Objective To observe the influence of tumor necrosis factor-alpha(TNFα) on skeletal muscle protein catabolism in rats with chronic obstructive pulmonary disease (COPD) and the effects of indomethacin (IND) on it. Methods Duplicated COPD model rats were divided into two groups: the malnutrition group and the normal nutrition group. The malnutrition group were further divided by randomized block design into four groups. Isotonic physiologic saline was administered to group A, the control and the normal nutrition group, and different doses of oral IND were administered to groups B, C, and D weight, concentrations of TNFα, contents of 3-methyl-histidine ( 3- M H ) and tyrosine (Tyr) in the diaphragm and extensor digitorum longus muscle homogenates were measured before and after the intervention. Results Before the intervention, the concentrations of TNFα in the serum of malnutrition groups were all significantly higher than those of normal nutrition group and the control group. After the intervention: (1) The concentrations of TNFα in the serum of the rats of group B, C and D were significantly lower than the group A, especially in group C. The levels of TNFα in serum and body weight of model group rats were negatively correlated ( r = -0. 846, P ＜0. 01 ), as well as the diaphragm and extensor digitorum longus muscle weights ( r = - 0. 778, P ＜ 0. 01; r = - 0. 772, P ＜ 0. 01 ). (2) The levels of 3-methyl-histidine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than the COPD normal nutrition group, as well as the intervention groups B and D. The contents of tyrosine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than that of the COPD normal nutrition group,as well as the groups B and D. The body weight growth value of the intervention group B were slightly higher than the group A, without significant difference( P ＞ 0. 05 ), while the group C was significantly higher than the group A ( P ＜ 0. 01 ). Conclusions TNFα is involved in the occurrence of COPD malnutrition and skeletal muscle amyotrophy. IND can reduce the TNFα levels in the serum and the catabolic rates of the skeletal muscle proteins in malnutrition rats with COPD, so as to improve partly the skeletal muscle atrophy.

· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Membrane ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Signal Transduction ; TCF Transcription Factors ; metabolism ; Transcription Factor 7-Like 2 Protein ; Wnt Proteins ; physiology ; beta Catenin ; metabolism

Background Stromal-derived factor 1α /chemokine receptor 4(SDF-1α/CXCR4) axis is one of the important signals which mediates several different activities such as chemotaxis,adhesion,proliferation and survival resulting in recruitment to sites of immune and inflammatory reactions.Considerable evidence suggests that CXCR4/SDF-1α axis is involved in tumor angiogenesis and plays a key role in the development of ocular neovascularization.Objective The purpose of this study was to explore the effect of CXCR4 antagonist on the development of cxperimental corneal neovascularization(CNV).Methods CNV model was established in the left eye of 8-weekold clean BALB/c mouse by putting the filter with 1 mol/L NaOH at the central cornea for 40 seconds.The animals were randomizcd into hyaluronate group and CXCR4 antagonist group,and the edydrops was topically administered respectively on the day of modeling 4 times per day for 14 days.CNV was examined under the slit lamp at the fourteenth day,and then the corneal suspension and section were made.Expressions of CXCR4 mRNA and protein in corneas were detected using RT-PCR and Western blot.The CD31 level in cornea was assayed by flowcytometry and immunochemistry.The expression of VEGF in burned corneas and suspension from mouse peritoneal macrophages stimulated with CXCR4 antagonist in vitro was detected by ELISA.The use of the animal followed Ragulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Two weeks after corneal alkali burn,the growth of CNV peaked under the slit lamp.Compared with hyaluronate group,CNV was obviously decreased in the CXCR4 antagonist group.Immunochemistry showed that intensity of positive staining for CD31 in cornea in the CXCR4 antagonist group was weaker than the hyaluronate group.Flowcytometry clarified that CD31 positive cells rate was 9.50％ ±2.34％ in the CXCR4 antagonist group and 17.50％ ±3.16％ in the hyaluronate group,showing a significant difference between them (t=-7.312,P＜0.05).In 2,4,7 days after cornea alkali burn,the expressions of CXCR4 mRNA and protein were significantly enhanced in burn corneas compared with normal corneas(P＜0.01 ;P＜0.05).ELISA showed that the VEGF expression level in corneal tissue and supernatant of mouse peritoneal macrophages in vitro were significantly lower in the CXCR4 antagonist group than that of hyaluronate group(t =10.927,5.151,P＜0.05).The expression level of VEGF in corneal suspension was lower in the GM-CSH+CXCR4 antogonist group than that in the GM-CSH group (P＜0.05).Conclusions CXCR4 antagonist can reduce experimental CNV by down-regulating VEGF expression in cornea.

Objective To observe the relationship between computed tomography-gross tumor volume (GTV) and non-surgical T stage in patients with esophageal squamous cell carcinoma (ESCC) and the survival rates of patients with different GTVs,and to investigate the impact of GTV on the prognosis of ESCC after three-dimensional radiotherapy.Methods A retrospective analysis was performed on 223 ESCC patients without lymph node metastasis and distant metastasis who were hospitalized from July 2003 to January 2009.The prescribed doses of three-dimensional radiotherapy ranged from 50-70 Gy.These patients were divided into 3 or 4 groups according to different percentile intervals of GTVs.The Spearman rank correlation analysis was used for investigating the relationship between non-surgical T stage and GTV.The Kaplan-Meier method was used for calculating survival rates,and the log-rank test was used for survival difference analysis.Results The follow-up rate was 98.2％.A total of 163 patients were followed up for at least 3 years.The median GTVs of patients with T1 +2 ESCC,T3 ESCC,and T4 ESCC were 19.31 cm3,33.69 cm3,and 41.25 cm3,respectively,exhibiting a positive correlation between non-surgical T stage and GTV (P =0.000).The 5-year survival rates were 59％,43％,and 24％ (P =0.000) in 3 GTV-based groups and were 55％,51％,31％,and 24％ (P =0.004) in 4 GTV-based groups.The primary cause of death for the patients with GTVs of ≤35 cm3 and ＞35 cm3 was failure of local control (57.9％ vs 52.1％) ; 21.9％ and 13.8％ of the patients with a GTV of ＞ 35 cm3 died of uncontrol and excessive bleeding,versus 9.4％ and 3.1％ of the patients with a GTV of ≤ 35 cm3 (P =0.046 ; P =0.029) ;2 of the patients with a GTV of ＞ 35 cm3 died of grade 5 radiation pneumonitis.Conclusions GTV is positively correlated with nonsurgical T stage in esophageal cancer patients who receive radiotherapy.The 3-level and 4-level grading of GTV can be used for prognostic evaluation,and the 3-level grading of GTV is more closely related to prognosis.The patients with a larger GTV have higher incidence of uncontrol and bleeding and risk of treatment-related death than those with a smaller GTV after radiotherapy.

Objectives Todynamicallyobservethechangesofhemodynamicparametersinpatients with severe stenosis of unilateral middle cerebral artery (MCA)by transcranial Doppler ultrasound (TCD) andtoevaluateandanalyzetherelatedfactorsforinfluencingthestenoticprocess.Methods Atotalof 113 consecutive patients with severe stenosis of unilateral MCA screened by TCD and confirmed by computed tomography angiography (CTA)and digital subtraction angiography (DSA)were enrolled retrospectively. They were divided into either a progressive group (n =43 )or a non-progressive group (n=90)according to the variation of MCA hemodynamic parameters. The effects of age,sex,major risk factors for cerebrovascular disease,clinical symptoms,clinical medication,and drug compliance on the stenotic process were documented and analyzed. Results (1)The comparison of detection rate of the risk factors for cerebrovascular disease:The patients with a history of smoking (72. 1%[n=31])in the progressive group was significantly higher than that (51. 1%[n=46])in the non-progressive group (P=0.022). The period of smoking of the patients in the progressive group were longer than that in the non-progressive group (28 ± 12 years vs. 21 ± 10 years,P=0. 011). (2)Comparison of MCA hemodynamic parameters:The distal pulsatility indexes of MCA stenosis at the first diagnosis in the progressive group were all lower than those in the non-progressive group (0. 66 ± 0. 10 vs. 0. 70 ± 0. 13;t= -2. 096,P=0. 038),and the distal pulsatility indexes of MCA stenosis at the end point in the patients of the progressive group were lower than those in the non-progressive group (0. 61 ± 0. 15 vs. 0. 74 ± 0. 15). There were significant differences (t=-2. 718,P= 0. 008). The peak systolic velocity (PSV)of the progressive MCA stenotic segments at the end point in 10 patients of the progressive group was higher than that in the non-progressive group (299 ± 23 cm/s vs. 244 ± 50 cm/s,t=3. 437;P=0. 001),while PSV of MCA in 33 patients with occlusion in the progressive group were significantly lower than those in the non-progressive group (56 ± 18 cm/s vs. 244 ± 50 cm/s,t= -20. 905;P=0. 000). (3)The regular medication:The patients using statins (atorvastatin calcium)were significantly lower than those of the non-progressive group (2. 3%[n=1] vs. 54. 4%[n=49],χ2 =33. 690;P<0. 01). (4)During the follow up period,the recurrence rates of transient ischemic attack and stroke of the progressive group were significantly higher than those of the non-progressive group (27. 9%[n=12]vs. 6. 7%[n=6],32. 6%[n=14]vs. 2. 2%[n=2];all P<0.01). (5)Multivariate Logistic regression analysis showed that smokers (OR,4. 403,95%CI 1. 094-14.017),cerebrovascular event recurrence (OR,10. 648,95%CI 2. 530 -41. 261),and irregularly taking statins (OR,5. 675,95%CI 1. 631-152. 740)were all closely associated with the progress of severeMCAstenosis.Conclusion EvaluationofthehemodynamicchangesofsevereMCAstenosiswith TCD follow up study can be used as an important basis for clinical assessment of the outcomes. Stop smoking and regularly taking statins may help to delay the progress of MCA stenosis.

ObjectiveTo explore the application of diffusion-weighted magnetic resonance imaging (DWMRI) in precise radiotherapy of esophageal carcinoma.MethodsThirty-seven patients with biopsy proven esophageal cancer from March 2010 to January 2011 were included.To delineate the gross tumor volume (GTV) using CT and DWMRI images,each patient was examined by DWMRI and CT scan using the same position before radiotherapy.To compare the maximum diameters and volumes of tumor between CT and DWMRI. The tumor lengths measured by esophagogram,esophagoscope,CT and DWMRI were compared.ResultsTumor lengths under esophagogram,esophagoscope,CT and DWMRI were 5.70 cm,6.06 cm,7.97 cm and 5.79 cm respectively. The lengths between CT and esophagogram,CT and esophagoscope,CT and DWMRI had statistical significance respectively (F=4.88,P=0.003).The maximum diameters of tumor shown on CT and DWMRI were 3.79 cm and 3.81 cm respectively ( t =-0.32,P=0.751 ).The GTV were 45.75 cm3 and 38.05 cm3 in CT and DWMRI respectively (t=5.30,P =0.001 ).53 lymph nodes were assessed positive on both CT and DWMRI.DWMRI excluded 25 positive lymph nodes assesed by CT; also confirmed 15 negative lymph nodes excluded by CT,6 of which were paraesophageal lymph nodes.The addition of DWMRI information altered the clinical stage in 6 patients.ConclusionsTumor lengths measured on DWMRI and esophagogram had the optimal approximation.It was easy to find paraesophageal lymph nodes via DWMRI.With the addition of DWMRI information,the target range and clinical stage were alerted in some patients.

ObjectiveTo analyze the prognosis of 784 patients according with clinical staging of esophageal carcinoma treated with non-surgical methods,investigate the predictive value and deficiency of the clinical staging.MethodsFrom July 2003 to January 2009,784 patients with esophageal carcinoma received 3DCRT treatment.The prescribed doses ranged from 50 Gy-70 Gy with median dose of 60 Gy,1.8-2.0 Gy/fraction,1 fraction/day,5 fractions/week.65 patients received prescription dose of＜60 Gy and all the others'≥60 Gy.All the patients were divided into subgroups according to different T,N and TNM stages.Therapeutic effect was evaluated.ResultsThe follow up rate was 97.1％,503 patients were followed up for more than 3 years and 122 were followed up for more than 5 years.The 1-,3-,5-year local control rates and overall survival rates were 77.2％,54.2％,46.5％ and 69.5％,34.9％,23.9％,respectively,with median survival time of 21 months.There were significant differences of survival curves for different T stages,N stages and TNM stages.For the groups of stage Ⅰ,Ⅱ and Ⅲ,the 1-,3-,5-year survival rates were 86.4％,47.6％,45.1％ ;84.7％,46.3％,36.4％ and 64.0％,30.9％,19.1％,respectively ( x2 =29.34,P =0.000).There were 752 patients with squamous cell carcinoma ( 95.9％ )and 32 patients with non-squamous cell carcinoma (4.1％ ),the median survival time were 21 and 16 months,respectively ( x2 =4.44,P =0.035 ).There were significant difference of survival rates for the subgroups whose GTV volume ≤20 cm3,20 -40 cm3,40 -60 cm3 and ＞60 cm3 (54 months,29 months,21months and 14months,x2 =68.71,P =0.000).ConclusionsThe clinical staging of esophageal carcinoma treated with non-surgical methods could predict the prognosis accurately,for patients with different pathology and GTV volumes,there were variance in the prognosis,so we advised the complement of the two factors in the draft of clinical stages.

Objective To observe the long?term survival and adverse reactions in patients with stage T4 N (+) Ⅲ middle and lower thoracic esophageal carcinoma undergoing intensity?modulated radiotherapy ( IMRT) . Methods From 2004 to 2010, 300 patients with stage T4 N (+) Ⅲ middle and lower thoracic esophageal carcinoma, consisting of 202 treated with three?dimensional conformal radiotherapy ( 3DCRT ) and 98 treated with IMRT, were enrolled as subjects. All patients received conventionally fractionated radiotherapy with a prescribed dose of 60 Gy. The long?term survival and adverse reactions were compared between patients treated with the two different radiotherapy regimens. The survival rates were calculated by the Kaplan?Meier method and analyzed by the log?rank test. Results The 5?and 7?year sample sizes were 239 and 120, respectively. The 3DCRT group had significantly lower 1?, 3?, 5?, and 7?year local control (LC) and overall survival (OS) rates than the IMRT group (64. 4% vs. 68. 3%, 40. 6% vs. 55. 3%, 38. 3% vs. 51. 9%, 34. 2% vs. 51. 9%, P=0. 048;54. 5% vs. 63. 3%, 19. 8% vs. 34. 7%, 14. 7% vs. 24. 4%, 10. 9% vs. 20. 3%, P=0. 013) . The stratified analysis showed that for patients older than 65 years, with the length of esophageal lesion>8. 0 cm before radiotherapy, the largest diameter of esophageal lesion in computed tomography image>4. 6 cm, gross tumor volume ( GTV)>60 cm3 , metastases to adjacent tissues or organs, stage N2 , and without chemotherapy, the IMRT group had a significantly higher OS rate than the 3DCRT group (P=0. 022,0. 003,0. 022,0. 034,0. 016,0. 044,0. 047). The GTV Dmin and GTVD100 were significantly higher in the IMRT group than in the 3DCRT group ( P=0. 000,0. 000) , while the Dmax of the spinal cord was significantly lower in the IMRT group than in the 3DCRT group ( P=0. 000) . Compared with the 3DCRT group, the IMRT group had a significantly higher incidence of acute radiation?induced esophagitis, particularly grade 1?2 esophagitis (P=0. 000). The mortality rate caused by local tumor was significantly higher in the 3DCRT group than in the IMRT group ( P= 0. 039 ) . Conclusions In the treatment of locally advanced middle and lower thoracic esophageal carcinoma, IMRT is safe and effective;it significantly improves the LC rate and long?term survival without severe toxicity to normal tissues. The results of this retrospective study need to be confirmed by prospective randomized controlled studies.