Objective:To investigate the diagnostic value of telomerase activity and p16 gene methylation from exfoliated cells of sputum in 55 cases of solitary nodules and of being suspected of early peripheral lung cancer(T1N0M0).Methods:The sputum specimens from 34 cases of cancer nodes and 21 cases of benign lung lesion were detected for telomerase activity and p16 gene methylation by methylation analysis.Results:The qualitative diagnosis accuracy of CT scan was 61.8%(34/55) for peripheral lung cancer.Telomerase activity was positive in 29 cases:Sensitivity was 79.4%;Specificity was 90.5%;and accuracy was 83.6%.p16 gene methylation was in 11 cases:Sensitivity was 32.4%;specificity was 100%,and veracity was 58.2%.The sensitivity was increased to 86.1% by the combination of telomerase activity and p16 gene methylatiion.Conclusion:The results suggest that combining CT scan with telomerase activity and p16 gene methylation detection in sputum for patients with lung cancer may enhance the diagnostic value of conventional cytology.It is a promising approach to early diagnosis of lung cancer and massive screening in terms of its rapidity,economy and simplicity.

Objective To evaluate the efficacy of a self-expandable metal stent coated with autologous endothelial progenitor cells(EPCs)for prevention of restenosis in transjugular intrahepatic portosystemic shunt(TIPS)in a swiue model.Methods EPCs were coated on the metal stents using fibrin gel before TIPS procedure.TIPS was performed in 15 young adult pigs,using an autologous EPC-seeded stent(treatment group,n=9)or a conventional bare metal stent(control group,n=6).All pigs were sacrificed at 2 weeks after TIPS procedure.Portography was performed immediately before the euthanasia.Gross and microscopic pathological exams and immunohistochemical exams of the TIPS track specimens were performed.Fisher test and t test were used to analyse the data.Results TIPS was performed successfully in all the 15 swine.On day 14 of follow-up,direct portography and necropsy demonstrated that 5 shunts remained patent,2 shunts stenosed,and the remaining 2 shunts occluded in the treatment group(n=9);while 5 shunts were occluded and one shunt was stenotic in the control group(n=6).The patency rate was 56%vs 0(P=0.03)between the two groups.Histological analyses showed a greater pseudo-intimal hyperplasia in the TIPS track of the control group than that of the treatment group(pseudointimal thickness at hepatic vein,hepatic parenchyma and portal vein site was(1.2±0.4),(1.3±0.5),(1.5±0.4)mmvs(1.0±0.6),(0.9±0.5),(1.0±0.4)mm respectively(P＜0.05).Conclusion The EPC-coated metal stent is feasibly constructed in vitro and improves the patency in TIPS in a porcine model.

BACKGROUND:Superparamagnetic iron oxide (SPIO) labeling can trace the migration of stem cells in vivo, and the fluorescent DiI dye is suitable for marking and tracing cells because of its less influence on cellviability, proliferation and differentiation.
OBJECTIVE:To observe the effect and safety of SPIO and fluorescent DiI dye to double label bone marrow mesenchymal stem cells.
METHODS:The bilateral lower limbs of rats were isolated sterilely. Bone marrow was obtained by rinsing using low-glucose DMEM. Bone marrow mesenchymal stem cells were isolated by the whole bone marrow adherence method and purified by differential attachment method. Purified cells were dual-labeled with SPIO particle and fluorescent DiI dye.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells could be separated at 8-10 days after primary culture and the subculturing cycle was 3-4 days. Rat bone marrow mesenchymal stem cells could be effectively labeled with SPIO-DiI and the labeling efficiency was almost 100%. Blue irons contained in intracytoplasmatic vesicles could be observed clearly with Prussian blue staining, and the fluorescence microscopy showed red fluorescence at cytoplasm. Survival and apoptosis percentages obtained by MTT analysis were similar among labeled and unlabeled bone marrow mesenchymal stem cells that were both about 95%.These findings indicate that the rat bone marrow mesenchymal stem cells could be efficiently labeled with SPIO-DiI to construct a nano-bioprobe, without significant changes in morphology, viability and proliferation.

Objective The main purpose of the present study was to reveal the correlation among the CT and p53?p16 protein of non-small cell lung carcinoma(NSCLC).Methods The expression of p53?p16 protein in 52 cases by SABC immunohistochemical technique and the relationship to CT appearance were analysed.Results (1)The experssion of p53 protein of NSCLC was significantly higher than that of their adjacent tissues and their normal lung tissues(?

Objective To evaluate biochemical characteristics and the trend of diabetic complications in patients with newly-diagnosed type 2 diabetes from 1994 to 2008. Methods We utilized the database of the diabetes complications assessment and analyzed the metabolic disorder and the diabetic complications in the patients with newly diagnosed diabetes. Results 2 085 cases were collected, including 1189 males and 896 females. The average age of onset of diabetes was 51.6±13.1 and 54.6±7.9 yrs respectively in 2008 and 1994. During 1994,no case was found in subjects aged 20-29 yrs and 5% of the patients were aged 30-39; but 2% of patients aged 20-29 and 16% aged 30-39 yrs were found in 2008. BMI was increased from 24.48±4.15 in 1994 to 26.03±3.63 in 2008. Percentage of patients with abnormal BMI ( ≥25 kg/m2 ), WHR [≥0.90 (male) or ≥0.85 (female)]increased significantly from 63.6%, 75.0%, and 71.4% in 1994 to 79.6%, 95.2%, and 93.8% in 2008,respectively. Both SBP and DBP were not significantly changed. The fasting blood and postprandial blood glucose,HbA1c decreased from 10.3 mmol/L, 15.2 mmol/L, 11.1% in 1994 to 9.0 mmol/L, 14.3 mmol/L, and 8.6% in 2008, respectively. The average TG level increased from 1.7 mmol/L in 1994 to 2. 1 mmol/L in 2008,however, TC and HDL level were not significantly changed. The prevalence of diabetic retinopathy decreased from 28.2% in 1994 to 3.9% in 2008. The prevalence of diabetic nephropathy increased from 17.7% in 1994 to 24.6% in 2008. The prevalence of diabetic cardiovascular disease increased from 14.3% in 1994 to 24. 1% in 2008. Compared with the patients without microvascular complications, the patients with microvascular complications had higher SBP, DBP, and HbA1c( 136/78 vs 130/77 mm Hg, 9.41% vs 9.11% ). The patients with macrovascular complications had older age, higher SBP, TC, and TG than those without macrovascular complications (53.4 vs 50.0 yrs; 132 vs 129 mm Hg ; 5.3 vs 5.1 mmol/L and 2.6 vs 2.1 mmol/L). Conclusions In the studied newly-diagnosed diabetic patients from 1994 to 2008, there were increasing incidences of obesity and hypertriglyceridemia. However, the prevalence of diabetic retinopathy decreased significantly, while that of nephropathy showed no significant change.Cardiovascular complications were markedly increased.

Objective:To explore the changes of number of CD4+ CD25 + foxp3 + regulatory T cells (Treg)and natural killer cells (NK)in the peripheral immune organs and tumor tissue of the murine models of lung cancer,and to clarify their effects on the development of lung cancer.Methods:The C57BL/6 mice were divided into Lewis lung carcinoma cells (LLC)injection group and normal control group.The mice in LLC injection group were injected with LLC subcutaneously in the armpit to establish the tumor models,while the mice in normal control group were injected with the same amount of saline.The number of CD4+ CD25 + T cells,CD4+ CD25 + foxp3 + Tregs in the spleen,lymph nodes and lung cancer tissues,and the number of NK cells in the spleen tissue were labeled by cell surface or intracellular antibody staining,and detected by flow cytometry.Results:The ratios of CD4+ CD25 + T cells to CD4+ T cells,foxp3+ cells to CD4+ CD25 + T cells,and the number of CD4+ CD25 + foxp3 + Treg in the spleen and lymph nodes of the mice in LLC injection group were increased significantly compared with normal control group (P <0.05 or P <0.01).Moreover,the ratios of CD4+ CD25 + T cells to CD4+ T cells and foxp3 + cells to CD4+ CD25 + T cells in the tumor tissue were significantly higher than those in the spleen and lymph nodes of the mice in LLC injection group.However,the ratio of NK cells in the spleen tissue of the mice in lung cancer group was significantly decreased compared with normal control group (P <0.05).Conclusion:The increase of ratio and the number of Treg cells and the decrease of ratio of NK cells in the main immune organs of lung cancer mice may promote the development of tumor and inhibit the immune response to cancer cells in vivo .

Objective To observe the effects of grape seed proanthocyanidin extract (GSPE)combined with atorvastatin on carotid atherosclerosis of the elderly. Methods Consecutivly 122 patient aged( 73.2 ±12.8) years with hypercholesterolemia and diagnosed as asymptomatic carotid artery plaques were randomly assigned to single atorvastatin(20 mg/d) treatment group (n=63) and combination treatment (atorvastatin 10 mg/d plus GSPE 400 mg/d) group (n=59).Serum total cholesterol (TC),triglyceride (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C) and high-sensitive C-reactive protein (hs-CRP) levels were detected before treatment and 3,6,12 months after treatment.Mean maximum carotid intima-media thickness (CIMT),plaque score and stability were also assessed by carotid ultrasound. Results After 3 months of treatment,serum levels of TC,TG and LDL-C in both groups were lower than before treatment (all P＜0.01),but no difference was found between the two groups (P＞0.05).HDL-C levels was enhanced by 20.2％ in combination treatment group after 3 months of treatment (P＜0.05),there was difference after 6 months treatment between two groups (P＜0.05).Serum hs-CRP concentrations were decreased by 1.7 mg/L and 3.6 mg/L,3.8 mg/L and 6.3 mg/L,5.9 mg/L and 8.3 mg/L after 3,6 and 12 months treatment as compared with pretreatment,respectively in the two groups (P＜0.05 or P＜0.01).There was difference in hs-CRP levels between groups after 6 and 1.2months of treatment (P＜0.05 or P＜0.01).There were MMCIMT reduction by 1.3％ and 2.0％ (P＞0.05),3.4％ and 5.3％(P＞0.05 and P＜0.05 ),5.1％ and 8.6％ after 3,6 and 12 months of treatment(P＜0.05 and P＜0.01) in the two groups,with distinct difference after 12 month treatment (P＜ 0.01 ).The plaque scores were declined by 6.8 ％ and 13.1％ ( both P ＞ 0.05 ),14.5 ％ and 28.0％ (P＞0.05 and P＜0.05),19.2％ and 45.0％(P＜0.05 and P＜0.01)after 3,6 and 12 months of treatment while significant difference was found between the two groups after 12 months treatment (P＜0.05).Unstable plaque and the total number of plaque was dropped by 8 and 14 in single group,19 and 28 in combination group,respectively,after 12 months of treatment. Conclusions GSPE combined with atorvastatin may inhibit the development of carotid intima-media thickness,reduce carotid plaque and promote stabilization of carotid plaque.

According to the alanine aminotransferase (ALT)level, 4 509 patients were assigned into group A (n=449, with raised ALT)and group B (n=4 060, normal ALT). Between the patients of group A and B, differences existed in age [(48.5 ± 11.3 vs 55.7 ± 11.4) years, P＜0.01], duration of diabetes [( 36.8 ±45.0 vs 56.2±58.8 ) months, P＜0. 01], body mass index[BMI, (27.7±3.9 vs 25.8±3.4) kg/m2, P＜0.01], waist-tohip ratio (0.95±0.06 vs 0. 93±0.07, P＜0. 01 ), diastolic blood pressure [( 78± 10 vs 75± 10) mm Hg, 1 mm Hg=0. 133 kPa, P＜0. 01], fasting blood glucose [(9.04±2.91 vs 8.63 ±3.05 ) mmol/L, P = 0. 008], 2 h blood glucose after meal[( 13.85±4.67 vs 13.07 ± 4. 92 ) mmol/L, P=0. 002], HbA1c(8. 11% ± 1.82% vs 7.74% ±1.96%, P＜0. 01 ), fasting serum insulin[( 10.59±7.31 vs 7.97±7.18) mU/L, P＜0. 01], postprandial insulin [(48.96±43.80 vs 35.25 ±32.37 ) mU/L, P＜0. 01], homeostasis model assessment for insulin resistance ( HOMA-IR, 4.11±-2.85 vs 3.00 ± 2.92, P＜ 0. 01 )、 triglycerides [( 2.77 ± 2.50 vs 2. 19 ± 2.99 ) mmol/L, P＜0. 01], and high-density lipoprotein-cholesterol [HDL-C, ( 1.20 ± 0. 30 vs 1.29 ± 0. 83 ) mmol/L, P = 0. 01].Logistic regression analysis showed that HbA1C, postprandial insulin, and HOMA-IR, uric acid and urinary albumin were positively, and HDL-C negatively related with the ALT level. It suggests that raised ALT seems to be an index related to the clustering of cardiovascular risk factors, insulin resistance, and earlier onset of type 2 diabetes.

BACKGROUNDTo investigate the diagnostic value of chest CT scan combined with telomerase activity and p16 gene methylation from exfoliated cells of sputum in 55 cases of solitary pulmonary nodules (SPN; ≤30 mm)suspected early peripheral lung cancer.

METHODSThe sputum specimens from 34 cases of cancer nodules and 21 cases of benign lesion were detected for telomerase activity by TRAP-PCR-ELISA and p16 gene methylation by PCR-based methylation analysis.

RESULTSThe qualitative diagnostic accuracy of CT scan was 61.8%(34/55) for SPN provided by pathology. Cytology analysis of sputum was positive in 13 cases (38.2%). Telomerase activity was positive in 29 cases: sensitivity was 79.4%, specificity was 90.5%, accuracy was 83.6%; p16 gene methylation was found in 11 cases: sensitivity was 32.4%, specificity was 100.0%, and accuracy was 58.2%. The sensitivity was increased to 86.1% by combination of telomerase activity and p16 gene methylation. Compared with nodules without malignant CT signs, expression of telomerase activity and p16 methylation of SPN with malignant CT signs (lobulation or spiculate protuberance or spicule sign) had a significant difference (P < 0.01).

CONCLUSIONSThe results suggest that chest CT scan combined with telomerase activity and p16 gene methylation detection in sputum for patients with peripheral lung cancer may enhance the diagnostic value of radiology and conventional cytology.