Objective The high quality RNA from the plantlet of Dendrobium candidum was isolated efficiently.It laid a foundation to study the mechanism of molecular expression on its growth and metabolism under environmental stress.Methods The groups stimulated by sound wave and the control groups were established.Total RNA was prepared by a modified phenol-chloroform method.The content was determined by a spectrophotometer.The integrity of RNA was detected by electrophoresis with 1.0% denatured agarose gel.The total RNA obtained was amplified by RT-PCR.The differential cDNA bands were displayed on 6% denatured polyacrylamide gel by electrophoresis and silver staining.Results The ratios of total RNA A_(260)/A_(230) and A_(260)/A_(280) were 3.16 and 1.96,respectively.The yield was 0.27 ?g/mg fresh weight.The bands were clear on agarose gel and the integrity of RNA was good.The differential cDNA bands were screened with molecular weight of 240—600 bp.Conclusion The method of modified phenol-chloroform can be used to isolate RNA from D.candidium with high quality and high yield.

Objective To study the curative effects of surgical treatment of the severelv displaced supracondylar fracture of the humerus in children. Methods A retrospective investigation was conducted on 316 children with severely displaced supracondylar fracture of the humerus who had been treated with operation in our hospital from June 2001 to June 2006.There were 215 boys and 101 girls,with an average age of 7 years old(range,1 to 14).All were treated with open reduction and internal fixation by Kirschner wire,plaster external fixation after operation,and functional training 2 to 4 weeks postoperatively. Results All the 316 cases were followed up for over 6 months to 5 years, with an average of 2 yearS and 10 months. The functional recovery of the postoperative elbow joint was evaluated by Flynn's criteria.Of them,253 cases were rated as excellent,34 as good,25 as satisfactory,and 4 as poor.Cubitus varus deformity was found in 27 cases. Conclusions Although closed reduction and external fixation is the frst choice for the pediatric supracondylar fracture of the humerus, open reduction and Kirschner wire fixation is necessary for the severely displaced supracondylar fracture of the humerus,including those suffering from failed manipulative reduction,open fracture,vascular and neural injuries and limited function of the elbow joint. Anatomic reduction of broken ends,Kirschner wire internal fixation and early functional training are significant in the therapeutic efficacy of operation,and in the prevention or reduction of postoperative complications.

Cone-bine CT(CBCT)images of 2 130 maxillary third molars were divided into 3 groups according to the impaction position. The relationship between the floor of maxillary sinus and the roots of maxillary third molar was analyzed and compared.In low,median and high impaction groups the probability of the root tip of maxillary third molar higher than the floor of maxillary sinus was 4.5%,10.3% and 14.6% respectively.

Objective To study factors that may increase the expression of human ? synuclein gene in prokaryotic cells. Methods proEXNACP vector was transferred into competence cells. Factors such as IPTG levels, initial concentration of cells, type and pH of media, time and temperature of induction were studied in relation to the production of ? synuclein. Results IPTG concentration and pH change does not have a significance impact on ? synuclein production. But the time and temperature of induction, media and initial concentration of cells significantly affected the expression.Conclusion The best condition for the production of ? synuclein includes factors such as LB media with an initial concentration of cells of 0 5\|0 8 A at 37℃ culture with oxygen of 180?rpm/min for 1 2?hours.

OBJECTIVE To compare the effectiveness of two laboratory detections methods of Epstein-Barr virns (EBV). METHODS Immuno-fluorescence technique was applied to detect the serum EBV-specific antibody and EBV-DNA was identified by fluorescence quantitative polymerase chain reaction (FQ-PCR) in 124 children infected with EBV. RESULTS The positive rates of VCA-IgM,VCA-IgG,EA-IgG,EBNA-1-IgG and EB-DNA in various samples of 124 cases were 0,89.5%,23.4%,49.2% and 30.3%,respectively,with 21 cases (16.9%) showing simultaneously positive in EB-DNA and EBNA-1-IgG. CONCLUSIONS The two laboratory detections of EBV share their advantages and disadvantages. FQ-PCR is a method for time-saving,accurate and sensitive detection of EBV-DNA,showing clinical significance in the diagnosis of EBV-associated diseases. It's worthy of clinic application.

Objective To investigate the relationship between the change of HDL level and the severity in patients with sepsis.Methods The levels of HDL were measured in 68 patients with sepsis who were divided into light,middle and heavy group by APACHEⅡscore after final diagnosis of 0,1,2,3,7,14.28 days.Results The levels of HDL in patients with sepsis were lower than those of the normal:the levels of HDL began to decrease at 0 day,got to lowest point after 3 days,and up-regulated gradually after 7 to 28 days.The levels of recovery were the highest in light group,and the lowest in heavy group.Conclusion The levels of HDL in patients with sepsis are low,which is related with the severity of sepsis.

Objective To observe the clinical features of abdominal aortic aneurysm(AAA)in the elderly. Methods Seventy-eight elderly cases with AAA were studied retrospectively,the effect of age and AAA size on the growth of AAA were analyzed. Results Risk factors such as smoking,hypertension,hypercholesteremia and artherosclerosis were found in most patients. Aneurysm in other side was found in 25.6%of the patients.Rupture occurred in patients with a larger in size or rapidly developing AAA.The average AAA diameter inerease was 0.1 4 cm/year and it was similar for each age grade.When the AAA diameter was wider than 5 cm,it developed faster.and the growth velocity increased to 0.46 cm/year. Conclusions Age is not a risk factor that affects the development of AAA.If the AAA diameter is≥5 cm,the AAA develops quickly,then active interventions are recommended.

Objective To construct baculovirus expression system for the human plasminogen kringle 5(hK5) gene,and detect its expression in Spodoptera frugiperda 21(sf21). Methods Baculovirus Bacmid-rhK5 was prepared with bac-to-bac baculovirus expression system,and sf21 cells were infected.The expression of rhK5 in sf21 cells was determined by Western blot.The activity of purified protein was detected by vascular endothelial cell inhibition test. Results The pFastBac HTB-rhK5 was successfully constructed,and sf21 cells were transfected with the constructed vector.The output of rhK5 obtained was 90 ug/L,and the protein possessed the inhibitory activity of endothelial cell proliferation.The median effective dose(ED50) was 4 ug/mL. Conclusion The rhK5 baculovirus expression vector is highly expressed in sf21 cells,which lays a foundation for the the expression of rhK5 protein with biologic activities.

Objective To investigate the protective effect of desflurane preconditioning on human umbilical vein endothelial cells against anoxia-reoxygenation(A/R)injury.Methods The human umbilical vein endothelial cell line(ECV304)was provided by the Liver Cancer Institute,Zhongshan Hospital,Fudan University.ECV304 cells were randomly divided into 5 groups:group Ⅰ normal control;group Ⅱ A/R;group Ⅲ A/R+rhTNF-α;group Ⅳ Des + A/R and group Ⅴ Des + A/R + rhTNF-α.In group Ⅱ-Ⅴ the cells were exposed to 95% N2 + 5% CO2 in an incubator for 30 min followed by 60 min reoxygenation.In group Ⅲ and Ⅴ rhTNF-α(10 ng/ml)10 μl was added to the cells as soon as reoxygenation was started,while in group Ⅳ and Ⅴ the cells were pretreated with 7.2% desflurane for 30 min followed by 10 min washout before A/R.Apoptosis in endothelial cells was detected by fluorescence flow cytometry and TUNEL.Endothelial cells were examined with electron microscope for apoptosis and necrosis.Results The rates of apoptosis in the endothelial cells were significantly higher in A/R group and A/R + rhTNF-α group than in control group.Desflurane preconditioning significantly attenuated apoptosis in the endothelial cells induced by A/R and A/R + rhTNF-α respectively.Electron microscopy showed that there were significantly more necrotic cells in A/R group and A/R + rhTNF-α group.However in the two desflurane preconditioning groups(Ⅳ and Ⅴ)the cells were in a state of duplication and self-repairing.Conclusion Preconditioning with 30 min 7.2% desflurane can attenuate the A/R-induced injury to human umbilical vein endothelial cells.

Objective To identify the cell death type and investigate the potential mechanism of ionizing radiation-induced neural cell death in the neonatal rat cerebral cortex.Methods The neonatal Wistar rats were given a single dose of 2.0 Gy γ-irradiation.The cell death type and characterization in cerebral cortex were identified using DNA electrophoresis,TUNEL and HE staining.The P53-and iNOS-positive cells were analyzed quantitatively using immunohistochemistry.Results The DNA and morphological characterization of death cells indicated that 2.0 Gy γ-irradiation induced apoptosis of the neural cells in neonatal rat cerebral cortex.The apoptosis indices in different cortex regions were significantly increased 4 h after irradiation,and reached the peak value at 12 h post-irradiation.The apoptosis index of neoconex was much higher than that of hippocampus(archicortex)and paleocortex,while paleocortex had lower apoptosis index than hippocampus.The quantitative immunohistoehemistry suggested that the numbers of P53 and iNOS-positive cells were not different between these three cortex regions at the same time-point after irradiation.Conclusion 2.0 Gy γ-rays induced apoptosis of the neural cells in neonatal rat cerebral codex.The response of cells to the damage effects of ionizing radiation was similar in different cortex regions;however,the apoptosis indices were different significantly.These findings imply that the developing phase or type of neural cells may play a pivotal role in the apoptosis process induced by ionizing radiation.