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

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 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 explore the effect of different doses of tirofiban for PCI, myocardial injury and arterial flow conditions.Methods 70 patients undergoing PCI elevation acute myocardial infarction, were randomly and equally divided into the observation group and the control group.The control group of patients before PCI given haplotype character loading dose of tirofiban, the observation group were given a double load before PCI tirofiban.Recording and analyzing two groups of patients cTnI (cardiac troponin I), 90 minST section down percentage circumstances.Results The patients after 6h, 12h, 24hcTnI contents were (2.11 ±0.50,3.50 ±1.64,3.28 ±1.15) ng/mL was significantly lower than the control group (4.09 ± 1.13,9.48 ±2.61,5.79 ±1.26) ng/mL, and the difference was significant(P<0.05); the observation group were CTFC, 90 min fall within ST respectively was significantly better than the control group ( P<0.05 ) .Conclusion Preoperative use of double loading dose of tirofiban can effectively improve the blood flow after PCI,, and reduce the incidence of myocardial injury and postoperative cardiovascular events.

Objective To evaluate the effect on treatment of multiple myeloma complicating femoral neck fracture by surgical intervention combined with drugs.Methods Twelve patients with multiple myeloma complicating femoral neck fracture were treated by cemented total hip arthroplasty femoral head shank.All of patients were received zoledronic acid treatment,at the same time,7 cases of them treated by MPT regimen (melphalan,methylprednisolone and thalidomide),3 by bortezomib combined with dexamethasone,and 1 by autologous peripheral blood stem cell transplantation.Results Twelve patients could tolerate surgery,postoperative pain was significantly reduced.Assessed by using Harris hip function score after 6 months operation,2 cases were excellent,7 cases were good,3 cases were common,excellent and good rate was 75％ (9/12).All patients were followed up for 8 months to 3 years,1 case of local recurrence after 13 months.1-year overall survival rate was 100％,2-year overall survival rate was 83％,and 3-year overall survival rate was 67％.Conclusions Multiple myeloma complicating femoral neck fracture,using bone cement in total hip arthroplasty femoral head shank to clear the local tumor lesions,receive reconstruction capability,rapid postoperative recovery,pain relief,and opportunities for further comprehensive treatments.The bisphosphonate can promote new bone formation and prevent further fractures.The surgical intervention combined with chemotherapy could relieve the symptoms,reduce tumor cell burden,improve quality of life and prolong survival time.

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.

To study the effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on PC-3M cell line, PC-3M cell line was incubated with gradient concentrations of TRAIL for 4--24 h. Annixin-V fluorescence staining and TUNEL method were employed to detect the apoptosis of PC-3M cells. The morphology of apoptotic PC-3M cells was observed by electron microscopy. The relationship between TRAIL concentrations and the percentage of apoptotic cells was evaluated by flow cytometry. The proliferation inhibitory ratio was calculated by using MTT colorimetry. Our results showed that apoptosis of PC-3M cells could be induced by treatment with TRAIL for at most 4 h. The results of flow cytometry and MTT colorimetry demonstrated a time- and concentration-dependent relationship between cell apoptosis rate and TRAIL concentration. It is concluded that apoptosis of PC-3M cells can be induced by TRAIL. Because of the selective killing effect of TRAIL on tumor cells, it may become a potential alternative for the treatment of advanced prostate cancer.
Apoptosis/*drug effects ; Cell Line, Tumor ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/*pathology ; TNF-Related Apoptosis-Inducing Ligand/*pharmacology

Background Diabetes is one of the risk factors that leads to corneal neuropathy.Silent signal regulatory factor 1 (Sirt1) plays an important role in glucose metabolism,lipid metabolism,regulation of insulin secretion and is closely related to the nervous system disease.The relationship between Sirt1 and diabetic corneal neuropathy is not fully understood.Objective This study was to detect the expression of Sirtl in cornea and trigeminal ganglion with type 1 diabetes model mice and explore the association of Sirt1 expression with diabetic corneal neuropathy.Methods Eight C57BL/6-Ins2Akita/J male mice and eight wild-type C57BL/6 male mice in the same litter were selected as type 1 diabetes model group and control group,respectively.The mice of two groups were sacrificed in overdose anesthesia method at 12-month old.Histological examination of cornea and trigeminal ganglion was performed using hematoxylin and eosin staining.Expression and localization of Sift1 protein in cornea and trigeminal ganglion were detected using immunohistochemistry.Western blot assay and fluorescine quantitative PCR were respectively used to quantitatively analyze the expression of Sirt1 protein and Sirt1 mRNA.Results Trigeminal ganglion cells were uneven in size and shape with the loosened cellular arrangement and disorder neurofibrosis alignment,and the corneal epithelial cells were less in the C57BL/6-Ins2Akita/J mice,but the trigeminal ganglion cells and corneal epithelial cells were normal in wild-type C57BL/6 mice.Immunochemisty exhibited that Sirtl protein was expressed mainly in corneal epithelium and the expression of Sirtl protein was stronger in the C57BL/6 mice than that in C57BL/6-Ins2Akita/J mice.Fluorescine quantitative PCR assay showed that the gray scale value of Sirt1 mRNA in cornea in C57BL/6-Ins2Akita/J mice was lower than that of the wild-type C57BL/6 mice(0.56±0.03 vs.0.98±0.13) with significant difference (t =5.853,P =0.010).Western blot showed that the expression of Sirt1 protein in cornea was lower in C57BL/6-Ins2Akita/J mice than that of the wild-type C57BL/6 mice(0.78±0.017 vs.1.300±0.012) with significant difference(t =33.140,P =0.001).However,no significant differences were seen in the gray scale value of Sirt1 mRNA(2.45±0.18 vs.2.51±0.22) (t=0.587,P=0.599) and protein level(1.100±0.015 vs.1.110±0.017) (t =0.430,P=0.709) in trigeminal ganglion tissues between C57BL/6-Ins2Akita/J mice and wide-type C57BL/6 mice.Conclusions The corneal nerve and structure is abnormal in 12-month-old C57BL/6-Ins2Akita/J mouse.Sirt1 is involved in the pathogenesis of diabetic keratoneuropathy,suggesting that it may be a potential target.