Triple-negative breast cancer (TNBC) represents a distinctive subtype, characterized by the absence of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor 2 (HER2). Due to its high inter-tumor and intra-tumor heterogeneity, TNBC poses significant chanllenges for personalized diagnosis and treatment. The advant of clustered regular interspaced short palindromic repeats (CRISPR) technology has profoundly enhanced our understanding of the structure and function of the TNBC genome, providing a powerful tool for investigating the occurrence and development of diseases. This review focuses on the application of CRISPR/Cas technology in the personalized diagnosis and treatment of TNBC. We begin by discussing the unique attributes of TNBC and the limitations of current diagnostic and treatment approaches: conventional diagnostic methods provide limited insights into TNBC, while traditional chemotherapy drugs are often associated with low efficacy and severe side effects. The CRISPR/Cas system, which activates Cas enzymes through complementary guide RNAs (gRNAs) to selectively degrade specific nucleic acids, has emerged as a robust tool for TNBC research. This technology enables precise gene editing, allowing for a deeper understanding of TNBC heterogeneity by marking and tracking diverse cell clones. Additionally, CRISPR facilitates high-throughput screening to promptly identify genes involved in TNBC growth, metastasis, and drug resistance, thus revealing new therapeutic targets and strategies. In TNBC diagnostics, CRISPR/Cas was applied to develop molecular diagnostic systems based on Cas9, Cas12, and Cas13, each employing distinct detection principles. These systems can sensitively and specifically detect a variety of TNBC biomarkers, including cell-specific DNA/RNA and circulating tumor DNA (ctDNA). In the realm of precision therapy, CRISPR/Cas has been utilized to identify key genes implicated in TNBC progression and treatment resistance. CRISPR-based screening has uncovered potential therapeutic targets, while its gene-editing capabilities have facilitated the development of combination therapies with traditional chemotherapy drugs, enhancing their efficacy. Despite its promise, the clinical translation of CRISPR/Cas technology remains in its early stages. Several clinical trials are underway to assess its safety and efficacy in the treatment of various genetic diseases and cancers. Challenges such as off-target effects, editing efficiency, and delivery methods remain to be addressed. The integration of CRISPR/Cas with other technologies, such as 3D cell culture systems, human induced pluripotent stem cells (hiPSCs), and artificial intelligence (AI), is expected to further advance precision medicine for TNBC. These technological convergences can offer deeper insights into disease mechanisms and facilitate the development of personalized treatment strategies. In conclusion, the CRISPR/Cas system holds immense potential in the precise diagnosis and treatment of TNBC. As the technology progresses and becomes more costs-effective, its clinical relevance will grow, and the translation of CRISPR/Cas system data into clinical applications will pave the way for optimal diagnosis and treatment strategies for TNBC patients. However, technical hurdles and ethical considerations require ongoing research and regulation to ensure safety and efficacy.

Lung cancer is the most common malignant tumor worldwide, ranking first in both incidence and mortality rates. According to the latest statistics from the International Agency for Research on Cancer (IARC), approximately 2.5 million new cases and around 1.8 million deaths from lung cancer occurred in 2022, placing a tremendous burden on global healthcare systems. The high mortality rate of lung cancer is closely linked to its subtle early symptoms, which often lead to diagnosis at advanced stages. This not only complicates treatment but also results in substantial economic losses. Current treatment options for lung cancer include surgery, radiotherapy, chemotherapy, targeted drug therapy, and immunotherapy. Among these, immunotherapy has emerged as the most groundbreaking advancement in recent years, owing to its unique antitumor mechanisms and impressive clinical benefits. Unlike traditional therapies such as radiotherapy and chemotherapy, immunotherapy activates or enhances the patient’s immune system to recognize and eliminate tumor cells. It offers advantages such as more durable therapeutic effects and relatively fewer toxic side effects. The main approaches to lung cancer immunotherapy include immune checkpoint inhibitors, tumor-specific antigen-targeted therapies, adoptive cell therapies, cancer vaccines, and oncolytic virus therapies. Among these, immune checkpoint inhibitors and tumor-specific antigen-targeted therapies have received approval from the U.S. Food and Drug Administration (FDA) for clinical use in lung cancer, significantly improving outcomes for patients with advanced non-small cell lung cancer. Although other immunotherapy strategies are still in clinical trials, they show great potential in improving treatment precision and efficacy. This article systematically reviews the latest research progress in lung cancer immunotherapy, including the development of novel immune checkpoint molecules, optimization of treatment strategies, identification of predictive biomarkers, and findings from recent clinical trials. It also discusses the current challenges in the field and outlines future directions, such as the development of next-generation immunotherapeutic agents, exploration of more effective combination regimens, and the establishment of precise efficacy prediction systems. The aim is to provide a valuable reference for the continued advancement of lung cancer immunotherapy.

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline, functional impairment, and neuropsychiatric symptoms. It represents the most prevalent form of dementia among the elderly population. Accumulating evidence indicates that oxidative stress plays a pivotal role in the pathogenesis of AD. Notably, elevated levels of oxidative stress have been observed in the brains of AD patients, where excessive reactive oxygen species (ROS) can cause extensive damage to lipids, proteins, and DNA, ultimately compromising neuronal structure and function. Amyloid β‑protein (Aβ) has been shown to induce mitochondrial dysfunction and calcium overload, thereby promoting the generation of ROS. This, in turn, exacerbates Aβ aggregation and enhances tau phosphorylation, leading to the formation of two pathological features of AD: extracellular Aβ plaque deposition and intracellular neurofibrillary tangles (NFTs). These events ultimately culminate in neuronal death, forming a vicious cycle. The interplay between oxidative stress and these pathological processes constitutes a core link in the pathogenesis of AD. The signaling pathways mediating oxidative stress in AD include Nrf2, RCAN1, PP2A, CREB, Notch1, NF‑κB, ApoE, and ferroptosis. Nrf2 signaling pathway serves as a key regulator of cellular redox homeostasis, exerts important antioxidant capacity and protective effects in AD. RCAN1 signaling pathway, as a calcineurin inhibitor, and modulates AD progression through multiple mechanisms. PP2A signaling pathway is involved in regulating tau phosphorylation and neuroinflammation processes. CREB signaling pathway contributes to neuroplasticity and memory formation; activation of CREB improves cognitive function and reduce oxidative stress. Notch1 signaling pathway regulates neuronal development and memory, participates in modulation of Aβ production, and interacts with Nrf2 toco-regulate antioxidant activity. NF‑κB signaling pathway governs immune and inflammatory responses; sustained activation of this pathway forms “inflammatory memory”, thereby exacerbating AD pathology. ApoE signaling pathway is associated with lipid metabolism; among its isoforms, ApoE-ε4 significantly increases the risk of AD, leading to elevated oxidative stress, abnormal lipid metabolism, and neuroinflammation. The ferroptosis signaling pathway is driven by iron-dependent lipid peroxidation, and the subsequent release of lipid peroxidation products and ROS exacerbate oxidative stress and neuronal damage. These interconnected pathways form a complex regulatory network that regulates the progression of AD through oxidative stress and related pathological cascades. In terms of therapeutic strategies targeting oxidative stress, among the drugs currently used in clinical practice for AD treatment, memantine and donepezil demonstrate significant therapeutic efficacy and can improve the level of oxidative stress in AD patients. Some compounds with antioxidant effects (such asα-lipoic acid and melatonin) have shown certain potential in AD treatment research and can be used as dietary supplements to ameliorate AD symptoms. In addition, non-drug interventions such as calorie restriction and exercise have been proven to exerted neuroprotective effects and have a positive effect on the treatment of AD. By comprehensively utilizing the therapeutic characteristics of different signaling pathways, it is expected that more comprehensive multi-target combination therapy regimens and combined nanomolecular delivery systems will be developed in the future to bypass the blood-brain barrier, providing more effective therapeutic strategies for AD.

ObjectiveIn nature, objects cast shadows due to illumination, forming the basis for stereoscopic perception. Birds need to adapt to changes in lighting (meaning they can recognize stereoscopic shapes even when shadows look different) to accurately perceive different three-dimensional forms. However, how neurons in the key visual brain area in birds handle these lighting changes remains largely unreported. In this study, pigeons (Columba livia) were used as subjects to investigate how neurons in pigeon’s ventrolateral mesopallium (MVL) represent stereoscopic shapes consistently, regardless of changes in lighting. MethodsVisual cognitive training combined with neuronal recording was employed. Pigeons were first trained to discriminate different stereoscopic shapes (concave/convex). We then tested whether and how light luminance angle and surface appearance of the stereoscopic shapes affect their recognition accuracy, and further verify whether the results rely on specify luminance color. Simultaneously, neuronal firing activity of neurons was recorded with multiple electrode array implanted from the MVL during the presentation of difference shapes. The response was finally analyzed how selectively they responded to different stereoscopic shapes and whether their selectivity was affected by the changes of luminance condition (like lighting angle) or surface look. Support vector machine (SVM) models were trained on neuronal population responses recorded under one condition (light luminance angle of 45°) and used to decode responses under other conditions (light luminance angle of 135°, 225°, 315°) to verify the invariance of responses to different luminance conditions. ResultsBehavioral results from 6 pigeons consistently showed that the pigeons could reliably identify the core 3D shape (over 80% accuracy), and this ability wasn’t affected by changes in light angle or surface appearance. Statistical analysis of 88 recorded neurons from 6 pigeons revealed that 83% (73/88) showed strong selectivity for specific 3D shapes (selectivity index>0.3), and responses to convex shapes were consistently stronger than to concave shapes. These shape-selective responses remained stable across changes in light angle and surface appearance. Neural patterns were consistent under both blue and orange lighting. The decoding accuracy achieves above 70%, suggesting stable responses under different conditions (e.g., different lighting angles or surface appearance). ConclusionNeurons in the pigeon MVL maintain a consistent neural encoding pattern for different stereoscopic shapes, unaffected by illumination or surface appearance. This ensures stable object recognition by pigeons in changing visual environments. Our findings provide new physiological evidence for understanding how birds achieve stable perception (“invariant neural representations”) while coping with variations in the visual field.

Objective To analyze the clinical characteristics of recipients with interstitial lung disease (ILD) who underwent second lung transplantation and summarize the diagnostic and therapeutic experience. Methods A retrospective analysis was conducted on the clinical data of 14 patients who underwent first and second lung transplants for ILD at the First Affiliated Hospital of Guangzhou Medical University from January 2015 to December 2024. The preoperative conditions, intraoperative events, postoperative treatments and prognoses of the first and second lung transplantation were compared, and the postoperative survival of ILD patients after the second lung transplantation was analyzed. Results Among the 14 recipients of the second lung transplant, 13 underwent the procedure due to chronic lung allograft dysfunction, and 1 due to airway complications. The median interval time from the first to the second lung transplant was 32 (2, 80) months. Before the second transplantation, 2 recipients required endotracheal intubation and mechanical ventilation, and 2 required endotracheal intubation, mechanical ventilation, and extracorporeal membrane oxygenation (ECMO) support. The surgical time for the second lung transplantation was longer than that for the first, with increased intraoperative red blood cell and plasma transfusion volumes, the proportion of ECMO support during the second lung transplantation was higher than that during the first (all P<0.05). However, the cold ischemia time for one-sided lung transplant completion in the first lung transplant was similar to that in the second lung transplantation (P>0.05). The median follow-up time after the second lung transplantation was 32 (1, 63) months. The 1-month, 6-month and 1-year survival rates after the second lung transplantation were 79%, 57% and 50%, respectively, with causes of death being infection, multiple organ failure and gastrointestinal bleeding. Conclusions For ILD patients undergoing second lung transplantation after the first lung transplantation, the second lung transplantation is more challenging, with longer surgical time and higher intraoperative blood loss. It requires higher surgical skills and perioperative management. Non-emergency second transplantation may still achieve good results.

Vascular calcification is a highly regulated process of ectopic calcification in cardiovascular system while no effective intervention can be clinically performed up to date.As vascular calcification undergoes a common regulatory mechanism within bone formation,bone morphogenetic protein 7(BMP-7)main-tains contractile phenotype of vascular smooth muscle cells and further inhibits vascular calcification via promoting the process of osteoblast differentiation,reducing ectopic calcification pressure by increasing bone formation and reducing bone resorption.This work systematically reviews the role of BMP-7 in vascular calcifi-cation and the possible mechanism,and their current clinical application as well.The current proceedings may help develope early diagnostic strategy and therapeutic treatment with BMP-7 as a new molecular marker and potential drug target.The expec-tation could achieve early prevention and intervention of vascular calcification and improve poor prognosis on patients.

Objective:To investigate the expression level of long non-coding RNA (lncRNA) CTC-338M12.4 in tongue squamous cell carcinoma tissues, and its effects on the cell cycle, cell proliferation, and migration of tongue squamous cell carcinoma cells in vitro as well as its molecular mechanisms.Methods:The Gene Expression Omnibus (GEO) database was used to obtain the lncRNA series data set GSE139869, and the differential expression of CTC-338M12.4 in tongue squamous cell carcinoma tissues and adjacent tissues was analyzed. The transcriptional expression levels of CTC-338M12.4 in human immortalized oral keratinocytes (HOK) and tongue squamous cell carcinoma cell lines HN13, TSCCa, CAL-27, Tca8113, SCC15 were detected by using reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR). CAL-27 cells with the lowest expression level of CTC-338M12.4 were selected and were divided into the control group (co-transfected with vectors containing negative sequence) and CTC-338M12.4 group (co-transfected with CTC-338M12.4 overexpression vectors). The proliferation ability of CAL-27 cells in each group was detected by using cell colony formation assay, and the cell cycle distribution of CAL-27 cells was detected by using flow cytometry. The migration ability of CAL-27 cells was detected by using scratch test. The lncACTdb database was used to predict the complementary binding sites between CTC-338M12.4 and miRNA-27a-5p (miR-27a-5p), and dual-luciferase reporter gene assay was used to verify. The expression level of miR-27a-5p in CAL-27 cells of all groups was detected by using qRT-PCR. The protein expression level of related factors on JAK/STAT signaling pathway in CAL-27 cells of all groups was detected by using Western blot.Results:Analysis of GEO database data showed that transcriptional level CTC-338M12.4 in tongue squamous cell carcinoma tissues was lower than that in adjacent tissues, and the difference was statistically significant ( P < 0.01). Transcriptional level CTC-338M12.4 in tongue squamous cell carcinoma HNl3, TSCCa, CAL-27, Tca8113, and SCC15 cells was lower than that in HOK cells, and the differences were statistically significant (all P < 0.05). The transcriptional level relative expression level of CTC-338M12.4 in CAL-27 cells in the CTC-338M12.4 group was higher than that in the control group, and the difference was statistically significant ( P < 0.01). Cell colony formation assay showed that the colong number of CAL-27 cell in the CTC-338M12.4 group was less than that in the control group [(51±10) vs. (114±21)], and the difference was statistically significant ( t = 2.71, P = 0.035). Flow cytometry showed that the proportion of G 0/G 1 phase cells in CAL-27 cells in the CTC-338M12.4 group was higher than that in the control group [(64±3)% vs. (43±4)%], and the difference was statistically significant ( t = 4.87, P = 0.003). The scratch test showed that when scratching, the scratch width of both groups was similar ( P > 0.05); after scratch for 25 h, the scratch width of CAL-27 cells in the CTC-338M12.4 group was wider than that in the control group [(133±15) μm vs. (64±10) μm], and the difference was statistically significant ( t = 3.78, P = 0.009). The dual luciferase reporter gene assay showed that the relative luciferase activity of CAL-27 cells co-transfected with wild-type CTC-338M12.4 sequence and miR-27a-5p sequence was lower than that of CAL-27 cells co-transfected with wild-type CTC-338M12.4 sequence and miR-27a-5p irrelevant sequence, and the difference was statistically significant ( P < 0.001). The relative expression level of transcriptional level miR-27a-5p of CAL-27 cells in the CTC-338M12.4 group was lower than that in the control group, and the difference was statistically significant ( P = 0.003). Western blot showed that the protein expression levels of JAK/STAT signaling pathway p-JAK, p-STAT, p-Raf, p-ERK, and p-mTOR were lower than those in the control group. Conclusions:The level of lncRNA CTC-338M12.4 is low in tongue squamous cell carcinoma. CTC-338M12.4 mediates the inactivation of JAK/STAT signaling pathway via inhibiting miR-27a-5p expression in tongue squamous cell carcinoma CAL-27 cells, thereby leading to cell cycle arrest and inhibiting the cell proliferation and migration of CAL-27 cells.

Objective To clarify the understanding of types of laboratories and manufacturers for GB 9706.1-2020 Medical electrical equipment-Part 1:General requirements for basic safety and essential performance by laboratory proficiency testing for creepage distance and electrical clearance test.Methods An operation guide was formed according to the testing program in GB 9706.1-2020,and the homogeneity and stability of the samples were evaluated according to CNAS-GL003:2018 Guidance on Evaluating the Homogenneity and Stability of Samples Used for Proficiency Testing.Robust statistic methods were used to assess the quantitative parameters of the test results of the participating laboratories according to the requirements in GB/T 28043-2019 Statistical methods for use in proficiency testing by interlaboratory comparison;the results reported by the expert laboratories were used as the specified values of the qualitative parameters.SPSS 25.0 statistical software was used for data analysis.Results All the results of the crreepage distance and electrical clearance tests met the requirements for homogeneity and stability.Of the 46 laboratories involved in,37 ones did have comprehensive satisfactory determinations while the remained 9 ones not.Conclusion Some laboratories don't behave well in understanding the standard,which have to be reformed accordingly to enhance their proficiencies.[Chinese Medical Equipment Journal,2024,45(10):54-59]

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the biomechanical stabilities of different internal screw fixations for Sanders type ⅡB calcaneal fractures using finite element analysis.Methods:Based on the CT scan data of a male volunteer's calcaneus (26 years old, 173 cm tall, weighing 70 kg), a three-dimensional finite element model of the calcaneus was established using digital medical software such as Mimics 22.0, Geomagic 12.0, Solidworks 2019, and ANSYS Workbench 2022. A three-dimensional finite element model of Sanders type ⅡB calcaneal fracture was also established. The following 3 kinds of screws were used: ① longitudinal screws: from the calcaneal tuberosity to the anterior part; ② transverse screws: from the lateral wall of the calcaneus to the sustentaculum tali; ③ vertical screws: from the base of the calcaneus to the talar dome, perpendicular to the posterior articular surface. The scheme of screw configuration was as follows: the longitudinal screws were divided into the upper, middle, and lower groups based on their entry points from the upper, middle, and lower parts of the calcaneal tuberosity. Based on different configurations (double transverse screws + single vertical screw, single transverse screw + single vertical screw, double transverse screws without a vertical screw, and single transverse screw without a vertical screw) 4 subgroups were further subdivided, resulting in a total of 12 models (a, b, c, d, e, f, g, h, i, j, k, l). Forces of 420 N and 200 N were applied to the posterior and middle subtalar joints, respectively, to simulate the load on the calcaneus during single-leg standing in adults. In each model, total deformation displacement, relative displacement of the sustentaculum tali fracture gap, relative displacement of the anterior fracture gap, relative displacement of the articular surface fracture gap, peak stress at the fracture ends and peak screw stress were analyzed.Results:The total deformation displacement was the lowest in models i and j, while models k and l exhibited the highest deformation displacement. The maximum deformation values in the remaining models varied only slightly. The average relative displacement of the sustentaculum tali fracture gap was the lowest in the upper group (0.27 mm) and relatively higher in the lower group, with the greatest variation seen in models k and l. The average relative displacement of the anterior fracture gap was the smallest in the lower group (0.16 mm), with models i and k performing the best. The relative displacement of the articular surface fracture gap remained at a low level across all models. The upper group had the lowest average peak stress at the fracture ends (60.20 MPa), while the middle and lower groups had similar values (103.88 MPa and 99.76 MPa, respectively). The upper group had the lowest average screw stress peak value (222.77 MPa), whereas models k and l in the lower group had the highest stress peaks, reaching 331.48 MPa and 349.82 MPa, respectively.Conclusions:Model i (inferior longitudinal screw + double transverse screws + single vertical screw) can be recommended in clinic because it demonstrates balanced performance in displacement and stress and a notable advantage in reducing overall deformation displacement and anterior fracture gap displacement. As longitudinal, vertical, and transverse screws each play their distinct role in calcaneal fracture fixation, surgeons should select an appropriate screw configuration based on the patient's needs.