Objective To examine the analgesic effect of intrathecal (i.t.) adenovirus containing human beta-endorphin (?-EP) gene in a rat model of neuropathic pain produced by chronic constrictive injury (CCI) . Methods Thirty-six male SD rats weighing 210-260 g were randomly divided into 3 groups: (1) blank control group (n = 5); (2) sham-operated group (n = 5) ; (3) neuropathic pain group (n = 26) . The neuropathic pain group was further divided into 3 subgroups: Ad-NEP subgrouop (n = 9); Ad-GFP (the recombinant adenovirus containing green fluorescent protein) subgroup (n = 9) and normal saline (NS) subgroup (n = 8). The animals were anesthetized with intraperitoneal ketamine 100 mg?kg-1 and atropine 50 mg?kg-1. Neuropathic pain was produced by ligation of right sciatic nerve according to the technique described by Bennet and Xie. In sham-operated group the sciatic nerve was exposed but not ligated. In blank control group no operation was performed. Seven days after the surgery a PE-10 catheter was placed in the subarachnoid space at L5,6 according to the method of Milligan et al. Seven days after catheter placement 1?108 pfu Ad-NEP, Ad-GFP and 0.9% saline were injected i.t. via the catheter respectively. The paw-withdrawal latency to radiant heat was measured before surgery (T0,baseline) , the day of i.t. injection (T1) and 1 day (T2) , 1 w (T3), 2 w (T4), 3 w (T5), 4 w (T6) 5 w (T7) after i.t. injection. At one week after i.t. administration one animal in Ad-NEP subgroup and Ad-GFP subgroup was killed and the lumbar segment (L3-6) of the spinal cord was removed for immuno-histochemical examination. Naloxone 1 mg?kg-1 was given intraperitoneally in Ad-NEP subgroup (n = 8) and Ad-GFP subgroup (n = 8) at 10 days after i.t. injection. Pain threshold to thermal stimulation of the right paw was measured before (t0) and from 10-90 min after intraperitoneal naloxone injection at an interval of 10 min (t1-9). CSF samples were obtained at T1-7 for determination of CSF concentration of ?-EP using radio-immunological assay. Results The right paw-withdrawal latencies (PWLs) were significantly lower in Ad-GFP subgroup and NS subgroup than in blank control and sham-operated groups (P

Objective To observe the morphologic changes of of vascular buds in vertebral cartilage endplate in age-specific rabbits and also to investigate the correlation between the changes of vascular buds and interverbral disc degeneration. Methods There were 15 New Zealand white rabbits in our study,which include three groups, 2-week-old rabbits, 1-year-old rabbits and 3-year-old rabbits, and each groups had five rabbits. The X-ray radiograph, histology and scanning electron microscope were used to observe the changes of vertebral cartilage endplate. According to Miyamoto standard, the interverbral disc was graded 1-5, and scored 1-5 respectively. Results The changes of micro-vascular structure of vertebral cartilage endplate were observed during aging. Under the scanning electron microscope, the vascular structure degenerated gradually, and disappeared in the end. The blood vessels in the central region of the vertebral cartilage endplate reduced more obviously than those in periphery region. The severe degeneration was found in vertebral endplate, compared with intervertebral disc. The changes of vascular buds in rabbits vertebral cartilage endplate had positive correlation with the vertebral endplate calcification and the interbertebral disc degeneration. Conclusion Changes of vascular buds in vertebral endplate may accelerate intervertebral disc degeneration.

Objective:To clarify whether the signal peptide of human nerve growth factor can mediate secretory expression of beta-endorphin and whether there is difference between the efficiency of signal peptides from human and mouse nerve growth factor.Methods: Two kinds of eukaryotic vectors containing human or mouse signal sequence-mediated secretory expression of beta-endorphin were constructed.The culture supernatant and cells were collected 48 h after NIH3T3 cells were transfected by the two kinds of vectors,and the cover slips with single-layer cells was prepared.The concentration of beta-endorphin in the culture was determined by radio-immunoassay.The total RNA was extracted from cells and mRNA from fusion genes was assayed by RT-PCR.Cells on cover slips were subjected to immunofluorescence staining.Results: RT-PCR showed that the fusion genes were expressed in NIH3T3 cells;the expression of beta-endorphin was mainly in the cytoplasm of NIH3T3 cells.The concentrations of beta-endorphin in the supernatants 48 h after transfection with pcDNA3.1-hEP and pcDNA3.1-mEP were(280.33?24.16) pg/ml and(191.04?7.96) pg/ml(P

Objective To investigate the clinical significance.of thrombocytopenia in systemic lupus erythematosus (SLE).Methods One hundred and two SLE patients with thrombocytopenia who were admitted to Peking University People's Hospital were involved in the study.SLE patients without thrombocytopenia were controls.Clinical and laboratory characteristics were analyzed.T-test and Chi-square test were used for inter-group comparison.Results Patients with thrombocytopenia had more organ damage than those without,although the disease activities (SLEDAI) were not different between these two groups.Bone marrow characteristics were analyzed and 16 patients were amegakaryocytic.However,there were no differences observed between patients with amegakaryocytosis and normal megakaryocytes in organ damage,disease activity and response to therapy.Conclusion Lupus patients with thrombocytopenia usually have more organ damage.About 32％ of those patients are amegakaryocytosis.

Objective To describe an ideal technique to repair full-thickness lower lid marginal defects in a one-stage procedure. Methods The buccal rotation flap and the nasal septal chondromucosal flap were used in one-stage operation to repair full-thickness lower eyelid defect. Results Eleven patients including 6 male patients and 5 female patients underwent lower eyelid reconstruction since January 2000.The age ranged from 29 to 64 years with average 43. Seven patients with full-thickness lower eyelid defect were caused by basal cell carcinoma excision, while four patients were caused by trauma. All the fullthickness lower eyelid defects were reconstructed by using the buccal rotation flap and the nasal septal chondromucosal flap in one-stage operation. All the flaps survived completely after operation. There were no other complications excepting that two patients developed to mild lower eyelid retration after six months. Conclusion The technique consisting of the buccal rotation flap and the nasal septal chondromucosal flap is a simple and useful alternative procedure to close full-thickness defects in the lower eyelid.

Objective To study the role of calcium homeostatic and kinetics in the epileptogenesis activity. Methods Hippocampal neurons were acutely isolated from controls and status epilepticus (SE) models induced by lithium-pilocarpine at different time point. The [Ca2+]i levels were detected by laser scanning confocal microscope. And the ability to restore resting [Ca2+]i levels after a brief exposure to 5 μmol/L glutamate in control and epileptic neurons were evaluated. Results The [Ca2+]i level of acute separated hippocampal neurons in the control rats was (95.4±22. 1) nmol/L After injection of lithium pilocarpine, the [Ca2+]i level in hippecampal neurons increased dramatically to (867.6±35.2) nmol/L, and decreased to (292.8 ± 18.3) nmol/L on the 7th day, lasting for about 30 days ((220. 8± 17.6) nmol/L), it is higher than that in the control group (t = 12. 55, P < 0.01). The distribution of neuronal [Ca22+]i showed that 92% of control neurons were in the normal range of [Ca2+]i level (25-150 nmol/L) ; After 6 hours, however [Ca2+]i levels of all SE neurons increased, and 85% of which were higher than 500 nmol/L; After 7, 14 and 30 days, there were 75%, 60% and 52% of SE neurons still manifested an elevated [Ca22+]i level, but less than 500 nmol/L. After the exposure to 5 μmol/L glutamate treatment for 2 minutes, [Ca2+]i of the control neurons restored to baseline values in (9. 5±3.4) minutes, whereas the SE rats of acute, latent and chronic phases did not (t = 5.08, 4. 56, 4. 21, all P < 0. 01). Conclusion Lithium-pilocarpine induced epilepsy causes a long-term alteration of calcium homeostatic mechanisms of hippocampus neurons, which may play an important role in the development and maintenance of spontaneous recurrent seizures.

Objective ① To Screen for the miRNAs differently expressed in the peripheral blood mono-nuclear cells (PBMCs) of rheumatoid arthritis (RA) by microarray experiments.② To further evaluate the expression of miR-155 in PBMCs of RA.③ To determine the relevance between the expression of miR-155 and clinical as well as laboratory features.④ To test whether inflammatory mediators can induce miR-155 expression in PBMCs of RA.Methods ① Total RNA was isolated from peripheral blood mononuclear cells obtained from 5 patients of RA and 5 normal controls.Expression profiling of miRNAs was performed in a microarray analysis.② MiR-155 was identified for further study by stem-loop real-time RT-PCR based on SYBR-Green.PBMC from 26 patients of RA and 23 normal controls were collected.③ Association between miR-155 and the clinical and laboratory features of RA was evaluated.④Induction of miR-155 following stimulation with TNF-α, IFN-γ and LPS of cultures of RA PBMCs was examined by real-time RT-PCR.Statistical analysis was done with student's t test, paired t test, and ANOVA, Spearman correlation.Results ① Expression profiling of miRNAs revealed significant differential expression of 14 miRNAs, of which signal intensity changed over two times.MiR-155 was up-regulated in PBMCs of RA than in normal controls (t=9.218, P=0.001).② The expression level of miR-155 had a positive correlation with serum CRP level (r=0.57, P=0.002).③ Expression of miR-155 was markedly up-regulated in PBMCs of RA after stimulated with TNF-α,IFN-γ and LPS, especially with TNF-α.Conclusions The expression of miR-155 is induced by stimulating with TNF-α, IFN-γ and LPS.MiR-155 may be a regulator in RA pathogenesis.Further studies are required to elucidate the function of miR-155.

Objective To prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering using porcine fibrin and decellularized canine carotid artery.MethodsPorcine fibrin was sprayed coating on the external surface of decellularized canine carotid artery to construct completely biological hybrid scaffold for small-diameter vascular tissue engineering. The completely biological hybrid scaffold was evaluated with Hematoxylin and Eosin (H&E) staining, scanning electron microscopy and biomechanics test.ResultsHistology examination revealed that the porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery. Scanning electron microscopy examination confirmed that the external surface of completely biological hybrid scaffold was smooth and uniformly. Compared with fresh canine carotid artery and decellularized artery, the biological hybrid scaffold had similar burst and breaking strength. Furthermore, compared with decellularized artery, the biological hybrid scaffold had higher compliance.ConclusionThe porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery to prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering. The biological hybrid scaffold had appropriate biomechanical properties and had potential to serve as scaffolds for small-diameter vascular tissue engineering.

Objective To prepare completely biological tissue-engineered small-diameter blood vessel based on a biological hybrid scaffold.MethodsEndothelial cells and smooth muscle cells were isolated from the porcine aorta and expanded in vitro. Mixture of smooth muscle cells and porcine fibrin was prayed coating on the decellularized canine carotid artery. Then, the inner surface of the decellularized artery was seeded with the endothelial cells to construction of completely biological tissue-engineered small-diameter blood vessel. The tissue-engineered blood vessel was evaluated with Hematoxylin and Eosin (H&E) staining and scanning electron microscopy.ResultsHistology examination revealed that the completely biological tissue-engineered small-diameter had intact media and intima. Scanning electron microscopy examination confirmed that the inner surface of tissue-engineered blood vessel was covered with intact monolayer endothelial cells and the external surface was covered with multilayer smooth muscle cells.ConclusionThe completely biological tissue-engineered small-diameter with intact media and intima was prepared using mixture of blood vessel cells and porcine fibrin on the decellularized canine carotid artery.