AIM: To construct the eukaryotic expression vector CD80-IgG by fusing the cDNA encoding extracellular portion of murine CD80 to the 5'-terminus of cDNA encoding Fc fragment of murine immunoglobulin G1 and to express the fusion protein in Chinese hamster ovary (CHO) cells. METHODS: The two cDNAs was amplified by PCR respectively from plasmid pcDNA/B7 containing the full-length cDNA of murine CD80 from murine spleen cells, and cloned to the eukaryotic expression vector pcDNA3.0 by directional cloning. The resultant recombinant plasmid pcDNA/CD80-IgG was transfected into CHO cells with liposome transfection reagent. The stably expressing cells were obtained by G418 screening. Western blot, Dot ELISA, and flow cytometry were used to detect the expression of the fusion protein and its immunological activity. RESULTS: DNA sequencing verified the correction of the construction of recombinant plasmid pcDNA/CD80-IgG. The expressed fusion protein was detected in the supernatant of transfected CHO cells and the molecular weight of the protein was similar to what we expected. Its immunological activity was also established. CONCLUSION: The recombinant plasmid pcDNA/CD80-IgG was successfully constructed and it expressed the fusion protein CD80-IgG.

China ; epidemiology ; Humans ; Occupational Diseases ; epidemiology ; Occupational Health ; Rural Population ; statistics & numerical data

Adult ; China ; Follow-Up Studies ; Humans ; Middle Aged ; Mining ; Rural Population ; Silicosis ; mortality

Adult ; Bronchoalveolar Lavage ; adverse effects ; Female ; Humans ; Intraoperative Complications ; Male ; Middle Aged ; Pneumoconiosis ; therapy ; Retrospective Studies ; Young Adult

Adult ; China ; Humans ; Middle Aged ; Occupational Health Services ; Pneumoconiosis ; prevention & control ; Universal Precautions

Objectives To analyze the relationship between phosphorylation of serine-3 of cofilin1 and Taxol resistance of ovarian cancer and to evaluate the prognostic value of cofilin1 phosphorylation in estimating ovarian cancer survival using clinical samples.Methods Wild type (WT) plasmids with over-expression of wild cofilin1,S3L plasmids with over-expression of cofilin1 and inhibited phosphorylation at serine-3,and siRNA-C plasmids with down-regulation of cofilin1 were constructed using molecular biology techniques.After the SKOV3 and SK-TR30 cell-lines were transfected with these plasmids,changes in biological characteristics and drug resistance after instant transfection were compared.Fifty-one elderly female patients with microscopically confirmed ovarian cancer,who had received chemotherapy with Taxol and cis-platinum (TC)after surgery,including 30 chemo-sensitive and 21 chemo-resistant cases,were recruited in this study.Immunohistochemical methods were used to detect the expression of cofilinl and phosphorylated cofilin in tissue sections from the two groups.Progression free survival(PFS)was analyzed.Results Compared with the control group,the proportion of cells increased at the G0/G1 phase(P=0.034)and decreased at the S phase (P=0.031),and the apoptosis rate decreased(P=0.020),in SKOV3 cells with high expression of cofilin1.However,these characteristics disappeared when the wild type was replaced with the inactive mutant S3L.When cofilin1 expression was down-regulated in SK-TR30 cells,the proportion of cells at the G0/G1 phase decreased(P=0.020).The expression of cofilin1 was detected in 56.7 ％ (17/30)and 57.1％ (12/21),respectively,sections of ovarian tumor tissues of the chemo-sensitive and chemoresistant groups,and there was no statistic difference (x2=0.332,P =0.943)between the two groups.The expression of phosphorylated cofilin was much higher in the chemo-resistant group (85.7％ or 18/21)than in the chemo-sensitive group (53.3％ or 16/30),and the difference was statistically significant(x2=5.829,P=0.016).The higher expression of phosphorylated cofilin was also correlated with shorter PFS(x2 =21.440,P＜0.01,95％ CI:0.068-0.883),especially in the chemo-sensitive group.Conclusions Serine-3 phosphorylation status of cofilin 1 is associated with paclitaxel resistance in ovarian cancer,but the underlying mechanisms of regulation need further investigation.

Objective To study the T cells lineage polymorphism of TCR BV CDR3 in the peripheral blood of ankylosing spondylitis (AS) patients,in order to provide experimental basis for the immunological patho-genesis study of AS.Methods Twenty-six subfamilies of CDR3 T cells of TCR BV in the PBMC of AS patients were amplified by RT-PCR method,then TCR BV CDR3 lineages polymorphism were analyzed by immunization scanning spectrum.Results TCR BV CDR3 scanning spectrum of 20 active AS patients showed abnormal distribution peak,including monoclonal,oligoclonal/oligoclonal trend,skewing peak and irregular abnormal peak.Among them,some subfamilies of 18 patients showed oligoclonal/oligoclonal trend expansion,BV16 and BV18 two subfamilies of one case showed monoclonal expansion.Most spectral type of PBMC TCR BV CDR3 in five normal controls showed Gauss distribution.Conclusion TCR BV CDR3 lineage have significant characteristic polymorphism and spectrum drift characteristics in the peripheral blood of AS patients,which further indicate that T cells has plaied an important role in the immunological pathogenesis of AS.Monoclonal/oligoclonal expansion of T cells may be autoreactive T cells in nature and they may be involved in the pathogenesis of AS.

70%). TCD monitored the microembolic signals (MES) and cerebral blood flow of middle cerebral artery (MCA) using M-Mode before, after and during CAS. TCD and color ultrasound duplex were examined and new vascular events recorded during the following-up. Results Except a large numbers of MES arising from the recent contrast injection, the most MES were detected during the phase of predilation (MES number=116) and stent placement (MES number=135, 91, 113, 90, 106, 125). The MES count during the phase of postdilation was relatively reduced (MES number=8, 14, 37, 16). The most MES were characterized by high intensity and over-loading in TCD spectrum. MCA blood flow velocity varied predominantly in phases of predilation (Vsys/Vmean=46/31 to 76/54) and postdilation (Vsys/Vmean=40/26 to 74/49, 0/0 to 114/69, 35/24 to 116/71, 50/36 to 137/86). MES recorded in 2 patients prior to CAS disappeared in post-procedure. MCA blood flow velocity was enhanced or MCA pulsative index increased after CAS. No complications occurred. Tissue debris was collected in one filter device. During 3 to 19 months following-up, only one patient had a new vascular event of cerebral infarction located in the contralateral side of the CAS. Conclusions TCD monitoring might evaluate the microemboli and cerebral blood flow before, after and during CAS with filter devices. Filter devices might reduce embolization by preventing the bigger particulate microembolism into the distal part of the cerebral vessel without influencing cerebral blood flow. The gaseous and the smaller particulate microembolism unfiltrated should not result in clinical symptoms in most cases.

In adipose tissues from sreater omentum of patients with type 2 diabetes,the mRNA and protein expressions of human rearranged immunoglobulin?light chain (HSIGVL) 022 were measured by the fluorescent quantitative RT-PCR and immunohistochemistry respectively.The results showed that mRNA and protein levels of HSIGVL022 were up-regulated in patients with type 2 diabetes.The mRNA level of HSIGVL022 was linearly correlated with insulin resistance index,suggesting that this gene may play a role in the pathogenesis of insulin resistance and type 2 diabetes.

SCHWABE Company in German is the first and largest manufacturer of Ginkgo biloba preparation. The company not only has leading technology in this field, but also protects its own market effectively through the high quality of patent drafting and exactly patent layout. Based on multi-angle analysis for patent portfolio of G. biloba preparation at application time, legal status, globally layout, Chinese layout, the article provides technical reference of research and development of G. biloba, also provides valuable experience of traditonal Chinese medicine patent portfolio layout for Chinese enterprises.
Drug Industry ; economics ; legislation & jurisprudence ; trends ; Ginkgo biloba ; chemistry ; Humans ; Patents as Topic ; legislation & jurisprudence ; Phytotherapy ; economics ; trends ; Plant Preparations ; isolation & purification ; Technology, Pharmaceutical ; economics ; trends