Objective To explore the possibility of using 131I as a targeted therapy method for malignant glioma by infecting U87 and U251 cells with conditionally replicative adenovirus Ad-Tp-E1a-Gp-NIS.Methods Human telomerase reverse transcriptase (hTERT) promoter and glial fibrillary acidic protein (GFAP) promoter were cloned and their transcriptional activities were detected by luciferase assay.The conditionally replicative adenovirus Ad-Tp-E1 a-Gp-NIS was constructed,purified,and transfected into U87 and U251 glioma cells.For these transfected cells,the selective replication ability was evaluated by plaque forming assay,and protein expression was detected by Western blot assay.125I-iodide uptake and exflux,the clonoy formation of 131I-iodide treated cells were also measured.Results Transcriptions activity of the GFAP and hTERT promoters was 59.75％-62.10％ (F =11.89,P ＜ 0.01) in U87 cells and 37.31％-49.00％ (F =5.87,P ＜ 0.05) in U251 cells.The Ad-Tp-E1a-Gp-NIS could be selectively replicated and the hNIS gene was successfully expressed in the hTERT-positive and GFAP-positive glioma cells which showed two protein bands with relative molecular mass of 120 × 103 and 49 × 103 in Western blot assay.After infection with Ad-Tp-E1a-Gp-NIS,the cell ability of 125I uptake was increased by 78.80 (F =2 914.58,P ＜0.01) and 92.48 (F =2 275.91,P ＜0.01) times in U87 and U251 cells,respectively.The GFAP-negative MRC-5 cells could not take in 125I.The in vitro clonogenic assay indicated that,after 131I treatment,more than 90％ of the transfected cells were killed,while only about 65％ (t =11.73-78.33,P ＜ 0.01) of control cells were killed.Conclusions The Ad-Tp-E1a-Gp-NIS has a good ability in selective replication and the enhancement of antitumor therapy effect by increasing tumor-specific iodide uptake in malignant glioma cells.

Objective To improve the method for determination of geniposide in Fructus Gardeniae.Methods The improved method was compared with the pharmacopoeia method.The content of geniposide was determined by HPLC at 238nm detection wavelength with acetonitrile-water(15:85)as mobile phase .Results The improved method in which 50 % methanol was used as solvent was superior to the pharmacopoeia method in which methanol was adopted as solvent.A higher geniposide content was abtained from the former.Conclusion The improved method is convenient and accurate,and can be used to supply reference evidence for the assay of Fructus Gardeniae in the new edition of pharmacopoeia.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Emergency Service, Hospital ; Female ; Humans ; Male ; Middle Aged ; Singapore ; Skin Diseases ; diagnosis ; Tertiary Care Centers ; Young Adult

BACKGROUND: Excellent Iow-antigenicity xenogeneic biological valve scaffold is the premise of constructing tissue-engineered valve by using which kind of acellular methods.OBJECTIVE: To explore the optimal preparation method of making tissue engineered heart valves by meesuring efficiency of different acellular methods and ability to preserve the matrix.DESIGN, TIME AND SETTING: The prospective randomly controlled study was performed at the Central Laboratory of Taian Central Hospital from January 2007 to June 2008.MATERIALS: Sixteen specimens of porcine aortic valves were randomly divided into control, NaCI-sodium-dodecyl-sulfate (SDS),trypsin and triton-X100 groups.METHODS: Specimens in the control group were left intact. Three test groups were decelluladzed with NaCI, trypsin andTriton-X100 respectively.MAIN OUTCOME MEASURES: The gross structure, optical and electron microscope ultrastructure of the decelluarated porcineheart valve matrix was compared. The expression of vascular endothelial cell major histocompatibility complex (MHC)- Ⅰ antigenwas detected by immunohistochemical method.RESULTS: Treatment with NaCL-SDS achieved only incomplete decellularization. The main components of extracellular matdxwere reserved completely, but the fibrous components became unclear and swelling. Treatment with trypsin removed cellscompletely, but caused serious structural alterations, with the presence of swollen collagen fiber, crude edge, widen and irregularfiber interspace. Treatment with Triton-X100 achieved both complete decelluarization and preservation of the matrix structure.Valves following treatment of NaCI-SDS, trypsin and Triton-X100 had certain immunogenicity. However, the immunogenicity ofvalves following treatment of trypsin and Triton-X100 was significantly lower compared with the treatment of NaCL-SDS.CONCLUSION: The decellularization method by Triton-X100 is effective and complete. The Triton-X100 method does not changematrix structure and has low immunogenicity.

BACKGROUND: Antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70 are the main self-antibodies involved in systemic scleroderma (SSc) and are closely connected with the development of SSc.OBJECTIVE: To probe the value of various self-antibodies and proteins in evaluating the functional prognosis of patients with SSc. DESIGN: Case-control study.SETTING: Clinical Laboratory of Second Hospital Affiliated to Jiangxi Medical College.PARTICIPANTS: Totally 74 patients, 19 males and 55 females aged 12-59years old, were confirmed of SSc at the outpatient and inpatient departments of Second Hospital Affiliated to Jiangxi Medical College between December 1995 and December 2004. There were 46 cases of diffuse cutaneous SSc, 24 cases of localized cutaneous SSc, and 4 cases of overlapping syndrome. Meanwhile40 inpatients (14 males and 26 females aged 19-54years old) who received treatment due to other diseases were recruited from the same hospital.METHODS: The level of antinuclear antibody, anticentromere antibody,and anti-cytoplasm antibody was detected with indirect irnmuneofluorescence assay; antibody against SCL-70 was detected with Western blot; the level of serum immunoglobulin, C-reactive protein (CRP) and rheumatoid factor was examined with velocity dispersion turbidimetry.ticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70;RESULTS: Blood samples collected from the 74 patients with SSc and 40controls were proved eligible and all data entered the final statistical analanti-cytoplasm antibody, and antibody against SCL-70: It was obviously higher in SSc group than in control group [66% (49/74), 53% (39/74), 39%(29/74), 7% (5/74), 0, (x2=57.15, P ＜ 0.01)]. The positive rate of antinuclear antibody in patients with diffuse cutaneous SSc was significantly lower than in patients with localized cutaneous SSc [57% (26/46), 83%(20/24),(x2=5.03, P ＜ 0.05)], but the positive rate of antibody against SCL70 was significantly higher than localized cutaneous SSc [48% (22/46),rheumatoid factor in patients with SSc was markedly higher thanin control group [(16.89±11.94), (11.89±2.05) g/L; (23.06±6.18), (22.44±5.53) IU/mL,t=8.01, 2.46, P ＜ 0.01].CONCLUSION: The positive rate of antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70, as well as the level of immunoglobulin G and rheumatoid factor were obviously increased in patients with SSc, suggesting that these experimental parameters have the value in evaluating the prognosis of SSc.

Objective To understand the pathogenetic role and clinical significance of interleukin 6(IL－ 6 ),tumor necrosis factor(TNF) and soluble interleukin－ 2 receptor(sIL－ 2R) in patients with cerebral infarction(CI) .Methods The IL－ 6,TNF and sIL－ 2R levels were measured in 46 CI patients with ELISA. Results The serum IL－ 6,TNF and sIL－ 2R levels of CI group were much higher than those of controls group .The variation of IL－ 6,TNF and sIL－ 2R levels was closely related to the size of cerebral infarction. The serum IL－ 6,TNF and sIL－ 2R levels of recovery period were much lower than those of acute period in patients with CI .Conclusions The IL－ 6 ,TNF and sIL－ 2R take part in the pathologic process after CI .The measurement of IL－ 6, TNF and sIL－ 2R is useful in determining the size of cerebral infarction.

Objective Using quantitative real-time PCR to establish a rapid specific genetic diagnostic technique for Yersinia pestis.Methods ①Four sets of specific probes and primers were designed,which targeted to chromosome genes of YPO0392,YPO1094,YPO2087 and YPO2090,respectively.②The probes and primers were tested for stability and specificity with 40 strains of Yersinia pestis and 47 strains of non-Yersinia pestis of different sources in Yunnan.③Eight positive DNA in Yulong,Yunnan,were tested with the screened probes and primers.Results ①Two sets probes and primers were selected,they were targeting YPO0392 and YPO1094,respectively.②The results were all positive of the eight positive DNA samples tested.Conclusion Two sets of primers and probes are selected for rapid specific diagnosis of Yersinia pestis.

Objective To study the effects of glycogen synthase kinase-3?(GSK-3?)and free radical on neuron apoptosis of hypoxic-ischemic brain damage(HIBD)in newborn rats.Methods Eighty 7-day-old neonatal rats were randomly divided into 2 groups:normal group and hypoxic-ischemic(HI)group.Rats in HI group were subjected to left common carotid artery ligation,exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for 2.5 h.Rats in 2 groups were killed at 6 hours,24 hours,48 hours,72 hours,5 days after hypoxia respectively.The neuron apoptosis was detected by flow cytometry.The activity of GOD-PX and the contents of SOD and MDA were detected by spectrophotometry,the level of GSK-3? was mensurated by Enzyme-linked immumosorbent assay(ELISA).Results The rates of neuronal apoptosis in HI group were significantly higher than those in normal group at 6,24,48 and 72 hours after hypoxia,respectively(Pa

ObjectiveTo study the assistant therapic effect of the Botulinum toxin A (BTA) on tiptoe after cerebral palsy(CP).MethodsMulti point injection of BAT into the musculus gastrocnemius combined with the rehabilitate therapy was used on 35 children(39 feet)with tiptoe after CP.The therapeutic effect was evaluated after a month.ResultsThe efficiency rate is 25.6%,the slight efficiency rate is 71.8%,the ineffective rate is 2.6%.There is no toxic or side effect was observed.Conclusions The BTA can relieve spasm of musculature and redress abnormality from convulsing of musculature.

Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.