs heterotransplanting will supply new opportunities and paths to cloning animal, transgenic animal and gene therapy of some human hereditary disease. We studied the isolation and cultivation of mGSCs that were isolated and purified from 5-6 month old bovine fetal testis, new born bovine testis by adopting mixed enzymes digestion and different attaching velocities methods. The results showed that Sertoli cells were indispensable to mGSC's proliferation and differentiation in vitro. The Sertoli cells in logarithmic phase have a significant effect on mGSC's attaching, proliferation and differentiation. Co-culture with Sertoli cells, mGSCs differentiated to long sperm after 16 days. A preliminary system for mGSC's inducing differentiation was established.
Animals ; Cattle ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Male ; Sertoli Cells ; cytology ; Spermatozoa ; cytology ; Stem Cells ; cytology ; Testis ; cytology

Objective To investigate the correlation between IL‐18 and deep venous thrombosis disease and its clinical significa‐tion .Methods To detect the expression of IL‐18 by ELISA ,we collected the blood samples of DVT patients as the experimental group(n=40) compared to the control group(n=40) and normal group(n=20) .IL‐18 over expression/interference vectors were constructed and transfected human vein endothelial cells ,analyzed by microarray and KEGG Pathway as biology information tech‐nology .Then discuss the association between IL‐18 and DVT .Results Results of ELISA showed that compared with control group and normal group ,the expression of IL‐18 gene in DVT patient were up‐regulated(F=11 .248 ,P<0 .01) .Compared with normal group ,the IL‐18 expression in control group have not been significantly up‐regulated(P>0 .05) .Immunofluorescence detected IL‐18 gene expression in cytoplasm of human umbilical vein endothelial cells (HUVECs) .According to the microarray analysis we found in the IL18‐pCDH‐GFP transfected cells 17 signaling pathways were down‐expressed while 16 signaling pathways were up‐expressed .Compared with normal group cells ,in the IL18‐LMP‐shRNAmir1 transfected cells 23 signaling pathways were down‐ex‐pressed and 9 signaling pathways were up‐expressed .Conclusion Based on the above experimental data ,it is very clear that IL‐18 influenced HUVECs and plays an important role in DVT ,it is possible to predict the diagnosis of DVT and act as candidate molecu‐lar markers .

Domesticated ungulates embryonic stem (ES) cells have great significances in biology and wide application prospects. This review compared the key signaling pathways related with pluripotency between mouse and human ES cells, and the difference of transcription factors in mouse, human and domesticated ungulates ES cells were elaborated. Finally the pluripotency candidate signaling network and transcription factors related in the derivation of domesticated ungulates ES cell were discussed combined with practical experience of ovine embryonic stem cell derivation in our laboratory.
Animals ; Animals, Domestic ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; metabolism ; Humans ; Leukemia Inhibitory Factor ; metabolism ; Mice ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Pluripotent Stem Cells ; cytology ; metabolism ; Sheep ; Signal Transduction ; Transcription Factors ; metabolism ; Wnt Proteins ; metabolism

Objective:To develop a role model education intervention model for male nursing students, and to explore the effect of role model education on male nursing students′ professional identity and positive psychological capital.Methods:A total of 60 male nursing freshmen were recruited and randomly assigned to the control group (n=30) and the experimental group (n=30) according to a 1:1 ratio. The control group only accepted the school′s routine teaching tasks and other campus activities. The experimental group received 6-week model education intervention on the basis of the control group once a week. Nursing students′ Professional Identity Questionnaire and Positive Psychological Capital Questionnaire were used for assessment at the time of enrollment and after intervention.Results:Male nursing students' professional identity (50.25±11.19) and positive psychological capital (114.68±29.08) were generally not high. After intervention, the scores of occupational self-concept, retention benefits and resignation risks, social persuasiveness and total score in the experimental group were 22.64±5.19,14.29±2.69,8.00±1.89,64.18±12.15, higher than those of the control group (20.07±3.16, 12.93±1.76, 6.86±1.24, 57.68±6.66), and the difference was statistically significant ( t value was 2.23-2.68, P<0.05). Conclusions:Role model education intervention has a good effect in promoting professional identity and positive psychological capital of male nursing students.

Muscle is one of the most important tissues of human body, which is distributed around various organs and bones. Skeletal muscle plays an important role in human activities and its functional changes are closely related to its own morphological structure. The study of the relationship between musculoskeletal structure and function can help us to understand the physiology basics of force and to guide clinical practices. Ultrasonography has been widely used in the research of muscle properties since it is real-time, fast, nonradiative and inexpensive. In recent years, there emerges various researches on image processing method for musculoskeletal ultrasonography, especially for dynamic ultrasonography. This paper presents a brief overview of the existing methods and key steps of ultrasound image processing of musculoskeletal.
Humans ; Image Processing, Computer-Assisted ; Muscle, Skeletal ; diagnostic imaging ; Ultrasonography

Objective: To explore the effects of gentiana scabra bage on the expression of hepatic collagen proteins in Paragonimus skrjabini rats with liver fibrosis. Methods: Immunohistochemical technique was used to observe the changes of content of hepatic type I, III collagen proteins in Paragonimus skrjabini rats with liver fibrosis before and after the gentiana scabra bage treatmeat. Results: Comparing with the model group, changes of hepatic type I and type III collagen proteins in gentiana scabra bage treated group were significantly weakened. Conclusions: Gentiana scabra bage treatment can reduce the content of hepatic type III and type I collagen protein significantly in Paragonimus skrjabini rats with liver fibrosis, thereby, playing the role against hepatic fibrosis.