Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

Objective:To investigate the relationship between preoperative plasma fibrin degradation products (FDP) level and clinicopathological features of patients with completely resected non-small cell lung cancer (NSCLC).Methods:A retrospective case series study was performed. The clinical data of 521 patients who were pathologically diagnosed with NSCLC in Beijing Friendship Hospital Affiliated to Capital Medical University from January 2016 to December 2017 were retrospectively analyzed. Among 521 cases, 406 cases were postoperatively pathologically confirmed as non-lymph node and non-distant metastasis (non-metastasis group) and 115 cases were postoperatively pathologically confirmed as lymph node or distant metastasis (metastasis group). The preoperative FDP level and clinicopathological characteristics as well as the clinicopathological characteristics of NSCLC patients with different FDP levels were compared between the two groups. The correlation between preoperative FDP level and TNM staging was analyzed by using Spearman correlation analysis.Results:Among 521 NSCLC patients, 266 cases were female, 255 cases were male; the age [ M( Q1, Q3)] was 59 years (54 years, 65 years); 441 cases were adenocarcinoma and 70 cases were squamous cell carcinoma. The preoperative median FDP level was 2.78 mg/L (2.35 mg/L, 3.13 mg/L) and 2.99 mg/L (2.56 mg/L, 4.16 mg/L), respectively of NSCLC patients in non-metastasis group and metastasis group, and the difference was statistically significant ( Z = 6.13, P < 0.001). The preoperative FDP level was 2.56 mg/L (2.35 mg/L, 3.20 mg/L) and 2.99 mg/L (2.56 mg/L, 3.20 mg/L), respectively in the early-stage NSCLC (stage Ⅰ-Ⅱ) and advanced NSCLC (stage Ⅲ-Ⅳ) patients, and the difference was statistically significant ( Z = 8.42, P < 0.001). Spearman correlation analysis showed that preoperative FDP level was positively correlated with tumor diameter ( r = 0.287, P < 0.001). There was a positive correlation between preoperative FDP level and the number of metastatic lymph nodes in 115 patients with lymph node metastasis ( r = 0.679, P < 0.001). According to the preoperative median FDP (2.78 mg/L), all patients were divided into FDP ≤2.78 mg/L group and FDP >2.78 mg/L, and there were statistically significant differences in age, metastasis, tumor staging, tumor diameter, the metastatic number of lymph node and histological types of NSCLC patients in both groups (all P < 0.05). Conclusions:The increase of preoperative plasma FDP level may be related to the tumor metastasis and clinical stage of NSCLC patients

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

Objective:To investigate the effects of lncRNA associated with microvascular invasion in hepatocellular carcinoma (lncRNA MVIH) on proliferation, invasion and tumorigenic ability of colorectal cancer SW620 cells.Methods:SW620 cells cultured in vitro were divided into empty vector group (transfected with pcDNA3.1 empty vector plasmid), MVIH overexpression group (transfected with pcDNA3.1-MVIH overexpression vector plasmid), NC siRNA group (transfected with interference sequence negative control NC siRNA) and MVIH siRNA group (transfected with MVIH siRNA interference sequence). The mRNA expression level of MVIH in SW620 cells in each group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Cell counting kit-8 (CCK-8), clone formation test and transwell chamber were used to detect the proliferation activity, clone formation ability and invasion ability of SW620 cells; The subcutaneous transplanted tumor of nude mice was constructed by subcutaneous injection of SW620 cells in each group 48 hours after transfection. The transplanted tumor volume was measured every 5 days to evaluate the tumor growth. On the 25th day, the nude mice in each group were killed and the tumor weight was measured. Results:The mRNA MVIH expression level, cell proliferation, clone formation rate and number of invasive cells of SW620 cells in the MVIH overexpression group were significantly higher than those in the empty vector group ( P<0.05); the MVIH mRNA expression level, cell proliferation , clone formation rate and number of invasive cells of SW620 cells in the MVIH-siRNA group were significantly lower than those in NC-siRNA group ( P<0.05). The results of subcutaneous transplantation tumor in nude mice showed that the growth rate of tumor in MVIH overexpression group was faster and the tumor weight was higher than those in the empty vector group ( P<0.05); the growth rate of tumor in MVIH overexpression group was slower and the tumor weight was lower than those in NC-siRNA group ( P<0.05). Conclusions:LncRNA MVIH can promote the proliferation, invasion and tumorigenesis of SW620 cells in colorectal cancer.

Neutrophilic granule protein (NGP) was previously reported as a granular protein of neutrophils in mouse, but the function has not been known clearly. We found the presence of the possible signal peptide in NGP and validated this protein is circulating in the bloodstream. In our findings, NGP is being modified post-translationally in Golgi apparatus and endoplasmic reticulum, which is a universal character of secretory molecules with a signal peptide. The secreted NGP protein could be detected both in vitro and in vivo. NGP has sequence similarity with an antimicrobial protein cathelicidin, and we observed the aspect of inflammation of NGP. Interestingly, NGP interacts with the complex of LPS and LPS binding protein (LBP). This interaction blocks the binding of the complex of LPS and LBP to TLR4 and the downstream inflammatory signals. Furthermore, the inhibitory function of NGP against the inflammatory effect of LPS could be observed in both in vitro and in vivo. With these findings, we report NGP is a novel secretory protein to mask LPS and inhibit its function.
Animals ; Carrier Proteins ; Cytokines ; Endoplasmic Reticulum ; Golgi Apparatus ; In Vitro Techniques ; Inflammation ; Lipopolysaccharides ; Masks ; Mice ; Neutrophils ; Protein Binding ; Protein Sorting Signals

Objective To investigate the relationship between Ezrin expression and subcellular localization of E?cadherin(E?cad),and explore the clinical significance of this relationship to pathological features such as lymph nodes metastasis in breast cancer. Methods Ninety four cases of breast cancer tissue samples with lymph node metastasis were collected. The expression of Ezrin and E?cad was detected by immunohistochemi?cal method. Results The positive rates of and E?cad and Ezrin were respectively 45.7%and 58.5%in 94 nodes positive breast cancer,containing membranal expression of E?cad(E?cadm)in 20 cases and cytoplasmic expression of E?cad(E?cadc)in 23 cases;the frequency of E?cadc positive staining was significantly higher in Ezrin(+)tissues than that in Ezrin(-)tissues(P=0.025);E?cad expression level was significantly lower in TNMⅡ?Ⅲstage cases(P=0.001),and Ezrin expression(P=0.036)and E?cadc(P=0.013)was significantly increased in bigger cases;com?pared with E?cad(+),Ezrin(-),E?cadm tissues,the number of lymph node metastasis in E?cad(-)(P=0.011),Ezrin(+)(P=0.002),E?cadc (P=0.020)tissues were increased significantly;in the order of E?cad(+)/Ezrin(-),E?cad(-)/Ezrin(-),E?cad(+)/Ezrin(+),and E?cad(-)/Ezrin(+),the number of lymph node metastasis was increased significantly(P<0.001);similarly,in the order of E?cadm/Ezrin(-),E?cadc/Ezrin(-),E?cadm/Ezrin(+),and E?cadc/Ezrin(+),the number of lymph node metastasis was increased significantly(P=0.007). Conclusion Ezrin may regulate the subcellular localization of E?cad in metastatic breast cancer ,which may affect the course of breast cancer and promote the metastasis of lymph nodes.

Objective To investigate the effect of extremely low frequency-electromagnetic field (ELF-EMF) on bone mesenchymal stem cells (BMSCs) differentiating into neuron like cells in vitro and research its mechanism.Methods BMSCs were collected from rats by means of whole bone marrow adherent.Flow cytometry was used to assay cell surface marker at passage 3.And then,BMSCs were assigned into four groups:ELF-EMF group,ELF-EMF+U0126 (inhibitor) group,U0126 group and control group;cells were induced by medium (2％ DMSO and 200 μmol/L BHA) for 5 h.In the process of neural induction,ELF-EMF group and ELF-EMF+U0126 group were received 10 Hz,500 GS ELF-EMF stimulation.Besides,ELF-EMF+U0126 group and U0126 group were pretreated with 50 μmol/L extracellular signal-regulated kinase (ERK)1/2 inhibitor U0126.The morphology of BMSCs was observed under inverted microscope.The expression of nestin was detected by immunofluorescent staining and Western blotting to identify and determine the differentiation.Western blotting was applied to detect the preotein level of phosphorylase-ERK1/2 after ELF-EMF exposure.Results BMSCs presented a single long spindle morphology,growing with close whirlpoor-like arrangement;CD90 expression rate was up to 97.9％,while that of CD45 only 4.7％.After induction,each group of cells showed similar shape with neuron-like cells gradually.As compared with the other three groups,ELF-EMF group had significantly higher expression levels of nestin and phosphorylatd ERK1/2 detected by immunofluorescent staining and Westem blotting,respectively (P＜0.05).Meanwhile,the expression levels of nestin among the ELF-EMF+U0126 group,U0126 group and control group were not statistically significant (P＞0.05).Conclusion ELF-EMF could promote neuronal differentiation of mesenchymal stem cells via activation of ERK1/2 signaling pathways.

Background:Functional dyspepsia(FD)is a commonly seen functional disease which has great impact on patient’s quality of life and mental health,and has got more and more concern by clinicians. Aims:To explore the role of mast cells (MC)and plasma motilin( MTL)in the pathogenesis of FD. Methods:Sixty FD patients including 32 postprandial distress syndrome(PDS)and 28 epigastric pain syndrome(EPS)patients from September 2013 to January 2014 at the Affiliated Hospital of Putian University were enrolled,and 28 healthy volunteers were served as controls(HS group). The number of gastric mucosal MC,plasma MTL level(fasting and 30 minutes after drinking warm water)and gastric emptying time(T1 / 2 )were compared. Results:Compared with HS group,number of gastric mucosal MC was significantly increased (P < 0. 001),plasma MTL level before and after drinking warm water in FD group were significantly decreased( P <0. 001),T1 / 2 was significantly prolonged(P < 0. 001). No significant difference in number of MC was found between PDS subgroup and EPS subgroup(P = 0. 094). Plasma MTL level before and after drinking warm water in PDS subgroup were significantly lower than those in EPS subgroup( P < 0. 001),and T1 / 2 was significantly prolonged( P < 0. 001). Conclusions:Number of gastric mucosal MC is significantly increased in FD patients compared with HS group,suggesting MC may play a role in the pathogenesis of FD. Plasma MTL level is decreased in FD patients,which may be one of the factors involved in the pathogenesis and occurrence of corresponding symptoms of FD.

Objective To investigate the diagnostic values of DCE-MRI quantitative evaluation of the activity of sacroiliac joints in ankylosing spondylitis.Methods Forty two patients (36 male and 6 female) with ankylosing spondylitis in our hospital were enrolled prospectively according to the standard of diagnostic criteria for ankylosing spondylitis revised in 1984.All of them were evaluated with the blood sedimentation,C-reactive protein and ankylosing spondylitis activity index (BASDAI).We divided the patients with ankylosing spondylitis into stationary phase group and active phase group.We analyzed each of the MRI images of the two patients groups using the extended Tofts model to determine the quantitative parameters of DCE-MRI,such as contrast transfer coefficient (Ktrans),reflux constant (Kep),extravascular extracellular volume fraction (Ve),plasma volume fraction (Vp).And then the differences of quantitative data between the two groups were compared.Spearman correlation analysis was performed between the parameters with statistically significant difference and BASDAI scores.Results Among the 42 patients,22 patients (18 male and 4 female) were in active phase group and the other 20 patients (18 male and 2 female) were in stationary phase group.Ktrans,Kep,Ve were (0.750± 1.130)/min,(1.008±0.732)/min,0.460± 1.735;(0.163±0.401)/min,(0.505 ± 0.902)/min,0.345 ±3.460 for active phase group and the stationary phase group,respectively.The results of the active phase group was significantly higher than the stationary phase group (Z=3.727,2.317,3.696,respectively;all P＜0.05).The Vp had no statistically significance for the active phase group (0.125 ± 0.310) and the stationary phase group (0.160 ± 0.329) (Z=1.209,P＞0.05).Strong correlations existed between Ktrans,Kep,Ve and BASDAI score,the correlation coefficients were 0.714,0.430,and 0.676 (P＜0.05).Conclusions Quantitative DCE-MRI parameters can evaluate the activity of the ankylosing spondylitis.Strong correlations exist between Ktrans,Kep,Ve and BASDAI score.

Objective To investigate the effect of di-(2-ethylhexyl) phthalate (DEHP) on the postnatal lung development in newborn rats.Methods A total of 60 newborn Sprague-Dawley rats (weighing 5.0-8.0 g) in five age groups were studied in the first experiment.The rats were divided based on the different postnatal ages:postnatal day (PND)I,PND4,PND7 and PND14.A total of 45 newborn Sprague-Dawley rats (weighing 5.0-8.0 g) were randomly divided into three groups according to the dosage of DEHP administered in the second experiment.The newborn rats were administered DEHP through intraperitoneal injection at 10 (low-dose subgroup),100 (medium-dose subgroup) or 750 (high-dose subgroup) mg/kg daily from PND1 to PND13.The rats were sacrificed on PND14.Pups were sacrificed with lethal dose injection of pentobarbital sodium.The lung was removed.The right middle lobes were used for analysis.The tissue was processed for histology and lung sections were stained with HE for light microscopic (LM) morphometric measurement.The analysis was performed by means of a digital image analysis system,including pulmonary interstitial area ratio (IAR) and total length density of all segments.One-way ANOVA,LSD and Dunnet T3 methods were used for statistical analysis.Results In the normal controls,IAR decreased significantly by (31.97±5.03) ％,(30.05±3.57)％,(25.33± 1.83)％ and(22.01 ±2.19)％,respectively,from PND1 to PND14 (P＜0.05 or P＜0.01).IAR in medium-and high-dose subgroups increased significantly by (24.11 ±2.78)％ and (26.53± 3.42)％,respectively on PND 14.The total length density of all segments in unit area lung volume increased significantly by 0.047 8±0.003 7,0.050 0±0.002 9,0.071 2±0.003 0 and 0.084 4±0.004 3,respectively from PND1 to PND14 (P＜0.01).In the DEHP treated animals,when compared with the control group,IAR was significantly higher on PND14 (P＜0.05 or P＜0.01),while the total length density of all segments in unit area lung volume was significantly decreased (P＜0.05 or P＜0.01).Length density in medium-and high-dose subgroups were higher than that of low-dose subgroup by 0.082 9±0.001 8,0.077 2±0.002 0 and 0.071 3±0.003 7,respectively on PND14 (P＜0.05 or P＜0.01).Conclusions Medium-and high-dose DEHP affect the postnatal lung development in rats in a dose-dependent mode.