Application of Mimics Medical 21.0 Software in thoracoscopic anatomical sublobectomy.

BACKGROUND: The anatomical structure of pulmonary segments is complex, and there are many anatomical variations, making the operation more difficult, so we used Mimics Medical 21.0 software for three-dimensional computed tomography bronchography and angiography (3D-CTBA), carefully formulated the surgical plan and on this basis completed the video-assisted thoracoscopic anatomical sublobectomy. METHODS: A total of 38 patients with pulmonary nodules were selected and received video-assisted thoracoscopic anatomical sublobectomy after using Mimics Medica 21.0 software for 3D-CTBA. RESULTS: The mean operative duration was 158.42±20.21 minutes, and the operative hemorrhage was 97.66±22.37 mL. In pathological diagnoses, there were 5 benign cases (13.2%), 9 cases with atypical adenomatous hyperplasia (23.7%), 12 cases with adenocarcinoma in situ (31.6%), 11 cases with minimally invasive adenocarcinoma (28.9%) and 1 case with invasive adenocarcinoma (2.6%); 8.29±0.98 lymph nodes sampled had no metastasis. The chest tube drainage duration and postoperative hospital stay were 2.47±0.73 days and 5.47±0.73 days, respectively. CONCLUSIONS: Using Mimics Medical 21.0 software can quickly and accurately complete 3D-CTBA, which is beneficial to formulate a personalized anatomical sublobectomy surgical plan.

Astragalus polysaccharide stimulates glucose uptake in L6 myotubes through AMPK activation and AS160/TBC1D4 phosphorylation.

AIM: To establish the mechanism responsible for the stimulation of glucose uptake by Astragalus polysaccharide (APS), extracted from Astragalus membranaceus Bunge, in L6 myotubes in vitro. METHODS: APS-stimulated glucose uptake in L6 myotubes was measured using the 2-deoxy-[(3)H]-D-glucose method. The adenine nucleotide contents in the cells were measured by HPLC. The phosphorylation of AMP-activated protein kinase (AMPK) and Akt substrate of 160 kDa (AS160) was examined using Western blot analysis. The cells transfected with 4P mutant AS160 (AS160-4P) were constructed using gene transfer approach. RESULTS: Treatment of L6 myotubes with APS (100-1600 µg/mL) significantly increased glucose uptake in time- and concentration-dependent manners. The maximal glucose uptake was reached in the cells treated with APS (400 µg/mL) for 36 h. The APS-stimulated glucose uptake was significantly attenuated by pretreatment with Compound C, a selective AMPK inhibitor or in the cells overexpressing AS160-4P. Treatment of L6 myotubes with APS strongly promoted the activation of AMPK. We further demonstrated that either Ca(2+)/calmodulin-dependent protein kinase kinase ß (CaMKKß) or liver kinase B1 (LKB1) mediated APS-induced activation of AMPK in L6 myotubes, and the increased cellular AMP: ATP ratio was also involved. Treatment of L6 myotubes with APS robustly enhanced the phosphorylation of AS160, which was significantly attenuated by pretreatment with Compound C. CONCLUSION: Our results demonstrate that APS stimulates glucose uptake in L6 myotubes through the AMP-AMPK-AS160 pathway, which may contribute to its hypoglycemic effect.

[Construction of a non-viral vector H1s-EGFc and a preliminary study on its function].

OBJECTIVE: To construct a non-viral vector for targeting cancer gene therapy. METHODS: The coding sequence of H1s-EGFc was inserted into the expression vectors of Pichia pastoris, and the fusion protein was expressed in secretary way. H1s-EGFc was purified by anion exchange chromatography and size exclusion chromatography. H1s-EGFc fusion protein and "killing gene" expression recombinant pKG plasmid DNA were dissolved in serum-free RPMI-1640 culture to produce H1s-EGFc/pKG complex. HeLa cells, an epidermal growth factor receptor (EGFR) highly expressing cell line, and Jurkat cells, an EGFR non-expressing cell line, were cultured and transfected with H1s-EGFc/pKG complex of different concentrations. Trypan blue staining was used to calculate the number of live cells and the killing rate of H1s-EGFc/pKG. RESULTS: H1s-EGFc fusion protein was constructed and expressed with a purity of over 90%. When the concentrations of H1s-EGFc/pKG complex were 3 microg/ml, 6 microg/ml, and 9 microg/ml respectively the killing rates were 30.6%, 36.2%, and 58.1% respectively. CONCLUSION: The fusion protein H1s-EGFc binds functional gene efficiently and targets it into specific cells. It can be used as non-viral vector in target cancer gene therapy.