AIM:To explore the effect of leptin on the expression of degeneration-related genes in rat nucleus pulposus ( NP) cells and to detect the possible mechanism .METHODS:The normal NP cells isolated from SD rats were analyzed by immunochemistry and immunofluorescence for the collagen II and cytokeratin 19 expression.The NP cells were treated with leptin and/or interleukin-1β( IL-β).The mRNA expression of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, ADAMTS-5, aggrecan and COL2A1 in the cells was detected by real-time PCR.Alcian blue staining and im-munochemistry were used to examine the expression of proteoglycan and collagen II .Activation of involved pathways was studied by Western blot .The inhibitors of the pathways were used to reveal the effect of these pathways on NP cells .RE-SULTS:The results of real-time PCR revealed that leptin alone up-regulated the mRNA expression of MMP-1, MMP-13, ADAMTS-4, ADAMTS-5 and COL2A1.The synergy of leptin and IL-βwas found in the increased expression of MMP-1, MMP-3 and ADAMTS-5.The NP cells treated with leptin showed less expression of collagen II .Both PI3K/Akt and JAK2/SATA3 pathways were activated by leptin , whereas only inhibitor of JAK 2/SATA3 pathway reversed the expression of MMP-1 and MMP-13.CONCLUSION:Leptin may promote catabolism in rat NP cells via JAK2/SATA3 pathways, which may be the mechanism mediating the association between obesity and intervertebral disc degeneration .

AIM:Toexploretheeffectofglucocorticoidonautophagyandsenescenceinthechondrocytes. METHODS:The collagen II in the normal chondrocytes isolated from the SD rats was checked.After stimulation with glu-cocorticoid, LysoTracker Red staining, MDC staining and Western blot were used to detect the level of autophagy in the chondrocytes.The mTOR pathway related molecules were investigated by Western blot.Cell senescence was analyzed by SA-β-gal staining.RESULTS:A dose-dependent increase in the number of autophagic vacuoles was observed in the dexa-methasone-treated chondrocytes, which was demonstrated by the LysoTracker Red and MDC staining.The expression of LC3-II and beclin-1 was increased by dexamethasone, especially in the cells treated with dexamethasone for 4 d.However, P62 expression was decreased.SA-β-gal staining showed that the percentage of cell senescence was increased by dexam-ethasone.Surprisingly, the cell senescence induced by dexamethasone was exacerbated by the autophagic inhibitor 3-MA. CONCLUSION:Autophagy induced by dexamethasone protects chondrocyte from senescence.The mTOR pathway may be involved in the autophagy activation.

Objective To investigate the effectiveness of entropy in measuring the level of sedation during target controlled infusion (TCI) of propofol in patients of different ages. Methods Twenty-nine ASA Ⅰ-Ⅱ patients undergoing elective surgery under general anesthesia were divided into two age groups: Ⅰ adult group (20-64 yr) (n = 16) and Ⅱ the aged group (65-85 yr) ( n = 13). The patients were unpremedicated. The level of sedation was assessed using OAA/S scale. Propofol was given by TCI. The effect-site concentration (Ce) of propofol was set first at 1 ?g?ml-1 and then increased step by step by 0.5 ?g?ml-1 every 60 seconds until Ce reached 6 ?g?ml-1. Response entropy ( RE), state entropy ( SE), BP, HR, SpO2 were monitored and recorded at each Ce, before intubation, and immediately and 1, 2, 3, 4, 5, 10 min after intubation. The predictive performance of entropy was evaluated by prediction probability (Pk) .Results The two groups were comparable with respect to sex (M/F ratio), body weight and body weight index (kg?m-2). The RE and SE values decreased as Ce increased. The difference between RE and SE was also reduced. In adult group when Ce reached 2.0 ?g?ml-1, the RE and SE values were lower than the baseline values ( P

Dexmedetomidine( DEX) is a pure potent, highly se-lective and highly specific agonist ofα2-adrenergic receptors with sedative, analgesic and sympatholytic properties. The sedative effect mimics natural sleep of“arousable” and“cooperative” se-dation without respiratory depression. Due to the above properties and advantages, DEX has received adequate attention in clinical practice and its spectrum of application is also expanding. In re-cent years, it is proved that DEX is neuroprotective not only in animal researches but also in clinical studies. The neuroprotec-tion of DEX and its related mechanism will be briefly reviewed in this paper.

Objective To investigate the effect of the combination therapy of tirofiban and reteplase on endothelial function,coagulation function and plaque inflammation in elderly patients with ST-elevation acute myocardial infarction (STEMI).Methods 100 patients with STEMI were treated with percutaneous transluminal coronary intervention (PCI) from January 2014 to June 2016 in our hospital.39 cases in the control group used conventional oral aspirin,clopidogrel and statins and other treatment.61 cases in the observation group received tirofiban and reteplase on the basis of the control group.The expression of endothelial microparticles (EMP) was detected by flow cytometry (FCM),and the ICAM-1,high sensitivity C-reactive protein (hs-CRP),tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6),endothelin-1(ET-1) were measured by enzyme-linked immunosorbent assay (ELISA).The thrombin time (TT),activated partial thrombin time (APTT),prothrombin time (PT) and other indicators were measured by PUN-2048A coagulation instrument,then statistical analysis was performed.Results The postoperative levels of EMP,ICAM-1 and ET-1 of control group were (693.46±90.72),(768.58±20.46)μg/L and (31.27±8.18)ng/L,which were significantly higher than those in the observation group [(652.36±67.39),(752.37±25.0)μg/L,(28.22±5.05)ng/L],the differences were statistically significant (t=2.41,2.67,2.68,all P<0.05).After operation,the hs-CRP,TNF-α,IL-6 levels in the control group were (4.16±2.35)mg/L,(4.32±2.02)ng/L,(10.59±3.16)ng/mL,which were significantly higher than those in the observation group [(2.22±1.47)mg/L,(2.74±1.52)ng/L,(6.33±2.24)ng/mL],the differences were statistically significant(t=2.65,2.67,3.42,all P<0.05).The postoperative TT,PT,APTT in the observation group were (26.31±3.18)s,(14.34±1.67)s,(27.20±4.12)s,which were significantly longer than those in the control group [(24.03±2.84)s,(12.56±1.43)s,(24.55±3.62)s],the differences were statistically significant(t=2.15,2.31,2.65,all P<0.05).Conclusion Tirofiban combined with reteplase can improve endothelial function,inhibit inflammatory reaction and regulate coagulation function.

Objective To investigate the effects of vitamin D deficiency on pregnancy and fetal development in mice. Methods All female ICR mice were randomly divided into two groups. In control group, mice were fed with standard feeds (vitamin D3 >800 IU/kg). In vitamin D deficiency group (VDD), female mice were fed with feeds with vitamin D depletion (vitamin D3 <25 IU/kg). For mating purposes, four females were housed overnight with two males starting at 9 : 00 PM. Females were checked 7 : 00 AM the next morning, and the presence of a vaginal plug was designated as gestational day (GD) 0. All pregnancy mice were sacrificed on GD18. For each litter, the number of live fetuses, dead fetuses and resorption sites were counted. Live fetuses in each litter were weighed. Crown-rump lengths were examined. And skeletal development of all live fetuses in each litter was evaluated. Re-sults The pregnancy rate was lower in VDD group than that in control group(P<0.05). The number of live fetu-ses was significantly decreased(P<0.05). By contrast, the numbers of dead fetuses and resorption sites were sig-nificantly increased(P<0.05). In addition, the average weights of fetuses and placenta were reduced in vitamin D deficiency group. Moreover, vitamin D deficiency resulted in skeletal development retardation in fetuses. Conclu-sion Vitamin D deficiency impairs pregnancy and developmental outcomes in mice.

Objective To investigate and analyze the postoperative progression of ossification of the patients with ossification of the posterior longitudinal ligament (OPLL) on cervical spine. Methods From Jaunary 2001 to December 2007, 95 postoperative patients with cervical OPLL were followed and analyzed retrospectively. There are 72 males, 23 females, with the average age of 56.3 years (range, 40-73years). The follow-up time was from 1 to 6 years, average 3.1 years. Among them 36 patients were performed with anterior cervical corpectomy, fusion with titanium mesh and fixed with cervical plates, others treated with posterior cervical laminectomy and fixation. 2 of the 95 cases were performed anterior and posterior combined operation. Clinical data, X-rays, CT and MR images and progression of ossification, were measured and analyzed in details .The relationships between the progression of ossification and relative factors, as gender, age, C3 ossified involved, T-OPLL, OPLL-type, time of follow-up, surgical approach, Japanese Orthopaedic Association (JOA) scores and improvement rate of JOA scores, were analyzed. Results Progression of ossification in 39 cases among the 95 followed postoperative OPLL patients, 28 men and 11 women, average age 55.9 years, range 41-71 years. The age of progressed patients included 12 cases of ≤49 years, 12of 50-59 years, 12 of 60-69 years and 3 of ≥70 years. 35 patients were operated by posterior approach and only 4 treated with anterior operation. According to the standard of the progression of ossification that 2 mm in the length or/and thickness, there are 4 cases progressed only in length, 2 only in thickness, other 33 patients both the length and thickness. Progression of length is from 2mm to 20mm (average 7.74±4.71). But thickness is progressed from 2 mm to 6 mm (average 2.67±1.51). From 1 to 3 years follow-up time it appeard as a downtrend about the progression of ossification. But it may appear an uptrend from the 4th year. JOA score and improvement rate of the JOA score were almost improved to the high-point in three years. And according to the statistic data there are obvious relationship between progression of ossification with age, surgical approach and C3 ossified involved. Conclusion There is a high rate of postoperative ossification progression in cervical OPLL patients. Cervical OPLL patients with C3 ossification involved, performed with posterior laminectomy and those young at surgery may have higher rate of progression of the ossification. The JOA score and improvement rate of the JOA score were little influenced by the progression of the OPLL during the short and intermediate-term follow-up.

Objective　To construct an anti-LPS single chain phage antibody library in mice for further biomedical works. Methods　Total RNA was extracted from splenic cells for reverse transcription after BALB/C mice had been immunized with pure LPS for 4 weeks. The designed primers were used to amplify the variable region genes of both heavy and light chain (VH,VL) with polymerase chain reaction. The VH and VL were then conjugated to form a single chain of variable fragment (ScFv) by a linker. The ScFv was cut by NotⅠ and SfiⅠ and then ligated into a pCANTAB5E phagemid vector which was digested with the same restriction enzymes. The ligated vector was then tranfected into the competent E.coli TG1 cells. Four TG1 clones were randomly selected to detect the exotic DNA. Results　The titer of anti-LPS in murine sera was 1∶12 800. The concentration of total RNA was 12.3813 μg/ml. The length of the fragments were 340 bp for VH, 320 bp for VL and 800 bp for ScFv respectively. 1.9×107 clones were grown after transfection and one from the four randomly selected clones was identified to contain the exotic DNA. Conclusion　A 4.75×106 murine anti-LPS single chain phage antibody library was successfully constructed.

BACKGROUND:Ricin possesses strong cytotoxicity but lacks of specificity.Severe adverse reactions can be caused if ricin is systemically administered.Interstitial chemotherapy with ricin coated by a sustained-release carrier can enhance local therapeutic effects of tumor,avoiding systemic-toxic occurrence.OBJECTIVE:To prepare injectable and biodegradable thermosensitive poly(D,L-lactide-co-glycolide)(PLGA)-polyethylene glycol(PEG)-PLGA copolymer-based hydrogels and to observe the in vitro drug delivery of the PLGA-PEO-PLGA copolymer hydrogel as a sustained-release carrier of ricin.DESIGN,TIME AND SETTING:A controlled observation was performed at the Center for Medical Bioengineering.Fujian Medical University between August and December 2007.MATERlALS:Ricin with a purity of 95%was purified by Fuiian Institute of Ultrasonic Medicine.DL-lactide and L-glycolide were provided by Glycolide Lactide Co.,Ltd..China.METHODS:PLGA-PEG-PLGA-copolymer was synthesized by ring-opening polymerization.MAIN OUTCOME MEASURES:Phase transition temperature was determined by test tube turnover.relativemolecular mass by high-performance gel permeation chromatography,the proportion of block copolymer by nuclear magnetic resonance spectrometry.the in vitro drug delivery of ricin coated by PLGA-PEG-PLGA copolymcr hydrogelby enzyme linked immunosorbent assay.the structural integrity of released ricin by high-performance liquid chromatography,the cytotoxicity of released ricin by methyl thiazolyl tetrazolium assay.RESULTS:The synthesized copolymer PLGA-PEG-PLGA exhibited reverse.thermosensitivity.The phase transition temperature of the copolymer was 26℃at 230 g/L.The weight-average molecular weight and number-average molecular weight of this copolymer was 8 027 and 6 643,respectively,and the multiple-dispersion toefficient was 1.17.The block ratio for PEG and PLGA was 3:2.Drug-loaded PLGA-PEG-PLGA hydrogel could release ricin in vitro for a total of 18 successive days and simultaneously well maintain cytotoxicity and the structural stability of ricin.CONCLUSION:PLGA-PEG-PLGA copolymer is an ideal carrier for sustained delivery of ricin

Objective To explore the effects of low frequency electric stimulation on the clinical manifes- tation and its influence on the level of plasma Endothelin (ET),atrial natriuretic peptide (ANP),thromboxane B_2 (TXB_2),angiotensinⅡ(AngⅡ),Calcitonin Gene Related Peptide(CGRP) and Neuron-specific Enolase(NSE) and its clinical significance in the patients with vertebrobasilar insufficiency(VBI). Methods One hundred and tewenty cases of VBI patients were randomly divided into two groups:a treatment group and a control group.Both groups received routine drug treatment.The treatment group (n=60) was also treated with low frequency electric stimulation at bilateral mastoid processfes in addition.The improved rate of the clinical symptom,the plasma level of ET,ANP,TXB_2,AngⅡ,GRP and NSE were observed after treatment. Results The patients treated in the treatment group had significantly better outcome than the control group in terms of symptom relief.The effective rate was 87.93% and 67.80% in the treatment group and control group,respectively(P