Journal: Journal of Pharmaceutical Analysis

Article Title: Tailoring a traditional Chinese medicine prescription for complex diseases: A novel multi-targets-directed gradient weighting strategy

doi: 10.1016/j.jpha.2025.101199

Figure Lengend Snippet: Nao Yi Kang decoction (NYKD) inhibits glial cell activation and proliferation. (A, B) Immunofluorescent staining shows that NYKD inhibits astrocyte activation (green) and decreases the number of proliferating astrocytes: representative images of glial fibrillary acidic protein (GFAP) and proliferating cell nuclear antigen (PCNA) double staining (A) and statistic diagram of the number of double positive cells of GFAP and PCNA. (C, D) Immunofluorescent staining shows that NYKD inhibits microglia activation (green) and decreases the number of proliferating microglia: representative images of ionized calcium-binding adaptor molecule 1 (Iba1) and PCNA double staining (C) and statistic diagram of the number of double positive cells of Iba1 and PCNA (D). (E, F) Immunofluorescent staining shows that NYKD decreased the number of M1 microglia: representative images of Iba1 and cluster of differentiation 86 (CD86) double staining (E) and statistic diagram of the number of double positive cells of Iba1 and CD86 (F). (G, H) Immunofluorescent staining shows that NYKD increases the number of M2 microglia: representative images of Iba1 and arginase 1 (Arg1) double staining (G) and statistic diagram of the number of double positive cells of Iba1 and Arg1 (H). Data are presented as mean ± standard deviation (SD) ( n = 5). ∗∗∗ P < 0.001 and ∗∗∗∗ P < 0.0001. ns: not significant. ICH: intracerebral hemorrhage; Medium: medium dose; DAPI: diamidino-phenyl-indole.

Article Snippet: The primary antibodies used were mouse anti-glial fibrillary acidic protein (GFAP) (1:1500, CSB-MA009369A0m; Cusabio Biotech Co., Ltd., Wuhan, China), mouse anti-proliferating cell nuclear antigen (PCNA) (1:1000, 2586; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-ionized calcium-binding adaptor molecule 1 (Iba1) (1:500, 17198; Cell Signaling Technology), rabbit anti-PCNA (1:1000, 13110; Cell Signaling Technology), rabbit anti-cluster of differentiation 86 (CD86) (1:200, 19589; Cell Signaling Technology), and rabbit anti-arginase 1 (Arg1) (1:1000, 93668; Cell Signaling Technology).

Techniques: Activation Assay, Staining, Double Staining, Binding Assay, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization

doi: 10.7150/ijbs.46144

Figure Lengend Snippet: M1 macrophage and STAT1 were excessive in RSA patients. ( A-B ) The dot plot represents labeling of CD14 + CD86 + and CD14 + TNFα + (M1) cells by flow cytometry in decidua of NP subjects (n= 30) and RSA patients (n= 30). ( C ) qRT-PCR analysis of STAT1 expression in the decidua of NP subjects (n= 30) and RSA patients (n= 30). ( D ) STAT1 and p-STAT1 protein levels were measured in decidua of NP subjects (n= 10) and RSA patients (n= 10) by western blot. ( E ) Representative IHC staining images of STAT1 in the decidua of NP and RSA patients (Scale bar, 50 µm, 200×). ( F ) Correlation between p-STAT1 and the proportion of CD14 + CD86 + in decidua of NP subjects (n= 10) and RSA patients (n= 10). ( G ) Correlation between p-STAT1 and the proportion of CD14 + TNF-α + in decidua of NP subjects (n= 10) and RSA patients (n= 10). Values were listed as the mean± SEM. **** P < 0.0001.

Article Snippet: Various fluorescent conjugated antibodies to major histocompatibility complex II (MHCII, 12-5321-82), cluster of differentiation 86 (CD86, 17-0862-81), mouse EGF-like module-containing mucin-like hormone receptor-like 1 (F4/80, 17-4801-82) and C-C motif chemokine receptor 7 (CCR7, 12-1971-82) were from eBioscience (Ben Lomond, CA, USA), cluster of differentiation 80 (CD80, 553769) was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Labeling, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemistry

Journal: International Journal of Biological Sciences

Article Title: MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization

doi: 10.7150/ijbs.46144

Figure Lengend Snippet: miR-103 reduces M1-polarized RAW264.7 macrophages. RAW264.7 cells were transfected with miR-103 mimics/NC or miR-103 inhibitor/INC for 24h, and then stimulated with or without LPS/IFNγ for 24 h. ( A-B ) The mRNA expression of M1 markers CCL2 , CCL5 , CXCL9 , CXCL10 , IL6 , IL12b were determined using qRT-PCR. ( C-D ) The surface levels of CD80, CD86, MHCII, and CCR7 were analyzed by flow cytometry. ( E-F ) Western blot analysed the protein levels of iNOS. Values were listed as the mean± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Various fluorescent conjugated antibodies to major histocompatibility complex II (MHCII, 12-5321-82), cluster of differentiation 86 (CD86, 17-0862-81), mouse EGF-like module-containing mucin-like hormone receptor-like 1 (F4/80, 17-4801-82) and C-C motif chemokine receptor 7 (CCR7, 12-1971-82) were from eBioscience (Ben Lomond, CA, USA), cluster of differentiation 80 (CD80, 553769) was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Flow Cytometry, Western Blot

Journal: International Journal of Biological Sciences

Article Title: MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization

doi: 10.7150/ijbs.46144

Figure Lengend Snippet: miR-103 reduces M1-polarized PM macrophages. PM cells were transfected with miR-103 mimics/NC or miR-103 inhibitor/INC for 24h, and then stimulated with or without LPS/IFNγ for 24 h. ( A-B ) The mRNA expression of M1 markers CCL2 , CCL5 , CXCL9 , CXCL10 , IL6 , IL12b were determined using qRT-PCR. ( C-D ) Surface expression of CD80, CD86, MHCII, and CCR7 were analyzed by flow cytometry, ( E-F ) Western blot analysed the protein levels of iNOS. Values were listed as the mean± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Various fluorescent conjugated antibodies to major histocompatibility complex II (MHCII, 12-5321-82), cluster of differentiation 86 (CD86, 17-0862-81), mouse EGF-like module-containing mucin-like hormone receptor-like 1 (F4/80, 17-4801-82) and C-C motif chemokine receptor 7 (CCR7, 12-1971-82) were from eBioscience (Ben Lomond, CA, USA), cluster of differentiation 80 (CD80, 553769) was from BD Biosciences (Franklin Lakes, NJ, USA).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Flow Cytometry, Western Blot

Journal: Materials Today Bio

Article Title: Hydrogel delivering self-assembled herbal nanoparticles accelerates diabetic wound healing through mitochondrial regulation

doi: 10.1016/j.mtbio.2025.102417

Figure Lengend Snippet: NC@Gel promotes wound repair via the modulation of ROS. ( A ) Immunofluorescence analysis of CD86 (M1 marker) and CD206 (M2 marker). ( B ) Schematic representation of the action of NC NPs. ( C ) Quantitative analysis of CD86 expression in wound tissues (n = 4). ( D ) Quantification of CD206 expression in wound tissues (n = 4). ( E-G ) Immunofluorescence analysis comparing the expression of TNF-α, Nrf2, and NF-κB in wound tissues. ( H-J ) Quantitative analysis of TNF-α, Nrf2, and NF-κB expression in wound tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Antibodies against α-SMA (α-smooth muscle actin, 67735-1-Ig), CD86 (Cluster of differentiation 86, 26903-1-AP), CD206 (Cluster of differentiation 206, 81525-1-AP), Nrf2 (Nuclear factor-erythroid 2-related factor 2, 80593-1), and IL-10 (Interleukin-10, 60269-1-Ig) were purchased from Proteintech (Wuhan, China).

Techniques: Immunofluorescence, Marker, Expressing

Journal: Experimental eye research

Article Title: Mesenchymal stem cell secretome protects against oxidative stress-induced ocular blast visual pathologies

doi: 10.1016/j.exer.2022.108930

Figure Lengend Snippet: List of Taqman assay ID’s for qPCR.

Article Snippet: Gene Assay ID Reference 18S ribosomal RNA ( 18S ) Mm04277571 NR_003278.3 Tumor necrosis factor ( TNFα ) Mm00443258_m1 NM_013693.3 Chemokine (C-C motif) ligand 2 ( CCL2 ) Mm00441242 NM_011333.3 Intercellular cell adhesion molecule 1 ( ICAM-1 ) Mm00516023_m1 NM_010493.2 Interleukin 1 β ( IL1β ) Mm00434228_m1 NM_008361.3 Cluster of Differentiation 86 ( CD86 ) Mm00444543_m1 NM_019388.3 Glial fibric acid protein ( GFAP ) Mm01253033_m1 NM_001131020.1 Glyceraldehydes 3-phosphate dehydrogenase ( GAPDH ) Rn01462662_g1 NM_017008.4 Glial fibric acid protein (Rat GFAP ) Rn01253033_m1 NM_017009.2 Open in a separate window List of Taqman assay ID’s for qPCR.

Techniques: TaqMan Assay

Journal: Journal of the American Heart Association

Article Title: Hepatic Abnormal Secretion of Apolipoprotein C3 Promotes Inflammation in Aortic Dissection

doi: 10.1161/jaha.124.037172

Figure Lengend Snippet: Figure 4. The aortas of patients with AD exhibit abundant M1 macrophages, with significant activation of the TLR2/NLRP3 signaling pathway, and increased release of MMP2 and MMP9. Representative immunofluorescence (A) of aortas from patients with AD (n=4) and normal controls (n=4), with quantification of INOS (B), TLR2 (C), NLRP3 (D), MMP2 (E), and MMP9 (F) expression in patients with AD and normal controls. Representative western blot (G) of aortas from patients with AD (n=8) and normal controls (n=8), with quantification of TLR2 (H), NLRP3 (I), CD86 (J), INOS (K), MMP2 (L), and MMP9 (M) expression in patients with AD and normal controls. Data are presented as mean±SEM. Bars represent the means, and caps represent the SEM. Statistical significance was determined by Mann–Whitney U test in (B) and (C) and by unpaired t test in D through F and H through M. A value of P<0.05 was considered significant. AD indicates aortic dissection; CD, cluster of differentiation; INOS, inducible nitric oxide synthase; MMP, matrix metalloproteinase; NLRP3, NOD-like receptor pyrin domain containing 3; and TLR2, Toll-like receptor 2.

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for 2 hours and then incubated overnight at 4 °C with primary antibodies against apo C3 (1:1000; Affinity; DF8054), TLR2 (1:1000; Proteintech; 17 236- 1- AP), NLRP3 (1:1000; Invitrogen; MA5- 23919), cluster of differentiation 86 (CD86) (1:1000; Proteintech; 13 395- 1- AP), inducible nitric oxide synthase (INOS) (1:1000; Invitrogen; PA1- 036), matrix metalloproteinase (MMP) 2 (1:1000; Proteintech; 10 373- 2- AP), and MMP9 (1:1000; Proteintech; 10 375- 2- AP).

Techniques: Activation Assay, Immunofluorescence, Expressing, Western Blot, MANN-WHITNEY, Dissection