Third Affiliated Hospital of Zhengzhou University

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.

BackgroundOn January 20, 2020, a new coronavirus epidemic with human-to-human transmission was officially declared by the Chinese government, which caused significant public panic in China. In light of the coronavirus disease 2019 outbreak, pregnant women may be particularly vulnerable and in special need for preventive mental health strategies. Thus far, no reports exist to investigate the mental health response of pregnant women to the coronavirus disease 2019 outbreak.ObjectiveThis study aimed to examine the impact of coronavirus disease 2019 outbreak on the prevalence of depressive and anxiety symptoms and the corresponding risk factors among pregnant women across China.Study DesignA multicenter, cross-sectional study was initiated in early December 2019 to identify mental health concerns in pregnancy using the Edinburgh Postnatal Depression Scale. This study provided a unique opportunity to compare the mental status of pregnant women before and after the declaration of the coronavirus disease 2019 epidemic. A total of 4124 pregnant women during their third trimester from 25 hospitals in 10 provinces across China were examined in this cross-sectional study from January 1, 2020, to February 9, 2020. Of these women, 1285 were assessed after January 20, 2020, when the coronavirus epidemic was publicly declared and 2839 were assessed before this pivotal time point. The internationally recommended Edinburgh Postnatal Depression Scale was used to assess maternal depression and anxiety symptoms. Prevalence rates and risk factors were compared between the pre- and poststudy groups.ResultsPregnant women assessed after the declaration of coronavirus disease 2019 epidemic had significantly higher rates of depressive symptoms (26.0% vs 29.6%, P=.02) than women assessed before the epidemic declaration. These women were also more likely to have thoughts of self-harm (P=.005). The depressive rates were positively associated with the number of newly confirmed cases of coronavirus disease 2019 (P=.003), suspected infections (P=.004), and deaths per day (P=.001). Pregnant women who were underweight before pregnancy, primiparous, younger than 35 years, employed full time, in middle income category, and had appropriate living space were at increased risk for developing depressive and anxiety symptoms during the outbreak.ConclusionMajor life-threatening public health events such as the coronavirus disease 2019 outbreak may increase the risk for mental illness among pregnant women, including thoughts of self-harm. Strategies targeting maternal stress and isolation such as effective risk communication and the provision of psychological first aid may be particularly useful to prevent negative outcomes for women and their fetuses. On January 20, 2020, a new coronavirus epidemic with human-to-human transmission was officially declared by the Chinese government, which caused significant public panic in China. In light of the coronavirus disease 2019 outbreak, pregnant women may be particularly vulnerable and in special need for preventive mental health strategies. Thus far, no reports exist to investigate the mental health response of pregnant women to the coronavirus disease 2019 outbreak. This study aimed to examine the impact of coronavirus disease 2019 outbreak on the prevalence of depressive and anxiety symptoms and the corresponding risk factors among pregnant women across China. A multicenter, cross-sectional study was initiated in early December 2019 to identify mental health concerns in pregnancy using the Edinburgh Postnatal Depression Scale. This study provided a unique opportunity to compare the mental status of pregnant women before and after the declaration of the coronavirus disease 2019 epidemic. A total of 4124 pregnant women during their third trimester from 25 hospitals in 10 provinces across China were examined in this cross-sectional study from January 1, 2020, to February 9, 2020. Of these women, 1285 were assessed after January 20, 2020, when the coronavirus epidemic was publicly declared and 2839 were assessed before this pivotal time point. The internationally recommended Edinburgh Postnatal Depression Scale was used to assess maternal depression and anxiety symptoms. Prevalence rates and risk factors were compared between the pre- and poststudy groups. Pregnant women assessed after the declaration of coronavirus disease 2019 epidemic had significantly higher rates of depressive symptoms (26.0% vs 29.6%, P=.02) than women assessed before the epidemic declaration. These women were also more likely to have thoughts of self-harm (P=.005). The depressive rates were positively associated with the number of newly confirmed cases of coronavirus disease 2019 (P=.003), suspected infections (P=.004), and deaths per day (P=.001). Pregnant women who were underweight before pregnancy, primiparous, younger than 35 years, employed full time, in middle income category, and had appropriate living space were at increased risk for developing depressive and anxiety symptoms during the outbreak. Major life-threatening public health events such as the coronavirus disease 2019 outbreak may increase the risk for mental illness among pregnant women, including thoughts of self-harm. Strategies targeting maternal stress and isolation such as effective risk communication and the provision of psychological first aid may be particularly useful to prevent negative outcomes for women and their fetuses.

Objective The aim of this study was to further clarify clinical characteristics and predict mortality risk among patients with viral pneumonia. Methods A total of 528 patients with viral pneumonia at RuiJin hospital in Shanghai from May 2015 to May 2019 were recruited. Multiplex real-time RT-PCR was used to detect respiratory viruses. Demographic information, comorbidities, routine laboratory examinations, immunological indexes, etiological detections, radiological images and treatment were collected on admission. Results 76 (14.4%) patients died within 90 days in hospital. A predictive MuLBSTA score was calculated on the basis of a multivariate logistic regression model in order to predict mortality with a weighted score that included multilobular infiltrates (OR=5.20, 95%CI 1.41-12.52, p=0.010; 5 points), lymphocyte≤0.8*109/L (OR=4.53, 95%CI 2.55-8.05, p<0.001; 4 points), bacterial coinfection (OR=3.71, 95%CI 2.11-6.51, p<0.001; 4 points), acute-smoker (OR=3.19, 95% CI 1.34-6.26, p=0.001; 3 points), quit-smoker (OR=2.18, 95% CI 0.99-4.82, p=0.054; 2 points), hypertension (OR=2.39, 95% CI 1.55-4.26, p=0.003; 2 points) and age≥60 years (OR=2.14, 95% CI 1.04-4.39, p=0.038; 2 points). 12 points was used as a cut-off value for mortality risk stratification. This model showed sensitivity of 0.776, specificity of 0.778 and a better predictive ability than CURB-65 (AUROC = 0.773 vs 0.717, p<0.001). Conclusions Here, we designed an easy-to-use clinically predictive tool for assessing 90-day mortality risk of viral pneumonia. It can accurately stratify hospitalized patients with viral pneumonia into relevant risk categories and could provide guidance to make further clinical decisions.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder that is characterized by impairments in social interactions and communication, restricted interests and repetitive behaviors. Several studies report a high prevalence of gastrointestinal (GI) symptoms in autistic individuals. Cumulative evidence reveals that the gut microbiota and its metabolites (especially short-chain fatty acids, SCFAs) play an important role in GI disorders and the pathogenesis of ASD. However, the composition of the gut microbiota and its association with fecal SCFAs and GI symptoms of autistic children remain largely unknown. In the present study, we sequenced the bacterial 16S rRNA gene, detected fecal SCFAs, assessed GI symptoms and analyzed the relationship between the gut microbiome and fecal SCFAs in autistic and neurotypical individuals. The results showed that the compositions of the gut microbiota and SCFAs were altered in ASD individuals. We found lower levels of fecal acetic acid and butyrate and a higher level of fecal valeric acid in ASD subjects. We identified decreased abundances of key butyrate-producing taxa (Ruminococcaceae, Eubacterium, Lachnospiraceae and Erysipelotrichaceae) and an increased abundance of valeric acid associated bacteria (Acidobacteria) among autistic individuals. Constipation was the only GI disorder in ASD children in the present study. We also found enriched Fusobacterium, Barnesiella, Coprobacter and valeric acid-associated bacteria (Actinomycetaceae) and reduced butyrate-producing taxa in constipated autistic subjects. It is suggested that the gut microbiota contributes to fecal SCFAs and constipation in autism. Modulating the gut microbiota, especially butyrate-producing bacteria, could be a promising strategy in the search for alternatives for the treatment of autism spectrum disorder.

The relative contributions of apoptosis and necrosis in brain injury have been a matter of much debate. Caspase-3 has been identified as a key protease in the execution of apoptosis, whereas calpains have mainly been implicated in excitotoxic neuronal injury. In a model of unilateral hypoxia-ischemia in 7-day-old rats, caspase-3-like activity increased 16-fold 24 h postinsult, coinciding with cleavage of the caspase-3 proenzyme and endogenous caspase-3 substrates. This activation was significantly decreased by pharmacological calpain inhibition, using CX295, a calpain inhibitor that did not inhibit purified caspase-3 in vitro. Activation of caspase-3 by m-calpain, but not μ-calpain, was facilitated in a dose-dependent manner in vitroby incubating cytosolic fractions, containing caspase-3 proform, with calpains. This facilitation required the presence of some active caspase-3 and could be abolished by including the specific calpain inhibitor calpastatin. This indicates that initial cleavage of caspase-3 by m-calpain, producing a 29-kDa fragment, facilitates the subsequent cleavage into active forms. This is the first report to our knowledge suggesting a direct link between the early, excitotoxic, calcium-mediated activation of calpain after cerebral hypoxia-ischemia and the subsequent activation of caspase-3, thus representing a tentative pathway of “pathological apoptosis.” The relative contributions of apoptosis and necrosis in brain injury have been a matter of much debate. Caspase-3 has been identified as a key protease in the execution of apoptosis, whereas calpains have mainly been implicated in excitotoxic neuronal injury. In a model of unilateral hypoxia-ischemia in 7-day-old rats, caspase-3-like activity increased 16-fold 24 h postinsult, coinciding with cleavage of the caspase-3 proenzyme and endogenous caspase-3 substrates. This activation was significantly decreased by pharmacological calpain inhibition, using CX295, a calpain inhibitor that did not inhibit purified caspase-3 in vitro. Activation of caspase-3 by m-calpain, but not μ-calpain, was facilitated in a dose-dependent manner in vitroby incubating cytosolic fractions, containing caspase-3 proform, with calpains. This facilitation required the presence of some active caspase-3 and could be abolished by including the specific calpain inhibitor calpastatin. This indicates that initial cleavage of caspase-3 by m-calpain, producing a 29-kDa fragment, facilitates the subsequent cleavage into active forms. This is the first report to our knowledge suggesting a direct link between the early, excitotoxic, calcium-mediated activation of calpain after cerebral hypoxia-ischemia and the subsequent activation of caspase-3, thus representing a tentative pathway of “pathological apoptosis.” hypoxia-ischemia aminomethylcoumarin DNA fragmentation factor 45 α-fodrin breakdown product inhibitor of caspase-activated DNase poly(ADP-ribose) polymerase dithiothreitol 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone t-butoxycarbonyl-Asp-(OMe)-fluoromethyl ketone phosphate-buffered saline polymerase chain reaction glyceraldehyde-3-phosphate dehydrogenase Leu-Tyr-AMC The relative contributions of necrosis and apoptosis to the injury that develops after cerebral hypoxia-ischemia (HI)1 has been a matter of much debate (1Lee J.M. Zipfel G.J. Choi D.W. Nature. 1999; 399: 7-14Crossref PubMed Scopus (1004) Google Scholar). Recent studies suggest that cell death after HI is different from developmentally regulated cell death in most cases and cannot appropriately be described as apoptotic (2MacManus J.P. Fliss H. Preston E. Rasquinha I. Tuor U. J. Cereb. Blood Flow Metab. 1999; 19: 502-510Crossref PubMed Scopus (38) Google Scholar, 3Jin K. Chen J. Nagayama T. Chen M. Sinclair J. Graham S.H. Simon R.P. J. Neurochem. 1999; 72: 1204-1214Crossref PubMed Scopus (69) Google Scholar, 4Ishimaru M.J. Ikonomidou C. Tenkova T.I. Der T.C. Dikranian K. Sesma M.A. Olney J.W. J. Comp. Neurol. 1999; 408: 461-476Crossref PubMed Scopus (188) Google Scholar, 5Portera-Cailliau C. Price D.L. Martin L.J. J. Comp. Neurol. 1997; 378: 70-87PubMed Google Scholar). Nevertheless, HI cell death shares important morphological and biochemical features with apoptotic cell death, such as activation of caspases and nucleosomal DNA fragmentation (6Zhu C. Wang X. Hagberg H. Blomgren K. J. Neurochem. 2000; 75: 819-829Crossref PubMed Scopus (111) Google Scholar, 7Linnik M.D. Zobrist R.H. Hatfield M.D. Stroke. 1993; 24: 2002-2008Crossref PubMed Scopus (675) Google Scholar, 8MacManus J.P. Buchan A.M. Hill I.E. Rasquinha I. Preston E. Neurosci. Lett. 1993; 164: 89-92Crossref PubMed Scopus (450) Google Scholar, 9Cheng Y. Deshmukh M. DaCosta A. Demaro J.A. Gidday J.M. Shah A. Sun Y. Jacquin M.F. Johnson E.M. Holtzman D.M. J. Clin. Invest. 1998; 101: 1992-1999Crossref PubMed Scopus (481) Google Scholar, 10Namura S. Zhu J. Fink K. Endres M. Srinivasan A. Tomaselli K.J. Yuan J. Moskowitz M.A. J. Neurosci. 1998; 18: 3659-3668Crossref PubMed Google Scholar, 11Chen J. Nagayama T. Jin K. Stetler R.A. Zhu R.L. Graham S.H. Simon R.P. J. Neurosci. 1998; 18: 4914-4928Crossref PubMed Google Scholar, 12Pulera M.R. Adams L.M. Liu H. Santos D.G. Nishimura R.N. Yang F. Cole G.M. Wasterlain C.G. Stroke. 1998; 29: 2622-2630Crossref PubMed Scopus (180) Google Scholar, 13Ni B. Wu X. Su Y. Stephenson D. Smalstig E.B. Clemens J. Paul S.M. J. Cereb. Blood Flow Metab. 1998; 18: 248-256Crossref PubMed Scopus (147) Google Scholar, 14Ma J. Endres M. Moskowitz M.A. Br. J. Pharmacol. 1998; 124: 756-762Crossref PubMed Scopus (156) Google Scholar, 15Endres M. Namura S. Shimizu-Sasamata M. Waeber C. Zhang L. Gomez-Isla T. Hyman B.T. Moskowitz M.A. J. Cereb. Blood Flow Metab. 1998; 18: 238-247Crossref PubMed Scopus (509) Google Scholar, 16Hara H. Friedlander R.M. Gagliardini V. Ayata C. Fink K. Huang Z. Shimizu-Sasamata M. Yuan J. Moskowitz M.A. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2007-2012Crossref PubMed Scopus (798) Google Scholar, 17Yue X. Mehmet H. Penrice J. Cooper C. Cady E. Wyatt J.S. Reynolds E.O. Edwards A.D. Squier M.V. Neuropathol. Appl. Neurobiol. 1997; 23: 16-25Crossref PubMed Scopus (177) Google Scholar). Caspases, a family of cysteine proteases with an unusual substrate specificity, requiring an aspartate residue in the P1 position, have been identified as key executors of apoptosis (18Stennicke H.R. Salvesen G.S. Biochim. Biophys. Acta. 2000; 1477: 299-306Crossref PubMed Scopus (295) Google Scholar). Calpains, another family of cysteine proteases, are calcium-activated and are proposed to participate in the turnover of cytoskeletal proteins and regulation of kinases, transcription factors, and receptors (19Chan S.L. Mattson M.P. J. Neurosci. Res. 1999; 58: 167-190Crossref PubMed Scopus (372) Google Scholar, 20Sorimachi H. Ishiura S. Suzuki K. Biochem. J. 1997; 328: 721-732Crossref PubMed Scopus (620) Google Scholar). Calpains have mainly been implicated in excitotoxic neuronal injury and necrosis (21Seubert P. Lee K. Lynch G. Brain Res. 1989; 492: 366-370Crossref PubMed Scopus (189) Google Scholar, 22Siman R. Noszek J. Neuron. 1988; 1: 279-287Abstract Full Text PDF PubMed Scopus (381) Google Scholar, 23Bednarski E. Vanderklish P. Gall C. Saido T. Bahr B. Lynch G. Brain Res. 1995; 694: 147-157Crossref PubMed Scopus (61) Google Scholar). Pharmacological inhibitors of calpains and caspases exert cerebroprotective effects (9Cheng Y. Deshmukh M. DaCosta A. Demaro J.A. Gidday J.M. Shah A. Sun Y. Jacquin M.F. Johnson E.M. Holtzman D.M. J. Clin. Invest. 1998; 101: 1992-1999Crossref PubMed Scopus (481) Google Scholar, 14Ma J. Endres M. Moskowitz M.A. Br. J. Pharmacol. 1998; 124: 756-762Crossref PubMed Scopus (156) Google Scholar, 15Endres M. Namura S. Shimizu-Sasamata M. Waeber C. Zhang L. Gomez-Isla T. Hyman B.T. Moskowitz M.A. J. Cereb. Blood Flow Metab. 1998; 18: 238-247Crossref PubMed Scopus (509) Google Scholar, 16Hara H. Friedlander R.M. Gagliardini V. Ayata C. Fink K. Huang Z. Shimizu-Sasamata M. Yuan J. Moskowitz M.A. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2007-2012Crossref PubMed Scopus (798) Google Scholar, 24Lee K. Frank S. Vanderklish P. Arai A. Lynch G. Proc. Natl. Acad. Sci. U. S. A. 1991; 88: 7233-7237Crossref PubMed Scopus (333) Google Scholar, 25Rami A. Krieglstein J. Brain Res. 1993; 609: 67-70Crossref PubMed Scopus (164) Google Scholar, 26Bartus R. Hayward N. Elliott P. Sawyer S. Baker K. Dean R. Akiyama A. Straub J. Harbeson S. Li Z. Powers J. Stroke. 1994; 25: 2265-2270Crossref PubMed Scopus (198) Google Scholar). A growing body of literature has emerged, demonstrating functional connections between calpains and caspases (27Wang K.K. Trends Neurosci. 2000; 23: 20-26Abstract Full Text Full Text PDF PubMed Scopus (811) Google Scholar). Common substrate proteins have been identified, such as fodrin (28Nath R. Raser K. Stafford D. Hajimohammadreza I. Posner A. Allen H. Talanian R. Yuen P. Gilbertsen R. Wang K. Biochem. J. 1996; 319: 683-690Crossref PubMed Scopus (398) Google Scholar, 29Jänicke R. Ng P. Sprengart M. Porter A. J. Biol. Chem. 1998; 273: 15540-15545Abstract Full Text Full Text PDF PubMed Scopus (444) Google Scholar, 30Vanags D. Pörn-Ares M. Coppola S. Burgess D. Orrenius S. J. Biol. Chem. 1996; 271: 31075-31085Abstract Full Text Full Text PDF PubMed Scopus (356) Google Scholar, 31Wang K. Posmantur R. Nath R. McGinnis K. Whitton M. Talanian R. Galntz S. Morrow J. J. Biol. Chem. 1998; 273: 22490-22497Abstract Full Text Full Text PDF PubMed Scopus (283) Google Scholar), calpastatin (32Pörn-Ares M.I. Samali A. Orrenius S. Cell Death Differ. 1998; 5: 1028-1033Crossref PubMed Scopus (184) Google Scholar, 33Wang K.K. Posmantur R. Nadimpalli R. Nath R. Mohan P. Nixon R.A. Talanian R.V. Keegan M. Herzog L. Allen H. Arch. Biochem. Biophys. 1998; 356: 187-196Crossref PubMed Scopus (231) Google Scholar), actin (34Villa P.G. Henzel W.J. Sensenbrenner M. Henderson C.E. Pettmann B. J. Cell Sci. 1998; 111: 713-722PubMed Google Scholar), PARP (35McGinnis N. Wang K.K. Biochem. Biophys. Res. 1999; PubMed Scopus Google Scholar), and N. L. C. C. A.M. M. A. A. P. J. Neurosci. 1998; 18: PubMed Google Scholar). are demonstrating cleavage of caspase-3 (35McGinnis N. Wang K.K. Biochem. Biophys. Res. 1999; PubMed Scopus Google Scholar, H.R. H. Salvesen G.S. 1999; 94: PubMed Google and A. G. J. 1999; 18: PubMed Scopus Google Scholar, B.T. K. Li P. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google as as and B.T. K. Li P. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). the was by calpain apoptosis of A. Y. 1998; PubMed Scopus Google Scholar), and calpain be the in an into a T. Yuan J. J. Cell Biol. 2000; PubMed Scopus Google Scholar). between calpains and the of caspases in apoptosis of B. J. J. Biol. Chem. 1998; 273: Full Text Full Text PDF PubMed Scopus Google Scholar), whereas studies an calpains in the apoptosis of A.M. E. J. 1999; PubMed Scopus Google and M. J. J. 1997; Google Scholar). and Yuan that be the activation of by the a link between and apoptosis T. Yuan J. J. Cell Biol. 2000; PubMed Scopus Google Scholar). that the of calpastatin in a model of cerebral HI a K. U. M. Bahr A. Saido T.C. S. Hagberg H. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar), the the activation of demonstrating of calpastatin by caspase-3 (32Pörn-Ares M.I. Samali A. Orrenius S. Cell Death Differ. 1998; 5: 1028-1033Crossref PubMed Scopus (184) Google Scholar, 33Wang K.K. Posmantur R. Nadimpalli R. Nath R. Mohan P. Nixon R.A. Talanian R.V. Keegan M. Herzog L. Allen H. Arch. Biochem. Biophys. 1998; 356: 187-196Crossref PubMed Scopus (231) Google to the and activation of proteases and in HI was in 7-day-old of J. R. J. Neurol. PubMed Scopus Google Scholar, H. E. N. P. Biol. 1994; PubMed Scopus Google Scholar). The with and in a of and and the of was The was between of the the with a and the to The in a with a in The in the was the to and but not to was by the of with the calpain inhibitor The first of in to body was after with of the to body h 24 with The by and the a of in and and to the was from The was by in of containing and with an of and active caspase-3 of of caspase-3 is not was with of protease inhibitor and with of substrate in containing and of was using a with an of and an of cleavage was from and as relative S. C. E. Biochem. J. PubMed Scopus Google of from the in of was with of inhibitor and with of in containing and cleavage was as described the caspase-3 as from and benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone ketone from of with of containing and a of in inhibitors but with was of was as described and as of of and The caspase-3 and the of caspase-3 the a from The the active of caspase-3 in (6Zhu C. Wang X. Hagberg H. Blomgren K. J. Neurochem. 2000; 75: 819-829Crossref PubMed Scopus (111) Google Scholar). A that the breakdown product of α-fodrin was to fodrin cleavage B. S. G. Lynch G. J. Pharmacol. 1995; 273: Google Scholar). The was and was by with a X. H. C. Wang X. 1997; Full Text Full Text PDF PubMed Scopus Google from of The The PARP fodrin and and from The of the fodrin was by incubating a with caspase-3 The and the to and In the with m-calpain, a with In the with caspase-3, could be using the of fodrin was as using the that the fodrin as as cleavage The of in was to J. K. C. M. Biochem. 1994; PubMed Scopus Google Scholar). to of and to using and and using the in a and using the was and the was as of was by in with the was to between S. Zhu J. Fink K. Endres M. Srinivasan A. Tomaselli K.J. Yuan J. Moskowitz M.A. J. Neurosci. 1998; 18: 3659-3668Crossref PubMed Google Scholar). and with in The and 24 with and the and into in and in in a of in and as with in was by the in was with in was in and by in was using and as The was in containing and by in was using 24 and of after and The and in was from P. N. Biochem. PubMed Scopus Google Scholar), and was with the and and to 24 The was and of first of and of of and and by was and the reaction was to by of The thus was to with and subsequent of a of and a of of DNA polymerase and of as caspase-3 and and from caspase-3 was as 24 of was as of The and such that the caspase-3 and the be in the of and of not The caspase-3 and with and The and the using the The relative of caspase-3 was after to to the of the of the of in of and 45 to cytosolic of of of of of the in in the and of m-calpain, caspase-3, purified calpastatin calpastatin was in some the inhibitors with the to the The by of of and to of to and The to and Biochem. PubMed Scopus Google Scholar), using a calpastatin was purified from as described S. C. E. Biochem. J. PubMed Scopus Google and purified acid and B. J. Blomgren K. J. Arch. 1988; PubMed Scopus Google Scholar). of the was of active caspase-3 is as the of caspase-3 that of in the described to caspase-3 could the activity activity was as described of with activity was in brain and increased in the with the in with (9Cheng Y. Deshmukh M. DaCosta A. Demaro J.A. Gidday J.M. Shah A. Sun Y. Jacquin M.F. Johnson E.M. Holtzman D.M. J. Clin. Invest. 1998; 101: 1992-1999Crossref PubMed Scopus (481) Google Scholar), and a 24 h The between the and was significantly decreased after calpain inhibition, from in the to in the of in calpain the activation of caspase-3-like activity cleavage of the caspase-3 and of endogenous caspase-3 substrate after with the calpain inhibitor h 24 h The between the and the is and the cleavage product of caspase-3 the is that between the and the in the could not be in the in a The caspase-3-like activity cleavage of the caspase-3 and of endogenous caspase-3 substrate after with the calpain inhibitor h 24 h The between the and the is and the cleavage product of caspase-3 the is that between the and the in the could not be in the The caspase-3 a with of and The of caspase-3 as the brain the the The was after HI h to h after the with of and in the The of the and to a of the as by a decreased This of the to a 24 h a the activity The 29-kDa was in from to h whereas the was in mainly 24 h was with some the and a The activation of caspase-3, as by the and of the proform, was in by the of the and the by the of the and in the The of the between the and the was significantly increased after calpain inhibition, from in the to in the The between the and in the a from in the to in the The of the active cleavage of the the caspase-3-like activity was with the and the 29-kDa from the 24 h The between the and the a with the activity and did the 29-kDa The a caspase-3 and a fragment, as The most after to was that the of caspase-3 an from to to the The by The of was to and m-calpain, but did not inhibit caspase-3 The of a inhibitor of was calpains caspase-3 another caspase-3 much calpains the protease inhibitors inhibitor the proteases activity in are of the inhibitors the active cysteine but with different was to caspase-3, and was to and in a The inhibitor the proteases activity in are of the inhibitors the active cysteine but with different was to caspase-3, and was to and The of and PARP a 24 h with the activity the and of but specific could be using The of in the to a decreased and between the and the This was significantly increased after calpain inhibition, from in the to in the of the PARP was by an increased of fodrin the and as as the cleavage The caspase-3 was to be specific caspase-3 in and was in with as by the of and with DNA (6Zhu C. Wang X. Hagberg H. Blomgren K. J. Neurochem. 2000; 75: 819-829Crossref PubMed Scopus (111) Google Scholar). The of by the increased and was 24 h h A and The in of but a of after HI A and of caspase-3 and of and active is of using the active caspase-3 and and 24 h of demonstrating the of caspase-3 and calpain activity 24 h of the calpain activity has but the of active caspase-3 has of did not the cleavage the caspase-3 A and was a 29-kDa to the after HI in This did not an of was was with calpastatin purified CX295, HI in was the the of the that was and This was not by calpain purified by but could be by and calpain The caspase-3-like activity increased after of from different with and could be abolished calpastatin was in the was in the the activity was not significantly increased of active caspase-3 to cytosolic fractions, the active caspase-3 could be as a as an of active caspase-3 the of the 29-kDa from the The of the 29-kDa was by including of active caspase-3 to the the activity increased caspase-3 was in the of This could be abolished by including in the Caspase-3 could not m-calpain, incubating with of caspase-3 did not the calpain activity between and A cytosolic containing caspase-3 proform, was with and different of caspase-3 and as active caspase-3 was the of a in the demonstrating the the relative in caspase-3-like activity in the is in the a caspase-3 that the active caspase-3 be as a as the of active caspase-3 an increased of the 29-kDa but was active as that the some endogenous m-calpain, in to caspase-3 a demonstrating that the of caspase-3 cleavage in the was not to is that in a producing an the The cleavage of the from to was could be by and calpastatin not inhibit m-calpain, but not μ-calpain, could cleavage of the of producing a 29-kDa could be This is the report to our knowledge demonstrating a functional in substrate between the calpains. Recent studies have cleavage of caspase-3 by producing an cleavage product (35McGinnis N. Wang K.K. Biochem. Biophys. Res. 1999; PubMed Scopus Google Scholar, H.R. H. Salvesen G.S. 1999; 94: PubMed Google Scholar), but another report to such cleavage A. G. J. 1999; 18: PubMed Scopus Google Scholar). H.R. H. Salvesen G.S. 1999; 94: PubMed Google identified the cleavage in the of In was not m-calpain, of could functional that calpains are to and that calpains as of B.T. K. Li P. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). of studies calpastatin to the the first to that calpains H.R. H. Salvesen G.S. 1999; 94: PubMed Google Scholar), is important to in using as a that was calpains and that was with to between calpain and the effects cysteine proteases have not been purified caspase-3 was not to caspase-3 is the cytosolic from containing of This that and but the cannot be that have been the The of a and and is that was the that was HI in This indicates that are of the caspase-3 and that the of to active is different in is not the are the of The caspase-3-like activity was with the of the and the that the 29-kDa be an in the of the active The that of 29-kDa caspase-3 activity indicates that the to the active be facilitated by initial cleavage in the This was by the that and caspase-3 to be active to that the initial of the 29-kDa was by cleavage into the active was that the was to activation of m-calpain, the activity of the activity was could not be by the of active caspase-3 The was by that a of caspase-3, with a of m-calpain, the of the 29-kDa as as the caspase-3 was The activation of first calpain and caspase-3 after HI was to in the and in of of This with an model of of caspase-3 activation the of neuronal has been that calpain activation after in an the and a coinciding with (21Seubert P. Lee K. Lynch G. Brain Res. 1989; 492: 366-370Crossref PubMed Scopus (189) Google Scholar, T. M. K. E. Y. S. J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar). This of fodrin was in our model K. S. Saido T. J. A. Hagberg H. Brain Res. 1995; PubMed Scopus Google Scholar), but the of calpastatin was K. U. M. Bahr A. Saido T.C. S. Hagberg H. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar). is not calpains caspase-3, the calpastatin have been to (32Pörn-Ares M.I. Samali A. Orrenius S. Cell Death Differ. 1998; 5: 1028-1033Crossref PubMed Scopus (184) Google Scholar, 33Wang K.K. Posmantur R. Nadimpalli R. Nath R. Mohan P. Nixon R.A. Talanian R.V. Keegan M. Herzog L. Allen H. Arch. Biochem. Biophys. 1998; 356: 187-196Crossref PubMed Scopus (231) Google Scholar). The caspase-3 activity in was and is that activation of caspase-3 after HI is different from that developmentally regulated are of increased caspase-3 a after cerebral 7Linnik M.D. Zobrist R.H. Hatfield M.D. Stroke. 1993; 24: 2002-2008Crossref PubMed Scopus (675) Google Scholar, 8MacManus J.P. Buchan A.M. Hill I.E. Rasquinha I. Preston E. Neurosci. Lett. 1993; 164: 89-92Crossref PubMed Scopus (450) Google Scholar, and 10Namura S. Zhu J. Fink K. Endres M. Srinivasan A. Tomaselli K.J. Yuan J. Moskowitz M.A. J. Neurosci. 1998; 18: 3659-3668Crossref PubMed Google Scholar, 11Chen J. Nagayama T. Jin K. Stetler R.A. Zhu R.L. Graham S.H. Simon R.P. J. Neurosci. 1998; 18: 4914-4928Crossref PubMed Google Scholar, 12Pulera M.R. Adams L.M. Liu H. Santos D.G. Nishimura R.N. Yang F. Cole G.M. Wasterlain C.G. Stroke. 1998; 29: 2622-2630Crossref PubMed Scopus (180) Google as as of after of inhibitors 9Cheng Y. Deshmukh M. DaCosta A. Demaro J.A. Gidday J.M. Shah A. Sun Y. Jacquin M.F. Johnson E.M. Holtzman D.M. J. Clin. Invest. 1998; 101: 1992-1999Crossref PubMed Scopus (481) Google and 14Ma J. Endres M. Moskowitz M.A. Br. J. Pharmacol. 1998; 124: 756-762Crossref PubMed Scopus (156) Google Scholar, 15Endres M. Namura S. Shimizu-Sasamata M. Waeber C. Zhang L. Gomez-Isla T. Hyman B.T. Moskowitz M.A. J. Cereb. Blood Flow Metab. 1998; 18: 238-247Crossref PubMed Scopus (509) Google Scholar, 16Hara H. Friedlander R.M. Gagliardini V. Ayata C. Fink K. Huang Z. Shimizu-Sasamata M. Yuan J. Moskowitz M.A. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2007-2012Crossref PubMed Scopus (798) Google Scholar), but the that to after the a link between and “pathological apoptosis.” This is by our that the caspase-3 activation after HI M. U. Zhu C. Wang X. Blomgren K. Hagberg H. 2000; PubMed Scopus Google Scholar). studies have that to the activation of T. Zhu H. N. Li E. J. Yuan J. Nature. 2000; PubMed Scopus Google Scholar), by T. Yuan J. J. Cell Biol. 2000; PubMed Scopus Google Scholar). Liu Y. Blomgren K. J. Cereb. Blood Flow Metab. 2000; PubMed Scopus Google that of active caspase-3 in after but in B. Wu X. Su Y. Stephenson D. Smalstig E.B. Clemens J. Paul S.M. J. Cereb. Blood Flow Metab. 1998; 18: 248-256Crossref PubMed Scopus (147) Google of caspase-3 in the and brain but in the Chen J. Nagayama T. Jin K. Stetler R.A. Zhu R.L. Graham S.H. Simon R.P. J. Neurosci. 1998; 18: 4914-4928Crossref PubMed Google of caspase-3 and in the of an in the after h of after in the with of caspase-3 and but the the to be after HI in the The of calpains K. J. Neurochem. Res. 1989; PubMed Scopus Google and caspase-3 is in the that proteases be important in the In is the first report to our knowledge demonstrating a functional in the substrate of the calpains and as as facilitated activation of caspase-3 by Caspase-3 has been identified as a key protease in the execution of apoptosis, whereas calpains have mainly been implicated in excitotoxic neuronal injury. suggest a direct link between the early, excitotoxic, calcium-mediated activation of calpains after cerebral HI and the subsequent activation of caspase-3, thus representing a tentative pathway of “pathological apoptosis.” This be important in the brain of the of calpains and caspase-3 are to the active caspase-3 and to Wang of the

OBJECTIVE: The purpose of this study was to evaluate the efficacy and safety of erythropoietin in neonatal hypoxic-ischemic encephalopathy (HIE), by using a randomized, prospective study design. METHODS: A total of 167 term infants with moderate/severe HIE were assigned randomly to receive either erythropoietin (N = 83) or conventional treatment (N = 84). Recombinant human erythropoietin, at either 300 U/kg (N = 52) or 500 U/kg (N = 31), was administered every other day for 2 weeks, starting <48 hours after birth. The primary outcome was death or disability. Neurodevelopmental outcomes were assessed at 18 months of age. RESULTS: Complete outcome data were available for 153 infants. Nine patients dropped out during treatment, and 5 patients were lost to follow-up monitoring. Death or moderate/severe disability occurred for 35 (43.8%) of 80 infants in the control group and 18 (24.6%) of 73 infants in the erythropoietin group (P = .017) at 18 months. The primary outcomes were not different between the 2 erythropoietin doses. Subgroup analyses indicated that erythropoietin improved long-term outcomes only for infants with moderate HIE (P = .001) and not those with severe HIE (P = .227). No negative hematopoietic side effects were observed. CONCLUSION: Repeated, low-dose, recombinant human erythropoietin treatment reduced the risk of disability for infants with moderate HIE, without apparent side effects.

Apoptosis-inducing factor (AIF) triggers apoptosis in a caspase-independent manner. Here we report for the first time involvement of AIF in neuronal death induced by cerebral ischemia. Unilateral cerebral hypoxia-ischemia (HI) was induced in 7-day-old rats by ligation of the left carotid artery and hypoxia (7.7% O2) for 55 min. AIF release from mitochondria and AIF translocation to nuclei was detected immediately after HI, and only in damaged areas, as judged by the concurrent loss of MAP-2. AIF release was detected earlier than that of cytochrome c. Cells with AIF-positive nuclei displayed nuclear condensation and signs of DNA damage. The number of AIF-positive nuclei showed a positive correlation with the infarct volume 72 h post-HI, and this was not changed by treating the animals with boc-Asp-fmk (BAF), a multicaspase inhibitor. BAF treatment reduced the activity of caspase-3, -2 and -9 (78, 73 and 33%, respectively), and prevented caspase-dependent fodrin cleavage in vivo, but did not affect AIF release from mitochondria or the frequency of positive nuclear AIF or DNA damage 72 h post-HI, indicating that these processes occurred in a caspase-independent fashion. In summary, AIF-mediated cell death may be an important mechanism of HI-induced neuronal loss in the immature brain.

Isoflurane and related anesthetics are widely used to anesthetize children, ranging from premature babies to adolescents. Concerns have been raised about the safety of these anesthetics in pediatric patients, particularly regarding possible negative effects on cognition. The purpose of this study was to investigate the effects of repeated isoflurane exposure of juvenile and mature animals on cognition and neurogenesis. Postnatal day 14 (P14) rats and mice, as well as adult (P60) rats, were anesthetized with isoflurane for 35 mins daily for four successive days. Object recognition, place learning and reversal learning as well as cell death and cytogenesis were evaluated. Object recognition and reversal learning were significantly impaired in isoflurane-treated young rats and mice, whereas adult animals were unaffected, and these deficits became more pronounced as the animals grew older. The memory deficit was paralleled by a decrease in the hippocampal stem cell pool and persistently reduced neurogenesis, subsequently causing a reduction in the number of dentate gyrus granule cell neurons in isoflurane-treated rats. There were no signs of increased cell death of progenitors or neurons in the hippocampus. These findings show a previously unknown mechanism of neurotoxicity, causing cognitive deficits in a clearly age-dependent manner.

Sex-related brain injury was evaluated after unilateral hypoxia-ischaemia (HI) in C57/BL6 mice on postnatal day (P) 5, 9, 21 or 60, corresponding developmentally to premature, term, juvenile and adult human brains. There was no sex difference in brain injury when the insult was severe, as evaluated by pathological scoring or tissue loss, but when the insult was moderate, adult (P60) females displayed less injury. In the immature (P9) male brains, neurones displayed a more pronounced translocation of apoptosis-inducing factor (AIF) (loss of AIF from the mitochondrial fraction and increase in nuclear AIF) after HI, whereas the female brain neurones displayed a stronger activation of caspase 3 (more pronounced loss of pro-caspase 3, increase in cleaved caspase 3 and increase in caspase 3 enzymatic activity). Two other mechanisms of injury, peroxynitrite-induced formation of nitrotyrosine and autophagy, were no different between males and females at P9. These data show that the CNS is more resistant to HI in adult females compared with males, whereas no sex differences were found in the extent of injury in neonatal mice. However, critical sex-dependent differences were demonstrated in vivo with regard to cellular, apoptosis-related mechanisms.

Inhibition of matrix metalloproteinase-9 (MMP-9) protects the adult brain after cerebral ischemia. However, the role of MMP-9 in the immature brain after hypoxia-ischemia (HI) is unknown. We exposed MMP-9(-/-) [MMP-9 knock-out (KO)] and wild-type (WT) mice to HI on postnatal day 9. HI was induced by unilateral ligation of the left carotid artery followed by hypoxia (10% O2; 36 degrees C). Gelatin zymography showed that MMP-9 activity was transiently increased at 24 h after HI in the ipsilateral hemisphere and MMP-9-positive cells were colocalized with activated microglia. Seven days after 50 min of HI, cerebral tissue volume loss was reduced in MMP-9 KO (21.8 +/- 1.7 mm3; n = 22) compared with WT (32.3 +/- 2.1 mm3; n = 22; p < 0.001) pups, and loss of white-matter components was reduced in MMP-9 KO compared with WT pups (neurofilament: WT, 50.9 +/- 5.4%; KO, 18.4 +/- 3.1%; p < 0.0001; myelin basic protein: WT, 57.5 +/- 5.8%; KO, 23.2 +/- 3.5%; p = 0.0001). The neuropathological changes were associated with a delayed and diminished leakage of the blood-brain barrier (BBB) and a decrease in inflammation in MMP-9-deficient animals. In contrast, the neuroprotective effects after HI in MMP-9-deficient animals were not linked to either caspase-dependent (caspase-3 and cytochrome c) or caspase-independent (apoptosis-inducing factor) processes. This study demonstrates that excessive activation of MMP-9 is deleterious to the immature brain, which is associated with the degree of BBB leakage and inflammation. In contrast, apoptosis does not appear to be a major contributing factor.

Inactive von Hippel-Lindau (VHL) is linked to metabolic reprogramming and plays pivotal roles in the pathogenesis of clear cell renal cell carcinoma (ccRCC). Here, we identify a previously unknown oncogenic role for inactive VHL in actively triggering histone lactylation to promote ccRCC progression. In patients with ccRCC, inactive VHL positively correlates with the presence of histone lactylation, and high levels of histone lactylation indicates poor patient prognosis. Inactive VHL-triggered histone lactylation contributes to ccRCC progression by activating the transcription of platelet-derived growth factor receptor β (PDGFRβ). In turn, PDGFRβ signaling is shown to stimulate histone lactylation, thereby forming an oncogenic positive feedback loop in ccRCC. Target correction of aberrant histone lactylation represses the growth and metastasis of ccRCC in vivo. More importantly, the combined inhibition of histone lactylation and PDGFRβ significantly reinforces the therapeutic efficacy. This work underscores the importance of histone lactylation in facilitating ccRCC progression and suggests targeting the positive feedback loop between histone lactylation and PDGFRβ signaling might provide a promising therapeutic strategy for ccRCC patients.

Upon cerebral hypoxia-ischemia (HI), apoptosis-inducing factor (AIF) can move from mitochondria to nuclei, participate in chromatinolysis, and contribute to the execution of cell death. Previous work (Cande, C., N. Vahsen, I. Kouranti, E. Schmitt, E. Daugas, C. Spahr, J. Luban, R.T. Kroemer, F. Giordanetto, C. Garrido, et al. 2004. Oncogene. 23:1514-1521) performed in vitro suggests that AIF must interact with cyclophilin A (CypA) to form a proapoptotic DNA degradation complex. We addressed the question as to whether elimination of CypA may afford neuroprotection in vivo. 9-d-old wild-type (WT), CypA(+/-), or CypA(-/-) mice were subjected to unilateral cerebral HI. The infarct volume after HI was reduced by 47% (P = 0.0089) in CypA(-/-) mice compared with their WT littermates. Importantly, CypA(-/-) neurons failed to manifest the HI-induced nuclear translocation of AIF that was observed in WT neurons. Conversely, CypA accumulated within the nuclei of damaged neurons after HI, and this nuclear translocation of CypA was suppressed in AIF-deficient harlequin mice. Immunoprecipitation of AIF revealed coprecipitation of CypA, but only in injured, ischemic tissue. Surface plasmon resonance revealed direct molecular interactions between recombinant AIF and CypA. These data indicate that the lethal translocation of AIF to the nucleus requires interaction with CypA, suggesting a model in which two proteins that normally reside in separate cytoplasmic compartments acquire novel properties when moving together to the nucleus.

BACKGROUND: Aberrant expression of circular RNA contributes to human diseases. Circular RNAs regulate gene expression by sequestering specific microRNAs. In this study, we investigated whether circMAP3K5 (circular mitogen-activated protein kinase 5) could act as a competing endogenous microRNA-22-3p (miR-22-3p) sponge and regulate neointimal hyperplasia. METHODS: Circular RNA profiling from genome-wide RNA sequencing data was compared between human coronary artery smooth muscle cells (SMCs) treated with or without platelet-derived growth factor. Expression levels of circMAP3K5 were assessed in human coronary arteries from autopsies on patients with dilated cardiomyopathy or coronary heart disease. The role of circMAP3K5 in intimal hyperplasia was further investigated in mice with adeno-associated virus 9-mediated circMAP3K5 transfection. SMC-specific Tet2 (ten-eleven translocation-2) knockout mice and global miR-22-3p knockout mice were used to delineate the mechanism by which circMAP3K5 attenuated neointimal hyperplasia using the femoral arterial wire injury model. RESULTS: RNA sequencing demonstrated that treatment with platelet-derived growth factor-BB significantly reduced expression of circMAP3K5 in human coronary artery SMCs. Wire-injured mouse femoral arteries and diseased arteries from patients with coronary heart disease (where platelet-derived growth factor-BB is increased) confirmed in vivo downregulation of circMAP3K5 associated with injury and disease. Lentivirus-mediated overexpression of circMAP3K5 inhibited the proliferation of human coronary artery SMCs. In vivo adeno-associated virus 9-mediated transfection of circMap3k5 (mouse circular Map3k5) specifically inhibited SMC proliferation in the wire-injured mouse arteries, resulting in reduced neointima formation. Using a luciferase reporter assay and RNA pull-down, circMAP3K5 (human circular MAP3K5) was found to sequester miR-22-3p, which, in turn, inhibited the expression of TET2. Both in vitro and in vivo results demonstrate that the loss of miR-22-3p recapitulated the antiproliferative effect of circMap3k5 on vascular SMCs. In SMC-specific Tet2 knockout mice, loss of Tet2 abolished the circMap3k5-mediated antiproliferative effect on vascular SMCs. CONCLUSIONS: We identify circMAP3K5 as a master regulator of TET2-mediated vascular SMC differentiation. Targeting the circMAP3K5/miR-22-3p/TET2 axis may provide a potential therapeutic strategy for diseases associated with intimal hyperplasia, including restenosis and atherosclerosis.

Antibiotic resistance is currently one of the greatest threats to human health. Widespread use and residues of antibiotics in humans, animals, and the environment can exert selective pressure on antibiotic resistance bacteria (ARB) and antibiotic resistance gene (ARG), accelerating the flow of antibiotic resistance. As ARG spreads to the population, the burden of antibiotic resistance in humans increases, which may have potential health effects on people. Therefore, it is critical to mitigate the spread of antibiotic resistance to humans and reduce the load of antibiotic resistance in humans. This review briefly described the information of global antibiotic consumption information and national action plans (NAPs) to combat antibiotic resistance and provided a set of feasible control strategies for the transmission of ARB and ARG to humans in three areas including (a) Reducing the colonization capacity of exogenous ARB, (b) Enhancing human colonization resistance and mitigating the horizontal gene transfer (HGT) of ARG, (c) Reversing ARB antibiotic resistance. With the hope of achieving interdisciplinary one-health prevention and control of bacterial resistance.

Signal transducer and activator of transcription 3 (STAT3) is a transcriptional factor involved in almost all cancer hallmark features including tumor proliferation, metastasis, angiogenesis, immunosuppression, tumor inflammation, metabolism reprogramming, drug resistance, cancer stemness. Therefore, STAT3 has become a promising therapeutic target in a wide range of cancers. This review focuses on the up-to-date knowledge of STAT3 signaling in cancer. We summarize both the positive and negative modulators of STAT3 together with the cancer hallmarks involving activities regulated by STAT3 and highlight its extremely sophisticated regulation on immunosuppression in tumor microenvironment and metabolic reprogramming. Direct and indirect inhibitors of STAT3 in preclinical and clinical studies also have been summarized and discussed. Additionally, we highlight and propose new strategies of targeting STAT3 and STAT3-based combinations with established chemotherapy, targeted therapy, immunotherapy and combination therapy. These efforts may provide new perspectives for STAT3-based target therapy in cancer.

Evolving concepts on Parkinson's disease (PD) pathology suggest that α-synuclein (aSYN) promote dopaminergic neuron dysfunction and death through accumulating in the mitochondria. However, the consequence of mitochondrial aSYN localisation on mitochondrial structure and bioenergetic functions in neuronal cells are poorly understood. Therefore, we investigated deleterious effects of mitochondria-targeted aSYN in differentiated human dopaminergic neurons in comparison with wild-type (WT) aSYN overexpression and corresponding EGFP (enhanced green fluorescent protein)-expressing controls. Mitochondria-targeted aSYN enhanced mitochondrial reactive oxygen species (ROS) formation, reduced ATP levels and showed severely disrupted structure and function of the dendritic neural network, preceding neuronal death. Transmission electron microscopy illustrated distorted cristae and many fragmented mitochondria in response to WT-aSYN overexpression, and a complete loss of cristae structure and massively swollen mitochondria in neurons expressing mitochondria-targeted aSYN. Further, the analysis of mitochondrial bioenergetics in differentiated dopaminergic neurons, expressing WT or mitochondria-targeted aSYN, elicited a pronounced impairment of mitochondrial respiration. In a pharmacological compound screening, we found that the pan-caspase inhibitors QVD and zVAD-FMK, and a specific caspase-1 inhibitor significantly prevented aSYN-induced cell death. In addition, the caspase inhibitor QVD preserved mitochondrial function and neuronal network activity in the human dopaminergic neurons overexpressing aSYN. Overall, our findings indicated therapeutic effects by caspase-1 inhibition despite aSYN-mediated alterations in mitochondrial morphology and function.

Systemic lupus erythematosus (SLE) is responsive to treatment with immunosuppressives and steroids, but often pursues a relapsing or refractory course resulting in increasing incapacity and reduced survival. Autologous stem cell transplantation (ASCT) following immunoablative chemotherapy is a newer therapy for autoimmune disease of potential use in severe SLE. A retrospective registry survey was carried out by the European Blood and Marrow Transplant and European League Against Rheumatism (EBMT/EULAR) registry. Data was collected from 53 patients with SLE treated by ASCT in 23 centres. Disease duration before ASCT was 59 (2-155) months (median, range), 44 (83%) were female, and median age was 29 (9-52) years. At the time of ASCT a median of seven American College of Rheumatology (ACR) diagnostic criteria for SLE were present (range 2-10) and 33 (62%) had nephritis. Peripheral blood stem cells were mobilized with cyclophosphamide and granulocyte colony stimulating factor in 93% of cases. Ex vivo CD34 stem cell selection was performed in 42% of patients. Conditioning regimens employed cyclophosphamide in 84%, anti-thymocyte globulin in 76% and lymphoid irradiation in 22%. The mean duration of follow-up after ASCT was 26 (0-78) months. Remission of disease activity (SLEDAI < 3) was seen in 33/50 (66%; 95%CI 52-80) evaluable patients by six months, of which 10/31 (32%; 95%CI 15-50) subsequently relapsed after six (3-40) months. Relapse was associated with negative anti-double stranded DNA (anti-dsDNA) antibodies before ASCT (P = 0.007). There were 12 deaths after 1.5 (0-48) months, of which seven (12%; 95%CI 3-21) were related to the procedure. Mortality was associated with a longer disease course before ASCT (P = 0.036). In conclusion, this registry study demonstrates the efficacy of ASCT for remission induction of refractory SLE, although mortality appeared high. The safety of this procedure is likely to be improved by patient selection and choice of conditioning regimen. The return of disease activity in one-third of patients might be reduced by long-term immunosuppressive therapy post-ASCT.

BACKGROUND: Hypoxia is a key feature of breast cancer, which affects cancer development, metastasis and metabolism. Previous studies suggested that circular RNAs (circRNAs) could participate in cancer progression and hypoxia regulation. This study aimed to investigate the role of circRNA differentially expressed in normal cells and neoplasia domain containing 4C (circDENND4C) in breast cancer progression under hypoxia. METHODS: ) for experiments in vitro. The expression levels of circDENND4C, microRNA-200b (miR-200b) and miR-200c were measured by quantitative real-time polymerase chain reaction. Glycolysis was investigated by glucose consumption, lactate production and hexokinase II (HK2) protein level. Migration and invasion were evaluated via trans-well assay and protein levels of matrix metallopeptidase 9 (MMP9) and MMP2. The interaction between circDENND4C and miR-200b or miR-200c was explored by bioinformatics analysis, luciferase assay and RNA immunoprecipitation. Murine xenograft model was established to investigate the anti-cancer role of circDENND4C in vivo. RESULTS: circDENND4C highly expressed in breast cancer was up-regulated in response to hypoxia. Knockdown of circDENND4C decreased glycolysis, migration and invasion in breast cancer cells under hypoxia. circDENND4C was validated as a sponge of miR-200b and miR-200c. Deficiency of miR-200b or miR-200c reversed the suppressive effect of circDENND4C knockdown on breast cancer progression. Moreover, silence of circDENND4C reduced xenograft tumor growth by increasing miR-200b and miR-200c. CONCLUSION: circDENND4C silence suppresses glycolysis, migration and invasion in breast cancer cells under hypoxia by increasing miR-200b and miR-200c.

BACKGROUND: Inflammation is associated with perinatal brain injury but the underlying mechanisms are not completely characterized. Stimulation of Toll-like receptors (TLRs) through specific agonists induces inflammatory responses that trigger both innate and adaptive immune responses. The impact of engagement of TLR2 signaling pathways on the neonatal brain is still unclear. The aim of this study was to investigate the potential effect of a TLR2 agonist on neonatal brain development. METHODOLOGY/PRINCIPAL FINDINGS: Mice were injected intraperitoneally (i.p.) once a day from postnatal day (PND) 3 to PND11 with endotoxin-free saline, a TLR2 agonist Pam(3)CSK(4) (5 mg/kg) or Lipopolysaccharide (LPS, 0.3 mg/kg). Pups were sacrificed at PND12 or PND53 and brain, spleen and liver were collected and weighed. Brain sections were stained for brain injury markers. Long-term effects on memory function were assessed using the Trace Fear Conditioning test at PND50. After 9 days of Pam(3)CSK(4) administration, we found a decreased volume of cerebral gray matter, white matter in the forebrain and cerebellar molecular layer that was accompanied by an increase in spleen and liver weight at PND12. Such effects were not observed in Pam3CSK4-treated TLR 2-deficient mice. Pam3CSK4-treated mice also displayed decreased hippocampus neuronal density, and increased cerebral microglia density, while there was no effect on caspase-3 or general cell proliferation at PND12. Significantly elevated levels of IL-1β, IL-6, KC, and MCP-1 were detected after the first Pam3CSK4 injection in brain homogenates of PND3 mice. Pam(3)CSK(4) administration did not affect long-term memory function nor the volume of gray or white matter. CONCLUSIONS/SIGNIFICANCE: Repeated systemic exposure to the TLR2 agonist Pam(3)CSK(4) can have a short-term negative impact on the neonatal mouse brain.

The pathogenesis of stroke, trauma and chronic degenerative diseases, such as Alzheimer's disease (AD), has been linked to excitotoxic processes due to inappropriate stimulation of the N-methyl-D-aspartate receptor (NMDA-R). Attempts to use potent competitive NMDA-R antagonists as neuroprotectants have shown serious side-effects in patients. As an alternative approach, we were interested in the anti-excitotoxic properties of memantine, a well-tolerated low affinity uncompetitive NMDA-R antagonist presently used as an anti-dementia agent. We explored in a series of models of increasing complexity, whether this voltage-dependent channel blocker had neuroprotective properties at clinically relevant concentrations. As expected, memantine protected neurons in organotypic hippocampal slices or dissociated cultures from direct NMDA-induced excitotoxicity. However, low concentrations of memantine were also effective in neuronal (cortical neurons and cerebellar granule cells) stress models dependent on endogenous glutamate stimulation and mitochondrial stress, i.e. exposure to hypoxia, the mitochondrial toxin 1-methyl-4-phenylpyridinium (MPP+) or a nitric oxide (NO) donor. Furthermore, memantine reduced lethality and brain damage in vivo in a model of neonatal hypoxia-ischemia (HI). Finally, we investigated functional rescue (neuronal capacity to migrate along radial glia) by memantine in cerebellar microexplant cultures exposed to the indirect excitotoxin 3-nitropropionic acid (3-NP). Potent NMDA-R antagonists, such as (+)MK-801, are known to block neuronal migration in microexplant cultures. Interestingly, memantine significantly restored the number of neurons able to migrate out of the stressed microexplants. These findings suggest that inhibition of the NMDA-R by memantine is sufficient to block excitotoxicity, while still allowing some degree of signalling.