Hematoxylin eosin he

Both the liver portion extracted at the time of hepatectomy and that collected at autopsy were processed for histopathological examination. Liver tissues were fixed in 10% formalin for 48 hours, routinely processed, and sliced into sections of 4 μm in thickness. For detection of liver fibrosis, sections were stained with picrosirius red (PSR; Sigma‐Aldrich), anti–α‐smooth muscle actin (α‐SMA) antibodies (AbCam) and anti‐desmin antibodies (Boehringer). For detection of macrophage infiltration, sections were stained with antibodies against CD68 (AbCam) and F4/80 (AbCam). For detection of liver proliferation, sections were stained with anti–proliferating cell nuclear antigen (PCNA) antibodies (Dako). Sections were also stained with hematoxylin‐eosin (H & E) and periodic acid–Schiff (both from Sigma‐Aldrich). After staining, specimens were photographed under a microscope (Olympus). Histological and immunohistochemical results were quantified using WinRoof 7.4 software (Mitani Corporation).

Both left and right kidneys, ureters and bladder were surgically excised and fixed in phosphate buffered saline (PBS) containing 10% formaldehyde and sections were stained with Hematoxylin & Eosin (H&E) (Sigma-Aldrich) by standard protocols as described earlier [13] (link), [14] (link). At least three sections per animal were analyzed by a histopathologist.

New Zealand white rabbits (3–4 weeks; 3.25±0.25 kg) were randomly divided into 4 groups: infection groups (n=30, including Spinal tuberculosis experimental group, Spinal tuberculosis experimental with 20 mg/kg ISL treated group, Spinal tuberculosis experimental with 100 mg/kg ISL treated group, n=10 in each group) and control group (n=10). All of the rabbits were anesthetized with 2% barbital sodium indodine (20 mg/kg; Sigma Aldrich), and holes were drilled on the 6^th lumbar vertebras of rabbits. Gelfoam sponges socked in 0.1 ml H37Rv suspension or normal saline were placed into the perforations of these rabbits. After suturing the incisions, these rabbits were fed alone and 20 mg/kg ISL, 100 mg/kg ISL or normal saline were injected into these rabbits through auricular vein everyday according the groups they belong. The treatment continued for 10 weeks, and tissues surrounding the 5–7 lumbar vertebral bones were dissected for Hematoxylin-Eosin (HE, Sigma Aldrich) staining and peripheral venous bloods were collected for later analyses. All of the experiments were approved by Ethics Committee of Institutional Animal Care and Use Committee at National Defense Medical Center.