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3.1. Shake Flask Cultivations
MDCK.SUS2 suspenziya hujayralarining pre-kulturalari shaker flasks (125 ml polikarbonat Erlenmeyer flakonlari, #431143, Corning®, Nyu-York shahri, NY, AQSh) 50 ml ish hajmi (wv) bilan o'stirildi. , Multitron Pro inkubatorida (Infors HT, Bottmingen, Shveytsariya) 37 °C va 5% CO2 atmosferasida silkinish chastotasi 180 rpm. Hujayralar har 3-4 kunda 0,5-0,8 × 106 hujayra/ml urug'lik zichligi bilan o'tkazildi.
IAV A/Puerto-Riko/8/34 (H1N1) urug'lik virusi infektsiya uchun ishlatilgan, hosil qilingan. yopishgan MDCK hujayralarida (ECACC # 84121903). Urug'li virus 1,1 × 109 TCID50/ml titriga ega edi.
Hujayralar kimyoviy jihatdan aniqlangan, oqsilsiz va hayvon komponentlari bo'lmagan Smif8 (Smif8 PGD 2 ×, 5 mM glutamin va 8 bilan to'ldirilgan) muhitida o'stirilgan. mM piruvat), suspenziya MDCK hujayralarini yetishtirish uchun maxsus ishlab chiqilgan [95].
Modelni tekshirish uchun o'sish va infektsiya tajribalari 500 ml chayqaladigan kolbalar (#4113-0500, Nalgene™, Thermo Scientific, Waltham, MA, AQSh) 150 aylanish tezligida 200 ml boshlang'ich etishtirish hajmi bilan. Ikki kultivatsiya amalga oshirildi: hujayralar taxminan 200 soat davomida o'sadigan soxta infektsiya (1-kultivatsiya) va bitta infektsiya (kultivatsiya 2), hujayralar emlashdan keyin taxminan 49 soat ichida IAV bilan kasallangan. Hujayra populyatsiyasining sinxron infektsiyasiga erishish uchun moi = 10 ishlatilgan. INFEKTSION vaqtida nisbatan past hujayra konsentratsiyasi (2,1 × 106 hujayra / ml) va ishlatilgan yuqori moi tufayli, na o'rta almashtirish, na tripsin qo'shilishi (virusni faollashtirish uchun) kerak emas edi. Har ikkala kultivatsiyada suv bug'lanishini (kuniga 1-2 ml) qoplash uchun namuna olishdan oldin steril Milli-Q suv qo'shilgan, chunki tajriba gidratlanmagan inkubatorda o'tkazilgan.
Manba: https:/ /www.mdpi.com/2218-1989/12/3/239/htm
The FAI climbed 5.9 percent year-on-year in the first 11 months of 2018, quickening from the 5.7-percent growth in Jan-Oct, the National Bureau of Statistics (NBS) said Friday in an online statement.
The key indicator of investment, dubbed a major growth driver, hit the bottom in August and has since started to rebound steadily.
In the face of emerging economic challenges home and abroad, China has stepped up efforts to stabilize investment, in particular rolling out measures to motivate private investors and channel funds into infrastructure.
Friday's data showed private investment, accounting for more than 60 percent of the total FAI, expanded by a brisk 8.7 percent.
NBS spokesperson Mao Shengyong said funds into weak economic links registered rapid increases as investment in environmental protection and agriculture jumped 42 percent and 12.5 percent respectively, much faster than the average.
In breakdown, investment in high-tech and equipment manufacturing remained vigorous with 16.1-percent and 11.6-percent increases respectively in the first 11 months. Infrastructure investment gained 3.7 percent, staying flat. Investment in property development rose 9.7 percent, also unchanged.