In 2006, the General Administration of Press and Publication of China began enforcing the first batch of the ‘China Digital Newspaper Laboratory Programs’, in which 18 national and local press groups were involved in officially unfolding the prelude of the ‘full-media convergence’, namely an intensive integration of new media into the system of traditional press. Since 2012, Chinese press industry has stepped into a ‘cold winter’. Throughout 2014 alone, the total revenue of newspaper circulation experienced a substantial decrease of 25%, with advertising revenue simultaneously decreasing by 15% (Cui and He, 2015). Several scholars suggested that vulnerable profit-making pattern, global economic recession and the decelerating growth rate of the domestic economy were the key factors for the predicament that Chinese press industry is experiencing (Zhao, 2015). However, the industry tends to ascribe the dramatic revenue decline to the prosperity of new media (cf. Cao, 2010; Zhou, 2015).
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Dàn zìwo yìshí yeyou yigèyu mou zhong lèixíng de luójí xiangguan de lilùn fangmiàn. Rénlèi sixiang bùjin fanyìngle shìjiè, érqie nénggòu genjù yiqián de xiangfa gaibiàn. Wèile dádào zhenli, yigè rén jinglì yigè tuili guòchéng, kaol? dào yiqián de shìjiàn hé lijie, yi dádào mùqián de qíngkuàng. Shèhuì biàn de yuè lái yuè fùzá, yinwèi yiqián de jingyàn yingxiangle xiànzài de yìshí, tuidòng ta cóng jiandan de lùjìng (urweg). Biànzhèng chuansuo chóngxin chuàngjiàn suo xu de tuili guòchéng, yi liaojie zài tèdìng qíngkuàng xià fashengle shénme.
Bùguò, huí dào meiguó, wo zài 2002 nián 9 yuè 8 rì shou dàole yi feng diànzi yóujiàn, nèiróng rúxià:“Wo hen yíhàn, zhidào ni de qizi shàng cì méiyou huáiyùn... Wo zài ci tígong yixie xìnxi kaol?. Shouxian, ni hé ni qizi de niánlíng dou bù xiao, huáiyùn de kenéng xìng huì hen di, dàyue shì 10%, ér ying'ér jixíng l? ye huì hen gao. Qícì, ni de jingzi de zhìliàng hé liúdòng xìng bùshì nàme hao, ni qizi de zigong nèi mó ye bùgòu hao... Dì san, ni qizi de jiànkang bù hao, you tángniàobìng hé gao xieya, huáiyùn huì èhuà zhèxie jíbìng. Shùnyì dì,IVF de fèiyòng chaoguò 20,000 yuán, chéngxù fùzá. Xiwàng yishàng xìnxi duì nín de juédìng shì youyòng de.“
Gan'en jié qián yitian, women de guanxì bàofa qilái. Lín dá xiang wèi women zuò yi dùn dà can. Wo juédé ta ye hen danxin láo la, yinwèi ta zhèngzài jieguan yigè nánrén. Wo shuole yixie da luàn lín dá de cuòwù fangshì, ta ba wo dadao zài lian shàng, wo ba ta dadaole. Ránhòu ta yòng daozi zài fángjian li zhuizhe wo. Dang wo baochí lengjìng shí, ta da diànhuà gei 911. Shèng bao lù jingchá láile, dài wo qù jianyù. Zhè míng guanyuán gàosù wo zài zhè zhong qíngkuàng xià yào dàibu zhè míng nánzi de xin zhèngcè. Suirán shèngbaoluó shì méiyou qisù, dàn zhè shì wo duì nán quán wèntí de xìngqù de kaishi.
(A–C) Immunostaining (A) and quantification (B and C) to determine endogenous poly(GR)+ punctae in control or C9-ALS iMNs with 10 nM inactive 3K3A-APC or 3K3A-APC treatment for 6 days. Quantified values represent the average number of nuclear poly(GR)+ punctae in n = 30 iMNs (controls) or 40 to 44 iMNs (C9-ALS) per line per condition from 2 control or 2 C9-ALS patient lines. For each line, iMNs were quantified from 2 independent iMN conversions per line per condition. Median ± interquartile range. Each gray circle represents the number of poly(GR)+ punctae/unit area in a single iMN. Mann-Whitney testing. Solid and dotted lines in A outline the cell body and nucleus, respectively. Scale bars: 5 μm. (D and E) Dot blot (D) and quantification (E) of poly(GR)+ levels in iMNs from 2 C9-ALS patient lines with 10 nM inactive 3K3A-APC or 3K3A-APC treatment for 6 days. Each gray circle represents 1 dot blot sample. Mean ± SD. n = 3 independent iMN conversions per line per condition. One-way ANOVA with Tukey’s correction across all comparisons. (F) Survival of control iMNs without excess glutamate with overexpression of eGFP or GR(50)-eGFP and 10 nM inactive or active 3K3A-APC. n = 90 iMNs per condition, iMNs quantified from 3 biologically independent iMN conversions. Two-sided log-rank test, corrected for multiple comparisons, statistical significance was calculated using the entire survival time course. n = 90 iMNs per condition. (G–J) Immunofluorescence analysis of total TDP-43 (G) and quantification of the ratio of nuclear to cytoplasmic TDP-43 in control, C9-ALS (H), or sporadic ALS iMNs (I and J). Ratio of nuclear to cytoplasmic TDP-43 in individual C9-ALS iMNs treated with 10 nM inactive 3K3A-APC or 3K3A-APC for 6 days. iMNs from 2 controls, 2 C9-ALS, and 4 sporadic ALS patients were quantified. n = 30 iMNs per line (control and C9-ALS) per condition or n = 26 iMNs (I), 30 iMNs (J) (inactive 3K3A-APC), or n = 35 iMNs (J) (3K3A-APC) (sporadic ALS) per condition per line from 2 biologically independent iMN conversions were quantified. Each gray circle represents a single iMN. For H, median ± interquartile range. Kruskal-Wallis testing. For I and J, mean ± SEM. Unpaired t test with Welch’s correction. Scale bars: 5 μm. Dotted lines outline the nucleus and cell body. The day of differentiation stated on each panel indicates the day of differentiation on which the experimental treatment or time course was initiated.
The cDNA for each iMN transcription factor or the mRFP-GFP-LC3 construct was subcloned into the pMXs retroviral expression vector (4). The Hb9::RFP lentiviral vector was previously purchased from Addgene (ID: 37081). Viruses were produced as follows. HEK 293T cells (ATCC, CRL-11268) were transfected at 80% to 90% confluence with viral vectors containing genes of interest and viral packaging plasmids (PIK-MLV-gp and pHDM for retrovirus, pPAX2 and VSVG for lentivirus) using polyethylenimine (Sigma-Aldrich). HEK 293T were tested for mycoplasma before, during, and after the study and were negative. Viruses were harvested at 48 hours and 72 hours after transfection. Viral supernatants were filtered with 0.45-μm filters, incubated with Lenti-X concentrator (Clontech) for 24 hours at 4°C, and centrifuged at 1500 g at 4°C for 45 minutes. Pellets were resuspended in 300 μL DMEM plus 10% FBS and stored at –80°C.
Jíshi shèhuì wendìng, wénmíng ye keyi gaibiàn. Wénmíng de benzhí huò xìngzhì shì yu zhurú chuàngzào yidìng gonggòng kongjian de xiezuò jìshù xiang liánxì de yìshí móshì. Guangyì shàng de zongjiào dìngyìle ta de línghún. Youshí (rú mùqián) wénmíng zài zhèngzhì hé shèhuì jiégòu bù biàn shí fasheng biànhuà. Youshí (dang luóma lúnxiàn shí) wénmíng zài bùduàn biànhuà de shèhuì zhong shengcún. Zhè ben shu jiang guanzhù zài yìnshua wénhuà hé diànzi yúlè wénhuà zhi jian fasheng de jiàzhíguan de zhuanbiàn. Ta hái jiang guanzhù zhichí mei yi tào liniàn de tongxìn jìshù.