Journal: Communications Biology

Article Title: In utero exposure to acetaminophen and ibuprofen leads to intergenerational accelerated reproductive aging in female mice

doi: 10.1038/s42003-019-0552-x

Figure Lengend Snippet: Luteolysis is impaired in 6-month-old F2 ovaries. a FSH and LH secretion in serum samples from 6-month-old ovaries measured by ELISA ( n = 11 control (C) and n = 5 F2 ovaries). b PGF 2α and PGE 2 production in 6-month-old ovaries measured by ELISA ( n = 3 control (C) and n = 5 F2 ovaries). c Relative expression levels of the steroidogenic genes StAR, Cyp11a1, Hsd3β, Lhcgr and 20αHsd in 6-month-old F2 ovaries normalised to 18 S ; ** P < 0.01, **** P < 0.001. d Progesterone secretion was measured by LC/MS in 6-month-old control ( n = 6) and F2 ( n = 5) ovaries; ns not significant. e Proliferation in 2-month-old F1 and F2 and in 6-month-old F2 ovaries was assessed by immunofluorescence using anti-PCNA (green) antibody and Hoechst (HST, blue); CL = corpus luteum; white arrows highlight proliferating CLs and arrowheads indicated stromal cells. f , g Apoptotic cells in 2-month-old F1 and F2 and in 6-month-old F2 ovaries were visualised by TUNEL assay (green) and nuclear staining with Hoechst (HST, blue) ( f ) and apoptotic corpora lutea (CL) were counted ( g ); the data are presented as the percentage of apoptotic CLs. * P < 0.05 ( g ). White and blue arrows indicate apoptotic and non-apoptotic CLs, respectively, and arrowheads indicate growing follicles ( f ). Scale bars = 300 μm. h – j The AKT-PDK1 pathway is activated in 6-month-old F2 ovaries compared with controls. Relative expression levels of Pi3k , Pdk1 , Pten and Mtor normalised to 18 S , in control and F2 ovaries; ** P < 0.01, *** P < 0.005 ( h ). Representative immunoblots of total AKT, phosphorylated AKT (P-AKT) and tubulin ( i ) show that the AKT pathway is activated in F2 ovaries ( n = 3), as indicated by the increased P-AKT/AKT ratio compared with control ( n = 4); *** P < 0.005 ( j )

Article Snippet: The concentration of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in serum was measured with specific EIA ELISA kit (ElabScience Biotech, E-EL-M0511 and E-EL-M0057 respectively, CliniSciences France).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Expressing, Liquid Chromatography with Mass Spectroscopy, Immunofluorescence, TUNEL Assay, Staining, Western Blot

Journal: Bone Research

Article Title: C/EBPβ dictates postmenopausal FSHβ transcription and blockade of AEP/C/EBPβ pathway alleviates osteoporosis

doi: 10.1038/s41413-026-00510-y

Figure Lengend Snippet: #11a but not CF3CN suppresses elevation of FSH expression induced by OVX. a Ovariectomized (OVX) mice fed with vehicle, #11a or CF3CN all displayed hypoplastic thread-like uteri and atrophic ovaries, which were normal in sham mice. Uterine weight was quantified and is presented on the right. b The serum FSH levels of sham and OVX mice fed with vehicle, #11a or CF3CN (left); the serum FSH levels of ovariectomized WT and BDNF -/+ mice treated with vehicle or TrkB agonist R13 (right). n = 4–6 mice per group. c Immunoblotting of the pituitary glands from sham, OVX+ vehicle, OVX + #11a and OVX + CF3CN mice showed that #11a but not CF3CN inhibited FSHβ, pC/EBPβ, C/EBPβ and AEP levels induced by OVX. Data are representatives of three independent experiments. d Quantification of western blotting in ( c ), n = 3 per group. e The FSH levels in pituitary gland from sham and OVX mice treated with vehicle, #11a or CF3CN, detected by ELISA. n = 6 mice per group. f The RT-qPCR results of FSHβ and C/EBPβ mRNA levels in the pituitary glands from sham, OVX+ vehicle, OVX + #11a and OVX + CF3CN mice, n = 3 per group. GAPDH was employed as the internal control. Data represented as mean ± SEM, one-way ANOVA, ns no significance, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Serum mouse follicle-stimulating hormone (FSH) ELISA Kit (catalog#: CSB-E06871m-96T) and mouse luteinizing hormone (LH) ELISA kit (catalog#: CSB-E12770m-96T) were from CUSABIO.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Bone Research

Article Title: C/EBPβ dictates postmenopausal FSHβ transcription and blockade of AEP/C/EBPβ pathway alleviates osteoporosis

doi: 10.1038/s41413-026-00510-y

Figure Lengend Snippet: CF3CN and #11a inhibit osteoclastogenesis. a Representative images (left) and quantification (right) of Tartrate-resistant acid phosphatase-stained (TRAP-stained) sections of the distal femur bone. Scale bar = 500 μm (upper panel), Scale bar = 100 μm (lower panel). Areas of interest used in quantification were randomly selected around the circle-marked regions. Area 1 is located near the distal growth plate, area 2 represents cortical or trabecular bone. n = 3 slice from three mice. b The in-vitro bone resorption assay of RAW 264.7 cells induced by RANK-L with or without treatment of BDNF, 7,8-DHF, CF3CN or #11a for 10 days. Scale bar = 100 μm for toluidine blue staining images (upper). Scale bar = 50 μm for TRAP staining images (lower). c ELISA assays detecting CTX1, a bone resorption marker. Serum levels of CTX1 were measured in sham-operated or OVX mice treated with vehicle, CF3CN, or #11a. CTX1 levels were also assessed in the culture medium of undifferentiated RAW 264.7 cells, as well as in differentiated RAW 264.7 cells treated with vehicle, BDNF, 7,8-DHF, CF3CN, or #11a. d , e Images and quantification of Western blotting of RAW 264.7 cells induced by RANK-L with or without treatment of BDNF, 7,8-DHF, CF3CN or #11a for 4 days. ( n = 3, one-way ANOVA). f AEP enzymatic activity assay of RAW 264.7 cells induced by RANK-L with or without treatment of BDNF, 7,8-DHF, CF3CN or #11a for 4 days. ( n = 4, one-way ANOVA). g Representative images of immunohistochemistry staining of C/EBPβ and AEP of the distal femur bone. Data represented as mean ± SEM, ns no significance, one-way ANOVA, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Serum mouse follicle-stimulating hormone (FSH) ELISA Kit (catalog#: CSB-E06871m-96T) and mouse luteinizing hormone (LH) ELISA kit (catalog#: CSB-E12770m-96T) were from CUSABIO.

Techniques: Staining, In Vitro, Enzyme-linked Immunosorbent Assay, Marker, Western Blot, Enzyme Activity Assay, Immunohistochemistry

Journal: Cells

Article Title: Activation of MT1/MT2 to Protect Testes and Leydig Cells against Cisplatin-Induced Oxidative Stress through the SIRT1/Nrf2 Signaling Pathway

doi: 10.3390/cells11101690

Figure Lengend Snippet: Effects of platinum chemotherapeutic drugs on the levels of gonadotropin, melatonin, and oxidative stress in the peripheral blood of male gastrointestinal tumor patients, and reducing the expression of MT1/MT2 in the testis. ( A ) Human testicular tissue was stained with H&E (scale bar = 50 μm). ( B ) Images of TUNEL staining in human testicular tissue (scale bar = 50 μm). ( C ) Representative immunofluorescent pictures of MT1 in human testicular tissue; 3β-HSD was tagged with red fluorescence and MT1 was tagged with green fluorescence. The nucleus was labeled with DAPI (scale bar = 20 μm). ( D ) Representative immunofluorescent pictures of MT2 in human testicular tissue; 3β-HSD was tagged with red fluorescence and MT2 was tagged with green fluorescence. The nucleus was labeled with DAPI (scale bar = 20 μm). ( E ) MT1 mRNA expression in human testicular tissue. ( F ) MT2 mRNA expression in human testicular tissue. ( G ) GnRH level in human serum. ( H ) FSH level in human serum. ( I ) LH level in human serum. ( J ) Testosterone level in human serum. ( K ) Melatonin level in human serum. ( L ) Content of MDA in human serum. ( M ) SOD activity in human serum. ( N ) Total antioxidant capacity in human serum. Data are presented as the mean ± SEM. N = 3 cases per group in ( A – F ). N = 30 cases per group in ( G – N ). CT, chemotherapy. * p < 0.05, ** p < 0.01 versus the control group or the tumor group.

Article Snippet: A Mouse MT (melatonin) ELISA Kit (Cat#: E-EL-M0788c), Human MT (melatonin) ELISA Kit (Cat#: E-EL-H2016c), GnRH (gonadotropin-releasing hormone) ELISA Kit (Cat#: E-EL-0071c), T (testosterone) ELISA Kit (Cat#: E-EL-0155c), Human FSH (follicle stimulating hormone) ELISA Kit (Cat#: E-EL-H1143c), and Human LH (luteinizing hormone) ELISA Kit (Cat#: E-EL-H6019) were purchased from Elabscience (Wuhan, China).

Techniques: Expressing, Staining, TUNEL Assay, Fluorescence, Labeling, Activity Assay, Control

Journal: Pharmaceutics

Article Title: A Novel Follitropin Analog Inhibits Follitropin Activity In Vitro

doi: 10.3390/pharmaceutics13030325

Figure Lengend Snippet: In vitro activity of FSH variants. ( A ) Biological activities of FSH variants were determined by measurement of induced cAMP in the media. CHO-dhFr cells stably expressing FSHR were exposed to ascending concentrations of FSH variants (0–500 mU/mL) for 20 h. Each curve is the mean ± SE of at least 5 independent experiments ( n ≥ 5), with 3 replicates for each concentration in each experiment. One-way ANOVA showed a dose-dependent increase in activity following exposure to rhFSH-WT ( p < 0.001). Two-way ANOVA indicated significant differences between FSH variants ( p < 0.001). ( B ) Competitive activity of single-chain FSH β deg - α deg vs rhFSH-WT. FSHR-expressing CHO cells were incubated for 20 h with 10 mU/mL of rhFSH-WT and with ascending concentrations of FSH β deg - α deg (0–500 mU/mL). One-way ANOVA indicated significant effects at 200 and 500 mU/mL ( p < 0.001).

Article Snippet: In order to carry out receptor binding and bioactivity assays, CHO/dhFr - cells were stably transfected with the cDNA of human FSH receptor (FSHR) (ORIGINE, Rockville, MD, USA).

Techniques: In Vitro, Activity Assay, Stable Transfection, Expressing, Concentration Assay, Incubation

Journal: Pharmaceutics

Article Title: A Novel Follitropin Analog Inhibits Follitropin Activity In Vitro

doi: 10.3390/pharmaceutics13030325

Figure Lengend Snippet: The primers used for gene amplification in PCR reactions.

Article Snippet: In order to carry out receptor binding and bioactivity assays, CHO/dhFr - cells were stably transfected with the cDNA of human FSH receptor (FSHR) (ORIGINE, Rockville, MD, USA).

Techniques: Amplification

Journal: International Journal of Environmental Research and Public Health

Article Title: Parabens Accelerate Ovarian Dysfunction in a 4-Vinylcyclohexene Diepoxide-Induced Ovarian Failure Model

doi: 10.3390/ijerph14020161

Figure Lengend Snippet: Effect of VCD and parabens on plasma FSH level. Plasma FHS hormone level were measured by ELISA. a p < 0.05 vs. VE. Data are presented as the mean ± SD. VE; 20% Et-OH, VCD 40 mg/kg/day, MP, PP, BP; 100 mg/kg/day.

Article Snippet: A commercially available ELISA kit was used to measure the serum concentration of follicle-stimulating hormone (FSH; ELISA Kit, Cusabio, College Park, MD, USA).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay