ObjectiveThis paper aims to explore the action and molecular mechanism of modified Tongluo Tangtai Formula（MTLTT） on myelin damage in diabetic peripheral neuropathy （DPN） based on network pharmacology and in vitro experiments. MethodsThe chemical components of the MTLTT were retrieved from the Traditional Chinese Medicine Systems Pharmacology （TCMSP） database and literature， and the component targets were collected from the SwissTargetPrediction database. The targets of DPN were collected from the GeneCards， OMIM， Disgenet， and GEO databases. Gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses were performed using the Metascape database， and a network diagram was constructed using Cytoscape software. The binding actions of core components with glycogen synthase kinase 3 beta （GSK-3β） and β-catenin were analyzed by Autodock Vina. An in vitro DPN model was established by high glucose-induced Schwann cells and dorsal root ganglion cells （SCs/DRGs）. The ultrastructural morphological changes of SCs and DRGs were observed by scanning electron microscope（SEM）， and the expressions of myelin-associated glycoprotein （MAG） and myelin basic protein （MBP） were detected by immunofluorescence staining. The mRNA and protein expression levels of MAG， MBP， myelin protein 0 （P0）， peripheral myelin protein 22 （PMP22）， and Wnt/β-catenin signaling pathway-related protein β-catenin， GSK-3β， Wnt family member 3α （Wnt3α）， and Wnt inhibitory factor-1 （Wif-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot. ResultsNetwork pharmacology analysis revealed that MTLTT components may treat DPN via the Wnt signaling pathway， involving key proteins such as GSK-3β， β-catenin and Wif-1. The molecular docking results indicate that atropine， apigenin， baicalein， isoflavanone， and albiflorin have good binding activity with GSK-3β， and that all 13 core components have stable binding activity with β-catenin. Cell experiments showed that compared with the blank group， SCs and DRGs in the model group exhibited severe morphological and structural abnormalities such as disintegration， shrinkage and axonal rupture， while these abnormal changes were improved after MTLTT intervention. Immunofluorescence results indicated that the fluorescence intensity of MAG and MBP was markedly decreased in the model group relative to the blank group（P<0.01）， while MTLTT treatment obviously upregulated the expression of MAG and MBP compared with the model group （P<0.01）. Real-time PCR and Western blot assays revealed that the expression levels of myelin-related molecules MAG， MBP， P0 and PMP22 were significantly reduced in the model group （P<0.05，P<0.01）， and MTLTT remarkably increased their expression levels （P<0.05）. In the Wnt/β-catenin signaling pathway， the mRNA levels of GSK-3β， Wif-1 and Wnt3α were elevated and β-catenin mRNA expression was declined in the model group （P<0.01）. Meanwhile， the protein expressions of GSK-3β and Wif-1 were upregulated， whereas those of Wnt3α and β-catenin were downregulated （P<0.01）. Compared with the model group， MTLTT at different doses reduced the mRNA and protein levels of GSK-3β and Wif-1 to varying degrees （P<0.05）， and distinctly enhanced the protein expression of Wnt3α and β-catenin（P<0.01）. ConclusionMTLTT can alleviate high glucose-induced myelin damage. Its protective mechanism may promote myelin repair by upregulating the expression of MAG， MBP， P0 and PMP22， and the therapeutic effect is possibly associated with the activation of Wnt/β-catenin signaling pathway.

ObjectiveTo investigate the impact of Buzhong Yiqitang combined with cisplatin on the proliferation of human lung adenocarcinoma （A549） cells through regulation of the pyruvate dehydrogenase kinase 1 （PDK1）/protein kinase B （Akt） signaling pathway and influence on glycolysis. MethodsTranscriptome sequencing （RNA-seq） was employed to compare the expression of glycolysis-related genes between A549 cells and cisplatin-resistant human lung adenocarcinoma cells （A549/DDP）. Small interfering RNA （siRNA） was employed to knock down PDK1， and the knockdown efficiency was verified by Western blot and Read-time PCR. The cell counting kit-8 （CCK-8） assay was used to assess the survival and viability of A549 cells under the following conditions： siRNA negative control+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）， siPDK1+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）， and siPDK1+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）+Buzhong Yiqitang （10%）-containing serum. The 20% inhibitory concentration （IC₂₀） of the siRNA negative control+cisplatin group （7.832 μmol·L^-1） was calculated and used as the subsequent cisplatin concentration. Colony formation assay was performed to evaluate the proliferation of A549 cells. Lactate and adenosine triphosphate （ATP） assay kits were used to measure lactate and ATP production. The mitochondrial membrane potential was detected with the fluorescent probe JC-1. Western blotting was conducted to examine the expression levels of PDK1， phosphorylated （p）-Akt， Akt， pyruvate kinase M2 （PKM2）， glucose transporter 1 （GLUT1）， pyruvate dehydrogenase （PDH）， and lactate dehydrogenase A （LDHA）. Confocal immunofluorescence was employed to detect PDK1 and p-Akt. ResultsRNA-seq results identified PDK1 as a highly expressed differential gene in glycolysis metabolism between A549 cells and A549/DDP cells， and it was highly expressed in tumor cells. Gene Set Enrichment Analysis （GSEA） revealed upregulated and downregulated genes in glycolysis and gluconeogenesis pathways. Western blot and RT-qPCR confirmed that PDK1-si-2 had the highest transfection efficiency， with a PDK1 knockdown rate exceeding 60%. CCK-8 assay determined the half-maximal inhibitory concentration （IC₅₀） values for each group as （30.698±5.348）， （16.372±3.562）， （13.237±1.573） μmol·L^-1， while the IC₂₀ of cisplatin in siRNA negative control-transfected A549 cells was （7.832±0.672） μmol·L^-1. Compared with the siRNA negative control group， the siRNA negative control+cisplatin group showed decreased colony formation rate， reduced lactate production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 fluorescence intensity （P<0.05）. The siPDK1 group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， PKM2， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1 group， the siPDK1+Buzhong Yiqitang group showed decreased colony formation rate， reduced lactate production， downregulated protein levels of PKM2， GLUT1， and PDK1， and reduced PDK1 fluorescence intensity （P<0.05）. The siPDK1+cisplatin group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of PKM2， GLUT1， PDK1， and PDH， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. The siPDK1+cisplatin+Buzhong Yiqitang group demonstrated decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， PKM2， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1+Buzhong Yiqitang group， the siPDK1+cisplatin group showed decreased colony formation rate， downregulated protein levels of p-Akt and PDH， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. The siPDK1+cisplatin+Buzhong Yiqitang group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， GLUT1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1+cisplatin group， the siPDK1+cisplatin+Buzhong Yiqitang group showed decreased colony formation rate， increased ATP production， and downregulated protein levels of p-Akt， PKM2， and LDHA （P<0.05）. ConclusionBuzhong Yiqitang combined with cisplatin can suppress lung adenocarcinoma cell proliferation by modulating glycolysis through the PDK1/Akt signaling pathway.

The Kirsten rat sarcoma viral oncogene homolog ( KRAS ) mutation is one of the most prevalent activating alterations in cancer. It indicates a poor overall prognosis due to its highly invasive nature. Although several KRAS inhibitors have been developed in recent years, a significant clinical challenge has emerged as a substantial proportion of patients eventually develop resistance to these therapies. Therefore, identifying determinants of drug resistance is critical for guiding treatment strategies. This review provides a comprehensive overview of the mutation landscape and molecular mechanisms of KRAS activity in various cancers. Meanwhile, it summaries the progress and prospects of small molecule KRAS inhibitors undergoing clinical trials. Furthemore, this review explores potential strategies to overcome drug resistance, with the ultimate goal of steering toward patient-centric precision oncology in the foreseeable future.
Humans ; Drug Resistance, Neoplasm/drug effects* ; Proto-Oncogene Proteins p21(ras)/metabolism* ; Mutation/genetics* ; Neoplasms/genetics* ; Antineoplastic Agents/therapeutic use*

Metastasis is a pivotal and intricate process in the progression of malignant tumors,strongly correlating with poor prognosis.Approximately 90%of cancer-related mortality is attributed to metastasis,with the five-year survival rate for patients with metastatic solid tumors ranging from 5%to 30%.Consequently,a comprehensive understanding of the underlying biological mechanisms driving metastasis is essential for unraveling its core processes and developing novel therapeutic strategies.The metastatic cascade involves tumor cells navigating numerous biological barriers,including detachment from the primary tumor,invasion of blood vessels or lymphatics,survival in circulation,extravasation into distant organs and subsequent adaptation to the microenvironment.To surmount these challenges,tumor cells undergo phenotypic changes,genetic mutations and dysregulating signaling pathways.Additionally,microenvironmental factors(such as angiogenesis,matrix remodeling and immune evasion)play a critical role,orchestrating the initiation and growth of metastatic lesions in an interdependent manner.Organ-specific metastasis,a distinct subset of metastasis,involves dynamic bidirectional interactions between tumor cells and the microenvironment of target organs.These interactions determine the selectivity of metastatic spread and drive the adaptive evolution of both the tumor and the organ,which encompasses multiple layers of cellular interactions,including cell-cell and cell-matrix signaling.Tumor cell mutations,the release of specific signaling molecules,the capacity to withstand circulatory pressures,and signaling exchanges with target organs collectively govern the selective nature of organ-specific metastasis.Furthermore,factors intrinsic to the target organ-such as its regenerative potential,metabolic profile,immune surveillance mechanisms and matrix stiffness-further facilitate the adaptive remodeling of metastatic cells within these environments.Thus,the bidirectional selection and adaptation between tumor cells and target organs form a dynamic,complex system that reshapes our understanding of metastatic tumor development.While current research emphasizes shared biological features in metastasis,the successful formation of metastatic tumors depends not only on these common mechanisms but also on the unique characteristics governing organ-specific metastasis.The interplay between generalizable and organ-specific mechanisms profoundly influences the metastatic outcome.This review aimed to consolidate our current knowledge of these shared and distinct processes,analyze the evolving understanding of the bidirectional selection between tumor cells and target organs,and assess the current status of metastatic risk prediction models for patients without metastasis.Furthermore,the paper discussed the challenges and opportunities in managing advanced-stage metastatic tumors,offering new insights and potential clinical strategies to improve prognosis and treatment outcomes.

Objective To investigate the nebulization characteristics of human interferon α1b for injection.To characterize and compare the delivery rate,total drug substance and the aerodynamic characteristics between the two types of nebulizer.Methods Connect two types of nebulizers with a breathing simulator,respectively,and simulate the breathing patterns of an infant and child.Measure the delivery rate,total delivered dose,and delivery uniformity.The aerodynamic properties of human interferon α1b for injection were evaluated by the next generation impactor(NGI).The content of human interferon α1b was quantified by double antibody sandwich ELISA.Results In the three batches of samples in infant mode,the delivery rate and total delivered dose determind by A nebulizer were 0.45,0.49,0.44 μg·min-1,3.06,3.21,3.81 μg,respectcively;and 0.12,0.14,0.16 μg·min-1,0.73,0.73,0.75 μg,respectcively by B nebulizer.In child mode,the delivery rate and total delivered doses determined by A nebulizer were 1.36,1.49,1.20 μg·min-1,7.44,7.17,and 6.54 μg,respectcively;and 0.37,0.36,0.43 μg·min-1,1.66,1.59,and 1.41 μg,respectcively by B nebulizer.In child and infant mode,the ten results of the total drug substance delivered determined by nebulizer A were both between 65%to 135%of the average.The FPD,FPF,MMAD,and GSD determined by A neublizer of three batch samples were 2.48,2.92,2.35 μg,59.0%,57.4%,59.1%,4.18,4.34,4.15 μm,1.94,1.98,2.01,respectively.The FPD,FPF,MMAD and GSD determined by B neublizer of three batches samples were 2.70,3.38,3.06 μg,67.6%,66.4%,66.3%,3.55,3.65,3.68 μm,2.03,2.04,2.06,respectively.Conclusions The data obtained in this research characterized the in vitro nebulization characteristics of human interferon α1b for injection and provided a theoretical basis and reference for in vitro study and clinical practice.The influence of different types of nebulizers on nebulization characteristics was evaluated as well.It is suggested that the quality standard of nebulizers be strictly formulated and the use of nebulizers be standardized.

Objective To investigate the nebulization characteristics of human interferon α1b for injection.To characterize and compare the delivery rate,total drug substance and the aerodynamic characteristics between the two types of nebulizer.Methods Connect two types of nebulizers with a breathing simulator,respectively,and simulate the breathing patterns of an infant and child.Measure the delivery rate,total delivered dose,and delivery uniformity.The aerodynamic properties of human interferon α1b for injection were evaluated by the next generation impactor(NGI).The content of human interferon α1b was quantified by double antibody sandwich ELISA.Results In the three batches of samples in infant mode,the delivery rate and total delivered dose determind by A nebulizer were 0.45,0.49,0.44 μg·min-1,3.06,3.21,3.81 μg,respectcively;and 0.12,0.14,0.16 μg·min-1,0.73,0.73,0.75 μg,respectcively by B nebulizer.In child mode,the delivery rate and total delivered doses determined by A nebulizer were 1.36,1.49,1.20 μg·min-1,7.44,7.17,and 6.54 μg,respectcively;and 0.37,0.36,0.43 μg·min-1,1.66,1.59,and 1.41 μg,respectcively by B nebulizer.In child and infant mode,the ten results of the total drug substance delivered determined by nebulizer A were both between 65%to 135%of the average.The FPD,FPF,MMAD,and GSD determined by A neublizer of three batch samples were 2.48,2.92,2.35 μg,59.0%,57.4%,59.1%,4.18,4.34,4.15 μm,1.94,1.98,2.01,respectively.The FPD,FPF,MMAD and GSD determined by B neublizer of three batches samples were 2.70,3.38,3.06 μg,67.6%,66.4%,66.3%,3.55,3.65,3.68 μm,2.03,2.04,2.06,respectively.Conclusions The data obtained in this research characterized the in vitro nebulization characteristics of human interferon α1b for injection and provided a theoretical basis and reference for in vitro study and clinical practice.The influence of different types of nebulizers on nebulization characteristics was evaluated as well.It is suggested that the quality standard of nebulizers be strictly formulated and the use of nebulizers be standardized.

Metastasis is a pivotal and intricate process in the progression of malignant tumors,strongly correlating with poor prognosis.Approximately 90%of cancer-related mortality is attributed to metastasis,with the five-year survival rate for patients with metastatic solid tumors ranging from 5%to 30%.Consequently,a comprehensive understanding of the underlying biological mechanisms driving metastasis is essential for unraveling its core processes and developing novel therapeutic strategies.The metastatic cascade involves tumor cells navigating numerous biological barriers,including detachment from the primary tumor,invasion of blood vessels or lymphatics,survival in circulation,extravasation into distant organs and subsequent adaptation to the microenvironment.To surmount these challenges,tumor cells undergo phenotypic changes,genetic mutations and dysregulating signaling pathways.Additionally,microenvironmental factors(such as angiogenesis,matrix remodeling and immune evasion)play a critical role,orchestrating the initiation and growth of metastatic lesions in an interdependent manner.Organ-specific metastasis,a distinct subset of metastasis,involves dynamic bidirectional interactions between tumor cells and the microenvironment of target organs.These interactions determine the selectivity of metastatic spread and drive the adaptive evolution of both the tumor and the organ,which encompasses multiple layers of cellular interactions,including cell-cell and cell-matrix signaling.Tumor cell mutations,the release of specific signaling molecules,the capacity to withstand circulatory pressures,and signaling exchanges with target organs collectively govern the selective nature of organ-specific metastasis.Furthermore,factors intrinsic to the target organ-such as its regenerative potential,metabolic profile,immune surveillance mechanisms and matrix stiffness-further facilitate the adaptive remodeling of metastatic cells within these environments.Thus,the bidirectional selection and adaptation between tumor cells and target organs form a dynamic,complex system that reshapes our understanding of metastatic tumor development.While current research emphasizes shared biological features in metastasis,the successful formation of metastatic tumors depends not only on these common mechanisms but also on the unique characteristics governing organ-specific metastasis.The interplay between generalizable and organ-specific mechanisms profoundly influences the metastatic outcome.This review aimed to consolidate our current knowledge of these shared and distinct processes,analyze the evolving understanding of the bidirectional selection between tumor cells and target organs,and assess the current status of metastatic risk prediction models for patients without metastasis.Furthermore,the paper discussed the challenges and opportunities in managing advanced-stage metastatic tumors,offering new insights and potential clinical strategies to improve prognosis and treatment outcomes.

Objective:To investigate the dosimetric feasibility of cardiac stereotactic body radiotherapy (CSBRT) using volumetric-modulated arc radiotherapy (VMAT)-like technique on the Unity MR-linac.Methods:A retrospective analysis was conducted on 12 refractory arrhythmia patients who underwent CSBRT at West China Hospital, Sichuan University, from April 2021 to December 2022. Four different treatment plans were designed for each patient based on the average phase of 4DCT: VMAT plan based on a linear accelerator, VMAT-like plan with magnetic field based on Unity, VMAT-like plan without magnetic field based on a Unity, and static intensity-modulated radiation therapy (IMRT) plan with 7 beams based on Unity. The VMAT-like plans used static IMRT beams evenly distributed at 30 angles. Evaluation metrics included conformity index (CI), homogeneity index (HI), gradient index (GI), dosimetric parameters of organs at risk (OAR), optimization time, and monitor unit (MU). These metrics among groups were analyzed using the Wilcoxon paired signed-rank test.Results:The CI of the VMAT plan was better than that of the VMAT-like plan with magnetic field (0.84 vs. 0.81, P=0.005). The GI of the VMAT-like plan without magnetic field was significantly lower than that of VMAT-like plan with magnetic field ( P=0.015). The CI and HI of the IMRT plan were both inferior to VMAT-like plan with magnetic field ( P=0.034). The average dose to the heart and lungs in VMAT-like plan with magnetic field was higher than those in the VMAT plan and VMAT-like plan without magnetic field (both P<0.05). The magnetic field significantly increased the dose to the skin, tissue-lung interface, and outside the margins (all P<0.05). The optimization time of the VMAT-like plan with magnetic field was longer than that of the IMRT plan, but it required fewer MUs ( P=0.001). The quality of the IMRT plans was poor and did not meet clinical requirements. Conclusion:The quality of CSBRT plans based on VMAT-like techniques meets clinical requirements, but attention must be paid to the dosimetric effects of the magnetic field.

Objective:To investigate the screening values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus (HR-HPV) testing for high-grade cervical lesions. Methods:The clinical data of 622 patients who underwent cervical thin-layer liquid-based cytology (TCT) and HR-HPV testing in General Hospital of Taiyuan Iron and Steel (Group) Co., Ltd. from March 2019 to July 2021 were retrospectively analyzed. The remaining cytological specimens were detected by P16/Ki-67 double staining and P16 INK4α single staining. Among them, 334 patients with TCT results suggesting atypical squamous cell of undetermined significance (ASCUS) and above and HPV-positive underwent colposcopy pathological biopsy. Using pathological results as reference, the positive predictive value, sensitivity, specificity and accuracy of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of high-grade squamous intraepithelial neoplasia (HSIL) and cervical cancer were compared. Results:Taking the results of histopathology as references, combined with the results of TCT, 31 of 622 patients were HSIL, of which 22 (71.0%) were positive for P16/Ki-67 double staining, 23 (74.2%) were positive for P16 INK4α single staining, and 25 (80.6%) were positive for HR-HPV testing; 4 cases were cervical cancer, and the positive rates of the three detection methods were all 100.0% (4/4). Among 622 patients, the positive rates of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of HSIL and cervical cancer were 13.99% (87/622), 25.40% (158/622) and 21.38% (133/622); the positive predictive values were 29.89%, 17.09% and 21.08%; the accuracies were 91.19%, 78.94% and 83.28%; the specificities were 89.77%, 77.98% and 82.46%; the sensitivities were 74.29%, 77.14% and 82.86%. The positive rate, positive predictive value, specificity and accuracy of P16/Ki-67 double staining were higher than those of P16 INK4α single staining and HR-HPV testing, and the differences were statistically significant ( z values were -5.062 and -3.418, 2.328 and 2.450, 5.436 and 3.570, 6.043 and 4.161, all P < 0.05); the sensitivity of HR-HPV testing was higher than that of P16/Ki-67 double staining and P16 INK4α single staining, but the differences were not statistically significant ( z values were -0.890 and 1.017, both P > 0.05). Conclusions:HR-HPV testing is more suitable for primary cervical lesion screening; P16/Ki-67 double staining can be used as a potential combined cell screening tool or an effective triage tool; P16 INK4α single staining has certain limitations.

Objective:To retrospectively analyze the setup errors of thermoplastic head and shoulder molds alone or combined with vacuum pad in hypofractionated stereotactic radiotherapy (HFSRT) for non-small cell lung cancer (NSCLC) with brain metastases.Methods:Fifty-four NSCLC patients with brain metastases who received HFSRT from 2017 to 2019 were enrolled in this study. Twenty-four patients were fixed with thermoplastic head and shoulder molds (group A), and 30 patients were fixed with thermoplastic head and shoulder molds plus vacuum pad (group B). The interfraction and intrafraction setup errors were acquired from cone-beam CT online image registration before and after the HFSRT. Optical surface system was applied in monitoring the intrafraction setup errors. The setup errors in each direction between two groups were analyzed by independent samples t-test. Results:For the interfraction setup errors of the whole group, the proportion of the horizontal setup errors of ≥3mm was 7.0% to 15.4% and 7.0% to 12.6% for the rotation setup errors of ≥2°. In group A, the anteroposterior setup error was (1.035±1.180)mm, significantly less than (1.512±0.955)mm in group B ( P=0.009). In group A, the sagittal rotation setup error was 0.665°±0.582°, significantly less than 0.921°±0.682° in group B ( P=0.021). For the intrafraction setup errors of the whole group, the proportion of horizontal setup errors of ≥1mm was 0% to 0.7%, whereas no rotation setup error of ≥1° were observed. In group B, bilateral, anteroposterior and sagittal rotation setup errors were (0.047±0.212)mm, (0.023±0.152)mm and 0.091°±0.090°, significantly less compared with (0.246±0.474)mm, (0.140±0.350)mm and 0.181°±0.210° in group A ( P=0.004, P=0.020, P=0.001), respectively. Optical surface monitoring data were consistent with the obtained results. Conclusions:Thermoplastic head and shoulder molds (with or without vacuum pad) combined with online image registration and six-dimensional robotic couch correction can be applied in HFSRT for brain metastases from NSCLC. The intrafraction setup errors in group B are smaller than those in group A. Optical surface system has certain value in monitoring the intrafractional movement.