BACKGROUND:Accumulating evidence shows that Panax notoginseng, a well-known medicinal herb, has an ideal effect on prevention and treatment of skeletal diseases. In this study, we reviewed clinical applications of clinical application as well as phytochemistry, pharmacokinetics, pharmacology in improving bone quality and toxicity of Panax notoginseng. PURPOSE:Review the phytochemistry, pharmacokinetics, pharmacology involved in the improving bone metabolism and toxicity of Panax notoginseng and evaluate its potential as a traditional Chinese herbal medicine for osteoporosis. METHODS:Several databases were consulted, including PubMed, China National Knowledge Infrastructure, National Science and Technology Library and Web of Science. The following words or phrases were used alone or in combinations in the titles and/or abstracts: "","Panax notoginseng", "Sanqi", "osteoporosis", "bone", "osteoblast", "osteoclast", "phytochemistry", "pharmacology" and "pharmacokinetics". Altogether 160 papers were cited. RESULTS:8 clinical trials of Panax notoginseng alone for the treatment of osteoporosis were identified, most of which used traditional Chinese patent medicines to treat osteoporosis fractures. In these clinical trials, Panax notoginseng preparations have achieved relatively good therapeutic effects. However, more rigorous large-scale experiments are expected to prove their efficacy. Phytochemistry study showed that saponins, flavonoids, polysaccharides are the main active ingredients extracted from Panax notoginseng and the transformation of saponins during the processing explains the different effects of raw and cooked Panax notoginseng. The pharmacokinetics data reveals that protopanaxdiol-type (ppd-type) saponins possesses higher bioavailability than protopanaxtriol-type(ppt-type) saponins and ppd-type saponins such as ginsenoside Ra3, Rb1, and Rd can represent suitable pharmacokinetic markers for Panax notoginseng extracts. The data from animal experiments demonstrates that Panax notoginseng can improve bone quality in ovariectomized, diabetic, hyperlipidemia, radiation-induced, and arthritis rats through the regulation of anti-adipogenesis, anti-inflammation, anti-oxidation, angiogenesis and estrogenic effects. In vitro experiments, the activities of improving bone quality of Panax notoginseng and its ingredients may be attributed to the regulation of multiple signaling pathways, including Wnt/β-catenin, BMP/BMP-R, AMPK/mTOR, GPER/PI3K/AKT, etc. Acute and chronic toxicity as well as genotoxicity studies show that Panax notoginseng is well tolerated while long term use may lead to liver and kidney toxicity. CONCLUSIONS:Panax notoginseng is a superior medicinal herb that contains multiple active ingredients and could play a potential role in the prevention and treatment of osteoporosis. Further studies should concentrate on developing Panax notoginseng products with higher curative effect and bioavailability.

Cadmium (Cd), a heavy metal pollutant that accumulates in organisms, is osteotoxic and contributing to the development of osteoporosis. Lycium barbarum polysaccharide (LBP), a polysaccharide obtained from the Chinese medicinal plant Lycium barbarum, exhibits protective effects against oxidative stress and cellular damage. However, it is unclear whether LBP can effectively inhibit Cd-induced osteotoxicity. This study investigated whether LBP alleviates Cd-induced osteogenic suppression in bone marrow mesenchymal stem cells (BMSCs) via autophagy modulation. Cell viability, osteogenic markers (ALP, BMP2, COL1, RUNX2), and mineralization were assessed using CCK-8, qPCR, Western blot, and alizarin red staining. Autophagy flux was evaluated via GFP-mCherry-LC3B transfection and protein markers (LC3-II, P62, BECN). We confirmed that Cd inhibited the formation of cellular autophagosomes while adversely affecting BMSCs. However, LBP mitigated the Cd-induced osteogenic inhibitory effect by stimulating the autophagy processes and facilitating the formation of autophagosomes and autolysosomes. In conclusion, LBP is a potential bone-enhancing agent that can improve cell viability and osteogenic differentiation while mitigating the detrimental effects of Cd on BMSCs through the activation of autophagy.

ObjectivesThis study was aimed to develop a nomogram that can accurately predict the likelihood of cognitive dysfunction in individuals with abdominal obesity by utilizing various predictor factors.MethodsA total of 1490 cases of abdominal obesity were randomly selected from the National Health and Nutrition Examination Survey (NHANES) database for the years 2011–2014. The diagnostic criteria for abdominal obesity were as follows: waist size ≥ 102 cm for men and waist size ≥ 88 cm for women, and cognitive function was assessed by Consortium to Establish a Registry for Alzheimer’s Disease (CERAD), Word Learning subtest, Delayed Word Recall Test, Animal Fluency Test (AFT), and Digit Symbol Substitution Test (DSST). The cases were divided into two sets: a training set consisting of 1043 cases (70%) and a validation set consisting of 447 cases (30%). To create the model nomogram, multifactor logistic regression models were constructed based on the selected predictors identified through LASSO regression analysis. The model’s performance was assessed using several metrics, including the consistency index (C-index), the area under the receiver operating characteristic (ROC) curve (AUC), calibration curves, and decision curve analysis (DCA) to assess the clinical benefit of the model.ResultsThe multivariate logistic regression analysis revealed that age, sex, education level, 24-hour total fat intake, red blood cell folate concentration, depression, and moderate work activity were significant predictors of cognitive dysfunction in individuals with abdominal obesity (p < 0.05). These predictors were incorporated into the nomogram. The C-indices for the training and validation sets were 0.814 (95% CI: 0.875-0.842) and 0.805 (95% CI: 0.758-0.851), respectively. The corresponding AUC values were 0.814 (95% CI: 0.875-0.842) and 0.795 (95% CI: 0.753-0.847). The calibration curves demonstrated a satisfactory level of agreement between the nomogram model and the observed data. The DCA indicated that early intervention for at-risk populations would provide a net benefit, as indicated by the line graph.ConclusionAge, sex, education level, 24-hour total fat intake, red blood cell folate concentration, depression, and moderate work activity were identified as predictive factors for cognitive dysfunction in individuals with abdominal obesity. In conclusion, the nomogram model developed in this study can effectively predict the clinical risk of cognitive dysfunction in individuals with abdominal obesity.

ObjectiveTo optimize the automated radiosynthesis of the purinergic ion channel receptor 7 (P2X7R) imaging agent 18F-JNJ64413739 and evaluate its potential for brain imaging in osteoporotic model rats.MethodsA more electron-deficient nitropyridine was employed as the labeling precursor to facilitate the 18F-labeling. The radiosynthesis was conducted on an AllinOne synthesis module, and followed by purification via high-performance liquid chromatography (HPLC). The resulting 18F-JNJ64413739 was subjected to quality control tests. Small-animal PET/CT imaging studies were performed in sham and osteoporotic model rats.ResultsThe optimized automated radiossynthesis of 18F-JNJ64413739 was successfully completed in approximately 100 min with non-decay-corrected radiochemical yield of 6.7% ± 3.8% (n = 3), >97% radiochemical purity and >14.3 ± 1.3 GBq/μmol molar activity. The product met all clinical quality requirements. 18F-JNJ64413739 PET/CT imaging showed revealed significantly higher radioactivity uptake in various brain regions of the osteoporotic model rats compared to sham control group.ConclusionWe successfully optimized the automated radiosynthesis of 18F-JNJ64413739. The resulting tracer not only met clinical quality requirements but also demonstrated potential for clinical application in the diagnosis of osteoporosis, as evidenced by higher radioactivity uptake in various brain regions of osteoporotic model rats compared to normal controls.

ScopeGlycine is commonly used as an additive in bone health supplements, the activity and differentiation of bone mesenchymal stem cells (BMSCs) are essential to bone metabolism, but the effect of Glycine on bone metabolism and specific mechanism are not fully clarified.Methods and resultsThe ovariectomized rats to evaluate the effects of Glycine on bone quality and quantity is constructed; then used an ER signaling inhibitor (ICI182780) and an ERα deficient BMSCs to explore how Glycine mediated ERα regulating the osteogenic and adipogenic differentiation of BMSCs; furthermore, an autodock analysis is used to assess the affinity of Glycine and ERα. The results show that Glycine significantly moderated bone mass and bone microstructure in ovariectomized rats; Glycine stimulates the osteogenic differentiation and attenuates the adipogenic differentiation in OVX rats and BMSCs, and these effects could be abolished by ICI 182780; further docking experiment showes that Glycine and ERα have a stronger affinity, and finally proves that the impact of Glycine could be blocked by ERα.ConclusionGlycine stimulates osteogenesis and attenuates adipogenesis in ovariectomized rats, which process may involve in ERα mediated ER signaling pathway.

Excessive salt intake can induce a variety of diseases, such as hypertension, cardiovascular disease, kidney disease and so on,it is also one of the factors promoting bone resorption. The mechanism of osteoporosis -induced exacerbations of high salt diet is not well-defined. In this study, we used ovariectomized 6-month-old Sprague Dawley rats to construct a high bone turnover model, and then administrated with high sodium chlo-ride diet (2.0% w/w NaCl, 8.0% w/w NaCl) for 12 weeks to observe the effect of high salt diet on bone meta-bolism. The results showed that high salt diet could lead to the destruction of bone microstructure, promote the excretion of urinary calcium and phosphorus and accelerate the bone turnover, as well as cause the pathologic structural abnormalities in renal tubular. At the same time, it was accompanied by the up-regulated expression of the epithelial sodium channel (ENaC alpha), voltage-gated chloride channels (ClC)-3 and the down-regulated expression of Na-Cl cotransporter (NCC), sodium calcium exchanger (NCX1) in femoral tissue and renal tu-bules. These findings confirm that high salt diet can destroy the microstructure of bone by increasing bone resorption and affect some ion channels of bone tissue and renal tubule in ovariectomized rats.

To investigate the association between blood routine (BRT) parameters and bone loss as well as the possible mechanism of this association with bone loss in the middle- aged and elderly patients. A total of nine hundred and ninety-eight subjects (the total) aged≥40 years in General Hospital of Southern Theater Command of People's Liberation Army from March 2015 to January 2018 were enrolled in a cross-sectional studyPatients were divided into two groups as "at least osteopenia group" (including osteopenia, osteoporosis, and severe osteoporosis) and "normal group" according to the diagnosis standard of OP. The above indicators were analyzed and compared between the two groups. Binary multivariate logistic regression analysis was carried out to explore the association between BRT parameters and risk of bone loss in the subjects Mediation Effect was conducted to test endocrine variables as mediators in the correlation of BRT parameters with the bone loss for the possible mechanisms. There were 669 cases in the at least osteopenia group and 329 in the normal group. RBC, Hb, HCT and MCHC were all positively correlated with BMD of lumbar spine(L1-4), left femoral neck and left femur, along with lymph positively correlated with BMD of left femoral neck and left femur, respectively. Eosinophils(Eo) was positively correlated with BMD of L1-L4 and RDW-SD together with RDW-CV were both negatively correlated with BMD of left femoral neck and left femur. By binary multivariate logistic regression, only Hb was associated with bone loss and the OR value was 0.478 as protective factor for BMD. Estradiol (E2) and testosterone (T) had partial mediating effect on the association of BRT parameters with the risk of bone loss. The mediating effect of E2 on the relationship between WBC and bone loss accounted for 46.35% of the total effect, and that of T on the relationship between RBC, Hb, HCT, MCHC and bone loss accounted for 27%, 18.3%, 31.1% and 26.8%, respectively. Bone loss in the middle-aged and elderly population is accompanied with the change of BRT parameters and the erythrocyte indices tend to be more obvious especially the Hb, which indicates anemia had a potential connection with bone loss. Sex hormone, especially T, is an important mediating variable in the association between blood routine parameters and bone loss.

Abstract Plastrum Testudinis (PT) is known as Traditional Chinese Medicine, which has commonly been used to treat and prevent bone metabolism for many years. However, the pharmacological mechanisms have not yet been fully clarified. In this study, we constructed a bilateral ovariectomy model to simulate postmenopausal osteoporosis (OP), then performed intragastric administration of different doses (160, 80, 40 mg/kg/day) of PT for 10 weeks. After treatment, we used dual-energy X-ray absorptiometry to evaluate bone mineral density, and micro-computed tomography and hematoxylin and eosin staining to analyze bone microstructure, immunochemistry, western blotting and quantitative polymerase chain reaction to detect the expression of osteogenic differentiation-related factors; and miRNA over-expression to evaluate the effect of miR-214 on the differentiation of bone mesenchymal stem cells (BMSCs) and related target genes. PT moderated bone mass and bone microstructure, alleviated body weight, and exhibited no estrogen-like effects; promoted the expression of osteogenic differentiation factors in the femur and lumbar vertebrae, as well as facilitated the expression of the Wnt signaling-related factors LRP5, Wnt3a, GSK-3β, and β-catenin. In addition, miR-214 inhibited osteogenic differentiation of BMSCs and targeted the Wnt signaling-related factors Wnt3a and β-catenin, while PT ameliorated these effects. This study indicated that PT may act as an antagonist of miR-214 to stimulate bone formation through β-catenin-mediated Wnt signaling.

目的:探讨补肾活血中药复方含药血清(NKABRS)通过钠氯同向转运体(NCC)和电压门控性氯离子通道5(CLC-5)对高盐诱导的体外大鼠肾小管上皮细胞NRK-52E损伤、凋亡和骨代谢相关因子表达的影响及作用机制.方法:取SPF级2月龄雌性SD大鼠30只,随机分为对照组与补肾活血中药复方组,分别灌胃给药后制备对照血清和补肾活血中药复方含药血清并作用于体外培养的NRK-52E细胞,CCK-8法检测NKABRS对细胞活力的影响.体外培养的NRK-52E细胞,分为对照组、NKABRS(3％)组、NaCl(0.25 mol/L)组和NaCl+NKABRS组,CCK-8法检测各组细胞活力,流式细胞仪检测细胞凋亡率,Western blot和RT-qPCR法检测细胞内骨桥蛋白(OPN)、NCC、CLC-5、骨形态发生蛋白7(BMP-7)和1α-羟化酶(CYP27B1)的表达,免疫荧光法检测细胞内CLC-5和NCC的蛋白含量及分布.通过RNA干扰技术敲减NRK-52E细胞NCC和CLC-5的表达,RT-qPCR检测敲减后各组细胞内BMP-7、1α-羟化酶和OPN的表达.结果:NaCl(0.25 mol/L)可抑制体外NRK-52E细胞的活力并增加其细胞凋亡率(P<0.01),同时增加细胞NCC和OPN表达,并减少CLC-5、BMP-7和1α-羟化酶表达(P<0.01);NKABRS可增加NaCl诱导的细胞活力并降低细胞凋亡率(P<0.01),降低NCC和OPN表达,并提高CLC-5、BMP-7和1α-羟化酶的表达(P<0.01).沉默体外培养的NRK-52E细胞内NCC基因可下调OPN表达并增加BMP-7和1α-羟化酶表达(P<0.01),NKABRS可进一步提高BMP-7和1α-羟化酶表达(P<0.05);沉默细胞内CLC-5基因可增加OPN表达并减少BMP-7和1α-羟化酶表达(P<0.01),NKABRS可抑制OPN表达并上调BMP-7和1α-羟化酶表达(P<0.01).结论:NKABRS可减轻高浓度NaCl引起的体外NRK-52E细胞的损伤和凋亡,其机制可能与减少NCC表达并增加CLC-5及细胞内骨代谢正向调节因子1α-羟化酶和BMP-7的表达有关.

The bone microenvironment is crucial for the growth and development of different types of osteocytes. Small extracellular vesicles (sEVs) secreted by bone mesenchymal stem cells are delivered to target cells where their contents regulate biological functions. Here, we evaluated the osteogenic effects and mechanism of sEVs derived from Plastrum testudinis -preconditioned bone mesenchymal stem cells (PT-sEV). The osteogenic effects of PT-sEV were evaluated by the differentiation of osteoblasts and the alternation of bone quality and quantity in ovariectomized rats. The specific mechanism was explored by high-throughput sequencing and verified by transfection with the corresponding miRNA mimic and inhibitor. RNA-sequence identified a unique enrichment of a set of miRNAs in PT-sEV compared with sEVs derived from untreated BMSCs. Overexpression or inhibition in vitro indicated that the osteogenic inducing potential of sEVs was mainly attributable to miR-330-5p, one of the most dramatically downregulated miRNAs in the PT-sEV fraction. Dual luciferase reporter assays showed that miR-330-5p negatively regulated osteogenesis by directly binding to the 3′ untranslated region of Tnc . Additional experiments showed that Tnc regulated Wnt/β-catenin signaling, and rescue experiment showed that miR-330-5p could restore β-catenin expression; additionally, animal experiments indicated that Wnt signaling was inactivated in the ovariectomized rats. These data demonstrated the regenerative potential of PT-sEV, which induced osteogenic differentiation of pre-osteoblasts, leading to bone formation. This process was achieved by delivering miR-330-5p, which regulated Tnc to control Wnt/β-catenin signaling.

A diet high in sodium chloride (NaCl) can affect renal function damage and increase urinary calcium excretion, leading to bone loss. in renal tubules, Na-Cl co-transporter (NCC) and chloride channel 5 (CLC-5) are involved in regulating urinary calcium excretion. In addition, some cytokines, such as Bone morphogenetic protein 7 (BMP-7) and 1α-hydroxylase, are synthesized by renal tubules, which target on bone and play important roles on bone metabolism. However, the specific mechanisms between NaCl and these ion channels or cytokines still need investigations from many aspects. This study, in culture normal rat renal tubular epithelial NRK-52E cells, showed that high concentrations of NaCl significantly inhibited the cell viability and increased the cell apoptosis. High concentration of NaCl reduce bone mineral density (BMD), as demonstrated by the significantly increased mRNA and protein levels of NCC and osteopontin (OPN), but decreased the levels of CLC-5, BMP-7, and 1α-hydroxylase. In addition, we found that ovariectomized (OVX) rats on a high-salt diet for 12 weeks had altered levels of these indices in the renal cortices. Moreover, the BMD in fourth and fifth lumbar vertebra (LV4 and 5) and femurs were significantly decreased and bone microstructure was destroyed of these rats. We also demonstrated that high concentration of NaCl enhanced the inhibition of these cytokines which is beneficial to increase BMD, induced by modulating ion channels NCC and CLC-5. In conclusion, our results indicate that high concentration of NaCl reduce BMD by regulating ion channels NCC and CLC-5.

Dent disease has multiple defects attributed to proximal tubule malfunction including low-molecular-weight proteinuria, aminoaciduria, phosphaturia, and glycosuria. To understand the changes in kidney function of the Clc5 chloride/proton exchanger gene knockout mouse model of Dent disease, we examined gene expression profiles from proximal S1 and S2 tubules of mouse kidneys. We found many changes in gene expression not known previously to be altered in this disease. Genes involved in lipid metabolism, organ development, and organismal physiological processes had the greatest number of significantly changed transcripts. In addition, genes of catalytic activity and transporter activity also had a great number of changed transcripts. Overall, 720 genes are expressed differentially in the proximal tubules of the Dent Clcn5 knockout mouse model compared with those of control wild-type mice. The fingerprint of these gene changes may help us to understand the phenotype of Dent disease.