Background: Breast cancer stem-like cells (BCSCs) are considered a source of tumor origins, metastasis and drug resistance, thereby limiting current treatment regimens. Stigmasterol has been reported to inhibit various cancer processes, but its effects and mechanisms in BCSCs have not been investigated. Methods: To generate spheroids, we enriched parental and SUM159 cells with BCSCs in a serum-free medium. The effects on the stemness, metastasis and drug resistance of CSC-enriched SUM159 cells were detected for the first time by in vivo and in vitro experiments. Results: CSC-enriched SUM159 and 4T1 cells demonstrated higher potential for tumorigenesis and metastasis. Stigmasterol suppresses BCSCs' spheroid formation, cell viability, and migration ability and promotes cell apoptosis. Stigmasterol also inhibited BCSCs-originated cancer formation in rat models. Stigmasterol also attenuated the growth of TNBC organoids from human breast cancer tissues. These data revealed the inhibitory effects of stigmasterol on BCSC traits. In the meantime, we found that JAK3 was upregulated in BCSCs, and Stigmasterol could effectively inhibit its expression. In addition, JAK3 was evidenced to negatively regulate BCSC activity and stemness both in vitro and in vivo. More importantly, the results indicated that Stigmasterol suppresses BCSC activity by inhibiting JAK3 expression. Conclusion: This study is the first to demonstrate that Stigmasterol inhibited metastasis and stemness of BCSCs by downregulating JAK3, which might provide a new method for the clinical application of Stigmasterol in breast cancer.

Background :The VBQ Score, a formula based on magnetic resonance imaging (MRI) parameters, facilitates the assessment of bone quality in patients with osteoporotic vertebral fractures, offering substantial clinical convenience. However, the formula necessitates lumbar MRI imaging for data collection and evaluation, leading to practical challenges in clinical scenarios. Specifically, in the case of a thoracic vertebral fracture, additional lumbar spine MRI is necessary to obtain pertinent data, or when incomplete lumbar spine MRI images exist, the measurement of relevant data becomes challenging. Purpose: This study introduces a m-VBQ score and compares it with the standard VBQ score, investigating the feasibility and accuracy of the improved VBQ score for evaluating bone quality. Methods: A retrospective study was conducted, involving adult patients from January 2017 to December 2022. The study included a patient sample comprising 103 individuals with osteoporotic vertebral fractures (OVF) and 103 without OVF. METHODS: The collected data encompassed non-contrast T1-weighted MRI, DEXA, and demographic information from both the fracture group and the non-fracture group patients. The m-VBQ score was established by selecting the vertebral scoring site, which could be either the vertebral body of a single-segment fractured vertebra or the vertebral body of a single-segment non-fractured vertebra. This involved calculating the median signal intensity (MSI) of the two vertebrae above and the two vertebrae below the vertebral scoring site, followed by dividing this value by the signal intensity of the cerebrospinal fluid (CSF) in the vertebra immediately below the vertebral scoring site. The differences between the two m-VBQ score groups were evaluated using the Student's t-test. Additionally, the relationship between the m-VBQ score and VBQ score was examined through Pearson correlation analysis, univariate linear regression, and intra-group correlation analysis. Binary logistic regression was utilized to investigate the association of the VBQ score, m-VBQ score, and occurrence of OVF. Receiver Operating Characteristic (ROC) analysis was performed to determine the Area Under the Curve (AUC) for both scores as predictors of OVF and osteoporosis (OP). Furthermore, subgroup analyses were conducted based on the vertebral score location, with statistical significance determined at P<0.05. Results:There was a significant difference in m-VBQ scores between the fracture and non-fracture groups (3.88±0.86 vs. 3.07±0.59, P < 0.001). Pearson correlation analysis revealed a strong positive association between m-VBQ scores and VBQ scores (r=0.842, P<0.001). As the scoring vertebral site approached L2 from above, the Pearson correlation coefficient increased, reaching its peak at L2 (r=0.951, P<0.001) and subsequently decreasing for sites below. Linear regression analysis between m-VBQ and VBQ scores indicated that as the scoring vertebral site approaches L2 from above, the slope of the equation might increase, with the maximum slope observed at L2 (β1=0.9757), and then potentially decrease for lower sites. The consistency between VBQ scores and m-VBQ scores is excellent (ICC = 0.841). As the vertebral scoring site of the m-VBQ score progresses downward, nearing L2, the ICC coefficient potentially increases, reaching its peak at L2 (ICC = 0.950). Beyond this point, the ICC coefficient might decrease.Binary logistic regression highlighted that m-VBQ scores (OR=1.253-5.282, 95%CI:1.210-5.496, P=0.010) had a stronger significance in predicting OVF compared to VBQ scores (OR=2.579, 95%CI:1.210-5.496, P=0.014). Regarding OVF prediction, the AUC for m-VBQ was 0.776 (95%CI:0.713-0.838, P<0.001), indicating good predictive capability, similar to VBQ. For specific vertebral sites, m-VBQ's predictive ability was superior to VBQ for the T3-T11 subgroup, equivalent for the L3-L5 subgroup, and inferior for the T12, L1, and L2 subgroups. In predicting osteoporosis (OP), the AUC for m-VBQ was 0.737 (95%CI:0.669-0.805, P<0.001), suggesting a stronger predictive capacity compared to VBQ. Inter-rater reliability for m-VBQ scores among two reviewers yielded an ICC of 0.896 (95%CI 0.861-0.963, P<0.001), and intra-rater reliability resulted in an ICC of 0.81 (95% CI 0.759-0.853, P<0.001). Conclusion:The VBQ score and the m-VBQ score, derived from the adjacent vertebrae of OVF patients, are both effective indices for assessing bone quality in OVF patients. The m-VBQ score exhibits good predictive capability for OVF and OP, closely mirroring the VBQ score's predictive power. Compared to the conventional VBQ score, we believe the m-VBQ score offers more flexibility and clinical operability while maintaining similar reliability for evaluating OP and OVF. Furthermore, it enables the assessment of bone quality in vertebrae adjacent to fractures, potentially predicting subsequent fractures in neighboring vertebrae.

The sequences of primers used in this study

Ethnopharmacological relevance: Two types of traditional Chinese formulas of botanical drugs are prescribed for treating perimenopausal syndrome (PMS), a disorder in middle-aged women during their transition to menopause. One is for treating PMS as kidney deficiency (KD) due to senescence and declining reproductive functions, and the other is for treating it as liver qi stagnation (LQS) in association with stress and anxiety. Despite the time-tested prescriptions, an objective attestation to the effectiveness of the traditional Chinese treatment of PMS is still to be established and the associated molecular mechanism is still to be investigated. Materials and methods: A model for PMS was generated from perimenopausal rats with chronic restraint stress (CRS). The effectiveness of traditional Chinese formulas of botanical drugs and a combination of two of the formulas was evaluated based on 1H NMR plasma metabolomic, as well as behavioral and physiological, indicators. To investigate whether the formulas contained ligands that could compensate for the declining level of estrogen, the primary cause of PMS, the ligand-based NMR technique of saturation transfer difference (STD) was employed to detect possible interacting molecules to estrogen receptors in the decoction. Results: Each prescription of the classical Chinese formula moderately attenuated the metabolomic state of the disease model. The best treatment strategy however was to combine two traditional Chinese formulas, each for a different etiology, to adjust the metabolomic state of the disease model to that of rats at a much younger age. In addition, this attenuation of the metabolomics of the disease model was by neither upregulating the estrogen level nor supplementing an estrogenic compound. Conclusion: Treatment of PMS with a traditional Chinese formula of botanical drugs targeting one of the two causes separately could ameliorate the disorder moderately. However, the best outcome was to treat the two causes simultaneously with a decoction that combined ingredients from two traditional prescriptions. The data also implicated a new paradigm for phytotherapy of PMS as the prescribed decoctions contained no interacting compound to modulate the activity of estrogen receptors, in contrast to the treatment strategy of hormone replacement therapy.

目的:探讨扶正固本法对乳腺癌骨转移后骨破坏的抑制作用及可能的分子机制.方法:通过胫骨内注射乳腺癌细胞建立乳腺癌骨转移动物模型,给予扶正固本方药干预,给药后采用IVIS检测乳腺癌在骨环境中的生长情况,X-ray和TRAP染色分析骨破坏及破骨细胞活性情况,并采用ELISA检测小鼠血清中的TNF-α、IL-1β、IL-6、IL-17、PTHrP、RANKL、OPG、TGF-β、IGF、PDGF细胞因子的表达改变.结果:IVIS结果显示扶正固本方药可减缓乳腺癌在骨环境中的生长,X-ray和TRAP染色表明扶正固本方药可以减少乳腺癌骨转移引起的骨破坏及破骨细胞活性,ELISA结果显示扶正固本方药可以降低血清中TNF-α、IL-1β、IL-6、IL-17、PTHrP、RANKL、TGF-β、IGF、PDGF的表达水平,提高血清中OPG的表达水平.结论:扶正固本方药可通过调节多种细胞因子的表达,从而减缓乳腺癌细胞在骨环境中的生长,以及减轻乳腺癌细胞所引起的骨破坏.

Background Traditional Chinese medicine (TCM) typically attributes the etiopathogenesis of perimenopausal syndrome (PMS) to kidney deficiency in the TCM stratification system for diagnosis. However, the molecular basis of this classical attribution remains to be investigated. Aim of the Study. By unraveling the responses to TCM treatment for kidney deficiency, the metabolomic link between PMS and kidney deficiency can be evaluated for in-depth understanding of the mechanism of TCM treatment and development of better treatment protocols. Materials and Methods With naturally aged rats as a model for PMS, the metabolomic response to TCM treatment for kidney deficiency was investigated by 1H NMR. Results 1H NMR metabolomic evidence of plasma samples demonstrates that treatments with two classical TCM prescriptions for kidney deficiency, decoctions of Yougui and Zuogui, result in modulating the metabolic state of the disease model towards that of rats of younger age. Conclusion The data support the notion that kidney deficiency is responsible, in part at least, for PMS, and the relevant prescriptions are helpful in dampening the changes in the body's metabolic states to alleviate symptoms of the disorder.

目的:观察益气化瘀方联合过表达Mash-1修饰的胚胎干细胞对小鼠脊髓损伤的修复作用.方法:采用钳夹法建立小鼠急性脊髓损伤模型,于造模后第3天将Mash-1修饰的胚胎干细胞移植到脊髓损伤部位,并给予益气化瘀方干预.通过观察小鼠的后肢功能评分、脊髓的剩余面积以及神经营养因子、炎症因子和凋亡相关基因的表达改变;并采用免疫荧光检测OCT3/4、nestin、β-tubulinⅢ、GFAP的阳性细胞数目,分析移植细胞在体内的存活和分化情况.结果:与模型组比较,益气化瘀方可减少炎症因子TNF-α和凋亡基因caspase-3的表达,轻微提高小鼠运动功能.与单纯干细胞移植组比较,益气化瘀方联合Mash-1修饰胚胎干细胞移植能减少炎症因子TNF-α的基因表达水平,提高移植细胞的存活数量,进而改善小鼠的后肢运动评分.结论:益气化瘀方能减轻脊髓损伤的炎症反应,增加移植细胞在脊髓损伤部位的存活数量,促进脊髓损伤的修复.

Cervical spondylotic myelopathy (CSM) is a leading cause of spinal cord dysfunction with few treatment options. Although mitochondrial dynamics are linked to a wide range of pathological changes in neurodegenerative diseases, a connection between aberrant mitochondrial dynamics and CSM remains to be illuminated. In addition, mechanisms underlying the emerging anti-inflammatory and neuroprotective effects of echinacoside (ECH), the main active ingredient of Cistanche salsa, are poorly understood. We hypothesized that excessive mitochondrial fission plays a critical role in regulating inflammatory responses in CSM, and ECH might alleviate such responses by regulating mitochondrial dynamics. To this end, we assessed the effects of ECH and Mdivi-1, a selective inhibitor of dynamin-related protein (Drp1), in a rat model of chronic cervical cord compression and activated BV2 cells. Our results showed that rats with Mdivi-1 intervention had improved motor function compared with vehicle-treated rats. Indeed, Mdivi-1 treatment attenuated pro-inflammatory cytokine expression, as well as activation of the nod-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, nuclear transcription factor-κB (NF-κB), and Drp1 in lesions. Compared with vehicle-treated rats, compression sites of Mdivi-1-treated animals exhibited elongated mitochondrial morphologies and reduced reactive oxygen species (ROS). Similarly, ECH-treated rats exhibited neurological recovery and suppression of inflammatory response or related signals in the lesion area after treatment. Interestingly, ECH treatment partly reversed aberrant mitochondrial fragmentation and oxidative stress within the lesion area. In vitro data suggested that ECH suppressed activated microglia by modulating activation of the NLRP3 inflammasome and NF-κB signaling. Furthermore, we observed that ECH markedly inhibited Drp1 translocation onto mitochondria, whereby it regulated mitochondrial dynamics and ROS production, which act as regulators of NLRP3 inflammasome activation and NF-κB signaling. Thus, our findings reveal that mitochondrial dynamics modulate inflammatory responses during CSM. Moreover, ECH may attenuate neuroinflammation in rats subjected to chronic cervical cord compression by regulating Drp1-dependent mitochondrial fission and activation of downstream signaling.

颈椎病的发病率逐年增高,而神经根型颈椎病是较常见的一种分型,其特点是出现与受压神经根分布区域相符合的疼痛、麻痹等不适症状,对患者的日常生活、工作、学习均造成较大的影响.从针灸、牵引、理疗、手法、封闭、药物等方法对神经根型颈椎病的治疗现状进行综述.

Astragalus membranaceus (AM) is a commonly used herb in traditional Chinese medicine (TCM), which has been used as an essential tonic to treat various diseases for more than 2000 years. In this study, we aimed to investigate the biological effects of extract from AM on breast cancer cell and its mechanism.

There have been various reports in the literature of an in vivo model for giant cell tumor of bone (GCTB). However, few suitable animal models of GCTB have been established, due to the fact that GCTB contains three histologically different cell types. To the best of our knowledge, injection of patient-derived GCTB cells into bone environment has not been reported until now. In the present study, the biological behavior of GCTB cells in nude mice was investigated through intratibial injection of patient-derived GCTB cells. Patient-derived GCTB cells were obtained from 5 patients who had not undergone chemo- and radiotherapy. Once isolated, the cell suspension was injected into the tibias of nude mice. The growth process was monitored by weekly observation and photographic documentation using X-ray. Four months after injection, nude mice were sacrificed and the injected tibial samples were fixed, and further analyzed using micro-computed tomography (micro-CT), standard histology, tartrate-resistant acid phosphatase (TRAP) staining and mitochondrial immunofluorescence staining. X-ray, micro-CT and standard histology revealed osteolytic destruction in the proximal end of the tibia. TRAP staining identified TRAP-positive, osteoclast-like cells distributed in the bone marrow interface of the lesion area. Anti-human mitochondrial immunofluorescence staining confirmed that the surviving cells in the osteolytic destruction were of human GCTB cell origin. These findings indicate that intratibial injection of patient-derived GCTB cells may elicit osteolytic destruction in nude mice. The results of the current study present a novel animal model for GCTB, opening new perspectives to investigate this disease and develop therapeutic agents.

LGR4 has been identified as a new receptor for RANKL in bone cells where it opposes RANK signaling to inhibit osteoclasts differentiation, and its therapeutic targeting promotes reduced bone loss in three mouse models of osteoporosis.

目的 观察芒柄花素对不同亚型乳腺癌细胞周期相关基因和蛋白表达的影响.方法 乳腺癌细胞MCF-7、SK-BR-3、MDA-MB-231经0、40、80 μmol/L芒柄花素作用48 h后,分别采用Trizol和RIPA裂解细胞,收集细胞的总RNA和总蛋白,进一步采用实时定量PCR技术和Western blotting技术检测细胞周期调控因子cyclin D1、cyclin E、p21、p27的基因和蛋白表达情况.结果 3种乳腺癌细胞cyclin D1和cyclin E的基因及蛋白表达水平随着药物浓度的增加而降低,而p21和p27的基因和蛋白表达水平随药物浓度的增加而升高,与对照组比较,差异显著(P＜0.05、0.01).结论 芒柄花素可以通过调控cyclin Dl、cyclin E、p21和p27基因和蛋白的表达,将乳腺癌细胞阻滞于G1期,从而抑制乳腺癌细胞的增殖.

Little information has been published in the literature regarding survival outcomes of patients with Ewing’s sarcoma family tumors (ESFTs) of the spine. The purpose of this study is to explore factors that may affect the prognosis of patients with non-metastatic spinal ESFTs. A retrospective analysis of survival outcomes was performed in patients with non-metastatic spinal ESFTs. Univariate and multivariate analyses were employed to identify prognostic factors for recurrence and survival. Recurrence-free survival (RFS) and overall survival (OS) were defined as the date of surgery to the date of local relapse and death. Kaplan–Meier methods were applied to estimate RFS and OS. Log-rank test was used to analyze single factors for RFS and OS. Factors with p values ≤0.1 were subjected to multivariate analysis. A total of 63 patients with non-metastatic spinal ESFTs were included in this study. The mean follow-up period was 35.1 months (range 1–155). Postoperative recurrence was detected in 25 patients, and distant metastasis and death occurred in 22 and 36 patients respectively. The result of multivariate analysis suggested that age older than 25 years and neoadjuvant chemotherapy were favorable independent prognostic factors for RFS and OS. In addition, total en-bloc resection, postoperative chemotherapy, radiotherapy and non-distant metastasis were favorable independent prognostic factors for OS. Age older than 25 years and neoadjuvant chemotherapy are favorable prognostic factors for both RFS and OS. In addition, total en-bloc resection, postoperative chemotherapy, radiotherapy and non-distant metastasis are closely associated with favorable survival.

Objective To investigate the effects of overexpression of Mash-1 gene on functional recovery and neural differentiation of embryonic stem cells in spinal cord injury mice. Methods CE3 cell line with overexpression of Mash-1 gene was generated with murine stem cell virus. Spinal cord injury model was established with forceps compression in 4-week-old KM mice. Normal saline (model group, n=12), CE3 cells with or without overexpression Mash-1 gene (CE3-Mash-1 and CE3 groups, n=12 respectively) were transplanted into the ar-eas of injury 3 days after injury. They were assessed with the Basso Mouse Scale (BMS) 1, 7, 14, 21, and 28 days after injury. 6 mice from each group were sacrificed 14 and 28 days after injury respectively. The spinal cord area remained were observed with HE stained, and the expression of Oct3/4, nestin,β-tubulin III and glial fibrillary acidic protein (GFAP) were detected with immunofluorescence in the injured spinal cord in the CE3 and CE3-Mash-1 groups. Results The score of BMS significantly improved in CE3 and CE3-Mash-1 groups com-pared with that of the model group (F>84.471, P<0.05), with the more area of spinal cord remained (F>49.990, P<0.05). There were less Oct3/4 positive cells in the CE3-Mash-1 group than CE3 group (t=5.439, P<0.001), with more nestin (t=-7.536, P<0.001) andβ-tubulin III (t=-9.941, P<0.001) positive cells. However, there was no significant difference in GFAP positive cells between CE3-Mash-1 and CE3 groups (t=1.701, P>0.05). Conclusion Overexpression of Mash-1 gene promotes CE3 cells to differentiate into neurons in spinal cord injury mice, and improve the motor function recovery.

CEP hypertrophy is one of the characteristics of intervertebral disc degeneration (IDD). LIG exerts a protective effect on IDD in animal model. The effect of LIG on CEP hypertrophy is further investigated in the present study. Cells were isolated from hypertrophic samples obtained from patients during vertebral fusion surgery. Cellular proliferation and the expression of type I collagen (Col I) and TGF-β1 were tested. In the bipedal rats, the edges of the CEP and the sizes of noncartilaginous outgrowth, as well as the expression of osteogenic markers, Col1a, ALP, Runx2, and TGF-β1, were detected. Within two passages, the condensed hypertrophic CEP cells exhibited osteogenic capacity by bony-like nodules and ALP positive staining, along with increased expression of Col I and TGF-β1. LIG inhibited proliferation of CEP cells and downregulated the expression of Col I and TGF-β1 in vitro. Furthermore, LIG attenuated CEP hypertrophy on the lumbar spine of bipedal rats by reducing Col1a, ALP, Runx2, and TGF-β1 mRNA expression and TGF-β1 distribution in vivo. We concluded LIG exerted a preventive effect on CEP hypertrophy via suppression of TGF-β1 levels. This information could be used to develop alternative therapeutic methods to treat spinal CEP hypertrophy.

Giant cell tumor of bone(GCTB) is a special bone tumor for it consists of various cell types, and its biological characteristics is different from common benign or malignant neoplasm. In the present study, we report the biological features of a primary Asian GCTB cell line named GCTB28. We analyzed extensive properties of the GCTB28 cells including morphological observations, growth, cell cycle, karyotype, proliferation, proteins expression, surface biomarker verification, and tumorigenicity in nude mice. We found that the stromal cells of GCTB were endowed with self-renewal capacity and played dominant roles in GCTB development. Moreover, we confirmed that GCTB cells can be CD33(-)CD14(-) phenotype which was not in accord with previous study. This study provides an in vitro model system to investigate pathogenic mechanisms and molecular characteristics of GCTB and also provides a useful tool for researching the therapeutic targeting of GCTB.

Objective: To observe clinical effects of triple therapy on LDH. Methods: 220 cases of lumbar disc herniation were randomly divided into the control group and treatment group, 110 cases in each group. The control group was treated by traction and massage, and the treatment group was treated by warming acupuncture more, for 6 weeks. Results: In the treatment group, 70 cases&nbsp;were cured; 28 cases were effective; 12 cases were a little effective; the total effciency was 100%. In the control group, 12 cases were cured; 32 cases were markedly effective; 60 cases were effective; 6 cases were ineffective; the total efficiency was 94.5%. The total effciency in the treatment group was signiifcantly higher (P<0.01). Conclusion: Traction, massage and acupuncture was effective on LDH.

Abstract Giant cell tumor of bone (GCTB) is a rare and highly osteolytic bone tumor that usually leads to an extensive bone lesion. The purpose of this study was to discover novel therapeutic targets and identify potential agents for treating GCTB. After screening the serum cytokine profiles in 52 GCTB patients and 10 normal individuals using the ELISA assay, we found that NF-κB signaling–related cytokines, including TNFα, MCP-1, IL1α, and IL17A, were significantly increased in GCTB patients. The results were confirmed by IHC that the expression and activity of p65 were significantly increased in GCTB patients. Moreover, all of the NF-κB inhibitors tested suppressed GCTB cell growth, and bortezomib (Velcade), a well-known proteasome inhibitor, was the most potent inhibitor in blocking GCTB cells growth. Our results showed that bortezomib not only induced GCTB neoplastic stromal cell (NSC) apoptosis, but also suppressed GCTB NSC–induced giant cell differentiation, formation, and resorption. Moreover, bortezomib specifically suppressed GCTB NSC–induced preosteoclast recruitment. Furthermore, bortezomib ameliorated GCTB cell–induced bone destruction in vivo. As a result, bortezomib suppressed NF-κB–regulated gene expression in GCTB NSC apoptosis, monocyte migration, angiogenesis, and osteoclastogenesis. Particularly, the inhibitory effects of bortezomib were much better than zoledronic acid, a drug currently used in treating GCTB, in our in vitro experimental paradigms. Together, our results demonstrated that NF-κB signaling pathway is highly activated in GCTB, and bortezomib could suppress GCTB and osteolysis in vivo and in vitro, indicating that bortezomib is a potential agent in the treatment of GCTB. Mol Cancer Ther; 15(5); 854–65. ©2016 AACR.