The relationships between cholinergic neurons and SP terminals were examined in the rat sacral ventral horn at the light and electron microscopic levels by means of double immunostaining methods. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase (CHAT) with the avidin-biotin technique and stained bluish-green by indolyl-?-galactoside reaction products with ?-galactosidase as a marker. On the same sections, SP fibers were labeled by polyclonal antisera to SP after application of the peroxidase-antiperoxidase (PAP) method and stained brown by diaminobenzidine (DAB)reaction. At the light microscopic level CHAT-I neurons stained bluish-green and SP- I fibers Stained brown were found in the ventral horn. At the electron microscopic level, many asymmetrical axodendritic synapses(type I of Gray)were observed between CHAT-I dendrites and SP-I terminals in the ventral horn, but axosomatic synapses and symmetrical synapses (type I of Gray) were hardly detected. These results indicate that SP-I terminals make direct synapses with CHAT-I motoneurons of sacral ventral horn. These synapses may be predominantly excitatory and have importance in the control of muscular constriction.

BACKGROUND:Recent investigations show that there are a small part of self-renewing and multi-potent cells named as tumor stem cells.Those cells share many characteristics with somatic and embryonic stem cells and are thought to be responsible for driving tumor progression in growing list of neoplastic diseases.Vascular endothelial growth factor(VEGF)is an important angiogenesis factor,which can regulate endothelial proliferation,angiogenesis,permeability of blood vessel,and thrombogenesis.OBJECTIVE:To summarize the progress of cancer stem cells and tumor angiogenesis.METHODS:A computer-based online search was conducted in PUMMED,CNKI,and Wanfang databases with the key words of "cancer stem cells,vascular endothelial growth factor,new vascularize" in both Chinese and English from January 2000 to October 2009.RESULTS AND CONCLUSION:Among 153 articles,there were 13 in Chinese and 140 in English.Titles and abstracts were preliminarily screened,and articles which were non-relative(n=35),duplicated(n=30),and Meta analysis(n=57),were excluded.A total of 31 articles were included in the final analysis.Angiogenesis regulated by multiple factors was a necessary link for tumor growth and transferring.Stem cells promoted angiogenesis,thereby promoted tumor growth and transferring.However,the modified stem cells caused the opposite effects.On the other hand,the stem cells were considered as a substance vector to localize tumor focus,in particular,it will be potential for looking for subclinical focus and distal focus.

Objective:To construct a eukaryotic expression vector for wild-type human Interleukin-13(whIL-13)and its mutant(mhIL-13) gene fused with enhanced green fluorescent protein(EGFP) and to analyze the expression and subcellular location of the fusion protein in COS-7 cells.Methods:The whole whIL-13 and mhIL-13 coding gene was amplified by RT-PCR. And then the expression vector for EGFP-whIL-13, EGFP-mhIL-13 fusion protein were constructed.The recombinant fusion gene were transfected into COS-7 cells.The expression and subcellular location of the fusion protein were detected by confocal microscopy.Results:The construction of expression vector were all corrected.In the positive cells, green fluorescent was found to be localized in cytoplasm,while empty in nucleus.Conclusion:The expression vectors for EGFP-whIL-13, EGFP-mhIL-13 fusion protein were constructed successfully and expressed in COS-7 cells. Fusion proteins were distributed in cytoplasm and no significant difference was observed between them.

Objective To estimate the influence of 1,25(OH)2D3 on the proliferative ability of and methylation levels of genomic DNA and proliferation-associated gene promoter in human HaCaT keratinocytes.Methods Some cultured HaCaT cells were treated with 1,25 (OH)2D3 of 10-6,10-7 and 10-8 mol/L for 24 hours,then,methyl thiazolyl tetrazolium (MTT) assay was carried out to evaluate the proliferative activity of cells,and a global DNA methylation quantification kit was used to determine the global DNA methylation level.Real-time PCR was conducted to quantify the mRNA expression of DNA methyl transferases (DNMTs) and methyl-DNA binding domain (MBD) proteins,and methylation-specific PCR (MS-PCR) to evaluate the methylation status of promoter region in the programmed cell death 5 (PDCD5) and tissue inhibitor of metalloproteinase 2 (TIMP2) genes,in HaCaT cells after 24-hour treatment with 1,25 (OH)2D3 of 10-6 mol/L.The HaCaT cells receiving no treatment served as the control.Results Compared with the untreated HaCaT cells,those treated with 1,25(OH)2D3 of 10-6 mol/L showed significantly down-regulated proliferative activity (0.152 ± 0.027 vs.0.290 ± 0.017,P ＜ 0.01),global DNA methylation level (0.187 ± 0.071 vs.0.316 ± 0.049,P ＜ 0.05),DNMT3a and DNMT3b mRNA expression levels (P ＜ 0.01 or 0.05),but markedly upregulated mRNA expression levels of MECP2,MBD2,PDCD5 and TIMP2 (P ＜ 0.01 or 0.05).Moreover,the DNA methylation levels within the promoter region of PDCD5 and TIMP2 genes were significantly lower in HaCaT cells treated with 1,25 (OH)2D3 of 10-6 mol/L than in the control cells (0.38 ± 0.135 vs.0.72 ± 0.121,0.46 ± 0.172 vs.0.68 ± 0.133,both P＜ 0.05).Conclusions 1,25(OH)2D3 may down-regulate the global genomic DNA methylation level of,and modulate the expression of DNA methylationmodifying genes in,HaCaT cells.Furthermore,1,25 (OH)2D3 can decrease the promoter methylation levels but induce the overexpression of PDCD5 and TIMP2 genes,and decelerate the proliferation of HaCaT cells.

Objective:Iris-fixed IOL implantation is a new and effective therapy for phakic eyes. Because the patients are young,they know very few about the operation,which mandated delicate pre-and postoperative nursing. This article discussed how to nurse the patients receiving iris-fixed IOL implantation operation in phakic eyes for super high myopia. Methods:Fifty eight eyes of 30 patients with super-high myopia received iris-fixed IOL (Verisyse IOL) implantation. Pre-and postoperative nursing course were summarized. Results:Focused education through preoperative visits,active cooperation in the operation and meticulous postoperative nursery were beneficial for the operation. Significant vision improvement was found after surgery. Conclusion:The planned,step by step,focused nursing course can improve the effects of operation and reduce the postoperative complication.

BACKGROUND:The surgical treatment of thoracolumbar burst fracture combined with spinal cord injury is to relieve mechanical compression on the spinal cord and reconstruct spinal stability through internal fixation.This theory is derived from a lot of animal experiments and important biomechanical principles,but the available clinical data are stil scarce.OBJECTIVE:To investigate the efficacy of posterior decompression and screw internal fixation for the treatment of thoracolumbar fractures combined spinal cord compression,and to evaluate vertebral body height,kyphosis angle and neurological function.METHODS:A retrospective study was performed among 75 patients with thoracolumbar fractures combined spinal cord compression,who were recruited from Department of Orthopedics,the First People's Hospital of Shunde District of Foshan City,from January 2010 to February 2013.They underwent posterior decompression and internal fixation.The preoperative and postoperative kyphosis angle,vertebral height and neurological function were compared.RESULTS AND CONCLUSION:The preoperative kyphotic angle was (25.2±2.1)° and postoperative kyphotic angle was (8.8±2.3)°,with significant differences (P ＜0.05).Preoperative vertebral body height was (58.4±14.2)％and postoperative vertebral body height was (92.3±6.8)％.According to ASIA score,48 cases had slight impairment of neurological function,scores more than 40 points,and 45 of them had completely recovered walking ability,the recovery rate was 94％; the remaining 27 cases had moderate or severe neurological function impairment,scoring less than 40 points,and 15 of them appeared obvious recovery of neurological function,therecovery rate was 56％.Posterior decompression and internal fixation for treatment of thoracolumbar burst fracture combined with spinal cord injury can effectively restore the vertebral height and kyphosis,and improve significantly neurological function.

Objective To investigate left ventricular systolic function with mitral annulus motion velocity(Sa) with pulsed-wave Doppler tissue imaging(PW-DTI) in patients with coronary arteriosclerosis disease(CAD) after percutaneous coronary intervention(PCI).Methods 52 patients with CAD,who were determined by coronary artery angiography,were divided into two groups,PCI group(n=33) and non-PCI group(n=19).They were followed up with PW-DTI and conventional echocardiography before and 5 d,3 months and 6 months after intervention in PCI ones,before and 6 months after angiography in non-PCI ones.In PCI group,the patients were divided two subgroups: EF≥50% and EF=30%～50%.Results The Sa improved significantly in PCI group 6 months after intervention(P<0.001).EF correlated with Sa(r=0.705,r=0.770 pre-and post-intervention respectively,P<0.0001).Conclusion Sa can be used to evaluate left ventricular systolic function of CAD patients.

AIM:To investigate the effects of C1q/TNF related protein 3 (CTRP3) on the insulin sensitivity of insulin resistant 3T3-L1 adipocytes.METHODS: The insulin resistance model of 3T3-L1 adipocytes was induced by palmic acid cultivation.The adipocytes were treated with different concentrations of recombinant CTRP3 protein (10, 50, 250,1 250 μg/L) for 12 h, and for different times (2, 6, 12, 24 h) at the concentration of 250μg/L.The glucose con-sumption was detected by the glucose oxidase method.The glucose transport ratio was measured by 2-deoxidation-[3H]-glucose intake method.The contents of TNF-αand IL-6 in the supernatant were detected by ELISA.The mRNA expression of TNF-α, IL-6 and glucose transporter-4 (GLUT-4) was measured by real-time PCR.The protein expression of GLUT-4 was detected by Western blotting.RESULTS:Compared with normal control ( NC) group, the glucose consumption and glucose intake ratio of insulin resistance ( IR) group was decreased by 50.6%and 57.9%, respectively.Compared with IR group, with the increase in CTRP3 (10, 50, 250,1 250 μg/L) in intervention groups, the glucose consumptions were in-creased by 22.1%, 42.9%, 76.6% and 80.5%, respectively, and the glucose intake ratios were increased by 39.0%, 68.0%, 108.0%and 111.0%, respectively.With the increased duration (2, 6, 12 and 24 h) of CTRP3 treatment at the concentration of 250 μg/L, the glucose intake ratio was increased by 23.0%, 79.0%, 109.0%and 114.0%, respectively. The contents of TNF-αand IL-6 in the supernatant were decreased by 17.4%and 17.1%respectively as treated with CTRP3 at the concentration of 250 μg/L for 12 h, and the mRNA expression of TNF-αand IL-6 was decreased by 26.0% and 18.9%respectively, while the mRNA and protein expression of GLUT-4 was increased by 61.5%and 55.6%respectively. CONCLUSION:CTRP3 may increase the insulin sensitivity of insulin resistant 3T3-L1 adipocytes by down-regulating the expression of inflammatory factors, improving the insulin signal transduction and increasing the expression of GLUT-4.

Objective To investigate the impacts of low-grade inflammation on the correlation of serum testosterone (T) and carotid intima-media thickness (CIMT) in type 2 diabetes mellitus (T2DM) men.Methods Based on the concentration of C-reactive protein (CRP) and T,a total of 247 patients was divided into low-grade inflammation with low T group (LI-LT,CRP ≥ 2.0 mg/L,T ＜ 12.0 nmol/L,n =65),low-grade inflammation with normal T group (LI-NT,n =67),non-low-grade inflammation with low T group (NLI-LT,n =56),and non-low-grade inflammation with normal T group (NLI-NT,n =59).General information,medical history,and anthropometry data were collected.Glycosylated hemoglobin AI c (HbA1c),blood fat,and CIMT were detected.Results Compared to NLI-NT group,CIMT in NLI-LT group was increased without statistical significance [(0.87 ±0.09) vs (0.90 ±0.10)mm,t =1.693,P =0.090].CIMT in LI-LT group was increased significantly compared to that of LI-NT group [(0.99 ± 0.10) vs (1.07 ±0.12)mm,t =5.208,P =0.000].Correlation analysis indicated that serum T correlated negatively with CIMT (n =247,r =-0.368,P ＜0.01) in whole.The correlation coefficient of T and CIMT was-0.582 (P =0.000),and-0.098 (P =0.087) in patients with (n =132) and without (n =115) low-grade inflammation,respectively.To make CIMT dependent coefficient and serum T independent coefficient in multiple regression analysis,the partial regression coefficient was-0.062 (95％ CI:-0.094 ～-0.029,P =0.008),and-0.045 (95％ CI:-0.087 ～-0.002,P =0.036),respectively,before and after the adjustment of age,smoking,family history,T2DM course,body mass,blood pressure,HbAlc,and blood fat.After the additional adjustment of CRP,the partial regression coefficient was-0.019 (95％ CI:-0.120 ～ 0.042,P =0.287).Conclusions The negative relationship between serum T and CIMT in T2DM men might be modulated by low-grade inflammation.

Objective To evaluate global and segmental right ventricular ( RV ) systolic functions in patients with excessive volume or pressure load using real‐time three‐dimensional echocardiography ( RT‐3DE) . Methods Forty‐five patients with RV volume overload ,45 patients with RV pressure overload and 45 healthy subjects were underwent RT‐3DE . RV global and segmental ( inflow ,body ,outflow ) end‐diastolic volume (EDV) ,end‐systolic volume (ESV) ,stroke volume (SV) and ejection fraction (EF) were analyzed with TomTec software . The correlations between EF with the three‐dimensional method and two‐dimensional parameters including right ventricle systolic pressure( RVSP) were discussed . Results Global EDV and ESV increased significantly in both patient groups compared with controls ( all P < 0 .05) ,but there was no difference between two patient groups ( P >0 .05) .Compensated increase of SV was found in sixty percent of patients with volume overload but none with pressure overload ( P < 0 .05) . Global EF decreased significantly in both of patient groups (all P <0 .05) ,which was more significant patients with pressure overload ( P < 0 .05 ) . Different patterns of the regional dysfunction were found among the different RV segments . No correlation was found between RVSP and global or segmental EF in patients with pressure overload . Conclusions RT‐3DE could be used to assess global and segmental RV systolic function in patients with pressure and volume overload .