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Deunyddiau a Dulliau
3.1. Tyfwyd Shake Flask Cultivations
Pre-diwylliannau o gelloedd crog MDCK.SUS2 yn shaker flasks (fflasgiau polycarbonad 125 ml Erlenmeyer, #431143, Corning®, dinas Efrog Newydd, NY, UDA) gyda chyfaint gweithio 50 mL (wv) , mewn deorydd Multitron Pro (Infors HT, Bottmingen, y Swistir) ar 37 ° C a 5% CO2 awyrgylch gydag amledd ysgwyd o 180 rpm. Roedd celloedd yn cael eu pasio bob 3-4 diwrnod gyda dwysedd hadu o 0.5-0.8 × 106 o gelloedd / mL.
IAV A/Puerto Rico/8/34 (H1N1) defnyddiwyd firws hadau ar gyfer haint, a gynhyrchwyd mewn celloedd MDCK ymlynol (ECACC #84121903). Roedd gan y firws had titer o 1.1 × 109 TCID50 / mL. mM pyruvate), a ddatblygwyd yn benodol ar gyfer tyfu celloedd MDCK crog [95].
Twf ac arbrofion haint ar gyfer dilysu modelau gan ddefnyddio fflasgiau ysgwyd 500 ml (#4113-0500, Nalgene™, Thermo Scientific, Waltham, MA, UDA) gyda chyfaint amaethu cychwynnol o 200 mL ar 150 rpm. Perfformiwyd dau drin y tir, un ffug-haint (Tyfu 1), lle tyfodd celloedd am tua 200 h, ac un haint (Tyfu 2), lle cafodd celloedd eu heintio ag IAV tua 49 h ar ôl y brechiad. Er mwyn cyflawni heintiad cydamserol yn y boblogaeth gell, defnyddiwyd moi = 10. Oherwydd y crynodiad celloedd cymharol isel ar adeg yr haint (2.1 × 106 cell/mL) a'r moi uchel a ddefnyddiwyd, nid oedd angen amnewidiad canolig nac ychwanegiad trypsin (ar gyfer actifadu firws). Yn y ddau amaethiad, ychwanegwyd dŵr Milli-Q di-haint cyn samplu i wneud iawn am anweddiad dŵr (1-2 ml / dydd) ers i'r arbrawf gael ei berfformio mewn deorydd nad yw'n hydradol.
Ffynhonnell o:
https:/ /www.mdpi.com/2218-1989/12/3/239/htmhttps://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.