Messenger RNA was isolated directly from lung cancer cell line A549 using magnetic particles. First strand synthesis from mRNA was driven by M-MLV(Moloney Murine Leukemia Virus) reverse transcriptase and random hexam-eric primer, followed by second strand synthesis using RNase H and DNA polymerase I After treatment with T4 DNA polymerase to flush the ends, the double-stranded cDNA was cloned into the plasmid expression vector digested with EcoRI and followed by removing cohesive end. The number of independent clones of the resulting cDNA library was about 9.0 x 105. The estimated percentage of colonies with inserts was about 85 % . The insert size ranges from 0.5 kb ~ 4 kb. The CPP32 gene coding for death protease was obtained by PCR with the cDNA library from lung cancer cells as a template for the first time. Construction of the cDNA library laid a foundation for screening other genes regulating death of lung cancer cells.

Objective A retrospective study was conducted to evaluate the clinical significance of Video EEG monitoring in the diagnosis of epilepsy and other paroxysmal events Methods Video EEG monitoring under the state of awaking and sleeping and evoked tests were performed continuously in 216 patients with epilepsy and other paroxysmal events Results The characteristic events were captured in 130(60%) of the 216 patients And out of the 130 patients, 53 had clinical events accompanying epileptiform discharge 73 patients had no epileptiform discharge in both ictal and interictal period Seizure types were defined in 80%of 80 patients with epileptiform discharge, and classification was different from the original 42% of the 64 patients Conclusions Video EEG can record much more epileptiform discharge than routine EEG It should be an effective method in both diagnosis and classification of epilepsy

The quality and function of the donor liver is one of the main factors which influence the success and prognosis of liver transplantation.At present,the major source of donor liver for transplantation is Donor of Brain Death (DBD) all over the world,which has unstable circulation.When the brain death occurs,a series of serious physiological function changes will be induced within a few minutes and affect the hemodynamics and homeostasis of the body,which will greatly influence the liver quality and function,and consequently the success and prognosis of liver transplantation,finally leading to the loss of potential donor organs.Therefore,it is necessary to identify the physiological function changes induced by the process of brain death and its injury to liver and take immediate proper protective measures,which can effectively reduce the organ injury,improve liver function and enhance the organ utilization and liver transplantation success.In this paper,the changes and maintenance measures of liver physiological function in DBD will be reviewed.

Objective To investigate the effect of tourniquet compression on axonal transport in sciatic nerve of rats.Methods Twenty-four 12-week old male SD rats weighing 250-300 g were randomly divided into 4groups according to the duration of tourniquet compression(n=6 each):1,2,4 and 12 h.The tourniquet was applied to the middle 1/3 of thigh.In each animal whether the left or right thigh was compressed was determined by a flip of coin.The tourniquet was released for 10 min after every hour of compression.A 3-cm segment of sciatic nerve was removed at the end of tourniquet compression(1.5 cm proximal and 1.5 cm distal to the site of compression).Immuno-histochemistry was used to measure the expression of insulin-like growth factor-1(IGF-1)in the sciatic nerve.The ratio of average optic density of the compressed sciatic nerve to that of control was used to estimate the degree of IGF-1 accumulation.The regression equation of the interaction between the duration of compression and accumulation of IGF-1 was analyzed.Results There was significant accumulation of IGF-1 in the sciatic nerve proximal to the compressed site.The accumulation increased with the duration of compression.There was no significant accumulation of IGF-1 in the sciatic nerve distal to the compressed site.The regression equation of the interaction between the duration of compression(X)and accumulation of IGF-1(Y)was Y=0.422X+0.887.Conclusion Tourniquet compression of sciatic nerve can inhibit axonal transport.The accumulation increases with the duration of compression.

Adolescent ; Child, Preschool ; Eosinophilia ; complications ; Female ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

OBJECTIVE:To study the relation between the level of bacterial endotoxin and the severeness of biliary tract infection METHDOS:Chromogenic technique was adopted to detect the content of bacterial endotoxin in human bile RESULTS:The level of bacterial endotoxin in human bile raised along with severeness of biliary tract infection CONCLUSION:Detection of bacterial endotoxin in bile using chromogenic technique is of help to the clinical judgement of the severeness of biliary tract infection

BACKGROUND: Looking for effective measures to ensure the survival of the implanted stem cells against ischemia-induced hypoxia becomes the major concern in the research of cell transplantation therapy for cerebral infarction.OBJECTIVE: To study the effects of human Persephin gene transfer on hypoxia-induced apoptosis of neural stem cells.DESTGN: A randomized controlled basic study on cells.SETTTNG: Department of Neurology, the Second Affiliated Hospital of Sun Yat-sen University.MATERTALS: This study was completed in the Lin Baixing Laboratory Center of the Second Affiliated Hospital of Sun Yat-sen University from July to December in 2006. Recombinant adenovirus pAdCMV persephin was constructed in our lab. C17.2 neural stem cells were kindly provided by Prof. Snyder, Harvard Medical University, USA. Trypsin and DMEM/F12 were purchased from Gibco Company (USA), fetal bovine serum (FBS) from Sijiqing Biological Engineering Materials Co. Ltd (Hangzhou, China); Poly-lysine from Sigma Company (USA), TUNEL assay kit and FuGENE kit from Roche Molecular Biochemicals Company (Swiss), and S-P immunohistochemical detection kit and DAB reaction kit from Mycine Biological Engineering Company (Fujian). Rat anti-human monoclonal Nestin antibody and rabbit anti-human polyclonal persephin antibody were manufactured by Santa Cruz Company (USA), and persephin anti-senseoligodeoxynucleotide (ODN) was synthesized by Shanghai Biological Engineering Company.METHODS: ① Interventions: C17.2 neural stem cells cultured in vitro were infected by recombinant adenovirus containing persephin gene, and they were divided into four groups: blank control group (Group A, in which the C17.2 neural stem cells were not treated with hypoxia), hypoxic group [Group B, in which the cells were cultured at 37 ℃ in anaerobic incubation containing N2 (0.95 in volume fraction) and CO2 (0.05 in volume fraction)], hypoxia + pAdCMV persephin infection group [Group C, where the cells were cultured under the conditions as in group B after pAdCMV persephin infection for 48 hours], and hypoxia + pAdCMV persephin infection + anti-sense persephin ODN group (Group D, where the cells were infected by pAdCMV persephin and anti-sense persephin ODN. ② Evaluation: The expression of Persephin protein was analyzed using Western blotting; Apoptotic index was detected with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) assay; The changes of apoptotic rate was determined with flow cytometry.MAIN OUTCOME MEASURES: Expression of Persephin protein; Apoptotic index; Apoptotic rate.RESULTS: ① Expression of Persephin protein: A specific band (relative molecular mass of 24 000) was detected by Western blotting in pAdCMV persephin infected cells, suggesting the successful expression of persephin gene.Interestingly, the cells infected with both pAdCMV persephin and anti-sense persephin ODN also showed the specific band of about 24 000, but with much less density, indicating that anti-sense persephin ODN could effectively inhibit the expression of pAdCMV persephin. However, this band was not presented in the blank control groups. ② Apoptotic index:The apoptotic index in group C was significantly lower than those in groups B and D (P＜0.01), but still higher than that of group A (P＜0.01), suggesting that persephin gene transfer could attenuate apoptosis to some extent. ③ Apoptotic rate: The apoptotic rate in groups B and D were obviously higher than that in group A (P ＜ 0.01), and it was lower in group C than in groups B and D (P＜0.01).CONCLUSION:Recombinant adenovirus can efficiently mediate Persephin gene transfer into C17.2 neural stem cells,resulting in high expression of the exogenous Persephin in vitro, which effectively reduces C17.2 neural stem cell apoptosis induced by hypoxia.

In order to survey antibiotic resistance of clinical isolates carbapenem-resistant A cinetobacter baumannii in Meizhou and to investigate resistance mechanism and molecular epidemiological characteristics ,a total of 210 non-duplicated clinical isolates of carbapenem-resistant Acinetobacter baumannii from January 2012 to December 2012 were collected .The K-B disk diffusion method was applied for the drug-susceptibility test ,a modified Hodge test was used for the screening of carbapen-emase ,PCR was used to amplify carbapenemase genes (including IMP ,VIM ,OXA-23 ,OXA-24 ,OXA-51 and OXA-58) ,and the positive products were sequenced .Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) was used for DNA typing and test of homology .Our results on the percentage of strains resistant for antibiotics tested were higher than 60% except for polymyxin B was 0 .48% .There were 163 positive strains by the modified Hodge test ,accounting for 77 .62% .OXA-51 gene was identified in 198 strains (94 .29% ) ,OXA-23 in 165 strains (78 .57% ) ,and VIM in 9 strains (4 .29% ) ,OXA-24 ,OXA-58 and IMP gene was not identified by PCR amplification .Seven genomic types were included in the 210 carbapenem-resistant Acinetobacter baumannii .The major prevalence types were Type A (97 strains) ,Type B (44 strains) and Type H (25 strains) . In conclusion ,multiple drug resistance of clinically isolated carbapenem-resistant A cinetobacter baumannii is a serious problem in Meizhou .Production of OXA-51 ,OXA-23 and IMP carbapenemases is an important mechanism of resistance to carbapenem antibiotics ,and there is prevalence of the same clones in these carbapenem-resistant strains .

Objective To analyze the detection rate and drug resistance tendency of Acinetobacter (A .) baumannii in Meizhou area during 2008-2012 in order to provide the guidance for clinicians′medication .Methods The detection rate and drug resistant rate of A .baumanii in the clinical specimens submitted from 5 hospitals in Meizhou during 2008 -2012 were retrospectively ana-lyzed .The antimicrobial susceptibility test was performed by the disk diffusion method .The WHONET 5 .4 and SPSS18 .0 soft-wares were adopted to analyze the data .Results The detection rates of A .baumanii in these five years were 12 .91% ,15 .40% , 11 .94% ,13 .59% and 14 .00% respectively .In the sources of strains ,sputum had the highest distribution rate of 68 .99% (1 713/2 483) ,in the distribution of departments ,ICU had the highest distribution rate of 33 .91% (842/2 483) .The resistance rate of A . baumannii to cefoperazone/shubatam ,meropenem and imipenem were below 30% in the five years ,but showed the upward tenden-cy .The 5-year total drug resistance rates of A .baumanii to 18 kinds of antibacterial drugs were statistically different between ICU and non-ICU department (P<0 .05) ,the drug resistant rate of isolates from ICU was higher than that from the non-ICU depart-ments .The isolation rate of multi-drug resistant strains of A .baumanii was fluctuated in about 50% during these five years except the lower isolation rate in 2008 ,the isolation rate of pan-drug resistant A .baumanii and carbapenem resistance A .baumanii showed the upward tendency .Conclusion The drug resistance rate of A .baumanii is gradually increased .The drug resistance monitoring of A .baumanii in ICU should be strengthened .Antimicrobial agents should be reasonably used for maximizing to retard the emergence of drug resistant strains .

Objective: To investigate the inhibitory effect of soluble KDR(sKDR) and its antibody on endotheliocyte(EC) proliferation.Methods: The binding of sKDR with VEGF was detected by ELISA. The KDR262 antiserum was prepared from rabbit and the specificity was defined by Western blot. The inhibition of EC proliferation was analyzed by 3H-TdR up-take, MTT and cell counter. Results: The sKDR could bind to VEGF165 specially, the ability was enhanced by heparin. The specific binding of KDR262 antiserum to KDR was also detected. The inhibitory rate of sKDR(10,2,0 4?g/ml) on EC proliferation was 56%,44%,32% respectively. The inhibitory rate of KDR262 antiserum (1∶50,1∶200,1∶800) was 70%, 56%,43% respectively. The cell counter also showed that EC proliferation was inhibited significantly in sKDR group and KDR262 antibody group, vs VEGF165 group, GST group and PBS group. The inhibitory effect was characterized by concentration-dependent and was higher in KDR262 antibody group than that in sKDR group. Conclusion: sKDR expressed by E.coli and its antibody had significant inhibitory influence on EC proliferation.