Guangzhou Institutes of Biomedicine and Health

autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Spatially resolved transcriptomic technologies are promising tools to study complex biological processes such as mammalian embryogenesis. However, the imbalance between resolution, gene capture, and field of view of current methodologies precludes their systematic application to analyze relatively large and three-dimensional mid- and late-gestation embryos. Here, we combined DNA nanoball (DNB)-patterned arrays and in situ RNA capture to create spatial enhanced resolution omics-sequencing (Stereo-seq). We applied Stereo-seq to generate the mouse organogenesis spatiotemporal transcriptomic atlas (MOSTA), which maps with single-cell resolution and high sensitivity the kinetics and directionality of transcriptional variation during mouse organogenesis. We used this information to gain insight into the molecular basis of spatial cell heterogeneity and cell fate specification in developing tissues such as the dorsal midbrain. Our panoramic atlas will facilitate in-depth investigation of longstanding questions concerning normal and abnormal mammalian development.

The genus Coronavirus contains about 25 species of coronaviruses (CoVs), which are important pathogens causing highly prevalent diseases and often severe or fatal in humans and animals. No licensed specific drugs are available to prevent their infection. Different host receptors for cellular entry, poorly conserved structural proteins (antigens), and the high mutation and recombination rates of CoVs pose a significant problem in the development of wide-spectrum anti-CoV drugs and vaccines. CoV main proteases (M(pro)s), which are key enzymes in viral gene expression and replication, were revealed to share a highly conservative substrate-recognition pocket by comparison of four crystal structures and a homology model representing all three genetic clusters of the genus Coronavirus. This conclusion was further supported by enzyme activity assays. Mechanism-based irreversible inhibitors were designed, based on this conserved structural region, and a uniform inhibition mechanism was elucidated from the structures of Mpro-inhibitor complexes from severe acute respiratory syndrome-CoV and porcine transmissible gastroenteritis virus. A structure-assisted optimization program has yielded compounds with fast in vitro inactivation of multiple CoV M(pro)s, potent antiviral activity, and extremely low cellular toxicity in cell-based assays. Further modification could rapidly lead to the discovery of a single agent with clinical potential against existing and possible future emerging CoV-related diseases.

To enhance therapeutic efficacy and reduce adverse effects, practitioners of traditional Chinese medicine (TCM) prescribe a combination of plant species/minerals, called formulae, based on clinical experience. Nearly 100,000 formulae have been recorded, but the working mechanisms of most remain unknown. In trying to address the possible beneficial effects of formulae with current biomedical approaches, we use Realgar-Indigo naturalis formula (RIF), which has been proven to be very effective in treating human acute promyelocytic leukemia (APL) as a model. The main components of RIF are realgar, Indigo naturalis, and Salvia miltiorrhiza, with tetraarsenic tetrasulfide (A), indirubin (I), and tanshinone IIA (T) as major active ingredients, respectively. Here, we report that the ATI combination yields synergy in the treatment of a murine APL model in vivo and in the induction of APL cell differentiation in vitro. ATI causes intensified ubiquitination/degradation of promyelocytic leukemia (PML)-retinoic acid receptor alpha (RARalpha) oncoprotein, stronger reprogramming of myeloid differentiation regulators, and enhanced G(1)/G(0) arrest in APL cells through hitting multiple targets compared with the effects of mono- or biagents. Furthermore, ATI intensifies the expression of Aquaglyceroporin 9 and facilitates the transportation of A into APL cells, which in turn enhances A-mediated PML-RARalpha degradation and therapeutic efficacy. Our data also indicate A as the principal component of the formula, whereas T and I serve as adjuvant ingredients. We therefore suggest that dissecting the mode of action of clinically effective formulae at the molecular, cellular, and organism levels may be a good strategy in exploring the value of traditional medicine.

Mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have been intensely investigated for clinical applications within the last decades. However, the majority of registered clinical trials applying MSC therapy for diverse human diseases have fallen short of expectations, despite the encouraging pre-clinical outcomes in varied animal disease models. This can be attributable to inconsistent criteria for MSCs identity across studies and their inherited heterogeneity. Nowadays, with the emergence of advanced biological techniques and substantial improvements in bio-engineered materials, strategies have been developed to overcome clinical challenges in MSC application. Here in this review, we will discuss the major challenges of MSC therapies in clinical application, the factors impacting the diversity of MSCs, the potential approaches that modify MSC products with the highest therapeutic potential, and finally the usage of MSCs for COVID-19 pandemic disease.

Abstract N 6 -methyladenosine (m 6 A) is the most abundant mRNA modification and is catalyzed by the methyltransferase complex, in which methyltransferase-like 3 (METTL3) is the sole catalytic subunit. Accumulating evidence in recent years reveals that METTL3 plays key roles in a variety of cancer types, either dependent or independent on its m 6 A RNA methyltransferase activity. While the roles of m 6 A modifications in cancer have been extensively reviewed elsewhere, the critical functions of METTL3 in various types of cancer, as well as the potential targeting of METTL3 as cancer treatment, have not yet been highlighted. Here we summarize our current understanding both on the oncogenic and tumor-suppressive functions of METTL3, as well as the underlying molecular mechanisms. The well-documented protein structure of the METTL3/METTL14 heterodimer provides the basis for potential therapeutic targeting, which is also discussed in this review.

The emerging COVID-19 caused by SARS-CoV-2 infection poses severe challenges to global public health. Serum antibody testing is becoming one of the critical methods for the diagnosis of COVID-19 patients. We investigated IgM and IgG responses against SARS-CoV-2 nucleocapsid (N) and spike (S) protein after symptom onset in the intensive care unit (ICU) and non-ICU patients. 130 blood samples from 38 COVID-19 patients were collected. The levels of IgM and IgG specific to N and S protein were detected by ELISA. A series of blood samples were collected along the disease course from the same patient, including 11 ICU patients and 27 non-ICU patients for longitudinal analysis. N and S specific IgM and IgG (N-IgM, N-IgG, S-IgM, S-IgG) in non-ICU patients increased after symptom onset. N-IgM and S-IgM in some non-ICU patients reached a peak in the second week, while N-IgG and S-IgG continued to increase in the third week. The combined detection of N and S specific IgM and IgG could identify up to 75% of SARS-CoV-2 infected patients in the first week. S-IgG was significantly higher in non-ICU patients than in ICU patients in the third week. In contrast, N-IgG was significantly higher in ICU patients than in non-ICU patients. The increase of S-IgG positively correlated with the decrease of C-reactive protein (CRP) in non-ICU patients. N and S specific IgM and IgG increased gradually after symptom onset and can be used for detection of SARS-CoV-2 infection. Analysis of the dynamics of S-IgG may help to predict prognosis.

All-trans retinoic acid (ATRA)/arsenic trioxide (ATO) combination-based therapy has benefitted newly diagnosed acute promyelocytic leukemia (APL) in short-term studies, but the long-term efficacy and safety remained unclear. From April 2001, we have followed 85 patients administrated ATRA/ATO with a median follow-up of 70 months. Eighty patients (94.1%) entered complete remission (CR). Kaplan-Meier estimates of the 5-year event-free survival (EFS) and overall survival (OS) for all patients were 89.2% +/- 3.4% and 91.7% +/- 3.0%, respectively, and the 5-year relapse-free survival (RFS) and OS for patients who achieved CR (n = 80) were 94.8% +/- 2.5% and 97.4% +/- 1.8%, respectively. Upon ATRA/ATO, prognosis was not influenced by initial white blood cell count, distinct PML-RARalpha types, or FLT3 mutations. The toxicity profile was mild and reversible. No secondary carcinoma was observed, and 24 months after the last dose of ATRA/ATO, patients had urine arsenic concentrations well below the safety limit. These results demonstrate the high efficacy and minimal toxicity of ATRA/ATO treatment for newly diagnosed APL in long-term follow-up, suggesting a potential frontline therapy for de novo APL.

Forced expression of selected transcription factors can transform somatic cells into embryonic stem cell (ESC)-like cells, termed induced pluripotent stem cells (iPSCs). There is no consensus regarding the preferred tissue from which to harvest donor cells for reprogramming into iPSCs, and some donor cell types may be more prone than others to accumulation of epigenetic imprints and somatic cell mutations. Here, we present a simple, reproducible, noninvasive method for generating human iPSCs from renal tubular cells present in urine. This procedure eliminates many problems associated with other protocols, and the resulting iPSCs display an excellent ability to differentiate. These data suggest that urine may be a preferred source for generating iPSCs.

Induced pluripotent stem cell (iPS) technology appears to be a general strategy to generate pluripotent stem cells from any given mammalian species. So far, iPS cells have been reported for mouse, human, rat, and monkey. These four species have also established embryonic stem cell (ESC) lines that serve as the gold standard for pluripotency comparisons. Attempts have been made to generate porcine ESC by various means without success. Here we report the successful generation of pluripotent stem cells from fibroblasts isolated from the Tibetan miniature pig using a modified iPS protocol. The resulting iPS cell lines more closely resemble human ESC than cells from other species, have normal karyotype, stain positive for alkaline phosphatase, express high levels of ESC-like markers (Nanog, Rex1, Lin28, and SSEA4), and can differentiate into teratomas composed of the three germ layers. Because porcine physiology closely resembles human, the iPS cells reported here provide an attractive model to study certain human diseases or assess therapeutic applications of iPS in a large animal model. Induced pluripotent stem cell (iPS) technology appears to be a general strategy to generate pluripotent stem cells from any given mammalian species. So far, iPS cells have been reported for mouse, human, rat, and monkey. These four species have also established embryonic stem cell (ESC) lines that serve as the gold standard for pluripotency comparisons. Attempts have been made to generate porcine ESC by various means without success. Here we report the successful generation of pluripotent stem cells from fibroblasts isolated from the Tibetan miniature pig using a modified iPS protocol. The resulting iPS cell lines more closely resemble human ESC than cells from other species, have normal karyotype, stain positive for alkaline phosphatase, express high levels of ESC-like markers (Nanog, Rex1, Lin28, and SSEA4), and can differentiate into teratomas composed of the three germ layers. Because porcine physiology closely resembles human, the iPS cells reported here provide an attractive model to study certain human diseases or assess therapeutic applications of iPS in a large animal model. Induced nuclear reprogramming through induced pluripotent stem cell (iPS) 2The abbreviations used are: iPSinduced pluripotent stem cellESCembryonic stem cellPEFporcine embryonic fibroblastsAPalkaline phosphataseTRITCtetramethylrhodamine isothiocyanateRTreverse transcriptase. 2The abbreviations used are: iPSinduced pluripotent stem cellESCembryonic stem cellPEFporcine embryonic fibroblastsAPalkaline phosphataseTRITCtetramethylrhodamine isothiocyanateRTreverse transcriptase. technology is an amazing achievement full of challenge to the intellect and important practical implications (1Tweedell K.S. Curr. Stem Cell Res. Ther. 2008; 3: 151-162Crossref PubMed Scopus (24) Google Scholar, 2Pei D. Cell Res. 2008; 18: 221-223Crossref PubMed Scopus (16) Google Scholar). Overexpression of exogenous factors that are highly enriched in embryonic stem cell (ESC) can rearrange the genetic program of different cell types, including somatic and adult stem cells, and induce a long lasting ESC-like pluripotent state (3Takahashi K. Yamanaka S. Cell. 2006; 126: 663-676Abstract Full Text Full Text PDF PubMed Scopus (18482) Google Scholar, 4Okita K. Ichisaka T. Yamanaka S. Nature. 2007; 448: 313-317Crossref PubMed Scopus (3498) Google Scholar, 5Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14662) Google Scholar, 6Yu J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8046) Google Scholar, 7Wernig M. Meissner A. Foreman R. Brambrink T. Ku M. Hochedlinger K. Bernstein B.E. Jaenisch R. Nature. 2007; 448: 318-324Crossref PubMed Scopus (2204) Google Scholar). The repercussions of iPS technology are vast: it provides a way to create patient-specific stem cells that bypasses ethical and technical issues surrounding human ESC derivation and somatic cell nuclear transfer (8Thomson J.A. Kalishman J. Golos T.G. Durning M. Harris C.P. Becker R.A. Hearn J.P. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 7844-7848Crossref PubMed Scopus (909) Google Scholar, 9Wilmut I. Schnieke A.E. McWhir J. Kind A.J. Campbell K.H. Nature. 1997; 385: 810-813Crossref PubMed Scopus (4010) Google Scholar), a state of the art model for studying genetic diseases in vitro (10Park I.H. Arora N. Huo H. Maherali N. Ahfeldt T. Shimamura A. Lensch M.W. Cowan C. Hochedlinger K. Daley G.Q. Cell. 2008; 134: 877-886Abstract Full Text Full Text PDF PubMed Scopus (1790) Google Scholar, 11Ebert A.D. Yu J. Rose Jr., F.F. Mattis V.B. Lorson C.L. Thomson J.A. Svendsen C.N. Nature. 2009; 457: 277-280Crossref PubMed Scopus (1170) Google Scholar), and an incredible backwards route that can crystallize our current understanding of developmental and stem cell biology. Many questions, especially mechanistic, remain unanswered, but the current rhythm of research may bring iPS to clinical application sooner than expected. However, before jumping onto such extraordinary endeavor, safety must be scrupulously tested in an animal model close enough to humans. Nowadays that iPS technology is expanding, with improved delivery systems, chemical additions, new tissue culture conditions, and multiple cell sources being reported regularly, such animal model is essential to set up quality standards (12Kaji K. Norrby K. Paca A. Mileikovsky M. Mohseni P. Woltjen K. Nature. 2009; 458: 771-775Crossref PubMed Scopus (1035) Google Scholar, 13Woltjen K. Michael I.P. Mohseni P. Desai R. Mileikovsky M. Hämäläinen R. Cowling R. Wang W. Liu P. Gertsenstein M. Kaji K. Sung H.K. Nagy A. Nature. 2009; 458: 766-770Crossref PubMed Scopus (1432) Google Scholar, 14Okita K. Nakagawa M. Hyenjong H. Ichisaka T. Yamanaka S. Science. 2008; 322: 949-953Crossref PubMed Scopus (1588) Google Scholar, 15Stadtfeld M. Nagaya M. Utikal J. Weir G. Hochedlinger K. Science. 2008; 322: 945-949Crossref PubMed Scopus (1321) Google Scholar, 16Aoi T. Yae K. Nakagawa M. Ichisaka T. Okita K. Takahashi K. Chiba T. Yamanaka S. Science. 2008; 321: 699-702Crossref PubMed Scopus (859) Google Scholar, 17Hanna J. Markoulaki S. Schorderet P. Carey B.W. Beard C. Wernig M. Creyghton M.P. Steine E.J. Cassady J.P. Foreman R. Lengner C.J. Dausman J.A. Jaenisch R. Cell. 2008; 133: 250-264Abstract Full Text Full Text PDF PubMed Scopus (674) Google Scholar, C. S. Cell Stem Cell. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). and as the to reprogramming and but and are for safety in humans. are an but in ethical remain to and are to to a of have of and physiology is to up as the attractive model for from is used to pig and have and to human for V. Stem Cell 2008; PubMed Scopus Google Scholar). have also for of a in of more genetic D. M. S. J. J.L. Liu C.N. A. E.J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). of ESC from or animal species from mouse, human, and more the rat, of A. Stem Cell 2008; PubMed Scopus Google Scholar, I. A. J. 2007; PubMed Scopus Google Scholar). porcine genetic can be through and somatic cell nuclear transfer D. J.L. M. A. A. C.N. Science. PubMed Scopus Google Scholar). porcine embryonic fibroblasts the cell for somatic cell nuclear have a that D. C. A. M. P. J. P. Stem 2009; PubMed Scopus (24) Google Scholar). of pig iPS cell lines provide an of for a different from porcine iPS may as the of the miniature pig as a for of the pig is and and Liu Yu M. K. 2006; PubMed Scopus Google Scholar). of and the standard to induce and been used in mouse, human, and cells K. Ichisaka T. Yamanaka S. Nature. 2007; 448: 313-317Crossref PubMed Scopus (3498) Google Scholar, 5Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14662) Google Scholar, H. J. P. P. H. W. J. Liu M. K. T. D. G. W. M. H. Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar, W. W. S. J. T. A. H. S. Cell Stem Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). factors can human cells P. Yu J. G. J. Stem 2008; PubMed Scopus Google Scholar). and human factors by means of and used as a with close to culture for standard ESC and modified high with of ESC as and but with and a and of the for the cells in a set as is the in and we it the with or human iPS generation in cells onto with human ESC-like and with more the in and of the factors cell also and to more standard with human ESC-like in the and of in the and of in the human, the and positive for These cell lines be without but or without induced and the a high with of human and H. J. P. P. H. W. J. Liu M. K. T. D. G. W. M. H. Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar), and positive of the into the of tested iPS cell lines an of high or in and high in iPS of of pluripotency porcine iPS positive for the human and and factors and in or iPS cell lines resulting from or human or in high of with that the and also of and for the also been by in human and iPS cell lines J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8046) Google Scholar, J. C. S. J. J. H. N. H. Cell Stem Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). can the of iPS cell lines into full iPS J. Ku M. Wernig M. Schorderet P. Bernstein B.E. Jaenisch R. Meissner A. Nature. 2008; PubMed Scopus Google Scholar, J. J. J. A. 2008; PubMed Scopus Google Scholar). of pig iPS cell lines with a of any in cell or ESC markers of and and but levels of the tested ESC markers pig iPS cell lines into the of in teratomas from the three germ and and of pig iPS cell lines that pluripotent with of and have been of isolated porcine cells into V. Stem Cell 2008; PubMed Scopus Google Scholar). the of our study we pig iPS cell lines into from and into The of such is our of using pig ESC that been for to of pig iPS and the of from pig ESC been reported V. Stem Cell 2008; PubMed Scopus Google of porcine iPS cell with ESC-like with a are pig iPS is the and positive for is in with of exogenous or human factors into the of pig iPS to and for the high in iPS with with iPS and M. H. of porcine iPS cell of the ESC are the and for iPS that and serve as an are of iPS cell the and are also and used as used as of human factors and iPS with of ESC markers iPS and with that the of the in iPS cell lines with from cells used as for the M. H. iPS cell lines are in into the three germ layers. from the are a with are the and is for and are in and a in and tissue is in an of in different human iPS cell lines with that the iPS cell lines to different H. we a for reprogramming into iPS cells, and provide for The of of for the in our cell lines are may have an the of our cell lines to differentiate into different and may the long for The of different cell other than fibroblasts a iPS generation D. K. Wang H. W. Wang T. W. J. G. M.A. S. D. J. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, V. G. P. K. D. M. D. J. K. C. C. M. H. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar, S. Utikal J. K. Jaenisch R. Hochedlinger K. Stem 2008; PubMed Scopus Google Scholar). more of to iPS different porcine be important and full by the of the from an cell that the animal be for of exogenous factors also of the exogenous and we are up for porcine iPS D. Beard C. G. A. M. Jaenisch R. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). of as to be in pig may from established and human and in in are the may be The can be or other for the Tibetan in of have for of in a and have that to iPS of cells from other pig be important as are using in iPS cell lines be the long of iPS generation is an it is in The the in of can be more than the and in the of an model for porcine iPS research is and Induced nuclear reprogramming through induced pluripotent stem cell (iPS) 2The abbreviations used are: iPSinduced pluripotent stem cellESCembryonic stem cellPEFporcine embryonic fibroblastsAPalkaline phosphataseTRITCtetramethylrhodamine isothiocyanateRTreverse transcriptase. 2The abbreviations used are: iPSinduced pluripotent stem cellESCembryonic stem cellPEFporcine embryonic fibroblastsAPalkaline phosphataseTRITCtetramethylrhodamine isothiocyanateRTreverse transcriptase. technology is an amazing achievement full of challenge to the intellect and important practical implications (1Tweedell K.S. Curr. Stem Cell Res. Ther. 2008; 3: 151-162Crossref PubMed Scopus (24) Google Scholar, 2Pei D. Cell Res. 2008; 18: 221-223Crossref PubMed Scopus (16) Google Scholar). Overexpression of exogenous factors that are highly enriched in embryonic stem cell (ESC) can rearrange the genetic program of different cell types, including somatic and adult stem cells, and induce a long lasting ESC-like pluripotent state (3Takahashi K. Yamanaka S. Cell. 2006; 126: 663-676Abstract Full Text Full Text PDF PubMed Scopus (18482) Google Scholar, 4Okita K. Ichisaka T. Yamanaka S. Nature. 2007; 448: 313-317Crossref PubMed Scopus (3498) Google Scholar, 5Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14662) Google Scholar, 6Yu J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8046) Google Scholar, 7Wernig M. Meissner A. Foreman R. Brambrink T. Ku M. Hochedlinger K. Bernstein B.E. Jaenisch R. Nature. 2007; 448: 318-324Crossref PubMed Scopus (2204) Google Scholar). The repercussions of iPS technology are vast: it provides a way to create patient-specific stem cells that bypasses ethical and technical issues surrounding human ESC derivation and somatic cell nuclear transfer (8Thomson J.A. Kalishman J. Golos T.G. Durning M. Harris C.P. Becker R.A. Hearn J.P. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 7844-7848Crossref PubMed Scopus (909) Google Scholar, 9Wilmut I. Schnieke A.E. McWhir J. Kind A.J. Campbell K.H. Nature. 1997; 385: 810-813Crossref PubMed Scopus (4010) Google Scholar), a state of the art model for studying genetic diseases in vitro (10Park I.H. Arora N. Huo H. Maherali N. Ahfeldt T. Shimamura A. Lensch M.W. Cowan C. Hochedlinger K. Daley G.Q. Cell. 2008; 134: 877-886Abstract Full Text Full Text PDF PubMed Scopus (1790) Google Scholar, 11Ebert A.D. Yu J. Rose Jr., F.F. Mattis V.B. Lorson C.L. Thomson J.A. Svendsen C.N. Nature. 2009; 457: 277-280Crossref PubMed Scopus (1170) Google Scholar), and an incredible backwards route that can crystallize our current understanding of developmental and stem cell biology. Many questions, especially mechanistic, remain unanswered, but the current rhythm of research may bring iPS to clinical application sooner than expected. However, before jumping onto such extraordinary endeavor, safety must be scrupulously tested in an animal model close enough to humans. Nowadays that iPS technology is expanding, with improved delivery systems, chemical additions, new tissue culture conditions, and multiple cell sources being reported regularly, such animal model is essential to set up quality standards (12Kaji K. Norrby K. Paca A. Mileikovsky M. Mohseni P. Woltjen K. Nature. 2009; 458: 771-775Crossref PubMed Scopus (1035) Google Scholar, 13Woltjen K. Michael I.P. Mohseni P. Desai R. Mileikovsky M. Hämäläinen R. Cowling R. Wang W. Liu P. Gertsenstein M. Kaji K. Sung H.K. Nagy A. Nature. 2009; 458: 766-770Crossref PubMed Scopus (1432) Google Scholar, 14Okita K. Nakagawa M. Hyenjong H. Ichisaka T. Yamanaka S. Science. 2008; 322: 949-953Crossref PubMed Scopus (1588) Google Scholar, 15Stadtfeld M. Nagaya M. Utikal J. Weir G. Hochedlinger K. Science. 2008; 322: 945-949Crossref PubMed Scopus (1321) Google Scholar, 16Aoi T. Yae K. Nakagawa M. Ichisaka T. Okita K. Takahashi K. Chiba T. Yamanaka S. Science. 2008; 321: 699-702Crossref PubMed Scopus (859) Google Scholar, 17Hanna J. Markoulaki S. Schorderet P. Carey B.W. Beard C. Wernig M. Creyghton M.P. Steine E.J. Cassady J.P. Foreman R. Lengner C.J. Dausman J.A. Jaenisch R. Cell. 2008; 133: 250-264Abstract Full Text Full Text PDF PubMed Scopus (674) Google Scholar, C. S. Cell Stem Cell. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar). and as the to reprogramming and but and are for safety in humans. are an but in ethical remain to and are to to a of have of and physiology is to up as the attractive model for from is used to pig and have and to human for V. Stem Cell 2008; PubMed Scopus Google Scholar). have also for of a in of more genetic D. M. S. J. J.L. Liu C.N. A. E.J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). of ESC from or animal species from mouse, human, and more the rat, of A. Stem Cell 2008; PubMed Scopus Google Scholar, I. A. J. 2007; PubMed Scopus Google Scholar). porcine genetic can be through and somatic cell nuclear transfer D. J.L. M. A. A. C.N. Science. PubMed Scopus Google Scholar). porcine embryonic fibroblasts the cell for somatic cell nuclear have a that D. C. A. M. P. J. P. Stem 2009; PubMed Scopus (24) Google Scholar). of pig iPS cell lines provide an of for a different from porcine iPS may as the of induced pluripotent stem cell embryonic stem cell porcine embryonic fibroblasts alkaline transcriptase. induced pluripotent stem cell embryonic stem cell porcine embryonic fibroblasts alkaline transcriptase. the miniature pig as a for of the pig is and and Liu Yu M. K. 2006; PubMed Scopus Google Scholar). of and the standard to induce and been used in mouse, human, and cells K. Ichisaka T. Yamanaka S. Nature. 2007; 448: 313-317Crossref PubMed Scopus (3498) Google Scholar, 5Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14662) Google Scholar, H. J. P. P. H. W. J. Liu M. K. T. D. G. W. M. H. Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar, W. W. S. J. T. A. H. S. Cell Stem Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). factors can human cells P. Yu J. G. J. Stem 2008; PubMed Scopus Google Scholar). and human factors by means of and used as a with close to culture for standard ESC and modified high with of ESC as and but with and a and of the for the cells in a set as is the in and we it the with or human iPS generation in cells onto with human ESC-like and with more the in and of the factors cell also and to more standard with human ESC-like in the and of in the and of in the human, the and positive for These cell lines be without but or without induced and the a high with of human and H. J. P. P. H. W. J. Liu M. K. T. D. G. W. M. H. Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar), and positive of the into the of tested iPS cell lines an of high or in and high in iPS of of pluripotency porcine iPS positive for the human and and factors and in or iPS cell lines resulting from or human or in high of with that the and also of and for the also been by in human and iPS cell lines J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8046) Google Scholar, J. C. S. J. J. H. N. H. Cell Stem Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). can the of iPS cell lines into full iPS J. Ku M. Wernig M. Schorderet P. Bernstein B.E. Jaenisch R. Meissner A. Nature. 2008; PubMed Scopus Google Scholar, J. J. J. A. 2008; PubMed Scopus Google Scholar). of pig iPS cell lines with a of any in cell or ESC markers of and and but levels of the tested ESC markers pig iPS cell lines into the of in teratomas from the three germ and and of pig iPS cell lines that pluripotent with of and have been of isolated porcine cells into V. Stem Cell 2008; PubMed Scopus Google Scholar). the of our study we pig iPS cell lines into from and into The of such is our of using pig ESC that been for to of pig iPS and the of from pig ESC been reported V. Stem Cell 2008; PubMed Scopus Google of porcine iPS cell of the ESC are the and for iPS that and serve as an are of iPS cell the and are also and used as used as of human factors and iPS with of ESC markers iPS and with that the of the in iPS cell lines with from cells used as for the M. H. iPS cell lines are in into the three germ layers. from the are a with are the and is for and are in and a in and tissue is in an of in different human iPS cell lines with that the iPS cell lines to different H. we a for reprogramming into iPS cells, and provide for The of of for the in our cell lines are may have an the of our cell lines to differentiate into different and may the long for The of different cell other than fibroblasts a iPS generation D. K. Wang H. W. Wang T. W. J. G. M.A. S. D. J. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, V. G. P. K. D. M. D. J. K. C. C. M. H. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar, S. Utikal J. K. Jaenisch R. Hochedlinger K. Stem 2008; PubMed Scopus Google Scholar). more of to iPS different porcine be important and full by the of the from an cell that the animal be for of exogenous factors also of the exogenous and we are up for porcine iPS D. Beard C. G. A. M. Jaenisch R. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). of as to be in pig may from established and human and in in are the may be The can be or other for the Tibetan in of have for of in a and have that to iPS of cells from other pig be important as are using in iPS cell lines be the long of iPS generation is an it is in The the in of can be more than the and in the of an model for porcine iPS research is and the miniature pig as a for of the pig is and and Liu Yu M. K. 2006; PubMed Scopus Google Scholar). of and the standard to induce and been used in mouse, human, and cells K. Ichisaka T. Yamanaka S. Nature. 2007; 448: 313-317Crossref PubMed Scopus (3498) Google Scholar, 5Takahashi K. Tanabe K. Ohnuki M. Narita M. Ichisaka T. Tomoda K. Yamanaka S. Cell. 2007; 131: 861-872Abstract Full Text Full Text PDF PubMed Scopus (14662) Google Scholar, H. J. P. P. H. W. J. Liu M. K. T. D. G. W. M. H. Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar, W. 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Cell Stem Cell. 2008; 3: Full Text Full Text PDF PubMed Scopus Google Scholar), and positive of the into the of tested iPS cell lines an of high or in and high in iPS of of pluripotency porcine iPS positive for the human and and factors and in or iPS cell lines resulting from or human or in high of with that the and also of and for the also been by in human and iPS cell lines J. Vodyanik M.A. Smuga-Otto K. Antosiewicz-Bourget J. Frane J.L. Tian S. Nie J. Jonsdottir G.A. Ruotti V. Stewart R. Slukvin I.I. Thomson J.A. Science. 2007; 318: 1917-1920Crossref PubMed Scopus (8046) Google Scholar, J. C. S. J. J. H. N. H. Cell Stem Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). can the of iPS cell lines into full iPS J. Ku M. Wernig M. Schorderet P. Bernstein B.E. Jaenisch R. Meissner A. Nature. 2008; PubMed Scopus Google Scholar, J. J. J. A. 2008; PubMed Scopus Google Scholar). of pig iPS cell lines with a of any in cell or ESC markers of and and but levels of the tested ESC markers pig iPS cell lines into the of in teratomas from the three germ and and of pig iPS cell lines that pluripotent with of and have been of isolated porcine cells into V. Stem Cell 2008; PubMed Scopus Google Scholar). the of our study we pig iPS cell lines into from and into The of such is our of using pig ESC that been for to of pig iPS and the of from pig ESC been reported V. Stem Cell 2008; PubMed Scopus Google Scholar). we a for reprogramming into iPS cells, and provide for The of of for the in our cell lines are may have an the of our cell lines to differentiate into different and may the long for The of different cell other than fibroblasts a iPS generation D. K. Wang H. W. Wang T. W. J. G. M.A. S. D. J. 2008; Full Text Full Text PDF PubMed Scopus Google Scholar, V. G. P. K. D. M. D. J. K. C. C. M. H. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar, S. Utikal J. K. Jaenisch R. Hochedlinger K. Stem 2008; PubMed Scopus Google Scholar). more of to iPS different porcine be important and full by the of the from an cell that the animal be for of exogenous factors also of the exogenous and we are up for porcine iPS D. Beard C. G. A. M. Jaenisch R. Cell. 2009; Full Text Full Text PDF PubMed Scopus Google Scholar). of as to be in pig may from established and human and in in are the may be The can be or other for the Tibetan in of have for of in a and have that to iPS of cells from other pig be important as are using in iPS cell lines be the long of iPS generation is an it is in The the in of can be more than the and in the of an model for porcine iPS research is and of the for for and the of

SARS-CoV-2 caused a major outbreak of severe pneumonia (COVID-19) in humans. Viral RNA was detected in multiple organs in COVID-19 patients. However, infectious SARS-CoV-2 was only isolated from respiratory specimens. Here, infectious SARS-CoV-2 was successfully isolated from urine of a COVID-19 patient. The virus isolated could infect new susceptible cells and was recognized by its' own patient sera. Appropriate precautions should be taken to avoid transmission from urine.

Abstract Tissue regeneration requires coordination between resident stem cells and local niche cells 1,2 . Here we identify that senescent cells are integral components of the skeletal muscle regenerative niche that repress regeneration at all stages of life. The technical limitation of senescent-cell scarcity 3 was overcome by combining single-cell transcriptomics and a senescent-cell enrichment sorting protocol. We identified and isolated different senescent cell types from damaged muscles of young and old mice. Deeper transcriptome, chromatin and pathway analyses revealed conservation of cell identity traits as well as two universal senescence hallmarks (inflammation and fibrosis) across cell type, regeneration time and ageing. Senescent cells create an aged-like inflamed niche that mirrors inflammation associated with ageing (inflammageing 4 ) and arrests stem cell proliferation and regeneration. Reducing the burden of senescent cells, or reducing their inflammatory secretome through CD36 neutralization, accelerates regeneration in young and old mice. By contrast, transplantation of senescent cells delays regeneration. Our results provide a technique for isolating in vivo senescent cells, define a senescence blueprint for muscle, and uncover unproductive functional interactions between senescent cells and stem cells in regenerative niches that can be overcome. As senescent cells also accumulate in human muscles, our findings open potential paths for improving muscle repair throughout life.

A novel and efficient synthesis of pyrido[1,2-a]benzimidazoles through direct intramolecular aromatic C-H amination of N-aryl-2-aminopyridines has been developed. The reaction, cocatalyzed by Cu(OAc)(2) and Fe(NO(3))(3)·9H(2)O, is carried out in DMF under a dioxygen atmosphere. Diversified pyrido[1,2-a]benzimidazoles containing various substitution patterns are obtained in moderate to excellent yields by using this procedure. The results of mechanistic studies suggest that a Cu(III)-catalyzed electrophilic aromatic substitution (S(E)Ar) pathway is operating in this process. The unique role of iron(III) is believed to lie in its ability to facilitate formation of the more electrophilic Cu(III) species. In the absence of iron(III), a much less efficient and reversible Cu(II)-mediated S(E)Ar process takes place.

Growing interest in extracellular vesicles (EVs, including exosomes and microvesicles) as therapeutic entities, particularly in stem cell-related approaches, has underlined the need for standardization and coordination of development efforts. Members of the International Society for Extracellular Vesicles and the Society for Clinical Research and Translation of Extracellular Vesicles Singapore convened a Workshop on this topic to discuss the opportunities and challenges associated with development of EV-based therapeutics at the preclinical and clinical levels. This review outlines topic-specific action items that, if addressed, will enhance the development of best-practice models for EV therapies. Stem Cells Translational Medicine 2017;6:1730-1739.

Increasing evidence suggests circular RNAs (circRNAs) exert critical functions in tumor progression via sponging miRNAs (microRNAs). However, the role of circRNAs in breast cancer remains unclear. Here we systematically analyzed the circular RNAs in breast cancer based on their characteristic in sponging disease-specific miRNAs and identified hsa_circ_001783 as a top ranked circRNA in our computation and verified its high expression in both breast cancer cells and cancer tissue. A higher level of hsa_circ_001783 was significantly correlated with heavier tumor burden and poorer prognosis of patients with breast cancer. Knockdown of this circRNA remarkably inhibited the proliferation and invasion of breast cancer cells. Importantly, hsa_circ_001783 promoted progression of breast cancer cells via sponging miR-200c-3p. Taken together, hsa_circ_001783 may serve as a novel prognostic and therapeutic target for breast cancer.

Classic embryological experiments have established that the early mouse embryo develops via sequential lineage bifurcations. The first segregated lineage is the trophectoderm, essential for blastocyst formation. Mouse naive epiblast and derivative embryonic stem cells are restricted accordingly from producing trophectoderm. Here we show, in contrast, that human naive embryonic stem cells readily make blastocyst trophectoderm and descendant trophoblast cell types. Trophectoderm was induced rapidly and efficiently by inhibition of ERK/mitogen-activated protein kinase (MAPK) and Nodal signaling. Transcriptome comparison with the human embryo substantiated direct formation of trophectoderm with subsequent differentiation into syncytiotrophoblast, cytotrophoblast, and downstream trophoblast stem cells. During pluripotency progression lineage potential switches from trophectoderm to amnion. Live-cell tracking revealed that epiblast cells in the human blastocyst are also able to produce trophectoderm. Thus, the paradigm of developmental specification coupled to lineage restriction does not apply to humans. Instead, epiblast plasticity and the potential for blastocyst regeneration are retained until implantation.

A direct synthesis of carbaldehydes through intramolecular dehydrogenative aminooxygenation has been developed. The process uses a catalytic amount of copper(II) in DMF or DMA under oxygen and does not require additional oxidants (see scheme). Mechanistic studies suggest that the carbonyl oxygen atom of the aldehyde is derived from oxygen through a copper-mediated oxygen activation process via a peroxy–copper(III) intermediate.

Not only symmetrical, but also unsymmetrical α,α-diaryl allylic alcohols are employed as substrates in the title reaction. A number of arenes and even heteroarenes underwent radical 1,2-aryl migration (“neophyl rearrangement”) to produce α-aryl β-trifluoromethyl ketones. The preferential migration of electron-deficient aryl groups over electron-rich ones in unsymmetrical substrates supports the radical mechanism, which was further confirmed by DFT calculations. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Adiponectin protects the vascular system partly through stimulation of endothelial nitric oxide (NO) production and endothelium-dependent vasodilation. The current study investigated the role of two recently identified adiponectin receptors, AdipoR1 and -R2, and their downstream effectors in mediating the endothelium actions of adiponectin. In human umbilical vein endothelial cells, adiponectin-induced phosphorylation of endothelial NO synthase (eNOS) at Ser(1177) and NO production were abrogated when expression of AdipoR1 and -R2 were simultaneously suppressed. Proteomic analysis demonstrated that the cytoplasmic tails of both AdipoR1 and -R2 interacted with APPL1, an adaptor protein that contains a PH (pleckstrin homology) domain, a PTB (phosphotyrosine-binding) domain, and a Leucine zipper motif. Suppression of APPL1 expression by RNA interference significantly attenuated adiponectin-induced phosphorylation of AMP-activated protein kinase (AMPK) at Thr(172) and eNOS at Ser(1177), and the complex formation between eNOS and heat shock protein 90, resulting in a marked reduction of NO production. Adenovirus-mediated overexpression of a constitutively active version of AMPK reversed these changes. In db/db diabetic mice, both APPL1 expression and adiponectin-induced vasodilation were significantly decreased compared with their lean littermates. Taken together, these results suggest that APPL1 acts as a common downstream effector of AdipoR1 and -R2, mediating adiponectin-evoked endothelial NO production and endothelium-dependent vasodilation.

Importance: Axillary lymph node metastasis (ALNM) status, typically estimated using an invasive procedure with a high false-negative rate, strongly affects the prognosis of recurrence in breast cancer. However, preoperative noninvasive tools to accurately predict ALNM status and disease-free survival (DFS) are lacking. Objective: To develop and validate dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) radiomic signatures for preoperative identification of ALNM and to assess individual DFS in patients with early-stage breast cancer. Design, Setting, and Participants: This retrospective prognostic study included patients with histologically confirmed early-stage breast cancer diagnosed at 4 hospitals in China from July 3, 2007, to September 21, 2019, randomly divided (7:3) into development and vaidation cohorts. All patients underwent preoperative MRI scans, were treated with surgery and sentinel lymph node biopsy or ALN dissection, and were pathologically examined to determine the ALNM status. Data analysis was conducted from February 15, 2019, to March 20, 2020. Exposure: Clinical and DCE-MRI radiomic signatures. Main Outcomes and Measures: The primary end points were ALNM and DFS. Results: This study included 1214 women (median [IQR] age, 47 [42-55] years), split into development (849 [69.9%]) and validation (365 [30.1%]) cohorts. The radiomic signature identified ALNM in the development and validation cohorts with areas under the curve (AUCs) of 0.88 and 0.85, respectively, and the clinical-radiomic nomogram accurately predicted ALNM in the development and validation cohorts (AUC, 0.92 and 0.90, respectively) based on a least absolute shrinkage and selection operator (LASSO)-logistic regression model. The radiomic signature predicted 3-year DFS in the development and validation cohorts (AUC, 0.81 and 0.73, respectively), and the clinical-radiomic nomogram could discriminate high-risk from low-risk patients in the development cohort (hazard ratio [HR], 0.04; 95% CI, 0.01-0.11; P < .001) and the validation cohort (HR, 0.04; 95% CI, 0.004-0.32; P < .001) based on a random forest-Cox regression model. The clinical-radiomic nomogram was associated with 3-year DFS in the development and validation cohorts (AUC, 0.89 and 0.90, respectively). The decision curve analysis demonstrated that the clinical-radiomic nomogram displayed better clinical predictive usefulness than the clinical or radiomic signature alone. Conclusions and Relevance: This study described the application of MRI-based machine learning in patients with breast cancer, presenting novel individualized clinical decision nomograms that could be used to predict ALNM status and DFS. The clinical-radiomic nomograms were useful in clinical decision-making associated with personalized selection of surgical interventions and therapeutic regimens for patients with early-stage breast cancer.