Figure 1.
Morphological inflammation of erythematous and edematous paws and histochemical lesions of synovial hyperplasia and inflammatory infiltration in BIA and CIA mice.
A and D. Control; B and E. BIA mice; C and F. CIA mice. HE staining of sections and microscopic analysis were carried out by sampling hind paws of mice after modeling for 28 d. The histochemical photographs were amplified for 100 folds.
Table 1.
Semi-quantitative evaluation of histological damage by a modified non-parametric scoring system in BIA and CIA mice.
Figure 2.
Morphological inflammation of erythematous and edematous paws and histochemical lesions of synovial hyperplasia and inflammatory infiltration in BIA-CIA mice.
A and B. BIA-CIA mice (intra-dermal CII-CFA injection); C and D. BIA-CIA mice (intra-articular CII-CFA injection). HE staining of sections and microscopic analysis were carried out by sampling hind paws of mice after modeling of BIA-CIA mice (intra-dermal CII-CFA injection) for 28 d and after modeling of BIA-CIA mice (intra-articular CII-CFA injection) for 3 d, respectively. The histochemical photographs were amplified for 100 folds.
Figure 3.
Antibody chip profiling of pro-inflammatory cytokines in blood of BIA, CIA and BIA-CIA mice.
A. BIA mice; B. CIA mice; C. BIA-CIA mice. For protein extraction and antibody hybridization, whole blood was collected from mice after modeling for 28 d, and 40 kinds of cytokines, chemokines and receptors were quantitatively analyzed.
Table 2.
Global modulation of inflammation-related cytokines in blood of BIA, CIA and BIA-CIA mice.
Figure 4.
Dynamic monitoring of NO production and comparative analysis of relationship between NO and SpO2 in BIA, CIA and BIA-CIA mice.
A. Time-course detection of serum NO levels in BIA, CIA and BIA-CIA mice (n = 3). Sampling and detection were conducted every 5 d and until 25 d during modeling; B. Measurement of SpO2 and NO in BIA, CIA and BIA-CIA mice (n = 4). Sampling and detection were conducted after modeling for 3 d in CIA and BIA-CIA mice or after modeling for 28 d in BIA mice. The singular asterisk (*) represents statistically significant difference from the control (P<0.05); and double asterisks (**) indicate statistically very significant difference from the control (P<0.01).
Figure 5.
NO-driven hypoxia and angiogenesis in BIA and CIA mice.
A. Serum NO and LA levels in BIA mice (n = 3); B. Serum NO and LA levels in CIA mice (n = 3); C. Immunohistochemical staining against HIF-1α in the articular synovium of CIA mice (×200); D. Immunohistochemical staining against VEGF in the articular synovium of CIA mice (×200). Sampling and detection were conducted after modeling for 3 d in CIA mice. The singular asterisk (*) represents statistically significant difference from the control (P<0.05); and double asterisks (**) indicate statistically very significant difference from the control (P<0.01).
Table 3.
SNP-induced overexpression of HIF-1α and VEGF in the SNP-injected hypoderm of mice (n = 3).
Figure 6.
Articular inflammatory manifestations in mice after intra-articular injection with SNP, CII-CFA, or combined SNP and CII-CFA.
A. Control; B. 20µ g SNP-injected mice; C. 40 µg CII-CFA-injected mice; D. 20 µg SNP+40 µg CII-CFA-injected mice; E. 4 µg CII-CFA-injected mice; F. 20 µg SNP+4 µg CII-CFA-injected mice. All photographes were taken after injection with SNP, CII-CFA, or combined SNP and CII-CFA for 1 d.
Table 4.
Comparison of SpO2 in articulates of mice after intra-articular injections by SNP, CII-CFA, or SNP+CII-CFA (n = 3).
Figure 7.
Abrogation of NO production after anti-bacteria and/or NOS inhibition in BIA, CIA and BIA-CIA mice.
A. Serum NO levels in drug-administered BIA, CIA and BIA-CIA mice (n = 3). The serum NO level was determined after anti-bacteria and/or NOS inhibition of modeling mice by 60 µg/ml artesunate, 50 µg/ml rapamycin, 15% alcohol or a combination of drugs; B. Serum NO levels in bacteria-fed or antibiotic-administered mice (n = 10). The serum NO level was determined after live bacterial feeding for 7 d and injecting artesunate, cefotaxime, or the combination of artesunate with cefotaxime for 3 d (twice a day). ART: artesunate; CEF: cefotaxime; RAP: rapamycin; ALC: alcohol. The singular asterisk (*) represents statistically significant difference from the control (P<0.05); and double asterisks (**) indicate statistically very significant difference from the control (P<0.01).
Table 5.
Measurement of hypoxic parameters for evaluation of cefotaxime on bacteria fed mice (n = 4).
Figure 8.
Histological amelioration of synovitis by anti-arthritis therapies in CIA and BIA-CIA mice (HE staining, ×200).
A and B. Pre-treatment of CIA (intra-articular CII-CFA injection) mice with 60 µg/ml artesunate or 50 µg/ml rapamycin; C and D. Post-treatment of CIA (intra-dermal CII-CFA injection) mice with 60 µg/ml artesunate and 50 µg/ml rapamycin, or 50 µg/ml rapamycin and 15% alcohol; E and F. Post-treatment of BIA-CIA (intra-dermal CII-CFA injection) mice with 60 µg/ml artesunate and 50 µg/ml rapamycin, or 50 µg/ml rapamycin and 15% alcohol. HE staining of sections and microscopic analysis were carried out by sampling hind paws of mice after therapy completion.