BACKGROUND: It is demonstrated that ropivacaine has less toxicity than bupivacaine, but bupivacaine has higher liposolubility and efficacy, so a less dose of bupivacaine is needed in clinical comparing with ropivacaine. Serious convulsion is usually followed by cardiotoxicity induced by local anesthetics. The ratio of medial lethal dose (CD50) and median convulsant doses (LD50) is usually used to assess the comparative safety of local anesthetics.OBJECTIVE: To establish CD50 and LD50 of 2% lidocaine, 0.75% bupivacaine and 0.75% ropivacaine for Kunming mice and select proper indicator for neurotoxicity, then to compare neurotoxicity of the three local anesthetics on central nervous system.DESIGN: Randomized and controlled study.SETTING: Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: This study was carried out from July to December in 2002 in the Laboratory of Anesthesiology, Tongji Hospital, Tonji Medical College, Huazhong University of Science and Technology. Totally 310Kunming mice aged of 1-month with clean grade were enrolled in this study.METHODS: ① To determine the relation of dose of local anesthetics with conclusion rate and death rate in mice. Todetermine the dose-effect relationship for ropivacaine, 50 mice were selected and divided into 5 groups with 10rates in each group who received dose of 76.80, 68.69, 61.44,49.15, 31.46 mg/kg respectively. For bupivacaine, 90 mice were divided into 9 groups, with 10 rates in each group who received intraperitoneal dose of 50.00, 47.29, 44.72, 42.29, 40.00, 35.78, 32.00, 28.62, 25.60 mg/kg respectively. For lidocaine, 100 mice were divided into 10 groups,with10rates in each group who received dose of 183.11, 163.77, 146.48,131.02, 117.19, 93.75, 75.00, 60.00, 48.00, 38.40 mg/kg respectively. For each local anes thetic, the rates of convulsion or death were tried to distribute on both sides of 50% symmetrically. On the dose-response curve, 4or 5 well-spaced points were obtained for probit analysis to determine CD50 and LD50 of each agent. ② The effect of different dose of lidocaine on conclusion duration and c-fos expression in brain with different doses.Forty mice were divided into 4 groups with 10 rates in each group who received 0, 30%, 60% and 90% convulsant doses of lidocaine intraperitoneally. The duration of convulsion were recorded carefully for the convulsant mice that should be marked correctly for next procedure. Two hours later, the convulsant mice were anesthetized deeply and fixed by transcardiac perfusion for Immunohistochemistry to detect c-Fos expression. ③ Comparison of neurotoxicity induced by CD50 of three agents.Thirty mice were randomly divided into 3 groups with 10 rates in each group who received intraperitoneally CD50 of lidocaine, bupivacaine and ropivacaine respectively. The duration of convulsion and the number of neurons expressed with c-Fos in mice brain were compared among these three groups.MAIN OUTCOME MEASURES: The response of mice to intraperitoneal local anesthestics, duration of convusion and c-Fos expression using immunohistochemistry methods.RESULTS: Date of totally 310 mice was entered into final results analysis. ① The relation of dose of local anesthetics and conclusion rate or death rate in mice. The therapeutic index (LD50/CD50) of 2% lidocaine,0.75% bupivacaine and 0.75% ropivacaine were 2.89, 1.48 and 1.34, respectively. ② c-Fos expression induced by lidocaine in mice brain: The cFos expression in mice brain was mainly distributed in three zones-thalamencephal, hypothalamus, amydyla and pyriform cortex. ③ Compare of the duration of convulsion and number of neurons with c-Fos expression induced by different dose oflidocaine. Compared with control group, the duration of convulsion and number of neurons with c-Fos expression in amydyla and pyriform cortex all increased significantly in CD30, CD60and CD90 group (P ＜ 0.05). ④ Neurotoxicity induced by CD50 of lidoacaine, bupivacaine and ropivacasine The duration of convulsion and expression of c-fos in amydyla and pyriform cortex were significantly increased in ropivacaine group compared to bupivacaine or lidocaine group intraperitoneally (P ＜ 0.05). There were no significant differences in the duration of convulsion and expression of c-Fos between lidocaine and bupivacaine group (P ＞ 0.05).CONCLUSION: Compared with bupivacaine, ropivacaine produced less toxicity when identical dose was used in clinic. It is indicated that if an accidental convulsion induced by ropivacaine, it may be more severe than that induced by correspondent either lidocaine or bupivacaine. It may be the reason that ropivacaine have less lipid solubility, absorbed easily from this tissue compartment, and to get a high concentration in blood.

Objective To detect the level of IL-10 and IL-21 in serum of patients with psoriasis and explore its significance of judge the extent of serious condition and treatment of psoriasis.Methods According to clinical dermatology edited by Zhao Bian diagnostic criteria,selected 65 cases of patients with psoriasis during the period from March to September in 2014 ad-mitted to Department of Dermatology of out-patient and hospitalization.25 cases were progressive stage (group A),20 pa-tients with stable stage (group B),20 patients were in recovery period (group C),and 25 cases of control group (group D). With ELISA method detect the expression level of IL-10 and IL-21 in serum of each group respectively.PASI score will take to all psoriasis patients meanwhile.Results The expressionlevels of IL-21 in serum of A (127.59 ± 16.09 pg/ml),B (105.74±21.08 pg/ml)two groups were significantly higher than that in D group (85.46±14.25 pg/ml),and the differ-ence were statistically significant (t=5.174,4.863,both P <0.01),while the expression levels of IL-10 in A (10.64±3.23 pg/ml)and B (12.27±2.18 pg/ml)were both lower than D group (20.29±2.51 pg/ml),and the difference were statisti-cally significant (t=2.031,2.027,both P <0.05).The expression of IL-21 (94.03±8.90 pg/ml)in C group was higher than that in D group,and the difference was statistically significant (t=2.033,P <0.05).The level of IL-21 in A group was higher than that in group C,and the difference was statistically significant (t=2.352,P <0.05),while the expression level of IL-10 was lower than that in C group (19.69±1.54 pg/ml),and the difference was statistically significant (t=2.071,P<0.05).The expression level of IL-21 in the serum of patients with psoriasis and PASI score was positively correlated (r=0.508,P =0.027),while the expression level of IL-10 and PASI score was negatively correlated (r=-0.413,P =0.039). Conclusion The progression of psoriasis is related to the increasion of IL-21 or the decreasion of IL-10.The detection of IL-21 and IL-10 was beneficial to judge the severity and efficacy evaluation of psoriasis.

BACKGROUND:Tropomyosin 4 level has been found to be an increase in the spinal cord based on the 2-DE/MALDI-TOF/MS method. However, there is little report about the relationship between tropomyosin 4 and pathogenesis and progress of spinal cord injuries.
METHODS/DESIGN:Randomized control ed trial:rat models of complete spinal cord transection were made and expression levels of tropomyosin 4 at 3-28 days after modeling were determined by two-dimensional electrophoresis, animo acid serie analysis, quantitative PCR and western blot. Experiment for exporing the genetic mechanism:effects of tropomyosin 4 scilencing by lentivirus recomnination technology on the dendrite length of spinal cord neurons in vitro were observed, and its effects on the neurological function of rats after complete spinal cord transaction were assessed through Basso, Beattie, and Bresnahan scoring.
DISCUSSION:This study wil be powered to provide a novel and effective treatment strategy for neurological function recovery after spinal cord transection based on the lentivirus recomnination carrying tropomyosin 4, as wel as optimistic future for clinical gene treatment of complete spinal cord transaction through figuring out the underlying mechanism.
ETHICAL APPROVAL:This study was approved by the Ethics Committee of Kunming Medical University, China. The surgical operation and postoperative care of rats were in line with the rules of Chinese Experimental Animal Protection and Ethics Committee, and the guideline of the National Institutes of Health

Objective To investigate the influences of imatinib on Survivin gene expression in bcr-abl-transformed leukemia cells.Methods Firstly,PCR and Western blot were carried out to detected Survivin expression with imatinib treatment in 32Dcl3 and 32D-bcr-abl cell lines.Then the luciferase reporter plasmids containing human Survivin promoter as well as its deletion and site-directed mutation were constructed to identify the essential responsive elements for suppressing Survivin promoter activity by imatinib.Chromatin immunoprecipitation was performed to confirm the binding of c-myc to Survivin promoter.10058-F4,a small molecule c-myc inhibitor,was used to disrupt c-myc activity and evaluate its anti-leukemic effect combined with imatinib.Results Both of mRNA and protein level of Survivin in bcr-abl-transformed cells were downregulated upon imatinib treatment.The decrease of Survivin expression was controlled at the transcriptional level through a mechanism in which imatinib repressed survivin promoter activity by disturbing the interaction between c-myc and E-box elements.Interruption of c-myc activity by 10058-F4 exerted an anti-leukemia effect with enhancing the sensibility of K562/G01 cells to imatinib.Conclusion Imatinib down-regulates Survivin expression through c-myc-mediated transcription and interference with c-myc might be a potential utility for treatment of imatinib resistant leukemia.

Objective To observe the effect of sedative drugs applied on uncooperative children in CT examination. Methods 204 samples were divided into groups of baby, infant and preschool children. 20 minutes before CT scan, chloral hydrate and diazepam were taken by those uncooperative children seperately. Drug dosage was calculated according to body weight (kg). Results The effective rate of chloral hydrate were 96.8%(30 cases) in baby group,54.2%(13 cases) in infant group and 53.9%(21eases) in preschool children group, while that of diazepam were 100%(9 cases), 82.6%(19 cases) and 87.2%(68 cases) respectively.Conclusion When approriate sedative drugs were taken by uncooperative children, satisfactory scanning images can be obtained in CT examination

Objective To discuss the value of radiation dose reduction with directed digital radiography application in chest X-ray. Methods Patients were in posterior-anterior standing posture for test. To compare DR exposure parameters with traditional roentgenography in the same exposure condition of 125 kVp, automatic mAs, 180cm focus-film distance. Patients' thoracic thickness was 22-25cm. Results DR can reduce exposure dose by 44.2%. Conclusion Directed digital radiography can improve the quality of chest images,make working flow more reasonable, enhance the working efficiency and reduce patient's x-ray dose as well.

Objective To investigate the effect of midazolam on human sperm motility in vitro.Methods Sperm samples were obtained from normal adults and prepared with discontinuous percoll gradient centrifugation technique.The samples were randomly divided into 3 groups (n =10 each):control group and 2 midazolam groups.The samples were incubated with normal saline in control group and with midazolam with the final concentrations of 5 or 1 μg/ml in 2 midazolam groups.The samples were incubated for 60 min in an airtight container at 37 ℃.Then human sperm motility was examined in vitro at 37 ℃ and analyzed by the computer-assisted sperm analysis at 10,30 and 60 min exposure to midazolam,including sperm motility (a + b)％,curvilinear velocity,straight line velocity,average path velocity,amplitude of lateral head displacement,beat-cross frequency,linearity,wobble,straightness,and mean angular displacement.Results There was no significant difference in the parameters of human sperm motility within each group and between groups (P ＞ 0.05).Conclusion Midazolam has no significant effect on human sperm motility in vitro.

Objective To investigate the effect of melatonin on ketamine-induced apoptosis in hippocampal neurons of fetal rats.Methods Sixteen to eighteen day pregnant Sprague Dawley rats were anesthetized.The fetal rats were obtained under sterile condition and decapitated.The hippocampal neurons were isolated and primary cultured for 5 days.The primary cultured neurons were randomly divided into 5 groups (n =6 each):control group (group C),ketamine group (group K),and 1.0,2.5 and 5.0 mmol/L melatonin groups (groups M1-3 respectively).Ketamine with the final concentration of 1 000 μmol/L was added to the culture medium and the neurons were incubated for 3 h in group K.In groups M1-3,1.0,2.5 and 5.0 mmol/L melatonin were added to the culture medium,respectively,at 60 min before the addition of ketamine,and the neurons were incubated for 3 h.While the equal volume of normal saline was added instead in group C.The neuronal viability during the developmental phase was assessed by MTT assay.The mitochondrial membrane potential (Ψm) was measured by flow cytometry.The expression of cAMP response element binding protein phosphorylation (p-CREB (Ser133)),Bcl-2,Bax,and cytochrome C was detected by Western blot.Results Compared with group C,the neuronal viability and Ψm were significantly decreased,and the expression of p-CREB and Bcl-2 was down-regulated,while the expression of Bax and cytochrome C was up-regulated in group K (P ＜ 0.05).Compared with group K,Ψm was significantly increased in groups M2 and M3,and the neuronal viability was significantly increased,the expression of Bcl-2 was up-regulated,while the expression of Bax and cytochrome C was down-regulated in groups M1-3 (P ＜ 0.05).Conclusion Melatonin can protect the hippocampal neurons of fetal rats from apoptosis triggered by ketamine via regulating the expression of Bcl-2 and Bax,stabilizing Ψm,inhibiting the release of cytochrome C from mitoehondria,and preventing apoptosome formation.

Objective To investigate the effects of isoflurane anenthesia on myocyte enhancer factor 2(MEF2) signaling pathway in neonatal rat hippocampus. Methods Twenty-four 5-day-old SD rats of both sexes,weighing 10-13 g, were randomly divided into 2 groups ( n = 12 each): control group (group C) and isoflurane group (group I). In group I, 1.5% isoflurane in 100% O2 was inhaled for 6 h. Group C received no treatment.Three rata in each group were sacrificed at 2, 4, 6 h of isoflurane anenthesia and 24 h after isoflurane anenthesia (T1-4), and the hippocampi removed for determination of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsinⅠ mRNA expression (by PT-PCR) and synapsin Ⅰ protein expression (by Western blot).Results Compared with group C, the expression of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsin Ⅰ mRNA at T1-3 and synapsin Ⅰ protein at T2-4 was up-regulated in group I ( P ＜ 0.05). Conclusion Inhalation of anaesthetic concentration of isoflurane may affect synapse formation during the development of central nervous system by actirating hippocampal MEF2 signaling pathways in neonatal rats.

Objective:To observe the expression of interleukin-17 and the infiltration of eosinophilic cells in nasal polyps and allergic rhinitis, and investigate the roles of IL-17 and eosinophils in the etiology of nasal polyps and allergic rhinitis.Method:A study was conducted on 21 patients of nasal polyps, 18 ones of allergic rhinitis and 12 normal individuals. Immunohistochemical stain with the rabbit monoclonal antibodies of IL-17 was carried out.The eosinophilic cells infiltrated in different tissues were stained with HE, then counted under high power filed.The data was analyzed with ANOVA of SPSS12.0 software.Result:Many IL-17 stained cells were found in the samples of nasal polyps and allergic rhinitis, which were significantly higher than those in normal individuals(P<0.05). Positive cell number in tissues of allergic rhinitis was similar to that in nasal polyps, but higher than in normal individuals. As for HE staining, there was no significant deviation of numbers of eosinophilic cell in tissue between allergic rhinitis and nasal polyps,while which differed from the normal ones(P<0.05).Conclusions:①IL-17 is a newly cytokine which expressed in mucosa of allergic rhinitis and nasal polyps tissue. It indicates the degree of immunological reaction and inflammatory reaction, and can be used as an index to research the mechanism of nasal polyps as well as allergic rhinitis.②The eosinophilic cells count was correlated with the amount of IL-17 positive cells in nasal ployps and with allergic rhinitis correlation coefficients were R=0.606(P<0.01)and R=0.446(P<0.05) respectively . It seems that eosinophils, which are regulated by IL-17, play an important roles in the development of nasal polyps and allergic rhinitis.