OBJECTIVES: To evaluate the inhibitory effect of oridonin against proliferation of pancreatic cancer cells and the mechanism underlying the synergistic effect of low-intensity pulsed ultrasound (LIPUS). METHODS: PANC-1 cells treated with different concentrations of oridonin were examined for changes in cell proliferation using CCK-8 assay and in MDA, GSH and ATP levels using flow cytometry. The protein expressions of GPX4, Nrf2 and HO-1 in the treated cells were detected with Western blotting. The effect of Fer-1, a ferroptosis inhibitor, on proliferation of oridonin-treated cells were assessed, and the effects of oridonin combined with LIPUS on PIEZO1 protein expression was evalauted using Western blotting. A C57BL/6J mouse model bearing pancreatic cancer cell xenograft was established and treated with oridonin, LIPUS, or both, and the histological changes in the tumor tissues and tumor cell proliferation were examined with HE staining and immunohistochemistry for Ki67; the changes in GPX4 expression in the tumor tissues were detected using Western blotting and immunofluorescence staining. RESULTS: In PANC-1 cells, oridonin treatment significantly inhibited cell proliferation, increased intracellular Fe²⁺, ROS, and MDA levels, and decreased GSH and ATP levels. Oridonin also resulted in lowered GPX4 and increased HO-1 and Nrf2 protein expression levels in the cells. The combined treatment with LIPUS signficiantly enhanced the inhibitory effect of oridonin on PANC-1 cell viability in vitro and on xenograft growth in the mouse models, resulting also in more obvious reduction of the intensity of Ki67 staining and GPX4 protein expression and more pronounced increase of PIEZO1 protein expression in the tumor tissues in the mouse models. CONCLUSIONS LIPUS enhances the effect of oridonin to promote ferroptosis of pancreatic cancer cells by activating PIEZO1 through the Nrf2/HO-1/GPX4 pathway.
Ferroptosis/drug effects* ; Animals ; Pancreatic Neoplasms/metabolism* ; NF-E2-Related Factor 2/metabolism* ; Humans ; Cell Line, Tumor ; Mice ; Heme Oxygenase-1/metabolism* ; Diterpenes, Kaurane/pharmacology* ; Cell Proliferation/drug effects* ; Mice, Inbred C57BL ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Ion Channels/metabolism* ; Ultrasonic Waves ; Signal Transduction

Objective:Vascular cell adhesion molecule 1(VCAM1)is involved in a series of physiological and pathological processes,such as immune and inflammatory response,tumor cell metastasis and invasion.But under trau-matic brain injury(TBI),specific types of cells with VC AM1 expression and the related functions are not clear.In order to further explore the specific functions of VCAM1 involved in TBI,this study constructed reporter mice of VCAM1 to explore the response of VCAM1 to TBI in detail.Methods:VCAM1-Cre/ERT2::Ai14 reporter mice were constructed by gene targeting technology and Cre/loxP system,the labeled cell types and labeling efficiency were validated by im-munofluorescence staining and reporter mice hybridization.The stab wound model was used to simulate TBI to induce the changes of VCAM1 expression in cells,and the characteristics of VCAM1 positive astrocytes were detected by immu-nofluorescence staining and fluorescent probe labeling.Results:The labeling efficiency of VCAM1-Cre/ERT2::Ai14 re-porter mice was higher than that of VCAM1 antibody as was seen by labeling of more endothelial cells of blood vessels and unique astrocytes.The distribution of these astrocytes was specific,for example in the nucleus accumbens,amygda-la,hypothalamus,and paraventricular fiber systems.TBI could significantly induce the expression of VCAM1 in astro-cytes(P＜0.0001).These induced astrocytes developed reactive qualities,including somal hypertrophy,GFAP ex-pression and proliferative ability.Conclusion:VCAM1-Cre/ERT2::Ai14 reporter mice could label cells with VCAM1 expression more sensitively,so they were more effective tools for observing expression and function of VCAM1.The up-regulation of VCAM1 expression in astrocytes after TBI surgery suggested that VCAM1 was an inflammatory response molecule in astrocytes,we recommended it as a new molecular indicator of reactive astrocytes.

Depression is closely associated with a neuro-endocrine-immune(NEI)system imbalance.The"excessive vitality leading to restraint and counter-regulation(Kang Hai Cheng Zhi)"theory elucidates the self-regulating mechanism for maintaining dynamic equilibrium in the body,and serves as an importance principle guiding treatment formulation and medication selection.Based on the correlation between NEI system imbalance and the traditional Chinese medicine pathogenesis of depression,and integrating the"Kang Hai Cheng Zhi"theory,the author posits that the pathogenesis of depression lies in overactive liver invading spleen,earth dampness impeding wood′s ascendancy,and disharmony between body and mind,as well as imbalance in storage and discharge functions of liver and kidney,disharmony between Yin and Yang,and disrupted counter-regulation.This dosely aligns with two key pathological methanisms at the micro level:microglial-limbic system homeostatic imbalance and hypothalamic-pituitary-adrenal axis-inflammatory circuit dysregulation.Clinically,the treatment principle for depression adheres to supporting the counter-regulation to restrain excess,with herbal interventions using strategies such as restraining wood to support earth,dredging earth to unblock wood,and harmonizing pivotal functions,as well as nourishing water to nurture wood,warming kidney to tonify liver,and relieving depression to calm the spirit.These approaches aim to regulate the liver,spleen,and kidney,embodying the core therapeutic tenet of"striving for equilibrium,"thereby restoring the body′s self-regulating capability.

Objective:To explore the effects of deep hyperthermia on chemotherapy-related adverse effects and immune-inflammatory indicators in the patients undergoing postoperative adjuvant chemotherapy for colorectal cancer.Methods:This retrospective study included 52 patients who underwent surgery for colorectal cancer at the Affiliated Zhongshan Hospital of Dalian University from September 2021 to December 2023. The patients were divided into two groups based on treatment method: the combination group ( n=29) received postoperative adjuvant chemotherapy combined with deep hyperthermia, while the chemotherapy group ( n=23) received postoperative adjuvant chemotherapy alone. Both groups were treated with the XELOX regimen (oxaliplatin + capecitabine). The degree of bone marrow suppression during treatment was assessed by analyzing peripheral blood parameters, including hemoglobin, leukocyte count, neutrophil count, and platelet count. Immune-inflammatory indicators, including complement, procalcitonin (PCT), interleukin-6 (IL-6), systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR), were compared before and after treatment in both groups to evaluate the effects of deep hyperthermia on the immune-inflammatory response. Chi-square test or Fisher's exact test (two-tailed) was used to compare bone marrow suppression rates, and the immune-inflammatory indicators between the two groups were compared using t-tests or non-parametric tests, depending on whether the data conformed to a normal distribution. Results:In terms of myelosuppression, the incidence rates of moderate to severe decreases in leukocytes, neutrophils, platelets, and hemoglobin in the combination group were 31%, 31%, 21%, and 14%, respectively, compared to 52%, 61%, 48%, and 9% in the chemotherapy group. The change in PCT levels before and after treatment was significantly greater in the combination group than in the chemotherapy group ( P = 0.010). Both the combination group and the chemotherapy group showed significant reductions in SII, NLR and PLR after treatment, and the differences were statistically significant (all P < 0.05). The change in NLR before and after treatment was significantly greater in the combination group than in the chemotherapy group ( P = 0.031). Conclusions:Deep hyperthermia can alleviate chemotherapy-induced adverse effects such as thrombocytopenia and neutropenia in patients undergoing postoperative adjuvant chemotherapy for colorectal cancer. It also appears to improve the inflammatory response in these patients.

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.

Depression is closely associated with a neuro-endocrine-immune(NEI)system imbalance.The"excessive vitality leading to restraint and counter-regulation(Kang Hai Cheng Zhi)"theory elucidates the self-regulating mechanism for maintaining dynamic equilibrium in the body,and serves as an importance principle guiding treatment formulation and medication selection.Based on the correlation between NEI system imbalance and the traditional Chinese medicine pathogenesis of depression,and integrating the"Kang Hai Cheng Zhi"theory,the author posits that the pathogenesis of depression lies in overactive liver invading spleen,earth dampness impeding wood′s ascendancy,and disharmony between body and mind,as well as imbalance in storage and discharge functions of liver and kidney,disharmony between Yin and Yang,and disrupted counter-regulation.This dosely aligns with two key pathological methanisms at the micro level:microglial-limbic system homeostatic imbalance and hypothalamic-pituitary-adrenal axis-inflammatory circuit dysregulation.Clinically,the treatment principle for depression adheres to supporting the counter-regulation to restrain excess,with herbal interventions using strategies such as restraining wood to support earth,dredging earth to unblock wood,and harmonizing pivotal functions,as well as nourishing water to nurture wood,warming kidney to tonify liver,and relieving depression to calm the spirit.These approaches aim to regulate the liver,spleen,and kidney,embodying the core therapeutic tenet of"striving for equilibrium,"thereby restoring the body′s self-regulating capability.

Objective:To study the whole-brain distribution of vasoactive intestinal peptide(VIP)-positive neurons in the mouse brain and provide assistance for anatomical and functional studies of VIP neurons.Methods:VIP-Cre::Ai47 mice were used to label VIP neurons in the whole brain.Then,fluorescent imaging of the whole brain slices from VIP-Cre::Ai47 mice was calibrated using the standard brain map.Finally,the distribution density of VIP-positive neurons in different regions of the whole brain was statistically analyzed.Results:The overall distribution densities of VIP neurons in the cortex,olfactory bulb,and hippocampus were all greater than 5 neurons/mm2,and the distribution densities varied greatly in different subregions.The overall distribution density of VIP in the amygdala in the subcortical region was 4 neurons/mm2,and the distribution densities of VIP in the thalamus,hypothalamus,midbrain,and hind-brain regions were less than 2 neurons/mm2 on average,except for that in the supraoptic nucleus of the hypothalamus,where the density of VIP neuron distribution reached 20 neurons/mm2.Conclusion:VIP-positive neurons are mainly distributed in the cortex,hippocampus,and amygdala,which are highly associated with cognition and affection,and are rarely distributed in the thalamus and midbrain.These results suggest that VIP neurons play an essential role in emo-tional and cognitive functions.

Objective:Vascular cell adhesion molecule 1(VCAM1)is involved in a series of physiological and pathological processes,such as immune and inflammatory response,tumor cell metastasis and invasion.But under trau-matic brain injury(TBI),specific types of cells with VC AM1 expression and the related functions are not clear.In order to further explore the specific functions of VCAM1 involved in TBI,this study constructed reporter mice of VCAM1 to explore the response of VCAM1 to TBI in detail.Methods:VCAM1-Cre/ERT2::Ai14 reporter mice were constructed by gene targeting technology and Cre/loxP system,the labeled cell types and labeling efficiency were validated by im-munofluorescence staining and reporter mice hybridization.The stab wound model was used to simulate TBI to induce the changes of VCAM1 expression in cells,and the characteristics of VCAM1 positive astrocytes were detected by immu-nofluorescence staining and fluorescent probe labeling.Results:The labeling efficiency of VCAM1-Cre/ERT2::Ai14 re-porter mice was higher than that of VCAM1 antibody as was seen by labeling of more endothelial cells of blood vessels and unique astrocytes.The distribution of these astrocytes was specific,for example in the nucleus accumbens,amygda-la,hypothalamus,and paraventricular fiber systems.TBI could significantly induce the expression of VCAM1 in astro-cytes(P＜0.0001).These induced astrocytes developed reactive qualities,including somal hypertrophy,GFAP ex-pression and proliferative ability.Conclusion:VCAM1-Cre/ERT2::Ai14 reporter mice could label cells with VCAM1 expression more sensitively,so they were more effective tools for observing expression and function of VCAM1.The up-regulation of VCAM1 expression in astrocytes after TBI surgery suggested that VCAM1 was an inflammatory response molecule in astrocytes,we recommended it as a new molecular indicator of reactive astrocytes.

Objective:To study the whole-brain distribution of vasoactive intestinal peptide(VIP)-positive neurons in the mouse brain and provide assistance for anatomical and functional studies of VIP neurons.Methods:VIP-Cre::Ai47 mice were used to label VIP neurons in the whole brain.Then,fluorescent imaging of the whole brain slices from VIP-Cre::Ai47 mice was calibrated using the standard brain map.Finally,the distribution density of VIP-positive neurons in different regions of the whole brain was statistically analyzed.Results:The overall distribution densities of VIP neurons in the cortex,olfactory bulb,and hippocampus were all greater than 5 neurons/mm2,and the distribution densities varied greatly in different subregions.The overall distribution density of VIP in the amygdala in the subcortical region was 4 neurons/mm2,and the distribution densities of VIP in the thalamus,hypothalamus,midbrain,and hind-brain regions were less than 2 neurons/mm2 on average,except for that in the supraoptic nucleus of the hypothalamus,where the density of VIP neuron distribution reached 20 neurons/mm2.Conclusion:VIP-positive neurons are mainly distributed in the cortex,hippocampus,and amygdala,which are highly associated with cognition and affection,and are rarely distributed in the thalamus and midbrain.These results suggest that VIP neurons play an essential role in emo-tional and cognitive functions.

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.