Fig 2: CT images of the patient's abdomen, B-ultrasound and puncture pathology of the subcutaneous nodules of the abdominal wall. A, B show two nodules under the skin of the abdominal wall, as shown by the yellow and red arrows. The blue arrow shows the subcutaneous strip of high-density shadows, suggesting inflammation. C Shows enlarged spleen. D is a B-ultrasonic image of abdominal wall nodules. There is a 37 mm × 20 mm, 24 mm × 14 mm mixed echo group in the fat layer of the left abdominal wall, with irregular shape, uneven internal echo, and insignificant internal blood flow signal. Under ultrasound-guided puncture of the left abdominal wall nodule, the pathological image is shown in E (Hematoxylin and eosin staining of the nodule, original magnification × 200. Scale bar = 50 µm). Microscopic images were taken with VS200 Slide Scanner (Olympus, Tokyo, Japan) at a resolution of 1880 × 1048 pixels, analyzed with Olyvia software (Olympus). A large number of inflammatory cells including lymphocytes and neutrophils are seen, as well as some pus cells which are fragments of inflammatory cells. A few fibroblasts can be seen among inflammatory cells. Colony smear showed filamentous fragment forms of Acid fast bacilli by Acid-fast stain in F (Original magnification × 1000. Scale bar = 5.0 µm). Microscopic images were taken with VS200 Slide Scanner (Olympus, Tokyo, Japan) at a resolution of 1560 × 920 pixels, analyzed with Olyvia software (Olympus)

Fig 3: Pathological findings in the brain (a, b) and upper respiratory tract (c). a Inferior view of the brain. No macroscopic abnormalities were observed. b Hematoxylin and eosin staining of the brain. Some perivascular extravasation of serum proteins and dilatation of Virchow Robin spaces are found without inflammatory cell infiltration. Original magnification x100. Scale bar = 200 µm. Microscopic images were taken with VS200 Slide Scanner (Olympus, Tokyo, Japan) at 0.274 µm/pixel, and analyzed with Olyvia software (Olympus). The image was not edited. b Upper respiratory tract. Redness was observed on the palatine tonsils (arrow), lingual tonsils (*), and the entire tracheal and bronchial mucosa (arrowhead)

Fig 4: Representative case 2 with a target lesion in a previously difficult-to-reach area. A Computed tomography image revealing a pulmonary nodule 21.8 mm in diameter in the left upper segment 1 + 2. B The location of the lesion is confirmed by radial endobronchial ultrasound (R-EBUS), within the lesion (arrow), under X-ray fluoroscopic guidance. C X-ray fluoroscopic image showing the 1.7-mm cryoprobe reaching the lesion as smoothly as the R-EBUS probe because of its flexibility. D Hematoxylin and eosin staining of the specimen (magnification, × 10). E Higher magnification (square) revealing adenocarcinoma in which tumour cells proliferate while forming glandular structures (magnification, × 100). The stained tissue sections were scanned with 40 × objective using a high-resolution digital slide scanner (SLIDEVIEW VS200, Olympus, Tokyo, Japan) and viewed using VS200 ASW (version 3.1, Olympus). All images were scanned at 24-bit true colour and image manipulation was not performed

Fig 5: Effects of early-life stress on cognition, hippocampal neurogenesis, and neurotrophin levels in the dentate gyrus in adult mice.A The experimental timeline of the behavioral procedure and brain tissue acquisition after stress exposure. B In the spatial object recognition task, control mice distinguished the displaced object from the non-displaced one, whereas stressed mice failed to do so and performed worse than the controls. C ELS did not affect the performance in the Y-maze spontaneous alternation test. The number of DCX-positive (D) and Ki-67-positive (E) neurons in the DG (arrowheads) was decreased in the stressed mice. Scale bar = 100 µm or 10 µm. Immunostaining revealed that ELS significantly reduced the protein levels of BDNF (F), not NT-3 (G) or NGF (H), in the DG in adult mice. Representative images in (D–H) were captured using the Olympus VS200 virtual slide scanning system. Arrowheads, positive neurons; scale bar = 100 µm or 30 µm. AAR alternate arm return, Acc. acclimation, BDNF brain-derived neurotrophic factor, CT control, DCX doublecortin, DG dentate gyrus, ELS early-life stress, gcl granule cell layer, ml molecular layer, NGF neurotrophin nerve growth factor, NT-3 neurotrophin 3, OF open field, PND postnatal day, SA spontaneous alternation, SAR same arm return, SOR spatial object recognition. *p < 0.05, unpaired t test; ###p < 0.001, one-sample t test; &&&p < 0.001, the main effect of stress.