This study aimed to investigate the therapeutic effects of Morinda officinalis iridoid glycosides (MOIG) on bone loss of rheumatoid arthritis (RA) rats, and the mechanism of osteoclast function and activity induced by lipopolysaccharide (LPS). RA rats were established by injecting bovin type II collagen. The Bio-ethic Committee of Zhejiang Chinese Medical University approved all experimental protocols associated with this study (IACUC-20180410-03). The collagen-induced arthritis (CIA) rats were administered drug by gavage for 8 weeks; the femoral trabecular micro-structure changes were observed in CIA rats by micro-CT; the LPS-induced osteoclasts model further observed the effect and mechanism of anti-inflammatory osteoporosis in vitro. The results indicated that MOIG could markedly increase bone mineral density (BMD) in CIA rats, improve trabecular micro-structure. In vitro studies demonstrated that MOIG could significantly inhibit osteoclastogensis and differentiation, suppress tartrate resistant acid phosphatase (TRAP) activity, F-actin ring formation, TNF receptor associated factor 6 (TRAF6) recruitment, and inhibitor of nuclear factor kappa-Bα (IκBα) degradation as well as p65 phosphorylation, thereby repressing nuclear factor kappa-B (NF-κB) signaling pathway activation. Subsequently, MOIG effectively inhibited osteoclast nuclear factor of activated T-cells c1 (NFATc1) and cellular oncogene Fos (c-Fos) expression, as well as bone resorption related protein activity including matrix metalloprotein 9 (MMP-9) and cathepsin K (CtsK). Meanwhile, MOIG also repressed the phosphorylation expression of Janus activating kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3), thereby inhibiting JAK2/STAT3 signaling pathway activation. Moreover, further studies found that MOIG could suppress glycogen synthase kinase-3β (GSK-3β) activity, and GSK-3β gene silencing could markedly inhibit oetsoclast F-actin ring formation as well as the phosphorylation expression of p65 and STAT3. Of note, compared with GSK-3β gene silencing group, there was no significant difference in the group treated with both MOIG with GSK-3β gene silencing simultaneously. Thus, the results suggested that MOIG may inhibit NF-κB signaling pathway and JAK2/STAT3 signaling pathway activation via regulating GSK-3β, thereby alleviating bone destruction in RA.

ObjectiveTo explore the role of structural MRI in the diagnosis of spinocerebellar ataxia type 3 （SCA3） and further evaluate its correlation with disease severity and disease duration. MethodsWe prospectively enrolled 81 genetically diagnosed SCA3 patients ［59 symptomatic （sym-SCA3） and 22 pre-symptomatic （pre-SCA3）］ and 35 age- and sex-matched healthy controls （HCs）. MRI structural images （3D T1 MPRAGE） and clinical data of all subjects were collected. Three observers with different radiological experience measured the width of the superior， middle and inferior cerebellar peduncle （SCP， MCP and ICP）， the anterior-posterior diameters of the pons and spinal cord at the levels of the foramen magnum and upper edge of the 3rd-5th cervical vertebra. One observer performed the measurements again 2 months later to assess for the intra- and inter-observer reliability， respectively. One-way ANOVA， rank-sum test， ROC curve and Random Forest were used to evaluate the diagnostic value of the above metrics for SCA3， and the correlation between the metrics and clinical variables was analyzed. ResultsNot depending on the radiological experience， the metrics based on morphological MRI showed high intra- and inter-observer reliability， among which bilateral superior and middle cerebellar peduncles performed best. The diameters of bilateral SCP， MCP， ICP， pons and spinal cord （except spinal cord at the level of the upper edge of the 5th cervical vertebra） decreased successively in HCs， pre-SCA3 and sym-SCA3 with a statistical difference （P<0.017）. ROC analysis revealed that the left MCP had the highest diagnostic value for pre-SCA3 （AUC=0.911）， with sensitivity， specificity and a cut-off value of 85.7%， 95.5% and 10.15 mm， respectively. In contrast， the right SCP had the highest diagnostic value for sym-SCA3 （AUC=0.999）， with sensitivity， specificity and a cut-off value of 100%， 98.3% and 2.62 mm， respectively. The Random Forest model based on the above metrics also had high diagnostic efficiency （AUC= 0.970， specificity=93.1%）， and the left MCP contributed the most. Correlation analysis showed that the above metrics had a significantly or moderately negative correlation with the Scale for the Assessment and Rating of Ataxia （SARA） and disease duration （P<0.05）. ConclusionNot depending on radiological experience， measurements of brain structure based on morphological MRI are reliable， which can help diagnose SCA3 and predict disease severity and duration. The left MCP and the right SCP perform best for predicting pre-SCA3 and sym-SCA3， respectively. Therefore， the structural MRI is recommended for assisting the clinical diagnosis of SCA3.

Alveolar bone is an important anatomic basis for implant-supported denture restoration, and its different degrees of defects determine the choices of bone augmentation surgeries. Therefore, the reconstruction of alveolar bone defects is an important technology in the clinical practice of implant restoration. However, the final reconstructive effect of bone quality, bone quantity and bone morphology is affected by many factors. Clinicians need to master the standardized diagnosis and treatment principles and methods to improve the treatment effect and achieve the goal of both aesthetic and functional reconstruction of both jaws. Based on the current clinical experience of domestic experts and the relevant academic guidelines of foreign counterparts, this expert consensus systematically and comprehensively summarized the augmentation strategies of alveolar bone defects from two aspects: the classification of alveolar bone defects and the appropriate selection of bone augmentation surgeries. The following consensus are reached: alveolar bone defects can be divided into five types (Ⅰ-0, Ⅰ-Ⅰ, Ⅱ-0, Ⅱ-Ⅰ and Ⅱ-Ⅱ) according to the relationship between alveolar bone defects and the expected position of dental implants. A typeⅠ-0 bone defect is a bone defect on one side of the alveolar bone that does not exceed 50% of the expected implant length, and there is no obvious defect on the other side; guided bone regeneration with simultaneous implant implantation is preferred. Type Ⅰ-Ⅰ bone defects refer to bone defects on both sides of alveolar bone those do not exceed 50% of the expected implant length; the first choice is autologous bone block onlay grafting for bone increments with staged implant placement or transcrestal sinus floor elevation with simultaneous implant implantation. Type Ⅱ-0 bone defects show that the bone defect on one side of alveolar bone exceeds 50% of the expected implant length, and there’s no obvious defect on the other side; autologous bone block onlay grafting (thickness ≤ 4 mm) or alveolar ridge splitting (thickness > 4 mm) is preferred for bone augmentation with staged implant placement. Type Ⅱ-Ⅰ bone defects indicate that the bone plate defect on one side exceeds 50% of the expected implant length and the bone defect on the other side does not exceed 50% of the expected implant length; autologous bone block onlay grafting or tenting techniques is preferred for bone increments with staged implant implantation. Type Ⅱ-Ⅱ bone defects are bone plates on both sides of alveolar bone those exceed 50% of the expected implant length; guided bone regeneration with rigid mesh or maxillary sinus floor elevation or cortical autologous bone tenting is preferred for bone increments with staged implant implantation. This consensus will provide clinical physicians with appropriate augmentation strategies for alveolar bone defects.

Aim To explore the effect of endoplasmic reticulum (ER) stress-related exosomes derived from hepatocellular carcinoma (HCC) cells on the M2 polarization of macrophages via Toll-like receptor 4 (TLR4) and the underlying mechanism. Methods Western blot experiments were conducted to evaluate the GRP78 levels in HCC tissues and HCC cells with or without pre-treatment of tunicamycin and the identification of the isolated exosomes. Immunohistochemical staining was also employed to detect the expression of GRP78 in HCC tissues. Immunofluorescence was used to confirm the effective absorbance of exosomes by macrophages. Flow cytometry ( FCM ) , Western blot and CBA kit were used to detect M2 polarization-asso ciated markers and the expression of TLR4 on macrophages. Affymetrix gene chip and bioinformatics analy-sis were used to screen the differential expression of mRNA in macrophages stimulated by ER stressed-relat- ed exosomes and TLR4 expression. Results In clinical samples, GRP78 expression in liver cancer tissues was higher than that in the adjacent tissues. In cell experiments, tunicamycin (2.5 jxmol • L"1 ) treatment for 24 hours in HCC cells could significantly increase the expression of ER stress marker protein GRP78 compared with control group. Western blot showed that HCC cell-derived exosomes positively expressed exosom- al marker proteins,such as CD63,TSG101 and HSP70, and negatively expressed Calnexin. Interestingly, HSP70-enriched exosomes released by ER-stressed HCC cells were observed. Laser confocal microscopy found that exosomes could be effectively taken up by RAW264.7 macrophages. Flow cytometry and Western blot further indicated that ER stress-related exosomes could significantly up-regulate the expression of CD206, transformed growth factor-beta (TGF-p) and arginase-1 (Arg-1) in macrophages. Meanwhile, these exosomes could promote the secretion of IL-6 and IL- 10,and up-regulate the expression of TLR4 in macrophages. Conclusions ER stressed-HCC cells could promote macrophage M2 polarization by releasing HSP70-enriched exosomes and activating TLR4 signaling.

Background:Transcranial alternating current stimulation (tACS) offers a new approach for adult patients with major depressive disorder (MDD).The study is to evaluate the efficacy and safety of tACS treating MDD.Methods:This is an 8-week,double-blind,randomized,placebo-controlled study.Ninety-two drug-naive patients with MDD aged 18 to 65 years will receive 20 daily 40-min,77.5-Hz,15-mA sessions of active or sham tACS targeting the forehead and both mastoid areas on weekdays for 4 consecutive weeks (week 4),following a 4-week observation period (week 8).The primary outcome is the remission rate defined as the 17-item Hamilton depression rating scale (HDRS-17) score ≤7 at week 8.Secondary outcomes are the rates of response at weeks 4 and 8 and rate of remission at week 4 based on HDRS-17,the proportion of participants having improvement in the clinical global impression-improvement,the change in HDRS-17 score (range,0-52,with higher scores indicating more depression) over the study,and variations of brain imaging and neurocognition from baseline to week 4.Safety will be assessed by vital signs at weeks 4 and 8,and adverse events will be collected during the entire study.Discussion:The tACS applied in this trial may have treatment effects on MDD with minimal side effects.

OBJECTIVETo explore the anti-myeloma effect of suberoylanilide hydroxamic acid (SAHA) and on mouse myeloma cell line SP2/0 in vitro and in vivo and its mechanism.

METHODSThe inhibitory effect of SAHA on SP2/0 cells was measured by CCK-8 assay,and the apoptosis and cell cycle were analyzed by flow cytometry FACS. The protein expression of Caspase-3 and p53 of SP2/0 cells treated with SAHA were examined by Western blot. Annexin V/7-AAD double staining was performed to detect the apoptosis of SP2/0 induced by SAHA in vitro. SP2/0 cells (1×10) resuspended in 200 µl PBS were inoculated subcutaneously and intravenously into BALB/c mice, so as to establish aggressive or non-aggressive myeloma-bearing mouse models respectively. On day 3 after modeling, mice received SAHA or vehicle control treatment by intraperitoneal injection. The dose of SAHA was 60 mg/kg·d, 5 times a week for 3 weeks.

RESULTSIn SAHA-treated SP2/0 cells, the proliferation inhibition rate and apoptotic cells increased in a dose dependent manner. Also, SAHA significantly increased the ratio of cells in G phase and decreased in S phase. Molecular mechanisms of apoptosis and cell cycle arrest of SP2/0 induced by SAHA partly correlated with up-regulating the expression level of Caspase-3 and p53. In the non-aggressive myeloma-bearing mice, SP2/0 cells disappeared in peripheral blood after SAHA treatment. In the aggressive myeloma-bearing mice, inhibition of tumor growth and prolongation of the cell survival were observed after SAHA treatment.

CONCLUSIONSAHA inhibited SP2/0 cell proliferation, this effect associates with inducing apoptosis and cell cycle arrest, the mechanism of SAHA ralates partly with activating Caspase-3 and p53 pathway.

Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Histone Deacetylase Inhibitors ; Hydroxamic Acids ; Mice ; Mice, Inbred BALB C ; Multiple Myeloma

[Objectives]To observe the clinical effects of the immediate implant placement of Zimmer dental implant sys-tem in the mandibular posterior region.[Methods]67 cases with a total of 89 mandible posterior teeth to deal with immediate implantion which selected to be treated with Zimmer dental implant system.76 teeth between implant and tooth socket bone wall were simultaneously filled with Bio-oss Collagen.The upper structure was repaired with PFM porcelain crown after the postoperative phase I of 3~6 months.All the patients were followed up for 6~24 months.[Results]During the clinical follow up,the implant survival rate was 100%. In 67 patients,89 implants was successfully loaded,with stable implants,good condition in synosteosis and without adverse subjective symptoms. All of the 67 patients had achieved good synosteosis and success loads clinically and radiologically.[Conclusion]A good osseointegration is obtained in the mandibular posterior re-gion with Zimmer dental implant system,Correctly dealing with Bio-oss Collagen between implant and tooth socket bone wall.At the same time,it can shorten the time of therapy,simplify the procedure,so that the clinical results are more satis-factory.

Objective To analyze the complications of coronary rotational atherectomy and to evaluate the safety of the procedure. Methods We evaluated the procedural and angiographic outcomes of 248 consecutive procedures to rotational atherectomy between January 2000 and October 2016. Results 27 cases(10.9％)were found to have rotational atherectomy related complications. Among these 27 cases,coronary spasm occurred in 2 cases(0.8％),no reflow in 8 cases(3.2％), coronary dissection in 5 cases(2.0％),burr entrapment in 6 cases(2.4％),wire breakage in 3 cases(1.2％), and coronary perforation in 3 cases(1.2％). There were no death,acute myocardial infarction and emergent coronary bypass graft.14 case(5.6％)had PCI-related myocardial infarction during hospital stay.In-hospital major adverse cardiaccerebral event(MACCE)rate was 5.6％. Conclusions Coronary rotational atherectomy can be performed with high success rates and procedure-related complications are rare.

The study aims to investigate the associations of SUMO4 polymorphisms with tacrolimus concentrations in Chinese renal transplant recipients. Blood samples and clinical data were collected from 132 renal transplant recipients with tacrolimus treatment. CYP3A5*3 genotypes were detected by PCR-RFLP, and SUMO4 (rs237024, rs237025) genotypes were detected by Sequenom® MassARRAY system. SUMO4 rs237024 and rs237025 genotypes were in complete linkage disequilibrium (D' = 1). The dose-adjusted concentration of tacrolimus in SUMO4 rs237024A-rs237025A (GA-GA +AA-AA) carriers was considerably higher than that in GG-GG carriers (P < 0.05). After stratification by CYP3A5*3 genotypes, SUMO4 rs237024A-rs237025A carriers (GA-GA+AA-AA) had a higher dose-adjusted tacrolimus concentration than that in GG carriers in CYP3A5 expresser (P < 0.05). The results illustrated that SUMO4 rs237024 and rs237025 polymorphisms were associated with tacrolimus concentrations, and the test of these genotypes may be useful for individualized medicine of tacrolimus.
Asian Continental Ancestry Group ; genetics ; Cytochrome P-450 CYP3A ; genetics ; Genotype ; Humans ; Immunosuppressive Agents ; blood ; therapeutic use ; Kidney ; Kidney Transplantation ; Linkage Disequilibrium ; Polymorphism, Single Nucleotide ; Small Ubiquitin-Related Modifier Proteins ; genetics ; Tacrolimus ; blood ; therapeutic use