Objective To investigate the effect of a disintegrin and metalloprotease domain(ADAM8)gene on colon cancer HCT8 cell proliferation and proliferation signal transduction pathways PI3K-Akt-mTOR. Methods Colon cancer HCT8 cells were cultured in vitro,and transfected with ADAM8 overexpression plasmid and RNA interfering plasmid. Cell proliferation were detected by EdU and MTS method assays. PI3K activity was measured by PI3K activity detection kit,Western Blot method was performed to detect the ratio of Akt and p-Akt and expression of mTOR. Results Compared with control group(1.00 ±0.12),the cell proliferation in ADAM8 overexpression group(1.22 ±0.13)was significantly higher (P<0.05)and in RNA interfering group(0.78 ±0.11)was significantly lower while PI3K activity had no significant changes in three groups. After ADAM8 overexpression,and the ratio of p-Akt/Akt and mTOR expression were increased significantly,while reduced significantly after RNA interferered. Conclusion ADAM8 can promote HCT8 cell proliferation through enhancing the phosphorylation of Akt and promoting the expression of mTOR.

Objective To explore the effect of mouse proliferation-associated protein 2G4 (p38-2G4) high-expression on the proliferation and erythriod differentiation of murine erythroleukemia ( MEL ) cells.Methods To establish the recombinant lentivirus vector p 38-2G4-pLJM1, the p38-2G4-pLJM1 was cotransfected into HEK293T cells to obtain lentivirus with pCMV-VSV-G and pCMV-dR8.2.Lentivirus were infected into MEL cells to establish the stably p 38-2G4 high-expressed MEL cells.Western blotting was used to analyse the high-expression efficiency.MTT assay and benzidine staining were applied to detect the cell viability and hemoglobin synthesis of the stable cell line in presence /absence of inducers.Results Western blotting showed that the p38-2G4 high-expression stable cell strain had a higher expression of p38-2G4 as compared to the control group ( MEL) ( P <0.05).MTT result showed that there was no difference between the p38-2G4 high-expression cell strain and the control group (P>0.05), but the hemoglobin synthesis had been reduced as compared to the control group (P<0.05).Conclusion p38-2G4 high-expression does not affect the cell viability of MEL cells , but inhibits the erythriod differentiation of MEL cells in three independent experiments .

Objective To evaluate the impact of 1125 seeds para-tracheal braehytherapy on regional tissue injury in rab-bit models. Methods 42 rabbits were randomized into 7 groups. Group 1 to 6 belong to study groups (in which 1,4,5 and 6 belong to "dose gradient" subgroup, while 2,3 and 4 to "chronologic" subgroup) , while the last group acts as negative con-trol. The activity of seeds in study group were 0.3 mCi in group 1, 0.5 mCi in group 2 to 5, 0.7 mCi in group 5, and 0.9mCi in group 6. False seeds (0 mCi) were used for the negative control. 4 seeds with equal dosage were implanted between trachea and esophagus in each rabbit under general anesthesia. Seeds arrangement was made according to Paris principle. For the tissue injury evaluation, group 2 was sacrificed by the end of first month post-operatively, group 3 at the end of the second month, and group 4 end of the third month. The rest of rabbits were also sacrificed at the end of the third month. Pieces of adjacent e-sophagus and trachea were sampled from each rabbit. Tissue injury features such as inflammation, edema, congestion or fibrosis as evaluated histologically. Results All rabbits were healthy during study period except 5. Histological analysis revealed that trachea samples from all groups had lymphocytas and plasma cells infiltration as signs of chronic inflammation, hut fibrosis was nut clearly visible. There were no differences between study and control groups with respect to inflammation, edema and con-gestion scores. But in groups which received the highest doses of radiation or sacrificed at 60 d showed more eosinophil infiltra-tion and epithelum degeneration, and statistical significance was reached between these groups and control. Esophageal samples had less histological changes compared with trachea. Conclusion Para-tracheal implantation of ~(125)Ⅰ seeds with therapeutic or higher dosage only induce minor and reversible damage to the regional tissue. This implies that ~(125)Ⅰ implants adjacent to trachea or esophagus are clinically safe.

ObjectiveTo evaluate thd cell biologic changes in thd non-small-cell lung cancer(NSCLC) which PTEN gene were activated by double-stranded RNA(dsRNA).MethodsSpecific dsRNA was designed.First,the promoter region of PTEN gene was determined by Promoter 2.0 program,then the CpG island in the promoter was found by CpGisland searcher software and the possible target non-CpG sequence that dsRNA might activate were defined by SiRNA Target Finder software.dsRNA were synthesized at Genechem Company( Shanghai,China).Then the specific dsRNA was transfected into A549 and H292 cells which were stored in our laboratory using Lipofectamine 2000 ( Invitrogen,USA) according to manufacture's instruction.Total celluar RNA was isolated.The expression of PTEN mRNA in transfected,control and mock group were determined by real-time quantitative polymerase chain reaction.Cell profiferation was investigated on days 1 to 5 by using Cell Counting Kit-8 according to the manufature's technical manual.Cell invasion ability was assessed by Transwell method that transmembrane cells were counted,and cell bycle distribution were studied by flow cytometer(FCM) using CycleTESTTM PLUS DNA Reagent Kit.ResultsAfter the introduction of dsRNA into the A549 cells,the PTEN mRNA expressin was upregulated to (4.35 ±0.42) folds compared with the mock and control cells.And in H292 cells,the mRNA expression of PTEN was upregulated to (3.92 ± 0.20) folds.It confirmed the RNA activation phenomenon in the PTEN gene in NSCLC cells.Compared with the control group,the number of alive transfected cells did not decreased in the cell proliferation assay.In the cell invasion test we found that the transmembrane A549 cells were 122.4 ±11.2 vs.150.7 ±13.1 in transfected group and control group respectively.In the cell cycle distribution we found dsRNA in duced part ofthe transfected cells arrested in G1 phase and a corresponding decrease in S-phase population was observed,though this change was not statistically significant.Conclusion The expression of PTEN mRNA could by enhanced by inducing the specific dsRNA into the A549 and H292 cells,though no evidence was found that after the activation of silenced PTEN,the cell proliferation and invasion ability were significantly changed.

Objective To improve our understanding of localized Castleman's disease ( Localized Castleman's disease, LCD) ,and to improve its diagnosis and treatment. Methods Clinical characteristics and treatment of 26 LCD cases were retrospectively analyzed, and its clinical features and treatment strategies were reviewed. Results Among the 26 cases, there were 10 cases with clinical symptoms, which mainly showed local pain induced by the compression of the tumors, and 3 in the 10 cases associated with paraneoplastic pemphigus. The swollen lymph node was at a localized area, which was mostly at retroperitoneal (10 cases) and mediastinum (7 cases). The CT scan of LCD had its special characters including local calcification and necrosis. 22 cases were of hyaline vascular type, and the other 4 cases were of plasma type based on histopathologic examination. Twenty-five patients received complete tumor resection and 2 cases of them recurred after a follow-up of 5 to 206 months averaging at 48 ± 13 months. In one case the tumor adjoining vital organs deep in the mediastinum couldn't be completely resected. This patient and another with complete tumor resection recurred and received combined chemotherapy with complete tumor disappearance and were all alive without recurrence as found by follow up to May, 2010. The other patient with recurrent tumor after tumor resection didn't receive chemotherapy and died 11 years later. Conclusions LCD patients mainly have isolated lymphadenectasis, and some patients may have systemic symptom and show abnormal laboratory results. CT scan is helpful in establishing a diagnosis of LCD.Complete surgical resection offers a favorite result for this disease.

Clinical features and related information on diagnosis and treatment of 45 cases of Castleman's disease (CD) were retrospectively analyzed.Based on the clinical classification, localized CD (LCD) was found in 26 cases, multicentric CD (MCD) was found in 19 cases.Most cases of LCD presented the symptoms of compression, while MCD had complicated and non-specific clinical manifestations, making the early diagnosis more difficult.All 26 cases with LCD underwent surgery, among which only 2 cases relapsed.Sixteen out of 19 patients with MCD were treated with glucocorticoids or combined chemotherapy, and 14 cases achieved complete or partial remission.The results show that patients with CD have variant manifestation and the diagnosis depend on CT scan or histopathology examination.Most LCD can be cured by complete surgical resection, and MCD can achieve remission by the treatment with glucocorticoids or combined chemotherapy.

Objective To explore disease-associated proteins in the serum of patients with inflammatory bowel disease by serum proteomic analysis combined with mixed sampling strategy. Methods The serum proteins from 8 healthy adults and 8 patients with inflammatory bowel disease who had been admitted to the Sixth Affiliated Hospital of Sun Yat-sen University from March 2007 to June 2008 were collected. Two-dimensional differential in-gel electrophoresis (2D-DIGE) was used to define patterns of protein expression after the serum proteins were cross-labeled with cariant CyDye. Proteins that showed differential expressions were analyzed by matrix-assisted laser dcsorption/ionization time of flight mass spectrometry. The 2D-DIGE images were analyzed using DeCyder V6.0 software, and the differences between the groups were analyzed by t-test. Results Maps of 2D-DIGE of patients with inflammatory bowel disease and healthy adults were obtained. Fifty-six spots of proteins with abnormal expression were detected in patients with inflammatory bowel disease, and 30 proteins were identified using mass spectrometry and database retrieval. The 30 proteins included haptoglobin, complement factor B, apelipoprotein A- Ⅱ precursor and GTPase K-ras. Conclusions Serum proteomic analysis combined with mixed sampling strategy can clearly detect the difference in the expression of serum proteins between patients with inflammatory bowel disease and healthy adults. The differentially expressed proteins may provide new biornarkers for investigating the biological behavior of inflammatory bowel disease.

Objective To explore the differentially expressed proteins during erythroid differentiation .Methods The murine erythroleukemia ( MEL) cell were treated by DMSO , and the comparative proteomic was systematically analyzed and identified on different differentiating time points .ratio of cell differentiation and viability were detected by benzidine staining, MTT assay and Ter119 immunofluorescence.Using two-dimensional gel electrophoresis combined with mass spectrometry technology and bioinformatics analysis , we conducted a comparative proteomic analysis on MEL cells during the process of induced differentiation to screen and identify differential proteins .Results The MEL cells induced by 1.2%DMSO for 0 hour, 6hours, 12hours, 24hours, 36hours, 48hours, 72 hours, 96 hours, 120 hours were collected for proteomic analysis, by two-dimensional gel electrophoresis combined with mass spectrometry .A total of 87 kinds of proteins were successfully identified .MEL cells exposed to DMSO at a final concentration of 1.2% for 120 hours reached the highest differentiation rate of 67%.MTT assay showed that 1.0%, 1.2%, 1.4% DMSO had no inhibiting effect on cell vitality.Conclusion DMSO may induce MEL cells to differentiate and have no inhibiting effect on cell vitality .The 87 kinds of differentially expressed proteins from two-dimentional gel electrophoresis may be divided into twelve categories ;the most three parts are 41%enzyme protein, 15%structural protein and 13%regulatory protein.

Objective To evaluate the feasibility of lobectomy by completely Video-Assisted Thoracoscopic Surgery (cVATS) in the management of bronchiectasis.Methods Between June 2001 and October 2010,a total of 60 major lobectomies were performed in our single center on 32 female and 28 male patients of bronchiectasis,with a mean age of 43.4( range 17 to 69)years.All lobectomies were carried out anatomically and divided into thoracotomy group and cVATS group.Pulmonary vessels and bronchus were dissected by endo-cutters.Conversion to a thoracotomy took place if severe adhesion or bleeding was encountered.Results The operations included 5 lobectomies of right upper lobe,3 of middle lobe,6 of right lowerlobe,3 of left upper lobe,26 of left lower lobe,10 of left lower lobe plus lingular segment,4 of left pneumonectomy,1 of bi-lobectomy,1 of right middle lobe plus wedge resection of lower lobe and 1 of left lower lobe plus right middle lobe.There were 25 patients in the thoracotomy group and 35 patients in the cVATS group,in which 2 operations (5.7％) converted due to severe adhesion,poor differentiation of the fissure and/or the proliferation of tortuous vessels at hilus In thoracotomy and cVATS groups,the operative time were ( 207.6 ± 88.5 ) vs.( 168.7 ± 55.9 ) min ( P =0.041 ),the blood loss were ( 522.0 ±644.2) vs.(210.1 ± 213.1 ) ml ( P =0.009),the mean chest tube duration were ( 5.4 ± 4.4) vs.(6.3 ± 3.4 ) days ( P ＞0.05 ) and the mean length of hospitalization were ( 10.2 ±4.7 ) vs.( 8.5 ± 3.5 ) days ( P ＞ 0.05 ).No mortality or severe complication occurred in both groups.The morbidity was 25.7％ (9/25)vs.17.1％ (6/35) in thoracotomy and cVATS group,with no significant difference statistically (P =0.133 ) . There were 52％ vs.62.9％ patients achieved symptomatic completely relief and significant improvement was obtained in 40.0％ vs.31.4％ patients in thoracotomy and cVATS group separately.Conclusion cVATS lobectomy is safe and effective in the management of bronchiectasis.

Objective To determine the value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for the diagnosis of intrapulmonary tumors located adjacent to the central airway.MethodsThe study was retrospective,from September 2009 to September 2011,33 patients with pulmonary masses located close to the central airways suspected to be lung cancer were accessed by EBUS-TBNA.Conventional bronchoscopic biopsy before EBUSTBNA was nondiagnostic in all cases.If EBUS-TBNA did not result in a formal pathological diagnosis of malignancy,patients were subsequently referred for a surgical procedure.ResultsOf the 33 patients,EBUS-TBNA confirmed lung cancer in 29 cases (4 small cell lung cancer,25 non-small cell lung cancer).Four patients were not confirmed by EBUS-TBNA,3 cases were diagnosed as squamous cell carcinoma by thoracoscoopy or thoracotomy,the other one was a pulmonary inflammatory lesion diagnosed by thoracoscopy.The sensitivity,specificity,accuracy,negative predictive value and positive predictive value of EBUS-TBNA for the diagnosis of intrapulmonary lesions was 90.2％,100％,90.9％,25％,and 100％,respectively.The procedure was uneventful,and there were no complications.ConclusionEBUS-TBNA is an effective tool with a high yield for the diagnosis of intrapulmonary lesions located adjacent to the central airway.