Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the MMP‐2 levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M)+ ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. MMP‐2 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; MMP‐2, matrix metalloproteinase‐2; P 4 , progesterone.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the TNF‐α and IL‐6 levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M) + ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. (A) represents subsequent time points for TNF‐α analysis, and (B) represents subsequent time points for IL‐6 analysis. TNF‐α and IL‐6 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; IL‐6: interleukin‐6; MMP‐2, matrix metalloproteinase‐2; P 4 , progesterone; TNF‐α, tumor necrosis factor‐alpha.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the VEGF‐A levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M)+ ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. VEGF‐A levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; P 4 , progesterone; VEGF‐A, vascular endothelial growth factor‐A.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the IGFBP‐1 secretions in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M) + ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser+ E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser + E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. IGFBP‐1 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; IGFBP‐1, insulin‐like growth factor‐binding protein‐1; P 4 , progesterone.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification, Binding Assay

Journal: Cancers

Article Title: Intermittent Hypoxia Mediates Paraspeckle Protein-1 Upregulation in Sleep Apnea.

doi: 10.3390/cancers13153888

Figure Lengend Snippet: Figure 1. PSPC1 expression and its cleavage in patients with OSA. (a) The mean fluorescence intensity (MFI) of the monocyte PSPC1 intracellular expression (as determined by flow cytometry) in HV (n = 8) and the patients with severe OSA (n = 15) are shown. The PSPC1 distribution (left panel) and PSPC1 histogram (right panel) are shown; (b) The monocytes PSPC1 mRNA expression (estimated by qPCR) in the monocytes from HV (n = 23) and the patients with severe OSA (n = 50) are shown; (c) The PSPC1 plasma protein was quantified using ELISA (n = 18 HV and n = 45 patients with severe OSA). The intergroup differences in PSPC1 expression were assessed using an unpaired t-test with Welch’s test correction. Error bars: Standard error of the mean (SEM). * p < 0.05, ** p < 0.01 when compared with HV; (d) Correlation between PSPC1 and TGFβ mRNA expression in the monocytes from the patients with severe OSA (n = 45) (left panel) and the correlation between PSPC1 and TGFβ protein concentration in the plasma of the patients with severe OSA (n = 40) (right panel). The patients were randomly selected. Spearman correlation coefficients (r) and p-values are shown; (e) Correlation between MMP2 and PSPC1 mRNA expression in the monocytes from the patients with severe OSA (n = 45) (left panel) and the correlation between MMP2 and PSPC1 protein concentration in the plasma of the patients with severe OSA (n = 40) (right panel). Pearson’s correlation coefficients (r) and p-values are shown; (f) MMP2 mRNA expression analysis by qPCR in the monocytes from HV (n = 18) and the patients with severe OSA (n = 45) (left panel). The MMP2 protein was quantified using ELISA (n = 20 HV, and n = 40 patients with OSA) (right panel). The groups were compared using an unpaired t-test with Welch’s test correction. Error bars: SEM. * p < 0.05, **** p < 0.0001 as compared with HV; (g) The PSPC1 expression by flow cytometry analysis is shown. Monocytes from HV (n = 3) were cultured under normoxia or IH for 24 h and then treated with MMP2 for 3 h (left and middle panels). Supernatants were collected to evaluate PSPC1 protein concentrations using ELISA (right panel). Mean values are shown. Spearman coefficients (r) and p-values (p) were calculated for the left panel N (r = 0.3143, p = 0.5639) and IH (r = −0.02, p = >0.99) and right panel N (r = −0.8286, p = 0.0583) and IH (r = −1.00, p = 0.0028); (h) Western blot analysis of enzymatic digestion of PSPC1 with or without MMP2 (molar ratio of 1:10) is shown (left panel). The expected size of 59 kDa (uncleaved) and an additional band of approximately 43 kDa (cleaved) were observed. Three different experiments were performed with similar findings (right panel). The comparison was performed by unpaired t-test; ** p < 0.01.

Article Snippet: Recombinant human MMP2 (R&D Systems) and PSPC1 were dissolved in 20-mM Tris buffer containing 2-mM CaCl2 and 1-mM MgCl2 (pH 7.3), at a concentration of 10 μg/100 L and 100 μg/100 μL, respectively.

Techniques: Expressing, Cytometry, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Protein Concentration, Cell Culture, Western Blot, Comparison

Journal: Cancers

Article Title: Intermittent Hypoxia Mediates Paraspeckle Protein-1 Upregulation in Sleep Apnea.

doi: 10.3390/cancers13153888

Figure Lengend Snippet: Figure 2. HIF1α is associated with PSPC1. (a) Correlation between PSPC1 and HIF1α mRNA expression in monocytes in patients with OSA (n = 40). Pearson correlation coefficients (r) and p-values are shown; (b) Monocytes from HV (n = 7) were treated with 30 µM of PX478 and/or exposed to IH for 16 h. Paired control samples were incubated under normoxia conditions and without PX478. The distribution estimation of MFI of the PSPC1 intracellular expression (as determined by flow cytometry, left panel) and the PSPC1 histogram (right panel) are shown; (c) PSPC1 mRNA expression estimated by qPCR in the monocytes from HV (n = 7) treated with a specific HIF1α inhibitor (30 µM of PX478) or not and exposed to IH or normoxia conditions for 16 h (left panel), monocytes from HV (n = 3) treated with or without siHIF1α and exposed to IH or normoxia conditions for 16 h (middle panel), monocytes from HV (n = 3) treated or not with DMOG for 2 h (right panel); (d) Correlation between MMP2 and HIF1α mRNA expression in the monocytes from the patients with severe OSA (n = 40). Spearman’s correlation coefficients (r) and p-values are shown; (e) MMP2 mRNA expression estimated by qPCR in the monocytes from HV (n = 7) treated with a specific HIF1α inhibitor (30 µM of PX478) or not and exposed to IH or normoxia conditions for 16 h (left panel), monocytes from HV (n = 3) treated with or without siHIF1α and exposed to IH or normoxia conditions for 16 h (middle panel), monocytes from HV (n = 3) treated or not with DMOG for 2 h (right panel). The groups were compared with a Two-way ANOVA or paired t-test. Error bars: SEM. * p < 0.05; ** p < 0.01, *** p < 0.001, compared with the untreated cells. Spearman’s correlation coefficients (r) and p-values are shown.

Article Snippet: Recombinant human MMP2 (R&D Systems) and PSPC1 were dissolved in 20-mM Tris buffer containing 2-mM CaCl2 and 1-mM MgCl2 (pH 7.3), at a concentration of 10 μg/100 L and 100 μg/100 μL, respectively.

Techniques: Expressing, Control, Incubation, Cytometry

Journal: Cancers

Article Title: Intermittent Hypoxia Mediates Paraspeckle Protein-1 Upregulation in Sleep Apnea.

doi: 10.3390/cancers13153888

Figure Lengend Snippet: Figure 6. PSPC1 expression in patients with OSA increases TGFβ expression effect on EMT-TFs. Intermittent hypoxia increases the activation of HIF1α, which might bind with the promoter of genes such as PSPC1, MMP2, and TGFβ, leading to high levels of these proteins in severe OSA monocytes. The MMP2 plays a role in PSPC1 cleavage, increasing plasma PSPC1 levels. The combination of high plasma levels of TGFβ and PSPC1 increases the EMT-TF effect through TWIST and SLUG and CSC effect by SOX2 in melanoma cells.

Article Snippet: Recombinant human MMP2 (R&D Systems) and PSPC1 were dissolved in 20-mM Tris buffer containing 2-mM CaCl2 and 1-mM MgCl2 (pH 7.3), at a concentration of 10 μg/100 L and 100 μg/100 μL, respectively.

Techniques: Expressing, Activation Assay, Clinical Proteomics