Objective Establishment of two experimental models for osteoclast differentiation from monocyte in vitro,and to study the potential of osteoclast differentiation induced by cytokines.Methods Direct model of osteoclast differentiation: CD14+ monocyte fraction of peripheral blood mononuclear cell(PBMC) stimulated by(25 ?g/L) M-CSF+(10~(-8)mol/L) LTB4 for two weeks.Indirect model of osteoclast differentiation: Utilize the coculture model of RAFLs and monocyte that were stimulated in the presence of 25 g/L M-CSF+(10~(-8)mol/L) LTB4 for three weeks.In TRAP staining the multinucleated TRAP staining positive osteoclast-like cells were counted as marker of as differentiation effect of each group.Results Osteoclast-like cells can be induced by both direct and indirect models.Conclusion Two experimental models for osteoclast differentiation can be separately used to study the effect of various cytokines for direct and indirect OC differentiation.

Aim To investigate whether nicorandil (Nic)protects H9c2 cardiac cells against high glucose (HG)-induced injury and inflammation by inhibiting nuclear factor-κB (NF-κB )/cyclooxygenase-2 (COX-2 )pathway.Methods Cell viability was measured by cell counter kit-8 (CCK-8)assay.The expression lev-els of NF-κB,COX-2 and cleaved caspase-3 were de-termined by Western blot.The activity of lactate dehy-drogenase (LDH)in the culture medium was measured with commercial kits.The intracellular level of reactive oxygen species (ROS)was detected by 2′,7′-dichlor-fluorescein-diacetate (DCFH-DA)staining followed by photofluorography.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography.Mitochondrial membrane poten-tial (MMP)was examined by rhodamine 123 staining followed by photofluorography.The secretion levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were detected by ELISA.Results After H9 c2 cardiac cells were treated with 35 mmol · L-1 glucose (high glucose,HG)for 24 h,the cell viability was significantly decreased .Pre-treatment of the cells with 20~100 μmol·L-1 Nic for 60 min or 50 μmol· L-1 Nic for 30~120 min before exposure to HG signif-icantly attenuated the decrease in viability induced by HG.On the other hand,HG increased the expression levels of phosphorated (p)-NF-κB p65 and cyclooxy-genase-2 (COX-2 )in H9c2 cardiac cells.Pre-treat-ment of the cells with 50 μmol·L-1 Nic for 60 min at-tenuated the up-regulation of p-NF-κB p65 and COX-2 expression levels induced by HG.Furthermore,HG induced considerable injuries and inflammatory re-sponse,leading to increases in LDH activity,ROS generation,MMP loss,the number of apoptotic cells, the expression of cleaved caspase-3 as well as the se-cretion levels of IL-1βand TNF-α.Pre-treatment of the cells with 50 μmol·L-1 Nic for 60 min before HG exposure,or co-treatment of the cells with 100 μmol· L-1 PDTC (an inhibitor of NF-κB)or 10 μmol·L-1 NS-398 (an inhibitor of COX-2)and HG for 24 h ob-viously reduced the above injuries and inflammatory re-sponse induced by HG. Conclusion Nic protects H9 c2 cardiac cells against HG-induced injury and in-flammation by inhibiting NF-κB/COX-2 pathway.

AIM:To investigate whether angiotensin-(1-7) [Ang-(1-7)] protects H9c2 cardiac cells against high glucose (HG)-induced injury and inflammation by inhibiting the interaction between Toll-like receptor 4 (TLR4) acti-vation and necroptosis .METHODS:The expression levels of receptor-interacting protein 3 ( RIP3;an indicator of necrop-tosis) and TLR4 were determined by Western blot .Cell viability was measured by CCK-8 assay.The activity of lactate de-hydrogenase ( LDH) in the culture medium was measured with a commercial kit .The releases of interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α) were measured by ELISA .The intracellular level of reactive oxygen species ( ROS) was analyzed by 2 ’ , 7 ’-dichlorfluorescein-diacetate ( DCFH-DA ) stating followed by photofluorography .Mitochondrial membrane potential ( MMP) was examined by rhodamine 123 staining followed by photofluorography .RESULTS:After the H9c2 cardiac cells were treated with HG (35 mmol/L glucose) for 24 h, the expression of RIP3 was obviously increased . Co-treatment of the cells with 30μmol/L TAK-242 (an inhibitor of TLR4) attenuated the up-regulation of RIP3 induced by HG.Furthermore, the expression of TLR4 was significantly increased after the cells were exposed to HG for 24 h, and co-treatment of the cells with 100μmol/L necrostatin-1 ( Nec-1;a specific inhibitor of necroptosis ) and HG for 24 h attenua-ted the up-regulation of TLR4 expression induced by HG .Moreover, 1μmol/L Ang-(1-7) simultaneously blocked the up-regulation of the RIP3 and TLR4 induced by HG.On the other hand, co-treatment of the cells with 1μmol/L Ang-(1-7), 30 μmol/L TAK-242 or 100 μmol/L Nec-1 and HG for 24 h attenuated HG-induced injuries and inflammatory response , leading to the increase in the cell viability , and the decreases in the activity of LDH , ROS generation , MMP loss as well as the releases of IL-1βand TNF-α.CONCLUSION:Ang-(1-7) protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the interaction between TLR 4 activation and necroptosis .

AIM:Tostudywhe ther theangiotens in-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardio-myocytes against high glucose (HG)-induced injury by inhibiting nuclear factor-κB (NF-κB) pathway.METHODS:The cell viability was measured by CCK-8 assay.The intracellular levels of reactive oxygen species ( ROS) were detected by DCFH-DA staining .The number of apoptotic cells was tested by Hoechst 33258 nuclear staining .Mitochondrial membrane potential ( MMP) was examined by JC-1 staining.The levels of NF-κB p65 subunit and cleaved caspase-3 protein were de-termined by Western blotting.RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose (HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated ( p) NF-κB p65, peaking at 60 min.Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30μmol/L and HG for 24 h inhibited HG-induced cy-totoxicity, evidenced by an increase in cell viability .On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage , indicated by decreases in the number of apoptotic cells , cleaved caspase-3 level, ROS generation and MMP loss .However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor (Mas receptor).Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG.CONCLUSION:Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway .

Objective To investigate the damage sequence of posterior ligamentous complex (PLC) and its clinical significance in thoracolumbar fracture.Methods Data of 132 patients with spinal fracture evaluated with X-rays,CT and short-tau inversion-recovery (STIR) sequences in MRI were collected prospectively.Fracture morphology was classified using the AO classification.PLC components including interspinous ligament (ISL),supraspinous ligament (SSL),ligamentum flavum (LF) and facet capsules (FC) were assessed and classified as intact,edema,or tear.ISL edema was further subdivided depending on the extension (＞ 50％ or ≤50％).Correlation between MRI signal and AO progressive scale of morphological damage was analyzed.Results AO type A1/A2 fracture associated with only FC distraction.AO type A3 fracture showed additional ISL tear,usually less than 50％,with neither LF nor SSL tear.AO type B1 fracture showed FC distraction,ISL edema or disruption,and low rate of SSL/LF tear,but B2 fracture increased the rate of SSL/LF tear.AO type C fracture showed facet fracture or dislocation and ISL,SSL as well as LF tear.High correlation was found between AO progressive scale and MRI signal (P ＜ 0.01).Conclusions MRI study can well display the PLC damage and damage sequence.MRI correlates with AO progressive scale of morphological damage,which shows a progressive orderly rupture sequence among different PLC components as traumatic forces increase.

Objective:To examine the clinical value of combining indocyanine green (ICG) fluorescence navigation with blue dye in sen-tinel lymph node biopsy (SLNB) for patients with breast cancer. Methods:A total of 89 patients with early-stage breast cancer who met the inclusion criteria were admitted at Shantou Central Hospital, Guangdong from May 2013 to April 2014. In phase one, ICG and blue dye were applied in all 53 patients, and then SLNB and axillary lymph node dissection (ALND) were performed based on fluores-cence signal or visual sense of the lymph nodes. In phase two, 36 patients with early-stage breast cancer were included. ALND was omitted when sentinel lymph nodes were frozen showing negative result. Rates of detection, accuracy, and false-negative were calcu-lated. Results:A total of 89 patients were monitored, of which the total rate of SLNB detection was 96.6%(86/89). In the validation pe-riod, the rates of detection, accuracy, and false-negative were 94.3%(50/53) 98.0%(49/50), and 2.6%(1/38), respectively. In the alter-ative period, the rates of detection reached 100%. Of the 196 sentinel lymph nodes, 179 showed fluorescence signal, 142 exhibited blue dying, 54 only demonstrated fluorescence signals, and 45 demonstrated metastasis with five signaling fluorescence. About 24.7%of patients were diagnosed with SLN metastasis (22/89), where SLNB in two patients showed fluorescence signal but without blue dye. No ipsilateral lymph node relapsed were observed during a median follow up of 25 months. Conclusion:Combination of ICG fluores-cence navigation with blue dye in SLNB is safe for patients with breast cancer.

Objective To investigate the peripheral blood neutrophil to lymphocyte ratio(NLR) for the early diagnosis of acute coronary syndrome(ACS) .Methods A total of 247 patients with suspected ACS and chest pain ,including 51 cases with acute ST segment elevation myocardial infarction(STEMI) ,42 cases with acute non‐ST segment elevation myocardial infarction(NSTEMI) , 87 cases with unstable angina pectoris(UA) and 67 cases with non‐cardiogenic chest pain(NCCP)were enrolled and detected for white blood cells count and classification .The sensitivity ,specificity ,positive predictive value ,negative predictive value ,receiver op‐erating characteristic(ROC) curve of NLR were analyzed .Results Among all patients ,the most common was UA ,followed by NC‐CP ,STEMI and NSTEMI .Level of neutrophil proportion and white blood cell count were lowest in NCCP group ,followed by UA , NSTEMI and STEMI group ,but lymphocyte proportion was with the opposite change tendency .Diagnostic sensitivity ,specificity , accuracy ,positive predictive value and negative predictive value of NLR for ACS were higher than white blood cell count .Conclusion NLR was with various advantages for the early diagnosis ,prognosis evaluation and state of ACS .

Objective To investigate the effect of coenzyme complex on the cardiac and renal functions of patients with type 2 cardiorenal syndrome. Methods Sixty-two patients with type 2 cardiorenal syndrome were enrolled in from June 2013 to December 2014 in Zhujiang Hospital , Southern Medical University. These patients divided were into routine group (n = 31) and coenzyme complex (n = 31). The therapy scheme of coenzyme complex group was on the basis of routine group with coenzyme complex intravenous drip , 400 U/day for 2 weeks. The cardiac function was determined by New York Heart Function Assessment, and the cardiac ultrasound, the levels of BNP. The renal function was determined by serum creatinine and urine volume. Results Compared with routine group, the rates of NYHA Ⅲ and Ⅳ were reduced, the LVEF levels were increased and the levels of BNP were increased (P < 0.05). The serum creatinine was decreased and urine volume were increased in the coenzyme complex group (P<0.05). Conclusion Coenzyme complex could improve the cardiac and renal functions of the patients with type 2 cardiorenal syndrome.

Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in the adult urban population of Hezhou Guangxi. Methods One thousand and two hundred urban residents (older than 18 years) from Hezhou Guangxi were randomly selected using a random sampling. All the residents were interviewed. Their morning spot urine were tested to determine albumin to ereatinine ratio (abnormal:≥30 mg/g), and renal function [abnomal: eMDRD <60 ml·min-1·(1.73 m2)-1] was assessed. Morning spot urine dipstick of hematuria (abnormal:≥1 +) was confirmed by microscopy (abnormal: 3 red blood cells/HP). The associations among demographic characteristics, health eharacteristies and indicators of kidney damage were examined. Results Eligible data of 1069 subjects were enrolled in the study. The prevalence of albuminuria was 7.5%, hematuria 4.8%, and reduced eGFR 3.6%. The prevalence of kidney disease was 14.4% and the recognition was 1.4%. Age (OR 1.022, 95%CI 1.008-1.035), gender (OR 2.249, 95%CI 1.502-3.367), diabetes mellitus (OR 7.422, 95%CI 3.985-13.825) and hypertension (OR 4.397, 95% CI 2.601-7.432) were independently associated with CKD. Conclusions The prevalence of chronic kidney disease is 14.4% and the recognition is 1.4% in adult urban population of Hezhou Guangxi. Independent risk factors associated with chronic kidney disease are age, gender, diabetes mellitus and hypertension which is similar to those in developed countries and domestic big cities.

Objective :To explore influence of ticagrelor on levels of serum high sensitive C reactive protein (hsCRP) and plasma homocysteine (Hcy) in patients with acute coronary syndrome (ACS).Methods :A total of 135 ACS pa‐ tients hospitalized in our department from Jan 2016 to Feb 2017 were selected .Based on routine treatment ,Patients were randomly and equally divided into routine group ,clopidogrel group and ticagrelor group (based on routine treatment respectively received clopidogrel or ticagrelor ) for four weeks .Levels of serum hsCRP and plasma Hcy were measured and compared among all groups before and after treatment .Results :Compared with before treat‐ment ,after four‐week treatment , there were significant reductions in levels of serum hsCRP and plasma Hcy in three groups (P＜0. 05 or ＜0.01).Compared with routine group and clopidogrel group after four‐week treatment , there were significant reductions in levels of serum hsCRP [ (12.95 ± 1.99) mg／L , (8. 56 ± 1. 24) mg／L vs.(4. 47 ± 1. 92) mg／L] and plasma Hcy [ (13.48 ± 2.12) μmol／L , (9.55 ± 0. 94) μmol／L vs.(6. 61 ± 1. 15) μmol／L] in ticagrelor group ( P＜0.05 or ＜0.01).Conclusion :Ticagrelor can significantly reduce levels of serum hsCRP and plasma Hcy while effective antiplatelet therapy ,then significantly inhibit inflammatory response ,improve vascular endothelial function ,contribute to stabilizing atherosclerotic plaques ,improve prognosis in ACS patients .