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1.
Chinese Medical Sciences Journal ; (4): 179-185, 2020.
Article Dans Anglais | WPRIM | ID: wpr-828571

Résumé

Chinese scientists have been actively engaged in biotechnology research since the mid-20th century. However, biotechnology education, especially biomedical laboratory technology education, is relatively scarce in China. More and more cutting-edge equipment and techniques have been introduced into biomedical laboratories in China, but there is a lack of high-quality technicians to apply these advancements to scientific research. In addition, the traditional education and apprenticeship systems have been demonstrated little progress. To address this gap, West China Hospital of Sichuan University established a 2-year educational program for laboratory technology in 2006 based on the residency training program. The project integrates scientific methods into the research laboratory technician training in relevant disciplines, and has developed a systematic, scientific, and effective standardized training system to cultivate high-level and stable experimental technician team for the need of advanced laboratories, which has been demonstrated greatly improve the efficiency of biomedical researchers and laboratory facilities. In this article, we introduce the practical experience in establishment and development of a standardized training system for biomedical laboratory technicians to ensure the sustainable development of medical researches.

2.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 337-42, 2014.
Article Dans Anglais | WPRIM | ID: wpr-636619

Résumé

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.

3.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 337-342, 2014.
Article Dans Anglais | WPRIM | ID: wpr-351075

Résumé

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.


Sujets)
Humains , Antiviraux , Pharmacologie , Apoptose , Lignée cellulaire tumorale , Survie cellulaire , Cellules cultivées , Dependovirus , Génétique , Relation dose-effet des médicaments , Électrophorèse sur gel de polyacrylamide , Cellules épithéliales , Biologie cellulaire , Métabolisme , Ganciclovir , Pharmacologie , Régulation de l'expression des gènes tumoraux , Gènes-suicide transgéniques , Génétique , Vecteurs génétiques , Génétique , Méthode TUNEL , Luciferases , Génétique , Métabolisme , Régions promotrices (génétique) , Génétique , Alvéoles pulmonaires , Biologie cellulaire , Métabolisme , Protéine A associée au surfactant pulmonaire , Génétique , Métabolisme , Thymidine kinase , Génétique , Métabolisme
4.
Chinese Medical Journal ; (24): 3283-3288, 2013.
Article Dans Anglais | WPRIM | ID: wpr-354495

Résumé

<p><b>BACKGROUND</b>Expression of murine calcium-activated chloride channel family member 3 (mCLCA3) has been reported to be increased in the airway epithelium of asthmatic mice challenged with ovalbumin (OVA). However, its role in asthmatic airway inflammation under no OVA exposure has not yet been clarified.</p><p><b>METHODS</b>mCLCA3 plasmids were transfected into the airways of normal BALB/c mice. mCLCA3 expression and airway inflammation in mouse lung tissue were evaluated. Cell differentials and cytokines in bronchoalveolar lavage fluid (BALF) were analyzed. The expression of mCLCA3 protein and mucus protein mucin-5 subtype AC (MUC5AC) were analyzed by Western blotting. The mRNA levels of mCLCA3, MUC5AC and interleukin-13 (IL-13) were determined quantitatively.</p><p><b>RESULTS</b>mCLCA3 expression was not detected in the control group while strong immunoreactivity was detected in the OVA and mCLCA3 plasmid groups, and was strictly localized to the airway epithelium. The numbers of inflammatory cells in lung tissue and BALF were increased in both mCLCA3 plasmid and OVA groups. The protein and mRNA levels of mCLCA3 and MUC5AC in the lung tissue were significantly increased in the mCLCA3 plasmid and OVA groups compared to the control group. The level of IL-13, but not IL-4, IL-5, IFN-γ, CCL2, CCL5 or CCL11, was significantly increased compared with control group in BALF in the mCLCA3 plasmid and OVA groups. The level of IL-13 in the BALF in the mCLCA3 plasmid group was much higher than that in the OVA group (P < 0.05). The level of mCLCA3 mRNA in lung tissue was positively correlated with the levels of MUC5AC mRNA in lung tissue, IL-13 mRNA in lung tissue, the number of eosinophils in BALF, and the content of IL-13 protein in BALF. The level of IL-13 mRNA in lung tissue was positively correlated with the number of eosinophils in BALF and the level of MUC5AC mRNA in lung tissue.</p><p><b>CONCLUSION</b>These findings suggest that increased expression of a single-gene, mCLCA3, could simulate an asthma attack, and its mechanism may involve mCLCA3 overexpression up-regulating IL-13 expression.</p>


Sujets)
Animaux , Femelle , Souris , Allergènes , Asthme , Canaux chlorure , Inflammation , Métabolisme , Interleukine-13 , Métabolisme , Interleukine-4 , Génétique , Métabolisme , Interleukine-5 , Génétique , Métabolisme , Souris de lignée BALB C , Mucine-5AC , Génétique , Métabolisme , Ovalbumine , Pharmacologie
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