Tripeptidyl peptidase II coordinates the homeostasis of calcium and lipids in the central nervous system and its depletion causes presenile dementia in female mice through calcium/lipid dyshomeostasis-induced autophagic degradation of CYP19A1.

Rationale: Tripeptidyl peptidase II (TPP2) has been proven to be related to human immune and neurological diseases. It is generally considered as a cytosolic protein which forms the largest known protease complex in eukaryotic cells to operate mostly downstream of proteasomes for degradation of longer peptides. However, this canonical function of TPP2 cannot explain its role in a wide variety of biological and pathogenic processes. The mechanistic interrelationships and hierarchical order of these processes have yet to be clarified. Methods: Animals, cells, plasmids, and viruses established and/or used in this study include: TPP2 knockout mouse line, TPP2 conditional knockout mouse lines (different neural cell type oriented), TRE-TPP2 knockin mouse line on the C57BL/6 background; 293T cells with depletion of TPP2, ATF6, IRE1, PERK, SYVN1, UCHL1, ATG5, CEPT1, or CCTα, respectively; 293T cells stably expressing TPP2, TPP2 S449A, TPP2 S449T, or CCTα-KDEL proteins on the TPP2-depleted background; Plasmids for eukaryotic transient expression of rat CYP19A1-Flag, CYP19A1 S118A-Flag, CYP19A1 S118D-Flag, Sac I ML GFP Strand 11 Long, OMMGFP 1-10, G-CEPIA1er, GCAMP2, CEPIA3mt, ACC-GFP, or SERCA1-GFP; AAV2 carrying the expression cassette of mouse CYP19A1-3 X Flag-T2A-ZsGreen. Techniques used in this study include: Flow cytometry, Immunofluorescence (IF) staining, Immunohistochemical (IHC) staining, Luxol fast blue (LFB) staining, ß-galactosidase staining, Lipid droplet (LD) staining, Calcium (Ca2+) staining, Stimulated emission depletion (STED) imaging, Transmission electron microscopic imaging, Two-photon imaging, Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end Labeling (TUNEL) assay, Bromodeoxyuridine (BrdU) assay, Enzymatic activity assay, Proximity ligation assay (PLA), In vivo electrophysiological recording, Long-term potentiation (LTP) recording, Split-GFP-based mitochondria-associated membrane (MAM) detection, Immunoprecipitation (IP), Cellular fractionation, In situ hybridization, Semi-quantitative RT-PCR, Immunoblot, Mass spectrometry-based lipidomics, metabolomics, proteomics, Primary hippocampal neuron culture and Morris water maze (MWM) test. Results: We found that TPP2, independent of its enzymatic activity, plays a crucial role in maintaining the homeostasis of intracellular Ca2+ and phosphatidylcholine (PC) in the central nervous system (CNS) of mice. In consistence with the critical importance of Ca2+ and PC in the CNS, TPP2 gene ablation causes presenile dementia in female mice, which is closely associated with Ca2+/PC dysregulation-induced endoplasmic reticulum (ER) stress, abnormal autophagic degradation of CYP19A1 (aromatase), and estrogen depletion. This work therefore uncovers a new role of TPP2 in lipogenesis and neurosteroidogenesis which is tightly related to cognitive function of adult female mice. Conclusion: Our study reveals a crucial role of TPP2 in controlling homeostasis of Ca2+ and lipids in CNS, and its deficiency causes sexual dimorphism in dementia. Thus, this study is not only of great significance for elucidating the pathogenesis of dementia and its futural treatment, but also for interpreting the role of TPP2 in other systems and their related disorders.

Synotisjinpingensis (Asteraceae, Senecioneae), a new species with white ray florets from southeastern Yunnan, China.

Synotisjinpingensis (Asteraceae, Senecioneae), a new species from Jinping county in southeastern Yunnan province, China, is described and illustrated. This species is distinguished by having white ray florets in the genus Synotis, in which only species with yellow ray florets have been hitherto known. In habit and leaf shape S.jinpingensis is most closely similar to S.duclouxii, a species occurring in southwestern Guizhou, southern Sichuan and northeastern Yunnan, China, but differs, in addition to the color of ray florets, by having fewer lateral veins of leaves, obviously longer bracts of calyculus, and larger phyllaries. The membership of the new species within Synotis is strongly corroborated by evidence from floral micromorphology and phylogenetic analyses based on ITS sequence data. Color photographs of living plants, a distribution map, and provisional IUCN status of S.jinpingensis are provided.

Ultra-centrifugation force in adaptive evolution changes the cell structure of oleaginous yeast Trichosporon cutaneum into a favorable space for lipid accumulation.

Microbial lipid production from lignocellulose biomass provides an essential option for sustainable and carbon-neutral supply of future aviation fuels, biodiesel, as well as various food and nutrition products. Oleaginous yeast is the major microbial cell factory but its lipid-producing performance is far below the requirements of industrial application. Here we show an ultra-centrifugation fractionation in adaptive evolution (UCF) of Trichosporon cutaneum based on the minor cell density difference. The lightest cells with the maximum intracellular lipid content were isolated by ultra-centrifugation fractionation in the long-term adaptive evolution. Significant changes occurred in the cell morphology with a fragile cell wall wrapping and enlarged intracellular space (two orders of magnitude increase in cell size). Complete and coordinate assimilations of all nonglucose sugars derived from lignocellulose were triggered and fluxed into lipid synthesis. Genome mutations and significant transcriptional regulations of the genes responsible for cell structure were identified and experimentally confirmed. The obtained T. cutaneum MP11 cells achieved a high lipid production of wheat straw, approximately five-fold greater than that of the parental cells. The study provided an effective method for screening the high lipid-containing oleaginous yeast cells as well as the intracellular products accumulating cells in general.

Molecular signaling pathways in the occurrence and development of osteoarthritis / 中国组织工程研究

BACKGROUND: Osteoarthritis is a refractory degenerative disease that seriously affects patients’ quality of life, and brings a heavy financial burden to their families. Nowadays, treatments for osteoarthritis can only alleviate symptoms, improve joint function in a short period of time, but cannot prevent or reverse the pathological process of osteoarthritis. To date, there are no prophylactic or regenerative therapies for osteoarthritis. The signaling pathway can explain the occurrence and development of disease at the molecular level. Therefore, the regulation of signaling pathway can be used as a potential treatment for osteoarthritis. OBJECTIVE: To review the occurrence and development of osteoarthritis, aiming at providing new ideas for clinical diagnosis, treatment and deeper understanding of diseases in the future. METHODS: CNKI, Wanfang, VlP, CBMdisc, PubMed and MEDLINE databases were retrieved respectively with the keywords of “osteoarthritis; signaling pathway; EGFR; Hedgehog; NF-kB; Notch” in Chinese and English, and included literature reports regarding the signaling pathways related to osteoarthritis published in 2008-2019 were retrieved. RESULTS AND CONCLUSION: Normal articular cartilage function depends on the regulation of multiple signaling pathways, and the occurrence and development of osteoarthritis are also regulated by corresponding signaling pathways, providing deep understanding of osteoarthritis at the molecular level and new ideas for clinical diagnosis and treatment. Regulation of signaling pathways through drug action and gene modification is a potential treatment for osteoarthritis. However, there is a common problem in related research: most of the research on signaling pathway is isolated, and the occurrence and development of osteoarthritis may involve the joint action of multiple signal pathways. Therefore, further research is needed on the synergy of signaling pathways.

Effects of Acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) on Behaviors and Monoamine Neurotransmitter in Prefrontal Cortex of Attention Deficit Hyperactivity Disorder Rats / 中国中医药信息杂志

Objective To observe the effects of acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) on behaviors and contents of monoamine neurotransmitter dopamine (DA), noradrenalin (NE) and 5-hydroxytrypamine (5-HT) in prefrontal cortex (PFC) of attention deficit hyperactivity disorder (ADHD) rats; To discuss mechanism of action of acupuncture for prevention and treatment of ADHD. Methods 4-week-old SHR rats were randomly divided into the model group, Western medicine group and acupuncture group, and WKY rats with the same age were selected as the normal control group, with 10 rats in each group. Acupuncture was applied to "Sanyinjiao" (SP6) and "Baihui" (GV20) of rats in acupuncture group for 15 min. The rats in Western medicine group was given MPH (ritalin) for gavage, and rats were treated for 4 weeks in the dark-phase. Open field test (OFT), elevated plus maze (EPM) and novel object recognition test (NORT) were conducted to evaluate the spontaneous activity, impulsivity and learning-memorial ability of rats individually at the end of treatment. Contents of DA, NE and 5-HT in PFC were detected by HPLC. Results Compared with the normal control group, all of the behavior parameters of rats in model group were significantly higher (P＜0.05), and the contents of DA, NE and 5-HT in PFC of model group were lower than those of normal control group (P＜0.05). Compared with the model group, acupuncture group showed shorter movement distance, less rearing and fewer grooming activities in the OFT; percentages of times of entering the open arms and staying duration were reduced in the total period in the EPM; preference index in the NORT was elevated (P＜0.05); the contents of NE and 5-HT in PFC of acupuncture group increased significantly (P＜0.05). Conclusion Acupuncture at "Sanyinjiao" (SP6) and "Baihui" (GV20) canreduce the spontaneous activity and impulsivity, and improve learning and memory ability of ADHD model rats, which may be related to elevated contents of NE and 5-HT in the PFC.

Vatalanib, a tyrosine kinase inhibitor, decreases hepatic fibrosis and sinusoidal capillarization in CCl4-induced fibrotic mice.

Among the various consequence arising from lung injury, hepatic fibrosis is the most severe. Decreasing the effects of hepatic fibrosis remains one of the primary therapeutic challenges in hepatology. Dysfunction of hepatic sinusoidal endothelial cells is considered to be one of the initial events that occur in liver injury. Vascular endothelial growth factor signaling is involved in the progression of genotype changes. The aim of the present study was to determine the effect of the tyrosine kinase inhibitor, vatalanib, on hepatic fibrosis and hepatic sinusoidal capillarization in a carbon tetrachloride (CCl4)­induced mouse model of liver fibrosis. Liver fibrosis was induced in BALB/c mice using CCl4 by intraperitoneal injection for 6 weeks. The four experimental groups included a control, and three experimental groups involving administration of CCl4, vatalanib and a combination of the two. Histopathological staining and measuring live hydroxyproline content evaluated the extent of liver fibrosis. The expression of α­smooth muscle actin (SMA) and cluster of differentiation (CD) 34 was detected by immunohistochemistry. Collagen type I, α­SMA, transforming growth factor (TGF)­ß1 and vascular endothelial growth factor receptor (VEGFR) expression levels were measured by reverse transcription-quantitative polymerase chain reaction (RT­qPCR). The average number of fenestrae per hepatic sinusoid was determined using transmission electron microscopy. Liver fibrosis scores and hydroxyproline content were decreased in both vatalanib groups. In addition, both doses of vatalanib decreased mRNA expression levels of hepatic α-SMA, TGF-ß1, collagen­1, VEGFR1, and VEGFR2. Levels of α­SMA and CD34 protein were decreased in the vatalanib group compared with the CCl4 group. There were significant differences in the number of fenestrae per sinusoid between the groups. The present study identified that administration of vatalanib was associated with decreased liver fibrosis and hepatic sinusoidal capillarization in CCl4-induced mouse models, and is a potential compound for counteracting liver fibrosis.