Background Identifying and testing of pain is very necessary for the care and decrease of the suffering of experimental animal in medical experiment.Effective method for testing the pain and distress status of experimental animal with eye disease is still absent in China.Objective This pilot study was to establish an evaluating system for assessing the pain and distress status of bullous keratopathy rabbit model.Methods This study was approved by the Ethic Committee of Beijing University First Hospital.Twelve healthy New Zealand rabbits were selected in this experiment.Bullous keratopathy model was established in the left eyes of 9 rabbits by scraping corneal endothelium as the experimental team,and other 3 rabbits were served as the control team.The cornea lesion was examined by manipulate slit lamp,and the central cornea thickness (CCT) was measured by ultrasound biomicroscopy (UBM).Weight+20 Indexes For Pain and Distress Status Referring Guidelines for Pain and Distress in Laboratory Animals made by International Animal Care and Use Committee (IACUC) were assessed and measured as well.Results Corneal edema and opacity were obvious 1 day after surgery.Corneal bullous appeared from the third day after surgery,and cornea erosion was seen at the location of bullous breakage.The corneal lesions remained until 14 days after surgery.CCT value was (1468±100),(2313±588),(2391±271) and (2362±151) μm,respectively in day 1,3,7 and 14 after the establishment of models,which showed significant increase in comparison with the preoperative CCT (390±6)μm (all P=0.000).However,no significant difference was seen in the CCT between day 3,7 and 14 (P＞0.05).Body weight of the rabbits was (3.29±0.20),(3.20-0.17),(2.77±0.25) and (3.10±0.30)kg respectively in day 1,3,7 and 14 after operation,with significant decrease in comparison with the pre-operative weight (3.52-0.18)kg in the experimental team (P=0.008,0.007,0.003,0.004).The scores for pain and distress status of all rabbits in pre-operation of the experimental team and in the control team were zero,and the score was 7 (7,7),11 (10,12),9 (8,10),9 (9,9)in day 1,3,7 and 14 in the experimental team after surgery,with the highest score in day 3,which was bullous and bullous breakage duration.Seven of twenty indexes,including the reduce of diet and drinking,self-imposed isolation/hiding,grinding teeth,aggression,deceased movement,abnormal posture,vocalization occurred in the model animals after surgery.Conclusions Weight+20 Indexes For Pain and Distress Status is an effective,impersonal and quantitative method for observing and evaluating the pain and stress status in bullous keratopathy rabbit.

Objective To explore the effects of ephedrine on the hippocampal cell apoptosis and behavioral performance after hypoxia-ischemia brain injury in neonatal rats.Methods Totally 90 7-day rats were randomly divided into 3 groups,ephedrine treatment group,model group,and sham group.Hypoxia-ischemia brain injury model was established by permanently ligating right common carotid artery.Ephedrine (1.5 mg/kg,once per day) was injected intraperitoneally to the rats of ephedrine treatment group for 7 d,and the rats of model group was given normal saline at the same volume.At the following time interval of 6 and 12 h,and 1,3,and 7 d after hypoxia,the expression of bcl-2 and bax were detected in the hippocampal region by immunohistochemical staining.At 4 weeks after surgery,behavioral changes in the remaining rats were tested by Morris water maze.Results Compared with model group,the expression of bcl-2 in the ephedrine treatment group was significantly increased after hypoxic-ischemic injury,peaked at 1 d and decreased in 3 d after operation.And the expression of bax in the ephedrine treatment group was decreased in 1 d after hypoxic-ischemia.The average time of escape latency was gradually decreased in each group.However,from the 3rd to 5th day,it was much shorter in ephedrine treatment group than in model group.In addition,the frequency platform passing in the ephedrine treatment group and the percentage of swimming distance traveled in the previous target quadrant was significantly greater than those of the control group.Conclusion Ephedrine upregulates bcl-2 and downregulates bax in the hippocampus of neonatal rats after hypoxia-ischemia,and improves their ability of learning and memory.

Humans ; Pancreatic Neoplasms ; prevention & control ; therapy

China ; Congresses as Topic ; Humans ; Pharyngeal Diseases ; Tracheal Diseases

OBJECTIVETo observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.

METHODSWe treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.

RESULTSCompared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).

CONCLUSIONGefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; Leydig Cell Tumor ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Proteins ; metabolism ; Neoplasms, Germ Cell and Embryonal ; drug therapy ; metabolism ; pathology ; Quinazolines ; pharmacology ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

Objective To understand acute rejection differences between untreated recipients and rapmycin-treated recipients in a rat free flap allotransplantation model. Methods Brown groin free flaps were transplanted to Lewis recipients. In the treated group, recipients were treated with rapamycin at the dose of 4 mg/kg every day from day 0 to day 14 after transplantation. In the untreated group, recipients didn't receive any treatment. Allografts were evaluated clinically and histologically. Results Allografts in the treated group showed epidermolysis as sign of rejection.Rejection sign of untreated grafts was ischemic necrosis of whole skin. In histological evaluation, the treated grafts showed "band-like" lymphocytes infiltration in the upper dermis when rejection occurred, while the untreated grafts showed thrombosis in the subdermal vessels. Conclusion The differences between the two groups implied that embolization may be responsible for the rejection of free flap allotransplantation in rat model.

Objective To identify the inclusion compound of ginsenoside Rh2 with hydroxylpropyl-?-cyclodextrin (HP-?-CD) in aqueous solution, and to investigate the change of thermodynamic parameters during the inclusion process. Methods The measurements of the inclusion mechanism, inclusion molar ratio of the host and guest molecules, and change of thermodynamic parameters were carried out by the following methods separately; thin layer chromatography (TLC), differential scanning calorimetry (OSC), infrared (IR) spectrometry, phase solubility method, and thermodynamic method, respectively. Results That all the phase solubility diagrams showed a typical AL-type in various temperatures and suggested the formation of a soluble complex of 1∶1 molar ratio. TLC and IR Spectroscopy confirmed that the significant interaction between the host and guest molecules was probably due to the inclusion of aglycone of Rh2 molecule into the hydrophobic cavity of HP-?-CD molecule. The change in the thermodynamic parameters suggested that the inclusion could proceed spontaneously (?G

Objective To investigate the diagnostic and predictive value of measuring peritoneal inflammatory cytokines in predicting biliary fistulas in patients undergoing biliary surgery.Methods Peritoneal samples and serum samples of 3227 biliary surgery patients were collected on the first,third,fifth postoperative day,and IL-6,IL-8,IL-10,TNF-? and CRP of the samples were measured.Patients were divided into two groups:those with clinical evidence of biliary fistulas and those without any evidence of biliary fistulas.The age,sex,operative method and operative time between the two groups were compared.Results There was a negative correlation between biliary fistulas and age,sex and operation mode;but blood loss,operation time and common bile duct diameter had positive correlation with biliary fistulas.Peritoneal cytokine levels were significantly higher in patients with biliary fistulas as compared to those without biliary fistulas.Conclusions The peritoneal cytokine level is a sensitive parameter of peritoneal inflammation and can be as an additional diagnostic tool for the early prediction of biliary fistulas after biliary surgery.

Objective To investigate the calculation method and its inlfuencing factors of Z scores in the aortic root diameters measured by echocardiography in children. Methods A total of 105 children with median age 19 months, who came to Shenzhen Children′s Hospital from March 2012 to October 2012 were included. The diameters of aortic ring (ARD) and aortic sinus (ASD) were measured by two dimension echocardiography, Z scores of ARD and ASD were calculated using two different normal reference values regression equation and mean square error derived from Shenzhen children′s hospital (C method) and Pettersen et al (P method). The regression equation from C method and body surface area (BSA) formula from P method were adopted to calculate Z scores of ARD and ASD (ZH method). The Z results of ARD and ASD calculated by those three methods were compared and were analyzed for their normality probability distributions. Results Z scores of ARD and ASD derived from C method were all showed as normal distribution (P=0.067 and 0.650). Z scores of ARD and ASD derived from P method were all showed as normal distribution (P=0.208 and 0.970). Z score of ARD derived from ZH method was showed as non-normal distribution (P=0.027), but Z score of ASD was normal distribution (P=0.430). There were no significant differences in ARD-Z calculated by C method (0.41±0.89), P method (0.23±0.85) and ZH method (0.36±0.94) (F=1.117, P=0.309). There were signiifcant differences in the Z scores of ASD calculated by C method (0.38±0.89), P method (0.58±0.71) and ZH method (0.36±0.84) (F=5.443, P=0.005). Z scores of ARD (r=0.917, P=0.000) and ASD (r=0.900, P=0.000) calculated by C method correlated well with that by P method. Conclusions Calculation method of BSA and normal reference values regression equation were the main influencing factors of Z score value in quantifying children aortic root diameters by echocardiography. For the clinical applications. The normal reference value should be used which is suitable for the Chinese children.

Objective To establish a calcification mode in vitro of human vascular smooth muscle cells (HVSMCs) induced by β-GP. Methods Primary HVSMCs were obtained from human embryo by plant method and confirmed by stain with α-sm-actin antibody. The cells after 4-6 passages were divided into two groups.The control group was incubated with normal DMEM medium while the calcification group was incubated with the medium containing 10 mmol/L β-glycerophosphate for 10 days.Calcification was confirmed by alizarin red S stain and alkaline phosphatase(ALP) assays. Results The primary cells observed by S-P stain were positive and the cells after being stained were pale yellow. After being induced with β-GP, the cells of calcification group began concentric growth and formed vesicles. Alizarin red S staining showed that the reaction of calcifying nodules was red,ALP activity was higher than that of controls at various time points(4 d,6 d,8 d and 10 d,P＜0.01 ).Conclusion The HVSMCs could be induced into calcification in vitro by β-GP, and this model contribates to further studies of vascular diseases.