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

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.

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 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.

OBJECTIVE: To study the efficacy of treatment with microsurgery in combination with local injection of type A botulinum toxin for vocal process granuloma. METHOD: 28 patients with vocal process granuloma received endotracheal intubation under general anesthesia. The lesion was removed with micro-scissor and CO2 laster under a self-retaining laryngoscope and microscope. The incision and mucous membrane surrounding the wound was closed with 8-0 absorbable suture. 4-point injection of botulinum toxin type A 8-15 u was then performed along the thyroarytenoid muscle and arytenoid muscle of the same side. Postoperative medication was administered based on disease causes. RESULT: All patients experienced vocal cord dyskinesia of the injected side 2-3 days after surgery. At 1 month after the surgery, wound healing was good in all the 28 patients, and the vocal cord movement was limited at the injected side. At 3 months, movement of the bilateral vocal cords was normal, and the vocal cord process mucosa was smooth. Patients were followed up for more than a year, and only one patient had recurrence in 2 months after surgery. The cure rate was 96. 4%. CONCLUSION Combination of laryngeal microsurgery and type A botulinum toxin local injection can shorten the treatment course of vocal process granuloma.
Anesthesia, General ; Botulinum Toxins ; administration & dosage ; Granuloma ; drug therapy ; surgery ; Humans ; Injections ; Intubation, Intratracheal ; Laryngeal Mucosa ; Laryngeal Muscles ; Laryngeal Neoplasms ; drug therapy ; surgery ; Laryngoscopes ; Larynx ; Microsurgery ; Postoperative Period ; Recurrence ; Vocal Cords ; Wound Healing

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 quantify the white matter hyperintensity (WMH) with serial MRI in elderly people with cerebrovascular disease, and to evaluate the risk factors that may have impact on their progression.Methods One hundred and thirty-eight patients with cerebrovascular disease underwent twice MRI scans with a 1.5T MR scanner at least one year apart (mean = 13.8 months). The clinical data of all patients, including age, gender, systolic blood pressure, blood glucose level, serum lipid level, alcohol consumption and smoking were recorded at baseline, as well as the historical informations concerning hypertension, diabetes mellitus and hypercholesterolemia. Besides, the overall severity of cognitive impairment and neural deficit of patients were rated by Mini-Mental State Examination (MMSE) and the National Institute of Health Stroke Scale (NIH). MRI measures included volume of gray matter infarction, volume of white matter infarction, and baseline volume of WMH. The volumetric changes of WMH between the twice scans were assessed using a semi-automated software. The influence of risk factors on changes of WMH volume was analyzed. Correlation coefficients were calculated between clinical scales and the change of WMH volume. Results The Baseline WMH volume was(13155?18782) mm3, and the volumetric change of WMH was(7687?9079) mm3. Multiple regression analyses revealed that the occurrence of infarction in cortex and in white matter was significantly associated with the volumetric change of WMH, as well as the baseline WMH volume. The volumetric changes of WMH were related to MMSE and NIH score (r=-0.266,P=0.002; r=0.257,P=0.002). The total infarcted volumes were associated with the volumetric change of WMH (r=0.416,P

AIM:To establish the quality standard of Compound Shouwu Granule (Radix Polygoni multiflori,Radix et Rhizoma Thalictri Baicalensis,Herba Rabdosiae Rubescentis,etc.) METHODS:TLC was performed to identify Radix Polygoni Multiflori,Radix et Rhizoma Thalictri Baicalensis,Herba Rabdosiae Rubescentis.HPLC was used to determine 2,3,5,4'-tetrahydroxystilbene-2-0-?-D-glucoside. RESULTS:The study on the quality control showed that the characteristic of identification by TLC was distinct and highly specific.The quantitative evaluation had the linear range of 0.04-0.72 ?g.The average recovery was 99.99% and RSD was 1.3%. CONCLUSION: The method for identification and quantitation was simple,accurate,realizable and reproducible.It can be used effectively for the quality control of Compound Shouwu Granule.