Journal: International Journal of Molecular Sciences

Article Title: CHIR99021-Treated Osteocytes with Wnt Activation in 3D-Printed Module Form an Osteogenic Microenvironment for Enhanced Osteogenesis and Vasculogenesis

doi: 10.3390/ijms24066008

Figure Lengend Snippet: Effects of COOME treated by triptonide on osteoblast differentiation. Tript is a Wnt signaling inhibitor. COOME was treated with or without Wnt inhibitor (50 nM) for 24 h. ( A ) qPCR to detect the expression of Wnt target genes in MLO-Y4 cells after treatment. The treated COOME were then co-cultured with ST2 cells for 3 days and then detected by osteogenic differentiation assay. ( B ) AP staining analysis ( C ) AP biochemical quantitative analysis. Scale bar = 100 µm. ( D ) qPCR detection of osteogenic target genes mRNA expression levels. ( E ) Alizarin red S staining. Scale bar = 100 mm. ( F ) Mineralization quantification assay. ( G ) Oil red O staining. Scale bar = 20 µm. ( H ) qPCR detection of mRNA expression of adipogenic differentiation marker genes. Results are expressed as mean ± SD (n = three per group). * indicates p < 0.05, vs. the control group. # indicates p < 0.05, vs. the COOME group.

Article Snippet: Wnt signaling agonist CHIR99021 (C91) and Wnt signaling activator triptonide (Tript) were obtained from MedChemExpress (Shanghai, China), and Lyophilized gelatin methacryloyl (GelMA) and Lithium phenyl (2,6-trimethyl benzoylzoyl) phosphinate (LAP) were obtained from SunP Biotech (Beijing, China).

Techniques: Expressing, Cell Culture, Differentiation Assay, Staining, Marker, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Screening of major hepatotoxic components of Tripterygium wilfordii based on hepatotoxic injury patterns

doi: 10.1186/s12906-023-03836-w

Figure Lengend Snippet: The 12 chemical components of TwHF

Article Snippet: Parthenolide, triptolide, triptonide, triptophenolide, triptobenzene H, hypodiolide, celastrol, demethylzeylasteral, wilforlide A, triptotriterpenic acid A, wilforine, and regelidine (ACMEC Biochemical, Shanghai, Table ) were dissolved in DMSO at a concentration of 50 mM and stored at − 20 °C.

Techniques:

Journal: BMC Complementary Medicine and Therapies

Article Title: Screening of major hepatotoxic components of Tripterygium wilfordii based on hepatotoxic injury patterns

doi: 10.1186/s12906-023-03836-w

Figure Lengend Snippet: Fluorescence area and fluorescence intensity in zebrafish livers

Article Snippet: Parthenolide, triptolide, triptonide, triptophenolide, triptobenzene H, hypodiolide, celastrol, demethylzeylasteral, wilforlide A, triptotriterpenic acid A, wilforine, and regelidine (ACMEC Biochemical, Shanghai, Table ) were dissolved in DMSO at a concentration of 50 mM and stored at − 20 °C.

Techniques: Fluorescence, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Screening of major hepatotoxic components of Tripterygium wilfordii based on hepatotoxic injury patterns

doi: 10.1186/s12906-023-03836-w

Figure Lengend Snippet: IC 50 values of the 12 components in the L-02 and AML-12 cell lines

Article Snippet: Parthenolide, triptolide, triptonide, triptophenolide, triptobenzene H, hypodiolide, celastrol, demethylzeylasteral, wilforlide A, triptotriterpenic acid A, wilforine, and regelidine (ACMEC Biochemical, Shanghai, Table ) were dissolved in DMSO at a concentration of 50 mM and stored at − 20 °C.

Techniques: Concentration Assay

Journal: BMC Complementary Medicine and Therapies

Article Title: Screening of major hepatotoxic components of Tripterygium wilfordii based on hepatotoxic injury patterns

doi: 10.1186/s12906-023-03836-w

Figure Lengend Snippet: Effects of 12 components of TwHF on the mRNA expression levels of TNF-α and IL-1β. Notes: N = 6; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control group; three experimental replicates in all panels. PAR, parthenolide; TRL, triptolide;TRN, triptonide; DEM, demethylzeylasteral; CEL, celastrol; TH, triptobenzene H; WA, wilforlide A; REG, regelidine; TRP, triptophenolide; HYP, hypodiolide; WF, wilforine; TA, triptotriterpenic acid A

Article Snippet: Parthenolide, triptolide, triptonide, triptophenolide, triptobenzene H, hypodiolide, celastrol, demethylzeylasteral, wilforlide A, triptotriterpenic acid A, wilforine, and regelidine (ACMEC Biochemical, Shanghai, Table ) were dissolved in DMSO at a concentration of 50 mM and stored at − 20 °C.

Techniques: Expressing, Control

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: a Representative histology of the testis, epididymis, and sperm from mice that received vehicle (control) and triptonide (treated) at single daily oral doses of 0.8 mg/kg BW for 4 weeks, and those recovered for 4 weeks after 4 weeks of triptonide treatment. Scale bars = 20 μm. Insets represent digitally enlarged framed areas. b Representative images of the gross morphology of sperm (upper panels, scale bars = 10 μm), testis, and epididymis (lower panels, scale bars = 20 mm) from mice that received short-term (27 days) and long-term (3 and 6 months) treatment with triptonide at single daily oral doses of 0.8 or 1.6 mg/kg BW, and those recovered for 45 days after 4 weeks of triptonide treatment. c Micrographs showing spermatozoa from triptonide-treated and control male mice. Bending of sperm heads occurs inside the seminiferous tubules of triptonide-treated male mice, as revealed by scanning electron microscopy (SEM; scale bar = 2 µm). Transmission electron microscopy (TEM) shows residual cytoplasmic contents (*) surrounding the bent sperm head and tail (arrows; scale bar = 1 µm). d Effects of different doses of triptonide (single daily oral gavage for 4 weeks) on sperm morphology in adult C57BL/6J male mice. Individual data points and mean (measure of center) ± SEM (error bars) are shown. ** p < 0.01; *** p < 0.001, as determined by one-way ANOVA with dosing groups ( n = 4) compared to the vehicle control group ( n = 6). e Effects of different doses of triptonide (single daily oral gavage for 4 weeks) on total and forward sperm motility in adult C57BL/6J male mice. Individual data points and mean (measure of center) ± SEM (error bars) are shown. **** p < 0.0001, as determined by one-way ANOVA with dosing groups ( n = 4) compared to the vehicle control group ( n = 6). f – h Effects of different doses of triptonide (single daily oral gavage for 4 weeks) on sperm counts ( f ), relative testis weight (testis weight × 1000/body weight) ( g ), and body weight ( h ) in adult C57BL/6J male mice. Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. No statistical significance (n.s.) among the groups based on one-way ANOVA analyses. i – l Effects of triptonide (single daily oral dose of 0.8 mg/kg BW for 4 weeks) on intratesticular testosterone ( i ), serum testosterone ( j ), FSH ( k ), and LH ( l ) levels in adult C57BL/6J male mice. Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. No statistical significance was detected between the control and treated groups based on one-way analyses of variance (ANOVA) with p < 0.05 considered statistically significant.

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques: Electron Microscopy, Transmission Assay

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: a Representative images showing morphology of sperm from cynomolgus monkeys that received vehicle for 10 weeks (control), before and after triptonide treatment (single daily oral dose of 0.1 mg/kg BW for 7 and 15 weeks), or during recovery (6 and 10 weeks after 8 weeks of triptonide treatment). Scale bars = 5 μm. b – d Effects of short-term (8 weeks) triptonide treatment (single daily oral doses at 0.1 mg/kg BW) on the sperm morphology ( b ), sperm forward motility ( c ), and sperm counts ( d ) in adult male cynomolgus monkeys. Individual data points and mean (measure of center) ± SEM (error bars) are shown. ** p < 0.01; *** p < 0.001, as determined by two-way ANOVA with control ( n = 3) compared to treated ( n = 7) groups. e Representative testicular histology from monkeys that received vehicle (control), before and after triptonide treatment (single daily oral dose of 0.1 mg/kg BW for 8 weeks), or recovered from triptonide treatment (6 weeks after cessation of triptonide treatment). Insets represent digitally amplified, framed areas. Scale bars = 20 μm. f – h Effects of long-term (126 weeks) triptonide treatment (single daily oral doses at 0.1 mg/kg BW) on sperm morphology ( f ), sperm forward motility ( g ), and sperm counts ( h ) in 4 adult cynomolgus male monkeys. i Mating frequency of adult male monkeys that received either vehicle (control) or short-term (8 weeks) and long-term (126 weeks) triptonide treatment (single daily oral dose at 0.1 mg/kg BW). Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. j Fertility performance of adult male monkeys that received vehicle (control) or triptonide (8–126 weeks), and those that recovered from 8 weeks of triptonide treatment (single daily oral dose at 0.1 mg/kg BW).

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques: Amplification

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: a Male mice treated with triptonide (single daily oral doses at 0.8 mg/kg BW) for 4 weeks regained their fertility between days 20 and 25 after cessation of the treatment, as demonstrated by the birth of pups by females mated with previously treated males (single daily oral doses of 0.8 mg/kg BW for 4 weeks). Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. b Litter size and interval of fertility-proven females mated with control male mice or those recovered from 1-month triptonide treatment (single daily oral dose at 0.8 mg/kg BW). Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. No statistical significance between the control and treated groups based on the Kolmogorov–Smirnov t test. c – e Recovery of sperm morphology ( c ), sperm forward motility ( d ), and sperm counts ( e ) in adult male cynomolgus monkeys after 8 weeks of triptonide treatment (a single daily oral dose at 0.1 mg/kg BW). Individual data points and mean (measure of center) ± SEM (error bars) are shown. ** p < 0.05; *** p < 0.01, two-way ANOVA with the recovery group ( n = 4) compared with the control group ( n = 3). The exact p values can be found in the file. f Mating frequency of adult male monkeys that recovered for 8 weeks from triptonide treatment (single daily oral dose at 0.1 mg/kg BW for 8 weeks). Individual data points and mean (measure of center) ± SEM (error bars) are shown. Sample sizes are marked in brackets. g A male baby monkey fathered by an adult male monkey recovered from 8 weeks of triptonide treatment (a single daily oral dose at 0.1 mg/kg BW).

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques:

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: a The EC 50 of triptonide in inhibiting sperm total (TM) and forward (FM) motility in adult male mice. Data points represent mean ± SD; n = 5 for doses 0, 0.4, and 0.8 mg/kg BW, and n = 9 for doses 0.1, 0.125, and 0.2 mg/kg BW. b The EC 50 of triptonide in inducing male infertility in adult mice. Data points represent pregnancy rate, which reflects percentage of pregnancy among all of the plugged females by the males treated with five doses of triptonide (0, 0.1, 0.2, 0.4, and 0.8 mg/kg BW) for 4 weeks. n = 14 for each of the five doses tested.

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques:

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: a Schematics showing a modified drug affinity responsive target stability (DARTS) assay used in this study to identify proteins interacting with triptonide. Protein lysates from total testes of adult male mice treated with triptonide (single daily oral dose at 0.8 mg/kg BW for 4 weeks) were subjected to digestion using different proteinases for various durations. b A representative gel image (upper panel) showing a specific band of ~18 kDa (red frame) unique to the triptonide-treated testes. The bands were cut out for mass spectrometry-based protein identification, and the top five hits were listed in the table (lower panel). c Schematics showing the GlycoLink beads-based affinity purification method used in this study. d A representative gel image showing a specific band of ~60 kDa eluted from triptonide-conjugated beads under acidic and basic conditions. The “input” lane shows the amount of testis lysates used in the assays. The bands were subjected to MS-based protein identification and some of the proteins identified are listed in h . e A representative western blot showing detection of junction plakoglobin/gamma-catenin in the eluates from beads conjugated with or without triptonide, as all we as the input of total testicular lysates used in the assays. f A representative western blot showing detection of keratin 5 in the eluates from beads conjugated with or without triptonide, as all we as the input of total testicular lysates used in the assays. g Schematics showing the immunoprecipitation-based identification of proteins interreacting with the N-terminus of SPEM1. The biotin–streptavidin system was utilized to bind the N-terminus of SPEM1 to magnetic beads. The major proteins identified using this method are listed in h . h A summary of major proteins identified by the three methods used in this study. Junction plakoglobin/gamma-catenin appears to be the most likely drug target (red frame).

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques: Modification, Mass Spectrometry, Affinity Purification, Western Blot, Immunoprecipitation, Magnetic Beads

Journal: Nature Communications

Article Title: Triptonide is a reversible non-hormonal male contraceptive agent in mice and non-human primates

doi: 10.1038/s41467-021-21517-5

Figure Lengend Snippet: Physiologically, SPEM1 interacts with junction plakoglobin through its N-terminus. However, triptonide binds junction plakoglobin with higher affinity, and this binding disrupts normal interactions between junction plakoglobin with SPEM1, causing a phenotype similar to that of Spem1 knockout, i.e., sperm deformation and male infertility.

Article Snippet: In addition to HPLC data provided by the company, the purity of triptonide used was independently validated using NMR elemental analyses in NuMega Resonance Labs, Inc. (San Diego, CA; Supplementary Fig. ).

Techniques: Binding Assay, Knock-Out

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Establishment and Validation of an In Vitro Screening Method for Traditional Chinese Medicine-Induced Nephrotoxicity

doi: 10.1155/2018/2461915

Figure Lengend Snippet: Source of toxic components.

Article Snippet: 4 , Triptonide , Zhongxin Innova, China.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Establishment and Validation of an In Vitro Screening Method for Traditional Chinese Medicine-Induced Nephrotoxicity

doi: 10.1155/2018/2461915

Figure Lengend Snippet: The medicines that were evaluated.

Article Snippet: 4 , Triptonide , Zhongxin Innova, China.

Techniques: