BACKGROUND: Caspase-3 is a cysteine proteinase that can promote cell apoptosis. Ciliary neurotrophic factor has many kinds of biological activities, such as protecting various neurons from injury, especially motorial neurons, thereby reducing the occurrence of nerve cell injury.OBJECTIVE: To observe the Influence of ciliary neurotrophic factor on spinal cord caspase-3 expression in different ages rats following sciatic nerve injury, aiming to make further investigation about the changing regularity and modulating character of peripheral nerve damage at various ages rats.DESIGN: Randomized controlled experiment.SETTING: Ultrasound Department and the 5th Research Institute of Daping Hospital, Field Surgery Research Institute of the 3rd Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out at the Field Surgery Research Institute of the 3rd Military Medical University of Chinese PLA from April 2000 to December 2001. Altogether 810 wistar rats including infant rats with body mass of 30-100 g (birth age of 20 days), adult rats of 200-350 g (birth age of 4 m), elder rats of 400-800 g (birth age of 18-24 m),were selected with 270 rats in each age stage. Female and male does not limit.METHODS: [1] Experimental animal grouping: Various age rats were randomly divided into normal group (n=18), ciliary neurotrophic factor group(n=126) and physiological saline group (n=126), rats in ciliary neurotrophic factor group and physiological saline group were subdivided into 1 day, 3days, 1 week, 2 weeks, 4 weeks, 8 weeks, 12 weeks subgroups. [2] Animal model preparation: In Ciliary neurotrophic factor group and physiological saline group, rats were cut off a piece of sciatic nerve of 2 mm long, both ends connected with silica tube for constructing neuranagenesis cab, in which 15 μL recombined ciliary neurotrophic factor (25 mg/L) was injected in ciliary neurotrophic factor group, and 15 μL physiological saline in physiological saline group. [3] Preparation and examination of specimen:Six rats were randomly selected from each group, lumbar L3-5 spinal cord were obtained for carryingonnation, caspase-3 activity examination and Western blotting examination.MAIN OUTCOME MEASURES: [1] Expression of spinal cord caspase-3 with IHC assay. [2] Distribution and expression intensity of spinal cordcaspase-3 by Western blotting technique. [3] Changes in caspase-3 activity.RESULTS: Altogether 810 rats completed the experiment, all data was entered the final result analysis. [1] Expression of spinal cord caspase-3 with IHC assay: After injury, neuronal caspase-3 expression became strengthened at injured side of various age physiological saline groups, which obviously increased at posttraumatic 2 weeks and 4 weeks, but less expressed at 8 weeks and 12 weeks; while in ciliary neurotrophic factor group, posttraumatic caspase-3 expression was mostly obvious at 2 weeks and 4 weeks. [2] Distribution and expression intensity of spinal cordcaspase-3 by Western blotting technique: Caspase-3 expression was not significant difference between various age subgroups in normal group(P ＞ 0.05). Comparing to the same age normal group and non traumatic group, caspase-3 expression was strengthened at 1 week, 2 weeks, 4 weeks in various age physiological saline group and ciliary neurotrophic factor group damages (P ＜ 0.05-0.01); in ciliary neurotrophic factor group, caspase-3 showed weaker expression at 1 week, 2 weeks, 4 weeks in infants; 2weeks and 4 weeks in adults, 4 weeks in elders comparing to corresponding physiological saline group, difference was significant (P ＜ 0.05-0.01).The comparison between untraumatic side of each group and normal group showed no statistical difference (P ＞ 0.05). [3] Changes in caspase-3 activity: In child, adult and elder physiological saline groups, caspase-3 activity was increased in traumatic spinal cord, moreover caspase-3 activity was higher than elder and adult group at various age point (P ＜ 0.05-0.01); in child, adult and elder ciliary neurotrophic factor groups, caspase-3 activity is lower than physiological saline group (P ＜ 0.05-0.01). After damage,caspase-3 activity at traumatic side in physiological saline group and ciliary neurotrophic factor group was difference from normal group but without significant meaning (P ＞ 0.05), except the expression in child posttraumatic 12 weeks group was lower than normal group (P ＜ 0.05).CONCLUSION: After sciatic nerve damage, caspase-3 protein expression and activity were found to be increased in spinal cord of different age groups rats which can be reduced by extragenous ciliary neurotrophic factor, which playing protective role on spinal cord nerve cells, such protection would gradually attenuated in child group, adult group to elder group in turns.

This paper introduces a novel animal behavior monitoring system, in which a acoustooptic shielded room, computer multimedia technology, image acquisition technology, signal analysis technology are involved. This system can implement long-term non-interference automatic monitoring of the experimental animal with features of accurate recording, complete and objective analysis, and convenient and effective statistics. Applied widely to the animal model for post -traumatic stress disorder (PTSD), the system is designed to be used for other biomedical studies.

ObjectiveTo evaluate the reliability and validity of the of Disability Assessment Schedule Ⅱ of World Health Organization(WHO-DASⅡ) applied in assessment of the function of disabled athletes.Methods56 disabled people,26 wheelchair basketball athletes and 30 patients,were sampled and completed WHO-DASⅡ questionnaire.The test and re-test had been administrated in 10 days.Barthel Index(BI) and Sports Classification Test had been implemented for the functioning assessment.ResultsThe test-retest reliability for WHO-DASⅡ in six dimensions were: 0.978,0.807,0.938,0.877,0.998,0.935,0.986 and all were in very significant level(P<0.01).There were negative correlation between WHO-DASⅡ scores and BI in total and in every dimension,the correlation coefficients were:-0.77,-0.17,-0.71,-0.82,-0.33,-0.73,-0.61.The correlation coefficients between WHO-DASⅡ total and six dimensional scores and the grade of sports classification were:-0.741,-0.378,-0.806,(-0.541),-0.293,-0.510,-0.677.ConclusionWHO-DASⅡ can be used in assessing the function of disabled athletes.

Objective To investigate the feasibility of transfecting gene into HepG2 by therapeutic ultrasound combined with microbubble,and to explore the optimal ultrasonic parameters for high transfection efficiency.Methods HepG2 cells were cross-interval planted in 24-well plates.EGFP plasmid DNA and microbubbles were added to the cultured HepG2 and three parameters of the therapeutic ultrasound were optimized.Firstly,set ultrasonic intensity gradient (group A,from A1 to A5),which were 0.4 W/cm,0.8 W/cm2,1.2 W/cm2,1.6 W/cm2 and 2.0 W/cm2 respectively.Secondly,set the duty cycle gradient group (group B,from B1 to B3),which were 10％,20％ and 50％ respectively.Lastly,set the exposure time group(group C,from C1 to C3),which were 30 s,90 s and 180 s.After 24 hours,the expression of GFP was observed by fluorescence microscopy,the transfection rates was detected by flow cytometry,the cell death were assessed by trypan blue exclusion.Results The plasmid transfection efficiency was varied under different ultrasonic parameters.Within a certain range,the increasing of parameters can improve the transfection efficiency,but when the parameters were too large (eg.the intensity＞1.6 W/cm2 or 50％ duty cycle),the actual transfection efficiency decreased due to the increased cell death.When the ultrasonic intensity was 1.2 W/cm2,the transfection efficiency and mortality were better than the other subgroups of group A.When the duty cycle was 20％,the transfection efficiency and mortality rate were better than the other subgroups of group B.Continue to increase the exposure time over 90 s was not statistically significant (P ＞0.05) for transfection efficiency and cell mortality.Conclusions The ultrasound-targeted microbubble destruction is an efficient method for gene delivery in vitro.The transfection efficiency vary greatly under different parameters,so optimization parameters is conducive to the promotion of gene transfection.

Objective To compare bolus infusion and replenishment using real low mechanical index contrast enhanced ultrasound in assessing the change of renal cortical perfusion.Methods Using dopamine (i.v.) at the dose of 0.5,2.0,5.0μg · kg- 1 · min- 1 to change renal blood perfusion of 20 rabbits,then during bolus or contant injection of SonoVue,at coded pulse inversion mode,real-time contrast ultrasound was performed,the latter method needed destroying microbubble at a high MI when amplitude reach a steady state,then recording the replenishment,peak intensity(A) and time to peak (PPT) were obtained through raw time-intensity curve,and slope rate of TIC(k) was acquired by curve fitting,standard effective renal plasma flow(ERPF) was measured through 4-aminohippuric acid clearancerate method,meanwhile correlations between ERPF and parameters were analyzed,as well as the paired samples t test for each parameter before and after dopamine administration.Results The ascending branchs of raw TIC of bolus infusion increased sharply and were approximately straight,then descended gradually,while that of replenishment looked like two straightlines with different slopes,then stayed horizontal Both the value of A of two methods were positively correlated with ERPF ( r b =0.85,r re =0.66),and were different at the same ERPF,meanwhile the value of TTP were negatively correlated with ERPF( r b =-0.92,r re =- 0.76),and there were no statistically difference between the two methods.k from Gamma fitting was far from correct,while k from exponential fitting was apparently correlated with ERPF ( r re =0.77 ).Conclusions Both bolus injection and constant injection-replenishment method can assess renal cortical blood perfusion,TIC parameters A and TTP represent regional blood volume fraction and microbubble velocity respectively.Bolus-infusion with real low mechanical index is more precise and available.Comparing with k,TTP is more appropriate to reflect perfusion velocity.

Objective To evaluate the teaching effect of seminar method in clinical teaching of department of urinary surgery.Methods A total of 30 clinical medical students were randomly divided into two equal groups:test group with seminar teaching method and control group with traditional teaching method.Results Were evaluated by scores of theoretic test,operating skill and survey after 3 months.Results Scores of theoretic test and operating skill in test group were significantly higher than those in control group(P ＜0.05).There were significant differences in scores of survey between two groups(P ＜ 0.05).Conclusions Seminar teaching method could effectively improve student learning initiatives and teaching effect,and is worth further exploration.

The aim of this research was to study the immunoprotections of Schistosoma japonicum (S. japonicum) DNA vaccines SjRPS4 and SjRPL7 in mice. Fourty Kunming mice were randomly divided into 4 groups (A, B, C and D), and the pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 plasmid DNA vaccines were prepared for experiment. Mice in group A were intramuscularly injected with 100μL normal saline, whereas mice in group B were injected with 100 (g naked plasmid pcDNA3.0 into the quadriceps. Mice in groups C and D were injected with 100μg/100μL eukaryotic recombinant plasmids pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 into the hind leg muscles respectively. The initial injections were followed by two sets of boosters at 2 weeks intervals. In addition, levels of the specific antibodies were detected 2 weeks after the last immunization and all mice were percutaneously infected with 20( 1) S. japonicum cercariae on abdomen. Fourty-two days after the infection, all mice were killed to detect the worm reduction rate and the egg reduction rate. Significant differences of worm burden reduction rates, LEPG reduction rates, IEPG reduction rates and intrauterine eggs reduction rates were observed in both test group (group C and D), comparing with the control groups (group A and B). Results indicated that the DNA vaccines of pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 could induce strong protective immunity against S. japonicum in mice.

Objective To investigate the effects of transplantated tumor capillary permeability under microbubble enhanced diagnostic ultrasound exposure.Methods Fourty eight rats were randomly divided into four groups after Walker 256 carcinomas were subcutaneousely implanted,named the control group (A),microbubbles group (B),ultrasound group (C),microbubbles and ultrasound group (D).Using Evans blue(EB) as the indicator for tumor capillary permeability,the microbubbles were injected through the vein with diagnostic ultrasound frequency as 1.75 MHz,mechanical index as 1.4.The spillover of EB was estimated under confocal laser microscope,meanwhile the contents of EB in the tumor tissues were calculated according to the standard curve and spectrophotometry.Results EB spillover was observed around the capillaries in the group D,while EB spillover was only observable on capillary in group A,B and C.The EB content was also significantly higher than those in group A,B and C.Conclusions Exposure to both diagnostic ultrasound and microbubbles results in increased capillary permeability of rat tumor tissues,which might become a potential therapeutic method on clinical chemotherapy.

Humans ; Hyoid Bone ; pathology ; Male ; Middle Aged ; Syndrome

Objective To study the relation between the HPV6/18 virus infection and the development of pathological changes of cervix. Methods The number of HPV16/18 DNA copies and the expression rate of HPV16/18 E7 mRNA in the pathological cervix were examined by the quantitative fluorescent PCR combined with pathological diagnosis and immunohistochemistry staining. Results The HPV16 infection rates in chronic cervicitis group were much lower (7.4%) than that in the cervical intraepithelial neoplasia (CIN) groups and the cervical cancer group (69.6% and 72.7%), respectively. Statistical analysis showed that the difference of HPV16 DNA copies was not significant between the chronic cervicitis group and CIN groups. In contrast to the above mentioned result, the number of HPV DNA copies between the CIN groups and the cervical cancer group was significantly different. The HPV16 E7 gene expression rates in CIN Ⅰ, Ⅱ, Ⅲ and cervical cancer groups were 0,37.5%,42.9%,63.6%, respectively. Conclusion Ins more common than that with HPV18. The number of HPV16 DNA copies in cervical cancer tissues is markedly higher than that in CIN Ⅱ, Ⅲ groups. The HPV16 E7 mRNA expression is significantly increased in the cervical cancer, and it is more closely correlated to this pathological changes. The quantitative fluorescent PCR can be used to reflect the activity of HPV, and it is a useful method for the screening examination of HPV and for the early diagnosis and treatment of cervical caner.