The effect of endothelin (ET-1) on the function of the isolated perfused working guinea pig has been examined. ET- 1 wasshowed to exert a transient positive inotropic effect on the isolated heart, followed by a marked inhibition of cardiac function. Low dose of ET- 1(30,90 pmol?L-1) produced reversible inhibition. The effect of high dose of ET- 1 was irreversible, accompanied with increasing LDH significantly, indomethacin markedly augmented the vasoconstrictor and cardiac ischemia effects of ET-1. Our results suggest that the apparent inhibition of cardiac function on the isolatedperfused guinea pig heart after administration of ET-1 may be due to the effect of the myocardial ischaemia caused by progressive development of coronary vasoconstriction. In our experimental model, PGI2 has partially antagonizing effect on the vasoconstrictor action of ET-1.

Objective To establish a model of injury to primarily cultured spinal cord neurons,mimicking the neuronal injury after complete transactional spinal cord trauma,for the sake of exploring changes in expression of an immediately-early gene,c-jun,in central nervous system injury. Methods Spinal cords were removed form fetal Wistar rats at the 14th gestation day and the neurons were cultured for 10 to 12 days.Then,mechanical injury was applied to the neurons by making regular scores on the culture disk under direct vision with the aid of a self-made standard template.Morphology of the injured neurons and changes in expression of c-Jun protein were observed before and at different intervals after injury. Results c-Jun expression was noted in neuronal nuclei 10 min after injury and its peak appeared at 2 hrs.Besides,the density of positive neurons bore evidently an inverse proportion with their distance from the scores.Conclusion\ Positive expression in injury neurons show that c-jun gene enters the nucleoli of injured neurons and takes the role of “the third messenger” at early time after neuronal injury.

Objective To promote rational drug use and assure the drug use in an effective and safe way by developing the monitoring role of clinical pharmacists in the adverse drug reaction .Methods The common reasons for adverse drug reaction were an-alyzed.The clinical pharmacists took advantage of their professional knowledge to deal with adverse drug reactions and reinforce the monitoring of the drug treatment.Results The clinical drug safety was ensured by reducing the adverse drug reaction incidence rate. Conclusion Pharmacy information exchange should be enhanced and the professional knowledge of clinical medical staff should be broadened to promote clinical rational drug use.

Objective To investigate the apoptosis rate of neurons in vitro among the primary culture cells isolated from rat spinal cords before and after injury. Methods The spinal cord neurons of Wistar rats at 14-day gestation were isolated and cultured. The neuronal processes were then injured by cutting. At different time points after injury, TUNEL method was employed to detect the apoptotic neurons. Results Before injury, there was almost no apoptotic neuron. However, a large amount of apoptotic neurons were observed after the injury. The highest incidence of apoptosis appeared on the first day, and then it gradually reduced in the following days, but increased in amount again on the seventh day. Conclusion The results of the present experiment reveal the regularity of apoptosis of neurons before and after injury, and it provides a platform for further research regarding therapeutic intervention in the treatment of spinal cord injury.

Objective To explore the protective effect of HGF on primary cultured spinal neurons in vitro.Methods Cultured neurons were transfected with Ad-GFP or Ad-HGF in different multiplicity of infection(MOI),then flow cytometer and PI-Hoechst double stains were used to assay transfer rate or to determine the status of cells respectively.ELISA was used to detect the expression of HGF in Ad-HGF transfected neurons,and the activity of neurons was judged by neutral red stain,MTT and NSE-ELISA methods.Results After transfection with Ad-GFP in MOI of 50,the transfer rate was high as detected by flow cytometer,and the status of cells was well as judged by PI-Hoechst double stain.The results of ELISA showed that HGF was expressed in medium.After transfection with Ad-HGF,the activity of neurons was better than neurons without treatment(P

BACKGROUND: Hepatocyte growth factor (HGF) promotes neurite outgrowth from neocortical explants, and supports neuronal survival under serum-free condition. Thus, HGF can mediate neurotrophic function as a novel neurotrophic factor.OBJECTIVE: To establish an in vitro injury model with a semi-solid culture system for the purpose of improving the evaluation of neurite regeneration of transected spinal cord neurons from rat embryo, and investigate the effect of HGF on neurite regeneration.DESIGN: Randomized controlled study.SETTING: Hematology Laboratory of Radian Medical Institute of Academy of Military Medical Sciences of PLA.MATERIALS: The experiment was carried out at the Hematology Laboratory of Radian Medical Institute of Academy of Military Medical Sciences of PLA from August 2004 to May 2005. Wistar fetal rats of 14-16 days old were provided by Animal Center of Academy of Military Medical Sciences of PLA. Tail collagen was extracted from adult male Wistar rats with body mass of (250±50) g.METHODS: ① Rat tail type Ⅰ collagen substrate was prepared and spread on a culture dish, cut into about 0.5-1.0 mm3 slices, then spinal cord slices of 15-day-old fetal Wistar rat were explanted on the primary culture. Five days later, the outgrowing processes were severed, then a block of collagen, with the surface area of 2 mm2 and 200 μm away from the slice, was removed and the vacancy was replaced with a fresh collagen block of 2 μL after aspirating the medium. The fresh collagen block could be solidified and then fresh liquid medium was added as the secondary culture. The regeneration of neurite was observed by microscopy at 0, 1, 6,12 and 24 hours after severing. ② The medium was changed with 0.5% N3-conditioned medium. 10 μg/L HGF was added in the experimental group, and 0.5% N3-conditioned medium was added in the control group.The status of regeneration was evaluated by the average value of 3 longest regenerative neurites for each slice. There were 12 slices in each group.The status of neurite regeneration was calculated and was evaluated 24 hours later.MAIN OUTCOME MEASURES: ① neurite regeneration in situ; ②comparisons of neurite regeneration between control group and experimental group.RESULTS: ① Neurite regeneration in situ: The neurites disintegrated near the severing line immediately following the transection injury. This process persisted about 1-2 hours and the distance away from the severing line was about 20 μm. Then the proximal end of neurites would swell and thicken. At this time neurites stopped collapsing and neurite regeneration began. Their regenerating rate would quicken at 12 hours after severing. Regenerating neurites were more branching and curlier as compared with original neurites. ② Comparisons of neurite regeneration between control group and experimental group: The average length of regenerative neurites was more in the experimental group than that in the control group [(375±96) μm, (200±75) μm, P ＜ 0.05].CONCLUSION: ① We establish a simple, economic model to evaluate neurite regeneration. ② By this model, we prove that HGF can promote neurite regeneration.

Objective To evaluate the STR profiling availability of formalin-fixed tissues and the detectable time limit of intact STR profile.Methods The different human tissues were fixed with 10-fold diluted commercial 40%formalin fixative for different duration under 15～20℃,and then DNA was extracted using the method of QIAamp~(R)DNA and IQ~(TM) DNA System.The extracted DNA was quantified with QuantifilerTM kit and amplified by both AmpFSTR identifiler kit and AmpFSTR MiniFiler kit.The STR profile was analyzed by GeneMapper ID v3.2 on 3100-Avant.Resuls The STR profiling availability of formalin-fixed tissues was relevant to the formalin fixing duration mainly.as well as the type of tissues and the template concentration and protocol of DNA extracting.The optimal ranges of template concentration is 1～3ng/μL and the QIAamp extracting method was preferable.There are differences in the degradation rate between various types of tissues in the unbuffered formalin fixative,and the lung tissue showed the slowest rate and liver and intestine tissues the fastest.Intact miniSTR profile of all the tissues detected could be obtained within 15 days duration of formalin fixing while intact STR profile could be obtained within 4 days.Conclusion The major factor that impact the availability of STR profiling of formalin-fixed tissues is the fixing duration in unbuffered formalin,as well as the type of tissues,method of extraction,concentration of PCR template and the kinds of STR loci.

BACKGROUND:During fracture healing, in addition to the need for appropriate biomechanical environment, the role of cytokines is also increasingly attracted attention.
OBJECTIVE:To study the effect of nerve growth factor and alpha-lipoic acid on fracture healing in rat models of femoral fracture.
METHODS:Sprague-Dawley rat models of femoral fracture were established. Seventy-two rats were randomly divided into three groups. In the control group, rats were intramuscularly injected with physiological saline. In the nerve growth factor group, rats were intramuscularly injected with nerve growth factor 200 ng/kg, once a day. In the combined therapy group, rats were intramuscularly injected with nerve growth factor 200 ng/kg and oral y taken alpha-lipoic acid 25 mg/kg, once a day. At 1, 2 and 3 weeks after administration, bony cal us volume was measured. Enzyme linked immunosorbent assay was used to measure serum bone morphogenetic protein-2 levels. Western blot assay was utilized to detect bone morphogenetic protein-2 protein expression at the broken end of fracture. Semi-quantitative RT-PCR was applied to examine vascular endothelial growth factor mRNA expression.
RESULTS AND CONCLUSION:(1) At 1 week after administration, no significant difference in bony cal us volume was detected among the three groups. Serum bone morphogenetic protein-2 level, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA expression were significantly higher in the nerve growth factor group and combined therapy group compared with the control group (P<0.05), but no significant difference was found between the two groups. (2) At 2 weeks after administration, the amount of cal us, serum bone morphogenetic protein-2 levels, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA levels were significantly higher in the nerve growth factor group and combined therapy group compared with the control group (P<0.05). Above expression levels were higher in the combined therapy group than in the nerve growth factor group (P<0.05). (3) At 3 weeks after administration, serum bone morphogenetic protein-2 levels, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA levels were significantly decreased in the nerve growth factor group. However, above expression levels were stil high in the combined therapy group, and significantly higher than in the nerve growth factor group (P<0.05). (4) These results indicate that nerve growth factor combined with alpha-lipoic acid had better effects on the fracture healing compared with the nerve growth factor alone.

To study the mechanisms whereby cisplatin suppresses survival of human esophageal squamous cell carcinoma cells.The cytotoxicity of cisplatin in cisplatin-resistant cell line EC109 /CDDP and its parental cell line EC109 was measured by cell viability assay.Western blotting was used to investigate the protein expression of to-tal p53 and phosphorylated p53 at Ser15.Colony formation assay was employed to evaluate the ability of cells to recover from treatments and form colonies.The results indicated that EC109 /CDDP cells were more resistant to cisplatin-induced cytotoxicity than EC109 cells,with the IC50 values of (20.4 ±4.4)μmol /L and (5.7 ±0.1 )μmol /L,respectively.Although cisplatin did not alter the total protein level of p53,it obviously increased the phosphorylation of p53 at Ser15.Cisplatin inhibited survival of both EC109 /CDDP and EC109.Notably,inhibition of p53 by Pifithrin-αsignificantly promoted recovery of cisplatin-treated EC109 and EC109 /CDDP cells to differ-ent degrees.In this respect,p53 protein was found to be activated in response to cisplatin treatment in both EC109 /CDDP and EC109,which may contribute to the cytotoxic effect of cisplatin.

To explore the clinical manifestation of lymphomatoid granulomatosis (LG) and the early diagnosis. Retrospective analyses of the clinical features of LG and pathology features by open lung bio-psies were conducted. The patient was a 42-year-old female with irregular fever,and her chest X-ray and computed tomography showed nodules with cavity and pleural effusion. Open lung biopsy proved LG. LG is seldom seen in clinic. Open lung biopsy is very important for pathology diagnosis. Early diagnosis and treatment are the key to improving survival in these patients. The therapeutic effect is good.