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Izinto kanye nezindlela
3.1. I-Shake Flask Cucultivations
Pre-cultures ye-MDCK.SUS2 amaseli amisiwe akhule ngo-shaker flasks (125 mL polycarbonate Erlenmeyer flasks, #431143, Corning®, New York city, NY, USA) anevolumu yokusebenza engu-50 mL (wv) , ku-incubator ye-Multitron Pro (Infors HT, Bottmingen, Switzerland) ku-37 °C kanye no-5% we-CO2 emkhathini onokuvama okunyakazayo okungu-180 rpm. Amaseli adluliswa njalo ezinsukwini ezi-3–4 ngobuningi bembewu obungu-0.5–0.8 × 106 amaseli/mL.
IAV A/Puerto Rico/8/34 (H1N1) igciwane lembewu lasetshenziselwa ukutheleleka, lakhiqizwa. kumaseli e-MDCK anamathelayo (ECACC # 84121903). Igciwane lembewu linesihloko esingu-1.1 × 109 TCID50/mL.
Amaseli ayetshalwa endaweni echazwe ngamakhemikhali, engenawo amaprotheni kanye nengxenye yezilwane, i-Smif8 (Smif8 PGD 2×, ehambisana no-5 mM glutamine, kanye no-8 mM pyruvate), ethuthukiswe ngokukhethekile ukutshalwa kwamaseli e-MDCK amisiwe [95].
Ukukhula nokuhlolwa kokutheleleka ukuze kuqinisekiswe imodeli kwenziwa kusetshenziswa amaflaski okunyakazisa angu-500 mL (#4113-0500, Nalgene™, Thermo Scientific, Waltham, MA, USA) enomthamo wokuqala wokutshala ongu-200 mL ngo-150 rpm. Ukulima okubili kwenziwa, ukutheleleka okukodwa okuhlekisayo (Ukulima 1), lapho amangqamuzana akhula cishe amahora angu-200, kanye nokutheleleka okukodwa (Ukuhlakulela i-2), lapho amangqamuzana atheleleke nge-IAV cishe nge-49 h post inoculation. Ukuze kuzuzwe ukutheleleka okuhambisanayo kwesibalo samaseli, i-moi = 10 yasetshenziswa. Ngenxa yokugcwala okuphansi kwamangqamuzana ngesikhathi sokutheleleka (2.1 × 106 cells/mL) kanye ne-moi ephezulu esetshenzisiwe, kwakungekho ukushintshwa okuphakathi nendawo noma ukufakwa kwe-trypsin (ukwenza igciwane kusebenze) kwakudingeka. Kuzo zombili izilimo, amanzi angenalutho e-Milli-Q engeziwe ngaphambi kokuthathwa kwesampula ukuze kunxeshezelwe ukuhwamuka kwamanzi (1–2 mL/ngosuku) njengoba ukuhlolwa kwenziwa ku-incubator engena-hydrated.
Umthombo kusuka: 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.