Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: Plasma C1q/tumor necrosis factor-related protein-3 (CTRP3) concentrations in patients without diabetic peripheral neuropathy (non-DPN) and with DPN. Data are presented as means±SD; ***p<0.001 compared with non-DPN.

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: The results of the nerve conduction testing from groups divided according to plasma CTRP3 tertiles

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: BMJ Open Diabetes Research & Care

Article Title: Plasma C1q/tumor necrosis factor-related protein-3 concentrations are associated with diabetic peripheral neuropathy

doi: 10.1136/bmjdrc-2021-002746

Figure Lengend Snippet: Scatter plots showing the correlations of plasma C1q/tumor necrosis factor-related protein-3 (CTRP3) concentrations with nerve conduction parameters. The correlations were assessed by Pearson’s correlation test. M represents motor, S represents sensory, L represents left, R represents right, T represents tibia, P represents peroneal and Su represents sural.

Article Snippet: The plasma CTRP3 concentrations were determined by ELISA with specific antibody against CTRP3 (Proteintech, Wuhan, China) according to standard protocols.

Techniques: Clinical Proteomics

Journal: Redox Biology

Article Title: Melatonin improves cardiac function in a mouse model of heart failure with preserved ejection fraction

doi: 10.1016/j.redox.2018.07.007

Figure Lengend Snippet: Melatonin increased myocardial CTRP3 expression in obese mice. (A-B) Representative blots and results for myocardial CTRP3 expression (n = 6 per group). (C) Representative images of myocardial CTRP3 immunostaining. (D-E) CTRP3 expression in H9c2 cells. H9c2 cells were treated with melatonin for 15 min, after which they were collected for blot detection. Data are expressed as the mean ± SD of six independent experiments (D-E). Differences were compared by one-way ANOVA followed by Tukey's post hoc test. * P < 0.05 compared with the ND+vehicle group, # P < 0.05 compared with the HFD+vehicle group.

Article Snippet: Circulating CTRP3 levels were determined by commercial ELISA kit (Aviva Systems Biology, San Diego, CA) according to manufacturer's instructions.

Techniques: Expressing, Immunostaining

Journal: Redox Biology

Article Title: Melatonin improves cardiac function in a mouse model of heart failure with preserved ejection fraction

doi: 10.1016/j.redox.2018.07.007

Figure Lengend Snippet: Melatonin caused adipocytes to secrete CTRP3 without affecting adiponectin expression or inflammation. (A) CTRP3 expression in 3T3-L1 adipocytes. 3T3-L1 adipocytes were subjected to treatment with melatonin (10 μmol/L) at different time points and then collected for blot detection. (B-C) Representative images and results for CTRP3 immunostaining in adipose tissue after melatonin treatment (n = 5 per group). (D) Circulating CTRP3 levels after melatonin treatment (n = 6 per group). (E) The level of adiponectin in adipose tissues (n = 5 per group). Data are expressed as the mean ± SD of six independent experiments (A). Differences were compared by one-way ANOVA followed by Tukey's post hoc test. * P < 0.05.

Article Snippet: Circulating CTRP3 levels were determined by commercial ELISA kit (Aviva Systems Biology, San Diego, CA) according to manufacturer's instructions.

Techniques: Expressing, Immunostaining

Journal: Redox Biology

Article Title: Melatonin improves cardiac function in a mouse model of heart failure with preserved ejection fraction

doi: 10.1016/j.redox.2018.07.007

Figure Lengend Snippet: Depleting circulating CTRP3 using a CTRP3 antibody largely abolished melatonin-mediated protection against diastolic dysfunction. (A) LVEDP after melatonin treatment (n = 5 per group). (B) -dP/dt with melatonin treatment (n = 5 per group). (C) Alterations in the Tau index after melatonin treatment (n = 5 per group). (D) The level of 4-HNE (n = 5 per group). (E-F) The mRNA levels of Sod2 and Gpx (n = 5 per group). All data are expressed as the mean ± SD. Differences were compared by one-way ANOVA followed by Tukey's post hoc test. * P < 0.05.

Article Snippet: Circulating CTRP3 levels were determined by commercial ELISA kit (Aviva Systems Biology, San Diego, CA) according to manufacturer's instructions.

Techniques:

Journal: Redox Biology

Article Title: Melatonin improves cardiac function in a mouse model of heart failure with preserved ejection fraction

doi: 10.1016/j.redox.2018.07.007

Figure Lengend Snippet: Adipocyte-derived CTRP3 activated Nrf2 to prevent oxidative stress and cell apoptosis. (A) CTRP3 expression in the pooled adipocyte medium. 3T3-L1 adipocytes were treated melatonin (10 μmol/L) or vehicle for 24 h, after which the medium was collected to detect CTRP3 expression. ConM-V: conditioned medium in which adipocytes treated with vehicle were cultured; ConM-M: conditioned medium in which adipocytes treated with melatonin were cultured. (B-C) Nrf2, HO-1 and SOD2 expression in H9c2 cells. H9c2 cells were exposed to ConM-M for different time periods and then collected for blot detection. (D) Immunostaining of Nrf2 in H9c2 cells. H9c2 cells were exposed to ConM-V or ConM-M for 4 h and then collected for blot detection. (E-F) CTRP3 deficiency in adipocytes largely abolished the effects ConM-M on Nrf2 and HO-1 expression in H9c2 cells. (G) Cell viability in all the groups. (H) Sod2 mRNA levels in H9c2 cells. (I) DCFH-DA staining and TUNEL staining. rhCTRP3, recombinant human CTRP3. All data are expressed as the mean ± SD of six independent experiments. Differences were compared by one-way ANOVA followed by Tukey's post hoc test. * P < 0.05.

Article Snippet: Circulating CTRP3 levels were determined by commercial ELISA kit (Aviva Systems Biology, San Diego, CA) according to manufacturer's instructions.

Techniques: Derivative Assay, Expressing, Cell Culture, Immunostaining, Staining, TUNEL Assay, Recombinant

Journal: Redox Biology

Article Title: Melatonin improves cardiac function in a mouse model of heart failure with preserved ejection fraction

doi: 10.1016/j.redox.2018.07.007

Figure Lengend Snippet: miR-200a was closely associated with Nrf2 activation. (A) Nrf2-related miRNA levels. H9c2 cells were exposed to ConM-V or ConM-M for 24 h and then collected for further analysis. (B) miR-93a, miR-193b and miR-200a levels after rhCTRP3 treatment. (C) Keap1 mRNA levels. (D–F) Nrf2 and HO-1 protein levels. rhCTRP3, recombinant human CTRP3. All data are expressed as the mean ± SD of six independent experiments. Differences were compared by one-way ANOVA followed by Tukey's post hoc test. * P < 0.05.

Article Snippet: Circulating CTRP3 levels were determined by commercial ELISA kit (Aviva Systems Biology, San Diego, CA) according to manufacturer's instructions.

Techniques: Activation Assay, Recombinant

Journal: Diabetology & Metabolic Syndrome

Article Title: Serum C1q/TNF-related protein-3 (CTRP3) levels are decreased in obesity and hypertension and are negatively correlated with parameters of insulin resistance

doi: 10.1186/s13098-015-0029-0

Figure Lengend Snippet: The serum CTRP3 levels in different subgroups. A - Comparison of the CTRP3 levels among the four subgroups. B - The serum CTRP3 levels in men and women.

Article Snippet: Serum CTRP3 levels were determined by enzyme-linked immunosorbent assay (human CTRP3 ELISA kit, Aviscera Bioscience, Inc., Santa Clara, CA, USA).

Techniques: Comparison

Journal: Diabetology & Metabolic Syndrome

Article Title: Serum C1q/TNF-related protein-3 (CTRP3) levels are decreased in obesity and hypertension and are negatively correlated with parameters of insulin resistance

doi: 10.1186/s13098-015-0029-0

Figure Lengend Snippet: Linear and multiple regression analysis of the anthropometric and metabolic parameters associated with CTRP3 levels in the subjects

Article Snippet: Serum CTRP3 levels were determined by enzyme-linked immunosorbent assay (human CTRP3 ELISA kit, Aviscera Bioscience, Inc., Santa Clara, CA, USA).

Techniques:

Journal: Diabetes Therapy

Article Title: Astragaloside IV Regulates Insulin Resistance and Inflammatory Response of Adipocytes via Modulating CTRP3 and PI3K/AKT Signaling

doi: 10.1007/s13300-022-01312-1

Figure Lengend Snippet: Actions of AS-IV on the CTRP3 expression in adipocytes. A Effects of AS-IV on CTRP3 mRNA expression in adipocytes. B Actions of AS-IV on CTRP3 protein level in adipocytes. C Effects of CTRP3 siRNA or si-NC transfection on CTRP3 mRNA expression in adipocytes. D Effects of CTRP3 siRNA or si-NC transfection on CTRP3 protein level in adipocytes. N = 3. ** P < 0.01 and *** P < 0.001

Article Snippet: The siRNA that targets CTRP3 (5′- TGGATTTCGUGGUUACCAATT-3′) was synthesized by RiboBio (Guangzhou, China), and siRNA with the scrambled sequence (5′-GTTCAGTTCAGGTTAGTATCT-3′) was used as control siRNA.

Techniques: Expressing, Transfection

Journal: Diabetes Therapy

Article Title: Astragaloside IV Regulates Insulin Resistance and Inflammatory Response of Adipocytes via Modulating CTRP3 and PI3K/AKT Signaling

doi: 10.1007/s13300-022-01312-1

Figure Lengend Snippet: Effects of CTRP3 silencing on the insulin resistance and inflammatory response in adipocytes treated with AS-IV. A Effects of CTRP3 silencing on glucose consumption in AS-IV stimulated AS-IV are shown. B Effects of CTRP3 silencing on IL-6 mRNA expression in AS-IV stimulated AS-IV are shown. C Effects of CTRP3 silencing on TNF-α mRNA expression in AS-IV stimulated AS-IV are shown. D Effects of CTRP3 silencing on IL-6 protein levels in AS-IV stimulated AS-IV are shown. E Effects of CTRP3 silencing on TNF-α protein level in AS-IV stimulated AS-IV are shown. F Effects of CTRP3 silencing on GLUT-4 mRNA expression in AS-IV stimulated AS-IV are shown. * P < 0.05, ** P < 0.01 and *** P < 0.001

Article Snippet: The siRNA that targets CTRP3 (5′- TGGATTTCGUGGUUACCAATT-3′) was synthesized by RiboBio (Guangzhou, China), and siRNA with the scrambled sequence (5′-GTTCAGTTCAGGTTAGTATCT-3′) was used as control siRNA.

Techniques: Expressing

Journal: Diabetes Therapy

Article Title: Astragaloside IV Regulates Insulin Resistance and Inflammatory Response of Adipocytes via Modulating CTRP3 and PI3K/AKT Signaling

doi: 10.1007/s13300-022-01312-1

Figure Lengend Snippet: Effects of CTRP3 silencing on the PI3K/AKT signaling in adipocytes treated with AS-IV. Protein expression levels of p-PI3K, PI3K, p-AKT and AKT are shown. N = 3. * P < 0.05, ** P < 0.01 and *** P < 0.001

Article Snippet: The siRNA that targets CTRP3 (5′- TGGATTTCGUGGUUACCAATT-3′) was synthesized by RiboBio (Guangzhou, China), and siRNA with the scrambled sequence (5′-GTTCAGTTCAGGTTAGTATCT-3′) was used as control siRNA.

Techniques: Expressing

Journal: Diabetes Therapy

Article Title: Astragaloside IV Regulates Insulin Resistance and Inflammatory Response of Adipocytes via Modulating CTRP3 and PI3K/AKT Signaling

doi: 10.1007/s13300-022-01312-1

Figure Lengend Snippet:

Article Snippet: The siRNA that targets CTRP3 (5′- TGGATTTCGUGGUUACCAATT-3′) was synthesized by RiboBio (Guangzhou, China), and siRNA with the scrambled sequence (5′-GTTCAGTTCAGGTTAGTATCT-3′) was used as control siRNA.

Techniques: Concentration Assay, Expressing, Activity Assay, Inhibition

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: Primers used in qRT-PCR .

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: Effects of CTRP3 treatment on brain water content 7 days after ICH. (A) Brain water content in ipsilateral (Ipsi) and contralateral (Contra) hemispheres 7 days after ICH in rats treated with rCTRP3. (B) Brain water content in ipsilateral (Ipsi) and contralateral (Contra) hemispheres 7 days after ICH in rats treated with Lenti-CTRP3. Values are mean ± SEM. n = 8 per group. * p < 0.05 vs. sham; # p < 0.05 vs. vehicle or Lenti.null. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti.CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: Effects of CTRP3 treatment on neurological functions 7 days after ICH. All animals after ICH showed significant neurological deficits based on performance on the modified Garcia (A) , beam balance (B) and wire hanging (C) tests. Rats treated with rCTRP3 showed reduced neurological deficits in all three tests. Treatment with Lenti-CTRP3 starting 2 weeks before induction of ICH showed a tendency to ameliorate neurological deficits (D–F) . Values are mean ± SEM. n = 9 per group. * p < 0.05 vs. sham; ** p < 0.01 vs. sham; # p < 0.05 vs. vehicle or Lenti.null. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti-CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Modification, Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: Effects of CTRP3 treatment on ICH-induced blood-brain barrier (BBB) damage 7 days after ICH. (A) Quantification of Evans blue dye extravasation (blue staining) in the ipsilateral (Ipsi) and contralateral (Contra) hemispheres 7 days after ICH in rats treated with rCTRP3. (B) Quantification of Evans blue dye extravasation (blue staining) in ipsilateral (Ipsi) and contralateral (Contra) hemispheres 7 days after ICH in rats treated with Lenti-CTRP3. Values are mean ± SEM. n = 6 per group. ** p < 0.01 vs. sham; # p < 0.05 vs. vehicle or Lenti.null. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti-CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Staining, Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: CTRP3 encourages angiogenesis in the hematoma border zone 7 days after ICH. (A) Capillary density computed by immunohistochemical staining for CD31 (brown) in the perifocal region in ICH (b) , rCTRP3 (c) and Lenti-CTRP3 (d) 7 days after ICH. A control brain section demonstrates a normal distribution of microvessels in the same region (a) . Bar = 100 μm. (B) Bar graph showing the number of brown stained capillaries 7 days after ICH. Values are mean ± SEM. n = 5 per group. * p < 0.05 vs. sham; ** p < 0.01 vs. sham; # p < 0.05 vs. ICH. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti-CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage; CD31, platelet endothelial cell adhesion molecule-1.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Immunohistochemical staining, Staining, Control, Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: CTRP3 upregulates pAMPK/HIF-1α/vascular endothelial growth factor (VEGF) axis protein expression in the hematoma border zone 7 days after ICH. (A) Western blot analysis of the effect of rCTRP3 on CTRP3, phosphorylated (p) AMP-activated protein kinase (AMPK; Thr172)/AMPK, HIF-1α and VEGF-A. (B–E) Representative ratios of CTRP3, p-AMPK, HIF-1α and VEGF-A protein expression. (F) Western blot analysis of the effect of Lenti-CTRP3 on CTRP3, phosphorylated (p) AMPK (Thr172)/AMPK, HIF-1α and VEGF-A. (G–J) Representative ratios of CTRP3, p-AMPK, HIF-1α and VEGF-A protein expression. Values are mean ± SEM. n = 4 per group. * p < 0.05 vs. sham; ## p < 0.01 vs. vehicle or Lenti.null. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti-CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Expressing, Western Blot, Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: Quantitative RT-PCR analysis of HIF-1α and VEGF-A mRNA levels in the hematoma border zone 7 days after ICH. (A–C) Quantitative RT-PCR analysis of the effect of rCTRP3 on CTRP3, HIF-1α and VEGF-A mRNA levels. (D–F) Quantitative RT-PCR analysis of the effect of rCTRP3 on CTRP3, HIF-1α and VEGF-A mRNA levels. Values are mean ± SEM. n = 4–6 per group. * p < 0.05 vs. sham; ** p < 0.01 vs. sham; ## p < 0.01 vs. vehicle or Lenti.null. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti.CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Quantitative RT-PCR, Recombinant, Over Expression

Journal: Frontiers in Cellular Neuroscience

Article Title: C1q/Tumor Necrosis Factor-Related Protein-3 Attenuates Brain Injury after Intracerebral Hemorrhage via AMPK-Dependent Pathway in Rat

doi: 10.3389/fncel.2016.00237

Figure Lengend Snippet: CTRP3 promotes angiogenesis via an AMPK-dependent signaling mechanism. (A) Western blot analysis of the effects of COM.C and CBO on rCTRP3-induced p-AMPK, HIF-1α and VEGF-A expression. (B–D) Representative ratios of p-AMPK, HIF-1α and VEGF-A protein expression. (E) Modified Garcia test analysis of the effects of COM.C and CBO on neurological function improvement induced by rCTRP3. (F) Evans blue dye extravasation analysis of the effects of COM.C and CBO on ICH-induced BBB damage reduced by rCTRP3. (G) Immunohistochemical staining analysis of the effects of COM.C and CBO on angiogenesis induced by rCTRP3. Capillary density measured by immunohistochemical staining for CD31 (brown) in the perifocal region in ICH (a) , rCTRP3 (b) , COM.C (c) and CBO (d) 7 days after ICH. (H) Bar graph showing the number of brown stained capillaries. Values are mean ± SEM. n = 4–6 per group. ** p < 0.01 vs. ICH; # p < 0.05 vs. rCTRP3; ## p < 0.01 vs. rCTRP3; & p < 0.05 vs. COM.C. CTRP3, C1q/tumor necrosis factor-related protein-3; rCTRP3, recombinant CTRP3; Lenti-CTRP3, Lentivirus overexpression of CTRP3; ICH, intracerebral hemorrhage; COM.C, compound C; CBO, CBO-P11.

Article Snippet: Adult rats were split at random into the following four groups: sham-operated (sham) group, ICH group, ICH + vehicle group and ICH + recombinant CTRP3 (rCTRP3, Chimerigen, USA) group. rCTRP3 was injected intracerebroventricularly (80 μg/kg) 30 min after ICH and then three times per week until the animals were killed.

Techniques: Western Blot, Expressing, Modification, Immunohistochemical staining, Staining, Recombinant, Over Expression