Objective To estimate the effect of leptin antibody on transforming growth factor (TGF)-β1 mRNA expression in hypertrophic scar model in rabbit ears.Methods Fifteen New Zealand white rabbits were included in this study.Three circular incisions which measured 7 mm in diameter and reached the perichondrium,were made in each ear of these rabbits to establish 90 models of hypertrophic scar.After the operation,these models were randomly and equally divided into 3 groups to be treated with topical sodium chloride physiological solution for 40 days (saline group),topical leptin antibody of 2 ng/ml for 40 days (leptin antibody group),and topical leptin antibody of 2 ng/ml for 14 days followed by injection of leptin antibody of 2 ng/ml once a week for 3 weeks (combination group).Scar tissue was resected from these rabbit ears at 40 days after the operation,followed by the determination of scar elevation index,histopathological examination by using haematoxylin and eosin staining,and quantification of TGF-β31 mRNA expression by real-time fluorescence-based PCR.SPSS 13.0 software was used for data processing.Statistical analysis was carried out by one-way analysis of variance.Results A significant decrease was observed in scar elevation index (2.33 ± 0.33 and 2.35 ± 0.22 vs.3.33 ± 0.41,both P ＜0.05) and TGF-β1 mRNA expression in the leptin antibody group and combination group compared with the control group,whereas no significant difference was observed between the leptin antibody group and combination group in either of the two parameters.Pathologically,there was an apparent proliferation of capillaries in the saline group with numerous irregularly and densely arranged fibroblasts with large nuclei,while relatively few fibroblasts with small nuclei,which were arranged in a more regular way,were observed in the leptin antibody group and combination group.Conclusion Leptin antibody treatment can reduce the expression of TGF-β1 mRNA in hypertrophic scar tissue in rabbit ears.

Objective To study the effect of prodnisolone on lipopolysaccharide(LPS)-induced RANTES by microglia.Methods The microglia were incubated in MEM culture medium,and different concentrations of LPS 0,100,500,1000?g/L) or LPS(100?g/L) and prodnisolone(10?mol/L) were added into the culture medium.12h later,the amount of RANTES was measured by ELISA.Results Microglia markedly produced RANTES in response to LPS.Prodnisolone significantly reduced the production of RANTES in microglia.Conclusion Prodnisolone has an inhibitive effect on the production of chemokines such as RANTES.

Objective To investigate the status of vitamin(Vit)B12 deficiency in elderly inpatients in department of neurology.Methods 827 patients admitted to the department of neurology of Shanghai Punan hospital from March 2007 to July 2008,aged 60 and older,with no severe hepatosis and renal disfunction,and without using VitB12 preparation for 3 months,were included in the study.The levels of serum VitB12,folate,homocysteine(Hcy) were tested to screened patients with VitB12 deficiency.The results of symptoms,positive signs of neurological system and the laboratory examination were analysed and compared between patients with or without VitB12 deficiency.Results There were 827 patients,with the mean age of(77.1?7.5)years old,fulfilled the enrolling criteria.VitB12 deficiency was found in 163 patients(19.7%).The prevalence was higher in female(21.9%)than male(17.4%),which had a tendency of increased with age.In the patients with VitB12 deficiency,the rate of complicating gastrointestinal diseases was higher.Compared with patients with normal serum VitB12 level,there were more cases suffered from unstable gait(27.0%),hypopallesthesia(34.4%),and more cases combined with some chronic diseases such as cerebral infarction(62.6%),hypertension(68.7%),coronary heart disease(39.9%).77% cases had neuroelectricphysiological abnormalities,but only 9.82% cases had megaloblastic anemia.Conclusions VitB12 deficiency is common in elderly inpatients in neurology department,whose clinical manifestations are various and atypical,and the neurological symptoms are more common than megaloblastic anemia.

Objective:To evaluate the impact of health education intervention on promoting rural residents to join new rural cooperative medical system(NCMS)and their intention of joining NCMS based on Health Belief Model in project areas in Henan and Jilin Provinces.Methods:Quasi-experiment study was used to evaluate intervention impact.Following the evidence-based approach,according to needs assessment,a half-year health education intervention was implemented among farmers in the experimental counties in Henan and Jilin Provinces respectively.A questionnaire survey was conducted among farmers in intervention and control counties before and after intervention,and intervention impact was evaluated by comparing the indicators' changes in intervention and control counties.Results:After health education intervention,the knowledge level of farmers in two intervention counties increased by 29.0% and 37.8% respectively,their scores of perceived threatens of health risk and perceived barriers of joining NCMS among the respondents were decreased.Meanwhile,their score of perceived benefit of joining NCMS were increased,and the rate of willingness to join NCMS increased remarkably in both intervention counties.Conclusion:Health education was effective and helpful in increasing farmer's knowledge,understanding and cognitive level of NCMS,and it should play an important role for the sustainable development of NCMS.

Objective To evaluate the diagnostic value of the combination of serum DKK1 (Dickkopf-1 )and P53 autoantibodies in patients with esophageal squamous cell carcinoma (ESCC).Methods Serum levels of DKK1 and P53 autoantibodies were measured by enzyme-linked immunosorbent assay (ELISA) for the 1 26 patients with ESCC and 60 normal controls.Receiver operating characteristics (ROC)was used to calculate the diagnostic efficiency.Results The serum levels of DKK1 and P53 autoantibodies were signifi-cantly higher in ESCC than those in normal controls [(673.09 ±343.82)pg/ml vs (362.05 ±1 48.07)pg/ml, Z =6.1 58,P <0.000 1 ;(0.398 ±0.546)vs (0.069 ±0.050),Z =3.832,P <0.000 1 ].ROC curves showed the optimum diagnostic cutoff for serum DKK1 was 588.77 pg/ml,with an area under curve (AUC)of 0.780 (95%CI:0.71 5 ~0.844,61 .9% sensitivity,95.0% specificity).Measurement of P53 autoantibodies demonstrated an AUC of 0.674 (95%CI:0.598 ~0.750,45.3% sensitivity,95.0% specificity).The com-bination of DKK1 and P53 autoantibodies yielded an AUC of 0.843 (95%CI:0.788 ~0.897,73.8% sensitiv-ity,95.0% specificity).In early-stage ESCC,combined detection of DKK1 and P53 autoantibodies improved the diagnostic power,with an AUC of 0.903 (95%CI:0.845 ~0.961 ,81 .0% sensitivity,95.0% specifici-ty).Conclusion Serum DKK1 and P53 autoantibodies can be used as potential diagnostic biomarkers for the ESCC.Combined detection of them might aid the early diagnosis of ESCC.

Objective:To analyze the influential factors, trend and equity of preventive care utilization in China from 1991 to 2011. Methods:Using China Health and Nutrition Survey ( CHNS) , logistic regression was applied to explore influential factors, study time trend of preventive care utilization, and examine its equity by Concentration In-dex. Results: Total preventive care services and physical examination utilization significantly increased. Significant inequity still exit in terms of general physical examination and blood text, however, disappeared with respect to blood pressure screening and gynecological examination. Main influential factors of preventive care utilization includes in-come, gender, education and insurance, etc. Conclusion:Preventive care utilization had significantly grown with de-creasing inequalities, but inequity still exit, particularly for general physical examination and blood test.

Objective: To explore the effects of N-(4-hydroxyphenyl) retinamide (4HPR), 4HPR liposome (4HPR-L), and 4HPR lipid microbubble (4HPR-LM) combined with ultrasound on proliferation, apoptosis, and cell cycle of human keloid fibroblasts (Fbs). Methods: (1) 4HPR-L and 4HPR-LM were prepared by hydration ultrasonic method. The appearance morphology, particle size distribution, Zeta potential, loading drug concentration, encapsulation efficiency, and drug loading rate of 4HPR-L were investigated by high performance liquid chromatography, dynamic light scattering, and transmission electron microscope. (2) Human keloid Fbs were cultured and divided into 13 groups by random number table (the same grouping method below), with 6 wells in each group. Cells in control group were given no treatment, while cells in 12 ultrasound groups including 0.5 W 30 s group, 0.5 W 60 s group, 0.5 W 120 s group, 0.7 W 30 s group, 0.7 W 60 s group, 0.7 W 120 s group, 1.0 W 30 s group, 1.0 W 60 s group, 1.0 W 120 s group, 1.5 W 30 s group, 1.5 W 60 s group, and 1.5 W 120 s group were treated by ultrasound with corresponding parameters. The cells viability was measured by a microplate reader after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 5 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups were treated with blank lipid microbubbles in corresponding mass concentration. The cells viability was measured as before after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 6 groups, with 12 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, 50, and 100 μg/mL 4HPR-L groups were added with 4HPR-L carrying corresponding mass concentration of 4HPR. The cells viability in 6 wells of each group was detected after 24 and 48 hours of routine culture, respectively. Another batch of human keloid Fbs were divided into 4 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 4HPR, 4HPR-L, and 4HPR-LM+ ultrasound groups were treated with 4HPR, 4HPR-L, and 4HPR-LM (all the mass concentration of 4HPR was 20 μg/mL), respectively, and cells in 4HPR-LM+ ultrasound group were given 0.5 W 60 s ultrasound treatment immediately after drug administration. The cells viability was measured as before after 24 hours of routine culture. (3) Another batch of human keloid Fbs were divided into control group, 4HPR group, 4HPR-L group and 4HPR-LM+ ultrasound group, with 3 wells in each group, and the cells in each group were treated as before. Apoptosis of the cells was detected by flow cytometer after 24 hours of routine culture. (4) Another batch of human keloid Fbs were grouped and treated as in (3), and then the cell cycle distribution was detected by flow cytometer after 24 hours of routine culture. Data were processed with one-way analysis of variance and t test. Results: (1) 4HPR-L particles had a spherical or spheroidal structure and were uniform in size, with particle size of (100.1±1.3) nm and Zeta potential of (-34.3±2.3) mV. The mass concentration of 4HPR in 4HPR-L solution was about 1 400 μg/mL, with the encapsulation efficiency of (95.8±1.2)% and drug loading rate of (8.3±0.4)%. (2) The viability of cells in the 12 ultrasound groups was higher than 93.0%, and the viability of cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups was higher than 95.0%. The viability of cells in 1 μg/mL 4HPR-L group at administration hour 24 was similar to that at 48 (t=0.393, P>0.05). The viability of cells in 10, 20, 50, and 100 μg/mL 4HPR-L groups at administration hour 24 was significantly higher than that at administration hour 48 (t=44.593, 22.961, 32.224, 35.337, P<0.01). The viability of cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group was (47.3±0.7)%, (42.3±1.7)%, and (38.6±0.8)%, respectively. The viability of cells in 4HPR group was significantly higher than that in 4HPR-L group and 4HPR-LM+ ultrasound group (t=4.551, 15.895, P<0.05 or P<0.01). The viability of cells in 4HPR-L group was significantly higher than that in 4HPR-LM+ ultrasound group (t=-3.360, P<0.05). (3) The percentages of total apoptotic cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group were (32.8±2.4)%, (42.5±2.4)%, and (58.5±6.3)%, respectively, which were significantly higher than the percentage of control group [(14.9±1.6)%, t=8.748, 13.637, 9.500, P<0.01]. The percentages of total apoptotic cells in 4HPR-L group and 4HPR-LM+ ultrasound group were significantly higher than the percentage in 4HPR group (t=4.049, 5.393, P<0.05 or P<0.01), and the percentage of total apoptotic cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group (t=3.371, P<0.01). (4) The percentage of G2/M phase cells in 4HPR group was higher than that in control group, but there was no statistically significant difference (t=2.107, P>0.05). The percentage of G2/M phase cells in 4HPR-L group was significantly higher than that in 4HPR group or control group (t=18.169, 30.026, P<0.01). The percentage of G2/M phase cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group, 4HPR group, and control group (t=4.932, 25.854, 66.231, P<0.01). Conclusions 4HPR can inhibit proliferation, induce apoptosis, and arrest G2/M phase of human keloid Fbs, and the effects of 4HPR-LM combined with ultrasound are better than those of 4HPR-L and free 4HPR.

Objective To investigate the wall shear stress(WSS) in the common carotid artery of type 2 diabetes mellitus(DM) patients,and analyzed the spatial distribution of WSS by using quantitative visualization of blood flow shear stress analysis software.Methods Eighteen male type 2 DM subjects were enrolled as DM group and 18 age-matched healthy subjects were selected as control group.None of the participants was hypertensive,hyperlipidemic or a cigarette smoker.Intimal-medial thickness (IMT),number and size of plaques in the common carotid artery were evaluated by high-resolution echo-Doppler.Color Doppler flow images of common carotid arteries in the two groups were extracted from DICOM files.WSS in the common carotid arteries was calculated by shear stress visualization quantitative analysis software,and the corresponding spatial distribution maps of WSS were designed.Results WSS of the common carotid arteries in the control group were ranged from 4 to 14 dyne/cm2.WSS of the common carotid arteries in the DM group were ranged from 2 to 8 dyne/cm2.Compared to mean WSS value [(6.96 ± 1.17)dyne/cm2] of common carotid arteries in the control group,mean WSS value [(3.14 ± 0.79)dyne/cm2] of common carotid arteries in the DM group was significantly lower (t =9.380,P =0.000).Six diabetic participants had a plaque in one carotid artery and no lesions in the contralateral carotid.Among these subjects,mean WSS was significantly lower in the side with lesion (t =7.324,P =0.000).Therefore,IMT of common carotid arteries in participants was significantly inversely related to WSS (r =-0.76,P ＜0.01).Conclusions The common carotid arteries of DM patients are more prone to atherosclerosis which is associated with reduction of WSS.The hemodynamic profile might represent an additional factor contributing to the increased prevalence and severity of carotid atherosclerosis in diabetic patients compared with general population.

Objective To establish a simple and efficient method for developing a keloid model in nude mice with human keloid-derived fibroblasts.Methods Twenty-seven female BALB/c nude mice were randomly divided into five groups with 5,5,5,8 and 4 mice in group A,B,C,D and E respectively.The mice in group A,B and C were inoculated with 0.1 ml of suspension containing human keloid-derived fibroblasts at concentrations of 1.0 × 104,3.0 × 104 and 5.0 × 104 per microliter Matrigel,respectively,at the right axillary fossa.The tumors that formed in one mouse in group C were taken out,and cut into several parts measuring 5 mm × 5 mm × 5 mm in size,which were then subcutaneously transplanted into the right axillary fossa of mice in group D.The mice in group E were subcutaneously injected with 100 μl of Matrigel and served as the control group.The formation of tumor in mice was observed by naked eyes,and the size of tumors was measured until day 30 after tumor formation in group A,B and C as well as after tumor transplantation in group D.Mice were sacrificed on day 30 after tumor formation,and histopathologic examination was performed to analyze histological features of transplanted tumors and pathological changes in visceral organs such as heart,liver,spleen,lung and kidney.Results The tumor formation rate was consistently 100％ in group A,B and C,and the time required for tumor formation was (90.20 ± 3.96),(61.00 ± 2.92) and (39.60 ± 3.20) days in group A,B and C respectively.There was a significant difference in tumor volume on the 30th day after tumor formation between group A,B and C ((288.34 ± 25.29) vs.(1 370.63 ± 105.24) vs.(1 940.98 ± 184.37) mm3,F =138.74,P ＜ 0.05).The size of implanted tumor mass in group D firstly increased,then gradually decreased,but began to continuously increase since the 14~ day,and tumor finally formed in 7 out of 8 mice.There was no evidence of tumor formation in group E.Histopathologic examination showed uniform histological manifestations,which were similar to those of human scar,in tumor tissues from mice in group A,B,C and D.Neither pathological changes nor metastases were observed in visceral organs of these mice.Conclusion Keloid-bearing nude mouse model can be established by subcutaneous inoculation with human keloidderived fibroblasts,or by subcutaneous transplantation of tumor masses of a certain size that have formed in nude mice.

Objective To investigate the effect of intramuscular and oral mecobalamin on hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.Methods One hundred and twenty-six patients with vitamin B12 deficiency who fulfilled the inclusion criteria were randomly divided into 3 groups,42 cases in each,including the intramuscular group (to receive mecobalamin therapy intramuscularly),the oral group ( to receive mecobalamin therapy orally) and the control group ( without mecobalamin therapy).The changes of hematologic markers including hemoglobin (Hb,g/L),mean corpuscular volume (MCV,fl),serum levels of vitamin B12 (ng/L),folate and total homocysteine (Hcy,μmol/L),and neurological signs before and after treatment were compared among these groups.Results Baseline characteristics among the three groups were similar.After a 6-month therapy,there were no differences in any markers in control group patients in comparison to baseline;for patients in the intramuscular group,the blood vitamin B12 levels increased from (139.13 ± 31.57)ng/L to (328.10 ± 42.35 )ng/L (P ＜ 0.001 ).Hcy levels decreased from (36.29 ± 16.23 )μmol/L to ( 18.23 ± 9.85 ) μmol/L ( P ＜ 0.001 ).Hb rose from ( 125.34 ± 16.21 ) g/L to ( 132.79 ± 15.98 )g/L (P =0.037).MCV reduced from (92.98 ±5.35)fl to (87.65 ±5.74)fl (P ＜0.001 ) ;For patients in the oral administration group,the blood vitamin B12 levels increased from ( 138.19 ± 29.95) ng/L to (487.79 ±32.21 ) ng/L ( P ＜ 0.001 ).Hcy levels decreased from ( 33.27 ± 11.51 ) μmol/L to ( 17.49 ± 10.13 ) μmol/L( P ＜ 0.001 ).Hb rose from ( 125.89 ± 17.65 ) g/L to ( 133.46 ± 16.26) g/L ( P =0.041 ).MCV reduced from (93.08 ± 5.10 ) fl to ( 89.29 ± 5.37 ) fl ( P =0.001 ).After the 6-month therapy,there were somewhat improvement in MMSE scores of the intramuscular ( 28.24 ± 3.89 vs.27.85 ± 3.56,P =0.633 ) and the oral groups (27.97 ± 3.77 vs.27.34 ± 3.15,P =0.408 ) compared with baseline,but the differences were not significant.The achilles tendon reflex of the intramuscular ( 1.86 ± 0.67 vs.1.56 ± 0.61,P =0.035 ) and the oral groups ( 1.79 ± 0.64 vs.1.43 ± 0.51,P =0.006 ) were enhanced compared with baseline.Foot vibration sensation of the intramuscular ( 1.35 ± 0.37 vs.1.06 ± 0.41,P =0.001 ) and the oral groups ( 1.24 ± 0.52vs.1.01 ± 0.43,P =0.03) were improved compared with baseline.After treatment,the serum VitB12 concentration in the oral group were higher than that of the intramuscular group ( P ＜ 0.001 ).There were no significant differences in other indexes between the two groups.Conclusion Mecobalamin,administered either intramuscularly or orally,may improve the hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.