37 results on '"Lida Guo"'
Search Results
2. Vitamin C prevents hypogonadal bone loss.
- Author
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Ling-Ling Zhu, Jay Cao, Merry Sun, Tony Yuen, Raymond Zhou, Jianhua Li, Yuanzhen Peng, Surinder S Moonga, Lida Guo, Jeffrey I Mechanick, Jameel Iqbal, Liu Peng, Harry C Blair, Zhuan Bian, and Mone Zaidi
- Subjects
Medicine ,Science - Abstract
Epidemiologic studies correlate low vitamin C intake with bone loss. The genetic deletion of enzymes involved in de novo vitamin C synthesis in mice, likewise, causes severe osteoporosis. However, very few studies have evaluated a protective role of this dietary supplement on the skeleton. Here, we show that the ingestion of vitamin C prevents the low-turnover bone loss following ovariectomy in mice. We show that this prevention in areal bone mineral density and micro-CT parameters results from the stimulation of bone formation, demonstrable in vivo by histomorphometry, bone marker measurements, and quantitative PCR. Notably, the reductions in the bone formation rate, plasma osteocalcin levels, and ex vivo osteoblast gene expression 8 weeks post-ovariectomy are all returned to levels of sham-operated controls. The study establishes vitamin C as a skeletal anabolic agent.
- Published
- 2012
- Full Text
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3. Optimization of ultrasonic assisted ethanol extraction of polyphenols from Phyllanthus emblica by Response Surface Methodology
- Author
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Yali Wu, Wei Zhang, Lida Guo, Suqing Zhang, and Shizhong Liang
- Abstract
Polyphenols are indispensable as the main antioxidant substances of P. embaldii. This experiment applied ultrasonic assisted the emblic leafflower fruit polyphenol was extracted with ethanol, and using the response surface method to optimize the extraction results improved, to obtain the parameters of the excellent conditions, mainly for the drying temperature of 80 ℃ emblic leafflower fruit pulp, 60% of the alcohol concentration, solid-liquid ratio, ultrasonic extraction time is 24 min and all the sons of required temperature 45 ℃ of ultrasound. Under the experimental conditions, the yield of polyphenols obtained from P. emballiae was 10.40%, which was almost the same as the ideal prediction value of 10.39%. The established regression model has a good fitting degree.
- Published
- 2023
4. Preclinical characterization of ABI-H2158, an HBV core inhibitor with dual mechanisms of action
- Author
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Ran Yan, Dawei Cai, Yuhua Zong, Lida Guo, Yi Zhou, Ariel Tang, Lichun Li, Qi Huang, Richard Colonno, and Michael A. Walker
- Subjects
Pharmacology ,Virology - Abstract
The HBV core protein plays an integral role in multiple steps of the HBV lifecycle. Consequently, HBV core inhibitors interrupt multiple steps of the replication cycle, including blocking pgRNA encapsidation and prematurely disassembling existing nucleocapsids, thereby preventing them from transporting relaxed circular (rcDNA) to the nucleus for conversion to covalently closed circular DNA (cccDNA). ABI-H2158 is an HBV core inhibitor that advanced into Phase 2 clinical trials for the treatment of chronic hepatitis B virus infection (cHBV) but was discontinued due to hepatotoxicity. Here, the potency, selectivity, and mechanisms of action of ABI-H2158 were evaluated using a variety of cell-based assays. Antiviral activity was measured by quantifying intracellular or secreted HBV DNA, RNA, and antigens. ABI-H2158 inhibited HBV replication by blocking pgRNA encapsidation in induced HepAD38 cells (EC
- Published
- 2022
5. Bacterial Diversity in Rhipicephalus sanguineus (Acari: Ixodidae) from Two States in Nigeria
- Author
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Jingze Liu, Hongyuan Zheng, Zhijun Yu, Lida Guo, Desmond O. Agwunobi, and Joshua Kamani
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0301 basic medicine ,biology ,Rhipicephalus sanguineus ,media_common.quotation_subject ,030106 microbiology ,Zoology ,biology.organism_classification ,03 medical and health sciences ,030104 developmental biology ,Insect Science ,Acari ,Agronomy and Crop Science ,Ecology, Evolution, Behavior and Systematics ,Ixodidae ,Diversity (politics) ,media_common - Abstract
Rhipicephalus sanguineus (Latreille) is the most widely distributed tick species globally and plays a major role in tick-borne pathogen transmission among canine populations. The microbial community of this tick has not been characterized previously in Nigeria. Thus, in this study, the bacterial diversity in R. sanguineus collected from two states in Nigeria was explored using 16S rRNA high-throughput sequencing (IonS5TM XL sequencing platform). Three hundred genera (although 18 groups belong to unidentified genera) of bacteria belonging to 22 phyla were detected after pooling and sequencing, indicating a diverse bacterial community profile. At the phylum level, Actinobacteria (47.39%) was the most abundant phylum in the Benue state samples, followed by Proteobacteria (43.87%) and Firmicutes (8.21%), whereas Fusobacteria (38.14%) was the most abundant phylum in Plateau state samples, followed by Bacteroidetes (17.57%) and Firmicutes (17.54%). Proteobacteria accounts for 17% of samples from Plateau state. In general, Cetobacterium (35.86%) was the most abundant genus, followed by an unidentified genus under Corynebacteriaceae (29.94%) and Stenotrophomonas (19.52%). Coxiella spp., one common endosymbiont found in various tick species, was present in R. sanguineus. This study provides a baseline of knowledge of the bacterial microbiome of R. sanguineus in Nigeria. Nonpathogenic bacteria species were prevalent, and not much is known about their role. Thus, their composition, functional, and ecologic implications merit further investigation because this will aid in the subsequent management of ticks and tick-borne diseases.
- Published
- 2021
6. Preclinical Profile and Characterization of the Hepatitis B Virus Core Protein Inhibitor ABI-H0731
- Author
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Kirk Henne, Yuhua Zong, Yi Zhou, Dawei Cai, Lida Guo, Ran Yan, Richard J. Colonno, Lichun Li, Alexandre Mercier, Ariel Tang, and Qi Huang
- Subjects
Hepatitis B virus ,pgRNA packaging ,medicine.disease_cause ,Virus Replication ,Antiviral Agents ,03 medical and health sciences ,Hepatitis B, Chronic ,Cp ,core protein ,medicine ,capsid ,Humans ,Pharmacology (medical) ,core inhibitor ,030304 developmental biology ,Pharmacology ,0303 health sciences ,030306 microbiology ,Chemistry ,Viral Core Proteins ,DNA replication ,cccDNA ,Entecavir ,Hepatitis C, Chronic ,Hepatitis B ,Virology ,body regions ,Infectious Diseases ,Capsid ,Mechanism of action ,Cell culture ,DNA, Viral ,medicine.symptom ,DNA, Circular ,pregenomic RNA ,Nucleoside ,medicine.drug - Abstract
ABI-H0731, a first-generation hepatitis B virus (HBV) core protein inhibitor, has demonstrated effective antiviral activity in chronic hepatitis B (CHB) patients in a phase 1b clinical trial and is currently being further evaluated in phase 2 clinical trials. Here, we report the preclinical profile of ABI-H0731. In in vitro cell culture systems (HepG2-derived cell lines HepAD38 and HepG2-NTCP and primary human hepatocytes [PHHs]), ABI-H0731 exhibited selective inhibition of HBV DNA replication (50% effective concentration [EC50] from 173 nM to 307 nM)., ABI-H0731, a first-generation hepatitis B virus (HBV) core protein inhibitor, has demonstrated effective antiviral activity in chronic hepatitis B (CHB) patients in a phase 1b clinical trial and is currently being further evaluated in phase 2 clinical trials. Here, we report the preclinical profile of ABI-H0731. In in vitro cell culture systems (HepG2-derived cell lines HepAD38 and HepG2-NTCP and primary human hepatocytes [PHHs]), ABI-H0731 exhibited selective inhibition of HBV DNA replication (50% effective concentration [EC50] from 173 nM to 307 nM). Most importantly, ABI-H0731 suppressed covalently closed circular DNA (cccDNA) formation in two de novo infection models with EC50s from 1.84 μM to 7.3 μM. Mechanism-of-action studies indicated that ABI-H0731 is a direct-acting antiviral that targets HBV core protein, preventing HBV pregenomic RNA (pgRNA) encapsidation and subsequent DNA replication. The combination of ABI-H0731 with entecavir appears to decrease viral DNA faster and deeper than nucleoside/nucleotide analogue (NrtI) therapy alone. In addition, ABI-H0731 disrupts incoming nucleocapsids, causing the premature release of relaxed circular DNA (rcDNA) before delivery to the nucleus, and thus prevents new cccDNA formation. ABI-H0731 exhibits pangenotypic activity and is additive to moderately synergistic when combined with an NrtI. In addition to its potency and novel mechanism of action, ABI-H0731 possesses drug-like properties and a preclinical pharmacokinetic profile supportive of once-daily dosing in patients with CHB. Taken together, these data support the ongoing clinical development of ABI-H0731 as a treatment for HBV.
- Published
- 2020
7. Density-controlled growth and passivation of ZnO nanorod arrays by electrodeposition
- Author
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Jie Chen, Linge Ma, Lida Guo, Fu-Kuo Chiang, and Yang Tang
- Subjects
Materials science ,Passivation ,Scanning electron microscope ,Doping ,Metals and Alloys ,Nanotechnology ,Crystal growth ,02 engineering and technology ,Surfaces and Interfaces ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Nanomaterials ,Chemical engineering ,Transmission electron microscopy ,Materials Chemistry ,Nanorod ,Thin film ,0210 nano-technology - Abstract
Here we established a simple electrochemical method of synthesizing ZnO nanomaterials with high quality and the ability to control the nanostructures' density For ZnO nanorod growth, a thin layer of Al doped ZnO (AZO) was deposited on glass substrates and ZnO nanorod arrays were grown on these substrates from aqueous solution containing Zn(NO 3 ) 2 and NH 4 NO 3 . It was found that the density, morphology, and quality of the nanorod arrays can be controlled by changing the applied potentials or the concentration of NH 4 NO 3 in the solution as growing and etching were carried out simultaneously. Moreover, the nanorod density decrease resulted in an improvement of the quality of the as-grown nanorods. Scanning electron microscope (SEM), transmission electron microscopy (TEM), photoluminescence and X-ray diffraction (XRD) have been used to characterize the samples. The density of ZnO nanorods can be controlled by varying the concentrations of the precursor solutions or applied potentials. Passivated low-density ZnO nanorod arrays with high quality were fabricated.
- Published
- 2017
8. Curcumin Combined with Oxaliplatin Effectively Suppress Colorectal Carcinomain vivoThrough Inducing Apoptosis
- Author
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Zhijun Yu, Duo Wang, Limeng Liu, Lida Guo, Jingze Liu, Li-jian Guo, Yong-qing Shen, and Xiao-han Zhao
- Subjects
Pharmacology ,Cell cycle checkpoint ,business.industry ,Colorectal cancer ,Poly ADP ribose polymerase ,medicine.disease ,Oxaliplatin ,chemistry.chemical_compound ,chemistry ,In vivo ,Apoptosis ,Survivin ,Curcumin ,medicine ,business ,medicine.drug - Abstract
Studies have shown chemopreventive and/or chemotherapeutic effects of several curcumin-based combinatorial treatments on colorectal cancer cells. However, their in vivo effects remain unclear. This study has demonstrated the therapeutic effect of curcumin and oxaliplatin, alone or in combination, on subcutaneously xenografted LoVo human colorectal cancer cells in immunodeficient (nu/nu) mice in vivo. Combinatorial administration of curcumin and oxaliplatin evidently inhibited the growth of colorectal cancer in nude mice, which was significantly more effective than either agent alone. Curcumin combined with oxaliplatin treatment induced apoptosis, accompanied by ultrastructural changes and cell cycle arrest in S and G2/M phases. Further mechanism analysis indicated that while the number of apoptotic tumor cells and the expression of Bax, caspase-3, and poly (ADP-ribose) polymerase (PARP) increased significantly, the expression of Bcl-2, survivin, HSP70, pro-caspase-3, and pro-PARP were dramatically suppressed in tumor cells after the treatment with combinatorial curcumin and oxaliplatin for 22 days. Taken together, the present study has demonstrated that administration of combined curcumin and oxaliplatin effectively suppressed colorectal carcinoma in vivo through inducing apoptosis and thus may provide an effective treatment for colorectal carcinoma. Copyright © 2014 John Wiley & Sons, Ltd.
- Published
- 2014
9. Multiple lines of evidence on the genetic relatedness of the parthenogenetic and bisexual Haemaphysalis longicornis (Acari: Ixodidae)
- Author
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Shiqi Xu, Zhijun Yu, Lida Guo, Xiaolong Yang, Xuejie Chen, Shujie Yang, Limeng Liu, and Jingze Liu
- Subjects
Microbiology (medical) ,Species complex ,Ixodidae ,Genetic Speciation ,Karyotype ,Parthenogenesis ,Population ,Genes, Insect ,Biology ,Microbiology ,Monophyly ,Phylogenetics ,Genetics ,Animals ,education ,Molecular Biology ,Phylogeny ,Ecology, Evolution, Behavior and Systematics ,education.field_of_study ,Ploidies ,Phylogenetic tree ,Chromosome ,DNA ,biology.organism_classification ,Chromosomes, Insect ,Infectious Diseases ,Evolutionary biology ,Ploidy ,Haemaphysalis longicornis - Abstract
As an obligate hematophagous ectoparasite, the hard tick Haemaphysalis longicornis exhibits two reproductive strategies, bisexual reproduction, and obligate parthenogenesis, which have attracted a widespread attention. However, the speciation of parthenogenetic population remained ambiguous due to its similarity in morphology but the remarkable differences in cytogenetics as compared with those of the bisexual ones. In the present study, we explored several new lines of genetic evidence to resolve this controversial issue. The number of the chromosomes in two lineages was checked by classical methods and their total DNA levels were determined utilizing flowcytometry. In addition, the sequences of 12S rDNA, 16S rDNA, cytochrome c oxidase I and II (COI, COII) and internal transcribed spacer-2 (ITS-2) genes were used to assess their phylogenetic relationship. We observed that the chromosome ploidy of bisexual and parthenogenetic H. longicornis collected by our laboratory was diploid and triploid, respectively. Flowcytometry analysis indicated a ratio close to 2:3 in the DNA contents of bisexual to parthenogenetic H. longicornis. Although the chromosome ploidy is different, their gene sequences are extremely similar. Analogous to the intra-species genetic difference of other invertebrates, sequence differences of all loci examined are below 2%. Phylogenetic trees constructed from 12S rDNA, 16S rDNA, COI, and ITS-2 genes revealed that they were all in the same monophyletic clade instead of splitting independently into evolutional branches. Moreover, according to 4× Rule, the K/θ ratio of two reproductive populations calculated based on COI was much smaller than four, strongly supporting that they belong to the same species. Therefore, we conclude that the evolutionary process just disturbs the chromosome ploidy and the sexual determination of parthenogenetic population and that it would be better to consider parthenogenetic H. longicornis as a metapopulation rather than a cryptic species.
- Published
- 2014
10. PS-073-Preclinical profile of HBV core protein inhibitor, ABI-H, a potent inhibitor of cccDNA generation in HBV infected cells
- Author
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Mark Bures, Haydar Simon, Walker Michael, Lida Guo, Richard J. Colonno, Xuman Tang, Ran Yan, Li Leping, Qi Huang, Xiang Xu, Yi Zhou, Dawei Cai, Rai Roopa, and Esteban Carabajal
- Subjects
Hepatology ,business.industry ,Medicine ,Core protein ,cccDNA ,business ,Virology - Published
- 2019
11. Curcumin Inhibits Proliferation and Induces Apoptosis of Human Colorectal Cancer Cells by Activating the Mitochondria Apoptotic Pathway
- Author
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Jingze Liu, Xuejie Chen, Duo Wang, Zhijun Yu, Yuhong Hu, and Lida Guo
- Subjects
Pharmacology ,chemistry.chemical_compound ,chemistry ,Annexin ,Cell culture ,Apoptosis ,Survivin ,Curcumin ,Viability assay ,Cell cycle ,Mitochondrion ,Molecular biology - Abstract
Curcumin, a natural plant extract from Curcuma longa, is known for its anti-carcinogenic and chemopreventive effects on a variety of experimental cancer models. In this study, we evaluated the effects of curcumin and elucidated its mechanism in human colorectal carcinoma cells. Cell viability assay showed that curcumin significantly inhibited the growth of LoVo cells. Curcumin treatment induced the apoptosis accompanied by ultra-structural changes and release of lactate dehydrogenase in a dose-dependent manner. Moreover, treatment with 0–30 µg/mL curcumin decreased the mitochondrial membrane potential and activated the caspase-3 and caspase-9 in a dose- and time-dependent manner. Nuclear and annexin V/PI staining showed that curcumin induced the apoptosis of LoVo cells. FACS analysis revealed that curcumin could induce the cell cycle arrest of LoVo cells at the S phase. Furthermore, western blotting analysis indicated that curcumin induced the release of cytochrome c, a significant increase of Bax and p53 and a marked reduction of Bcl-2 and survivin in LoVo cells. Taken together, our results suggested that curcumin inhibited the growth of LoVo cells by inducing apoptosis through a mitochondria-mediated pathway. Copyright © 2012 John Wiley & Sons, Ltd.
- Published
- 2012
12. Effect of bone sialoprotein on proliferation and osteodifferentiation of human bone marrow-derived mesenchymal stem cells in vitro
- Author
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Lida Guo, Zujun Jiang, Bing Xia, and Jie Wang
- Subjects
Bone sialoprotein ,medicine.medical_specialty ,Sialoglycoproteins ,Bone Marrow Cells ,Bioengineering ,Applied Microbiology and Biotechnology ,fluids and secretions ,stomatognathic system ,Internal medicine ,medicine ,Humans ,Dexamethasone ,Cell Proliferation ,DNA Primers ,Pharmacology ,Base Sequence ,General Immunology and Microbiology ,biology ,Cell growth ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,General Medicine ,Ascorbic acid ,In vitro ,Endocrinology ,biology.protein ,Osteocalcin ,Alkaline phosphatase ,Biotechnology ,medicine.drug - Abstract
We performed this study to investigate the effects of recombinant human bone sialoprotein (BSP) on the proliferation and osteodifferentiation of human BMSCs(hBMSCs). The hBMSC cultures were divided into 4 groups: control group, 10(-10) M BSP group (BSP group), osteogenic medium group (10 nM dexamethasone, 10 mM β-glycerophosphate, and 50 mg/L ascorbic acid, OM group) and BSP + OM group (OM plus10(-10) M BSP). Compared with the control group, cell growth of the other three groups slowed down, while fluorescence at the G(0)/G(1) phase increased. After 28 days, in the OM group and the BSP + OM group, the proportion of STRO-1-positive cells decreased by 22.7% and 38.4% and ALP activity increased by 50% and 71.43%, respectively. CD271 mRNA expression decreased while Cbfa1, osteocalcin and osterix mRNA levels increased in the OM and BSP + OM groups, and the mRNA level change was greater in the BSP + OM group. After 28 days, the number of nodules in the BSP + OM group was 112.5% more than that in the OM group, but nodules did not formed in the control or BSP group. We conclude that BSP is capable of inhibiting hBMSCs proliferation and enhancing their osteogenic differentiation and mineralization in the presence of OM.
- Published
- 2011
13. Osteopetrosis with micro-lacunar resorption because of defective integrin organization
- Author
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Markus Y. Mapara, Lida Guo, Alessandra Pangrazio, Lisa J. Robinson, Paul J. Orchard, Jakub Tolar, Beatrice B. Yaroslavskiy, Paolo Vezzoni, Harry C. Blair, and Ka Chen
- Subjects
Adult ,Integrins ,Podosome ,Acid Phosphatase ,Blotting, Western ,Integrin ,Osteoclasts ,Article ,Bone resorption ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Osteopetrosis, Bone resorption ,Bone Resorption ,Proto-Oncogene Proteins c-vav ,Molecular Biology ,Cells, Cultured ,030304 developmental biology ,Tartrate-resistant acid phosphatase ,Receptor activator of NF-κB ,Focal Adhesions ,0303 health sciences ,Cell fusion ,Integrin assembly ,biology ,Tartrate-Resistant Acid Phosphatase ,Cell adhesion molecule ,Cell Differentiation ,Osteopetrosis ,Blood Proteins ,Sequence Analysis, DNA ,Cell Biology ,Phosphoproteins ,medicine.disease ,Cell biology ,Isoenzymes ,src-Family Kinases ,Biochemistry ,rho GTPase ,RANKL ,030220 oncology & carcinogenesis ,Leukocytes, Mononuclear ,biology.protein ,Cell Adhesion Molecules - Abstract
In vitro differentiated monocytes were used to characterize the cellular defect in a type of osteopetrosis with minimally functional osteoclasts, in which defects associated with common causes of osteopetrosis were excluded by gene sequencing. Monocytes from the blood of a 28-year-old patient were differentiated in media with RANKL and CSF-1. Cell fusion, acid compartments within cells, and tartrate resistant acid phosphatase (TRAP) activity were normal. However, the osteoclasts made abnormally small pits on the dentine. Phalloidin labeling showed that the cell attachments lacked the peripheral ring structure that supports lacunar resorption. Instead, the osteoclasts had clusters of podosomes near the center of cell attachments. Antibody to the alphavbeta3 integrin pair or to the C-terminal of beta3 did not label podosomes, but antibody to alphav labeled them. Western blots using antibody to the N-terminal of beta3 showed a protein of reduced size. Integrins beta1 and beta5 were upregulated, but, in contrast to observations in beta3 defects, alpha2 had not increased. The rho-GTP exchange protein Vav3, a key attachment organizing protein, did not localize normally with peripheral attachment structures. Vav3 forms of 70 kD and 90 kD were identified on western blots. However, the proteins beta3 integrin, Vav3, Plekhm1, and Src, implicated in attachment defects, had normal exon sequences. In this new type of osteopetrosis, the integrin-organizing complex is dysfunctional, and at least two attachment proteins may be partially degraded.
- Published
- 2009
14. Lack of Fas antagonism by Met in human fatty liver disease
- Author
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Chunbin Zou, George K. Michalopoulos, John Stoops, Zhenqi Zhu, Carla Johnson, Lida Guo, Jihong Ma, Marie C. DeFrances, Amanda E. Eaker, Reza Zarnegar, Stephen C. Strom, and Xue Wang
- Subjects
Carcinoma, Hepatocellular ,Cirrhosis ,Molecular Sequence Data ,Apoptosis ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Fas ligand ,Jurkat Cells ,Liver disease ,Cell Line, Tumor ,Proto-Oncogene Proteins ,medicine ,Extracellular ,Humans ,Receptors, Growth Factor ,Amino Acid Sequence ,fas Receptor ,Liver Neoplasms ,Fatty liver ,General Medicine ,Proto-Oncogene Proteins c-met ,medicine.disease ,Fas receptor ,Immunohistochemistry ,Peptide Fragments ,Fatty Liver ,Kinetics ,Protein Subunits ,Biochemistry ,Hepatocyte Growth Factor Receptor ,Hepatocytes ,Cancer research ,Collagen - Abstract
Hepatocytes in fatty livers are hypersensitive to apoptosis and undergo escalated apoptotic activity via death receptor-mediated pathways, particularly that of Fas-FasL, causing hepatic injury that can eventually proceed to cirrhosis and end-stage liver disease. Here we report that the hepatocyte growth factor receptor, Met, plays an important part in preventing Fas-mediated apoptosis of hepatocytes by sequestering Fas. We also show that Fas antagonism by Met is abrogated in human fatty liver disease (FLD). Through structure-function studies, we found that a YLGA amino-acid motif located near the extracellular N terminus of the Met alpha-subunit is necessary and sufficient to specifically bind the extracellular portion of Fas and to act as a potent FasL antagonist and inhibitor of Fas trimerization. Using mouse models of FLD, we show that synthetic YLGA peptide tempers hepatocyte apoptosis and liver damage and therefore has therapeutic potential.
- Published
- 2007
15. Loss of integrin linked kinase from mouse hepatocytesin vitro andin vivo results in apoptosis and hepatitis
- Author
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René St.-Arnaud, Wendy M. Mars, Shoukat Dedhar, William C. Bowen, Yu Yang, Vasiliki Gkretsi, Chuanyue Wu, George K. Michalopoulos, Lindsay Barua, and Lida Guo
- Subjects
Programmed cell death ,Pathology ,medicine.medical_specialty ,Cellular differentiation ,Integrin ,Gene Expression ,Apoptosis ,Protein Serine-Threonine Kinases ,Biology ,Transfection ,Adenoviridae ,Hepatitis ,Hydropic degeneration ,Extracellular matrix ,Mice ,medicine ,Animals ,Integrin-linked kinase ,Phosphorylation ,Cells, Cultured ,Adaptor Proteins, Signal Transducing ,Mice, Knockout ,Integrases ,Hepatology ,Kinase ,Integrin beta1 ,Microfilament Proteins ,Membrane Proteins ,LIM Domain Proteins ,medicine.disease ,Extracellular Matrix ,Cell biology ,DNA-Binding Proteins ,Liver ,embryonic structures ,Hepatocytes ,biology.protein ,Proto-Oncogene Proteins c-akt - Abstract
Extracellular matrix (ECM) is fundamental for the survival of cells within a tissue. Loss of contact with the surrounding ECM often causes altered cell differentiation or cell death. Hepatocytes cultured without matrix lose patterns of hepatocyte-specific gene expression and characteristic cellular micro-architecture. However, differentiation is restored after the addition of hydrated matrix preparations to dedifferentiated hepatocytes. Integrin-linked kinase (ILK) is an important component of cell–ECM adhesions transmitting integrin signaling to the interior of the cell. ILK has been implicated in many fundamental cellular processes such as differentiation, proliferation, and survival. In this study, we investigated the role of ILK in mouse hepatocytes in vitro as well as in vivo. Depletion of ILK from primary mouse hepatocytes resulted in enhanced apoptosis. This was accompanied by increased caspase 3 activity and a significant decrease in expression of PINCH and α-parvin, which, along with ILK, form a stable well-characterized ternary complex at cell–ECM adhesions. The induction of apoptosis caused by ILK depletion could be substantially reversed by simultaneous overexpression of ILK, indicating that apoptosis is indeed a consequence of ILK removal. These results were further corroborated via in vivo data showing that adenoviral delivery of Cre-recombinase in ILK-floxed animals by tail vein injection resulted in acute hepatitis, with a variety of pathological findings including inflammation, fatty change, and apoptosis, abnormal mitoses, hydropic degeneration, and necrosis. Conclusion: Our results demonstrate the importance of ILK and integrin signaling for the survival of hepatocytes and the maintenance of normal liver function. (HEPATOLOGY 2007.)
- Published
- 2007
16. Formation and Phosphorylation of the PINCH-1–Integrin Linked Kinase–α-Parvin Complex Are Important for Regulation of Renal Glomerular Podocyte Adhesion, Architecture, and Survival
- Author
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Matthias Kretzler, Peter Mundel, Lida Guo, Chuanyue Wu, Simone Monika Blattner, and Yanqiang Yang
- Subjects
medicine.medical_specialty ,Cell Survival ,Renal glomerulus ,Kidney Glomerulus ,Apoptosis ,Protein Serine-Threonine Kinases ,Podocyte ,Focal adhesion ,Mice ,Internal medicine ,Cell Adhesion ,medicine ,Animals ,Ankyrin ,Actinin ,Integrin-linked kinase ,Phosphorylation ,Cells, Cultured ,Adaptor Proteins, Signal Transducing ,chemistry.chemical_classification ,biology ,Kinase ,Microfilament Proteins ,Membrane Proteins ,Cell Differentiation ,Epithelial Cells ,General Medicine ,LIM Domain Proteins ,Extracellular Matrix ,Cell biology ,DNA-Binding Proteins ,body regions ,medicine.anatomical_structure ,Endocrinology ,chemistry ,Nephrology ,embryonic structures ,biology.protein ,Ankyrin repeat - Abstract
Alterations in the cellular architecture, adhesion, and/or loss of glomerular podocytes are causal factors in the development of proteinuria and the progression to end-stage renal failure. With the use of an inducible podocyte differentiation system, it was found that the cellular levels of PINCH-1, integrin linked kinase (ILK), and alpha-parvin, cytoplasmic components of cell-extracellular matrix adhesions, were significantly increased during podocyte differentiation. Concomitantly, an increased amount of the PINCH-1-ILK-alpha-parvin complex was detected in the differentiated, foot process-containing podocytes. Overexpression of the PINCH-1-binding ankyrin repeat domain of ILK but not that of a PINCH-1-binding defective mutant form of the ankyrin domain effectively inhibited the formation of the PINCH-1-ILK-alpha-parvin complex. Disruption of the PINCH-1-ILK-alpha-parvin complex significantly reduced the podocyte-matrix adhesion and foot process formation. Furthermore, a marked increase of apoptosis in the podocytes in which the assembly of the PINCH-1-ILK-alpha-parvin complex was compromised was detected. Inhibition of ILK with a small compound inhibitor also altered podocyte cytoskeleton and increased apoptosis. Finally, it is shown that alpha-parvin is phosphorylated in podocytes. Mutations at the alpha-parvin N-terminal proline-directed serine phosphorylation sites reduced its complex formation with ILK and resulted in defects in podocyte adhesion, architecture, and survival. These results provide important evidence for a crucial role of the PINCH-1-ILK-alpha-parvin complex in the control of podocyte adhesion, morphology, and survival.
- Published
- 2005
17. CH-ILKBP regulates cell survival by facilitating the membrane translocation of protein kinase B/Akt
- Author
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Tomohiko Fukuda, Xiaohua Shi, Chuanyue Wu, and Lida Guo
- Subjects
Cell Survival ,Green Fluorescent Proteins ,AKT1 ,Apoptosis ,Protein Serine-Threonine Kinases ,Biology ,DEPTOR ,mTORC2 ,03 medical and health sciences ,0302 clinical medicine ,Proto-Oncogene Proteins ,Report ,apoptosis ,CH-ILKBP ,protein kinase B/Akt ,membrane translocation ,ILK ,Tumor Cells, Cultured ,Humans ,Actinin ,Phosphorylation ,RNA, Small Interfering ,Protein kinase B ,PI3K/AKT/mTOR pathway ,030304 developmental biology ,Mitogen-Activated Protein Kinase 1 ,0303 health sciences ,Mitogen-Activated Protein Kinase 3 ,Akt/PKB signaling pathway ,Cell Membrane ,Microfilament Proteins ,Biological Transport ,Cell Biology ,Extracellular Matrix ,Cell biology ,Enzyme Activation ,Luminescent Proteins ,Caspases ,030220 oncology & carcinogenesis ,Mitogen-Activated Protein Kinases ,Signal transduction ,Carrier Proteins ,Proto-Oncogene Proteins c-akt ,HeLa Cells ,Protein Binding - Abstract
Cell survival depends on proper propagation of protective signals through intracellular signaling intermediates. We report here that calponin homology domain–containing integrin-linked kinase (ILK)–binding protein (CH-ILKBP), a widely expressed adaptor protein localized at plasma membrane-actin junctions, is essential for transmission of survival signals. Cells that are depleted of CH-ILKBP undergo extensive apoptosis despite the presence of cell–extracellular matrix contacts and soluble growth factors. The activating phosphorylation of protein kinase B (PKB/Akt), a key regulator of apoptosis, is impaired in the absence of CH-ILKBP. Importantly, loss of CH-ILKBP prevents the membrane translocation of PKB/Akt. Furthermore, forced membrane targeting of PKB/Akt bypasses the requirement of CH-ILKBP for the activating phosphorylation of PKB/Akt, suggesting that CH-ILKBP is required for the membrane translocation but not the subsequent phosphorylation of PKB/Akt. Finally, we show that loss of CH-ILKBP is also required for the full activation of extracellular signal–regulated kinase (ERK)1/2. However, restoration of the PKB/Akt activation is sufficient for protection of cells from apoptosis induced by the depletion of CH-ILKBP despite the persistent suppression of the ERK1/2 activation. Thus, CH-ILKBP is an important component of the prosurvival signaling pathway functioning primarily by facilitating the membrane translocation of PKB/Akt and consequently the activation of PKB/Akt in response to extracellular survival signals.
- Published
- 2003
18. A Critical Role of the PINCH-Integrin-linked Kinase Interaction in the Regulation of Cell Shape Change and Migration
- Author
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Yongjun Zhang, Chuanyue Wu, Lida Guo, and Ka Chen
- Subjects
chemistry.chemical_classification ,Binding Sites ,biology ,Signal transducing adaptor protein ,Motility ,Cell Biology ,Protein Serine-Threonine Kinases ,Actin cytoskeleton ,Biochemistry ,Cell biology ,DNA-Binding Proteins ,body regions ,Extracellular matrix ,Focal adhesion ,chemistry ,Cell Movement ,embryonic structures ,Cell Adhesion ,biology.protein ,Ankyrin ,Integrin-linked kinase ,Molecular Biology ,Cell Size ,LIM domain - Abstract
The interaction of cells with extracellular matrix recruits multiple proteins to cell-matrix contact sites (e.g. focal and fibrillar adhesions), which connect the extracellular matrix to the actin cytoskeleton and regulate cell shape change, migration, and other cellular processes. We previously identified PINCH, an adaptor protein comprising primarily five LIM domains, as a binding protein for integrin-linked kinase (ILK). In this study, we show that PINCH co-localizes with ILK in both focal adhesions and fibrillar adhesions. Furthermore, we have investigated the molecular basis underlying the targeting of PINCH to the cell-matrix contact sites and the functional significance of the PINCH-ILK interaction. We have found that the N-terminal LIM1 domain, which mediates the ILK binding, is required for the targeting of PINCH to the cell-matrix contact sites. The C-terminal LIM domains, although not absolutely required, play an important regulatory role in the localization of PINCH to cell-matrix contact sites. Inhibition of the PINCH-ILK interaction, either by overexpression of a PINCH N-terminal fragment containing the ILK-binding LIM1 domain or by overexpression of an ILK N-terminal fragment containing the PINCH-binding ankyrin domain, retarded cell spreading, and reduced cell motility. These results suggest that PINCH, through its interaction with ILK, is crucially involved in the regulation of cell shape change and motility.
- Published
- 2002
19. A novel Death Defying Domain in Met entraps the active site of Caspase-3 and blocks apoptosis in hepatocytes
- Author
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Xinping Tan, Reza Zarnegar, Danushka S. Seneviratne, Jihong Ma, Marie C. DeFrances, Jiyan An, Chunbin Zou, Lida Guo, Robert Bowser, and Yong-Kook Kwon
- Subjects
Molecular Sequence Data ,Caspase 3 ,Apoptosis ,Cleavage (embryo) ,Receptor tyrosine kinase ,Article ,Mice ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Caspase ,Binding Sites ,Hepatology ,biology ,Proto-Oncogene Proteins c-met ,equipment and supplies ,Caspase Inhibitors ,Cell biology ,Protein Structure, Tertiary ,Biochemistry ,Cell culture ,Cytoprotection ,biology.protein ,Hepatocytes ,Hepatocyte growth factor ,Oligopeptides ,medicine.drug - Abstract
Met, the transmembrane tyrosine kinase receptor for hepatocyte growth factor (HGF), is known to function as a potent antiapoptotic mediator in normal and neoplastic cells. Herein we report that the intracellular cytoplasmic tail of Met has evolved to harbor a tandem pair of caspase-3 cleavage sites, which bait, trap, and disable the active site of caspase-3, thereby blocking the execution of apoptosis. We call this caspase-3 cleavage motif the Death Defying Domain (DDD). This site consists of the following sequence: DNAD-DEVD-T (where the hyphens denote caspase cleavage sites). Through functional and mechanistic studies, we show that upon DDD cleavage by caspase-3 the resulting DEVD-T peptide acts as a competitive inhibitor and entraps the active site of caspase-3 akin to DEVD-CHO, which is a potent, synthetic inhibitor of caspase-3 activity. By gain- and loss-of-function studies using restoration of DDD expression in DDD-deficient hepatocytic cells, we found that both caspase-3 sites in DDD are necessary for inhibition of caspase-3 and promotion of cell survival. Employing mutagenesis studies, we show that DDD could operate independently of Met's enzymatic activity as determined by using kinase-dead human Met mutant constructs. Studies of both human liver cancer tissues and cell lines uncovered that DDD cleavage and entrapment of caspase-3 by DDD occur in vivo, further proving that this site has physiological and pathophysiological relevance. Conclusion: Met can directly inhibit caspase-3 by way of a novel mechanism and promote hepatocyte survival. The results presented here will further our understanding of the mechanisms that control not only normal tissue homeostasis but also abnormal tissue growth such as cancer and degenerative diseases in which apoptotic caspases are at play. (Hepatology 2014;59:2010–2021)
- Published
- 2014
20. Elevated expression of axin2 and hnkd mRNA provides evidence that Wnt/β-catenin signaling is activated in human colon tumors
- Author
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Ann B. Jefferson, Jaime Escobedo, Christoph Reinhard, Doreen Sakamoto, Dong Yan, Roger H. Caothien, Lewis T. Williams, Lida Guo, Filippo Randazzo, Vivien W. Chan, Marion Wiesmann, Michael Rohan, John Fuller, Wendy J. Fantl, and Pablo Garcia
- Subjects
DNA, Complementary ,Beta-catenin ,Adenomatous polyposis coli ,Molecular Sequence Data ,Biology ,Cell Line ,Axin Protein ,Proto-Oncogene Proteins ,medicine ,AXIN2 ,Humans ,RNA, Messenger ,Promoter Regions, Genetic ,beta Catenin ,DNA Primers ,Oligonucleotide Array Sequence Analysis ,Multidisciplinary ,Base Sequence ,Wnt signaling pathway ,Proteins ,Cancer ,Transfection ,Zebrafish Proteins ,Biological Sciences ,medicine.disease ,Molecular biology ,Repressor Proteins ,Wnt Proteins ,Naked cuticle ,Cytoskeletal Proteins ,Cell culture ,Colonic Neoplasms ,Trans-Activators ,biology.protein ,Signal Transduction - Abstract
Genetic studies have identified mutations in key regulators of the Wnt/β-catenin pathway in a variety of cancers, most frequently in colon cancers. However, whether the pathway is activated in clinical cancer samples is not easily determined, and therefore it is useful to find markers that could be surrogates to show activation of the Wnt/β-catenin pathway. Gene expression profiles were analyzed in SW620, a colon cancer cell line in which β-catenin levels are stabilized as a consequence of truncated adenomatous polyposis coli and were compared with profiles of the same cells transfected with antisense oligodeoxynucleotides. Treatment of cells with β-catenin antisense oligodeoxynucleotides resulted in a decrease in the levels of axin2 and human naked cuticle ( hnkd ) mRNAs. Interestingly, the proteins encoded by both of these mRNAs are known inhibitors of the β-catenin pathway. In 30 human cell lines derived from different origins, axin2 and hnkd were expressed only in human colon cancer cell lines that are known to have activating mutations in the Wnt/β-catenin pathway. Further, levels of both axin2 and hnkd mRNA were also found to be elevated in about 65% of laser microdissected cells from human colon tumors compared with laser microdissected cells of normal morphology from the same patient samples. The increased expression of axin2 and hnkd correlated with truncations in adenomatous polyposis coli in the same patient samples. These results reveal that it is possible to detect activation of a carcinogenic pathway in human cancer samples with specific markers.
- Published
- 2001
21. The Distribution and Regulation of Integrin-Linked Kinase in Normal and Diabetic Kidneys
- Author
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Paul W. Sanders, Chuanyue Wu, Lida Guo, and Anne Woods
- Subjects
medicine.medical_specialty ,Kidney Glomerulus ,Integrin ,Protein Serine-Threonine Kinases ,Kidney ,Pathology and Forensic Medicine ,Extracellular matrix ,Reference Values ,Internal medicine ,medicine ,Animals ,Humans ,Diabetic Nephropathies ,Tissue Distribution ,Integrin-linked kinase ,Cell adhesion ,biology ,Glomerulosclerosis, Focal Segmental ,Cell adhesion molecule ,Glomerulosclerosis ,medicine.disease ,Extracellular Matrix ,Fibronectins ,Glomerular Mesangium ,Rats ,Fibronectin ,Endocrinology ,Mesangium ,embryonic structures ,biology.protein ,Regular Articles - Abstract
Alteration in cell adhesion and extracellular matrix deposition is a hallmark of diabetic glomerulosclerosis. Integrin-linked kinase (ILK) is a recently identified integrin cytoplasmic-binding protein that has been implicated in the regulation of cell adhesion and extracellular matrix deposition. To begin to investigate whether ILK is involved in the pathogenesis of diabetic glomerulosclerosis, we have analyzed the distribution and regulation of ILK in normal and diabetic kidneys as well as in isolated mesangial cells. We have found that ILK is normally expressed at high concentration in visceral epithelial cells. In diabetic glomeruli, ILK expression in the mesangium is dramatically increased. The increase in ILK level is associated with diffuse mesangial expansion. In glomeruli where advanced nodular sclerosis and global sclerosis were dominant, ILK level was reduced, suggesting that the increase in ILK expression likely associates with relatively early glomerulosclerosis. Additionally, we have found that exposure of mesangial cells to high concentrations of glucose significantly increased the ILK level. Finally, we show that ILK localizes to regions of cell membranes that are in close contact with mesangial fibronectin matrix. These results suggest that ILK is likely involved in mesangial matrix expansion in response to hyperglycemia in the pathogenesis of diabetic glomerulosclerosis.
- Published
- 2001
22. Development of anti-p185HER2 immunoliposomes for cancer therapy
- Author
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C Wirth, H Asgari, Keelung Hong, Paul Carter, W I Wood, G A Keller, C Kotts, R Shalaby, Lida Guo, and John W. Park
- Subjects
Receptor, ErbB-2 ,medicine.drug_class ,media_common.quotation_subject ,Mice, SCID ,In Vitro Techniques ,Monoclonal antibody ,Mice ,In vivo ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Doxorubicin ,Internalization ,media_common ,Multidisciplinary ,biology ,Antibodies, Monoclonal ,Cancer ,medicine.disease ,Molecular biology ,Endocytosis ,Growth Inhibitors ,In vitro ,Liposomes ,biology.protein ,Female ,Antibody ,Research Article ,medicine.drug - Abstract
The product of the HER2 protooncogene, p185HER2, represents an attractive target for cancer immunotherapies. We have prepared anti-p185HER2 immunoliposomes in which Fab' fragments of a humanized anti-p185HER2 monoclonal antibody with antiproliferative properties (rhuMAb-HER2) were conjugated to either conventional or sterically stabilized liposomes. These immunoliposomes bind specifically to p185HER2-overexpressing breast cancer cells (SK-BR-3 and BT-474). High-affinity binding of anti-p185HER2 immunoliposomes is comparable to that of free rhuMAbHER2-Fab' or the intact antibody. Empty immunoliposomes inhibit the culture growth of p185HER2-overexpressing breast cancer cells, and this antiproliferative effect is superior to that of free rhuMAbHER2-Fab', indicating that liposomal anchoring of these anti-p185HER2 Fab' fragments enhances their biological activity. Efficient internalization of anti-p185HER2 immunoliposomes, demonstrated by light and electron microscopy, occurs by receptor-mediated endocytosis via the coated pit pathway and also possibly by membrane fusion. Doxorubicin-loaded anti-p185HER2 immunoliposomes are markedly and specifically cytotoxic against p185HER2-overexpressing tumor cells in vitro. Anti-p185HER2 immunoliposomes administered in vivo in Scid mice bearing human breast tumor (BT-474) xenografts can deliver doxorubicin to tumors. These results indicate that anti-p185HER2 immunoliposomes are a promising therapeutic vehicle for the treatment of p185HER2-overexpressing human cancers.
- Published
- 1995
23. Preclinical Characterization of Potent Core Protein Assembly Modulators for the Treatment of Chronic Hepatitis B
- Author
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T. Sun, G. Renuka Kumar, S. Katen, J. Deer, Jing Liu, P.-C. Li, Lida Guo, Sheena Francis, E. Connelly, Kirk R. Henne, Qixing Huang, S. Sun, W.W. Turner, Cathal Mahon, G. Chen, L.D. Arnold, E. May, Yuhua Zong, Adam Zlotnick, Yi Zhou, Mark Bures, U. Lopatin, Li Leping, K. Nabel, and A. Mercier
- Subjects
03 medical and health sciences ,0302 clinical medicine ,Hepatology ,Chronic hepatitis ,business.industry ,Medicine ,030211 gastroenterology & hepatology ,Core protein ,030204 cardiovascular system & hematology ,Pharmacology ,business ,Virology - Published
- 2016
24. Na+/H+ exchanger regulatory factor 1 (NHERF1) directly regulates osteogenesis
- Author
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Lida Guo, Harry C. Blair, Irina L. Tourkova, Li Liu, Sarah E. Henderson, Verónica Alonso, Peter A. Friedman, and Alejandro J. Almarza
- Subjects
medicine.medical_specialty ,Sodium-Hydrogen Exchangers ,Cellular differentiation ,Bone Matrix ,Osteoclasts ,Biochemistry ,Bone resorption ,Extracellular matrix ,chemistry.chemical_compound ,Mice ,Calcification, Physiologic ,Osteoclast ,Osteogenesis ,Internal medicine ,Adipocyte ,medicine ,Animals ,Humans ,Molecular Biology ,Osteomalacia ,Osteoblasts ,Chemistry ,Osteoblast ,Cell Differentiation ,Mesenchymal Stem Cells ,Molecular Bases of Disease ,Cell Biology ,medicine.disease ,Phosphoproteins ,Mice, Mutant Strains ,Extracellular Matrix ,Sodium–hydrogen antiporter ,medicine.anatomical_structure ,Endocrinology - Abstract
Bone formation requires synthesis, secretion, and mineralization of matrix. Deficiencies in these processes produce bone defects. The absence of the PDZ domain protein Na(+)/H(+) exchange regulatory factor 1 (NHERF1) in mice, or its mutation in humans, causes osteomalacia believed to reflect renal phosphate wasting. We show that NHERF1 is expressed by mineralizing osteoblasts and organizes Na(+)/H(+) exchangers (NHEs) and the PTH receptor. NHERF1-null mice display reduced bone formation and wide mineralizing fronts despite elimination of phosphate wasting by dietary supplementation. Bone mass was normal, reflecting coordinated reduction of bone resorption and formation. NHERF1-null bone had decreased strength, consistent with compromised matrix quality. Mesenchymal stem cells from NHERF1-null mice showed limited osteoblast differentiation but enhanced adipocyte differentiation. PTH signaling and Na(+)/H(+) exchange were dysregulated in these cells. Osteoclast differentiation from monocytes was unaffected. Thus, NHERF1 is required for normal osteoblast differentiation and matrix synthesis. In its absence, compensatory mechanisms maintain bone mass, but bone strength is reduced.
- Published
- 2012
25. Blocking antibody to the β-subunit of FSH prevents bone loss by inhibiting bone resorption and stimulating bone synthesis
- Author
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Irina L. Tourkova, Alberta Zallone, Clifford J. Rosen, Jay Cao, Lida Guo, Tony Yuen, Li Sun, Rauf Latif, Harry C. Blair, Maria I. New, Terry F. Davies, Zhuan Bian, Ling-Ling Zhu, Mone Zaidi, and Jianhua Li
- Subjects
medicine.medical_specialty ,endocrine system ,Ovariectomy ,Osteoporosis ,Osteoclasts ,Biology ,Bone resorption ,Antibodies ,Colony-Forming Units Assay ,Follicle-stimulating hormone ,Mice ,Osteoclast ,Internal medicine ,Blocking antibody ,medicine ,Animals ,Humans ,Osteoporosis, Postmenopausal ,Mice, Knockout ,Multidisciplinary ,Bone Development ,Osteoblast ,Mesenchymal Stem Cells ,Biological Sciences ,medicine.disease ,Bone morphogenetic protein 7 ,medicine.anatomical_structure ,Endocrinology ,Follicle Stimulating Hormone, beta Subunit ,Receptors, FSH ,Female ,Follicle-stimulating hormone receptor ,hormones, hormone substitutes, and hormone antagonists - Abstract
Low estrogen levels undoubtedly underlie menopausal bone thinning. However, rapid and profuse bone loss begins 3 y before the last menstrual period, when serum estrogen is relatively normal. We have shown that the pituitary hormone FSH, the levels of which are high during late perimenopause, directly stimulates bone resorption by osteoclasts. Here, we generated and characterized a polyclonal antibody to a 13-amino-acid-long peptide sequence within the receptor-binding domain of the FSH β-subunit. We show that the FSH antibody binds FSH specifically and blocks its action on osteoclast formation in vitro. When injected into ovariectomized mice, the FSH antibody attenuates bone loss significantly not only by inhibiting bone resorption, but also by stimulating bone formation, a yet uncharacterized action of FSH that we report herein. Mesenchymal cells isolated from mice treated with the FSH antibody show greater osteoblast precursor colony counts, similarly to mesenchymal cells isolated from FSH receptor ( FSHR ) −/− mice. This suggests that FSH negatively regulates osteoblast number. We confirm that this action is mediated by signaling-efficient FSHRs present on mesenchymal stem cells. Overall, the data prompt the future development of an FSH-blocking agent as a means of uncoupling bone formation and bone resorption to a therapeutic advantage in humans.
- Published
- 2012
26. Regulation of fibronectin matrix deposition and cell proliferation by the PINCH‐ILK‐CH‐ILKBP complex
- Author
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Lida Guo and Chuanyue Wu
- Subjects
medicine.medical_specialty ,Cell division ,Macromolecular Substances ,Glomerular Mesangial Cell ,Actinin ,Protein Serine-Threonine Kinases ,Transfection ,Models, Biological ,Biochemistry ,Internal medicine ,Cell Adhesion ,Genetics ,medicine ,Animals ,Integrin-linked kinase ,Cell adhesion ,Molecular Biology ,Cells, Cultured ,biology ,Cell growth ,Chemistry ,Microfilament Proteins ,Peptide Fragments ,Extracellular Matrix ,Fibronectins ,Glomerular Mesangium ,Rats ,Cell biology ,DNA-Binding Proteins ,body regions ,Fibronectin ,Endocrinology ,embryonic structures ,biology.protein ,Carrier Proteins ,Cell Division ,Biotechnology - Abstract
Alteration in renal glomerular mesangial cell growth and fibronectin matrix deposition is a hallmark of glomerulosclerosis, which ultimately leads to end-stage renal failure. We have previously shown that the expression of integrin-linked kinase (ILK), a cytoplasmic component of the cell-extracellular matrix contacts, is increased in mesangial cells in human patients with diabetic nephropathy. We show here that ILK forms a complex with PINCH and CH-ILKBP in primary mesangial cells, which are co-clustered at fibrillar adhesions, sites that are involved in fibronectin matrix deposition. To investigate functional significance of the PINCH-ILK-CH-ILKBP complex formation, we expressed the PINCH-binding N-terminal fragment and the CH-ILKBP-binding C-terminal fragment of ILK, respectively, in mesangial cells by using an adenoviral expression system. Overexpression of either the N-terminal fragment or the C-terminal fragment of ILK effectively inhibited the PINCH-ILK-CH-ILKBP complex formation. Inhibition of the PINCH-ILK-CH-ILKBP complex formation significantly reduced fibronectin matrix deposition and inhibited cell proliferation. These results indicate that the PINCH-ILK-CH-ILKBP complex is critically involved in the regulation of mesangial fibronectin matrix deposition and cell proliferation, and suggest that it may potentially serve as a useful target in the therapeutic control of progressive renal failure and other pathological processes involving abnormal cell proliferation and fibronectin matrix deposition.
- Published
- 2002
27. Life cycle of Haemaphysalis doenitzi (Acari: Ixodidae) under laboratory conditions and its phylogeny based on mitochondrial 16S rDNA
- Author
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Hao Meng, Jingze Liu, Xuejie Chen, Lingxia Li, Duo Wang, Zhijun Yu, Lida Guo, and Renjie Liu
- Subjects
Male ,Ixodidae ,Oviposition ,Zoology ,Biology ,DNA, Mitochondrial ,RNA, Ribosomal, 16S ,Animals ,Acari ,Nymph ,Phylogeny ,Larva ,Life Cycle Stages ,Ecology ,Reproductive success ,Base Sequence ,Hatching ,Body Weight ,General Medicine ,Feeding Behavior ,Sequence Analysis, DNA ,biology.organism_classification ,Animal ecology ,Insect Science ,Female ,Moulting - Abstract
The paper investigated the life cycle of the hard tick Haemaphysalis doenitzi under laboratory conditions and its phylogeny based on mitochondrial 16S rDNA. The results revealed that the complete life cycle of H. doenitzi requires a mean duration of 109.6 days ranging from 91 to 137 days and the average prefeeding, feeding and premoulting periods of larvae, nymphs and females and the eggs hatching period are 18.7, 26.9, 38.9, and 25.1 days, respectively. In addition, the weight of engorged females is highly correlated with the number of egg masses laid (r = 0.936, P
- Published
- 2011
28. Variation in human erythrocyte membrane unsaturated Fatty acids: correlation with cardiovascular disease
- Author
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Jorge L. Sepulveda, Yvette C. Tanhehco, Monica Frey, Lida Guo, Lorna J. Cropcho, K. Michael Gibson, and Harry C. Blair
- Subjects
Adult ,Erythrocyte Membrane ,Troponin I ,Palmitic Acid ,General Medicine ,Cholesterol, LDL ,Middle Aged ,Pathology and Forensic Medicine ,Medical Laboratory Technology ,Cardiovascular Diseases ,Risk Factors ,Case-Control Studies ,Fatty Acids, Unsaturated ,Humans ,lipids (amino acids, peptides, and proteins) ,Biomarkers ,Stearic Acids - Abstract
Context.—Whether cell membrane fatty acid (FA) composition is a useful indicator of vascular disease is unclear. Objective.—To study variation of erythrocyte (RBC) membrane FA in samples from healthy volunteers, hospitalized patients, and cardiac troponin I–elevated patients with myocardial damage without a priori assumptions as to FA composition. Design.—We separated FAs extracted from RBCs by gas chromatography and identified them by mass spectrometry. Fatty acids with abundance greater than 1% of total were quantified and compared: hexadecanoic (C16:0), octadecadienoic (C18:2), cis- and trans-octadecenoic (C18:1), and eicosatetraenoic (C20:4) acids. Deuterated standards established proportionality of FA recovery. The cis- and trans-C18:1 identification was verified by comparison with standards. Results.—In troponin-positive samples, C18:2 to C18:1 ratios were increased 30% compared with healthy controls or with random patient samples. Erythrocyte trans-C18:1 had a wide variation, ∼10-fold, in all groups but without differences between groups. Replicates showed that the wide range of RBC trans-FA load is not due to analytic variation. In healthy subjects, the RBC content of lower– molecular weight FAs (C16-C18) correlated with serum low-density lipoprotein cholesterol, but despite the established relationship between dietary trans-FA and increased low-density lipoprotein cholesterol, lipid profiles had no correlation with RBC trans-FA content. Conclusions.—Erythrocyte accumulation of unsaturated FA may be a useful indicator of vascular disease, whereas the wide range in trans-FAs suggests that both diet and genetic variation affect RBC trans-FA accumulation. Unsaturated FAs increase membrane fluidity and may reflect a natural response to subclinical vascular changes, which may in turn reflect increased risk of clinical disease.
- Published
- 2010
29. Estrogen Inhibits RANKL-stimulated Osteoclastic Differentiation of Human Monocytes through Estrogen and RANKL-regulated Interaction of Estrogen Receptor-α with BCAR1 and Traf6
- Author
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Lida Guo, Harry C. Blair, Lisa J. Robinson, Irina L. Tourkova, Reed D. Griswold, Eva V. Zadorozny, and Beatrice B. Yaroslavskiy
- Subjects
musculoskeletal diseases ,medicine.medical_specialty ,medicine.drug_class ,Cell Survival ,Cellular differentiation ,Lipopolysaccharide Receptors ,Osteoclasts ,Biology ,Article ,Monocytes ,Osteoclast ,Internal medicine ,medicine ,Animals ,Humans ,RNA, Small Interfering ,Estrogen receptor beta ,Cells, Cultured ,TNF Receptor-Associated Factor 6 ,RANK Ligand ,Estrogen Receptor alpha ,NF-kappa B ,Cell Differentiation ,Estrogens ,Cell Biology ,Endocrinology ,medicine.anatomical_structure ,Crk-Associated Substrate Protein ,Estrogen ,RANKL ,biology.protein ,Female ,Signal transduction ,Estrogen receptor alpha ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction - Abstract
The effects of estrogen on osteoclast survival and differentiation were studied using CD14-selected mononuclear osteoclast precursors from peripheral blood. Estradiol at approximately 1 nM reduced RANKL-dependent osteoclast differentiation by 40-50%. Osteoclast differentiation was suppressed 14 days after addition of RANKL even when estradiol was withdrawn after 18 h. In CD14+ cells apoptosis was rare and was not augmented by RANKL or by 17-beta-estradiol. Estrogen receptor-alpha (ERalpha) expression was strongly down-regulated by RANKL, whether or not estradiol was present. Mature human osteoclasts thus cannot respond to estrogen via ERalpha. However, ERalpha was present in CD14+ osteoclast progenitors, and a scaffolding protein, BCAR1, which binds ERalpha in the presence of estrogen, was abundant. Immunoprecipitation showed rapid (approximately 5 min) estrogen-dependent formation of ERalpha-BCAR1 complexes, which were increased by RANKL co-treatment. The RANKL-signaling intermediate Traf6, which regulates NF-kappaB activity, precipitated with this complex. Reduction of NF-kappaB nuclear localization occurred within 30 min of RANKL stimulation, and estradiol inhibited the phosphorylation of IkappaB in response to RANKL. Inhibition by estradiol was abolished by siRNA knockdown of BCAR1. We conclude that estrogen directly, but only partially, curtails human osteoclast formation. This effect requires BCAR1 and involves a non-genomic interaction with ERalpha.
- Published
- 2009
30. Role of the integrin-linked kinase/PINCH1/alpha-parvin complex in cardiac myocyte hypertrophy
- Author
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Wen Yan, Xueyin N. Huang, Shoukat Dedhar, Lekha Tummalapali, Lida Guo, René St-Arnaud, Hua Chen, Ka Chen, Chuanyue Wu, and Jorge L. Sepulveda
- Subjects
medicine.medical_specialty ,Heart Ventricles ,Integrin ,Cell Culture Techniques ,Biology ,Protein Serine-Threonine Kinases ,Pathology and Forensic Medicine ,Muscle hypertrophy ,Adenoviridae ,Rats, Sprague-Dawley ,Mice ,Phenylephrine ,Atrial natriuretic peptide ,Internal medicine ,medicine ,Myocyte ,Animals ,Integrin-linked kinase ,Actinin ,Myocytes, Cardiac ,RNA, Messenger ,Molecular Biology ,Cells, Cultured ,Adaptor Proteins, Signal Transducing ,Cell adhesion molecule ,Myocardium ,Cardiac myocyte ,Membrane Proteins ,Proteins ,Drug Synergism ,Cell Biology ,Hypertrophy ,LIM Domain Proteins ,Cell biology ,Fibronectins ,Rats ,DNA-Binding Proteins ,Endocrinology ,Animals, Newborn ,embryonic structures ,biology.protein ,Signal transduction ,Adrenergic alpha-Agonists ,Atrial Natriuretic Factor - Abstract
Outside-in signaling from fibronectin (FN) through integrin receptors has been shown to play an important role in promoting cardiac myocyte hypertrophy and synergizes with other hypertrophic stimuli such as the alpha-adrenergic agonist phenylephrine (PE) and mechanical strain. The integrin-linked kinase (ILK) is a critical molecule involved in cell adhesion, motility and survival in nonmyocytes such as fibroblasts and epithelial cells. Its role in cardiac myocytes is unclear. In this study, we demonstrate that (1) ILK forms a complex with PINCH1 and alpha-parvin proteins (IPAP1 complex) in neonatal rat ventricular myocytes; (2) localization of IPAP1 complex proteins to costameres in cardiac myocytes is stimulated by FN, PE and synergistically by the combination of FN and PE in an integrin beta1-dependent manner; (3) a dominant-negative mutant lacking the PINCH-binding N-terminus of ILK (ILK-C) prevents costamere association of ILK and alpha-parvin, but not PINCH1; (4) FN- and PE-induced hypertrophy, measured by increased protein/DNA ratio, beating frequency and atrial natriuretic peptide expression, is stimulated by low levels of ILK-C but repressed by high ILK-C expression; and (5) overexpression of ILK-C, as well as deletion of the ILK gene in mouse neonatal ventricular myocytes, induces marked apoptosis of cardiac myocytes. These results suggest that the IPAP1 complex plays an important role in mediating integrin-signaling pathways that regulate cardiac myocyte hypertrophy and resistance to apoptosis.
- Published
- 2005
31. beta-Catenin regulates vascular endothelial growth factor expression in colon cancer
- Author
-
Vijay, Easwaran, Sang H, Lee, Landon, Inge, Lida, Guo, Cheryl, Goldbeck, Evelyn, Garrett, Marion, Wiesmann, Pablo D, Garcia, John H, Fuller, Vivien, Chan, Filippo, Randazzo, Robert, Gundel, Robert S, Warren, Jaime, Escobedo, Sharon L, Aukerman, Robert N, Taylor, and Wendy J, Fantl
- Subjects
Vascular Endothelial Growth Factor A ,Genes, APC ,Colon ,Recombinant Fusion Proteins ,Endothelial Growth Factors ,Adenocarcinoma ,Transfection ,Oligodeoxyribonucleotides, Antisense ,Mice ,Tumor Cells, Cultured ,Animals ,Humans ,RNA, Messenger ,RNA, Neoplasm ,Intestinal Mucosa ,Growth Substances ,Promoter Regions, Genetic ,beta Catenin ,Lymphokines ,Binding Sites ,Vascular Endothelial Growth Factors ,Mice, Mutant Strains ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Mice, Inbred C57BL ,Cytoskeletal Proteins ,Genes, ras ,Adenomatous Polyposis Coli ,Colonic Neoplasms ,Trans-Activators ,Intercellular Signaling Peptides and Proteins ,Fibroblast Growth Factor 2 ,Signal Transduction ,Subcellular Fractions - Abstract
To evaluate whether beta-catenin signaling has a role in the regulation of angiogenesis in colon cancer, a series of angiogenesis-related gene promoters was analyzed for beta-catenin/TCF binding sites. Strikingly, the gene promoter of human vascular endothelial growth factor (VEGF, or VEGF-A) contains seven consensus binding sites for beta-catenin/TCF. Analysis of laser capture microdissected human colon cancer tissue indicated a direct correlation between up-regulation of VEGF-A expression and adenomatous polyposis coli (APC) mutational status (activation of beta-catenin signaling) in primary tumors. In metastases, this correlation was not observed. Analysis by immunohistochemistry of intestinal polyps in mice heterozygous for the multiple intestinal neoplasia gene (Min/+) at 5 months revealed an increase and redistribution of VEGF-A in proximity to those cells expressing nuclear beta-catenin with a corresponding increase in vessel density. Transfection of normal colon epithelial cells with activated beta-catenin up-regulated levels of VEGF-A mRNA and protein by 250-300%. When colon cancer cells with elevated beta-catenin levels were treated with beta-catenin antisense oligodeoxynucleotides, VEGF-A expression was reduced by more than 50%. Taken together, our observations indicate a close link between beta-catenin signaling and the regulation of VEGF-A expression in colon cancer.
- Published
- 2003
32. Characterization of PINCH-2, a new focal adhesion protein that regulates the PINCH-1-ILK interaction, cell spreading, and migration
- Author
-
Lida Guo, Yongjun Zhang, Ka Chen, and Chuanyue Wu
- Subjects
Recombinant Fusion Proteins ,Mutant ,Molecular Sequence Data ,Matrix (biology) ,Biology ,Protein Serine-Threonine Kinases ,Biochemistry ,Cell Line ,Focal adhesion ,Mice ,Cell Movement ,Genes, Reporter ,medicine ,Cell Adhesion ,Animals ,Humans ,Molecular Biology ,Adaptor Proteins, Signal Transducing ,Cell Size ,Focal Adhesions ,Kinase ,Wild type ,Membrane Proteins ,Nuclear Proteins ,Cell Biology ,Adhesion ,LIM Domain Proteins ,Immunohistochemistry ,Cell biology ,Extracellular Matrix ,Protein Structure, Tertiary ,body regions ,DNA-Binding Proteins ,medicine.anatomical_structure ,embryonic structures ,biology.protein ,Antibody ,Carrier Proteins ,Nucleus - Abstract
Integrin-linked kinase (ILK) is a multidomain protein that plays important roles at cell-extracellular matrix (ECM) adhesion sites. We describe here a new LIM-domain containing protein (termed as PINCH-2) that forms a complex with ILK. PINCH-2 is co-expressed with PINCH-1 (previously known as PINCH), another member of the PINCH protein family, in a variety of human cells. Immunofluorescent staining of cells with PINCH-2-specific antibodies show that PINCH-2 localizes to both cell-ECM contact sites and the nucleus. Deletion of the first LIM (LIM1) domain of PINCH-2 abolished the ability of PINCH-2 to form a complex with ILK. The ILK-binding defective LIM1-deletion mutant, unlike the wild type PINCH-2 or the ILK-binding competent LIM5-deletion mutant, was incapable of localizing to cell-ECM contact sites, suggesting that ILK binding is required for this process. Importantly, the PINCH-2-ILK and PINCH-1-ILK interactions are mutually exclusive. Overexpression of PINCH-2 significantly inhibited the PINCH-1-ILK interaction and reduced cell spreading and migration. These results identify a novel nuclear and focal adhesion protein that associates with ILK and reveals an important role of PINCH-2 in the regulation of the PINCH-1-ILK interaction, cell shape change, and migration.
- Published
- 2002
33. Vitamin C Prevents Hypogonadal Bone Loss
- Author
-
Harry C. Blair, Zhuan Bian, Liu Peng, Jeffrey I. Mechanick, Mone Zaidi, Jianhua Li, Ling-Ling Zhu, Merry Sun, Jay Cao, Lida Guo, Tony Yuen, Jameel Iqbal, Raymond Zhou, Yuanzhen Peng, and Surinder S. Moonga
- Subjects
medicine.medical_specialty ,Anatomy and Physiology ,Bone density ,Ovariectomy ,Osteoporosis ,lcsh:Medicine ,Ascorbic Acid ,Bone resorption ,Mice ,Bone Density ,Internal medicine ,Animals ,Medicine ,lcsh:Science ,Bone ,Biology ,Musculoskeletal System ,Nutrition ,Bone mineral ,Multidisciplinary ,Vitamin C ,biology ,business.industry ,lcsh:R ,Osteoblast ,Vitamins ,medicine.disease ,Ascorbic acid ,Mice, Inbred C57BL ,Radiography ,medicine.anatomical_structure ,Endocrinology ,Osteocalcin ,biology.protein ,lcsh:Q ,Female ,business ,Research Article - Abstract
Epidemiologic studies correlate low vitamin C intake with bone loss. The genetic deletion of enzymes involved in de novo vitamin C synthesis in mice, likewise, causes severe osteoporosis. However, very few studies have evaluated a protective role of this dietary supplement on the skeleton. Here, we show that the ingestion of vitamin C prevents the low-turnover bone loss following ovariectomy in mice. We show that this prevention in areal bone mineral density and micro-CT parameters results from the stimulation of bone formation, demonstrable in vivo by histomorphometry, bone marker measurements, and quantitative PCR. Notably, the reductions in the bone formation rate, plasma osteocalcin levels, and ex vivo osteoblast gene expression 8 weeks post-ovariectomy are all returned to levels of sham-operated controls. The study establishes vitamin C as a skeletal anabolic agent.
- Published
- 2012
34. Blocking antibody to the β-subunit of FSH prevents bone loss by inhibiting bone resorption and stimulating bone synthesis.
- Author
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Ling-Ling Zhu, Blair, Harry, Jay Cao, Tony Yuen, Latif, Rauf, Lida Guo, Tourkova, Irina L., Jianhua Li, Davies, Terry F., Li Sun, Zhuan Bian, Rosen, Clifford, Zallone, Alberta, New, Maria I., and Zaidi, Mone
- Subjects
OSTEOPOROSIS treatment ,MENOPAUSE ,BLOCKING antibodies ,BONE resorption ,ESTROGEN ,BONE growth ,AMINO acid sequence - Abstract
Low estrogen levels undoubtedly underlie menopausal bone thinning. However, rapid and profuse bone loss begins 3 y before the last menstrual period, when serum estrogen is relatively normal. We have shown that the pituitary hormone FSH, the levels of which are high during late perimenopause, directly stimulates bone resorption by osteoclasts. Here, we generated and characterized a polyclonal antibody to a 13-amino-acid-long peptide sequence within the receptor-binding domain of the FSH β-subunit. We show that the FSH antibody binds FSH specifically and blocks its action on osteoclast formation in vitro. When injected into ovariectomized mice, the FSH antibody attenuates bone loss significantly not only by inhibiting bone resorption, but also by stimulating bone formation, a yet uncharacterized action of FSH that we report herein. Mesenchymal cells isolated from mice treated with the FSH antibody show greater osteoblast precursor colony counts, similarly to mesenchymal cells isolated from FSH receptor (FSHR)
-/- mice. This suggests that FSH negatively regulates osteoblast number. We confirm that this action is mediated by signaling-efficient FSHRs present on mesenchymal stem cells. Overall, the data prompt the future development of an FSH-blocking agent as a means of uncoupling bone formation and bone resorption to a therapeutic advantage in humans. [ABSTRACT FROM AUTHOR]- Published
- 2012
- Full Text
- View/download PDF
35. Elevated expression of axin2 and hnkd mRNA provides evidence that Wnt/β-catenin signalling....
- Author
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Dong Yan, Wiesmann, Marion, Rohan, Michael, Chan, Vivien, Jefferson, Ann B., Lida Guo, Sakamoto, Doreen, Caothien, Roger H., Fuller, John H., Reinhard, Christoph, Garcia, Pablo D., Randazzo, Filippo M., Escobedo, Jaime, Fantl, Wendy J., and Williams, Lewis T.
- Subjects
COLON cancer ,NUCLEOTIDES ,MESSENGER RNA ,CELL lines - Abstract
Determines the activation of Wnt/beta-catenin pathway in human colon cancers. Effect of beta-catenin antisense oligodeoxynucleotides on the level of axin2 and human naked cuticle (hnkd) mRNA; Expression of axin2 and hnkd in colon cancer cell lines; Correlation of the increased expression of axin2 and hnkd with truncations in polyposis coli.
- Published
- 2001
- Full Text
- View/download PDF
36. Na+/H+ Exchanger Regulatory Factor 1 (NHERF1) Directly Regulates Osteogenesis.
- Author
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Li Liu, Alonso, Veronica, Lida Guo, Tourkova, Irina, Henderson, Sarah E., Almarza, Alejandro J., Friedman, Peter A., and Blair, Harry C.
- Subjects
- *
BONE growth , *HUMAN growth hormone , *BIOLOGICAL transport , *OSTEOBLAST metabolism , *CELL metabolism , *ACTIVE biological transport - Abstract
Bone formation requires synthesis, secretion, and mineralization of matrix. Deficiencies in these processes produce bone defects. The absence of the PDZ domain protein Na+/H+ exchange regulatory factor 1 (NHERF1) in mice, or its mutation in humans, causes osteomalacia believed to reflect renal phosphate wasting. We show that NHERF1 is expressed by mineralizing osteoblasts and organizes Na+/H+ exchangers (NHEs) and the PTH receptor. NHERF1-null mice display reduced bone formation and wide mineralizing fronts despite elimination of phosphate wasting by dietary supplementation. Bone mass was normal, reflecting coordinated reduction of bone resorption and formation. NHERF1-null bone had decreased strength, consistent with compromised matrix quality. Mesenchymal stem cells from NHERF1-null mice showed limited osteoblast differentiation but enhanced adipocyte differentiation. PTH signaling and Na+/H+ exchange were dysregulated in these cells. Osteoclast differentiation from monocytes was unaffected. Thus, NHERF1 is required for normal osteoblast differentiation and matrix synthesis. In its absence, compensatory mechanisms maintain bone mass, but bone strength is reduced. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
37. Coinfection of Dermacentor silvarum Olenev (Acari: Ixodidae) by Coxiella-Like, Arsenophonus-Like, and Rickettsia-Like Symbionts.
- Author
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Limeng Liu, Lingxia Li, Jiannan Liu, Yonghong Hu, Zhao Liu, Lida Guo, and Jingze Liu
- Subjects
- *
DERMACENTOR , *IXODIDAE , *RNA analysis , *INSECT hosts , *CANDIDATUS , *GENETIC genealogy , *SALIVARY glands - Abstract
We report that multiple symbionts coexist in Dermacentor silva rum. Based on 16S rRNA gene sequence analyses, we prove that Coxiella-llke and Arsenophonus-like symbionts, with 95.6% and 96.7% sequence similarity to symbionts in the closest taxon, respectively, are novel. Moreover, we also provide evidence that the Coxiella-like symbiont appears to be the primary symbiont. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
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