Objective To investigate the regulation effect of promoter methylation of deathassociated protein kinase (DAPK) on mRNA and protein expression of DAPK in tissue of primary gastric cancer (GC). Methods The cancerous and noncancerous samples from 62 patients with GC were determined by RT-PCR for mRNA expression of DAPK. The DAPK promoter methylation was detected by methylation-specific PCR. The protein expression of DAPK in 34 patients with methylation was determined by Western blot. Results mRNA and protein expre.ssions of DAPK in cancerous tissues were reduced significantly compared to noncancerous tissues (0. 2863d±0. 2027 vs 0. 57364±0. 1968,0. 2616±0. 0913 vs 0. 65294±0. 1808, P＜0.01). Methylation frequency of DAPK in cancerous tissues was higher than that in noncancerous tissues (54.8% vs 17.7%, P＜0.01). Furthermore, DAPK mRNA expression was decreased in methylation group compared to unmethylation group (0.1399±0. 0835 vs 0. 46404±0. 1569, P＜0. 01). Moreover, a significant correlation was demonstrated between the TNM stage and DAPK promoter methylation (P = 0. 04). Conclusion Expression of DAPK is down-regulated in cancerous tissues at mRNA and protein levels. Low expression of DAPK is associated with hypermethylation of the promoter of DAPK gene.

Objective To evaluate clinical value of utilization of real‐time fluorescence isothermal RNA amplification assay (SAT)for detecting Mycobacterium tuberculosis(MTB) in pleural effusion samples of patients with tuberculous pleurisy .Methods 90 hydrothorax specimens of patients with tuberculous pleurisy and 52 hydrothorax specimens of patients with non‐tuberculous pleurisy were selected ,and MTB was detected by using Roche culturing method ,SAT and TB DNA fluorescence quantitative expan‐sion method respectively .The sensitivity ,specificity and clinical value of SAT for detecting MTB were analysed .Results Taking Roche culturing method as the standard ,the sensitivity ,specificity and Youden index of SAT for detecting MTB were 90 .9% , 72 .1% and 0 .630 respectively .In patients clinically diagnosed with tuberculous pleurisy ,the positive detection rate of hydrothorax specimens detected by using Roche culturing method and SAT were 24 .4% and 43 .3% respectively ,and had statistically significant differences(χ2 =7 .166 3 ,P=0 .001 4) .Conclusion SAT is a rapid ,sensitive and specific method for the detection of MTB in pleu‐ral effusion samples ,which could improve the detection rate of MTB and decrease incidence of missed diagnosis and misdiagnosis .

Objective To detect the levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-l (sICAM-1) in the sera of patients with systemic lupus erythe-matosus (SLE) and their clinical significance was analysed. Methods Serum level of sVCAM-1 and sICAM-1 of 30 controls and 60 SLE patients were measured by enzyme linked immunosorbent assay (ELISA). Results 1 Serum levels of sVCAM-1 were significantly increasd in patients with SLE compared with those in normal controls (P

Objective To investigate the relationship between the serum level of soluble vascular cell adhesion molecule-1(sVCAM-1) and soluble intercellular adhesion molecule-1(sICAM-1) and the disease activity in systemic lupus erythematosus(SLE). Methods The serum concentrations of sVCAM-1 and sICAM-1 were measured by ELISA in 60 SLE patients and age- and sex-matched normal controls. Results ① Serum levels of sVCAM-1 and sICAM-1 were significantly increased in SLE patients compared with those in normal controls (P

Objective:As the function of ??T cells and NK in the immunological therapy is showing more and more important,the characteristics of ?? T cells,NK and LAK were analyzed comparatively after that they were richened by isolating and incubating in vitro.Methods:The cells were collected using MACS after the cells were panned respectively with special monoclonal antibodies.Then the characteristics including proliferation,phenotype,cytotoxin and the down regulation blocked by specific antibodies were analyzed.Results:The ?? T cells can expand 600～800 times after culturing 2 weeks and the percent of CD3,CD8 and ?? expressed on the collected cells were 72.29%,58.02% and 65.98% respectively ?? T cells showed the high cytotoxin to K562(NK sensitive cell line),Raji and XG 7 cell lines(NK non sensitive cell lines).The percentage of cytotoxin reached 35.98%,52.27% and 69.08% respectively compared with ??T respectively.No obviously change of percent of ??T cytotxic ability to these target cells were observed using special MHC class I monoclonal antibody to block the ??T cell before coculturing the target cells with ?? T cells.Conclusion:All of ?? T cells,NK and LAK showed the non specific cytotoxin to tumor cells.The cytotoxic capability of ?? T cells did not be effected after blocking with MHC class I monoclonal antibody.These results demonstrated that ?? T cell could kill more kinds of tumor cells than NK and LAK.

Objective To investigate the motion characteristics of primary thoracic esophageal carcinoma with four-dimensional computed tomography(4DCT).Methods Sixteen patients with primary thoracic esophageal carcinoma received respiratory gated 4DCT imaging,mapping the GTV1-GTV10 on every patient's each subsequent CT image of 10 images in the full-respiratory phase,and measuring the displacement of each centre of GTV.These displacements and directions were analyzed on different segments of esophagus.Results The mean total lung volume and GTV volume was 2993.5 cm3,35.00 cm3 and 3362.12 cm3,34.84 cm'respectively on end-expiration and end-inspiration phases(t=12.36,P=0.000and t=-0.61,P=0.546).The total mean peak to peak displacement of GTV were 0.65 mm,0.55 mm,and 2.03 nnn in x,y-and z-axis direction,respectively(F=41.14,P=0.000).The motion in x-axis,y-axis and z-axis were 0.50 mm,0.48mm,1.23 mm in the upper segment(F=5.45,P=0.017),0.68 mm,0.62 mm,1.97 mm in the middle segment(F=27.74,P=0.000),0.72 mm,0.38 mm,3.05 mm in the lower segment,respectively(F=15.61,P=0.000).Conclusions The displacement of tumor in z axis is more notable than x-,y-axis in thoracic esophageal carcinoma.The displacement of tumor x-,y-and z-axis is different in different segment of thoracic esophageal carcinoma.

Objective To optimize the registration template of kilovohage cone-beam CT (CBCT) guided radiotherapy in whole breast irradiation(WBI)after lumpectomy of breast cancer.Methods From April 2006 to July 2009,twelve patients undergoing WBI with intensity-modulated radiotherapy (IMRT)were recruited in this study.All patients were performed with both conventional planning CT and CBCT integrated on Varian 23 EX.Six distinguishable referenee points(the diameter 1 mm)around the lumpectomy cavity and the surrounding gland on the planning CT image were marked.The images were manually registered offline based on the breast surface,surgical clips,breast gland,contiguous rib,ipsilaterai lung and its external contours,respectively.The same six reference points were then marked on the CBCT image.The performance of the five registration templates was compared using the concept of registration error,while the registration time was taken into account.The registration error was calculated based on the six reference points'translations between the planning CT image and CBCT image,and analyzed with SPSS 13.0 software using one-way ANOVA.Results The values of the registration error for the breast surface,surgical clips,breast gland, contiguous rib,ipsilateral lung and its external contours were(0.60±0.20),(0.43±0.15),(0.49±0.19),(0.69±0.36)and(0.94±0.49)cm,respectively,and the registration time were(3.8±1.1),(3.0±0.9),(4.7±1.7),(4.3±1.3)and(4.5±1.3)min,respectively.There was no statistical difforence between the breast surface,surgical clips and breast gland registration template(t=0.48-1.36,P>0.05),the same result trend to contiguous rib compared with ipsilateral lung(t=2.00,P=0.055),however,there was significant difference between surgical clips and the last two registration methods(t=2.08-4.08,P<0.05).Conclusion In this initial study with a modest number of patients,surgical clips show a best registration template from the standpoint of accuracy and efficiency,whereas contiguous rib and ipsilateral lung are not an ideal method.

Objective To study the effects of postnatal overfeeding and high-fat diet on blood pressure of rats,and to explore the pathophysiological mechanism underlying hypertension induced by continuous early postnatal overfeeding.Methods Male Sprague-Dawley rats were randomly divided into normal feeding group (10/litter) and overfeeding group (3/litter) on postnatal day 3 with a random number table.After weaning at postnatal week 3,the rats were randomly given standard chow or high-fat (HF) diet until week 16.Hence four groups were analyzed,namely normal feeding group,breastfed overfeeding group,post-weaning overfeeding group,and continuous overfeeding group.Body weight was continuously monitored in each week.Visceral fat pad (retroperitoneal and perigenital),systolic pressure,and heart rate were observed at week 3 and week 16.Thoracic aorta was sampled for measurement of vascular endothelial dilation function.Histological morphology was observed with HE staining,nitric oxide content of thoracic aorta was detected with nitrate reductase method.The mRNA expression of endothelial nitric oxide synthase (eNOS) in thoracic aorta was determined by real-time polymerase chain reaction.The protein expressions of eNOS and phosphorylated eNOS were determined by Western blot.Results At week 3,breastfed overfeeding rats displayed significantly larger body weight [(77.80 ± 0.57) g vs.(62.80 ±0.85) g,t =14.576,P ＜ 0.01] and visceral fat [retroperitoneal:(8.19 ± 0.49) mg/g vs.(4.92 ± 0.31) mg/g,t =5.629,P＜0.01;perigenital:(3.50 ±0.29) mg/g vs.(2.08 ±0.13) mg/g,t =4.552,P ＜0.01] compared with normal feedindg rats,and the protein expression of phosphorylated eNOS in aortic tissues was significantly reduced to week 16 (F =15.215,P ＜0.01);high-fat diet feeding after weaning further increased the body weight and fat mass in breastfed overfeeding rats.At week 16,continuous overfeeding rats showed hypertension [(149 ± 1.94) mmHg (1 mmHg =0.133 kPa),F =22.834,P ＜0.01],impaired vascular endothelial dilation function (F =7.648,P ＜ 0.05),and reduced protein expression of phosphorylated eNOS (F =15.215,P ＜ 0.01),while the post-weaning overfeeding group only had elevated blood pressure.Conclusions Overfeeding in breastfeeding period and high-fat diet after weaning leads to hypertension.The continuous decrease in phosphorylated eNOS in vascular tissues may be an important molecular process participating in the occurrence of vascular endothelial dysfunction in adults induced by postnatal overfeeding.

Objective To evaluate the effects of repetitive noxious stimuli during the neonatal period on the pain sensitivity during adulthood in rats.Methods Twenty pathogen-free male neonatal SpragueDawley rats,weighing 6.2-6.8 g,were randomly divided into 2 groups (n=10 each) using a random number table:control group (group C) and repetitive noxious stimuli group (group RNS).In RNS group,neonatal rats sequentially received needle pricks into the dorsal surface of 4 paws at 6 h intervals with a 28 G needle per day during the first postnatal week.In group C,the animals received non-painful tactile stimuli with cotton tip rub.At the age of 10 weeks,complete Freund' s adjuvant was injected into the plantar surface of the right hind paw to induce inflammatory pain.Before injection,and at 24 h and 7 days after injection (T0-2),the mechanical and thermal thresholds were measured,and alcohol preference test was performed for assessment of the behavior at T2.Results The mechanical threshold of the right paws was significantly lower at T1.2 than that of the left paws in the two groups.Compared to group C,the mechanical threshold of the left and right paws at T0-2 and thermal threshold of the right paws at T1 were significantly decreased,and alcohol intake was increased in group RNS.Conclusion Repetitive noxious stimuli during the neonatal period can lead to increase in the pain sensitivity during adulthood in rats.

0.05). Conclusions TVT is a safe, effective and minimally invasive surgical procedure for treatment of female stress urinary incontinence.