Cryo-electron tomography (cryo-ET) enables the determination of high-resolution three-dimensional structures of macromolecular complexes within cells in a near-physiological state, providing crucial structural insights into fundamental life processes. Cryo-ET has achieved landmark successes in single-cell models. However, many critical biological processes do not occur in isolated cells but emerge from intercellular coordination within tissues. Furthermore, many research subjects, including neural tissues, tumor biopsies, plant tissues, and clinical pathological samples, cannot be obtained through single-cell culture and must be directly dissected from organisms or tissue blocks. Advancing cryo-ET from single-cell to tissue-level applications is therefore crucial for capturing the full complexity of biological activities in their native context. A major technical bottleneck for tissue cryo-ET lies in the preparation of sufficiently thin (<300 nm) lamellae from vitrified tissue specimens. Although high-pressure freezing can vitrify tissues up to 200 µm thick, these samples are far too thick for direct transmission electron microscopy imaging. Among the available thinning methods, cryo-focused ion beam (cryo-FIB) milling has emerged as the most promising approach, as it avoids the mechanical artifacts inherent to cryo-ultramicrotomy. However, conventional on-the-grid cryo-FIB milling is inefficient for thick tissues, requiring excessive milling time and discarding most of the sample. To overcome these limitations, cryo-lift-out has been developed—a technique in which a micromanipulator physically extracts a chunk of interest from deep within the tissue and transfers it to a dedicated grid for final thinning. This approach bypasses the thickness barrier and enables site-specific analysis of internal structures. This review systematically traces the evolution of cryo-lift-out from its origins in materials science to its adaptation for biological tissues. In room-temperature lift-out, reliable attachment is achieved by gas-injection system (GIS)-assisted metal deposition. Transferring this approach to cryogenic conditions proved challenging because precursor gases condense on all cold surfaces, leading to contamination and poor adhesion. The development of copper-assisted redeposition marked a critical turning point: instead of relying on gas deposition, this method uses ion-beam sputtering to deposit copper atoms at the needle-chunk interface, creating a strong, low-contamination bond. This innovation has enabled robust cryo-lift-out workflows and paved the way for serial lift-out, in which multiple consecutive lamellae are prepared from a single tissue chunk, substantially increasing throughput and enabling volumetric imaging. Despite these advances, several technical challenges remain. Curtaining effects caused by uneven chunk surfaces can introduce artifacts into tomograms, requiring careful optimization of milling parameters and protective coating. The cryo-adhesion step still demands precise control of beam angle, needle positioning, and milling depth, making the process highly operator-dependent. Additionally, the choice of grid geometry is critical. Custom-designed grids with double-sided attachment improves stability and offer better compatibility with cryo-ET tilt series. Automation, which has greatly improved room-temperature lift-out, has not yet been achieved for cryo-lift-out due to the complexity of handling heterogeneous biological tissues and the need for real-time adaptation. Future progress will likely focus on integrating cryo-lift-out with volume electron microscopy to correlate ultrastructure across scales, developing intelligent control systems to reduce user intervention, and extending the technology to challenging samples such as plant tissues and some material science samples for interface study. A systematic analysis of the cryo-lift-out technique clarifies the key limiting factors for its large-scale application and lays a foundation for methodological refinement and technological innovation. By consolidating recent advances and identifying remaining bottlenecks, this review aims to support the broader adoption of cryo-lift-out and accelerate the development of tissue-scale in situ structural biology.

This study reports a case of a 56-year-old female patient with BRAF-mutated non-small cell lung cancer (NSCLC) who successfully underwent radical surgery after neoadjuvant targeted therapy with the BRAF inhibitor dabrafenib combined with the MEK inhibitor trametinib. The chest drainage tube was removed 2 days postoperatively, and the patient was discharged smoothly. Postoperative pathology indicated invasive adenocarcinoma, moderately to highly differentiated, with 80% being lepidic type, and the maximum tumor diameter was 4 cm. No vascular tumor thrombus, vascular invasion, nerve invasion, spread through air spaces, pleural invasion, or lymph node metastasis were observed. The postoperative staging was ypT2aN0M0. The patient continued with adjuvant treatment with dabrafenib combined with trametinib postoperatively, and no signs of recurrence were found in the follow-up examination six months after surgery.

[Objective] To evaluate the efficacy and safety of sacral neuromodulation (SNM) in the treatment of patients with benign prostatic hyperplasia (BPH) complicated with underactive bladder (UAB) who respond poorly to transurethral resection of the prostate (TURP). [Methods] A retrospective analysis was performed on 10 patients with BPH and UAB treated with TURP by the same surgeon in Zhongda Hospital Southeast University during Jan.2018 and Jan.2023.The residual urine volume was not significantly relieved after operation, and the maximum urine flow rate and urine volume per discharge were not significantly improved.All patients underwent phase I SNM, and urinary diaries were recorded before and after surgery to observe the average daily frequency of urination, volume per urination, maximum urine flow rate, and residual urine volume. [Results] The operation time was (97.6±11.2) min.During the postoperative test of 2-4 weeks, if the residual urine volume reduction by more than 50% was deemed as effective, SNM was effective in 6 patients (60.0%). Compared with preoperative results, the daily frequency of urination [(20.2±3.8) times vs. (13.2±3.2) times], volume per urination [(119.2±56.7) mL vs. (246.5±59.2) mL], maximum urine flow rate [(8.7±1.5) mL/s vs. (16.5±2.6) mL/s], and residual urine volume [(222.5±55.0) mL vs. (80.8±16.0) mL] were significantly improved, with statistical significance (P<0.05). There were no complications such as bleeding, infection, fever or pain.The 6 patients who had effective outcomes successfully completed phase II surgery, and the fistula was removed.During the follow-up of 1 year, the curative effect was stable, and there were no complications such as electrode displacement, incision infection, or pain in the irritation sites.The residual urine volume of the other 4 unsuccessful patients did not improve significantly, and the electrodes were removed and the vesicostomy tube was retained. [Conclusion] SNM is safe and effective in the treatment of BPH with UAB patients with poor curative effects after TURP.

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.

Objective:To investigate the causes and management strategies of anesthetic complications during percutaneous spinal endoscopic surgery under local anesthesia.Methods:A total of 16 800 patients (8 625 males and 8 175 females) who underwent percutaneous spinal endoscopic surgery under local anesthesia (including intravenous basic anesthesia) in Tianjin Hospital, Shandong Public Health Clinical Center and Hebei General Hospital from February 2012 to February 2023 were retrospectively analyzed. The average age was 45.3±21.6 years (range, 12-84 years). There were 220 cases of posterior cervical keyhole endoscopic surgery, 50 cases of thoracic transforaminal endoscopic surgery, 70 cases of thoracic posterior interlaminar endoscopic surgery, 11 670 cases of lumbar transforaminal endoscopic surgery, and 4 790 cases of lumbar posterior interlaminar endoscopic surgery. The occurrence time, clinical manifestations, management of intraoperative anesthesia complications were recorded, as well as surgical segments, puncture sites, complication symptoms, signs, outcome and prognosis.Results:All patients received percutaneous water-mediated uniaxial spinal endoscopic surgery under local anesthesia. There were 9 patients experienced anesthesia complications, including 6 cases of epidural diffusion of anesthetics and 3 cases of anesthetics mistakenly entering the subarachnoid space. There were 4 males and 5 females, aged 48.4±18.2 years (range, 28-84 years). There were 1 case of T 12L 1 disc herniation, 1 case of C 5-6 disc herniation, 3 cases of L 4-5 disc herniation and 4 cases of L 5S 1 disc herniation. Surgical segments and procedures: 1 case of C 5-6 posterior Keyhole endoscopic surgery, 1 case of T 12L 1 transforaminal endoscopic surgery, 2 cases of L 4-5 transforaminal endoscopic surgery, 1 case of L 4-5 interlaminar endoscopic surgery, and 4 cases of L 5S 1 interlaminar endoscopic surgery. Anesthesia complications all appeared 5-10 min after injection of local anesthetics, with symptoms of decreased oxygen saturation, decreased blood pressure, altered consciousness, and sensory and motor dysfunction of limbs. 6 patients with epidural diffusion of anesthetics recovered completely after symptomatic treatment in 5 cases, and 1 case was left with foot drop. Three patients with anesthetics mistakenly entering the subarachnoid space were immediately converted to the supine position, of which one recovered by mask oxygenation; 1 patient improved after emergency tracheal intubation, rehydration, and application of vasoconstrictive medications; and 1 patient developed multiple complications such as multiorgan failure, rhabdomyolysis, and sepsis after tracheal intubation, and recovered at 3 months after surgery with symptomatic treatment. Conclusions:Epidural diffusion and entering into subarachnoid space of anesthetics are serious complications of local anesthesia in percutaneous spinal endoscopic surgery. In addition to sensory and motor dysfunction of the limbs, the functions of the respiratory and circulatory systems can also be affected. It is necessary to be alert to the occurrence of anesthesia-related complications during operation and early identification and treatment.

Objective:To investigate the inpact of thyroid cancer-derived cancer-associated fibroblasts(CAF) autophagy on papillary thyroid cancer(PTC).Methods:CAF and normal fibroblasts were isolated from cancerous and adjacent normal thyroid tissues from four PTC patients. Expressions of fibroblast activation protein(FAP) and α-smooth muscle actin in cells were assessed. Conditioned medium of CAF and normal fibroblasts were prepared and used to culture PTC cells. The effects of CAF and normal fibroblasts on survival, proliferation, migration, invasion and iodine uptake of PTC cells were evaluated through cell proliferation assay, cell scratch assay, cell invasion assay, and cell iodine uptake assay. The autophagy level of CAF was also evaluated. Autophagy inhibition and activation were used to regulate the autophagy of CAF, and then their effects on PTC cell proliferation, migration and invasion were further evaluated. The in vivo effect of CAF autophagy on PTC xenograft tumor growth was evaluated.Results:CAF exhibited higher FAP expression and basal autophagy levels. PTC cells co-cultured with CAF-conditioned media showed enhanced proliferation, migration, invasion, and reduced iodine uptake. Autophagy inhibition reduced these effects, while autophagy activation further promoted them. In vivo, inhibiting CAF autophagy suppressed tumor growth.Conclusions:CAF promotes PTC cell malignancy through autophagy activation, enhancing proliferation, migration, and invasion while reducing iodine uptake.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:To explore the effects of combining APR-246 with irradiation for enhancing anti-tumor immune response against 4T1 breast cancer cells, and to develop multiple tumor treatment strategies.Methods:The control group, APR-246 group, irradiation group and irradiation combined APR-246 group were used both in the cell experiment and tumor-bearing mice experiment. The inhibitory effect of APR-246 on the proliferation of 4T1 cells was assessed by using Cell Counting Kit-8. The effect of APR-246 with irradiation on the survival rate of 4T1 cells using clone formation assay was measured. The levels of reactive oxygen species (ROS) and lipid peroxidation (LPO) in tumor cells using a 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe and a lipid peroxidation sensor, the tumor inhibition rates of different groups of tumor bearing mice were compared, and the proportions of CD4+ and CD8+ T cells and the ratio of M1/M2 macrophages were determined in the tumor microenvironment by flow cytometry.Results:Compared with irradiation group, 2, 4, 6 Gy irradiation combined APR-246 group significantly reduced the survival rates of 4T1 cells ( t = 2.89, 4.15, 2.62, P < 0.05), the 6 Gy irradiation combined APR-246 group significantly increased the levels of ROS ( t = 16.95, P < 0.05) and LPO ( t = 6.09, P < 0.05) in 4T1 cells, and significantly increased the apoptosis rate of 4T1 cells ( t = 10.99, P < 0.05). Meanwhile, from the 16 th day of tumor inoculation, the 10 Gy irradiation combined APR-246 group showed significantly inhibited tumor growth ( t = 2.38-2.91, P < 0.05) and significantly increased proportions of CD4+ and CD8+ T cells ( t = 9.96, 6.28, P < 0.05) and M1/M2 ratio ( t = 15.30, P < 0.05) in tumor tissues. Conclusions:APR-246 combined with irradiation can effectively increase ROS and LPO levels in 4T1 cells, promote tumor cell apoptosis, and induce anti-tumor immune response, thus potentially inhibiting the growth of 4T1 cells.

Objective To investigate the protective effects of secretomes released by three-dimensional cultured mesenchymal stem cells(MSCs)on neurons subjected to seawater immersion(SW)and stretch injury(SI),and to provide new insights into neuronal repair following SW combined with traumatic brain injury(TBI).Methods MSCs were cultured using the hanging drop method,and the conditioned medium(CM)containing MSCs secretomes was collected.A cellular model combining SW with SI was established using mouse hippocampal neuronal cells(HT22 cells).HT22 cells were randomly assigned to five groups:control,SI,SI+SW,SI+CM,and SI+SW+CM groups.Cell viability was assessed using the CCK-8 assay,apoptosis rate was measured by flow cytometry,cell migration ability was evaluated by scratch assay,and the expression levels of apoptosis-related proteins Bcl-2 and Bcl-2-associated protein(Bax),and ferroptosis-related proteins long-chain acyl-CoA synthetase 4(ACSL4)and cyclooxygenase-2(COX-2)were detected by Western blotting.Results Immersion in 15%seawater for 12 h significantly decreased HT22 cell viability(P<0.05).The CCK-8 assay indicated that cell viability in both the SI and SI+SW groups was significantly lower than that in control group after 12 h of treatment(P<0.05).Treatment with CM containing MSCs secretomes significantly increased cell viability in SI+CM group compared to SI group(P<0.0001),and in SI+SW+CM group compared to SI+SW group(P<0.001).Flow cytometry results revealed that the apoptosis rate in SI and SI+SW groups was significantly higher than that in control group(P<0.05 or P<0.001),while in SI+CM group was lower than that in SI group(P<0.05),and in SI+SW+CM group was lower than that in SI+SW group(P<0.05).Western blotting showed that compared to control group,SI and SI+SW groups exhibited reduced Bcl-2 expression level(P<0.01 or P<0.0001)and increased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.0001).Compared to SI group,the SI+CM group displayed increased Bcl-2 expression level(P<0.05)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.05).Compared to SI+SW group,SI+SW+CM group exhibited increased Bcl-2 expression level(P<0.01)and decreased expression levels of Bax,ACSL4,and COX-2(P<0.01 or P<0.001).Scratch assay results demonstrated that at both 12 h and 24 h,the cell migration rate in SI and SI+SW groups was significantly lower than that in control group(P<0.01 or P<0.0001),while the migration rate in SI+CM group was significantly higher than that in SI group(P<0.0001 or P<0.01),and the migration rate in SI+SW+CM group was significantly higher than that in SI+SW group(P<0.0001).Conclusion Secretomes derived from MSCs cultured using the hanging drop method can alleviate neuronal damage caused by SW and TBI,potentially offering a therapeutic approach for SW combined with TBI.