BMAL1 regulates mitochondrial fission and mitophagy through mitochondrial protein BNIP3 and is critical in the development of dilated cardiomyopathy

Dysregulation of circadian rhythms associates with cardiovascular disorders. It is known that deletion of the core circadian gene Bmal1 in mice causes dilated cardiomyopathy. However, the biological rhythm regulation system in mouse is very different from that of humans. Whether BMAL1 plays a role in regulating human heart function remains unclear. Here we generated a BMAL1 knockout human embryonic stem cell (hESC) model and further derived human BMAL1 deficient cardiomyocytes. We show that BMAL1 deficient hESC-derived cardiomyocytes exhibited typical phenotypes of dilated cardiomyopathy including attenuated contractility, calcium dysregulation, and disorganized myofilaments. In addition, mitochondrial fission and mitophagy were suppressed in BMAL1 deficient hESC-cardiomyocytes, which resulted in significantly attenuated mitochondrial oxidative phosphorylation and compromised cardiomyocyte function. We also found that BMAL1 binds to the E-box element in the promoter region of BNIP3 gene and specifically controls BNIP3 protein expression. BMAL1 knockout directly reduced BNIP3 protein level, causing compromised mitophagy and mitochondria dysfunction and thereby leading to compromised cardiomyocyte function. Our data indicated that the core circadian gene BMAL1 is critical for normal mitochondria activities and cardiac function. Circadian rhythm disruption may directly link to compromised heart function and dilated cardiomyopathy in humans.

BMAL1 regulates mitochondrial fission and mitophagy through mitochondrial protein BNIP3 and is critical in the development of dilated cardiomyopathy

Dysregulation of circadian rhythms associates with cardiovascular disorders. It is known that deletion of the core circadian gene Bmal1 in mice causes dilated cardiomyopathy. However, the biological rhythm regulation system in mouse is very different from that of humans. Whether BMAL1 plays a role in regulating human heart function remains unclear. Here we generated a BMAL1 knockout human embryonic stem cell (hESC) model and further derived human BMAL1 deficient cardiomyocytes. We show that BMAL1 deficient hESC-derived cardiomyocytes exhibited typical phenotypes of dilated cardiomyopathy including attenuated contractility, calcium dysregulation, and disorganized myofilaments. In addition, mitochondrial fission and mitophagy were suppressed in BMAL1 deficient hESC-cardiomyocytes, which resulted in significantly attenuated mitochondrial oxidative phosphorylation and compromised cardiomyocyte function. We also found that BMAL1 binds to the E-box element in the promoter region of BNIP3 gene and specifically controls BNIP3 protein expression. BMAL1 knockout directly reduced BNIP3 protein level, causing compromised mitophagy and mitochondria dysfunction and thereby leading to compromised cardiomyocyte function. Our data indicated that the core circadian gene BMAL1 is critical for normal mitochondria activities and cardiac function. Circadian rhythm disruption may directly link to compromised heart function and dilated cardiomyopathy in humans.

BMAL1 regulates mitochondrial fission and mitophagy through mitochondrial protein BNIP3 and is critical in the development of dilated cardiomyopathy

Dysregulation of circadian rhythms associates with cardiovascular disorders. It is known that deletion of the core circadian gene Bmal1 in mice causes dilated cardiomyopathy. However, the biological rhythm regulation system in mouse is very different from that of humans. Whether BMAL1 plays a role in regulating human heart function remains unclear. Here we generated a BMAL1 knockout human embryonic stem cell (hESC) model and further derived human BMAL1 deficient cardiomyocytes. We show that BMAL1 deficient hESC-derived cardiomyocytes exhibited typical phenotypes of dilated cardiomyopathy including attenuated contractility, calcium dysregulation, and disorganized myofilaments. In addition, mitochondrial fission and mitophagy were suppressed in BMAL1 deficient hESC-cardiomyocytes, which resulted in significantly attenuated mitochondrial oxidative phosphorylation and compromised cardiomyocyte function. We also found that BMAL1 binds to the E-box element in the promoter region of BNIP3 gene and specifically controls BNIP3 protein expression. BMAL1 knockout directly reduced BNIP3 protein level, causing compromised mitophagy and mitochondria dysfunction and thereby leading to compromised cardiomyocyte function. Our data indicated that the core circadian gene BMAL1 is critical for normal mitochondria activities and cardiac function. Circadian rhythm disruption may directly link to compromised heart function and dilated cardiomyopathy in humans.

Quantitative structure-activity relationship prediction of carcinogenicity of N-nitroso compounds based on category approach and read-across / 中华预防医学杂志

Objective@#New quantitative structure-activity relationship (QSAR) method was used to predict N-nitroso compounds (NOCs) carcinogenicity. This could provide evidences for health risk assessment of the chemicals.@*Methods@#Total 74 chemical substances of NOCs were included as target chemicals for this validation study by using QSAR Toolbox based on category approach and read-across. The included 74 NOCs were categorized and subcategorized respectively using "Organic functional groups, Norbert Haider " profiler and "DNA binding by OASIS V.1.1" profiler. Carcinogenicity of rat were used as target of prediction, the carcinogenicity@*results@#of analogues in chemical categories were cross-read to obtain the carcinogenic predictive results of the target chemicals. Results 74 NOCs included 26 nonclic N-nitrosamines, 24 cyclic N-nitrosamines and 24 N-nitrosamides The sensitivity, specificity and concordance of the category approach and read-across for predicting carcinogenicity of 74 NOCs were 75% (48/64), 70%(7/10) and 74% (55/74) respectively. The concordance for noncyclic N-nitrosamines, cyclic N-nitrosamines and N-nitrosamides were 88% (23/26), 71% (17/24) and 63% (15/24) respectively.@*Conclusion@#QSAR based on category approach and read-across is good for prediction of NOCs carcinogenicity, and can be used for high-throughput qualitative prediction of NOCs carcinogenicity.

Drug Discovery Investigation of Nourishing Kidney-yin And Warming Kidney-yang Granules in Prevention And Treatment of Primary Osteoporosis Based on Theory of "Treating Both Disease And Traditional Chinese Medicine Syndrome" / 世界科学技术-中医药现代化

Nourishing kidney-yin (NKY) granules and warming kidney-yang (WKY) granules represent one of the prescriptions that prescribed in treating primary osteoporosis (POP) in light of tonifying kidney and nourishing essence principle as well as the theory of "treating both the disease and traditional Chinese medicine (TCM) syndrome".Both granules were created through the systematic analysis of clinic prescriptions by Professor Shi Qi.Consequently clinical investigations have well established that NKY granules significant improved bone mineral density (BMD) as well as relieved the kidney-yin deficiency syndromes in POP patients.Meanwhile,WKY granules relieve kidney-yang deficiency syndrome and the quality of life (QOL).What is more,pharmacological study established the application of common cnidium fruit,and fructus ligustri lucidi alleviated bone loss in OVX-induced mice.In addition,investigation with effective components identified that both NKY and WKY granules play systematic pharmacological effects on bone remodeling by regulating the expression of BMP/Smad,Wnt/β-catenin,RANKL/RANK/OPG axis,and Notch.The drug discovery was performed by the lead of traditional Chinese medicine (TCM) theory.It is one successful transformation investigation based on pharmacological effects,clinical intervention,animal model,cell culture and molecular investigation.

Clinical curative effects of mifepristone in treatment of perimenopausal dysfunctional uterine bleeding / 中国基层医药

Objective To investigate the clinical effect of mifepristone in treatment of perimenopausal dysfunctional uterine bleeding.Methods 80 cases of patients with perimenopausal dysfunctional uterine bleeding were selected and randomly divided into control group and research group,with 40 cases in each group.The control group was in the diagnosis of curettage after 10 days of oral provera 10mg every night before going to bed,stopped drug application in 14 days,15 days to retreat hemorrhages oral provera 10mg/day,stayed for 10 days,repeated drug use 5 months.The research group was treated with oral mifepristone every night before going to bed since cleaning up the uterus,12.5mg,for 6 months.Outpatient follow -up for 6 months,the recent curative effect,long -term curative effect,hormone level changes,anemia and adverse drug reactions were observed.Results The total effective rate of the research group was 95.0%,which of the control group was 82.5%,there was statistically significant difference(χ2 =7.82, P =0.005);The recurrence rate of the research group was 5.0%,which of the control group was 15.0%,there was statistically significant difference(χ2 =5.56,P =0.018).The FSH,LH,E2 and P of the two groups were decreased, but which of the research group were lower significantly than those of control group,the differences were statistically significant (all P <0.05).The decreased degree of endometrium in the research group was larger than that of the control group,the difference was statistically significant(P =0.0001).Conclusion Treatment of mifepristone perimenopausal dysfunctional uterine bleeding is significant,safe and convenient,which is worthy of clinical application.

The roles of Mesp family proteins: functional diversity and redundancy in differentiation of pluripotent stem cells and mammalian mesodermal development

Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies.

Inhibitory activities of 3-O-β-chacotriosyl pentacyclic triterpenoids against the entry of H5 N1 influenza viruses in vitro / 中国药理学通报

Aim To study the inhibitory activities of potential new anti-influenza virus agents,3-O-β-chaco-triosyl pentacyclic triterpenoids against the entry of H5N1influenza viruses.Methods Three target com-pounds were designed and synthesized structurally re-lated to the lead compound 3-O-β-chacotriosyl dioscin derivative (1 )with inhibitory activities against H5N1 influenza viruses.The inhibitory activities of these tar-get compounds were tested at a cellular level pseudo vi-rus system targeting H5N1 influenza viruse entry.Re-sults All the compounds 1 a,1 b and 1 c showed po-tent inhibitory activities against the entry of A/Thai-land/Kan353/2004 pseudo virus into the target cells, of which compound 1 b showed the best inhibitory activ-ity with an IC50 value of (1.25 ±0.22)μmol·L-1. Conclusion The SARs analysis of these compounds indicated that replacement of the aglycone moiety of compound 1 with pentacyclic triterpenoids could in-crease antiviral activity.Different types of pentacyclic triterpen as aglycone residue had the significant influ-ence on the inhibitory activity (1 b >1 c >1 a),sug-gesting ursane type of triterpenes was superior to the two other kinds of triterpenes as aglycone residue.

Effect of Er:YAG laser on the dynamic changes in subgingival flora in type 2 diabetic patients with periodontitis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the changes in subgingival microflora before and after Er:YAG laser treatment on diabetic patients with periodontitis, and to compare with the subgingival microflora of chronic periodontitis.</p><p><b>METHODS</b>Subgingival plaque of 13 pairs of teeth (26 sites) was selected from type 2 diabetic patients at pretreatment, one month post-treatment, and three months post-treatment. Subgingival plaque was also obtained from 11 cases of moderate to severe chronic periodontitis with similar severity of periodontitis. The DNA of the subgingival plaque samples was extracted. Whole bacterial 16S rDNA gene fragments separated by denaturing gradient gel electrophoresis. Specific DNA bands were then chosen for retrieval and sequencing.</p><p><b>RESULTS</b>The gene sequencing results of the special DNA bands of subgingival plaque samples show that the pathogenic bacteria of both diabetic periodontitis and simple chronic periodontitis were Prevotella intermedia and Tannerella forsythia, respectively. The composition of the subgingival microflora before and after laser treatment changed. Some DNA bands, including that of Tannerella forsythia, disappeared or weakened one month after treatment. A new strip appeared, which belonged to Actinomyces sp.</p><p><b>CONCLUSION</b>The profiles of the subgingival microflora changed after treatment, and one month was indicated as an important stage. Er:YAG laser may have an important function in delaying microflora recolonization.</p>

Mechanism of resveratrol on the promotion of induced pluripotent stem cells / 中西医结合学报

To investigate the effects of resveratrol (RV) in reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPSCs) and the related mechanism.

Expression of BMP4 mature peptide in eukaryotic cells and its differentiation-inhibiting effect in culturing induced pluripotent stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of bone morphogenetic protein 4 (BMP4) in culturing induced pluripotent stem cells (iPSCs) and the related signal pathways.</p><p><b>METHODS</b>We amplified the mature peptide of BMP4 from the placenta through RT-PCR, and IgK secretion peptide was ligated to the N-terminal of BMP4 mature peptide. The recombinant plasmid pPYCAG-IgK-BMP4 was transfected into 293T cells and screened with puromycin, and the positive clones for expressing BMP4 were verified by cell immunofluorescence and Western blotting. To test the bioactivity of BMP4, iPSCs were cultured in the medium supplemented with leukemia inhibitory factor (LIF) plus the supernatant containing BMP4, and the cell phenotype, cell differentiation capacity into lineages of the 3 germ layers and expression levels of pluripotency-associated genes were investigated.</p><p><b>RESULTS</b>Smad1 was phosphorylated by BMP4 from the culture medium. iPSCs cultured in the medium supplemented with LIF plus the supernatant containing BMP4 for 3 passages maintained the phenotype of stem cells with the expression levels of pluripotency-associated genes not affected. These iPSCs also maintained the capacity to differentiate into cell lineages of the 3 germ layers.</p><p><b>CONCLUSION</b>BMP4 can be efficiently expressed in mammalian cells to maintain the multipotent differentiation capacity of the iPSCs in in vitro culture.</p>

Effects of Qishe Pill on vertebral hyperostosis induced by upright posture in rats / 中西医结合学报

To observe the effects of Qishe Pill, a compound traditional Chinese herbal medicine, on lumbar vertebral bone formation induced by long-time upright posture in rats and to investigate the potential mechanism.

Preventive and therapeutic effects of Yiqi Huayu Recipe on degeneration of articular cartilage in rats with osteoarthritis / 中西医结合学报

To study the efficacy and possible mechanism of Yiqi Huayu Recipe (YQHYR), a compound traditional Chinese herbal medicine, in preventing and treating degeneration of the articular cartilage in rats with osteoarthritis (OA).

Mechanism of Yiqi Huayu Bushen recipe in treating cervical syndrome in rats with qi deficiency, blood stasis and kidney deficiency / 中西医结合学报

To explore the mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound traditional Chinese herbal medicine, in treating cervical syndrome (CS) with qi deficiency, blood stasis and kidney deficiency in rats.

Mechanism of Yiqi Huayu Bushen Recipe in treating cervical syndrome with kidney deficiency in rats / 中西医结合学报

To research the action mechanism of Yiqi Huayu Bushen Recipe (YHBR), a compound of traditional Chinese herbal medicine, in treating cervical syndrome (CS) with kidney deficiency in rats.

Establishment of a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency / 中西医结合学报

To establish a rat model of cervical syndrome with qi deficiency, blood stasis and kidney deficiency.

Establishment of a rat model of cervical syndrome with kidney deficiency / 中西医结合学报

OBJECTIVE: To establish a rat model of cervical syndrome (CS) with kidney deficiency. METHODS: A group of 30 three-month-old female Sprague-Dawley rats were randomly divided into normal control group, CS group and CS with kidney deficiency group (combined group), with 10 rats in each group. Rats in the normal control group received no treatment, rats in the CS group underwent only resection of cervical muscles and ligaments as unbalanced dynamic and static animal model, and rats in combined group underwent resection of both cervical muscles and ovaries as kidney deficiency model. Serum and cervical intervertebral discs were collected. Kidney deficiency was determined by observing the morphologic changes of uterus and appendages, detecting the weight of uterus and appendages and the content of serum estradiol (E(2)). The degeneration of intervertebral discs was determined by detecting the histopathology, the expressions of type II collagen and type X collagen proteins, and the expressions of aggrecan-1 (Agc1), type II procollagen gene (Col2a1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNAs. RESULTS: Compared with those in the normal control group and CS group, the rats in the combined group were noted with the uterus atrophied, the caliber of oviduct thinned, the weight of uterus and appendages diminished obviously, the content of serum E(2) decreased, cervical intervertebral disc degenerated more seriously, type II collagen protein expression decreased, type X collagen protein expression increased, Agc1 and Col2a1 mRNA expressions in intervertebral disc decreased, and the MMP-13 mRNA expression increased. CONCLUSION: The rat model of CS with kidney deficiency is established. Kidney deficiency can aggravate cervical intervertebral disc degeneration.

Construction of RNAi Expression Vector by Fusion Gene Fragments of BBE and COR from Papaver nudicarule / 中国生物工程杂志

The codeinone reductase gene (cor) and the berberine bridge enzyme gene (bbe) were cloned from young leaf of opium poppy(Papaver somniferum L) by RT-PCR and the coding sequences of these gene were analyzed. The result demonstrated that the cloned COR gene sequence was highly homologous to the other COR gene family members showing 98.96% identity to COR1.1. The cloned BBE gene sequence was 94.84% identified with the released BBE genes in GenBank previously. Based on the cDNA sequences of COR and BBE,two fragments about 400～500 bp from each gene with lower identity among them were cloned. The fusion gene BC(744 bp) is fused by the PCR technique. Then the promoter CaMV 35S driven,containing'forward BC fusion fragments-reverse pdk intron-reverse BC fusion fragments',plant siRNA expression vector were constructed based on the vectors pHANNIBAL and pART27. Inhibition efficiency of the expression vector to the morphine synthesis was studied by transforming papaver nudicarule.The work will lay the foundation for breeding a low morphine and high thebaine poppy.