Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Gene Ontology analysis revealed 21 proteins involved in response to wounding.

Article Snippet: In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques:

Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Levels of representative proteins in tears from healthy controls and traumatic vegetative state patients. A List of 7 selected differentially-expressed proteins. Levels of CTSB (B), PRSS1 (C), S100A7 (D), GSTP1 (E), CFH (F), KNG1 (G), and ORM1 (H).

Article Snippet: In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: TOR complex 2 (TORC2) signaling and the ESCRT machinery cooperate in the protection of plasma membrane integrity in yeast

doi: 10.1074/jbc.RA120.013222

Figure Lengend Snippet: ESCRT mutants depend on lipid metabolism and TORC2-Ypk1 signaling for growth and survival. A, GO analysis for cellular processes of 119 genes that showed negative genetic interactions with vps4 Δ . Only significantly enriched GO terms ( p < 0.05) are shown. 15 genes comprised in the GO term lipid metabolic process are shown. Full GO analysis is described in Table S1 . B, equal amounts of vps4 Δ ypk1 Δ double mutants expressing the indicated YPK1 mutant plasmids (or empty vector) in serial dilutions were incubated on auxotrophic selection medium agar plates at 26 °C. C, schematic representation of the activation of the TORC2-Ypk1 signaling pathway in response to membrane stress and its putative interdependence with the ESCRT machinery. Pkh1/2 , yeast orthologues of 3-phosphoinositide-dependent kinase 1. D, equal amounts of the indicated strains in serial dilutions were incubated on auxotrophic selection medium agar plates containing the indicated rapamycin concentrations at 26 °C. All WT strains are the respective vps4 Δ strain re-expressing VPS4 from plasmid. E, growth of the indicated strains in the presence of PalmC. Cells were inoculated to OD 600 nm = 0.1 in auxotrophic selection medium containing the indicated PalmC concentrations and grown for 24 h in 4 independent experiments. The circles indicate the individual measurements. See also Fig. S1 and Table S1 .

Article Snippet: TaqMan gene expression assays were from Thermo Fisher ( YPK1, Sc04141261_s1; ORM1, Sc04125000_s1; ORM2, Sc04149509_s1; housekeeping gene PGK1, Sc04104844_s1).

Techniques: Expressing, Mutagenesis, Plasmid Preparation, Incubation, Selection, Activation Assay, Membrane

Journal: Frontiers in Microbiology

Article Title: Identification of a Novel Serum Biomarker for Tuberculosis Infection in Chinese HIV Patients by iTRAQ-Based Quantitative Proteomics

doi: 10.3389/fmicb.2018.00330

Figure Lengend Snippet: Differentially expressed proteins in the sera of HIV positive patients with active tuberculosis (TB) relative to controls (only HIV positive patients).

Article Snippet: The concentrations of ENG (CUSABIO, CSB-E10030h, China), PSMB2 (CUSABIO, CSB-E17836h, China), HSP90AA1 (CUSABIO, CSB-E13462h, China), HSPA8 (CUSABIO, CSB-EL010829HU, China), CHI3L1 (CUSABIO, CSB-E13608h, China), CAP1 (CUSABIO, CSB-EL004486HU, China), ORM1 (CUSABIO, CSB-EL017237HU, China), RETN (CUSABIO, CSB-E06884h, China), LPA (CUSABIO, CSB-EQ028005HU, China) and ZYX (CUSABIO, CSB-EL027165HU, China) in the serum samples were quantified using ELISA according to the manufacturer’s instructions.

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Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Proteomic analysis of the synovial fluid (SF) samples from patients with different stages of temporomandibular joint osteoarthritis (TMJOA). A) Flow diagram for patient inclusion and grouping. B) Photograph showing joint fluid collection during arthroscopic surgery. C) Flow diagram for the proteomic analysis of SF samples. D) Principal component analysis of SF samples. E) Top 10 genes that showed an upward trend and top 10 genes that showed a downward trend in the three groups. F) Orosomucoid 1 (ORM1) expression in patients with different grades of TMJOA, including Mild (n = 21), Moderate (n = 27), and Severe (n = 15) groups. Data is expressed as mean ± SD and analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's post‐hoc test. G) ORM1 expression in patients with pain (n = 30) or without pain (n = 26). Data is expressed as mean ± SD and analyzed using unpaired Student's t test (two‐tailed). H) ORM1 expression in patients with different degrees of maximum interincisal opening (MIO). Data in MIO ≤ 25 mm (n = 10), MIO 26–35 mm (n = 20), and MIO ≥ 36 mm (n = 29) groups are expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test. I) ORM1 expression in patients with sleep bruxism (n = 26) or without sleep bruxism (n = 23). Data is expressed as mean ± SD and analyzed using unpaired Student's t test (two‐tailed). J) ORM1 concentration in SF samples detected by ELISA assay in Mild (n = 5), Moderate (n = 5), and Severe (n = 5) groups. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test. FOT, fraction of the total; * p < 0.05.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: Expressing, Two Tailed Test, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Histopathological and microcomputed tomography (micro‐CT) analysis showing the effect of ORM1 on maintaining cartilage homeostasis in rats with TMJOA. A) Immunofluorescence analysis of ORM1 in the condylar cartilage of rats with or without TMJOA. Data are expressed as mean ± SD and analyzed using unpaired Student's t test (two‐tailed), n = 3, * p < 0.05. B) Subcellular localization of exogenous ORM1 conjugated with fluorescein isothiocyanate (FITC). C) Hematoxylin–eosin and Safranin O‐fast green staining of the condylar cartilage of rats in various treatment groups. D) Thickness of the condylar cartilage of rats in various treatment groups based on the Hematoxylin–eosin staining. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05 compared with the UAC+NS group. E) Osteoarthritis Research Society International (OARSI) scores of the condylar cartilage of rats in various treatment groups based on the Safranin O‐fast green staining. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05 compared with the UAC+NS group. F) Micro‐CT images of condylar tissue samples from various treatment groups. Comparison of percent bone volume (BV/TV) (G), bone surface/volume ratio (BS/BV) (H), trabecular number (Tb.N) (I), trabecular separation (Tb.Sp) (J), and bone mineral density (BMD) K) among various treatment groups. NS, normal saline; UAC, unilateral anterior crossbite. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05 compared with the UAC+NS group.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: Tomography, Micro-CT, Immunofluorescence, Two Tailed Test, Staining, Comparison, Saline

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Bulk RNA sequencing in C28/I2 cells treated with ORM1, and the inhibitory effect of ORM1 on MMP13 and MMP3. A) Administration on C28/I2 cells and the principal component analysis; B) Volcano map showing the differentially expressed genes (DEGs) of interest; C) Top 20 DEGs rescued after ORM1 treatment; D) Expression change of genes related to osteoarthritis in various treatment groups; E) The transcripts per million (TPM) of MMP13, MMP3, MMP12, and MMP9 in the three groups. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, # p < 0.05 compared with the NC group, * p < 0.05. F) GO analysis of DEGs that were upregulated after IL‐1β treatment and downregulated after ORM1 treatment. G) mRNA expression level of MMP13 and MMP3 in both natural and inflammation environments in C28/I2 and SW1353 cells after treatment with ORM1 protein. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, # p < 0.05 compared with the NC group, * p < 0.05. H) Western blot showing the protein level of MMP13 and MMP3 in both natural and inflammatory environments in C28/I2 and SW1353 cells after treatment with ORM1 protein. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, # p < 0.05 compared with the natural control (NC) group, * p < 0.05. I, J) Immunofluorescence analysis showing the effect of ORM1 on MMP13 and MMP3 in vitro. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, # p < 0.05 compared with the NC group, * p < 0.05. K) Immunohistochemical staining of MMP13 and MMP3 in the condylar cartilage of rats among various treatment groups. L) Comparison of MMP13‐ and MMP3‐ positive cells in rats among various treatment groups. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05 compared with UAC+NS group.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: RNA Sequencing, Expressing, Western Blot, Control, Immunofluorescence, In Vitro, Immunohistochemical staining, Staining, Comparison

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Identification of the interaction between ORM1 and vimentin (VIM). Co‐immunoprecipitation (CoIP) was used to detect binding between ORM1 and VIM in HEK‐293T (A) and C28/I2 cells (B). An immunocolocalization assay was used to examine the colocalization of VIM and exogenous ORM1 (C) or endogenous ORM1 (D) in C28/I2 cells. Data are expressed as mean ± SD and analyzed using unpaired Student's t test (two‐tailed), n = 3, * p < 0.05. Immunocolocalization (E) and quantification (F) of ORM1 and VIM in the condylar cartilage of rats in the three treatment groups, with the yellow lines showing the articular surface and the interface between bone and cartilage. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05. Western blot (G) and immunofluorescence (H) showing the protein level of VIM in both natural and inflammation environments in C28/I2 and SW1353 cells after treatment with ORM1 protein. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05, # p < 0.05 compared with the NC group.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: Immunoprecipitation, Binding Assay, Two Tailed Test, Western Blot, Immunofluorescence

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Inhibitory effect of ORM1 on VIM/MAPK/MMP signaling pathway. A) Effect of ORM1 treatment on the phosphorylation of ERK, JNK, and p38 in C28/I2 and SW1353 cells was detected using western blot. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05, # p < 0.05 compared with the NC group. Expression of MMP13 and MMP3 (B), and the phosphorylation of extracellular signal‐regulated kinase (ERK), c‐Jun N‐terminal kinase (JNK), and p38 mitogen‐activated protein kinase (p38) (C) after overexpression of VIM with or without ORM1 treatment in C28/I2 and SW1353 cells. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05, # p < 0.05 compared with the Myc group. Expression of MMP13 and MMP3 (D), and the phosphorylation of ERK, JNK, and p38 (E) after knockdown of VIM in C28/I2 and SW1353 cells. Data is expressed as mean ± SD and analyzed using unpaired Student's t test (two‐tailed), n = 3, * p < 0.05, # p < 0.05 compared with the NC siRNA group. F) Expression of MMP13 and MMP3 after overexpression of VIM with or without inhibition of ERK, JNK, and p38 in C28/I2 and SW1353 cells. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05. G) The phosphorylation levels of ERK, JNK, and p38 in condylar cartilage of rats were detected by immunofluorescence, with the yellow lines showing the articular surface and the interface between bone and cartilage. H) Quantification of the phosphorylation levels of ERK, JNK, and p38 in condylar cartilage of rats based on the immunofluorescence. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 5, * p < 0.05 compared with the UAC+NS group.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: Phospho-proteomics, Western Blot, Expressing, Over Expression, Knockdown, Two Tailed Test, Inhibition, Immunofluorescence

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: The role of ORM1‐VIM binding on the activities of MAPK pathway and the expression of MMPs. A) Immunocolocalization of ORM1 truncations and VIM in C28/I2 cells. B) CoIP of ORM1 truncations and VIM in C28/I2 cells. C) Expression of MMP13 and MMP3 after overexpression of VIM without or with the ORM1 truncations in C28/I2 and SW1353 cells. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05, # p < 0.05 compared with the Myc+Flag group. D) Phosphorylation of ERK, JNK, and p38 after overexpression of VIM without or with the ORM1 truncations in C28/I2 and SW1353 cells. Data is expressed as mean ± SD and analyzed using one‐way ANOVA followed by Tukey's post‐hoc test, n = 3, * p < 0.05, # p < 0.05 compared with the Myc+Flag group.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: Binding Assay, Expressing, Over Expression, Phospho-proteomics

Journal: Advanced Science

Article Title: Orosomucoid 1 Ameliorates Temporomandibular Joint Osteoarthritis by Maintaining Cartilage Homeostasis

doi: 10.1002/advs.202500028

Figure Lengend Snippet: Schematic diagram of the role of ORM1 in maintaining cartilage homeostasis via suppressing VIM/MAPK/MMP signaling.

Article Snippet: Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the concentration of ORM1 in SF samples by an ELISA kit (KE00137; Proteintech).

Techniques: