A novel method was developed for the direct analysis of testosterone, androstenedione, methyltestosterone and methenolone in serum samples by fully automated online turbulent flow solid phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry. An aliquot of 50 μL serum sample was preconcentrated directly on a Turboflow SPE column after centrifugation. Turboflow SPE C18-P could be used to remove serum matrix effectively. The optimum loading flow rate and elution time were 4. 0 mL/min and 1. 0 min, respectively. The linearity ranges were from 1. 0 μg/L to 100. 0 μg/L for four target compounds. The method limits of detection ( LODs) were in the range of 0. 2-0. 3 μg/L. The relative standard deviations (RSDs) ranged from 2. 9% to 14. 1% (n=5). The time for one sample analysis including extraction, separation and determination was 32 min. The proposed method has been successfully applied for the analysis of serum samples.

0.05).The expression began to increase around the infarction at 12h reperfusion,it reached the peak at the 3th day in the boundary zone and then it gradually decreased.Conclusion The overexpression of IL 6 played an important role in the course of cerebral ischemic reperfusion, and could damage the brain tissue.

BACKGROUND: Based on the characteristics of cartilage tissue, such as consisting of single type of cells, the cartilage cells or chondrocyte, absence of blood vessel, rather low consumption level of oxygen and nutrition, low level of allo-immunocompetence and simple function in vivo, it seems to be easy for cartilage cell lines to be established for tissue and cell transplantation. We want to set up a cell line with the purpose of current use in tissue engineering in vitro. It will provide the basis for artificial tissue and organ that will become to be standardized and yielded in batch.OBJECTIVE: To explore the potential stimulatory effects of basic fibroblast growth factor(bFGF) and insulin on the proliferation and differentiation in primary culture mice chondrocytes in vitro. The effect and application of the cell factors will be evaluated for tissue engineering.DESIGN: A grouping controlled and repeated trial was conducted with the cells as the subjects.SETTING: Key laboratory of tissue transplantation and immunology of a college.MATERIAIS: The experiment was completed in the Key Laboratory of Tissue Transplantation and Immunology of the Ministry of Education, Jinan University from November 2002 to May 2003. Cultured cartilage cells at random were obtained as the study objects.METHODS: Mice cartilage cells were cultured in medium at the minimum concentrations of serum. The effects of different concentration of bFGF and insulin on the proliferation and differentiation in mice cartilage cells were observed with WST1 and immunofluorescence staining.MAIN OUTCOME MEASURES: Primary results: ① Effect of bFGF on proliferation of primary cultured mice cartilage cells. ② Effect of insulin on proliferation of primary cultured mice cartilage cells. Secondary results:morphological observation of cartilage cells RESULTS: Primary cultured mice cartilage cells were cultured in medium at the minimum concentration of serum(4 g/L fatal bovine serum). It was found that bFGF and insulin might play an important role on the proliferation and growth of mice cartilage cells in a dose-dependent manner. In addition, morphological observation of cartilage cells showed that both bFGF and insulin not only promoted the proliferation of the cells but also enhanced the matrix secretion of cartilage cells.CONCLUSION: Both bFGF and insulin can stimulate the proliferation of cartilage cells in vitro.

Objective To investigate the effectiveness of organ dose modulation (ODM) technology with respect to dose reduction and image quality of the sensitive organs during CTA of craniocervical arteries. Methods Ninety one patients suspected of arterial vascular disease and evaluated by CTA of craniocervical arterires were enrolled in this study. Patients were divided into group A (n=46) and group B (n=45) according to the order of examination. All patients hadCTA examination of craniocervical arteries. The ODM technique was used only in plain phase in group A and was used both in the plain phase and enhanced phase in group B. The tube current in the four direction (anterior,posterior,left,right) of thyroid and orbital area and radiation dose were recorded. The image quality of group A and group B was evaluated by objective indicators [CT attenuation values and contrast-to-noise ratio (CNR) of vessels in the thyroid area and orbital region ]and the subjective score. The independent samples t test was used for the statistical analysis of the objective indicators and nonparametric rank sum Mann-Whitney U test were used for subjective score. Results For A group, There were significant differences for the tube current at all directions of the thyroid gland and orbital area between the plain phase and the enhanced phase except the current at the posterior of the thyroid gland area(P value all<0.05),and the current in plain phase lower than that in enhanced phase. The mean current of thyroid gland and the orbital area were (604±43) mA and (656± 22) mA in the plain phase and (341±54) mA and (409±63) mA in enhanced phase. The differences were significant (t values were -10.909 and -38.454,P<0.01).CTDIvol and ED were (15.6±1.4) mGy and (1.44± 0.17) mSv in the plain phase and (17.4 ± 1.4) mGy and (1.60 ± 0.18) mSv in the enhanced phase, the differences were significant (t values were - 42.008 and - 32.130, P<0.01) . There was no significant differences for the objective indicators and the subjective score between group A and group B(P>0.05). Conclusion ODM technique can provide protective effect on the sensitive organs during craniocervical CTA examination without compromising the image quality.

Objective To investigate the relationship between fetuin A and left ventricular function and their influences on residual renal function(RRF) in peritoneal dialysis patients.Method Eighty patients recently initiating peritoneal dialysis were enrolled into this study and were divided into high fetuin A group and low fetuin A group accordin to the value of serum fetuin A concentration.Hemoglobin,high sensitive C reactive protein(hsCRP),calcium,phosphorus,albumin,lipoproteins and left ventricular myocardial performance index(LV-MPI) were examined.All these patients were followed up for 12 months,to discover the parameters' differences between two groups and to investigate the association between fetuin A and left ventricular function and RRF.Results At the beginning of the study,there was no difference of hsCRP,calcium,phosphorus,albumin,lipoproteins and LV-MPI,estimated glomerular filtration rate (eGFR) between two groups;After 12 months follow-up,MPI was obviously shorter (P ＜ 0.05) and RRF was obviously higher (P ＜ 0.05) in high fetuin A group than thosein low fetuin A group.Compared with the beginning of the study,LV-MPI was significantly increased and eGFR was significantly decreased after 12 months follow-up (both P ＜ 0.05) in low fetuin A group,but no obviously change of LV-MPI or eGFR was found in high fetuin A group after followup.Pearson correlation analysis discovered an obvious negative correlation between fetuin A and MPI (r=-0.680,P ＜ 0.01).Multiple regression analysis indicated that eGFR had positive correlation with fetuin A (B=0.058,t=3.679,P＜ 0.01) and negative correlations with MPI (B=-0.511,t=-2.903,P=0.007),age(B=-0.144,t=-4.013,P＜0.01).Diabetes was risk factor to loss of RRF (B=-2.031,t=-2.759,P ＜ 0.05).Conclusion Fetuin A has very close relationship with left ventricular function.Decreased serum fetuin A level and decreased left ventricular function are risk factors to the loss of the RRF in ERSD patients.

Objective To investigate the association of vascular calcification,fetuin A and Creaction protein (CRP),and explore the influence on cardiovascular events.Methods Sixty peritoneal dialysis (PD) patients were enrolled in this study.Carotid intima-media thickness (cIMT),fetuin A and CRP,along with the other serum related parameters were detected to find out their influence on vascular calcification in PD patients.The relationship between cIMT,fetuin A,CPR and cardiovascular events was analyzed in PD patients with 18 months followed-up.Results Of the 60 PD patients,carotid intima-media thickness (cIMT) was increased in 38 patients(63.3％).Compared with the non-increased cIMT patients,serum fetuin A concentration was significantly decreased(P ＜ 0.05),CRP(P＜0.01) and calcium × phosphate products(P＜0.05) were significantly increased in the highincreased cIMT group.Compared with the low-increased cIMT patients,fetuin A concentration was obviously lower(P ＜ 0.05) and calcium×phosphate products were obviously higher(P ＜ 0.05) in the highincreased cIMT group.Linear regression analysis discovered an obvious negative correlation between CRP and fetuin A(R2 =0.629,F=47.522,P ＜ 0.01),as well as fetuin A and calcium×phosphate products (R2=0.299,F=11.948,P=0.002).Multiple regression analysis indicated that fetuin A was independently negatively correlated with cIMT(B=-0.019,t =-6.042,P ＜ 0.01).At 18 months,there were 36 newly-happened cardiovascular events and among which 6 cases died.Logistic regression analysis found that increased cIMT was risk factor to cardiovascular events in PD patients(OR=3.691,95％CI 1.467-9.258,P=0.006).Conclusion Decreased fetuin A and increased calcium×phosphate products deteriorate carotid calcification in PD patients.Micro-inflammation of PD patients represented by high CRP levels may increase calcium×phosphate products by depressing the fetuin A level,and in the end will stimulate carotid calcification.Increased cIMT is a risk factor for cardiovascular events.

Objective To investigate the therapeutic effects of repeated transcranial magnetic stimulation (rTMS) at high and low frequency for the rigidity and tremors of Parkinson's disease (PD).Methods Sixty-nine patients with PD received rTMS at 5 Hz or 0.5 Hz.The efficacy was assessed using the unified Parkinson's disease rating scale (UPDRS),motor function tests and motor evoked potentials (MEPs).Results Thirty-two patients with rigidity from PD received 5 Hz rTMS therapy,and their UPDRS total scores decreased significantly.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Thirtyseven patients with tremors from PD received 0.5 Hz rTMS therapy,and their UPDRS total scores were significantly decreased.Their motor function test results improved significantly and their MEP relaxed motor thresholds were significantly elevated.Conclusion High frequency rTMS may improve the symptoms of patients with PD rigidity and low frequency rTMS may improve the symptoms of patients with PD tremors.

Objective To examine the expressions of serum growth hormone (ghrelin), leptin (LP), in sulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP3) in children with id-iopathic short stature (ISS), and their significance. Methods A total of 40 patients with ISS were selected from May. 2012 to Oct. 2014 in Fuyang Maternal and Child Health Hospital, while a total of 40 children in good health were selected as the control group. Serum ghrelin and LP levels were measured by ELISA. Serum IGF-1 and IGF-BP3 levels were detected by chemiluminescence immune assay. Results were analyzed statistically. Results The height, weight, BMI, GH, serum LP, IGF-1, and IGFBP3 levels in the observation group were significantly lower than those in the control group, while serum ghrelin expression level was significantly higher than that in the control group(P<0.05). The correlation analysis showed that ghrelin and LP were negatively correlated(r=-0.611, P<0.01), ghrelin and IGF-1 levels were negatively correlated(r=-0.520, P<0.05), Ghrelin and IGFBP3 were pos-itively correlated (r=0.586, P<0.01), IGF-1 and IGFBP3 was negatively correlated (r=-0.576, P<0.01), and LP and IGFBP3 were negatively correlated (r=-0.609, P<0.01). Conclusions It shows that ghrelin, LP, IGF-1, and IGFBP3 levels in children with ISS are related to growth hormone secretion status. The interactions between ghre-lin and insulin-like growth factor axis regulate growth and development of children.

Objective To analyze the influence factors of maintenance dialysis patients with carotid plaque and explore its relationship with cardiovascular disease(CVD). Methods 52 patients on peritoneal dialy-sis and 46 patients on maintenance hemodialysis were involved in this study. Bilateral carotid intima-media thick-ness(IMT)and plaque,the related clinical indicators and CVD were recorded. According to the mode of dialysis, carotid plaque,CVD respectively,we divided our population into two groups,and statistical analyses were made between the two groups. Results Age,percentage of male,the number of hypertension and diabetes,blood uric acid ,blood glucose ,the incidence of cardiovascular disease are higher ,but the level of albumin is lower in the carotid plaque group. Binary Logistic Regression analysis showed that hypertension ,albumin and uric acid signifi-cantly influenced carotid plaque formation (P < 0.05). CVD is closely related to the carotid plaque (r = 0.415 , P = 0.000). Left and right IMT value are higher in the patients with CVD (P < 0.01);carotid plaque mainly distributed in the common carotid artery(CCA)and carotid artery bifurcation(BIF),and with the larger number in the patients with CVD(P<0.01). Conclusions Carotid plaque formation is closely related with the occurrence of CVD in maintenance dialysis patients. Hypertension ,albumin and uric acid are risk factors for the formation of carotid plaque,so dialysis patients in addition to maintain the blood pressure and improve nutrition,are also very important to control the level of uric acid.

Objective To observe the difference of lung injury between toll-like receptor 4 (TLR4) mutant mice and wild type mice in a model of lipopolysaccharide (LPS) induced acute lung injury (ALI),discuss the role of TLR4 in ALI.Methods Different doses of LPS solution (1,5mg?kg~(-1)) were injected in vein tail to reproduce ALI model in both TLR4 mutant (TLR4~(-/-)) and wild type (WT,TLR4~+(/+)) mice.Lung tissues were collected for gross and micrographic histological injury analysis and for assessment of lung edema.Meanwhile,the levels of myeloperoxidase (MPO) in lung tissues in both strains were assessed to evaluate the extent of polymorphological neutrophils (PMN) infiltration.Results The gross and micrographic injury of lung was milder in TLR4 mutant mice than that in wild type mice.The extent of lung edema (W/D) was also reduced compared with wild type mice,especially in 5 mg?kg~(-1) group [(4.08?0.1)vs.(4.55+0.2),n=10,t=12.71,P＜0.01].With high dosage of LPS,the value of W/D in both mice strains was higher than that in sham operation group (P＜0.01).The extent of PMN infiltration in lung tissues in TLR4 mutant mice was reduced compared with wild type mice.But they were higher than sham operated mice (P＜0.01).Conclusion TLR4 May involve in the development of ALI,by sequestration of PMN into lung tissues.