Fig 2: Immunohistochemical staining for PAI-1. a Representative photomicrographs of the immunohistochemical analysis of PAI-1 in ectopic and paired eutopic endometria with adenomyosis and control endometrium without adenomyosis. Black arrows indicate the localization of the glandular epithelium (GE), stroma (S), and smooth muscle (SM). b and c Quantitative analysis of the MOD values of PAI-1 immunoreactivity in the glandular epithelium among control, eutopic, and ectopic endometria in the proliferative phase and secretory phase, respectively. d and e Quantitative analysis of the MOD values of PAI-1 immunoreactivity in the stroma among control, eutopic, and ectopic endometria in the proliferative phase and secretory phase, respectively. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. NC, negative control; GE, glandular epithelium; S, stroma; SM, smooth muscle

Fig 3: Therapeutic effects of the active targeting nanotherapy CTLH NP in pregnant rats with stenosis-induced DVT.a Representative digital photos showing thrombi in left iliac veins isolated from DVT pregnant rats after treatment with different formulations. Scale bar, 1 mm. b, c The normalized weight (b) and length (c) of thrombi in left iliac veins after different treatments. d H&E-stained sections of left iliac veins. Scale bar, 400 µm. e Ultrasound imaging of left iliac veins. The yellow dashed lines indicate the vascular endothelium, while the red dashed lines indicate thrombi. Scale bar, 1 mm. f SEM observation of the luminal surface of left iliac veins after different treatments. Scale bar, 10 µm. g–l Relative mRNA levels TAFI, PAI-1, TNF-a, IL-6, NF-?B, and vWF in left iliac veins of different groups. m, n Fluorescence images of DHE-stained cryosections (m) and quantitative analysis of relative ROS levels (n) in left iliac veins. Scale bar, 400 µm. o, p Immunofluorescence analysis of MPO-positive neutrophils in cryosections (o) and quantification of MPO levels (p) in left iliac veins. Scale bar, 400 µm. In all these studies, left iliac veins of pregnant rats at G10 were ligated to induce DVT. At 6 h after the formation of thrombosis, DVT rats were daily administered with saline (the model group), TLH NP (5 mg/kg of LMWH, i.v.), or CTLH NP (5 mg/kg of LMWH, i.v.) for 5 days. In the normal group, healthy pregnant rats with sham operation were treated with saline. Data in a, d–f, m, o are representative of six independent samples. Data in b, c, g–l, n, p are mean ± s.d. (n = 6 independent samples). Statistical significance was assessed by one-way ANOVA with post hoc LSD tests. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

Fig 4: Therapeutic effects of LH NP and TLH NP in pregnant rats with stenosis-induced DVT.a Schematic illustration of the treatment regimens. b Representative digital photos of left iliac veins with thrombi isolated from pregnant rats after treatment with different formulations. Scale bar, 1 mm. c, d The normalized weight (c) and length (d) of thrombi in left iliac veins after different treatments. e H&E-stained histopathological sections of left iliac veins with thrombi. Scale bar, 400 µm. f Ultrasound images of left iliac veins. The yellow dashed lines indicate the vascular endothelium, while the red dashed lines indicate thrombi. Scale bar, 1 mm. g SEM observation of the endothelial surface of left iliac veins with thrombi and after different treatments. Scale bar, 10 µm. h–j Relative mRNA levels of TNF-a, IL-6, NF-?B, PAI-1, TAFI, and vWF in left iliac veins. k, l Fluorescence images of DHE-stained cryosections (k) and quantitative analysis of relative ROS levels (l) in left iliac veins. Scale bar, 400 µm. m, n Immunofluorescence images showing MPO-positive neutrophils in cryosections (m) and quantified MPO levels (n) in left iliac veins. Scale bar, 400 µm. o, p Representative flow cytometric profiles (o) and quantified levels (p) of CD11b+/RP-1+ neutrophils in left iliac veins. In all these studies, left iliac veins of pregnant rats at G10 were ligated to induce DVT. At 6 h after the formation of stenosis-induced thrombi, diseased rats were daily administered with saline (the model group), free Tempol (8 mg/kg, i.v.), free LMWH (5 mg/kg, s.c.), LH NP (at 5 mg/kg of LMWH, i.v.), or TLH NP (at 5 mg/kg of LMWH, i.v.) for 5 days. In the normal group, healthy pregnant rats with sham operation were treated with saline. Data in b, e–g, k, m, o are representative of six independent samples. Data in c, d, h–j, l, n, p are mean ± s.d. (n = 6 independent samples). Statistical significance was assessed by one-way ANOVA with post hoc LSD tests. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

Fig 5: H&E and immunofluorescent staining for uPA (red), uPAR (pink), and PAI-1 (yellow), cell nuclei are stained blue. Magnification × 20 and × 40 on day 10. ME, middle ear; EAM, external acoustic meatus; A, tympanic annulus; single asterisk indicates proliferating epithelium control. Pars tensa of the TM, area around the annulus (H&E). Immunofluorescent staining visible in the thin layer of epithelium covering external surface of TM, mainly in the annular region, and in the epithelium covering neighboring meatus. Staining is also visible in the outer portion of annulus, especially prominent for uPA, and weak for uPAR and PAI-1. Day 1, the area near the annuls with proliferating epithelium, migrating on the surface of TM remnant (H&E). Immunofluorescent staining visible in the proliferating epithelium. The green signal visible on PAI-1 stain presumably comes from sebum autofluorescence. Day 2, the same area as on day 1 with thicker layer of proliferating epithelium migrating on TM remnant (H&E). Immunofluorescent staining visible in the proliferating epithelium. The green signal visible on PAI-1 stain presumably comes from sebum autofluorescence. Day 3, thick layer of proliferating epithelium on the external surface and thinner on the inner surface of TM in the vicinity of annulus (H&E). Immunofluorescent staining visible in the proliferating epithelium on the external surface of TM and in the superficial layer of proliferating epithelium on the inner side. Day 5, the edge of perforation and remnant of fibrous layer of pars tensa surrounded by proliferating squamous epithelium (H&E). Immunofluorescent staining prominent in the layer of proliferating squamous epithelium especially close to the growing front. Weak staining for uPA and uPAR is also present in proliferating fibroblasts. Day 7, the edge of perforation and remnant of fibrous layer of pars tensa surrounded by proliferating fibroblasts and thinner than on day 5 proliferating squamous epithelium on external side of TM. Keratin squames with inflammatory cells on its front (H&E). Immunofluorescent staining visible in the layer of proliferating squamous epithelium and in the thin mucosal layer. Day 10, closed perforation with the layer of squamous epithelium consisting of several layers and loose connective tissue composed of fibroblasts (H&E). Immunofluorescent staining clearly visible in the epithelial layer