Objective:To investigate the effect of NF-κB p65siRNA of IL-17 inducing the expression of MCP-1 in the primary cultured cardiac myocytes.Methods:The cardiac myocytes were isolated from neonatal mice by different adhesion method.NF-κB P65 siRNA was transfected into cardiac myocytes and the rates of transcription and translation of MCP-1 were detected by RT-PCR and enzyme-linked immunosorbent assay( ELISA) ,the rates of transcription and translation of P-P65 and P65 were detected by RT-PCR and Western blot.Results:Compared with negative siRNA group,the expression of the MCP-1 at mRNA and protein levels were increased in negative siRNA +IL-17 group(P<0.05).Compared with negative siRNA group,the expression of the MCP-1 at mRNA and protein levels were decreased in NF-κB P65 siRNA group( P<0.05).Compared with negative siRNA+IL-17 group,the expression of the MCP-1 at mRNA and protein levels were decreased in NF-κB P65 siRNA+IL-17 group(P<0.05).The expression of the NF-κB P65 at mRNA and protein levels in cardiac myocytes were specifically and extensively suppressed by NF-κB P65 siRNA(P<0.05).After stimulated by IL-17,the amount of P-P65 in the cardiac myocytes was significantly increased in time-dependent manner compared with that of black group( P<0.05) , but the levels of P65 changed little ( P>0.05 ) .Conclusion: IL-17 stimulates MCP-1 expression in cardiac myocytes via NF-κB activation,and NF-κB P65 siRNA can effectively inhibit the upregulation of IL-17 on MCP-1.

Adolescent ; Humans ; Magnetic Resonance Angiography ; Magnetic Resonance Imaging ; Male ; Sinus Thrombosis, Intracranial ; diagnosis

Animals ; Dose-Response Relationship, Drug ; Female ; Humans ; Metals, Heavy ; analysis ; pharmacology ; Placenta ; chemistry ; drug effects ; physiology ; Pregnancy ; Rats

Objective To establish the model of amyotrophic lateral sclerosis (ALS) with selective motor neuron disorder by organotypic spinal cord cultures, and analyze the role of astrocyte in the pathagenisis of ALS.Methods Organotypic spinal cord cultures were prepared using lumbar spinal cord slices from 8-day-old SD rat pups. The threohydroxyaspartate (THA) was applied into culture medium to establish ALS organotypic spinal cord cultures model. Motor neurons survival was evaluated by monoclonal SMI-32 immunohistochemical staining and glial fibrillary acidic protein staining to show astrocyte survival.Results Compared with the control group, there was significantly astrogliosis in the anterior horn and surrounding white matter in THA 100 μmol/L group, and the level of gliosis was increased followed the elongation of THA interference time. With the increasing of the number of astrocyte, the morphology of astrocyte was changed.Conclusion There is significantly astrogliosis in the anterior horn and the time of astrogliosis is markedly earlier than the time of motor neuron loss in the ALS model intervented with THA.

Objective To investigate the role of the Wnt/LRPS/3-catenin signaling pathway in the pathogenesis of postmenopausal osteoporosis.Methods Fifty female Wistar rats aged 6-month-old,were randomly divided into control group ( NS,n =24 ) and ovariectomized group ( NOVX,n =26),NOVX underwent bilateral ovariectomy.At 0,4 and 8 weeks,all of rats were measured blood estrogen ( E2 ) and bone mineral density (BMD),4 and 8 weeks,low density lipoprotein receptor-related protein 5 (LRP5),3-catenin and Runx2 mRNA in bone were measured respectively by reverse transcription (RT)-PCR.Results In 4 and 8 weeks,compared with NS which had (117±29) and (114+15) pmol/L in E2 level,(0.098 ± 0.016 ) and (0.095 ± 0.028 ) g/cm2 in BMD,NOVX had significantly decreased to ( 92±15 ) and(95±22) pmol/L in E2 level ( P＜0.05 ),( 0.076 ± 0.016) and ( 0.052 ± 0.013 ) g/cm2 in BMD values ( P ＜ 0.01 ).And bone tissue LRP5,β-catenin and Runx2 mRNA expression was 1.02 ± 0.06,1.04 ±0.05,1.07 ±0.21 in NS,NOVX was significantly reduced to 0.97 ± 0.04,0.58 ± 0.05,0.86 ±0.03 (P＜0.05).Conclusion Wnt/LRP5/β-catenin signaling pathway may be important in the pathogenesis of postmenopausal osteoporosis.

Objective To study further the etiology of cerebral palsy(CP),and the clinical effects of early diagnoses and intervening treatments.Methods The causes,clinical typing and CT/MRI features of CP in 240 cases were analyzed.The patients with CP were treated with the drugs of trophic nerve,the channel pilot frequency,the laser transmission and the drugs penetrating skin by point combined with functional exercise attaining ect.Results Among 240 cases in the study,birth asphyxia was 44.6％ .The total effective rate was 87.1％ . Conclusion Most cases may be prevented by health care in perinatal period.The younger the age,the more satisfactory is the therapeutic effectiveness for treating CP.

AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Objective: To observe the efficacy of mirtazapine in the treatment of depression after cerebral infarction and its effect on rehabilitation of neurological functions. Methods:117 patients with acute cerebral infarction comorbid with major depression were randomly allocated to two groups treated with mirtazapine (57 cases) or not (60 cases). Hamiltion Depression Rating Scale (HAMD), Zung's Self-rating Depression Scale(SDS), modified Edingburgh-Scandinavian Stroke Scale (SSS) and Activity of Daily Living(ADL) were measured at baseline, 2 weeks, 4 weeks, 8 weeks and 6 months after randomization.Results:At the end of 6 months trial, the effective rate for depression of mirtazapine group was 100%, including 41 with relief (41/57, 71.9%); while that of control group was 13.4% (3/60), with only 4 with relief (6.7%). For neurological function, 78.9% (45) patients in mirtazapine group had significant improvement, that number in control group was 31 (51.7%). From the third week, patients in mirtazapine group had better ADL results than baseline (31.2?11.2/39.2?15.8), at the end of 6 months, their activity of daily life was much better than that of control (15.7?5.4/21.8?9.7, t=4.17,P

Objective To construct a recombinant lentiviral vector for HCV core-Ant and then study its effect on the transdifferentiation of hepatic stem cells. Methods The HCV core-Ant was obtained by PCR of two primers template with each other and T-vector cloning method,and then subcloned to pLenti6/V5-D-TOPO. The restriction endonuclease and T4 DNA ligase were used to construct the vector. pLenti6/V5-D-HCV core-Ant and the ViraPowerTM PackagingMix (containing three packaging plasmids pLP1,pLP2 and pLP/VSVG) were cotransfected into 293ET cells to produce replication in competent lentivirus after transfection. The viral supernatant on 293T cells was collected. The expression of the lentiviral vector containing HCV core-Ant in Hela cells was measured by immunohistochemistry. Then we constructed the lentiviral vector containing green flurosecent protein by the similar method,and the titers were determined. Results HCV core-Ant was identified and analyzed by restriction enzyme digestion and sequencing,respectively. The expression of the recombinant lentiviral vector plasmid containing HCV core-Ant in Hela cells was confirmed by immunohistochemistry. The 3T3 cells transfected the lentiviral vector containing green flurosecent protein were found to show strong expression of GFP,which confirmed that the four-plasmid system of the lentiviral vector was successfully constructed. Conclusion The recombinant lentiviral vector expressing HCV core-Ant was successfully constructed by molecular cloning and recombination techniques in vitro,which will be beneficial to guiding further study on gene therapy of cancer.

Objective: To investigate the effects of the exogenous testosterone propionate on apoptosis of rat germ cells and the mechanisms thereof. Methods: Thirty 35-day-old male SD rats were randomly divided into experimental group and the control group. The rats in experimental group were injected (i.m.) testosterone propionate and the control group with an equal volume of saline. By using terminal deoxynueleotidy transferase nediated dUTP nick-end labeling (TUNEL), flow cytometry (FCM), radioimmunoassay (RIA) and electron microscopy, the quantity and quality of apoptosis of germ cells were evaluated. Results:(1) Compared with the control, the apoptotic number of rat germ cells was increased in the experimental group, especially the primary spermatocyte. The apoptotie rate was 11.3% detected by FCM in experimental group,while 3.6% in the control group (P ＜ 0.01). (2) The percentages of 1C were 21.8% in experimental group and 33.8% in control group (P ＜ 0.01).The percentages of 2C were 52.6% in experimental group and 37.1% in control group (P ＜ 0.01). (3) The serum levels of testosterone were (3 486.8±333.3) ng/L in experimental group and (846.9±167.5) ng/L in control group (P ＜ 0.01). The serum levels of follicle-stimulating hormone (FSH) were (2.5±0.8) IU/L in experimental group and (5.2±1.7) IU/L in control group (P ＜0.01). Conclusion: The exogenous testosterone propionate might induce apoptosis of germ cells by retroinhibition of the hypothalamie-pituitary-gonadal axis, thus having contraceptive effects.