Role of Bushen Huoxue Formula and transplanted endothelial progenitor cells play in promoting endplate microcirculation and attenuating intervertebral disc degeneration.

Objective: To explore whether Bushen Huoxue Formula (BSHXF) improves the angiogenesis ability of transplanted endothelial progenitor cells (EPCs) in endplate and its potential mechanism in delaying intervertebral disc degeneration (IDD). Methods: BSHXF was analyzed via Ultra-High Performance Liquid Chromatography (UPLC). Rabbit axial compression lumbar IDD models were constructed and the effects of BSHXF, EPCs, and their combination in IDD were determined by MRI, histological evaluation, TUNEL, and immunofluorescence assays. Additionally, CCK-8 assay, flow cytometry, and tube formation assay were used to evaluate EPCs viability, proliferation, cell cycle and the angiogenesis ability of EPCs between groups. Results: BSHXF and transplanted EPCs both attenuate the process of IDD in the rabbit model assessed by MRI, HE staining and Masson staining. TUNEL-positive NP cells were significantly reduced in the BSHXF group, EPCs group, and EPC + BSHXF group compared to the model group (P < 0.05), with the EPC + BSHXF group showing the most significant therapeutic effect. Immunofluorescence detection showed that VEGF, CD34 expression and quantity of microvessels in the endplate significantly increased in the EPC + BSHXF group compared to all the other groups (P < 0.05). Besides, the CCK-8 assay showed an upregulation of EPC viability and the tube formation assay demonstrated a significant increase in tube length and branching in EPCs cultured with BSHXF-containing serum (P < 0.05). Furthermore, BSHXF-containing serum increased VEGF expression in EPCs cultured in vitro (P < 0.05). Conclusions: Both BSHXF and EPCs transplantation play an important role in increasing endplate angiogenesis and attenuating IDD. BSHXF can enhance the viability and tube-forming ability of EPCs and endplate microcirculation.

Network Pharmacology with Metabolomics Study to Reveal the Mechanisms of Bushen Huoxue Formula in Intervertebral Disc Degeneration Treatment.

Background: Intervertebral disc degeneration (IVDD) is a pathophysiological process that leads to severe back pain or neurological deficits. The Bushen Huoxue Formula (BSHXF) is a traditional herbal remedy widely used to treat diseases related to IVDD. However, its pharmacological mechanism needs further exploration. Objective: This study aimed to elucidate the mechanisms through which BSHXF treats IVDD-related diseases by integrating metabolomics with network pharmacology. Methods: Network pharmacology was utilized to identify potential targets of BSHXF against IVDD. Additionally, an animal model of needle puncture-induced disc degeneration was established to assess the effect of BSHXF. Mice were randomly assigned to the sham group, model group, and BSHXF group. Various techniques, including PCR, CCK-8 assay, MRI, histological examinations, and immunohistochemical analyses, were employed to evaluate degenerative and oxidative stress conditions in mouse disc tissue and cultured nucleus pulposus (NP) cells. UHPLC-HRMS/MS was used to differential distinct metabolites in the disc tissue from different groups, and MetaboAnalyst 5.0 was employed to enrich the metabolic pathways. Results: Through network pharmacology, 15 core proteins were identified through protein-protein interaction (PPI) network construction. Functional enrichment analysis highlighted the critical role of BSHXF in addressing IVDD by influencing the response to oxidative stress. Furthermore, experimental evidence demonstrated that BSHXF significantly improved the pathological progression of IVDD and increased oxidative stress markers SOD-1 and GPX1, both in the disc degeneration model and cultured NP cells. Metabolomics identified differential metabolites among the three groups, revealing 15 metabolic pathways between the sham and model groups, and 13 metabolic pathways enriched between the model and BSHXF groups. Conclusion: This study, integrating network pharmacology and metabolomics, suggests that BSHXF can alleviate IVDD progression by modulating oxidative stress. Key metabolic pathways associated with BSHXF-mediated reduction of oxidative stress include the citrate cycle, cysteine and methionine metabolism, alanine, aspartate and glutamate metabolism, glycine, serine and threonine metabolism, D-glutamine and D-glutamate metabolism, glutathione metabolism, and tryptophan metabolism. While this research demonstrates the therapeutic potential of BSHXF in reducing oxidative stress levels in IVDD, further research is needed to thoroughly understand its underlying mechanisms.

A randomized controlled trial: The efficacy and safety of Bushen Huoxue formula in the management of lower back pain from lumbar disc herniation.

BACKGROUND: Lower back pain (LBP) arising from lumbar disc herniation (LDH) poses a challenging health issue, often necessitating therapeutic interventions. Bushen Huoxue formula (BSHXF) has proved as a potential treatment option with great clinical effect. However, comprehensive investigations into its efficacy and safety in conjunction with celecoxib for managing LBP from LDH are lacking. The objective of this article is to investigate the efficacy and safety of BSHXF in the management of patients with LBP from LDH. METHODS: This single center, randomized clinical trial was conducted from March 2023 to September 2023 and all patients suffered from LBP of LDH. Participants were randomly assigned to the BSHXF group (celecoxib and BSHXF) or the control group (celecoxib and placebo). The patients received treatment for 2 weeks. Assessment was conducted before treatment, the last day of the treatment, 4 weeks and 8 weeks after the treatment. Oswestry Disability Index (ODI), Visual Analog Scale (VAS), Roland-Morris Disability Questionnaire (RMDQ), Timed up and go test (TUGT), trunk range of movement (Trunk ROM), Hospital Anxiety and Depression Scale (HADS) were used for the evaluation. RESULTS: A total of 206 subjects completed treatment, among whom 104 participants were randomized to the BSHXF group and 102 participants were randomized to the control group. There were no significant differences between groups in terms of the observed indicators (P > .05). After treatment, patients in BSHXF group obtained significant lower scores at 2-week, 4-week, 8-week of VAS, ODI, RMDQ, TUGT, Trunk ROM and HADS than the baseline data (P < .05). The ODI score was significantly lower than the control group at 2-week, 4-week, 8-week (2w: 11.30 ±â€…5.80 vs 14.23 ±â€…6.33, P < .001; 4w: 10.95 ±â€…4.93 vs 13.54 ±â€…6.35, P < .001; 8w: 10.27 ±â€…5.25 vs 12.84 ±â€…6.57, P = .002). Similarly, the scores of VAS, RMDQ, TUGT, Trunk ROM scores of the BSHXF group markedly decreased at 2, 4, and 8-week when compared to their control group (P < .05). Furthermore, no significant difference showed up in the score of HADS between the between the BSHXF and the control group after treatment (P > .05). CONCLUSION: This randomized clinical trial found that BSXHF can help significantly improve the clinical outcomes of celecoxib including pain intensity reduction and lumbar function improvement in LBP patients.

A Speech Recognition Method Based on Domain-Specific Datasets and Confidence Decision Networks.

This paper proposes a speech recognition method based on a domain-specific language speech network (DSL-Net) and a confidence decision network (CD-Net). The method involves automatically training a domain-specific dataset, using pre-trained model parameters for migration learning, and obtaining a domain-specific speech model. Importance sampling weights were set for the trained domain-specific speech model, which was then integrated with the trained speech model from the benchmark dataset. This integration automatically expands the lexical content of the model to accommodate the input speech based on the lexicon and language model. The adaptation attempts to address the issue of out-of-vocabulary words that are likely to arise in most realistic scenarios and utilizes external knowledge sources to extend the existing language model. By doing so, the approach enhances the adaptability of the language model in new domains or scenarios and improves the prediction accuracy of the model. For domain-specific vocabulary recognition, a deep fully convolutional neural network (DFCNN) and a candidate temporal classification (CTC)-based approach were employed to achieve effective recognition of domain-specific vocabulary. Furthermore, a confidence-based classifier was added to enhance the accuracy and robustness of the overall approach. In the experiments, the method was tested on a proprietary domain audio dataset and compared with an automatic speech recognition (ASR) system trained on a large-scale dataset. Based on experimental verification, the model achieved an accuracy improvement from 82% to 91% in the medical domain. The inclusion of domain-specific datasets resulted in a 5% to 7% enhancement over the baseline, while the introduction of model confidence further improved the baseline by 3% to 5%. These findings demonstrate the significance of incorporating domain-specific datasets and model confidence in advancing speech recognition technology.

A lumbar disc degeneration model established through an external compressive device for the study of microcirculation changes in bony endplates.

Objective: To establish a new animal model of intervertebral disc degeneration (IDD) by axial compression on lumbar spine of rabbits and to investigate the changes of microcirculation in bony endplates during the progress of IDD. Methods: 32 New Zealand white rabbits were equally divided into 4 groups as follows: Control group with no operation and compression, Sham operation group with apparatus placement only, 2-week compression group and 4-week compression group with the devices installed and compressed for their preset duration. All groups of rabbits underwent MRI, histological evaluation, disc height index measurement and Microfil contrast agent perfusions to examine the ratio of endplate microvascular channels. Results: The new animal model of IDD was successfully established after axial compression for 4 weeks. The MRI grades for the 4-week compression group was 4.63 ± 0.52 and significantly different to the sham operation group (P < 0.05). Histologically, decrease of normal NP cells and extracellular matrix and disorganization of the architecture of the annulus fibrosus apparently occurred in 4-week compression group, which was different to the sham operation group (P < 0.05). There was no statistically difference between the 2-week compression and sham operation group no matter in the histology and MRI assessment. The disc height index slowly decreased as the compression duration rose. The ratio of microvascular channel volume within the bony endplate in 2-week and 4-week compression group were both reduced whereas the 4-week compression group obtained significantly less vascularization volume (6.34 ± 1.52 vs. 19.52 ± 4.63, P < 0.05). Conclusion: A new model of lumbar IDD was successfully established by axial compression and the volume of microvascular channels in the bony endplate gradually decreased as the grade of IDD increased. This model provides a new choice for etiological studies on IDD and investigation of nutrient supply disturbance.

CircRNA GRB10 is a Novel Biomarker for the Accurate Diagnosis of Lumbar Degenerative Disc Disease.

Lumbar degenerative disc disease (LDDD) is frequently misdiagnosed as other spine conditions, and the accurate diagnosis is challenging. This study was conducted to explore the role of circRNA GRB10 in the accurate diagnosis of LDDD. This study included 60 cases of LDDD, 60 cases of patients with sacroiliac joint pain (SJP), 60 cases of lumbar disc herniation (LDH), 60 cases of piriformis syndrome (PS), 60 cases of entrapment neuropathy (EN) and 60 cases of healthy controls (HCs). Plasma was obtained from each patient before and after treatment. Expression of GRB10 was studied with RT-qPCR. The role of plasma GRB10 in the accurate diagnosis of LDDD was analyzed by ROC curve analysis. Compared to HCs, decreased accumulation of GRB10 RNA was only observed in LDDD group, but not in SJP, LDH, PS and EN groups. With plasma expression level of GRB10 measured before treatment as a biomarker, LDDD patients were separated from SJP, LDH, PS, EN and HC groups. After treatment, increased expression levels of GRB10 were only observed in LDDD group, but not in other groups. Therefore, GRB10 was downregulated in LDDD and may serve as a biomarker for the accurate diagnosis of LDDD.

Balloon kyphoplasty combined with posterior pedicle screw fixation for the treatment of osteoporotic thoracolumbar burst fractures.

BACKGROUND: Osteoporotic thoracolumbar burst fracture (TLBF) is difficult to treat due to its high rate of postoperative implant failure. This study was designed to evaluate the clinical effect of balloon kyphoplasty with polymethylmethacrylate (PMMA) and posterior pedicle screw fixation for the treatment of osteoporotic TLBF. METHODS: Between February 2012 and May 2016, 24 consecutive patients with osteoporotic TLBFs were included in this study, all of whom suffered incomplete neurologic deficit. They were managed with transpedicular balloon kyphoplasty, PMMA augmentation, and posterior pedicle screw fixation. Scanning with X-ray and computed tomography (CT) were performed to evaluate the kyphotic deformity with the Cobb angle and vertebral body height loss (VBHL) of the injured vertebra. Visual analogue scale (VAS) and the Oswestry Disability Index (ODI) were adopted to assess the pain and dysfunction levels before and after the operation. RESULTS: All participants were followed up for an average of 18 months and those with incomplete neurologic deficit recovered completely. The Cobb angle significantly improved from a preoperative angle of 23.2°±3.6° to 5.3°±2° after operation and to 5.7°±2.2° at the last follow up (P<0.05). The VBHL improved from 56.8%±7.8% before operation to 9.1%±1.6% after operation and to 9.7%±1.9% at the last follow up (P<0.05). The visual analog scale (VAS) score decreased from 8.8±0.9 before operation to 2.5±0.4 after operation and to 1.4±0.4 at the last follow up (P<0.05). The ODI score decreased from 88.6%±3.5% before the operation to 32.3%±3.7% after operation and to 17.5%±1.8% at the last follow up (P<0.05). CONCLUSIONS: Balloon kyphoplasty combined with posterior pedicle screw fixation was an effective treatment for osteoporotic TLBFs. This procedure can reconstruct 3 spinal columns using a single approach with less blood loss, short operation time, and rapid recovery. Ruptures of the posterior vertebral wall should not be a contraindication of this procedure. TRIAL REGISTRATION: Chinese Clinical Trial Registry (NO.: ChiCTR1900026157).

Radiological and clinical outcomes of balloon kyphoplasty for osteoporotic vertebral compression fracture in patients with rheumatoid arthritis.

BACKGROUND: This study was conducted to investigate the outcomes and complications of balloon kyphoplasty (KP) for the treatment of osteoporotic vertebral compression fracture (OVCF) in patients with rheumatoid arthritis (RA) and compare its radiological and clinical effects with OVCF patients without RA. METHODS: Ninety-eight patients in the RA group with 158 fractured vertebrae and 114 patients in the control group with 150 vertebrae were involved in this study. Changes in compression rate, local kyphotic angle, visual analog scale (VAS) and Oswestry disability index (ODI) scores, conditions of bone cement leakage, refracture of the operated vertebrae, and new adjacent vertebral fractures were examined after KP. In addition, patients in the RA group were divided into different groups according to the value of erythrocyte sedimentation rate (ESR), c-reactive protein (CRP), and whether they were glucocorticoid users or not to evaluate their influence on the outcomes of KP. RESULTS: KP procedure significantly improved the compression rate, local kyphotic angle, and VAS and ODI scores in both RA and control groups (p<0.05). Changes in compression rate and local kyphotic angle in the RA group were significantly larger than that in the control group (p<0.05), and patients with RA suffered more new adjacent vertebral fractures after KP. The outcomes and complications of KP from different ESR or CRP groups did not show significant differences. The incidence of cement leakage in RA patients with glucocorticoid use was significantly higher than those who did not take glucocorticoids. In addition, RA patients with glucocorticoid use suffered more intradiscal leakage and new adjacent vertebral fractures. CONCLUSIONS: OVCF patients with RA obtained more improvement in compression rate and local kyphotic angle after KP when compared to those without RA, but they suffered more new adjacent vertebral fractures. Intradiscal leakage and new adjacent vertebral fractures occurred more in RA patients with glucocorticoid use. TRIAL REGISTRATION: Retrospectively registered.

Structural insight into the type-specific epitope of porcine circovirus type 3.

The recently identified pathogenic Porcine circovirus type 3 (PCV3) may threaten to reduce the pig population dramatically worldwide. In our previous study, a PCV3-specific monoclonal antibody (mAb-1H11) was successfully applied in immune-histochemistry staining and ELISA, which specifically recognize PCV3 capsid protein in PCV3-positive pig tissues. In the present study, we expressed and purified the soluble sole capsid protein of PCV3. The purified capsid protein was capable of self-assembly into virus-like-particles (VLPs), which is validated by transmission electron microscopy and dynamic light scattering assays. Moreover, the epitope of mAb-1H11 was identified in the CD-loop region (a.a. 72-79) on the VLP surface, which is confirmed by PCV2-PCV3 epitope swapping assay. For the first time, we determined the cryo-EM structure of PCV3-VLP at 8.5 Å resolution that reveals the detailed structural information of PCV3-VLP. In our cryo-EM structure, PCV3-VLP is composed of 60 capsid protein subunits assembled with T = 1 icosahedral symmetry. Consistent to our bio-dot Western blot assay, the structural comparison between PCV3 and PCV2 revealed significant structural differences in the surface-exposed loops, including the CD-loop (a.a. 72-79) and the EF-loop (a.a. 109-131). Our work provides a structural framework for engineering future PCV3 vaccine and diagnosis kits development.

Loganin alleviates LPS-activated intestinal epithelial inflammation by regulating TLR4/NF-κB and JAK/STAT3 signaling pathways.

Inflammatory bowel disease (IBD) is a chronic, recurrent gastrointestinal inflammation that affects millions of people around the world. Loganin, an iridoid glycoside, has shown the anti-inflammatory effects. However, the effect of loganin on IBD and its underlying molecular mechanism are not clear. The present study aimed to investigate whether loganin could alleviate IBD and its mechanisms. The intestinal epithelial Caco-2 cell line was treated with lipopolysaccharide (LPS) to establish an in vitro IBD model. MTT assay was used to detect cell viability. The expression and release level of inflammatory factors were determined by both real-time-PCR and ELISA. Western blotting was used to assess the NF-κB and JAK/STAT3 pathway-related protein levels. The results showed that loganin repressed the expression and release of IL-6, TNF-α, and IL-1ß, and inhibited TLR4/NF-κB and JAK/STAT3 signaling pathways in a concentration-dependent manner. Overexpression of TLR4 could reverse the effect of loganin, leading to activation of NF-κB signaling and production of inflammatory factors. Meanwhile, IGF-1, a JAK/STAT3 signaling activator, could also reverse the anti-inflammation effect of loganin. In conclusion, loganin inhibited LPS-activated intestinal epithelial inflammation by repressing TLR4/NF-κB and JAK/STAT3 signaling pathway.

The clinical effect of percutaneous kyphoplasty for the treatment of multiple osteoporotic vertebral compression fractures and the prevention of new vertebral fractures.

This study aimed to investigate the clinical effect of percutaneous kyphoplasty and the precautions against adjacent vertebral refractures in the treatment of multiple osteoporotic vertebral compression fractures. 54 cases (128 vertebrae) with multiple osteoporotic vertebral compression fractures from July 2007 to December 2013 treated with percutaneous kyphoplasty were retrospectively reviewed. 36 cases of them suffered from bi-segment vertebral fractures, 16 cases with tri-segment vertebral fractures and 2 cases with quadri-segment vertebral fractures. The operative effect was evaluated by visual analogue scale (VAS) score and oswestry disability index (ODI) score. Then the reasons for adjacent vertebral refractures were analyzed and the precautions were proposed. 54 cases (128 vertebrae) were admitted with percutaneous kyphoplasty successfully. No pulmonary embolism, spinal cord injury and other serious complications were found. The follow-up took 3-33 months with the average of 12 months. There was significant difference of VAS scores and ODI scores between pre-operation and post-operation (P<0.05). Bone cement leakage occurred in 23 vertebrae, and the incidence rate was 18.0%. 8 cases sustained adjacent vertebral refractures including 3 cases in the contiguous vertebral bodies and 5 cases in the interval vertebral bodies, and the incidence rate was 14.8%. 5 cases gained fracture healing after additional percutaneous kyphoplasty procedures while the other 3 cases were healed basically after conservative treatment for three months. In conclusion, percutaneous kyphoplasty is safe and effective to treat multiple osteoporotic vertebral compression fractures. However, the risk of new adjacent vertebral fractures in the multiple osteoporotic vertebral compression fractures is higher than that in the single osteoporotic vertebral compression fracture. Timely and proper treatment can reduce refractures.

The clinical application and efficacy of percutaneous kyphoplasty via unilateral pedicular approach guided by CT image measurement.

This study aimed to investigate the clinical application and efficacy of percutaneous kyphoplasty via unilateral pedicular approach with the reference of preoperative CT image data. 73 cases (a total of 112 vertebrae) with thoracic and lumbar osteoporotic vertebral compression fractures (OVCF) received in our department were collected in this study and underwent percutaneous kyphoplasty via unilateral pedicular approach directed by CT image measurement with the operative time and fluoroscopic times recorded. Visual analogue scale (VAS) and Oswestry disability index (ODI) were used to assess the pain status and functional activity before and after operation and at the last follow-up, while X-ray and CT image were used to measure the height of the injured spinal middle column and kyphotic Cobb angle before and after operation. 73 cases (112 vertebrae) underwent percutaneous kyphoplasty successfully. Cement leakage occurred in 7 cases without obvious neurological symptoms, and they were followed up for 10 to 60 months with the average of 23 months; adjacent vertebral refractures occurred in 3 cases during the follow-up, and their symptoms were relieved after the second surgery. There was significant difference in the height of the injured spinal middle column and kyphotic Cobb angle before and after operation (P<0.05); there was significant difference in preoperative and postoperative VAS score and ODI values (P<0.05). Postoperative CT image data showed that puncture paths of the 110 vertebrae were consistent with preoperative ones sketched using the CT image, and the consistent rate of preoperative and postoperative measurement data was 98%. All patients could ambulate with brace within 2 days after operation without serious complications. In conclusion, percutaneous kyphoplasty via unilateral pedicular approach guided by preoperative CT image data is effective in treatment of osteoporotic vertebral compression fractures, and it is convenient and safe with high puncture accuracy, shorter operative time and less radiation exposure for patients and operators.

[Determination of oleuropein in stem of Syringa oblata from different districts by HPLC].

OBJECTIVE: To develop an HPLC method for the determination of oleuropein in Syringa oblata. METHOD: An Aigilent ZORBAX SB- C18 column (4.6 mm x 250 mm, 5 microm) was used. The mobile phase was acetonitrile-water (21 : 79). The flow rate was 1.0 mL x min(-1). The detection wavelenghth was set at 232 nm and the column temperature was 30 degrees C. RESULT: The linear range of oleuropein were from 0.011 62 g x L(-1) to 1.162 g x L(-1). The average recovery was 98.7% with RSD 2.5% (n=9). CONCLUSION: The method is reliable, accurate and specific. It can be used for quality control of the stem of Syringa oblata.

[Experimental studies on treatment of depression with YJ-XCC1Z3 in mouse models].

OBJECTIVE: To evaluate the pre-clinical effect of YJ-XCC1Z3 on the treatment of depression with the mice mouse. METHOD: YJ-XCC1Z3 was administered at the dose of 405 mg x kg(-1) and 135 mg x kg(-1) to observe the locomotor activity with the mouse locomotor activity recorder apparatus, to observe the effect of YJ-XCC1Z3 on the duration of immohility in the mouse forced swimming test and tail suspension test, to observe the effect of YJ-XCC1Z3 on the body temperature and the metabolism of monoamine neurotransmitters in mouse brain in the mouse model of reserpine induced hypothermia, and to observe the effect of YJ-XCC1 Z3 on the times of 5-HTP induced head-twitches in mice. RESULT: There were no significant changes in the locomotor activity, but a significant reduction in the immobility time was observed in the mice treated with YJ-XCC1Z3 405 mg x kg(-1) and imipramine in the forced swimming test and the tail suspension test. YJ-XCC1Z3 135 mg x kg(-1) and 405 mg x kg(-1) could improve the range of reserpine induced hypothermia in mice, and the latter could also enhance the times of 5-HTP induced head-twitches in mice. YJ-XCC1Z3 405 mg x kg(-1) and 135 mg x kg(-1) could increase the content of 5-HT and NE and decrease the ratio of 5-HIAA/5-HT in mouse brain, but the dose of 405 mg x kg(-1) could decrease the content of DA. The dose of 405 mg x kg(-1) could increase the content of 5-HIAA and had no obvious effect on the content of HVA and DOPAC. CONCLUSION: YJ-XCC1Z3 shows potent antidepressant effect by improving the behaviour of the mouse in depression and not inducing hyperlocomotion in the mice. This effect results in the increase of the content of 5-HT and NE in the mouse brain. YJ-XCC1Z3 can decrease the metabolism of 5-HT to effect the content of 5-HT.