Journal: International Journal of Molecular Sciences

Article Title: TRPM8: A Therapeutic Target for Neuroinflammatory Symptoms Induced by Severe Dry Eye Disease

doi: 10.3390/ijms21228756

Figure Lengend Snippet: Mechanical allodynia and corneal epithelial damage in DED mice treated with M8-B (20 µM) vs. PBS. ( A ) Corneal mechanical sensitivity measured with von Frey filaments (g: gram). ( B ) Images and quantification of spontaneous eye closing ratio calculated by measuring the width: height ratio. ( C ) Cornea alteration was examined using a slit-lamp microscope with a cobalt blue filter after placing a drop of fluorescein (0.5% Fluorescein Faure) in the right eye of the animals. The punctuate keratitis (red arrows) was quantified using standard grading scale, 21 days after surgery in DED mice treated with M-8B (20 µM) vs. PBS ( n = 6–9). All tests were conducted at day 21. * p < 0.05 and ** p < 0.01 relative to DED mice treated with PBS. Results are expressed as mean ± SEM. For statistical analysis, the Kolmogorov–Smirnov test was performed followed by a parametric t -test using GraphPad Prism version 7.00 (GraphPad Software, La Jolla, CA, USA).

Article Snippet: An in vivo laser confocal microscope (IVCM, Heidelberg Retina Tomography (HRT) II/Rostock Cornea Module (RCM); Heidelberg Engineering GmbH, Heidelberg, Germany)) was used to examine the entire cornea of anesthetized mice [ ].

Techniques: Microscopy, Software

Journal: The Ocular Surface

Article Title: Diagnostic armamentarium of infectious keratitis: A comprehensive review

doi: 10.1016/j.jtos.2021.11.003

Figure Lengend Snippet: This figure highlights the diagnostic dilemma of infectious keratitis, even in the presence of positive microbiological culture result. This is a case of polymicrobial keratitis which was initially treated for a culture-proven Staphylococcus aureus keratitis with hourly topical antibiotics at day 0. (A – B) At day 4, slit-lamp photograph demonstrating a dense central infiltrate with healing epithelial defect ( yellow arrow ). (C – D) At day 5, slit-lamp photograph demonstrating continued improvement of the ulcer, with gradual contraction of the infiltrate and reduction of epithelial defect ( red arrow). However, the deep-seated infiltrate failed to resolve after 10 days of intensive antibiotic treatment, raising the suspicion of co-existing fungal infection (confirmed on in vivo confocal microscopy). Topical antibiotics was then switched to topical voriconazole 1% hourly. (E – F) At day 24, complete healing of ulcer was achieved with intensive topical voriconazole drops. Deep central pigmented keratic precipitates ( green arrow) and bullous keratopathy secondary to endothelial damage from IK were noted. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Among them, laser scanning IVCM [Heidelberg Retinal Tomograph (HRT), Heidelberg Engineering, GmBH, Germany] [ ] combined with a specially designed, mountable objective system, named Rostock Corneal Module (RCM), has been the preferred choice of IVCM in our practice as well as many others [ ].

Techniques: Diagnostic Assay, Infection, In Vivo, Confocal Microscopy

Journal: The Ocular Surface

Article Title: Diagnostic armamentarium of infectious keratitis: A comprehensive review

doi: 10.1016/j.jtos.2021.11.003

Figure Lengend Snippet: A summary of the recent evidence on the use of polymerase chain reaction (PCR) in the diagnosis of a range of infectious keratitis.

Article Snippet: Among them, laser scanning IVCM [Heidelberg Retinal Tomograph (HRT), Heidelberg Engineering, GmBH, Germany] [ ] combined with a specially designed, mountable objective system, named Rostock Corneal Module (RCM), has been the preferred choice of IVCM in our practice as well as many others [ ].

Techniques: Polymerase Chain Reaction, Biomarker Discovery, Comparison, Multiplex Assay, Touchdown PCR

Journal: The Ocular Surface

Article Title: Diagnostic armamentarium of infectious keratitis: A comprehensive review

doi: 10.1016/j.jtos.2021.11.003

Figure Lengend Snippet: A summary of the diagnostic performance of in vivo confocal microscopy (IVCM) on infectious keratitis (IK).

Article Snippet: Among them, laser scanning IVCM [Heidelberg Retinal Tomograph (HRT), Heidelberg Engineering, GmBH, Germany] [ ] combined with a specially designed, mountable objective system, named Rostock Corneal Module (RCM), has been the preferred choice of IVCM in our practice as well as many others [ ].

Techniques: Diagnostic Assay, In Vivo, Confocal Microscopy

Journal: The Ocular Surface

Article Title: Diagnostic armamentarium of infectious keratitis: A comprehensive review

doi: 10.1016/j.jtos.2021.11.003

Figure Lengend Snippet: (A – B). Various characteristic features of Acanthamoeba cysts on in vivo confocal microscopy (IVCM), including double-wall cysts ( red arrows ), signet rings (yellow arrows ), and bright spots ( green arrows). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Among them, laser scanning IVCM [Heidelberg Retinal Tomograph (HRT), Heidelberg Engineering, GmBH, Germany] [ ] combined with a specially designed, mountable objective system, named Rostock Corneal Module (RCM), has been the preferred choice of IVCM in our practice as well as many others [ ].

Techniques: In Vivo, Confocal Microscopy

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: Preparation of miR140@MVs with autonomous cartilage-penetration ability. (A) The size distribution of miR140@MVs was analyzed NTA. The data are shown as mean ± SEM (red regions), n = 3. (B) TEM images of miR140@MVs. Scale bars: 100 nm. (C) Capillary electrophoresis immunoassay of Anx A1 expression of MVs and miR140@MVs. The gray value of Anx A1 protein expression normalizes to that of MVs analyzed by Image J. (D) The variations in particle size and zeta potential of MVs and miR140@MVs in PBS at 4 °C for 10 days. (E) Schematic illustration of the preparation and the penetration of miR140@MVs into OA-mimic cartilage. (F) The infiltrability of Cy3-miR140@FITC-MVs in OA-mimic cartilage was observed by confocal laser scanning microscopy (CLSM). miR140 was labeled with Cy3 (Red), while MVs were stained by FITC-Annexin V (Green). The cell nuclei were stained with Hoechst 33342 (Blue). Scale bar: 20 μm. The cartilage edge was indicated by a white dotted line. The gray values of FITC-MVs and Cy3-miR140 were analyzed by Image J in line with the white line. The gray arrow indicated the depth of penetration of Cy3-miR140@FITC-MVs. (G) The mean fluorescence intensity (MFI) of Cy3-miR140 in the different regions of cartilages including the shallow region (0–40 μm), the middle region (40–80 μm), and the deep region (80–120 μm). MFI of the shallow region served as a control. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with the Shallow group, n = 3. No significance, ns; ∗ P < 0.05; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: As for subcellular localization analysis, chondrocytes were incubated with Cy3-miR140@DiR-MVs for 6 h, washed with PBS, and sequentially stained with ER-Tracker (1:1000, Beyotime) and LysoTracker (1:2000, Yeasen) for 30 min at 37 °C, followed by Hoechst 33342 (1:2000, Beyotime) nuclear staining for 5 min. Next, cells were washed with PBS for three times.

Techniques: Electrophoresis, Expressing, Zeta Potential Analyzer, Confocal Laser Scanning Microscopy, Labeling, Staining, Fluorescence, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: Rebuilding the metabolic homeostasis of OAC by miR140@MVs. (A) The uptake of Cy3-miR140@FITC-MVs by OAC was observed by using CLSM. miR140, MVs and nuclei were separately labeled by Cy3 (Red), FITC-Annexin V (Green) and Hoechst 33342 (Blue). Scale bar: 5 μm. The gray values were analyzed by Image J followed by the white lines. (B) The distribution of Cy3-miR140@FITC-MVs within OAC was observed by ultra-high resolution confocal microscope (Leica, Stellaris 5) at 6 h co-incubation. ER and lysosomes were labeled by ER-Tracker (Yellow) and Lyso-Tracker (Green). Scale bar: 5 μm. (C) The relative level of miR140 in OAC after incubation with different formulations. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. OAC without treatment served as a control. No significance, ns; ∗∗∗∗ P < 0.0001. (D) Western blotting of COLII, ADAMTS5, MMP13 and FUT1 in OAC treated with miR140@MVs. The gray values of (E) ADAMTS5, (F) MMP13, (G) FUT1 and (H) COLII in (D) were analyzed by Image J and normalized to their respective baseline controls. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Sidak's correction, n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (I) The representative flow cytometry diagram of apoptosis ratios of OAC treated with different formulations. (J) The viability of OAC after different treatments was detected by cell counting kit (CCK)-8. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. OAC served as a control. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: As for subcellular localization analysis, chondrocytes were incubated with Cy3-miR140@DiR-MVs for 6 h, washed with PBS, and sequentially stained with ER-Tracker (1:1000, Beyotime) and LysoTracker (1:2000, Yeasen) for 30 min at 37 °C, followed by Hoechst 33342 (1:2000, Beyotime) nuclear staining for 5 min. Next, cells were washed with PBS for three times.

Techniques: Labeling, Microscopy, Incubation, Control, Western Blot, Flow Cytometry, Cell Counting, CCK-8 Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: miR140@MVs inhibit the pro-inflammatory effect of LPS-induced BMDM in vitro . (A) The intracellular location of Cy3-miR140@FITC-MVs in BMDM cells stimulated with LPS (100 ng/mL) for 24 h (M1 macrophages). miR140 is labeled with Cy3 (Red) and MVs are stained by FITC-Annexin V (Green). The nuclei were stained by Hoechst 33342 (Blue). Scale bar: 5 μm. (B) Representative flow cytometric analysis of BMDM treated with different formulations. The (C) M1 (CD86 + CD206 – ) and (D) M2 (CD86 − CD206 + ) phenotypes were analyzed by Flowjo. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with M1 group. n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗∗ P < 0.0001. (E) Western blotting of TLR4 in LPS-induced BMDM treated with different formulations. (F) The relative expression of the protein band was analyzed by Image J and normalized to its respective baseline controls. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. ∗ P < 0.05. The relative mRNA levels of (G) TNFα , (H) IL6 , (I) IL1β , (J) IL10 , (K) TGFβ in LPS-induced BMDM receiving different formulations. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with the M1 group, n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (L) The TNF α level in LPS-induced BMDM supernatant was detected by ELISA assay. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. No significance, ns; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (F–I) Values are normalized to the M1 (baseline control) group.

Article Snippet: As for subcellular localization analysis, chondrocytes were incubated with Cy3-miR140@DiR-MVs for 6 h, washed with PBS, and sequentially stained with ER-Tracker (1:1000, Beyotime) and LysoTracker (1:2000, Yeasen) for 30 min at 37 °C, followed by Hoechst 33342 (1:2000, Beyotime) nuclear staining for 5 min. Next, cells were washed with PBS for three times.

Techniques: In Vitro, Labeling, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: The deep penetration into cartilage and chondrocyte targeting of miR140@MVs in vivo . (A) Confocal microscopy images of miR140@MVs distribution in cartilage from DMM mice receiving Cy5-miR140@DiR-MVs for 28 days. miR140 was labeled with Cy5 (Red), MVs were stained by DiR (Green), and nuclei were stained by Hoechst 33342 (Blue). Scale bar: 10 μm. The white dotted line showed the cartilage boundary. (B) Orthogonal projections illustrating the colocalization of Cy5-miR140 (red) and DiR-MVs (green) in chondrocytes (blue). Left: the 2.5D imaging of the cartilage; Right: the white dotted line indicated the slice for the colocalization analysis, and the orthogonal projection was used to analyze the section of the x - y axis and y - z axis following the white line. (C) The images of Cy5-miR140@DiR-MVs in cartilage were observed by 3D-SIM. Cartilage was immunostained with COLII (magenta) antibody. Scare bar: 10 μm.

Article Snippet: As for subcellular localization analysis, chondrocytes were incubated with Cy3-miR140@DiR-MVs for 6 h, washed with PBS, and sequentially stained with ER-Tracker (1:1000, Beyotime) and LysoTracker (1:2000, Yeasen) for 30 min at 37 °C, followed by Hoechst 33342 (1:2000, Beyotime) nuclear staining for 5 min. Next, cells were washed with PBS for three times.

Techniques: In Vivo, Confocal Microscopy, Labeling, Staining, Imaging