Objective To investigate the median effective plasma target concentration of propo-fol (Cp50 )needed for lumbar surgery during inhalation-intravenous balanced anesthesia when BIS is 50.Methods Patients aged 40-56 years,scheduled for vertebral pulp ectomy were included.Anesthe-sia were maintained by TCI of propofol,sevoflurane 0.5 MAC,remifentanyl 0.2 μg·kg-1 ·min-1 , and vecuronium 0.08 mg·kg-1 ·h-1 .The target plasma concentration of propofol was initially set at 1.8 μg/ml,and adjusted by the sequential up-and-down methods,based on BIS index.The Cp50 of propofol and its 95% confidence interval (CI)were calculated.Results Twenty-six patients were en-rolled in this study.The median effective plasma target concentration of propofol was 1.61 μg/ml (95%CI 1.52-1.70)μg/ml.Conclusion The Cp50 of propofol needed for maintaining the BIS index of 50 is 1.61 μg/ml during the anesthesia with 0.5 MAC of sevoflurane combined with remifentanyl in lumbar surgery.

Calbindin 2 ; metabolism ; Heart Neoplasms ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Rhabdomyosarcoma, Embryonal ; metabolism ; pathology ; surgery

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

Human adenovirus type 41 (HAdV-41) is considered to be a "fastidious adenovirus". E1-deleted HAdV-41 cannot be rescued or amplified in 293 cells. To propagate recombinant HAdV-41 in 293 cells, the backbone plasmid pAdbone41 was reconstructed. That is, the E3 coding sequence of HAdV-41 was deleted and replaced with the HAdV-5 E4orf6 gene; and the E1A enhancer of HAdV-5 was inserted upstream of the E4 promoter of HAdV-41. Novel adenoviral plasmid pAd41E4EE-GFP was generated by homologous recombination of the shuttle plasmid pSh41-GFP with the modified backbone plasmid in the Escherichia coli BJ5183 strain. Adenovirus HAdV-41-E4EE-GFP was rescued by transfecting 293 cells with linearized pAd41E4EE-GFP. After seven rounds of propagation, viruses were purified by the CsCl ultracentrifugation method. HAdV-41-E4EE-GFP in 1.0 ml with a particle titer of 8 x 10(10) vp/mL was obtained which had a particle-to-infectious ratio of 50 : 1. The genome of HAdV-41-E4EE-GFP was confirmed by restriction analyses and polymerase chain reaction. These results showed that a novel HAdV-41 vector system was established in which recombinant HAdV-41 could be constructed and packaged in 293 cells.
Adenoviruses, Human ; genetics ; growth & development ; physiology ; Cell Line ; Genetic Vectors ; genetics ; physiology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; metabolism ; Recombination, Genetic ; Transfection ; Virus Assembly

Objective: To investigate the inhibitory effect of Undaria pinnatifida polysaccharides (UPPS) on hepatocellular carcinoma HepG-2 cells and its possible mechanism. Method: The effect of inhibiting proliferation and inducting apoptosis of UPPS were determined by means of MTT and FCM. The expression of Bcl-2 and Bax proteins was immunohis to chemcally evaluated after treatment of UPPS. Results: UPPS inhibited HepG-2 cells growth in vitro , significantly higher than the negative control group (P

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.

To compare the expression of CD antigens on immune cells from umbilical cord blood (UCB) and bone marrow (BM) and analyze its clinical significance, the phenotypes of lymphoid cells and nucleated cells from 38 UCB and 10 BM samples were investigated by flow cytometry with double labeling monoclonal antibodies. The results showed that the immature lymphocytes (CD3(-) CD4(+)) were detected in UCB and higher than those in BM; cytotoxic T lymphocytes (CD3(+) CD16(+) CD56(+)) in UCB were significantly lower than those in BM. NK cells (CD3(-) CD16(+) CD56(+)) in UCB were higher than those in BM. The ratio of CD34(+) cells in nucleated cells of UCB was similar to that of BM, however, both the contents of myeloid (CD34(+) CD13(+) and CD34(+) HLA-DR(+)) and lymphoid (CD34(+) CD19(+)) progenitor cells in UCB were lower than those in BM. It is concluded that the immune cells in UCB possess immaturity, which might lead to mild GVHD after UCB transplantation. It is inferred from the higher ratio of NK cells in UCB, GVL will not decrease after UCB transplantation. The lower contents of myeloid and lymphoid progenitor cells in UCB probably accounted for the slow hematopoiesis and immune reconstitution following UCB transplantation.
Adult ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; immunology ; physiology ; Female ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Infant, Newborn ; Lymphocytes ; immunology

The role of MHC class II transactivator (C II TA) in constitutive or IFN-gamma inducible expression of HLA molecules in human malignant hematological cell lines was investigated. The expression of HLA molecules and C II TA protein was detected by Western blot, immunohistochemistry and flow cytometry. The expression of C II TA gene was determined by RT-PCR. The capability of peripheral blood T cell reaction stimulated by tumor cells was monitored by mixed lymphocyte reaction. It was found that the HLAII-positive tumor cells expressed the C II TA quite well, and the expression of HLA I + II was increased in the tumor cells with constitutive or inducible expression of C II TA after induced by IFN-gamma. The tumor cells which did not express C II TA after induced by IFN-gamma were not response to the expression of HLA II promoted by IFN-gamma. It suggests a correlation between the inability of some malignant hematological cell lines in response to IFN-gamma for HLA expression and the deficiency in the inducible expression of C II TA, indicating C II TA might take part in the regulation of HLA I + II expression in the tumor cells, which might play an important role in tumor immunologic escape.
Animals ; Genes, MHC Class II ; genetics ; HL-60 Cells ; HLA Antigens ; biosynthesis ; genetics ; Humans ; Interferon-gamma ; pharmacology ; K562 Cells ; Leukemia ; metabolism ; pathology ; Nuclear Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; biosynthesis ; genetics ; U937 Cells

Allo-cell transplant rejection is associated with class II major histocompatibility complex (MHC II), while its transactivator (namely C II TA) regulates MHC II molecules expression strictly and exclusively. The aim of this study was to investigate the inhibiting effect of C II TA anti-sense RNA on MHC II expression. The cDNA for anti-sense RNA recognizing the 114-523 sequence of C II TA (arC II TA) was obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid (pcDNA3.1B-arC II TA, pD-arC II TA). Raji cells were transfected stably with pD-arC II TA, classic MHC II antigen (HLA-DR, -DP, -DQ) expression was assayed by flow cytometry (FCM). mRNA abundance of C II TA, invariant chain and classic MHC II were detected by RT-PCR. The results showed that compared with control (sense C II TA), the expression inhibition of HLA-DR, -DP, -DQ on pD-arC II TA positive Raji cell was 65.93%, 54.14%, 68.32% respectively. The mRNA contents of C II TA, invariant chain and classic MHC II also decreased. In conclusion, arC II TA inhibited C II TA and thus the family of MHC II molecules were regulated by it, therefore these results provide a novel approach for the control of graft versus host diseases.
Cell Line, Tumor ; Flow Cytometry ; Graft Rejection ; genetics ; prevention & control ; HLA-DP Antigens ; biosynthesis ; genetics ; HLA-DR Antigens ; biosynthesis ; genetics ; Humans ; Nuclear Proteins ; genetics ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; Transfection

Objective: To evaluate the therapeutic efficacy of acupuncture plus Dang Gui Bu Xue Qu Feng Tang for benign essential blepharospasm (BEB). Methods: A prospective randomized controlled trial was performed. A total of 105 participants were randomized 1:1:1 into an acupuncture group, a herbal medicine group and an acupuncture plus herbal medicine group. Participants in the acupuncture group received manual acupuncture treatment, twice a week. Participants in the herbal medicine group received Dang Gui Bu Xue Qu Feng Tang, oral administration, once a day. Participants in the acupuncture plus herbal medicine group received both treatments. The therapeutic effects of the three groups were evaluated after four weeks of treatment. The primary outcome was the Jankovic rating scale (JRS) score, and the secondary outcome was the blepharospasm disability index (BSDI) score. Results: After four weeks of treatment, the JRS total scores significantly decreased in all three groups versus baseline (P<0.05). A greater reduction in the JRS total score was reported in participants in the acupuncture plus herbal medicine group (P<0.05), but there was no significant difference between the acupuncture group and the herbal medicine group (P>0.05). The acupuncture plus herbal medicine group had a greater decrease in the JRS severity score than the herbal medicine group (P<0.05). The reduction in the JRS frequency score was not significantly different among the three groups (P>0.05). The BSDI scores significantly decreased in all three groups versus baseline (P<0.05), but the reduction in the BSDI score was insignificantly different among the three groups (P>0.05). Conclusion: It is effective in the treatment of BEB either to use acupuncture and Dang Gui Bu Xue Qu Feng Tang alone or in combination. The combination therapy shows a more significant effect than either of the treatment alone.