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Experimental teaching occupies an important position in the teaching of medical laboratory technology. At present, many experimental projects cannot be carried out due to the high cost and the long process of the experiment, the difficulty of morphology teaching and the biological safety of clinical specimens. Virtual simulation experiment platform can break through the limitations of conventional experimental teaching in time, space and safety by integrating course experiment, increasing the experiment test item, building morphological inspection data repository, the virtual operation platform for inspection instrument and equipment and the virtual experiment assessment operation platform, and play an important role in the experimental teaching. This paper reviews the current situation of experimental teaching of medical laboratory technology, the construction and application, the existing problems and countermeasures of virtual simulation experiment platform, and expects to provide reference for the construction and application of virtual simulation experiment platform for medical laboratory technology in the future.
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P2X7 receptor is an ion channel receptor with adenosine triphosphate (ATP) as its ligand, which is widely expressed in various immune cells and tissues. Activated P2X7 receptor is involved in a variety of physiological and pathological processes. P2X7 receptor is abnormally expressed in colon cancer, and plays a duel role of cancer-promoting and cancer-suppressing in colon cancer progression. When P2X7 receptor is activated by extracellular ATP, it can effectively inhibit proliferation and induce apoptosis of colon cancer cells through various mechanisms. In addition, P2X7 receptor can also promote the growth, invasion and metastasis of colon cancer. Understanding the activation of P2X7 receptor and its effect mechanism is of great significance for the treatment of colon cancer.
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Objective To obtain the PP7 bacteriophage-like particles (BLPs) carrying the polypeptide from 16 kD antigen (16kD91-110) on their surface,and evaluate their diagnostic value in tuberculosis.Methods First,the PP7 capsid protein gene containing the encoding gene of polypeptide 16kD91-110 was amplified by PCR and inserted into the plasmid pETDuet-2PP7.Then,the obtained recombinant plasmid pETDuet-2PP7-16kD91-110 was transferred into Escherichia coli,and the recombinant protein was induced and identified by SDS-PAGE and western blot.Next,the purified 2PP7-16kD91-110 BLPs were used as stimulating antigen to inject into the blood of patients with tuberculosis,and their serum antibody levels against 16 kD antigen were detected by indirect ELISA.Results The results of SDS-PAGE and transmission electron microscope showed that the 2PP7-16kD91-110 BLPs were prepared,and that the 16kD91-110 polypeptide epitopes displayed on the surface of PP7 BLPs could bind with the antibodies against 16 kD antigen specifically.After using 2PP7-16kD91-110 BLPs as antigen to stimulate the blood cells from active tuberculosis and latent tuberculosis patients,respectively,the sensitivity of interferon-γrelease assay was 75.0％ and 82.9％,respectively,in the diagnosis of active tuberculosis and latent tuberculosis,which was similar to the results of Wantai TB-IGRA (interferon-γ release assay) kits from Beijing Wantai Biological Pharmacy Enterprise Co.,Ltd..Conclusion The PP7 BLPs with the 16kD91-110 polypeptide displayed on their surface are prepared successfully,which provides a kind of safe,stable and low cost stimulating antigen for the detection of interferon-γ release,and a new method for the diagnosis of tuberculosis.
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Objective To investigate the effect of nucleus localization signal linked nucleic kinase substrate short peptide-conjugated chitosan (NNSCS)-mediated human miR-140 gene local transfection on the repair of articular cartilage defect in rabbits.Methods Eukaryotic expression plasmid GV268-miR-140 was constructed,and then negative controls GV268 and GV268-miR-140 were respectively combined with NNSCS to form NNSCS/GV268 and NNSCS/GV268-miR-140 complexes.Eighteen healthy male New Zealand white rabbits were randomly divided into transgenic group (Group A),negative control group (Group B),and sham operation group (Group C),with 6 rabbits per group.Both Groups A and B were prepared for the total cartilage damage model of femur trochlear,and Group C only exposed the articular surface of the femur trochlear.One week after operation,Group A was treated with NNS CS/GV268-miR-140 complex,Group B was given NNS CS/GV268 complex,and Group C was given equal amount of isotonic saline,twice a week for 7 weeks.The experimental animals were sacrificed at the end of the eighth week after operation.Real time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of miR-140,Sox9,Aggrecan and Hdac4 in the defect area.HE staining,safranine O/fast green staining,and Aggrecan immunohistochemical staining were used to evaluate cartilage repair in the defect area.Results RT-qPCR showed the expression of miR-140 in Group A (3.16 ± 0.37) was significantly higher than that in Group B (1 ± 0.24) and in Group C (1.24 ± 0.18) (P ＜ 0.05).The miR-140 expression in Group A obviously up-regulated the expression of SOx9 gene (4.38 ± 0.66) compared with Group B (1.04 ± 0.04) and Group C (1.19 ± 0.3),(P ＜ 0.05).The miR-140 expression in Group A obviously up-regulated the expression of Aggrecan gene (3.63 ± 0.58) (P ＜0.05) compared with Group B (1.21 ± 0.14) and Group C (1.34 ± 0.13).The miR-140 expression in Group A obviously down-regulated the expression of Hdac4 (0.37 ±0.06) compared with Group B (0.81 ± 0.06) (P ＜ 0.05).According to results of HE staining,safranine O/fast green and Aggrecan,cartilage repair was evident in Group A,while fibrous tissue proliferation and inflammatory cell infiltration were seen in the defect region in Group B,showing no cartilage repair.Conclusions NNS CS can carry exogenous genes into chondrocytes and the genes can abundantly express locally.High expression of miR-140 might significantly improve the repair of articular cartilage defect in vivoby up-regulating expressions of Aggrecan and Sox9 as well as down-regulating Hdac4 expression.
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Objective To study the effects of TRβ△ on apoptosis and proliferation of liver cancer cell line RH-35 from rat in vitro. Methods RH-35 cells were transfected by empty vector pcDNA3. 1 and expression plasmid pcDNA3. 1-TRβ△, then exposure to 10 nmol/ L T3 . RH-35 cells apoptosis and proliferation were observed by flow cytometry and MTT colorimetric assay; Levels of catenin β-1(CTNNB1), senescence marker protein-30(SMP-30) and BCL2-antagonist/ killer ( BAK ) mRNA evaluation were detected by quantitative real-time RT-PCR (RT-qPCR). Results In the presence of T3 , overexpression of TRβ△ significantly inhibited the proliferation, increased the percentage of apoptotic, down-regulated CTNNB1and SMP-30 expression, up-regulated BAK expression in RH-35 cells( P < 0. 05). Conclusion TRβ△ could inhibit the proliferation of RH-35 cells and promote their apoptosis, which may be related to upregulation of BAK genes expression and downregulation of CTNNB1 and SMP-30 gene expression, and these effects could be regulated by T3 .
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Objective To study whether osteoblast is necessary for IGF-Ⅰ to promote bone resorption by osteoclast.Methods Mouse MC3T3 osteoblast cells and mature osteoclasts induced by RANKL were cultured in vitro.These osteoblasts and osteoclasts were subjected to treatment with recombinant human insulin-like growth factor-1 (rhIGF-Ⅰ),and the activation of IGF-Ⅰ receptor was verified by Western blotting.Thereafter,osteoclasts were cultured individually or co-cultured with osteoblast,in the absence or presence of rhIGF-Ⅰ.Osteoclast proliferation and apoptosis were observed by MTT colorimetric assay and flow cytometry.Cathepsin K gene expression was detected by real-time PCR; bone adsorption activity of osteoclast was determined by resorption pits formation on calf cortex slice with toluidine blue staining.Results Western blotting result confirmed that rhIGF-Ⅰ could effectively activate IGF-Ⅰ receptors either in osteoblast or osteoclast.In co-cultured group,in the presence of rhIGF-Ⅰ osteoclast showed inhibited apoptosis,enhanced proliferation and up-regulated cathepsin K expression (P ＜ 0.05).The functional experiment revealed that osteoclasts collected from IGF-Ⅰ treated co-cultured group resulted in more resorption pits formation (P ＜ 0.05); rhIGF-Ⅰ did not show any significant effect on the individually cultured osteoclasts.Conclusion Osteoblast is necessary for osteoclast induced bone resorption resulting from IGF-Ⅰ treatment.
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To study the transactivation of a novel thyroid hormone receptor isoform, TR?. pcDNA3. 1-TR? and pGL3-Promoter/thyroid hormone responsive element were co-transfected into COS-7 cells. The expression of reporter gene was detected. It could be increased up to 45 times by T3. TR? and showed the characteristics of a transcription factor.