Preparation of vector of zinc finger protein A20 gene silence and its preliminary application / 中华创伤杂志

Objective To design and prepare an RNA interfering vector for effectively inhibiting the cellular expression of zinc finger protein A20 and observe the effect of A20 gene silence on cellular inflammatory response. Methods Specific RNA interfering oligonucleotide fragments (ASRF) were designed and synthesized artificially and the A20 RNA interfering vector pSUPER-EGFP-A20 siRNA constructed. Human monocyte cell line THP1 was used to infect the pSUPER-EGFP-A20 siRNA by means of genetic transfection technique; then, silence rate of cellular A20 was analyzed by real-time polymerase chain reaction (PCR). In the meantime, the activity of nuclear transcription factor nuclear factor-κB (NF-κB) and the level of tumor necrosis factor-α (TNF-α) in culture supernatant were measured by ELISA. Results Of two specific inhibitory oligonueleotide fragments of A20, the fragment M59465-385R/F had a higher inhibition to A20 expression, with rate of A20 gene silence of 83.86%. Preliminary application showed that after A20 gene silence, the activity of NF-κB was increased by 78.13% and the level of TNF-α in cell culture supernatant was increased by 49.30%. Conclusions Vector of A20gene silence with a high efficiency is obtained successfully. Preliminary application indicates that the expression of A20 can down-regulate the degree of cellular inflammatory responses.

Education and training for "two sides" in Laboratory Medicine / 中华医学教育探索杂志

Laboratory medicine plays an important role in health diagnosis and treatment,as a component of medicine,it is very important to pay attention to education and training for "two sides" in laboratory medicine.In the article,we discussed the concept for laboratory medicine and groups situation in clinical laboratory,the classification and the position of the"two sides"as well as training in clinical laboratory.

Exploration and Practice on Teaching Mode of Clinical Practice for the Speciality of Laboratory Medicine / 中华医学教育探索杂志

According to the characteristics of the Laboratory Medicine and our twenty years' medical educational experience ,we explore to improve the quality of education and cultivate the students' practical ability and creative ability. And we construct an educational mode for clinical practice, laboratory medical practice and research project.

The role of signal transduction molecule PLC in streptococcus pneumoniae inducing filamentous actin rearrangement in type n pneumocytes in vitro / 中华检验医学杂志

Objective To investigate whether S. pn can provoke filamentous actin (F-actin) rearrangements in vitro , which will further lead to S. pn invasion of A549 and the relationship between PLC signaling molecule and. the invasion events. Methods Labelled F-actin with FITC-phalloidin, we observed F-actin rearrangements by S. pn adhesion of type Ⅱ pneumocytes ( A549). S. pn invasion of A549 cells was determined by pretreating A549 cells with Cytochalasin D . To investigate whether F-actin rearrangements can be blocked by PLC inhibitor, A549 cells were pretreated with PLC inhibitors U73122. Results Intact S. pn can promote F-actin rearrangements. Cytochalasin D is able to prevent S. pn invasion of A549 cells. Inhibitors of PLC signal transduction molecules block F-actin rearrangements dose dependently. Conclusion S. pn can provoke F-actin rearrangements through PLC signaling pathways, which will further lead to S. pn invasion of A549 cells.

Experimental study of the fetal cells crossing the placenta into the maternal circulation / 第三军医大学学报

Objective To investigate the proof of fetal cells passing through the placental barrier into the maternal peripheral blood to provide laboratory data for the non invasive prenatal gene diagnosis of genetic diseases. Methods A total of 22 samples of placental tissues delivered(male fetus: 12, female fetus: 11) were divided into two groups for parallel section. HE staining was used to find the distribution of fetal cells in chorionic villi. In situ hybridization (ISH) technique with SRY DNA probes was used to identify the existence of fetal cells in placental villi, particularly in intervillous space. Results Light microscope examination revealed that there were fetal cells that passed through the capillary endothelium of villi and trophoblast basement membrane in the placental tissue sections of the 22 samples. ISH with SRY DNA probe also revealed that there were positive signals in the capillary of villi, at the edge of trophoblast basement membrane and in intervillous space in the placental tissue sections of the 12 placentas, but no signals were found in 10 female placentas. Conclusion This study demonstrates that the distribution of the fetal cells in the chorionic villi and intervillous space could be identified. The detection of fetal DNA in maternal circulation is one of the candidate approaches for non invasive prenatal gene diagnosis.

Attachment of Streptococcus pneumoniae to A549 cells induces tyrosine phosphorylation of host cell proteins / 中华微生物学和免疫学杂志

Objective　To investigate the potentiality of adhesion of S. pn to type Ⅱ pneumocyte (A549) in inducing tyrosine phosphorylation of A549 cell proteins and the function subcomponents involved in. Methods　Labelled S. pn with FITC for the observation of kinetic characters of adhesion in vitro; The tyrosine phosphorylation of A549 cell proteins induced by S. pn was examined by immunohistochemistry and ELISA. Results　The adhesion of S. pn to A549 cells was in a time-and dose-dependent fashion suggesting the specific interaction; Tyrosine phosphorylation induced by adhesion was dose-and time-dependent and was mediated by the surface proteins of S. pn. Conclusion　S. pn adheres to A549 cells and triggers the signal cascade in which the surface proteins of S. pn play a vital role.