Objective To investigate the effects of ropivacaine on the whole cell sodium currents in rat sympathetic ganglion neurons in order to determine whether sympathetic ganglion is involved in the ropivacaine cardiotoxity. Methods The sympathetic neurons were enzymatically isolated from the superior cervical ganglion. The effects of ropivacaine on the whole cell sodium channel currents were assessed using patch clamp technique. Results Ropivacaine blocked the whole cell sodium channel currents dose dependently. When the holding potential (Vh) was -80mV and Vt Omv , 0.01?mol/L ropivacaine reduced peak currents by 30.02% with a mean IC50 of 2.68?mol/L. The blockade was also potential dependent. When Vh was -60mv, the mean IC50 was 1.55?mol/l. 1.0?mol/L ropivacaine reduced the peak value of I V curve by 30.66% but did not affect the shape of I V curve and caused 2.56mv shift of voltage dependence activation curve to depolarized potentials and 3.56mv shift of steady state inactivation curve to more hyperdepolarized potentials.The conditioning pulse potential at which half maximal channels were activated (V1/2), changed from -52.99mv to -56.44mv and the test potential at which half maximal channels were activated (V1/2), changed from -25.2mv to -22.64mv. Conclusions Subconvulsive concentration of ropivacaine significantly inhibits sympathetic ganglion neurons in a dose dependent and potential dependent way through the inactivation of sodium channel,indicating that sympathetic ganglion neurons may contribute to the cardiotoxity of ropivacaine.

Objective To serially examine virus carrying condition of clinical specimens collected from SARS patients, in order to evaluate the dynamic changes of virus in the bady and its clinical significance. Methods A total of 494 samples, including plasma, urine, feces, sputum and throat swab, obtained from 84 cases of clinically diagnesed SARS patients at different time-points were examined for the study. Nested RT-PCR was employed for the detection of SARS-CoV using 2 pairs of primers targeting P and N region of the viral genome. The amplified viral genomic fragments were confirmed by DNA sequence analysis. Results Specific bands for SARS-CoV were amplified from various specimens of some SARS patients. Highest positive rate was found in sputum compared with the others. However, simultaneous examination of more than two specimens increased detectable rate of the virus. Moreover, dynamic examination revealed that the highest positive rate occurred in the 2nd week after the onset of the disease and next to the highest during the 3rd to 4th week. However, dynamic detectable rates for different specimens were not identical. Conclusion Nested RT-PCR is a practicable method for the detection of SARS-CoV and helpful for the early diagnosis or confirmation of the disease. Dynamic examinations for SARS-CoV by combined employment of the N and P primers and by involving multiple specimens may significantly increase the positive detectable rate, therefore would be helpful in diagnosis and eratuation of therapeutic effect of SARS.

Objective To observe dynamic responsive regularity of specific IgM and IgG antibodies in SARS patients. Methods 145 specimens of plasma from 25 cases of clinically diagnosed SARS patients were examined in the study. ELISA was employed to detect IgM and IgG antibodies against SARS coronaviral antigens. Nested RT-PCR was used to qualitatively determine SARS coronavirus (SARS-CoV). Results 49.0% (71/145) and 54.5% (79/145) of the samples were positive for IgM and IgG antibodies, respectively. Both antibodies were found to be detected in 84.0% of the patients. The antibodies were found to be detectable from the 2nd to 4th week after the onset of disease in most patients, and there was a fendeney of sising in positive rate until the 5th week after the onset. Thereafter, the detectable rate of IgM antibody began to decline, while that of IgG antibody remained to rise. Positive rate for serum SARS-CoV was 15.8% (18/114) for all samples or 40.0% (10/25) of patients. Most virus-positive samples were those which were collected within 4 weeks after disease onset. Conclusions Anti-SARS-CoV IgM/IgG antibody and detection of virus in plasma could serve as practical diagnostic indicators for SARS. In most cases, when the serum was pasitive for antibody the serum virus positive rate would soon declined. However, concomitant existence of antibody and viral sequence in plasma was observed in a few patients.

BACKGROUND: Peripheral nerve injuries are hoped to promote the regeneration of nerve repair by elevating brain-derived neurotrophic factor (BDNF) concentration in local injury regions. OBJECTIVE: To observe the recanalization of nerve fiber and motor function in sciatic nerve injury rats following BDNF gene-modified bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: The randomized, controlled animal study was performed at the Fujian Institute of Neurology from May 2007 to May 2008. MATERIALS: Femur and tibia of F344 male rats aged 2 months were sterilely harvested to prepare BMSCs. BDNF gene-modified BMSCs were prepared using constructed chronic viral vector PNL-BDNF-IRES2-EGFP. METHODS: The right sciatic nerve injury models were made using 60 adult Sprague Dawley rats. All models were randomly assigned into phosphate buffer saline (PBS) group, BMSC group and BDNF gene-modified BMSC group. 2 ?L BPS, 2 ?L BMSC solution and 2 ?L BDNF modified BMSC solution were separately transfered into injury site of each group. MAIN OUTCOME MEASURES: The retrograde horseradish peroxidase tracing was practiced to observe neural cell number in the Lumbar 4 and 5 spinal cord anterior horn at 2 and 4 weeks after microinjection. Sciatic nerve function index was used to observe rat motor function of injured limbs. Fluorescence excitation and immunohistochemistry were employed to detect BMSC survival and BDNF expression. RESULTS: Cell number was more, and nerve function recovery was better in the Lumbar 4 and 5 spinal cord anterior horn in the BDNF gene-modified BMSC group than in the PBS and BMSC groups. BMSC survival was found in the injured sites in the BMSC and BDNF gene-modified BMSC groups. BDNF expression was significantly more in the BDNF gene-modified BMSC group than in the BMSC group. CONCLUSION: BDNF gene-modified BMSCs have promotion effects on the recanalization and functional recovery of nerve fiber following peripheral nerve injury.

Aim The effects of diazepam on the whole cell sodium currents in rat sympathetic ganglion neurons were studied to investigate the mechanisms by diazepam mediates hypotension. Methods Whole cell patch clamp recordings were performed on enzymatically isolated rat superior cervical sympathetic ganglion neurons. Results Diazepam dose dependently blocked the whole cell sodium currents. Under a V t of 0 mV and a V h of 80 mV 0.3 ?mol?L -1 diazepam reduced sodium peak currents by 14.76 %(P

AIM: To study the interactions of propofol and midazolam on the whole cell sodium channel currents in rat sympathetic ganglion neurons. METHODS: Whole cell patch clamp recordings were made from enzymatically isolated rat (7- 10 d ) superior cervical sympathetic ganglion neurons. Isobolographic analysis was applied to evaluate the potency of combinations of propofol and midazolam on Na + channel currents. RESULTS: Under V h= 80 mV and V t= 0 mV . Propofol and midazolam dose dependently blocked Na + currents with a mean drug concentration required to produce 50% current inhibition (IC 50 ): 33.12 ?mol?L -1 and 18.35 ?mol?L -1 ; clinically relevant concentrations of propofol and midazolam reduced Na + peak currents by 27.66 % (P

To better understand the cleavage efficiency of muhiribozyme system on its RNA substrate in the presence and absence of divalent magnesium and monovalent sodium ions.we constructed pGEM-Coat'A,pGEM-Coat'A196Rz plasmids and pGEM-MDRl target plasmid.They were applied to transcribe RNAs with SP6/T7 transcription kit.Cleavage reactions were carried out in cell-free system and reaction products were analyzed by electrophoresis on 6% denaturing polyacrylamide gels in TBS buffer.The gels were dried and exposed to X-ray films for autoradiography.The Image J software was employed to analyze the dried gels.The results indicated that the cleavage efficiency of the muhiribozyme was dependent on the concentration of divalent Mg2+.The cleavage products increased with the concentrations of divalent Mg2+ and were Mg2+ concentration and time dependent.No cleavage product was obtained in the presence of lower than 200 mmol/L Na+ alone.On the contrary,monovalent Na+ inhibited the Mg2+ -induced cleavage reaction in Na+ and Mg2+ coexistance.The cleavage rate was significantly lower than that observed with divalent Mg2+ alone.These results suggested that divalent Mg2+ was required for muhiribozyme on substrate cleavage reaction in the physical condition,whereas monovalent Na+ was not.

Objective To investigate the effects of transplantatation human endothelial progenitor cells onneurological functional recovery from spinal cord injury model in rats,survival of transplanting cells and differentia-tion. Method The human endothelial progenitor cells were provided by Shanghai Developmental Biology Labora-tory.Forty SD rats were made for the animal model of spinal cord complete transection.Thirty survival SD rats wererandomly divided into three groups:sham operation gronp(group A, n = 10),operation/cell group(greup B, n =10) and operation/DMEM group (group C, n = 10).Suspension containing (hEPCs 6×106) was transplanted in-to the vertebral canal around injured spinal cord. In group C, equal volume of DMEM was injected insbead in the same way as in the group B. The BBB score was obtained 2,4,6,8 weeks after injection, Immunohistochemistry andin-situ hybridization were used to observe the cells survival and differentiation in the spinal cord. The BBB test wasperformed to study the functional improvement of cells. The SAS version. 1.3 software for statistics was to studyethology and functional improvement. The sum of ranks was checlced with Kruskal-Wallis Test and Nemenyi test.Results There were statistically significant differences in BBB scoring between group A and group B as well asgroup C after operation (P<0.05). The BBB score in group B was higher than that in group C after 2,4,6 and8 weeks,but lower than in group A. The hybridization in-situ and immunohistochemistry showed that transplantedcells survived for 8 weeks after transplantation and expressed specific characteristics for ancestral cell and differentiated into vascular endothelial cell (VEC). Conclusions After transplantation, hEPCs can survive, differenti-ate into vascular endothelial cell,and improve spinal cord function as compared with control group.

AIM: To observe the immunoregulation of all ogeneic cornea on the human peripheral blood T lymphocytes in vitro. METHODS: After Co-culture of human peripheral blood lymphocytes and allogeneic cornea in vitro, T lymphocytes were labeled by monoclonal ant ibody, and analyzed by fluorescent activated cell sorter (FACS). RESULTS: CD25 expression on T lymphocytes in control was 25.2%, after stimulated by the allogeneic cornea or PDB, CD25 expression on T lymphocy tes was 56.8% and 80.9%, respectively. After stimulated by the allogeneic corn ea, CD25 expression on CD 4+ or CD 8+ T lymphocytes were 67.3% and 52.3% , respectively. CONCLUSION: Allageneic cornea stimulates CD25 expression on huma n peripheral blood T lymphocytes, and the CD25 expression on CD 4+ T lymphocy tes is more prominent than CD 8+ T lymphocytes.

Objective To study the significance of combined detection of procalcitonin ( PCT ) , C-reactive protein( CRP) and lipopolysaccharide( LPS) for early diagnosis of bacterial infection in newborns. Methods Clinical data of ninety-eight newborns from neonatal ward of our hospital were retrospectively studied. Fifty cases with bacterial infectious diseases were selected as infection group,in the same period,48 cases with non-bacterial infectious diseases were selected as control group. In the 24 hours after admission before use of antibiotics,all of cases were picked blood used for testing CRP,PCT,LPS and blood culture, and the results were contrasted and analyzed. Results The levels of serum PCT,CRP and LPS in infection group were respectively significantly higher than those in control group,and the differences were statistically significant(P<0. 05,respectively). In gram-positive bacterium group,the positive rate of combined detection of serum PCT and CRP was obviously higher than that of single detection of PCT or CRP ( 91. 3% vs. 60. 9%,P<0. 05;91. 3% vs. 56. 5%,P<0. 05 respectively) . In gram-negative bacteria group,positive rate of combined detection of serum PCT and LPS was obviously higher than that of single detection of PCT or LPS respectively(88. 9% vs. 59. 3%,P<0. 05;88. 9% vs. 66. 7%,P<0. 05 respectively). Conclusion Joint detection can improve the diagnostic efficiency, and reduce missed diagnosis. And we can identify disease which predominantly infected by positive bacteria or negative bacteria through joint detection,which can contribute to the choice of clinical antibiotic drugs.