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Ábhair agus Modhanna
3.1. Fásadh Shake Flask Cultivations
Pre-chultúr na gcealla fionraí MDCK.SUS2 i shaker flasks (fleascáin polycarbonate 125 ml Erlenmeyer, #431143, Corning®, cathair Nua-Eabhrac, NY, SAM) le toirt oibre 50 mL (wv) , i gorlann Multitron Pro (Infors HT, Bottmingen, an Eilvéis) ag 37 °C agus atmaisféar 5% CO2 le minicíocht chroitheadh 180 rpm. Trasnaíodh cealla gach 3-4 lá le dlús síolála de 0.5-0.8 × 106 cealla / mL.
IAV A/Puerto Rico/8/34 (H1N1) úsáideadh víreas síolta le haghaidh ionfhabhtú, ginte i gcealla MDCK greamaitheacha (ECACC # 84121903). Bhí titer de 1.1 × 109 TCID50 / mL. mM pyruvate), a forbraíodh go sonrach chun cealla MDCK fionraí a shaothrú [95].
Rinneadh turgnaimh fáis agus ionfhabhtuithe le haghaidh bailíochtú múnla ag baint úsáide as fleascáin chroitheadh 500 ml (#4113-0500, Nalgene™, Thermo Scientific, Waltham, MA, USA) le toirt saothraithe tosaigh de 200 ml ag 150 rpm. Rinneadh dhá shaothrú, ionfhabhtú bréige amháin (Saothrú 1), áit ar fhás cealla ar feadh thart ar 200 h, agus ionfhabhtú amháin (Saothrú 2), áit a raibh cealla ionfhabhtaithe le IAV ag thart ar 49 h tar éis ionaclaithe. Chun ionfhabhtú sioncrónach a bhaint amach sa daonra cille, baineadh úsáid as moi = 10. Mar gheall ar an tiúchan sách íseal cille tráth an ionfhabhtaithe (2.1 × 106 cill/mL) agus an moi ard a úsáideadh, ní raibh gá le hathchur meánach ná le breisiú trypsin (do ghníomhachtú víreas). Sa dá shaothrú, cuireadh uisce steiriúil Milli-Q leis roimh shampláil mar chúiteamh ar ghalú uisce (1–2 ml/lá) ó rinneadh an turgnamh i gorlann neamhhiodráitithe.
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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.