ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism. MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2. ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05). ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.

Objective: To investigate the effects of "liver-smoothing and spirit-regulating" acupuncture on the intestinal flora, lipopolysaccharide (LPS) and the hippocampal toll-like receptor 4 (TLR4)/ transcription factor (NF)-κB signaling pathway in depressive disorder mouse model, and to explore its underlying mechanisms. Methods: Eighteen male SPF-grade C57BL/6J mice were randomly assigned to the control, model, and acupuncture groups using a random number method, with six mice in each group. The depression disorder model was induced in mice from both the acupuncture and model groups using CUMS. The mice in the acupuncture group were treated with acupuncture at the acupoints of "Baihui" (DU20), "Yintang" (DU29), "Hegu" (LI4), and "Taichong" (LR3) on the 15th day of modeling, with a duration of 20 min per session, once per day, for 2 consecutive weeks. Behavioral differences were assessed using the sucrose preference test, open field test, and forced swim test. Hematoxylin-eosin staining was used to observe pathological changes in the hippocampus and colon. The levels of the inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and LPS in the hippocampus and colon were measured using Enzyme-linked Immunosorbent Assay. Western blotting was used to detect the expression of TLR4 and NF-κB protein in the hippocampus. Changes in gut microbiota structure and abundance were analyzed by 16 S rDNA sequencing. Results: Compared to the control group, the model group showed reduced sucrose preference rate, time in the center area, and total distance, with an increase in immobility time (P<0.01). Inflammatory pathological changes were observed in the hippocampal CA1 region and colon. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon increased (P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were increased (P<0.01). The Chao1 index was increased (P<0.01). The relative abundances of Pseudomonadales, Acinetobacter, Moraxellaceae, Solibacillus, Escherichia_shigella, Enterobacteriaceae, Enterobacterales, Dubosiella, and Erysipelottichales were decreased, while the relative abundances of Alloprevotella and gram_negative_bacteriurh_cTPY_13 were increased (P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were upregulated, and the pathway of terpenoid backbone biosynthesis was downregulated (P<0.01). Compared to the model group, the acupuncture group showed increased sucrose preference, time in the center area, and total distance, with a decrease in immobility time (P<0.01). The inflammatory pathological changes in the hippocampal CA1 region and colon were alleviated. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon were reduced(P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were reduced (P<0.01). The Chao1 index was decreased (P<0.05), and the relative abundances of Dubosiella and Erysipelotrichaceae were increased, while the relative abundance of Rikenellaceae, Alloprevotella, and gram_negative_bacteriuch_cTPY_13 were decreased(P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were significantly downregulated, and the pathway of terpenoid backbone biosynthesis was upregulated (P<0.01). Conclusion " Liver-smoothing and spirit-regulating" acupuncture can improve depressive symptoms in depressive disorder mice, potentially through regulating the LPS and TLR4/NF-κB signaling pathway mediated by intestinal flora, reducing the inflammatory response of the hippocampus, and improving the pathological injury of the hippocampus.

BACKGROUND:Stage Ⅱ Kümmell's disease has traditionally been treated with percutaneous kyphoplasty,but this approach is associated with a high incidence of complications such as poor postoperative pain relief,suboptimal cement dispersion,and adjacent vertebral fractures.Studies have shown that cement augmentation of the injured vertebra combined with posterior spinal canal decompression and short-segment fixation has a good effect on the treatment of Kümmell's disease with neurological symptoms.OBJECTIVE:To compare the outcomes of cement-augmented short-segment percutaneous pedicle screw fixation with those of percutaneous kyphoplasty for the treatment of stage Ⅱ Kümmell's disease.METHODS:From January 2020 to January 2023,a total of 49 patients with stage Ⅱ Kümmell's disease from Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine were included in this study,with 15 males and 34 females.According to the treatment method,the patients were divided into the trial group(n=23)and the control group(n=26).The patients in the trial group received cement-augmented short-segment percutaneous pedicle screw fixation,and the patients in the control group received percutaneous kyphoplasty.The postoperative complications were recorded,and the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra were compared between the two groups at 1,6,12 weeks,6,and 12 months after surgery.The Oswestry disability index and lumbar visual analog score were compared at 1 week and 12 months after surgery.RESULTS AND CONCLUSION:(1)All patients in the two groups were followed up for more than 12 months after surgery.Five patients in the control group had adjacent vertebral fractures,three patients had severe kyphosis,and one patient in the trial group had postoperative incision complications.(2)Compared with preoperative data,the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra in both groups were significantly improved after surgery(P＜0.05).The spinal Cobb angle of the trial group was lower than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05),and the ratio of the anterior edge height of the injured vertebra in the trial group was higher than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05).(3)Compared with preoperative data,the Oswestry disability index and lumbar visual analog scale score of the two groups were significantly improved after surgery(P＜0.05).The Oswestry disability index and lumbar visual analog scale score of the trial group were lower than those of the control group at 1 week and 12 months after surgery(P＜0.05).(4)The results show that compared with percutaneous kyphoplasty,cement-augmented short-segment percutaneous pedicle screw fixation for stage Ⅱ Kümmell's disease can better restore the height of the affected vertebra,maintain the shape of the affected vertebra,improve spinal function,and alleviate lumbar pain.

Immunogenic cell death(ICD)is a form of cell death that can activate the immune system,especially in the treatment of cancer.ICD can enhance the recognition of tumors by the immune system and the release of damage associated molecular patterns(DAMPs),to achieve tumor cell death.Oncolytic viruses(OVs)can selectively infect and kill tumor cells without damaging normal cells.OVs are type Ⅱ ICD inducers that induce ICD in tumor cells by targeting the endoplasmic reticulum.Here,we review the characteristics of ICD and the mechanism of ICD induction by OVs.We also review the latest clinical progress involving ICD and discuss future treatment strategies for tumors.

BACKGROUND:Stage Ⅱ Kümmell's disease has traditionally been treated with percutaneous kyphoplasty,but this approach is associated with a high incidence of complications such as poor postoperative pain relief,suboptimal cement dispersion,and adjacent vertebral fractures.Studies have shown that cement augmentation of the injured vertebra combined with posterior spinal canal decompression and short-segment fixation has a good effect on the treatment of Kümmell's disease with neurological symptoms.OBJECTIVE:To compare the outcomes of cement-augmented short-segment percutaneous pedicle screw fixation with those of percutaneous kyphoplasty for the treatment of stage Ⅱ Kümmell's disease.METHODS:From January 2020 to January 2023,a total of 49 patients with stage Ⅱ Kümmell's disease from Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine were included in this study,with 15 males and 34 females.According to the treatment method,the patients were divided into the trial group(n=23)and the control group(n=26).The patients in the trial group received cement-augmented short-segment percutaneous pedicle screw fixation,and the patients in the control group received percutaneous kyphoplasty.The postoperative complications were recorded,and the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra were compared between the two groups at 1,6,12 weeks,6,and 12 months after surgery.The Oswestry disability index and lumbar visual analog score were compared at 1 week and 12 months after surgery.RESULTS AND CONCLUSION:(1)All patients in the two groups were followed up for more than 12 months after surgery.Five patients in the control group had adjacent vertebral fractures,three patients had severe kyphosis,and one patient in the trial group had postoperative incision complications.(2)Compared with preoperative data,the spinal Cobb angle and the ratio of the anterior edge height of the injured vertebra in both groups were significantly improved after surgery(P＜0.05).The spinal Cobb angle of the trial group was lower than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05),and the ratio of the anterior edge height of the injured vertebra in the trial group was higher than that of the control group at 1,6,12 weeks,6,and 12 months after surgery(P＜0.05).(3)Compared with preoperative data,the Oswestry disability index and lumbar visual analog scale score of the two groups were significantly improved after surgery(P＜0.05).The Oswestry disability index and lumbar visual analog scale score of the trial group were lower than those of the control group at 1 week and 12 months after surgery(P＜0.05).(4)The results show that compared with percutaneous kyphoplasty,cement-augmented short-segment percutaneous pedicle screw fixation for stage Ⅱ Kümmell's disease can better restore the height of the affected vertebra,maintain the shape of the affected vertebra,improve spinal function,and alleviate lumbar pain.

Thoracolumbar trauma, including fractures, dislocations and spinal cord injuries, often result from high-energy injuries such as traffic accidents and falls from heights. It not only causes severe pain and restricted movement for patients, but also leads to neurological damage and even permanent disability. Currently, the diagnosis and treatment of thoracolumbar trauma are faced with many problems, such as possible missed diagnosis and misdiagnosis, lack of individualized and standardized treatment plans, and lack of objective and quantitative metrics for postoperative assessment. Artificial intelligence (AI) technology offers innovative ideas to these problems. Among them, the core AI technology such as machine learning (ML), deep learning (DL), computer vision, and robotics has demonstrated outstanding capabilities in medical image analysis, clinical decision support, etc., which can significantly improve the diagnostic precision, surgical planning efficiency, and postoperative management level of thoracolumbar trauma. At present, application of AI technology in cross-modal data integration, clinical decision support, and long-term efficacy prediction in the field of thoracolumbar trauma remains to be systematically sorted out. To this end, the authors reviewed the research progress of AI technology in the diagnosis, treatment, and postoperative management of thoracolumbar trauma, providing a reference for a wide application of AI technology in the management of thoracolumbar trauma.

Immunogenic cell death(ICD)is a form of cell death that can activate the immune system,especially in the treatment of cancer.ICD can enhance the recognition of tumors by the immune system and the release of damage associated molecular patterns(DAMPs),to achieve tumor cell death.Oncolytic viruses(OVs)can selectively infect and kill tumor cells without damaging normal cells.OVs are type Ⅱ ICD inducers that induce ICD in tumor cells by targeting the endoplasmic reticulum.Here,we review the characteristics of ICD and the mechanism of ICD induction by OVs.We also review the latest clinical progress involving ICD and discuss future treatment strategies for tumors.

Thoracolumbar trauma, including fractures, dislocations and spinal cord injuries, often result from high-energy injuries such as traffic accidents and falls from heights. It not only causes severe pain and restricted movement for patients, but also leads to neurological damage and even permanent disability. Currently, the diagnosis and treatment of thoracolumbar trauma are faced with many problems, such as possible missed diagnosis and misdiagnosis, lack of individualized and standardized treatment plans, and lack of objective and quantitative metrics for postoperative assessment. Artificial intelligence (AI) technology offers innovative ideas to these problems. Among them, the core AI technology such as machine learning (ML), deep learning (DL), computer vision, and robotics has demonstrated outstanding capabilities in medical image analysis, clinical decision support, etc., which can significantly improve the diagnostic precision, surgical planning efficiency, and postoperative management level of thoracolumbar trauma. At present, application of AI technology in cross-modal data integration, clinical decision support, and long-term efficacy prediction in the field of thoracolumbar trauma remains to be systematically sorted out. To this end, the authors reviewed the research progress of AI technology in the diagnosis, treatment, and postoperative management of thoracolumbar trauma, providing a reference for a wide application of AI technology in the management of thoracolumbar trauma.

Objective To investigate the effects of microRNA(miRNA,miR)-887-3p on the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells and its underlying molecular mechanism.Methods Annulus fibrosus tissues were obtained from 8-week-old SPF-grade SD male rats,centrifuged to prepare and identify annulus fibrosus cells.Rats in the experiment were randomly divided into four groups:a Normal group consisting of primary annulus fibrosus cells without any treatment;a Control group treated with 10 ng/mL interleukin-1β(IL-1β)for 24 hours to establish a degenerative cell model;an interference group(miR-887-3p inhibitor)transfected with miR-887-3p inhibitor using Lipo3000 based on the Control group;and an overexpression group(miR-887-3p mimics)transfected with miR-887-3p mimics using Lipo3000 based on the Control group.CCK-8 assay was used to assess cell viability;flow cytometry was used to measure cell apoptosis rates;real-time fluorescence quantitative PCR(qPCR)was used to detect the expression levels of miR-887-3p and murine double minute 4(MDM4)mRNA;Western blotting was used to measure the protein expression levels of MDM4,Bcl-2,and Caspase-3.Results Immunofluorescence staining of isolated and cultured cells revealed a Collagen I positive rate of over 90％in rat intervertebral disc annulus fibrosus cells,indicating a cell purity level greater than 90％.Real-time fluorescence qPCR results showed that after establishing an annulus fibrosus degenerative cell model using IL-1β,the expression level of miR-887-3p significantly increased compared to the Normal group(P＜0.001).Compared to the Control group,transfection with miR-887-3p inhibitor resulted in a significant decrease in its expression level(P＜0.001).The CCK-8 assay showed that compared to the Normal group,cell viability significantly decreased in the Control group(P＜0.001).Compared to the Control group,cell proliferation ability significantly increased after miR-887-3p inhibition,and significantly decreased after overexpression of miR-887-3p.Flow cytometry results revealed that compared to the Normal group,the apoptosis rate in the Control group significantly increased(P＜0.001).Compared to the Control group,the cell apoptosis rate significantly decreased in the miR-887-3p interference group(P＜0.001)and increased in the overexpression group(P＜0.001).Western blotting analysis showed that compared to the Normal group,Bcl-2 expression level significantly decreased(P＜0.001)and Caspase-3 expression level significantly increased(P＜0.001)in the Control group.Compared to the Control group,Bcl-2 and MDM4 expression levels significantly increased(P＜0.01),and Caspase-3 expression level significantly decreased(P＜0.01)in the miR-887-3p interference group;whereas in the overexpression group,Bcl-2 and MDM4 expression levels significantly decreased(P＜0.05),and Caspase-3 levels significantly increased(P＜0.05).Real-time fluorescence qPCR and protein immunoblotting results showed that after interfering with miR-887-3p,the expression of MDM4 protein and mRNA increased(P＜0.001);after overexpressing miR-887-3p,their expression decreased(protein,P＜0.01;mRNA,P＜0.001).Conclusion MiR-887-3p may modulate the cell proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells by regulating MDM4 expression,thereby influencing the development and progression of disc degeneration.

Objective:Biological markers of the ventrolateral orbitofrontal cortex(vlOFC)involved in pain regula-tion were screened.Methods:Chronic inflammatory pain was induced in male C57BL/6J mice by injection of complete Freund's adjuvant(CFA)into the left posterior plantar.Paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were detected to evalue hyperalgesia.Transcriptome sequencing was performed on fresh tissue from vlOFC of mice after behavioral tests.The differentially expressed genes(DEGs)were screened by bioinformatics method,and their biological functions and pathways were enriched.Results:Compared with the PBS group,the left hindpaw me-chanical pain threshold and the paw withdrawal latency caused by heat pain were significantly reduced in the CFA group(P＜0.001).The DEGs of vlOFC in the two groups were 497,of which 143 were up-regulated and 354 were down-reg-ulated.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGGs)analysis showed that:In chro-nic inflammatory pain model mice,DEGs of vlOFC were mainly manifested in biological processes such as organic cation transport,neurotransmitter transport,and regulation of cytoplasmic calcium ion concentration.It is related to G protein-coupled receptors(GPCRs),neuropeptides and ammonium transport.DEGs mainly focuses on neuroactive ligand-receptor interactions,cytokine-cytokine receptor interactions,and cAMP signaling pathways.Reactome functional en-richment analysis showed that the pathway with the highest number of DEGs enriched and the lowest P value-adjusted was GPCRs ligand binding.Conclusion:Ion transport,neurotransmitter transport and binding,and GPCRs-related ac-tivities in vlOFC are involved in the regulation of chronic inflammatory pain.