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Objective To stimulate the expansion of natural killer cells (NK) in vivo, to achieve high expression of Flt-3 ligand ( FL) in the mouse liver by hydrodynamic injection technology , to obtain high purity of NK for cell therapy and then to purify the spleen lymphocytes by MicroBeads sorting ( MACS ) technique.Methods C57BL/6 mice were repeatedly treated with hydrodynamic injection of FL expression plasmid .Then, surface markers of splenic NK were analyzed by flow cytometry and purified by MACS technique .Results Compared with mice treated with one hydrodynamic injection, the spleen of mice treated with two hydrodynamic injections showed significant variation, with the absolute num-ber of lymphocytes increased (6.02 ±1.15) times, the proportion of NK subsets increased (2.07 ±0.39) times, and the absolute number of NK subsets increased (12.49 ±2.39) times.Cell surface marker detection confirmed that NK activity and surface markers did not change significantly .NK with a purity of about (83.81 ±0.92)％ was obtained by the combination of two magnetic beads sorted with CD 5 ( Ly-1) MicroBeads Positive selection and EasySep TM Mouse NK Cell Isolation Kit negative selection .Conclusion NK can be expanded in the spleen of mice by hydrodynamic injection of FL plasmid without influence on the NK activity and surface markers.The combined use of CD5 (Ly-1) MicroBeads positive selection with EasySep TM Mouse NK Cell Isolation Kit negative selection can effectively remove T , B and other miscellaneous cells, thereby contributing to obtaining high-purity NK. This study provides good reference for the immunotherapy of tumor cells.
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Purpose To study the relationship between autophagy and endometriosis,and its significance.Methods The eutopic and ectopic endometrial stromal cells from patients with endometriosis (n =31) and normal endometrial stromal cells (n =31) were cultured primarily and identified by immunocytochemistry.The ultrastructural changes of autophagic vacuoles were observed by transmission electron microscopy (TEM).RT-PCR and Western blot methods were used to detect LC3 mRNA and protein expression in endometrial cells of endometriosis patients.Results TEM analysis showed the number of autophagic vacuoles of eutopic and ectopic endometrial cells was fewer.RT-PCR and Western blot analysis showed that the expression of LC3 mRNA and protein in eutopic and ectopic endometrium of patients with endometriosis was significandy lower than that of the normal endometrial stromal cells (P ＜ 0.05).There was no significant difference in the expression of LC3 mRNA between eutopic and ectopic endometrium of patients with endometriosis (P ＞ 0.05),but the expression of LC3 protein in ectopic endometrium was higher than that in eutopic endometrium of patients with endometriosis (P ＜ 0.05).Conclusion Autophagy activity is downregulated in eutopic and ectopic endometrial stromal cells with endometrosis.Autophagy may be involved in the pathogenesis of endometriosis.
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<p><b>OBJECTIVE</b>To investigate the effect of thrombocytopenia on the migration patterns of adoptive dendritic cell(DC) in vivo.</p><p><b>METHODS</b>The mouse model of thrombocytopenia was established by intraperitoneal administration of anti-CD41 mAb MWReg30. Mouse bone marrow(BM)-derived DC were injected into thrombocytopenia mouse by footpad infusion and intravenous infusion. The DC migration and distribution pattern were detected by bioluminescence imaging.</p><p><b>RESULTS</b>More than 80% platelets were cleared in the experimental group which was infused with anti-CD41 antibody. At 72 h after injection, the percentage of injected DC that migrated from footpad to popliteal lymph nodes(PLNs) and inguinal lymph nodes(ILNs) were (0.32±0.02)% and (0.02±0.01)% in experimental group, and (0.27±0.15)% and (0.02±0.02)% in control group, respectively. Statistic data showed that there was no statistical difference between these 2 groups (P>0.05). The issue distribution pattern of intravenously injected DC between experimental group and control group were not distinctly different, and large amounts of injected DC accumulated in the spleen, liver draining lymph-nodes lungs and liver.</p><p><b>CONCLUSION</b>Thrombocytopenia has not a distinct effect on the migratory capacity and tissue distribution of DC by either footpad or intravenous injection.</p>
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<p><b>OBJECTIVE</b>To establish a three-dimensional image of the penile suspensory ligament, and explore a stereoscopic and multi angle observation method of patient' s penile suspensory ligament.</p><p><b>METHODS</b>This study selected the patients with small penis from our hospital as subjects. The participants were conducted on magnetic resonance imaging (MRI) examination before operation. Afterwards, the results of MRI were imported into 3D reconstruction software (MIMICS 10.0), and the suspensory ligament of penis, the pubic symphysis and other related structures were reconstructed for observation.</p><p><b>RESULTS</b>The pubic symphysis, penis and corpus spongiosum can be quite clearly displayed in the thin-section MRI images. In addition, penile suspensory with patchy distribution can be visible between lower part of ligament pubic symphysis and corpus cavernosum. Finally, we can reconstruct the three-dimensional structures through MIMICS 10.0, and then precisely describe the suspensory ligament's start-stop point, the angle with cavernous body of penis and the attached area in the corpus cavernosum penis.</p><p><b>CONCLUSION</b>Based on the MRI 3D reconstruction of deep penile suspensory ligament and adjacent structures, we can carry out dynamic, three-dimensional multi angle observation of patients deep penile suspensory ligament, and can use the reconstructed image to provide certain theory basis for the judgement of the corpus cavernosum penis extension length and penile suspensory ligament depth before penis extension operation.</p>