Objective To explore the dissociation, purity and the culture method of spinal motoneurons in vitro and to find out an effective way to identify spinal motoneurons.Methods Spinal motoneurons were isolated from spinal cord of embryonic rat. Spinal tissues were digested and then the motoneurons through 6.8% metrizamide density gradient centrifugation, were obtained by collecting the top layer of metrizamide cushion. After the glia adhered to the plate wall, the enriched motoneurons were collected and plated in 24-well plates at a density of 4?10~5/ml. Cytosine arabinoside was added to the culture system after 24 hours in order to inhibit the outgrowth of glia. L-15 serum medium was used for the first 48 hours and then changed by serum-free medium. Later, neurons were cultured with 50% exchanged medium every two days. Spinal motoneurons were identified by immunostaining with polyclonal antibody to choline acetyltransferase (ChAT) according to ABC method. Laser scanning confocal microscope was used to observe its structure.Results Spinal motoneurons were immunoreactive to the antibody against ChAT. The present experiment revealed that ChAT-positive cells more than 85% might live about seven to nine days.Conclusions This study suggests a method of spinal motoneuron culture successful. Spinal motoneurons could be specially labeled with ChAT antibody.

Taking the Wechat public service platform of Administration of Traditional Chinese Medicine of Yunnan- Yunnan Zhongyi as an example,the paper uses the Importance-performance Analysis (IPA) and corresponding analysis method,from the users' satisfaction perspective,evaluates 28 satisfaction indicators,discusses the relationship between the importance recognition of health Wechat of Traditional Chinese Medicine (TCM) and user characteristic,so as to propose the development of individual TCM health information recommendation and strengthen the effective interaction of platform users.

Aim To investigate the expression of G protein-coupled receptor TGR5 and its effects on FN and TGF-β1 expression cultured under high glucose condition in rat glomerular mesangial cells , and then to explore the role of TGR5 in diabetic nephropathy. Methods INT-777 and TGR5 plasmid were used to activate TGR5 under high glucose(HG,30 mmol·L - 1 glucose ) condition, and anti-TGR5 small interfering RNA(TGR5 siRNA) was used to knock down TGR5. The protein expression of FN and TGF-β1 in rat me-sangial cells was detected by Western blot. Results TGR5 could be detected in rat glomerular mesangial cells. Both FN and TGF-β1 protein levels could be in-creased by high glucose compared with control group(P < 0. 05),and be inhibited by activiation of TGR5(P <0. 05). On the other hand,knockdown of TGR5 could increase FN and TGF-β1 protein to abnormal levels(P< 0. 01,P < 0. 05). Conclusion TGR5 suppresses HG-induced FN and TGF-β1 expression in rat glomer-ular mesangial cells,suggesting a protective role in the process of diabetic nephropathy.

Objective To explore the roles of p53 in ionizing radiation induced MCF-7 cell cycle uncoupling. Methods The p53 knock-down models was established in MCF-7 with retrovirus packaged particles from 293T cells through calcium acid phosphate co-precipitation, then Western blot was used to detect the protein expression. Flow cytometry(FCM) was used to analyze the cell cycle uncoupling and polyploid after irradiation. Results Compared with p53+/+ group, the percentages of G0/G1 cells in p53 -/- group decreased, while those of S and G2+M increased (P < 0.01). In polyploidy analysis 2N cells decreased, whereas both 4N and 8N cells =increased (P<0.01). Compared with sham-irradiation, 4 Gy X-ray led to the decrease of G0/G1, S cells, and the increase of G2+M cells. The increase of 2N cells and decrease of 4N and 8N cells were observed in both p53+/+ and p53-/- cells. Compared with p53+/+ +IR group, the decrease of G0/G1 and S cells and the increase of C2 + M cells were significant (P < 0.01) in p53-/-+ IR groups. 2N cells decreased, 4N cells increased, but no changes in 8 N cells occurred. Conclusion Radiation might induce G2 arrest and cycle uncoupling, p53 plays a role in the regulation of G2 arrest, but no role in cycle uncoupling.

Objective To evaluate the roles of multi-voxel proton magnetic resonance spectroscopy (1H-MRS) in the early diagnosis of vascular cognitive impairment no dementia (VCIND).Methods Seventy-eight out-patients and inpatients in Department of Neurology, the second Affiliated Hospital of Soochow University from December 2008 to September 2009 were recruited in this case-control study.Their cognitive functions were assessed with a wide range of neuropsychological battery of tests including Montreal cognitive assessment ( MoCA Beijing Version ), auditory verbal learning test ( AVLT), digital span test (DST), Rey-Osterrieth complex figure test (CFT) ,semantic and phonetic fluency tests, digit symbol coding subtest (DSCT), trail making test (TMT), clock drawing test (CDT) and the Stroop color-word test (SCWT).All patients were classified into vascular cognitive impairment no dementia (VCIND) group and cognitive normal control group based on the results of neuropsychological tests.Eighteen patients with VCIND and 18 gender-, age- and education-matched normal control were randomly selected for the following study.With multi-voxel 1H-MRS, the levels of N-acetylaspartartate (NAA), creatine (Cr) and choline (Cho) in gray matters of bilateral frontal lobe, temporal lobe, parietal lobe and thalamus were measured and the ratios of NAA/Cr, Cho/Cr were compared between the two groups.Meanwhile, the correlations between scores of MoCA and its sub-items and the ratios of NAA/Cr, Cho/Cr were analyzed in VCIND group.Informed consent was obtained from all participants and the study was approved by the Ethics Committee of the hospital.Results Compared with control group, the ratios of NAA/Cr were significantly decreased in bilateral gray matters of thalamus ( left, 1.56 ± 0.49 vs 1.89 ± 0.48, F = 11.222, P = 0.002; right,1.63± 0.45 vs 1.86 ± 0.33, F = 5.358, P = 0.027 ).No significant difference were found in NAA/Cr in gray matters of bilateral frontal lobe, temporal lobe, parietal lobe and Cho/Cr in all regions between two groups ( all P ＞ 0.05 ).In VCIND group, the decreased degree of NAA/Cr in bilateral gray matters of thalamus was significantly positively correlated with the MoCA total score ( r = 0.54, 0.44 ) as well as the sub-scores in tested items of memory ( r = 0.61, 0.49 ), attention ( r = 0.43, 0.36 ), language ( r = 0.39,0.31) and visuospatial or executive( r = 0.29 , 0.33, all P＜0.05 orP＜0.01).Conclusions Cognitive impairment in patients with VCIND maybe related to metabolic dysfunction of neurons in bilateral thalamic.Multi-voxel 1H-MRS plays an important role in early diagnosis and monitoring disease progression of VCIND.

Massive hemoptysis is one of the most dreaded of all respiratory emergencies and can have a variety of underlying causes. It is mostly caused by bleeding from bronchial circulation. Bronchial artery embolization is now considered to be the treatment of choice for acute massive hemoptysis. Bronchial artery embolization (BAE) is a safe and effective nonsurgical treatment for patients with massive hemoptysis. However, nonbronchial systemic arteries can be a significant source of massive hemoptysis and a cause of recurrence after successful BAE. So knowledge of the bronchial artery anatomy, together with an understanding of the pathophysiologic features of massive hemoptysis, are essential for planning and performing BAE in affected patients. In addition, interventional radiologists should be familiar with the techniques, results, efficacy, safety and possible complications of BAE and with the characteristics of the various embolic agents. Bronchial arterial catheterisation in human via a percutaneous approach has been practiced for 32 years (1973) in the world and 20 years (1986) in China, initially for direct chemotherapy treatment for bronchial malignancies and then for the embolization of patients with massive haemoptysis. A review of clinical experience to evaluate technique,embolic materials,outcome and complications of BAE is presented.

Objective To investigate the diagnostic value of diffusion kurtosis imaging(DKI)in the classification of hepatic fibrosis. Methods Thirty-five male SD rats were randomly divided into two groups:the hepatic fibrosis group(n=28)and the control group(n=7). The rats in hepatic fibrosis group were randomly divided into 4 subgroups and seven rats per group, the rats were administrated 50％ CCl4 intraperitoneally twice a week to establish hepatic fibrosis , and the four subgroups were injected 2, 4, 6, and 8 weeks, respectively. The rats in the control group were administrated same dose of olive oil for 8 weeks. One rat in hepatic fibrosis group was died of liver failure in the 7th week, and a total of 27 fibrosis experimental rats and 7 control rats were finally included in this study. DKI was performed at the end of the injection period for all rats, the apparent diffusion(D)and kurtosis(K)values were evaluated. Rats were sacrificed immediately after MRI scan and liver specimens were collected. The liver tissues were examined by pathology, liver fibrosis degree, which was graded from S0 to S4, and inflammatory activity, which was graded from G0 to G3 were graded. The difference of D value and K value between different liver fibrosis and inflammatory activity scores was compared by one-way ANOVA(normal distribution)or Kruskal-Wallis test(skewed distribution). Spearman correlation analysis and multiple regression analysis were used to reveal the correlation between DKI parameters and fibrosis staging/necroinflammatory activity grade. To confirm the efficiency of using the ROC curve of DKI parameters to qualify the liver fibrosis grade, which grade was≥3. Results Seven, 6, 6, 7, 8 rats were diagnosed as S0 to S4, respectively. The difference of D value and K value among different fibrosis grades was statistically significant(P<0.05). D value and the degree of fibrosis was negatively correlated(r=-0.650, P<0.01);K value and liver fibrosis grade no correlation(r=0.336, P=0.080). Thirteen, 6, 8, 7 rats were diagnosed as G0 to G3, respectively. D value was negatively correlated with inflammatory activity(r=-0.590, P=0.001);K value was no correlation with inflammatory activity(r=0.169, P=0.389). Compared with inflammatory activity, fibrosis classification was an independent factor in determining D values(P=0.001). ROC analyses demonstrated an area under the curve(AUC)of D value, K value, D value combined with K value in the diagnosis of liver fibrosis grading ≥ 3 level were 0.781, 0.672 and 0.833, respectlively. The sensitivity and specificity of D value combined with K value were 83.3％ and 75.0％, respectively. Conclusion DKI imaging is of great value in the classification of hepatic fibrosis and can be used as an effective method for the diagnosis of fibrosis.

Objective To evaluate dual-source dual-energy CT(DSCT) for the differentiation of urinary stone composition in vitro. Methods Ninety-seven urinary stones were obtained by endoscopic lithotripsy and scanned using dual-source dual-energy CT. The stones were divided into six groups according to infrared spectroscopy stone analysis: uric acid ( UA ) stones ( n = 10 ), cystine stones ( n = 5 ), struvite stones( n = 6), calcium oxalate ( CaOx ) stones ( n = 22 ), mixed UA stones ( n=7 ) and mixed calcium stones(n=47). Hounsfield units (HU) of each stone were recorded for the 80 kV and the 140 kV datasets by hand-drawing method. HU difference, HU ratio and dual energy index ( DEI ) were calculated and compared among the stone groups with one-way ANOVA. Using dual energy software to determine the composition of all stones, results were compared to infrared spectroscopy analysis. Results There were statistical differences in HU difference [(-17±13), (229±34),(309 ±45), (512 ±97), (201±64)and (530±71) HU respectively], in HU ratio (0.96±0.03, 1.34 ±0.04, 1.41 ±0.03, 1.47 ±0.03,1.30±0.07, and 1.49 ±0.03 respectively), and DEI( -0.006 ±0.004, 0.064 ±0.007, 0.080 ±0. 007, 0. 108±0.011 ,0. 055 ±0.014 and 0. 112 ±0.008 respectively ) among different stone groups(F=124. 894,407.028, 322. 864 respectively, P ＜0. 01 ). There were statistical differences in HU difference,HU ratio and DE1 between UA stones and the other groups( P ＜ 0. 01 ). There were statistical differences in HU difference, HU ratio and DEI between CaOx or mixed calcium stones and the other four groups (P＜0. 01 ). There was statistical difference in HU ratio between cystine and struvite stones ( P ＜ 0. 01 ). There were statistical differences in HU difference, HU ratio and DEI between struvite and mixed UA stones (P＜0. 05 ). Dual energy software correctly characterized 10 UA stones, 4 cystine stones, 22 CaOx stones and 6 mixed UA stones. Two struvite stones were considered to contain cystine. One cystine stone, 1 mixed UA stone, 4 struvite stones and 47 mixed calcium stones were considered to contain oxalate. Conclusions DSCT has the ability to differentiate urinary stone composition in vitro. With dual energy software, the UA, cystine and mixed UA stones can be differentiated from other types of stones.

Objective To compare the efficacy of add-on adefovir dipivoxil (ADV) therapy and switch-to entecavir (ETV) monotherapy in chronic hepatitis B (CHB) patients with suboptimal response to lamivudine (LAM).Methods A prospective study was performed in 120 CHB patients from Zhuji People' s Hospital and the First Affiliated Hospital of Zhejiang University School of Medicine during June 2010 and June 2011.All patients previously received more than 24 weeks LAM treatment,but HBV DNA was still positive.Patients were randomized assigned to two groups:60 patients received add-on ADV therapy and another 60 switched to ETV monotherapy.Both groups were treated for 48 weeks.Liver and kidney function,alpha-fetal protein (AFP),HBV serum markers,HBV DNA and prothrombin time (PT) were examined,and ultrasonography or CT scan of liver was performed every 1-3 months.x2 test was used to compare the HBV DNA negative rates,HBeAg seroconversion rates,resistance rates and adverse reaction at week 48 between two groups.Results Thirty-three out of 38 patients (86.8％) with baseline HBV DNA 103-105 copies/mL became HBV DNA negative after add-on ADV treatment for 48 weeks,twenty-seven out of 39 patients (69.2％) with baseline HBV DNA 103-105 copies/ml became HBV DNA negative after switch-to ETV treatment.There was a statistical difference between two groups (x2 =4.578,P ＜ 0.05).Sixteen out of 22 patients (72.7％) with baseline HBV DNA ＞ 105 copies/mL became HBV DNA negative after add-on ADV treatment for 48 weeks,while only 52.4％ (11/21) patients achieved HBV DNA negative in the switch-to ETV group.There was also a statistical difference between two groups (x2 =4.865,P ＜0.05).None of patients in add-on group developed virological breakthrough and resistance,while 5 patients in switch-to ETV group developed virogical breakthrough and 3 patients developed genetic mutation.Among them,rtM204V + rtL180M + rtS202G mutation was detected in 2 patients,and rtM204V + rtL180M +rtT184A mutation was detected in 1 patient; all mutations happened in the baseline HBV DNA ＞ 105 copies/mL group.Conclusion The add-on ADV therapy is better in viral inhibition than switch-to ETV therapy for CHB patients with suboptimal response to LAM,and it can reduce the occurrence of drug resistance.

Objective To survey the anxiety status of university students in Xuzhou city ,and to analyze its influence factors to propose the effective improvement strategy .Methods The stratified cluster random sampling method was adopted to conduct the questionnaire survey on 1931 college students from 2 colleges in Xuzhou City .The statistical analysis was performed by using SPSS 16 .0 .Results The average score of anxiety in college students was (42 .23 ± 9 .70) points ,the total detection rate was 21 .4% .The main influencing factors of anxiety included the home ranking ,character ,specialty ,school record ,getting scholarship ,plan to partici-pate in graduate entrance examination ,employment prospect ,sleep quality ,physical condition ,relationship with classmates and ro-ommates ,love status ,work-study programs or go out to work situation ,family type ,communication with parents ,family income . Conclusion The anxiety status of college students in Xuzhou City is in middle level .So improving the college students′anxiety sta-tus needs the joint efforts of school ,family and students themselves .