Calycosin promotes axon growth by inhibiting PTPRS and alleviates spinal cord injury.

Our former studies have identified the alleviating effect of Calycosin (CA) on spinal cord injury (SCI). In this study, our purpose is to explore the influence of CA on SCI from the perspective of promoting axon growth. The SCI animal model was constructed by spinal cord compression, wherein rat primary cortex neuronal isolation was performed, and the axonal growth restriction cell model was established via chondroitin sulfate proteoglycan (CSPG) treatment. The expressions of axon regeneration markers were measured via immunofluorescent staining and western blot, and the direct target of CA was examined using silver staining. Finally, the expression of the protein tyrosine phosphatase receptor type S (PTPRS) was assessed using western blot. CA treatment increased neuronal process outgrowth and the expressions of axon regeneration markers, such as neurofilament H (NF-H), vesicular glutamate transporter 1 (vGlut1), and synaptophysin (Syn) in both SCI model rats and CSPG-treated primary cortical neurons, and PTPRS levels were elevated after SCI induction. In addition, PTPRS was the direct target of CA, and according to in vivo findings, exposure to CA reduced the PTPRS content. Furthermore, PTPRS overexpression inhibited CA's enhancement of axon regeneration marker content and neuronal axon lengths. CA improves SCI by increasing axon development through regulating PTPRS expression.

A Retrospective Comparison of Low-Temperature Plasma Ablation and Radiofrequency Thermocoagulation of the Thoracic Nerve Root for Refractory Postherpetic Neuralgia.

BACKGROUND: Radiofrequency thermocoagulation (RFT) of the thoracic nerve root is commonly employed in treating medication-refractory thoracic post-herpetic neuralgia (PHN). However, RFT procedures' suboptimal pain relief and high occurrence of postoperative skin numbness present persistent challenges. Previous single-cohort research indicated that the low-temperature plasma coblation technique may potentially improve pain relief and reduce the incidence of skin numbness. Nevertheless, conclusive evidence favoring coblation over RFT is lacking. OBJECTIVES: To compare the clinical outcomes associated with coblation to those associated with RFT in the treatment of refractory PHN. STUDY DESIGN: Retrospective matched-cohort study. SETTING: Affiliated Hospital of Capital Medical University. METHODS: Sixty-eight PHN patients underwent coblation procedures between 2019 and 2020, and 312 patients underwent RFT between 2015 and 2020 in our department. A matched-cohort analysis was conducted based on the criteria of age, gender, weight, pain intensity, pain duration, side of pain, and affected thoracic dermatome. Pain relief was assessed using the numeric rating scale (NRS), the Medication Quantification Scale (MQS) Version III and the Neuropathic Pain Symptom Inventory (NPSI), which were employed to indicate pain intensity, medication burden, and comprehensive pain remission at 6, 12, and 24 months. Numbness degree scale scores and complications were recorded to assess safety. RESULTS: We successfully matched a cohort of 59 patients who underwent coblation and an equivalent number of patients who underwent RFT as a PHN treatment. At the follow-up time points, both groups' NRS, MQS, and NPSI scores exhibited significant decreases from the pre-operation scores (P < 0.05). The coblation group's NRS scores were significantly lower than the RFT group's at the sixth and the twenty-fourth months (P < 0.05). At 24 months, the MQS values in the coblation group were significantly lower than those in the RFT group (P < 0.05). Furthermore, the coblation group's total intensity scores on the NPSI were significantly lower than the RFT group's at the 12- and 24-month follow-ups (P < 0.05). At 6 months, the coblation group's temporary intensity scores on the NPSI were significantly lower than the RFT group's (P < 0.05). Notably, the occurrence of moderate or severe numbness in the coblation group was significantly lower than in the RFT group at 6 and 12 months (P < 0.05). No serious adverse effects were reported during the follow-up. LIMITATIONS: This analysis was a single-center retrospective study with a small sample size. CONCLUSION: In this matched cohort analysis, coblation achieved longer-term pain relief with a more minimal incidence rate of skin numbness than did RFT. Further randomized controlled trials should be conducted to solidify coblation's clinical superiority to RFT as a PHN treatment.

Analyzing the Effect of Intraoperative Stimulation Voltage on Facial Numbness Following Radiofrequency Thermocoagulation in the Treatment of Idiopathic Trigeminal Neuralgia.

INTRODUCTION: Radiofrequency thermocoagulation (RFT) effectively alleviates idiopathic trigeminal neuralgia (ITN); however, postoperative facial numbness poses a significant challenge. This issue arises due to the close proximity of high-temperature thermocoagulation, which not only ablates pain-related nociceptive fibers but also affects tactile fibers. Intraoperative sensory stimulation voltage (SV), which reflects the distance between the RFT cannula and the target nerve, potentially possesses the ability to prevent tactile fiber injury. This study aimed to investigate the influence of SV on postoperative facial numbness and provide valuable insights to mitigate its occurrence. METHODS: A retrospective analysis was performed on 72 ITN patients with maxillary division (V2) pain who underwent RFT between 2020 and 2022. Among them, 13 patients with SV ≤ 0.2 V constituted the low SV group. Subsequently, a matched-cohort analysis was conducted on the remaining 59 patients. The patients paired with the low SV patients were subsequently enrolled in the high SV group, adhering to a 1:1 match ratio. The primary outcome was the facial numbness scale assessment at 3 days, 3 months and 6 months post-surgery. The pain intensity and medication burden served as the secondary outcomes. RESULTS: We successfully matched a cohort consisting of 12 patients in the low SV group and 12 patients in the high SV group. Each patient experienced various degrees of facial numbness at 3 days post-RFT. Notably, the low SV group exhibited a higher incidence of moderate numbness (66.7% vs. 16.67%, P = 0.036), whereas the high SV group had more cases of mild numbness at the 6-month follow-up (25% vs. 83.3%, P = 0.012). Both groups demonstrated significant decreases in pain intensity and medication burden compared to before the operation. CONCLUSIONS: SV proved to be a reliable parameter for mitigating the degree of postoperative facial numbness in RFT treatment for ITN. A relatively high sensory SV ranging from 0.3 to 0.6 V during the RFT procedure results in less facial numbness in the treatment of ITN.

Calycosin ameliorates spinal cord injury by targeting Hsp90 to inhibit oxidative stress and apoptosis of nerve cells.

BACKGROUND: Zhenbao pill is effective in protecting against spinal cord injury (SCI). We attempt to explore the characteristics of calycosin (a main monomer of Zhenbao pill) in SCI and its relative mechanism. METHODS: The target of calycosin was screened using pharmacological network analysis. The SCI cell model was constructed using hydrogen peroxide (H2O2), and the animal model was developed by compressing spinal cord with a vascular clamp. Flow cytometry was conducted to test reactive oxygen species (ROS) levels and cell apoptosis. Detection of malondialdehyde (MDA) activity and Superoxide dismutase (SOD) activity were performed using relative kits. Heat shock protein 90 (HSP90) was examined using western blot and quantitative real-time PCR. Motor function tests were carried out. The hematoxylin-eosin and Nissl staining were conducted. RESULTS: In SCI models, ROS, MDA, and cell apoptosis were elevated, SOD and HSP90 levels were restrained, while calycosin addition reversed the above results. Besides, calycosin application or HSP90 overexpression enhanced phosphorylation of protein kinase B (Akt) but weakened that of apoptosis signal-regulating kinase 1 (ASK1) and p38, while HSP90 inhibitor 17-AAG treatment restrained the above results. Meanwhile, the injection of calycosin improved the motor function in SCI model rats. Furthermore, the pathologic results also clarified the positive effect of calycosin on SCI. CONCLUSION: HSP90 was lowly expressed in SCI models. Calycosin alleviated SCI by promoting HSP90 up-regulation and inhibiting oxidative stress and apoptosis of nerve cells.

Regulatory mechanism of Scutellaria baicalensis Georgi on bone cancer pain based on network pharmacology and experimental verification.

Context: Scutellaria baicalensis Georgi (SBG) may relieve bone cancer pain (BCP) by regulating cell proliferation, angiogenesis, and apoptosis. Objective: The mechanism of SBG in the treatment of BCP remains to be further explored. Methods: The active compounds and targets of SBG were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction databases. BCP-related targets were screened from NCBI and GeneCards databases. Additionally, Cytoscape software was applied to construct network diagrams, and OmicShare platform was used to enrich Gene Ontology (GO) and pathways. Finally, the verification of active compounds and core targets was performed based on quantitative real-time PCR (qRT-PCR). Results: Interestingly, we identified baicalein and wogonin as the main active components of SBG. A total of 41 SBG targets, including VEGFA, IL6, MAPK3, JUN and TNF, were obtained in the treatment of BCP. In addition, pathways in cancer may be an essential way of SBG in the treatment of BCP. Experimental verification had shown that baicalein and wogonin were significantly related to BCP core targets. Conclusions: The active components of SBG have been clarified, and the mechanism of the active components in treating BCP has been predicted and verified, which provides an experimental and theoretical basis for the in-depth elucidation of the pharmacodynamics material basis and mechanism of SBG.

Transcriptome Sequencing Explores the Mechanism of Baicalin on Bone Cancer Pain.

INTRODUCTION: Bone cancer pain is characterized by persistent pain, usually requiring drugs to relieve pain. Baicalin, a flavonoid compound extracted from Scutellaria baicalensis, which has antioxidant and analgesic effects. But, the effect of baicalin on bone cancer pain is unclear. Thus, this study aimed to explore the mechanism of baicalin on SD rats with bone cancer pain. MATERIALS AND METHODS: The MADB-106 breast cancer cells-induced bone pain model was constructed and carried out baicalin treatment. The therapeutic effect of baicalin on bone cancer pain model was observed by hematoxylin-eosin staining and immunofluorescence staining. We also performed transcriptome sequencing analysis of baicalin in the treatment of bone metastases. Also, RT-qPCR and ELISA were used to detect the expression levels of inflammation factors. RESULTS: After baicalin treatment, osteoclast activation was inhibited and the number of bone trabeculae was increased. Baicalin inhibited the protein expression level of inflammatory factors (IL-1ß, IL-6, TNF-α and PGE2) in the bone metastases group. Based on the transcriptome sequencing of the bone metastases group and the baicalin treatment group, baicalin inhibited the expression of ALPP, DUSP1, CYR61, ALPPL2, SPP1 and TLR4. RT-qPCR was also used to validate the expression levels of these cytokine genes. CONCLUSION: Baicalin had a certain inhibitory effect on the SD rat model of bone metastasis cancer. These insights can guide future research on the molecular mechanism of bone cancer pain and provide a theoretical basis for baicalin in the treatment of bone pain caused by breast cancer in the future.

A Novel Mechanism of BAM8-22 Inhibiting Microglia Activation: Represses CX3CR1 Expression via Upregulating miR-184.

Bone cancer pain (BCP) is the most common type of pain in cancer patients, during which microglia cells were activated. A previous study showed BAM8-22 had the ability to alleviate BCP via inhibiting microglia activation while the mechanism was not clear. This study aims to investigate the specific mechanism of BAM8-22 inhibiting microglia activation. This study was mainly investigated in BCP mice or LPS-treated microglia BV-2 cells. The behavior tests of mice were performed at 0, 1, 2, 12, and 24 h after BAM8-22 treatment. The expression of miR-184 and CX3CR1 mRNAs was detected by quantitative RT-PCR. The expression of CX3CR1 protein and microglia activation marker, Iba-1, was measured by western blot analysis. The levels of TNF-α and IL-1ß were detected by ELISA. Dual-luciferase assay was performed to verify the combination between miR-184 and CX3CR1. After BAM8-22 treatment, increased miR-184 level was observed in both BCP mice and LPS-treated BV-2 cells, with the downregulated expression of Iba-1 and inflammatory cytokines, namely the inhibition of microglia activation. The inhibition of miR-184 reversed the inhibitory effect of BAM8-22 on microglia activation. Further, in vitro studies showed that miR-184 bound to the 3'UTR of CX3CR1 and inhibited microglia activation via repressing CX3CR1 expression. What's more, the suppression of CX3CR1 expression eliminated the reversal effect of the miR-184 inhibitor on BAM8-22-induced microglia activation and decreased Iba-1 expression and pro-inflammatory cytokine secretion. In BCP models, miR-184 was upregulated by BAM8-22 and the elevated level of miR-184 bound to the 3'UTR region of CX3CR1 and repressed CX3CR1 expression, thus inhibiting the microglia activation, suggesting the potential application of miR-184/CX3CR1 for BCP treatment.

POM121 is identified as a novel prognostic marker of oral squamous cell carcinoma.

Background: The aim of this study was to confirm the role of nuclear pore membrane protein 121(POM121) in oral squamous cell carcinoma and to explore the underlying mechanism. Methods: POM121mRNA and protein expressions were evaluated in OSCC tissues and normal oral tissues by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry. The relationship between POM121 expression and clinical characteristics was analyzed. Bioinformatics analysis was performed to explore the possible mechanisms how POM121 affected OSCC. Results: We confirmed that POM121 mRNA expression in OSCC tissues was significantly higher than that in non-tumorous tissues, as was POM121 protein expression. POM121 expression was associated with distant metastasis and TNM stage. Multivariate analysis confirmed POM121 expression as an independent prognostic factor for OSCC patients. OSCC patients with high POM121 expression had a worse overall survival (OS) compared with patients with low POM121 expression. Bioinformatics analysis indicated POM121 may regulate OSCC through hedgehog and /or p53 signaling pathway. Conclusion: Targeting of POM121 expression levels could provide new diagnostic and therapeutic strategies for OSCC patients.

Dexmedetomidine alleviated lipopolysaccharide/D-galactosamine-induced acute liver injury in mice.

Dexmedetomidine (DEX), a highly selective α2-adrenergic receptor (α2-AR) agonist, is widely used as sedative in clinical. Its potential anti-inflammatory properties have been found in recent studies. The current study has investigated the profound effects of DEX on acute liver injury in mice. The mice were intraperitoneally injected lipopolysaccharide (LPS) and D-galactosamine (D-Gal) to induce acute liver injury, and vehicle or DEX were treated 30 min before or 2 h after LPS/D-Gal exposure. The results showed that pre-treatment with DEX inhibited the raising of plasma aminotransferases, reduced the damage of liver tissue, and improved the survival rate in mice exposed to LPS/D-Gal. Pre-treatment with DEX also inhibited the release of TNF-α and suppressed the phosphorylation of c-jun-N-terminal kinase (JNK) in mice exposed to LPS/D-Gal. In addition, pre-treatment with DEX down-regulated the expression of cleavage of caspase-3, decreased the activities of caspase-3, caspase-8, caspase-9, and consequently, reduced hepatocyte apoptosis. Interestingly, post-treatment with DEX also resulted in beneficial outcomes. The current study indicates that administration of DEX might provide protective benefits in inflammatory liver disease.

IP3-Mediated Calcium Signaling Is Involved in the Mechanism of Fractalkine-Induced Hyperalgesia Response.

BACKGROUND Fractalkine is widely expressed throughout the brain and spinal cord, where it can exert effects on pain enhancement and hyperalgesia by activating microglia through CX3C chemokine receptor 1 (CX3CR1), which triggers the release of several pro-inflammatory cytokines in the spinal cord. Fractalkine has also been shown to increase cytosolic calcium ([Ca2+]i) in microglia. MATERIAL AND METHODS Based on the characteristics of CX3CR1, a G protein-coupled receptor, we explored the role of inositol 1,4,5-trisphosphate (IP3) signaling in fractalkine-induced inflammatory response in BV-2 cells in vitro. The effect and the underlying mechanism induced by fractalkine in the brain were observed using a mouse model with intracerebroventricular (i.c.v.) injection of exogenous fractalkine. RESULTS [Ca2+]i was significantly increased and IL-1ß and TNF-α levels were higher in the fractalkine-treated cell groups than in the farctalkine+ 2-APB groups. We found that i.c.v. injection of fractalkine significantly increased p-p38MAPK, IL-1ß, and TNF-α expression in the brain, while i.c.v. injection of a fractalkine-neutralizing antibody (anti-CX3CR1), trisphosphate receptor (IP3R) antagonist (2-APB), or p38MAPK inhibitor (SB203580) prior to fractalkine addition yielded an effective and reliable anti-allodynia effect, following the reduction of p-p38MAPK, IL-1ß, and TNF-α expression. CONCLUSIONS Our results suggest that fractalkine leads to hyperalgesia, and the underlying mechanism may be associated with IP3/p38MAPK-mediated calcium signaling and its phlogogenic properties.

Lipoxin A4 inhibits the production of proinflammatory cytokines induced by ß-amyloid in vitro and in vivo.

Studies increasingly indicate that inflammation induced by ß-amyloid (Aß) contributes to the progression of Alzheimer's disease (AD). How to inhibit the enhanced production of proinflammatory cytokines stimulated by Aß is an important research subject for the treatment of AD. In this study, we investigated the inhibitory effect and the molecular mechanism of the lipoxin A(4) (LXA(4)) on the production of interleukin-1ß (IL-1ß) and tumor necrosis factorα (TNFα) induced by ß-amyloid in the cortex and hippocampus of mice, and in Aß-stimulated BV2 cells, a mouse microglial cell line. LXA(4) down-regulated the protein expression of IL-1ß and TNFα, attenuated the gene expressions of IL-1ß and TNFα, inhibited the degradation of IκBα, inhibited translocation of NF-κB p65 subunit into the nucleus induced by ß-amyloid in the cortex and hippocampus of mice, and in Aß-stimulated BV2 cells, and the inhibitory effects were dose dependently elevated. Our findings suggest that LXA(4) inhibits the production of IL-1ß and TNFα induced by ß-amyloid in the cortex and hippocampus of mice, and in BV2 microglial cells via the NF-κB signal pathway.