Xiaohua Chen

Ethical considerations constrain the in vivo study of human hemopoietic stem cells (HSC). To overcome this limitation, small animal models of human HSC engraftment have been used. We report the development and characterization of a new genetic stock of IL-2R common gamma-chain deficient NOD/LtSz-scid (NOD-scid IL2Rgamma(null)) mice and document their ability to support human mobilized blood HSC engraftment and multilineage differentiation. NOD-scid IL2Rgamma(null) mice are deficient in mature lymphocytes and NK cells, survive beyond 16 mo of age, and even after sublethal irradiation resist lymphoma development. Engraftment of NOD-scid IL2Rgamma(null) mice with human HSC generate 6-fold higher percentages of human CD45(+) cells in host bone marrow than with similarly treated NOD-scid mice. These human cells include B cells, NK cells, myeloid cells, plasmacytoid dendritic cells, and HSC. Spleens from engrafted NOD-scid IL2Rgamma(null) mice contain human Ig(+) B cells and lower numbers of human CD3(+) T cells. Coadministration of human Fc-IL7 fusion protein results in high percentages of human CD4(+)CD8(+) thymocytes as well human CD4(+)CD8(-) and CD4(-)CD8(+) peripheral blood and splenic T cells. De novo human T cell development in NOD-scid IL2Rgamma(null) mice was validated by 1) high levels of TCR excision circles, 2) complex TCRbeta repertoire diversity, and 3) proliferative responses to PHA and streptococcal superantigen, streptococcal pyrogenic exotoxin. Thus, NOD-scid IL2Rgamma(null) mice engrafted with human mobilized blood stem cells provide a new in vivo long-lived model of robust multilineage human HSC engraftment.

BACKGROUND: Physical therapy can prevent functional impairments and improve the quality of life of patients after hospital discharge. However, the effect of early mobilization on patients with a critical illness remains unclear. This study was performed to assess the evidence available regarding the effect of early mobilization on critically ill patients in the intensive care unit (ICU). METHODS: Electronic databases were searched from their inception to March 21, 2019. Randomized controlled trials (RCTs) comprising critically ill patients who received early mobilization were included. The methodological quality and risk of bias of each eligible trial were assessed using the Cochrane Collaboration tool. Data were extracted using a standard collection form each included study, and processed using the Mantel-Haenszel (M-H) or inverse-variance (I-V) test in the STATA v12.0 statistical software. RESULTS: A total of 1,898 records were screened. Twenty-three RCTs comprising 2,308 critically ill patients were ultimately included. Early mobilization decreased the incidence of ICU-acquired weakness (ICU-AW) at hospital discharge (three studies, 190 patients, relative risk (RR): 0.60, 95% confidence interval (CI) [0.40, 0.90]; p = 0.013, I2 = 0.0%), increased the number of patients who were able to stand (one study, 50 patients, 90% vs. 62%, p = 0.02), increased the number of ventilator-free days (six studies, 745 patients, standardized mean difference (SMD): 0.17, 95% CI [0.02, 0.31]; p = 0.023, I2 = 35.5%) during hospitalization, increased the distance the patient was able to walk unassisted (one study, 104 patients, 33.4 (0-91.4) meters vs. 0 (0-30.4) meters, p = 0.004) at hospital discharge, and increased the discharged-to-home rate (seven studies, 793 patients, RR: 1.16, 95% CI [1.00, 1.34]; p = 0.046). The mortality (28-day, ICU and hospital) and adverse event rates were moderately increased by early mobilization, but the differences were statistically non-significant. However, due to the substantial heterogeneity among the included studies, and the low quality of the evidence, the results of this study should be interpreted with caution. Publication bias was not identified. CONCLUSIONS: Early mobilization appears to decrease the incidence of ICU-AW, improve the functional capacity, and increase the number of ventilator-free days and the discharged-to-home rate for patients with a critical illness in the ICU setting.

It has long been assumed that the C-terminal motif, NPXY, is the internalization signal for β-amyloid precursor protein (APP) and that the NPXY tyrosine (Tyr743 by APP751 numbering, Tyr682 in APP695) is required for APP endocytosis. To evaluate this tenet and to identify the specific amino acids subserving APP endocytosis, we mutated all tyrosines in the APP cytoplasmic domain and amino acids within the sequence GYENPTY (amino acids 737–743). Stable cell lines expressing these mutations were assessed for APP endocytosis, secretion, and turnover. Normal APP endocytosis was observed for cells expressing Y709A, G737A, and Y743A mutations. However, Y738A, N740A, and P741A or the double mutation of Y738A/P741A significantly impaired APP internalization to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic domain (ΔC), arguing that the dominant signal for APP endocytosis is the tetrapeptide YENP. Although not an APP internalization signal, Tyr743 regulates rapid APP turnover because half-life increased by 50% with the Y743A mutation alone. Secretion of the APP-derived proteolytic fragment, Aβ, was tightly correlated with APP internalization, such that Aβ secretion was unchanged for cells having normal APP endocytosis but significantly decreased for endocytosis-deficient cell lines. Remarkably, secretion of the Aβ42 isoform was also reduced in parallel with endocytosis from internalization-deficient cell lines, suggesting an important role for APP endocytosis in the secretion of this highly pathogenic Aβ species. It has long been assumed that the C-terminal motif, NPXY, is the internalization signal for β-amyloid precursor protein (APP) and that the NPXY tyrosine (Tyr743 by APP751 numbering, Tyr682 in APP695) is required for APP endocytosis. To evaluate this tenet and to identify the specific amino acids subserving APP endocytosis, we mutated all tyrosines in the APP cytoplasmic domain and amino acids within the sequence GYENPTY (amino acids 737–743). Stable cell lines expressing these mutations were assessed for APP endocytosis, secretion, and turnover. Normal APP endocytosis was observed for cells expressing Y709A, G737A, and Y743A mutations. However, Y738A, N740A, and P741A or the double mutation of Y738A/P741A significantly impaired APP internalization to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic domain (ΔC), arguing that the dominant signal for APP endocytosis is the tetrapeptide YENP. Although not an APP internalization signal, Tyr743 regulates rapid APP turnover because half-life increased by 50% with the Y743A mutation alone. Secretion of the APP-derived proteolytic fragment, Aβ, was tightly correlated with APP internalization, such that Aβ secretion was unchanged for cells having normal APP endocytosis but significantly decreased for endocytosis-deficient cell lines. Remarkably, secretion of the Aβ42 isoform was also reduced in parallel with endocytosis from internalization-deficient cell lines, suggesting an important role for APP endocytosis in the secretion of this highly pathogenic Aβ species. β-amyloid precursor protein Alzheimer's disease low density lipoprotein receptor Chinese hamster ovary wild type N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine enzyme-linked immunosorbent assay analysis of variance. APPs, secreted N1-terminal ectodomain of APP APP1 is a transmembrane protein with homology to glycosylated cell surface receptors (1Kang J. Lemaire H.-G. Unterbeck A. Salbaum J.M. Masters C.L. Grzeschik K.-H. Multhaup G. Beyreuther K. Muller-Hill B. Nature. 1987; 325: 733-736Crossref PubMed Scopus (3916) Google Scholar), can reside at the cell surface (2Shivers B.D. Hilbich C. Multhaup G. Salbaum M. Beyreuther K. Seeburg P.H. EMBO J. 1988; 7: 1365-1370Crossref PubMed Scopus (387) Google Scholar, 3Breen K. Bruce M. Anderson B. J. Neurosci. Res. 1991; 28: 90-100Crossref PubMed Scopus (221) Google Scholar, 4Jung S.S. Nalbantoglu J. Cashman N.R. J Neurosci. Res. 1996; 46: 336-348Crossref PubMed Scopus (42) Google Scholar) and is reinternalized via clathrin-coated pits (5Nordstedt C. Caporaso G.L. Thyberg J. Gandy S.E. Greengard P. J. Biol. Chem. 1993; 268: 608-612Abstract PubMed Google Scholar, J. 1996; PubMed Google Scholar) to the C. A. Nature. PubMed Scopus Google Scholar, PubMed Scopus Google APP to to the cell surface J. Biol. PubMed Scopus Google Scholar, J. 1996; PubMed Google However, APP can also secreted the secreted APP ectodomain and Aβ, the protein of in Alzheimer's disease in PubMed Scopus Google Aβ to the by Aβ is from the precursor is an important of of Aβ, by amino acids at the and from cells normal C. C. A. Nature. PubMed Scopus Google Scholar, M. J. B. PubMed Scopus Google Scholar, C. M. M. J. M. M. P. G. 1996; PubMed Scopus Google in in 1993; PubMed Scopus Google Scholar) to pathogenic and a for in with Biol. 1996; Scholar), with type P. J. M. G. M. 1996; PubMed Scopus Google Scholar), and 1996; PubMed Scopus Google of Aβ in is However, cell lines expressing wild type APP can and Aβ internalization of APP from the cell surface J. Biol. Chem. PubMed Google Scholar, J. Biol. Chem. 1996; PubMed Scopus Google Although mutations in APP can Aβ secretion the J. Biol. Chem. 1996; PubMed Scopus Google Scholar, M. C. K. P. C. Nature. PubMed Scopus Google Scholar, 1993; PubMed Scopus Google Scholar, PubMed Scopus Google Scholar), all wild type the for Aβ to of APP from the cell To the specific of APP to Aβ42 in has not been endocytosis in the cytoplasmic C-terminal NPXY sequence similar to that in the of the low density lipoprotein receptor and the protein J. EMBO J. 1988; 7: PubMed Scopus Google Scholar) is also in the tyrosine in NPXY is for endocytosis J. Biol. Chem. PubMed Google Scholar, Anderson PubMed Scopus Google Scholar), has long been assumed that the tyrosine in the APP NPXY is the signal for APP endocytosis. the an amino of the NPXY was a to endocytosis, the internalization in Biol. 1993; PubMed Scopus Google the to the APP signal from an receptor was to the GYENPTY (amino acids with a by tyrosine in the to the A. J. Biol. Chem. PubMed Scopus Google However, because the receptor the and the of the APP signal with to the is this to of the in that the of the signal Aβ42 in has not been to identify the specific amino acids required for APP endocytosis and to APP tyrosines with endocytosis of J. A. PubMed Scopus Google Scholar, B. A. M. A. PubMed Scopus Google Scholar), we mutations of all the APP cytoplasmic domain tyrosines of the amino acids and in the GYENPTY the of these mutations in cell lines and that amino acids to APP endocytosis and the of APP secreted we observed that the tyrosine in to APP endocytosis, the tyrosine in this the were Y743A cell lines APP turnover. we a the of APP endocytosis and Aβ in Aβ42 and Aβ secretion suggesting that to the secretion of the pathogenic Aβ42 and the by cell cells been in to APP to cells C. A. Nature. PubMed Scopus Google B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar, P. C. M. M. J. C. Nature. PubMed Scopus Google Scholar, M. C. C. J. B. A. PubMed Scopus Google Scholar) and a for APP and J. 1996; PubMed Google Scholar, J. 1996; PubMed Google Scholar, J. Biol. Chem. PubMed Google Scholar, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and of cells expressing APP or APP with C-terminal mutations and APP and we the tetrapeptide motif, (amino acids in APP751 or in APP695) in the APP the dominant APP we observed that mutation of Tyr743 in the NPXY APP endocytosis, this mutation significantly of we and parallel the secretion of APPs, Aβ, and Aβ42 in with C-terminal mutations that normal APP APP cell surface is in clathrin-coated (5Nordstedt C. Caporaso G.L. Thyberg J. Gandy S.E. Greengard P. J. Biol. Chem. 1993; 268: 608-612Abstract PubMed Google Scholar, J. 1996; PubMed Google Scholar), that APP a and a to cell surface receptors in Biol. 1993; PubMed Scopus Google receptor to is the role in endocytosis, NPXY has been assumed to the signal for APP endocytosis J. Biol. Chem. PubMed Google to the amino acids of this NPXY motif, an amino of NPXY also to endocytosis, the signal for this the J. Biol. Chem. PubMed Google been the motif, an sequence in the APP the for APP endocytosis and J. Biol. Chem. PubMed Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and C. A. J. Biol. PubMed Scopus Google Scholar, B. K. B. J. Biol. Chem. PubMed Scopus Google Although that the APP signal was within this motif, the specific amino acids APP endocytosis to endocytosis also been for the tyrosine the and for the in GYENPTY of APP A. J. Biol. Chem. PubMed Scopus Google we the to to APP endocytosis a for APP endocytosis J. 1996; PubMed Google we that mutations of or or a double mutation of and but not a mutation of the C-terminal in significantly APP endocytosis. mutations within the all reduced APP internalization that the amino is the signal for APP endocytosis. amino to a of the APP signal because the mutation reduced endocytosis to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic that to APP internalization in suggesting that these amino Tyr743 in NPXY, not of the APP amino acids from the amino acids from the was the in the A. J. Biol. Chem. PubMed Scopus Google Scholar), and this the tyrosine internalization in the Although ectodomain of APP a role in in cells B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar), for the tyrosine subserving of APP has also been C. A. J. Biol. PubMed Scopus Google It that the tyrosine in APP751 and in APP695) with the protein with homology that with to APP to the in cells P. J. A. PubMed Scopus Google the role of Tyr743 in APP we that Tyr743 in the NPXY is not an APP signal and that cells APP with this mutation normal of Although endocytosis was not in cells with mutations of the NPXY tyrosine also observed normal Aβ secretion from cell lines J. J. Biol. Chem. PubMed Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Although not an signal, reduced the rapid turnover of Although the APP half-life was in Y743A APP was from to glycosylated APP to and suggesting that the was normal for these cell lines. we that Tyr743 in a signal for that cells expressing C-terminal reduced of Aβ secretion J. Biol. Chem. PubMed Google Scholar, K. B. M. Alzheimer's and and Aβ was by cells expressing mutations that reduced APP endocytosis. Y738A, N740A, and cell lines all reduced Aβ secretion by in Aβ secretion was observed for Y709A, G737A, or cell lines that also normal APP endocytosis. the of the for the of Aβ by we Aβ42 secretion by we were to that in parallel with the of Aβ secretion, the Y738A, N740A, and cell lines also secreted that of APP in the to Aβ42 However, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, B. J. A. PubMed Scopus Google Scholar, G. J. M. P. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google Scholar) and cells J. Biol. Chem. 1996; PubMed Scopus Google Scholar, B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar) to Aβ42 in the of the C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, M. A. Multhaup G. Masters C.L. Beyreuther K. A. PubMed Scopus Google Scholar, B. Masters C.L. K. Beyreuther K. PubMed Scopus Google Scholar), we were to that to significantly to the of Aβ42 secreted from cell lines. However, is not the Aβ in the secreted by the that Aβ42 not because the level of Aβ42 the was the we not Aβ in for Aβ, and Aβ42 in cell or in or of C-terminal required to or not a in endocytosis can the Aβ species. that that mutations that APP internalization reduced Aβ secretion, the mutations APP to is the amino acids in the endocytosis, is to that by with such or in G. P. PubMed Scopus Google in APP was assessed in cells or M. B. J. Biol. Chem. PubMed Scopus Google Scholar, J. A. Scholar) that the of APP with these in and that Aβ is by of this to APP in the because of reduced by M. B. J. Biol. Chem. PubMed Scopus Google Scholar), because of reduced of APP at the cell with in an of the because of in cells increased secretion J. A. Scholar), an to of APP at or the cell for cells Greengard P. J. Biol. Chem. PubMed Scopus Google of with APP to within B.D. A. J. Biol. Chem. PubMed Scopus Google an NPXY APP and the receptor protein by of the and has also been that APP and M. B. J. J. Biol. Chem. PubMed Scopus Google that the dominant signal for APP endocytosis in the that in the APP to APP and that of APP to and Aβ of the by cells APP secreted for or the of of Aβ and secretion for APP1 is a transmembrane protein with homology to glycosylated cell surface receptors (1Kang J. Lemaire H.-G. Unterbeck A. Salbaum J.M. Masters C.L. Grzeschik K.-H. Multhaup G. Beyreuther K. Muller-Hill B. Nature. 1987; 325: 733-736Crossref PubMed Scopus (3916) Google Scholar), can reside at the cell surface (2Shivers B.D. Hilbich C. Multhaup G. Salbaum M. Beyreuther K. Seeburg P.H. EMBO J. 1988; 7: 1365-1370Crossref PubMed Scopus (387) Google Scholar, 3Breen K. Bruce M. Anderson B. J. Neurosci. Res. 1991; 28: 90-100Crossref PubMed Scopus (221) Google Scholar, 4Jung S.S. Nalbantoglu J. Cashman N.R. J Neurosci. Res. 1996; 46: 336-348Crossref PubMed Scopus (42) Google Scholar) and is reinternalized via clathrin-coated pits (5Nordstedt C. Caporaso G.L. Thyberg J. Gandy S.E. Greengard P. J. Biol. Chem. 1993; 268: 608-612Abstract PubMed Google Scholar, J. 1996; PubMed Google Scholar) to the C. A. Nature. PubMed Scopus Google Scholar, PubMed Scopus Google APP to to the cell surface J. Biol. PubMed Scopus Google Scholar, J. 1996; PubMed Google However, APP can also secreted the secreted APP ectodomain and Aβ, the protein of in Alzheimer's disease in PubMed Scopus Google Aβ to the by Aβ is from the precursor is an important of of Aβ, by amino acids at the and from cells normal C. C. A. Nature. PubMed Scopus Google Scholar, M. J. B. PubMed Scopus Google Scholar, C. M. M. J. M. M. P. G. 1996; PubMed Scopus Google in in 1993; PubMed Scopus Google Scholar) to pathogenic and a for in with Biol. 1996; Scholar), with type P. J. M. G. M. 1996; PubMed Scopus Google Scholar), and 1996; PubMed Scopus Google of Aβ in is However, cell lines expressing wild type APP can and Aβ internalization of APP from the cell surface J. Biol. Chem. PubMed Google Scholar, J. Biol. Chem. 1996; PubMed Scopus Google Although mutations in APP can Aβ secretion the J. Biol. Chem. 1996; PubMed Scopus Google Scholar, M. C. K. P. C. Nature. PubMed Scopus Google Scholar, 1993; PubMed Scopus Google Scholar, PubMed Scopus Google Scholar), all wild type the for Aβ to of APP from the cell To the specific of APP to Aβ42 in has not been APP endocytosis in the cytoplasmic C-terminal NPXY sequence similar to that in the of the low density lipoprotein receptor and the protein J. EMBO J. 1988; 7: PubMed Scopus Google Scholar) is also in the tyrosine in NPXY is for endocytosis J. Biol. Chem. PubMed Google Scholar, Anderson PubMed Scopus Google Scholar), has long been assumed that the tyrosine in the APP NPXY is the signal for APP endocytosis. the an amino of the NPXY was a to endocytosis, the internalization in Biol. 1993; PubMed Scopus Google the to the APP signal from an receptor was to the GYENPTY (amino acids with a by tyrosine in the to the A. J. Biol. Chem. PubMed Scopus Google However, because the receptor the and the of the APP signal with to the is this to of the in that the of the signal Aβ42 in has not been to identify the specific amino acids required for APP endocytosis and to APP tyrosines with endocytosis of J. A. PubMed Scopus Google Scholar, B. A. M. A. PubMed Scopus Google Scholar), we mutations of all the APP cytoplasmic domain tyrosines of the amino acids and in the GYENPTY the of these mutations in cell lines and that amino acids to APP endocytosis and the of APP secreted we observed that the tyrosine in to APP endocytosis, the tyrosine in this the were Y743A cell lines APP turnover. we a the of APP endocytosis and Aβ in Aβ42 and Aβ secretion suggesting that to the secretion of the pathogenic Aβ42 and the by cell lines. cells been in to APP to cells C. A. Nature. PubMed Scopus Google B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar, P. C. M. M. J. C. Nature. PubMed Scopus Google Scholar, M. C. C. J. B. A. PubMed Scopus Google Scholar) and a for APP and J. 1996; PubMed Google Scholar, J. 1996; PubMed Google Scholar, J. Biol. Chem. PubMed Google Scholar, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and of cells expressing APP or APP with C-terminal mutations and APP and we the tetrapeptide motif, (amino acids in APP751 or in APP695) in the APP the dominant APP we observed that mutation of Tyr743 in the NPXY APP endocytosis, this mutation significantly of we and parallel the secretion of APPs, Aβ, and Aβ42 in with C-terminal mutations that normal APP APP cell surface is in clathrin-coated (5Nordstedt C. Caporaso G.L. Thyberg J. Gandy S.E. Greengard P. J. Biol. Chem. 1993; 268: 608-612Abstract PubMed Google Scholar, J. 1996; PubMed Google Scholar), that APP a and a to cell surface receptors in Biol. 1993; PubMed Scopus Google receptor to is the role in endocytosis, NPXY has been assumed to the signal for APP endocytosis J. Biol. Chem. PubMed Google to the amino acids of this NPXY motif, an amino of NPXY also to endocytosis, the signal for this the J. Biol. Chem. PubMed Google been the motif, an sequence in the APP the for APP endocytosis and J. Biol. Chem. PubMed Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and C. A. J. Biol. PubMed Scopus Google Scholar, B. K. B. J. Biol. Chem. PubMed Scopus Google Although that the APP signal was within this motif, the specific amino acids APP endocytosis to endocytosis also been for the tyrosine the and for the in GYENPTY of APP A. J. Biol. Chem. PubMed Scopus Google we the to to APP endocytosis a for APP endocytosis J. 1996; PubMed Google we that mutations of or or a double mutation of and but not a mutation of the C-terminal in significantly APP endocytosis. mutations within the all reduced APP internalization that the amino is the signal for APP endocytosis. amino to a of the APP signal because the mutation reduced endocytosis to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic that to APP internalization in suggesting that these amino Tyr743 in NPXY, not of the APP amino acids from the amino acids from the was the in the A. J. Biol. Chem. PubMed Scopus Google Scholar), and this the tyrosine internalization in the Although ectodomain of APP a role in in cells B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar), for the tyrosine subserving of APP has also been C. A. J. Biol. PubMed Scopus Google It that the tyrosine in APP751 and in APP695) with the protein with homology that with to APP to the in cells P. J. A. PubMed Scopus Google the role of Tyr743 in APP we that Tyr743 in the NPXY is not an APP signal and that cells APP with this mutation normal of Although endocytosis was not in cells with mutations of the NPXY tyrosine also observed normal Aβ secretion from cell lines J. J. Biol. Chem. PubMed Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Although not an signal, reduced the rapid turnover of Although the APP half-life was in Y743A APP was from to glycosylated APP to and suggesting that the was normal for these cell lines. we that Tyr743 in a signal for that cells expressing C-terminal reduced of Aβ secretion J. Biol. Chem. PubMed Google Scholar, K. B. M. Alzheimer's and and Aβ was by cells expressing mutations that reduced APP endocytosis. Y738A, N740A, and cell lines all reduced Aβ secretion by in Aβ secretion was observed for Y709A, G737A, or cell lines that also normal APP endocytosis. the of the for the of Aβ by we Aβ42 secretion by we were to that in parallel with the of Aβ secretion, the Y738A, N740A, and cell lines also secreted that of APP in the to Aβ42 However, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, B. J. A. PubMed Scopus Google Scholar, G. J. M. P. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google Scholar) and cells J. Biol. Chem. 1996; PubMed Scopus Google Scholar, B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar) to Aβ42 in the of the C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, M. A. Multhaup G. Masters C.L. Beyreuther K. A. PubMed Scopus Google Scholar, B. Masters C.L. K. Beyreuther K. PubMed Scopus Google Scholar), we were to that to significantly to the of Aβ42 secreted from cell lines. However, is not the Aβ in the secreted by the that Aβ42 not because the level of Aβ42 the was the we not Aβ in for Aβ, and Aβ42 in cell or in or of C-terminal required to or not a in endocytosis can the Aβ species. that that mutations that APP internalization reduced Aβ secretion, the mutations APP to is the amino acids in the endocytosis, is to that by with such or in G. P. PubMed Scopus Google in APP was assessed in cells or M. B. J. Biol. Chem. PubMed Scopus Google Scholar, J. A. Scholar) that the of APP with these in and that Aβ is by of this to APP in the because of reduced by M. B. J. Biol. Chem. PubMed Scopus Google Scholar), because of reduced of APP at the cell with in an of the because of in cells increased secretion J. A. Scholar), an to of APP at or the cell for cells Greengard P. J. Biol. Chem. PubMed Scopus Google of with APP to within B.D. A. J. Biol. Chem. PubMed Scopus Google an NPXY APP and the receptor protein by of the and has also been that APP and M. B. J. J. Biol. Chem. PubMed Scopus Google that the dominant signal for APP endocytosis in the that in the APP to APP and that of APP to and Aβ of the by cells APP secreted for or the of of Aβ and secretion for cells been in to APP to cells C. A. Nature. PubMed Scopus Google B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar, P. C. M. M. J. C. Nature. PubMed Scopus Google Scholar, M. C. C. J. B. A. PubMed Scopus Google Scholar) and a for APP and J. 1996; PubMed Google Scholar, J. 1996; PubMed Google Scholar, J. Biol. Chem. PubMed Google Scholar, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and of cells expressing APP or APP with C-terminal mutations and APP and we the tetrapeptide motif, (amino acids in APP751 or in APP695) in the APP the dominant APP we observed that mutation of Tyr743 in the NPXY APP endocytosis, this mutation significantly of we and parallel the secretion of APPs, Aβ, and Aβ42 in with C-terminal mutations that normal APP endocytosis. APP cell surface is in clathrin-coated (5Nordstedt C. Caporaso G.L. Thyberg J. Gandy S.E. Greengard P. J. Biol. Chem. 1993; 268: 608-612Abstract PubMed Google Scholar, J. 1996; PubMed Google Scholar), that APP a and a to cell surface receptors in Biol. 1993; PubMed Scopus Google receptor to is the role in endocytosis, NPXY has been assumed to the signal for APP endocytosis J. Biol. Chem. PubMed Google to the amino acids of this NPXY motif, an amino of NPXY also to endocytosis, the signal for this the J. Biol. Chem. PubMed Google been the motif, an sequence in the APP the for APP endocytosis and J. Biol. Chem. PubMed Google Scholar, A. J. Biol. Chem. PubMed Scopus Google Scholar, K. B. M. Alzheimer's and and C. A. J. Biol. PubMed Scopus Google Scholar, B. K. B. J. Biol. Chem. PubMed Scopus Google Although that the APP signal was within this motif, the specific amino acids APP endocytosis to endocytosis also been for the tyrosine the and for the in GYENPTY of APP A. J. Biol. Chem. PubMed Scopus Google we the to to APP endocytosis a for APP endocytosis J. 1996; PubMed Google we that mutations of or or a double mutation of and but not a mutation of the C-terminal in significantly APP endocytosis. mutations within the all reduced APP internalization that the amino is the signal for APP endocytosis. amino to a of the APP signal because the mutation reduced endocytosis to a level similar to that observed for cells lacking nearly the entire APP cytoplasmic that to APP internalization in suggesting that these amino Tyr743 in NPXY, not of the APP amino acids from the amino acids from the was the in the A. J. Biol. Chem. PubMed Scopus Google Scholar), and this the tyrosine internalization in the Although ectodomain of APP a role in in cells B. K. B. J. Biol. Chem. PubMed Scopus Google Scholar), for the tyrosine subserving of APP has also been C. A. J. Biol. PubMed Scopus Google It that the tyrosine in APP751 and in APP695) with the protein with homology that with to APP to the in cells P. J. A. PubMed Scopus Google the role of Tyr743 in APP we that Tyr743 in the NPXY is not an APP signal and that cells APP with this mutation normal of Although endocytosis was not in cells with mutations of the NPXY tyrosine also observed normal Aβ secretion from cell lines J. J. Biol. Chem. PubMed Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Although not an signal, reduced the rapid turnover of Although the APP half-life was in Y743A APP was from to glycosylated APP to and suggesting that the was normal for these cell lines. we that Tyr743 in a signal for that cells expressing C-terminal reduced of Aβ secretion J. Biol. Chem. PubMed Google Scholar, K. B. M. Alzheimer's and and Aβ was by cells expressing mutations that reduced APP endocytosis. Y738A, N740A, and cell lines all reduced Aβ secretion by in Aβ secretion was observed for Y709A, G737A, or cell lines that also normal APP endocytosis. the of the for the of Aβ by we Aβ42 secretion by we were to that in parallel with the of Aβ secretion, the Y738A, N740A, and cell lines also secreted that of APP in the to Aβ42 However, J. Biol. Chem. 1996; PubMed Scopus Google Scholar, C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, B. J. A. PubMed Scopus Google Scholar, G. J. M. P. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google Scholar) and cells J. Biol. Chem. 1996; PubMed Scopus Google Scholar, B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar) to Aβ42 in the of the C. A. C. J. Biol. Chem. PubMed Scopus Google Scholar, J. P. J. PubMed Scopus Google B.D. J. Biol. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, M. A. Multhaup G. Masters C.L. Beyreuther K. A. PubMed Scopus Google Scholar, B. Masters C.L. K. 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Selvaraj S, Arnone D, Job D, Stanfield A, Farrow TFD, Nugent AC, Scherk H, Gruber O, Chen X, Sachdev PS, Dickstein DP, Malhi GS, Ha TH, Ha K, Phillips ML, McIntosh AM. Grey matter differences in bipolar disorder: a meta‐analysis of voxel‐based morphometry studies. Bipolar Disord 2012: 14: 135–145. © 2012 The Authors. Journal compilation © 2012 John Wiley & Sons A/S. Objective: Several neuroimaging studies have reported structural brain differences in bipolar disorder using automated methods. While these studies have several advantages over those using region of interest techniques, no study has yet estimated a summary effect size or tested for between‐study heterogeneity. We sought to address this issue using meta‐analytic techniques applied for the first time in bipolar disorder at the level of the individual voxel. Methods: A systematic review identified 16 voxel‐based morphometry (VBM) studies comparing individuals with bipolar disorder with unaffected controls, of which eight were included in the meta‐analysis. In order to take account of heterogeneity, summary effect sizes were computed using a random‐effects model with appropriate correction for multiple testing. Results: Compared with controls, subjects with bipolar disorder had reduced grey matter in a single cluster encompassing the right ventral prefrontal cortex, insula, temporal cortex, and claustrum. Study heterogeneity was widespread throughout the brain; though the significant cluster of grey matter reduction remained once these extraneous voxels had been removed. We found no evidence of publication bias (Eggers p = 0.63). Conclusions: Bipolar disorder is consistently associated with reductions in right prefrontal and temporal lobe grey matter. Reductions elsewhere may be obscured by clinical and methodological heterogeneity.

Abstract Background Although the SARS-CoV-2 viral load detection of respiratory specimen has been widely used for novel coronavirus disease (COVID-19) diagnosis, it is undeniable that serum SARS-CoV-2 nucleic acid (RNAaemia) could be detected in a fraction of the COVID-19 patients. However, it is not clear that if the incidence of RNAaemia could be correlated with the occurrence of cytokine storm or with the specific class of patients. Methods This study enrolled 48 patients with COVID-19 admitted to the General Hospital of Central Theater Command, PLA, a designated hospital in Wuhan, China. The patients were divided into three groups according to the Diagnosis and Treatment of New Coronavirus Pneumonia (version 6) published by the National Health Commission of China. The clinical and laboratory data were collected. The serum viral load detection and serum IL-6 levels were determined. Except for routine statistical analysis, Generalized Linear Models (GLMs) analysis was used to establish a patient status prediction model based on real-time RT-PCR Ct value. Findings The Result showed that cases with RNAaemia were exclusively confirmed in critically ill patients group and appeared to reflect the illness severity. Further more, the inflammatory cytokine IL-6 levels were significantly elevated in critically ill patients, which is almost 10-folds higher than those in other patients. More importantly, the extremely high IL-6 level was closely correlated with the incidence of RNAaemia (R=0.902) and the vital signs of COVID-19 patients (R= −0.682). Interpretation Serum SARS-CoV-2 viral load (RNAaemia) is strongly associated with cytokine storm and can be used to predict the poor prognosis of COVID-19 patients. Moreover, our results strongly suggest that cytokine IL-6 should be considered as a therapeutic target in critically ill patients with excessive inflammatory response.