Objective To investigate the effects of case-based learning (CBL) applied in the clinical probation of anesthesiology.Methods Totally 313 undergraduates from the department of anesthesiology were randomly divided into 2 groups:CBL group (n =157) and LBL group (n =156).CBL methods were used to teach students in CBL group while traditional teaching methods were used to teach students in LBL group.After the causes being finished,a combination of theoretical examination and questionnaire were used to evaluate the teaching effectiveness between the two groups and a questionnaire was used to evaluate the recognition of teachers on CBL method.Data were managed by SPSS 17.0 software package.Quantitative data between groups were compared by t-test,enumeration data between groups were compared by chi-square test.Results Students in CBL group acquired higher score (86.7 ± 5.4) than those in LBL group (75.8 ± 4.5).82％ students in CBL group considered that their overall quality was improved while 42％ students in the LBL group thought so (P ＜ 0.05).75％ of faculties preferred the CBL methods.Conclusions Effect of CBL methods is better than traditional teaching methods and it's preferred by faculty and students.

Objective To investigate the changes in the number of microglias and astrocytes in the spinal dorsal born in a rat model of phantom limb pain.Methods Eleven healthy adult SD rats of both sexes weighing 290-300 g were randomly divided into 2 groups:sham operation group (group S,n =5 ) and unilateral sciatic nerve transection group (group SNT,n =6).Phantom limb pain model was induced by resection of a 0.5 cm segment of unilateral sciatic nerve in group SNT.In group S unilateral sciatic nerve was exposed but not transected.The animals were observed for autotomy and scored (0 =no autotomy,13 =the worst autotomy) after operation and were sacrificed on the 28th day after operation.The L5 segment of the spinal cord was removed for determination of the number of microglials (by iba-1 immuno-histochemistry) and astrocytes (by GFAP immuno-histochemistry).Results In group S no animal developed autotomy.In group SNT autotomy started from the 2nd day after operation and the score reached 9-11.Compared with group S,the number of the microglias and astrocytes in the spinal dorsal horn was significantly decreased in the operated side in group SNT ( P ＜ 0.05 ).Conclusion The number of microglias and astrocytes in the spinal dorsal horn is decreased in animals with phantom limb pain.

Objective To investigate the changes in the number of synapses and neurons in the spinal dorsal horn in a rat model of phantom limb pain. Methods Eleven healthy adult SD rats of both sexes weighing 209-300 g were randomly divided into 2 groups: sham operation group (group S, n = 5) and phantom limb pain group (group P, n = 6). Phantom limb pain was induced by resection of a 0.5 cm segment of unilateral sciatic nerve in group P. In group S unilateral sciatic nerve was exposed but not transected. The animals were observed for autotomy and scored (0 = no autotomy, 13 = the worst autotomy) after operation and were sacrificed on the 28th day after operation. The L3-6 segment of the spinal cord was removed for determination of the number of neurons (by Nissl's staining) and synapses (by synaptophysin immuno-histochemistry).Results In group S no animal developed autotomy. In group P autotomy started from the 2nd day after operation and the score reached 9-11. The number of the neurons in the spinal dorsal horn in all 4 segments and the number of synapses in L3 and 16 segments were comparable between the two sides and the 2 groups. The number of synapses in the spinal dorsal horn of L4and L5 segment was significantly larger in the operated side than in the contralateral side in group P. Conclusion The number of synapses in the spinal dorsal horn significantly increases in animals with plantom limb pain which induces no increase in the number of neurons in the spinal dorsal horn.

Objective To prepare anti-activin receptor-interacting protein 1 (ARIP1) polyclonal antibody.Methods The GST-ARIP1 fusion protein was expressed in E.coli BL21.A polyclonal antibody against ARIP1 was obtained by immunizing a rabbit with the purified GST-ARIP1 and the localization of mature ARIP1 protein in mouse brain tissues was detected by immunohistochemistry using the prepared anti-ARIP1 antibody.Results Anti-ARIP1 polyclonal antibody could bind specifically with recombinant ARIP1,but not recombinant ARIP2.Immunohistochemical staining showed that ARIP1 mainly expressed in hypothalamus and hippocampus using the prepared anti-ARIP1 antibody.Conclusion The polyclonal antibody against ARIP1 has been successfully prepared and can be used to do immunohistochemical analysis for ARIP1 protein expression.

Objective To detect the expression of matrix metalloproteinase-9 (MMP-9) gene in peripheral blood of patients with oral paraquat (PQ) poisoning and evaluate its predictive value for their prognosis.Methods Thirty-seven cases of oral PQ poisoning admitted to Linyi People's Hospital from January 2013 to June 2014 were enrolled,and they were divided into survival group (26 cases) and death group (11 cases) according to the survival sitnation in 28 days after poisoning;a healthy control group included 10 healthy people selected in the same period.The peripheral blood 3 mL was collected from each PQ patient on the 1st and 3rd day after admission,and in the healthy control group,3 mL peripheral venous blood was obtained under fast on the day for physical examination.The MMP-9 gene expression of peripheral blood mononuclear cells (PBMCs) was detected by reverse transcription-polymerase chain reaction (RT-PCR) methods;the serum MMP-9 concentration was determined by using enzyme-linked immunosorbent assay (ELISA);the serum PQ level was detected by high performance liquid chromatography (HPLC),and the amount of poison orally taken was recorded.The correlations between PQ amount orally taken,serum PQ level and MMP-9 expression were analyzed by Spearman correlation analysis.A receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of peripheral blood MMP-9 level for the 28-day prognosis of PQ poisoning patients.Results After admission the 1 day serum PQ level was (2.60 ± 1.29) mg/L,and the amount of poison taken was 50.0 (7.5,60.0) mL in the 37 patients with oral PQ poisoning.The MMP-9 gene expression level in PBMCs and serum MMP-9 protein level of both PQ poisoning groups were significantly higher than those of healthy control group,and the levels were gradually increased with the extension of poisoning time;the degrees of elevation in death group were more significant [the PBMCs' MMP-9 gene expression (A value):2.84± 1.16 vs.0.95 ± 0.23 on the 1st poisoning day,4.22± 1.75 vs.1.29 ±0.30 on the 3rd poisoning day;serum MMP-9 concentration (μg/L):2791.48± 1 230.88 vs.807.81±279.86 on the 1st poisoning day,4384.21 ± 1 781.97 vs.1 131.14±291.76 on the 3rd poisoning day,all P ＜ 0.05].Correlation analysis showed:there were significant positive correlations of oral PQ amount,serum PQ concentration to the MMP-9 gene expression in PBMCs and serum MMP-9 protein concentration in patients with oral PQ poisoning (all P =0.000).ROC curve analysis showed:the MMP-9 gene expression in PBMCs on the 1st day and the serum MMP-9 content on the 3rd day after admission had predictive value for 28-day prognosis in patients with oral PQ poisoning,and the ROC areas under the curve (AUC) was 0.820 and 0.776 respectively.When the cutoff value of MMP-9 gene expression level on the 1st day after admission was 0.90,the predictive sensitivity and specificity were 80.00％ and 63.64％ respectively;when the cutoff value of serum MMP-9 protein content on the 3rd day after admission was 904.36 μg/L,the predictive sensitivity and specificity were 80.00％ and 72.73％ respectively.Conclusion Oral PQ poisoning can lead to the MMP-9 gene expression in PBMCs and elevation of serum MMP-9 protein level in the body,and the MMP-9 gene expression has predictive value for the prognosis of patients with oral PQ poisoning.

Experimental course of pathology is a very important part of pathology teaching.Motivation of students was promoted through improving the professional knowledge of teachers,using teacher-student interacted teaching methods,increasing network courscs,using multimedia in teaching and adding extracurricular practice in our university.In the meantime,it was successful and fruitful to use diversified evaluation systems to assess the comprehensive experimental capacity of students,which can provide experiences in reform of pathological experimental teaching.

Objective To evaluate the effects of sepsis on pharmacodynamics of rocuronium in rats.MethodsForty SD male rats weighing 250-350 g were randomly divided into 3 groups: control group(group C,n =10) ; sham operation group(group S,n =10)and sepsis group (group Sep,n =20).Cerum was ligated and perforated to produce sepsis model in Sep group,rocuronium 3.81 mg/kg was intravenously injected at 6 or 16 h after operation,each time contains 10 rats.Cecum was not ligate and perforate in group S,but rocuronium 3.81 mg/kg was intravenously injected at 6 h after operation.Onset time,TOF no reaction period,duration of peak effect,clinical duration,totel duration,time for recovery of T1 to 10％,25％,50％,75％,90％ and recovery index were recorded by RM6240B signal acquisition and processing system.ResultsCompared with groups C and S,onset time was significantly prolonged,TOF no reaction period,duration of peak effect,clinical duration,total duration and time for recovery T1 to 10％,25％,50％,75％,90％ and recovery index were shortened in group Sep ( P ＜ 0.05).Onset time was significantly prolonged,time for recovery of T1 to 75％ was shortened when rocuronium injection at 16 h after operation as compared with that at 6 h after operation in group Sep( P ＜ 0.05).ConclusionSepsis can attenuate the effects of nondepolarizing neuromuscular blocker rocuronium,the degree is related to the stage of sepsis.

Objective To evaluate the effect of low-dose ketamine on the efficacy of electroconvulsive therapy (ECT) under propofol anesthesia in depressed rats. Methods Sixty adult male SD rats weighing 200-250 g were used in this study. The depression model was established by chronic unpredictable mild stress (CUMS). The animals were then randomly divided into 6 groups (n = 10 each): control group (group C), depression group (group D), propofol group ( group P), propofol + ECT group ( group PE), ketamine + propofol group ( group KP), and ketamine + propofol + ECT group (group KPE). Groups P and KP received intraperitoneal propofol 100 mg/kg and ketamine 10 mg/kg + propofol 80 mg/kg respectively, and groups PE and KPE received ECT after intraperitoneal injection of propofol 100 mg/kg and ketamine 10 mg/kg + propofol 80 mg/kg respectively once a day for 7 consecutive days. All rats underwent sucrose fluid consumption and Morris water maze tests before CUMS, after CUMS, and after treatment. Results Compared with group C, the sucrose consumption percentage was significantly decreased, the escape latency was prolonged, and the time spent in the target quadrant (the original platform quadrant) was shortened after CUMS in D, P, PE, KP and KE groups ( P ＜ 0.05). Compared with group D,the sucrose consumption percentage was significantly increased (P ＜ 0.05), while no significant change in the escape latency and time spent in the target quadrant was found after treatment in group KPE ( P ＞ 0.05 ), and the sucrose consumption percentage was significantly increased, the escape latency was prolonged, and the time spent in the target quadrant was shortened after treatment in group PE ( P ＜ 0.05). Compared with group PE, the sucrose consumption percentage was significantly increased, the escape latency was shortened, and the time spent in the target quadrant was prolonged after treatment in group KPE ( P ＜ 0.05). Conclusion Low-dose ketamine can not only enhance the efficacy of ECT under propofol anesthesia in depressed rats, but also reduce cognitive impairment induced by ECT.

Objective: To assess the expression of retinoic acid receptor beta 2 (RAR-β2) in breast tumor and to evaluate the relationship between RAR-β2 expression and tumorigenesis of breast cancer. Methods: Immu-nohistochemistry was used to detect the expression of RAR-β2 protein in specimens from 40 cases of breast cancer, 40 cases of atypical ductal hyperplasia, 40 cases of fibroadenoma and 20 cases of normal breast tissues. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of RAR-β2 gene in 20 cases of breast cancer, 20 cases of atypical ductal hyperplasia, 20 cases of fibroadenoma, and 20 cases of normal breast tissues. Results: Immunohistochemical staining results revealed that the positive expression of RAR-β2 protein showed nuclear staining. The positive expression rate of RAR-β2 was 30% (12/40) in breast cancer, 17.5% (7/40) in atypical ductal hyperplasia, 87.5% (35/40)in fibroadenoma, and 95% (19/20) in normal breast tissues. The expression of RAR-β2 protein in breast cancer was significantly lower than that in normal breast tissues (X~2=26.30, P<0.001). The expression of RAR-β2 was not significantly different between atypical ductal hyperplasia and breast cancer (P>0.05). No correlation was found between the expression of RAR-β 2 protein and the tumor size, menopausal age, lymph node metastasis, clinical stage, histological grade, or protein expression of ER and PR in breast cancer tissues (P>0.05). Follow-up results showed that 3 out of 28 patients with negative RAR-β2 expression had visceral organ metastasis, but only one of the 12 RAR-β2 positive patients had osseous metastasis. RT-PCR analysis showed that the positive expression rate of RAR-β2 mRNA in breast cancer, atypical ductal hyperplasia, fibroadenoma and normal breast tissues was 25% (5/20), 35% (7/20), 85% (17/20) and 100% (20/20), respectively. The RAR-β2 mRNA expression rate in breast cancer was significantly lower than that in normal breast tissues (X~2=30.43, P<0.001). No significant difference in RAR-β2 mRNA expression was found between atypical ductal hyperplasia and breast cancer tissues (P>0.05). Conclusion: RAR-β2 gene may play a repressive role in the initiation of breast cancer, and the loss of the expression of RAR-β2 gene may be the initial step in breast carcinogenesis.

Objective To investigate the distribution of eosinphililic neurons ( ENs),reactive astrocytes ( RAs),and infarction after transient cerebal ischemia,and the time profile of pathomorphological changes.Methods Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10 minutes unilateral common carotid artery occlusions with a 5 hours interval.Laser Doppler flowmetry was used to detect intra-ischemic anterior cortex blood flow.Animals were sacrificed at 24 hours,4 days,2 weeks,4 weeks,16 weeks and the brain were prepared for pathomorphological assay.Results Intra-ischemic laser Doppler flowmetry show significant ischemia during carotid artery occlusion:22.1％ ± 9.5％,26.3％ ± 4.9％,37.5％ ± 3.5％,F =67.219,P ＜ 0.01 ; the decrease was significantly greater in the anterior cortex.ENs appeared in middle and deep layers at 24 hours postischemia,and ENs area extend to all layers of cortex by 4 days.Large areas of high EN density ( ≥80/mm2) evolved to infarcts between 4 days and 4 weeks.Posterior cortex evolved to low EN area ( ＜ 80/mm2) without transformation into infarcts.RAs were consistently distributed in areas with ENs,and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks. Delayed astrocytic death took place in the RA areas with high EN density.Conclusion Density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue.