BACKGROUND:With the aging of the global population,the incidence rate of osteoporosis is also increasing.It is very important to further understand its pathogenesis and propose new therapeutic targets.Recent studies have shown that ferroptosis is closely related to the pathogenesis of some bone diseases,such as inflammatory arthritis,osteoporosis and osteoarthritis. OBJECTIVE:To summarize the previous studies on the mechanism of ferroptosis in osteoporosis,so as to provide new therapeutic ideas and potential therapeutic targets for osteoporosis. METHODS:The first author used the computer to search the documents published from 2000 to 2022 in CNKI,WanFang,VIP,PubMed and Web of Science with the key words of"ferroptosis,osteoporosis,osteoblasts,osteoclasts,iron chelators,reactive oxygen species,nuclear factor erythroid 2-related factor 2,heme oxygenase-1,glutathione peroxidase 4,review"in Chinese and English.A total of 70 articles were finally included according to the inclusion criteria. RESULTS AND CONCLUSION:Ferroptosis is significantly different from necrosis,apoptosis and autophagy.In terms of cell morphology and function,it does not have the morphological characteristics of typical necrosis,nor does it have the characteristics of traditional apoptosis,such as cell contraction,chromatin condensation,the formation of apoptotic bodies and the disintegration of cytoskeleton.Contrary to autophagy,ferroptosis does not form a classical closed bilayer membrane structure(autophagic vacuole).Morphologically,ferroptosis is mainly manifested by obvious contraction of mitochondria,increased membrane density,and reduction or disappearance of mitochondrial cristae,which are different from other cell death modes.Iron overload can destroy bone homeostasis by significantly inhibiting osteogenic differentiation and stimulating osteoclast formation,leading to osteoporosis.Iron overload interferes with the differentiation of stem cells to osteoblasts,leading to a weakened osteoblast function and further imbalance of bone metabolism in the body,which eventually leads to osteoporosis.Stimulated by iron overload,osteoclast bone resorption is enhanced and bone loss exceeds new bone formation.Iron chelators have been proved to have osteoprotective effects by inhibiting osteoclast activity and stimulating osteogenic differentiation of osteoblasts.Its potential mechanism is related to inhibiting osteoclast differentiation and promoting osteoblast differentiation.Antioxidants can prevent reactive oxygen species production and inhibit bone absorption,thus improving bone metabolism and effectively preventing osteoporosis.

Objective To study the factors influencing the blood concentration of caspofungin(CPFG),construct a prediction model,and validate the predictive effect of the model,so as to provide reference for the individualised dosing of patients with fungal infections in haematology.Methods Seventy-five patients admitted to the Department of Haematology,General Hospital of Tianjin Medical University,who were treated with CPFG for antifungal therapy during the period of March 2021 to June 2022 were selected as the study subjects,and CPFG blood concentration monitoring was carried out to explore the influencing factors of CPFG blood concentration and to construct a prediction model accordingly.Hosmer-Lemeshow(H-L)was used to test the goodness-of-fit of the model,and another 30 patients were selected as the verification group,and the predictive effect of the model was verified by the receiver's operating characteristics(ROC)curve.Results The mean blood concentrations of the patients at 0.5,9 and 24 h were(12.54±4.38),(6.80±2.76),(4.13±2.16)μg·mL-1,and the mean AUC0-24h were(152.05±57.60)μg·mL-1·h.AUC0-24h was lower than the reference value(98 μg·mL-1·h)in two patients.The results of correlation analysis showed that gender showed a correlation with 0.5 h blood concentration(P＜0.05),and there was no correlation with the rest of the two time points blood concentration and AUC0-24h(P＞0.05).Body weight and albumin(Alb)concentration showed correlation with 0.5,9,24 h blood drug concentration and AUC0-24 h(P＜0.05),and the rest of the indicators showed no correlation with blood drug concentration and AUC0_24h at each time point(P＞0.05).The results of multifactorial analysis showed that the factors influencing the patients'0.5 h blood concentration were gender,Alb concentration and body weight,and the factors influencing the 9 and 24 h blood concentration and AUC0-24h were Alb concentration and body weight(P＜0.05).Correlation analysis showed that the daily dose was positively correlated with the plasma concentration of CPFG at 0.5,9 and 24 h and AUC0-24h(P＜0.05).The results of multivariate analysis showed that the daily dose was also one of the influencing factors of the plasma concentration of CPFG(P＜0.05).ROC curve shows that the model has good prediction ability.Conclusion Body weight and Alb are significantly associated with CPFG blood concentrations and area under the drug-time curve,which can be used as a basis for preventive risk avoidance.

Objective The leptin receptor,encoded by the LEPR gene,is involved in tumorigenesis.A potential functional variant of LEPR,rs1137101(Gln223Arg),has been extensively investigated for its contribution to the risk of digestive system(DS)cancers,but results remain conflicting rather than conclusive.Here,we performed a case-control study and subsequent meta-analysis to examine the association between rs1137101 and DS cancer risk. Methods A total of 1,727 patients with cancer(gastric/liver/colorectal:460/480/787)and 800 healthy controls were recruited.Genotyping of rs1137101 was conducted using a polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)assay and confirmed using Sanger sequencing.Twenty-four eligible studies were included in the meta-analysis. Results After Bonferroni correction,the case-control study revealed that rs1137101 was significantly associated with the risk of liver cancer in the Hubei Chinese population.The meta-analysis suggested that rs1137101 is significantly associated with the risk of overall DS,gastric,and liver cancer in the Chinese population. Conclusion The LEPR rs1137101 variant may be a genetic biomarker for susceptibility to DS cancers(especially liver and gastric cancer)in the Chinese population.

Objective To observe the value of ultrasound-guided intra-articular injection of hypertonic glucose(HG)for treating knee osteoarthritis in plateau area.Methods Ninety-one patients with knee osteoarthritis in plateau area(124 affected knee joints)were enrolled.The knee joints were divided into 20％HG group(n=67),25％HG group(n=42)or sodium hyaluronate group(n=15)based on the medication.Clinical and ultrasound scores were compared before and after treatment,and the efficacy of injection of HG was evaluated.Results At the 12th and 48th weeks after treatment,visual analog scale(VAS)scores decreased in all 3 groups(all P＜0.05),and Lysholm scores of 20％HG group and 25％HG group increased compared to those before treatment(all P＜0.05).The difference of Lysholm score before and in the 12th week after treatment,and of VAS score before and in the 48th week after treatment of 25％HG group were higher than those of 20％HG group(both P＜0.05).The joint exudation score of 20％HG group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial blood flow score decreased in the 12th and 48th weeks after treatment compared to those before treatment(both P＜0.05).The joint exudation score of 25％HG group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial hyperplasia and synovial blood flow score decreased in the 12th and 48th weeks after treatment compared to before treatment(all P＜0.05).The joint exudation score of sodium hyaluronate group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial hyperplasia score decreased in the 12th and 48th weeks after treatment compared to that before treatment(both P＜0.05).Conclusion Ultrasound-guided intra-articular injection of HG,esp.25％HG,had certain value for treating knee osteoarthritis in plateau area.

Objective To reveal the material base of traditional Chinese medicine(TCM)syndrome types of chronic aplastic anemia(CAA)by investigating the differences of fecal non-targeted metabolomics changes in patients with CAA and normal people,and by analyzing the differential fecal metabolite profiles in CAA patients with various TCM syndrome types.Methods The analysis was performed on 21 CAA patients(CAA group)who were initially treated at Affiliated Hospital of Nanjing University of Chinese Medicine from July 2018 to June 2021 and on 24 volunteers who had healthy medical checkup(normal group)during the same period.The 21 cases of CAA patients were categorized into four cases of kidney yang deficiency group,12 cases of kidney yin deficiency group,and five cases of deficiency of kidney yin and yang group according to the criteria of TCM syndrome differentiation and classification.Differential metabolites were screened after stool collection,gas chromatography-mass spectrometry(GC-MS)analysis and multivariate statistical analysis,and then enrichment analysis of the metabolic pathway was conducted.Results(1)Obvious differences were presented in 38 kinds of metabolites between the CAA group and the normal group,of which there were 27 metabolites,including 3-hydroxyphenylacetic acid,adenine,isocitric acid,L-glutamine,uric acid and guanine,having the up-regulated relative contents in CAA group,while there were 11 metabolites,including 3'-adenosine,eicosanoic acid,inosine,N-acetylornithine and norepinephrine,having the down-regulated relative contents in CAA group.The primary enriched metabolic pathways of the differential metabolites included arginine biosynthesis,purine metabolism,central carbon metabolism in cancer,glyoxylate and dicarboxylic acid metabolism,GABAergic synapses,metabolism of alanine,aspartate and glutamate,and D-glutamine and D-glutamate metabolism.(2)There were significant differences in 16 kinds of metabolites among the CAA patients with various TCM syndrome types,including 5-methoxytryptamine,epinephrine,arbutin,β-alanine,pentanediamine,inositol,pyruvate and tyramine,etc.The primary enriched metabolic pathways of the differential metabolites included neuroactive ligand-receptor interactions,tyrosine metabolism,phosphatidylinositol signaling system,pantothenic acid and coenzyme A biosynthesis,glycolysis/gluconeogenesis,primary bile acid biosynthesis,phosphoinositide metabolism,ascorbic acid and aldehyde acid metabolism,type Ⅱ diabetes mellitus,and adrenergic signal transduction in cardiomyocytes.Conclusion Significant differences exist in the fecal metabolomics between the CAA patients and the normal volunteers,which may reflect the metabolic changes of immune dysregulation and hematopoietic failure mediated by the altered intestinal flora in CAA patients.This study revealed the material base of CAA of various kidney deficiency syndromes for the first time at the level of fecal metabolomics,which provides a novel approach for the study of modernization of TCM syndrome.

Objective To evaluate the clinical efficacy of Ningxin decoction in treating patients with permanent atrial fibrillation(AF)of deficiency of both qi and yin type infected by coronavirus disease 2019(COVID-19).Methods From February to August 2023,120 patients with permanent AF of deficiency of both qi and yin type infected by COVID-19 were collected from the outpatient clinic of Guang'anmen Hospital South Campus,China Academy of Chinese Medical Sciences.Patients were randomly divided into a control group and a treatment group using a random number table method,there were 60 cases in each group,and 1 case fell off in each group,leaving 59 cases in each group.The treatment group was given Ningxin decoction(drug composition:Cassia branch 15 g,Cornus officinalis 15 g,Rehmannia 15 g,Rhodiola rosea 15 g,Ginseng 6 g,Mulberry parasitica 10 g,Coptis coptis 3 g,Salvia miltiorrhiza 12 g,Ligusticum chuanxiong 10 g,Ophiopogon 12 g,and schisandra Vinegar 6 g)using formula granules to rinse with water,twice a day.The control group was given 10 mg of coenzyme Q10 twice a day,and both groups were treated continuously for 4 weeks.The improvements in traditional Chinese medicine(TCM)symptoms,including chest pain,dizziness,fatigue,lethargy,palpitations,chest tightness and shortness of breath,insomnia,and excessive dreams before and after treatment in two groups,as well as changes in heart rate variability(HRV)and heart rate deceleration indicators were observed.Results The treatment group showed significant therapeutic effects in improving chest pain,fatigue,palpitations,chest tightness and shortness of breath,there were also improvements in lethargy,insomnia,and excessive dreaming,there was no significant therapeutic effect in improving dizziness.The control group showed significant therapeutic effects in improving palpitations,there were also improvements in chest pain,chest tightness and shortness of breath,while there was no significant therapeutic effect in other aspects.The treatment group showed significantly higher effective rates in improving chest pain,fatigue,lethargy,chest tightness and shortness of breath,insomnia,and excessive dreaming compared to the control group[chest pain:95.45%(42/44)vs.78.57%(33/42),fatigue:83.67%(41/49)vs.31.25%(15/48),lethargy:73.91%(34/46)vs.12.77%(6/47),chest tightness and shortness of breath:88.89%(48/54)vs.63.64%(35/55),insomnia:78.43%(40/51)vs.12.24%(6/49),excessive dreaming:76.09%(35/46)vs.43.75%(21/48),all P<0.05].The standard deviation of all normal to normal RR intervals(SDNN),root mean square of the difference between adjacent NN intervals(RMSSD),standard deviation of all 5-minute RR intervals(SDANN),and percent of NN50 in the total number of NN intervals(PNN50)in both groups after treatment were significantly increased compared to before treatment,and the RMSSD,SDANN,and PNN50 in the treatment group were significantly higher than those in the control group after treatment[RMSSD(ms):28.96±3.59 vs.24.34±2.66,SDANN(ms):108.55±11.80 vs.100.44±13.58,PNN50:7%(6%,7%)vs.5%(5%,6%),all P<0.05].There was no statistically significant difference in heart rate deceleration between the two groups before and after treatment.Conclusion Ningxin decoction has a good effect on the improvement of TCM syndromes and heart rate variability in patients with permanent atrial fibrillation of deficiency of both qi and yin type after being infected by COVID-19.

Objective:To investigate the effect of recombinant lipoproteins of Brucella outer membrane protein 16, 19 (L16 and L19) on the expression of immune regulatory factors in human monocytic leukemia cell line (THP-1 cells). Methods:THP-1 cells activated with phorbol ester (PMA) were used as an in vitro experimental cell model, and a group design was used to co-culture L16, L19 and THP-1 cells (L16 stimulated group, L19 stimulated group), respectively. THP-1 cells activated with PMA were used as the control group. When co-cultured for 4 hours, immunofluorescence staining (IFS) and Western blotting were used to detect whether L16 and L19 entered the cells, respectively; when co-cultured for 12, 24 hours, real-time fluorescent quantitative PCR was used to measure the mRNA expression levels of interferon regulatory factor 1 (IRF-1) and trans activator protein of major histocompatibility complex class Ⅱ (CⅡTA); Western blotting was used to detect the protein expression levels of T cell immunoglobulin mucin-3 (Tim-3) and γ interferon receptor 1 (IFNGR1). Results:When co-cultured for 4 hours, L16 and L19 were observed entering THP-1 cells in the L16 stimulated group and L19 stimulated group, respectively. When co-cultured for 12 hours, the expression level of IRF-1 mRNA in the L16 stimulated group (0.16 ± 0.15) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 24 hours, the expression level of CⅡTA mRNA in the L16 stimulated group (0.17 ± 0.10) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 12 and 24 hours, there were no statistically significant differences in the expression levels of IRF-1 and CⅡTA mRNA between the L19 stimulated group and the control group ( P > 0.05). Western blotting results showed that there were statistically significant differences in the expression levels of INFGR1 and Tim-3 protein among the control group, L16 stimulated group, and L19 stimulated group after co-cultured for 12 and 24 hours ( F = 50.92, 6.80, 148.73, 156.57, P < 0.05). Among them, when co-cultured for 12 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were significantly lower than that in the control group, and the L19 stimulated group was higher than the L16 stimulated group ( P < 0.05), and the expression level of Tim-3 protein in the L19 stimulation group was higher than that in the control group ( P < 0.05). When co-cultured for 24 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were lower than that in the control group, and the L19 stimulated group was higher than that in the L16 stimulated group ( P < 0.05); and the expression level of Tim-3 protein in the L16 stimulated group was higher than that in the control group and L19 stimulated group ( P < 0.05). Conclusions:Brucella L16 can downregulate the expression levels of IRF-1 and CⅡTA mRNA in THP-1 cells. Both L16 and L19 can downregulate IFNGR1 and upregulate Tim-3 protein expression levels.

This manuscript aims to investigate the effects of resibufogenin on the proliferation, migration and invasion of human hepatocellular carcinoma cells and its related mechanisms. MTT assay was used to determine the inhibitory effect of resibufogenin on the growth of four hepatocellular carcinoma cells in vitro. Wound-healing assay and Transwell assay were used to evaluate the migration and invasion ability of resibufogenin on MHCC-97H cells. Western blot assay was used to detect the expression of migration and invasion related proteins in MHCC-97H cells treated with different concentrations of resibufogenin. The results showed that resibufogenin significantly inhibited the proliferation of hepatocellular carcinoma cells in vitro. The half maximal inhibitory concentration (IC₅₀) values on MHCC-97H, HepG2, SK-Hep-1 and Huh-7 cells were 0.55 ± 0.06, 2.83 ± 0.24, 5.25 ± 0.49, 14.89 ± 2.28 μmol·L^-1, respectively. Resibufogenin also suppressed the migration and invasion of MHCC-97H cells in a concentration-dependent manner. The protein expression of integrin α2, integrin α6, integrin β1, N-cadherin, matrix metalloproteinase 2 (MMP2) and transcription factor Twist in MHCC-97H cells were decreased significantly with the increase of the concentration of resibufogenin, while the protein expression of E-cadherin increased. In addition, we found that p-PI3K/PI3K and p-AKT/AKT ratios were significantly reduced after treatment with resibufogenin. In conclusion, resibufogenin can inhibit the proliferation, migration and invasion of hepatocellular carcinoma MHCC-97H cells in vitro, which is related to the regulation of intracellular migration and invasion protein expression and PI3K/AKT signaling pathway.

Tanshinone IIa is a key ingredient extracted from the traditional Chinese medicine Salvia miltiorrhiza (Danshen), and is widely used to treat various cardiovascular diseases. Vascular calcification is a common pathological change of cardiovascular tissues in patients with chronic kidney disease, diabetes, hypertension and atherosclerosis. However, whether Tanshinone IIa inhibits vascular calcification and the underlying mechanisms remain largely unknown. This study aims to investigate whether Tanshinone IIa can inhibit vascular calcification using high phosphate-induced vascular smooth muscle cell and aortic ring calcification model, and high dose vitamin D₃ (vD₃)-induced mouse models of vascular calcification. Alizarin red staining and calcium quantitative assay showed that Tanshinone IIa significantly inhibited high phosphate-induced vascular smooth muscle cell and aortic ring calcification. qPCR and Western blot showed that Tanshinone IIa attenuated the osteogenic transition of vascular smooth muscle cells. In addition, Tanshinone IIa also significantly inhibited high dose vD₃-induced mouse aortic calcification and aortic osteogenic transition. Mechanistically, Tanshinone IIa inhibited the activation of NF-κB and β-catenin signaling in normal vascular smooth muscle cells. Similar to Tanshinone IIa, inhibition of NF-κB and β-catenin signaling using the chemical inhibitors SC75741 and LF3 attenuated high phosphate-induced vascular smooth muscle cell calcification. These results suggest that Tanshinone IIa attenuates vascular calcification at least in part through inhibition of NF-κB and β-catenin signaling, and Tanshinone IIa may be a potential drug for the treatment of vascular calcification.
Animals ; Mice ; NF-kappa B/metabolism* ; beta Catenin/metabolism* ; Signal Transduction ; Myocytes, Smooth Muscle/metabolism* ; Vascular Calcification/metabolism* ; Phosphates/metabolism*

Aim To reveal the aetion mechanism of Trillium tschonoskii Maxim (TTM) in the treatment of myoeardial ischemia ( MI) by using network pharma¬cology combined with molecular docking.Methods Compounds of TTM were detected and fished out from TCMID, TCM@TAIWAN , BATMAN-TCM database, and the literature data from PubMed , CNK1, and WAN- FANGD database.PharmMapper database was used to find the targets related to compounds, and DISGeNET, GeneCards, DrugBank and OMIM databases were used to find the targets related to Ml.The predictive targets of TTM in the treatment of Ml were obtained.Cytosca- ope 3.1.2 Software and String database were used to build compound-target network and PP1 network.Gene ontology ( GO ) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes ( KEGG ) pathway enrichment analysis were performed by utili¬zing the CludterProfiler Software package of RStudio software.The molecular docking was used for verifying the results of network analysis.Results The 10 active compounds of TTM were screened , and 13 core targets of Ml were predicted, such as ALB, EGFR, MAPK1 , CASP3,ESR1 ,etc.A total of 28 Ml-related signaling pathways were fished out.The results of molecular docking showed that the core active ingredients had good binding activity with the key targets.Conclusion TTM may play a role in the treatment of Ml through regulating multiple ingredients, multiple pathways, and multiple targets.