Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits NO, IL-6, and TNF-α production in RAW264.7 and MH-S cells. The cells were treated with JGF (50, 100, 150, 300, 600 μg/mL), 2-E (0.1 μM), DXT (10 μM), or LPS (0.1 μg/mL) for 24 h. ( A ) Cell viability was evaluated using crystal violet. ( B ) NO production was measured using the Griess assay. ( C-D ) IL-6 ( C ) and TNF-α ( D ) levels were determined by ELISA. EC 50 was calculated by CompuSyn software. Data was presented as mean ± standard deviation (SD) for groups (n = 3). Significant differences are denoted as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Griess Assay, Enzyme-linked Immunosorbent Assay, Software, Standard Deviation

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF downregulates 2-E-induced iNOS and COX-2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 24 h. ( A ) Protein levels of iNOS and COX-2 in macrophages were measured by Western blot. ( B-C ) Quantification of iNOS and COX-2 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. The non-detected data showed as – or ND. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of STAT3 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JAK2 and STAT3 were measured by Western blot. ( B-C ) Quantification of phosphorylated JAK2 and STAT3 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of ERK1/2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JNK1/2, ERK1/2, p38, and p65 were measured by Western blot. ( B-C ) Quantification of phosphorylated JNK1/2, ERK1/2, p38, and p65 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF reduces the 2-E-induced proinflammatory cytokines in vivo . ( A ) The experimental scheme for mouse exposure. ( B-F ) Levels of IL-6 ( B ), TNF-α ( C ), IFN-γ ( D ), IL-1β ( E ), and IL-12 ( F ) in lung tissue and serum were measured by ELISA. Data are presented as mean ± SD (n = 9 for serum, except DXT group n = 6; n = 6 for lung tissue, except DXT group n = 3) ( G ) Representative histological images of lung tissue stained with H&E and IHC images for IL-6, TNF-α, and IL-1β expression. ( H-J ) Quantification of IL-6 ( H ), TNF-α ( I ), and IL-1β ( J ) positive areas using ImageJ (n = 3). Significant differences between the control (CTL) group and other groups are denoted by ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Significant differences between the 2-E group and 2-E + JGF group are indicated by #p < 0.05, ##p < 0.01, ###p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Control

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Schematic diagram illustrates the mechanism of Mn-NC composite hydrogels for TMJ-OA treatment. Composite hydrogels can effectively protect chondrocytes from ROS damage, alleviate inflammatory reactions within the joint microenvironment, inhibit inflammation-induced chondrocyte apoptosis and ECM degradation, and simultaneously induce the regeneration and repair of subchondral bones via the enzyme-like activity of Mn-NC SAzymes and the release of Mn ions.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Activity Assay

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Characterization of Mn-NC SAzymes. (a) Preparation of Mn-NC SAzymes. (b) TEM images of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (c) HAADF-STEM image of Mn-NC-10 SAzymes and the corresponding EDX elemental mappings of C, N, O, and Mn. Inset: the SAED pattern. (d) Aberration-corrected HAADF-STEM image of Mn-NC-10 SAzymes. (e) XRD patterns of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (f) Mn 2p and (g) Mn 3s XPS spectra of Mn-NC-10 SAzymes. (h) XANES and (i) FT EXAFS spectra of Mn-NC-10 SAzymes and reference samples at the Mn K-edge. (j) FT EXAFS spectra fitting curves of Mn-NC-10 SAzymes at the Mn K-edge (inset: structure model).

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques:

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: SOD/CAT-like activity and DFT calculations of the Mn-NC SAzymes. The SOD-like activity (a), H 2 O 2 scavenging ratio (b), and dissolved O 2 production curves (c) of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (d) •OH scavenging abilities of Mn-NC-10 evaluated by the electron spin resonance (ESR) spectroscopy. (e) Dissolved oxygen generation catalyzed by Mn-NC-10 at pH 6.6 under varying H 2 O 2 concentrations. (f) Michaelis-Menten kinetic curves of Mn-NC-10 derived from dissolved oxygen production by varying H 2 O 2 concentration. (g) Gibbs free-energy step diagram of the Mn-N 4 . (h) Gibbs free energy step diagram of N 4 C and Mn-N 4 -O in the CAT-like catalytic reaction. (i) Schematic of CAT-like catalytic reaction of N 4 C. (j) Schematic of CAT-like catalytic reaction of Mn-N 4 . (k) Differential charge profiles before and after introducing Mn-O to N 4 C. (l) Differential charge of H 2 O and O 2 before and after adsorption to N 4 C and Mn-N 4 -O (yellow represents gain of electrons, blue represents loss of electrons). TEM images (m), Mn ion releasing amount (n), changes of CAT-like activity (o), and changes of SOD-like activity (p) of Mn-NC-10 SAzymes incubated in PBS (pH = 7.0–7.6) for different time.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Activity Assay, Electron Paramagnetic Resonance, Spectroscopy, Derivative Assay, Concentration Assay, Adsorption, Incubation

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels protect chondrocytes from H 2 O 2 -induced damage by attenuating ROS. (a) Live/dead staining of rat chondrocytes after incubation with different hydrogels for 24 h, scale bar = 200 μm. (b) Cell viability of rat chondrocytes after incubation with different hydrogels for 1, 2, and 3 days. (c) Cell viability, (d) Intracellular ROS content, and (e) Bright-field and ROS staining of rat chondrocytes after incubation with different hydrogels under H 2 O 2 -contaning condition for 24 h, scale bar = 50 μm. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group, #p < 0.05, ##p < 0.01, and ###p < 0.001 suggests a significant difference in comparison to the Control + H 2 O 2 group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Staining, Incubation, Comparison, Control

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels protect chondrocytes from IL-1β-induced damage and promote subchondral bone remodeling by stimulating osteogenic differentiation of human mesenchymal stem cells. Cell viability (a) and expressions of inflammatory factors TNF-α (b), IL-6 (c), and inflammation-related proteins (d) in chondrocytes after stimulation with IL-1β. The expressions of inflammatory factors including TNF-α (e), iNOS (f), IL-6 (g), IL-1β (h) and corresponding proteins (i) from rat macrophages (NR8383) co-cultured with Control, Control + H 2 O 2 , NAC + H 2 O 2 , CH + H 2 O 2 , CH@Mn2+H 2 O 2 groups. (j) ALP-positive area staining images from Control, CH, and CH@Mn2 groups at 7 days and corresponding quantitative ALP activity. Scale bar = 200 μm. (k) Expression of osteogenesis-related genes and proteins including ALP, BMP2, OPN from Control, CH, and CH@Mn2 groups at 7 days. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group, # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the Control + IL-1β, Control + H 2 O 2 or CH group. ▽ p < 0.05, ▽▽ p < 0.01, and ▽▽▽ p < 0.001 suggests a significant difference in comparison to the NAC + IL-1β or NAC + H 2 O 2 group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Cell Culture, Control, Staining, Activity Assay, Expressing, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels inhibit chondrocyte apoptosis and ECM degradation in rats after IL-1β stimulation. Nucleus, apoptosis, and cytoskeleton staining of chondrocytes (a) and flow cytometry analysis (b) after IL-1β stimulation, scale bar = 50 μm. Expressions of apoptosis-related genes and proteins including Bax and Bcl2 (c) and ECM degeneration related genes and proteins including Adamts5 and MMP13 (d) from Control, Control + IL-1β, NAC + IL-1β, CH + IL-1β, and CH@Mn2+IL-1β groups. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the Control + IL-1β group. ▽ p < 0.05, ▽▽ p < 0.01, and ▽▽▽ p < 0.001 suggests a significant difference in comparison to the NAC + IL-1β group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Staining, Flow Cytometry, Control, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 2 weeks. (a) Schematic diagram of in vivo experiments of hydrogel treatment of TMJ-OA. HE staining (b), SO/FG staining (c), immunohistochemical staining of COLII (d) and MMP13 (e), immunofluorescence staining of CD86 (f) and CD206 (g) at the synovial, and immunofluorescence staining of IL-1β (h) in the TMJ of rats after 2 weeks of treatment, scale bar = 100 μm. (i) Histological OARSI score of articular cartilage after 2 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (c–h). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 8 weeks. HE staining (a), SO/FG staining (b), immunohistochemical staining of COLII (c) and MMP13 (d), immunofluorescence staining of CD86 (e) and CD206 (f) at the synovial, and immunofluorescence staining of IL-1β (g) in the TMJ of rats after 8 weeks of treatment, scale bar = 100 μm. (h) Histological OARSI score of articular cartilage after 8 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (b–g). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo subchondral bone repair and regeneration effect of Mn-NC composite hydrogels on TMJ-OA after 2 and 8 weeks. (a, b) The temporomandibular joints of rats were collected at 2 and 8 weeks, three views of the condyles reconstructed by micro-CT scan (coronal, horizontal, and sagittal view), scale bar = 1 mm. Quantitative statistics of trabecular thickness (Tb. Th) (c), trabecular separation (Tb. Sp) (d), and ratios of bone volume (BV/TV) (e) from Sham, MIA, MIA + CH, MIA + CH@Mn2 at 2 and 8 weeks. (f) Trap staining images and quantification analysis after 2 weeks of hydrogels treatment. (g) Trap staining images and quantification analysis after 8 weeks of hydrogels treatment. (h) OPN staining images and quantification analysis after 8 weeks of hydrogels treatment. Data represent means ± SD, n = 3. ∗p < 0.05 and ∗∗p < 0.01 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Micro-CT, Staining, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Underlying mechanism of Mn-NC SAzymes composite hydrogels in treating TMJ-OA. (a) The volcano plot showing the DEGs of IL-1β VS Control. (b) The volcano plot showing the DEGs of CH@Mn2 VS IL-1β. (c) Differential gene clustering thermogram of extracellular matrix organization (gray), regulation of inflammatory response (green) and cell adhesion molecule binding (purple) pathway. (d) Gene Ontology (GO) enrichment analysis of CH@Mn2 VS IL-1β. (e) The KEGG enrichment result of the DEGs of IL-1β VS Control. (f) The KEGG enrichment result of the DEGs of CH@Mn2 VS IL-1β. (g) Differential gene clustering thermogram of the MAPK pathway. (h) Activation of MAPK pathway after IL-1β stimulation. n = 3.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Control, Binding Assay, Activation Assay