Objective To quantitatively compare the γ-H2AX foci formation between DNA-PKcs+/+and DNA-PKcs-/-mouse embryonic fibroblast(MEF)cells,and to investigate the dynamic changes in DNA double-strand breaks(DSBs)in human nasopharyngeal carcinoma SUNE-1 cells exposed to X-ray radiation. Methods The expression of DNA-PKcs was determined by Western blot. The γ-H2AX foci formation induced by 5 Gy X-ray radiation was detected by cell immunofluorescence. The ImageJ software was used to quantitatively analyze the γ-H2AX foci formation. Results The expression of DNA-PKcs was silenced in DNA-PKcs-/-MEF cells and normal in DNA-PKcs+/+MEF cells. According to the dynamic analyses of the numbers of γ-H2AX foci/cell and γ-H2AX foci/mm2, a similar tendency was observed in DSB formation in DNA-PKcs+/+MEF cells, DNA-PKcs-/-MEF cells,and SUNE-1 cells exposed to X-ray radiation. A large number of γ-H2AX foci formed at 0.5-1.0 h after radiation. DSBs were repaired at 6 h after radiation in DNA-PKcs+/+MEF cells and 24 h after radiation in DNA-PKcs-/-MEF cells and SUNE-1 cells. The peak values of γ-H2AX foci/cell and γ-H2AX foci/mm2were observed at 1.0 and 0.5 h after radiation, respectively. Compared with DNA-PKcs+/+MEF cells, DNA-PKcs-/-MEF cells had different numbers of γ-H2AX foci/cell at 0.5, 1.0, 3.0, 6.0, and 12.0 h after radiation, as well as different numbers of γ-H2AX foci/mm2at 3.0, 6.0, and 12.0 h after radiation. Conclusions Quantitative measurement of the number of γ-H2AX foci/cell or γ-H2AX foci/mm2by cell immunofluorescence provides new insights into the quantitative and dynamic study of DSB damage and repair.

OBJECTIVE: To determine clinical features and diagnostic methods for primary tracheobronchial amyloidosis (TBA).
 METHODS: The clinical manifestations and diagnosis of a male patient who had been misdiagnosed for many years were described and analyzed.
 RESULTS: The patient was a 68-year-old male who complained of recurrent cough, expectoration, and progressive dyspnea for more than 30 years. He had been diagnosed with chronic bronchitis, bronchiectasis, and endobronchial tuberculosis in other hospitals and treated with antibiotics frequently and anti-tubercular agents for 3 months. Despite the treatments, the patient's symptoms were progressively worse. Finally, he came to Xiangya Hospital, Central South University, and was clearly diagnosed with primary TBA based on histopathological evidence after bronchoscopy.
 CONCLUSION TBA, a rare disease resulting from abnormal submucosal amyloid deposition in the trachea and bronchi, may display with many different symptoms. TBA is often misdiagnosed with other pulmonary diseases. The use of bronchoscopic techniques is essential for the diagnosis of TBA. Histopathology remains the gold standard for diagnosis of primary TBA. So, for patients with chronic cough of unknown etiology, bronchoscopy should be performed to obtain biopsy samples for the definitive diagnosis.
Aged ; Amyloidosis ; diagnosis ; Bronchi ; pathology ; Bronchial Diseases ; diagnosis ; Bronchiectasis ; Bronchitis, Chronic ; Bronchoscopy ; Delayed Diagnosis ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Male ; Trachea ; pathology ; Tracheal Diseases ; diagnosis ; Tuberculosis

OBJECTIVE: To establish a stable A549 cell line transfected by RNA binding motif 5 (RBM5) expression vector, and to investigate the effect of RBM5 gene on proliferation of A549 cell line and the expression of DEAH box polypeptide 15 (DHX15). METHODS: The eukaryotic expression vector pcDNA3.1 (+)/RBM5 was constructed by a twostep PCR technique. Then, the recombinant plasmid pcDNA3.1 (+)/RBM5 was verified by DNA sequencing and transfected into the lung adenocarcinoma cell A549. The positive cells with overexpression of RBM5 gene were identified by Western blotting. Flow cytometry was used to analyze the cell cycles of the positive A549 cells [pcDNA3.1 (+)/RBM5-A549] and the negative controls [pcDNA3 .1 (+)- A549]. Finally, RT-PCR was used to detect the expression of DHX15, a splicing-related factor, in the positively transfected A549 cells and the negative controls. RESULTS: A pcDNA3.1 (+)/RBM5 eukaryotic expression vector has been constructed successfully, and the A549 cell line that stably transfected with RBM5 gene has been established. Compared with negative control cells, the percentage of G1 phase cells in the positive cells was increased, while the percentage of S phase was decreased (both P<0.01), and the expression of DHX15 is upregulated (P<0.01). CONCLUSION RBM5 gene can inhibit the cell cycle and upregulate the expression of DHX15 in A549 cells.
Cell Cycle ; Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Cell Proliferation ; DNA-Binding Proteins ; genetics ; Genetic Vectors ; Humans ; RNA Helicases ; metabolism ; RNA-Binding Proteins ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics

Objective To investigate the maximum-tolerated dose (MTD) of cisplatin in docetaxel,cisplatin,and fluorouracil (TPF) induction chemotherapy followed by intensity-modulated radiotherapy (IMRT) and concomitant chemotherapy as well as the safety and short-term efficacy of TPF induction chemotherapy in the treatment of locally advanced nasopharyngeal carcinoma (NPC).Methods Thirtythree patients with locally advanced NPC were enrolled in this trial.The MTD of cisplatin was determined by dose escalation study,and the short-term efficacy and toxicities were evaluated.Results When the doses of docetaxel and fluorouracil were 60 mg/m2 d1 and 550 mg/m2 d1-5,respectively,the MTD of cisplatin was 65 mg/m2 d1.In this regimen (repeated every 3 weeks),grade 3-4 toxicities included neutropenia (67％),febrile neutropenia (9％),diarrhea (21％),and oral mucositis (6％).Except those who experienced dose-limited toxicity,other patients completed the whole treatment schedule.After TPF induction chemotherapy,the overall response rate was 97％,and the complete response rate was 21％.Conclusions In the endemic areas of NPC,induction chemotherapy with docetaxel (60 mg/m2 d1),cisplatin (65 mg/m2 d1),and fluorouracil (550 mg/m2 d1-5),which is repeated every 3 weeks,is proved safe and effective for Asian patients with locally advanced NPC.

Objective To approach the effects of MMP-9 antisensc oligonucleotide(ASODN)on human lung adcnocarcinoma(A549) cell apoptosis and proliferation capability.Methods MTT.was used to analyze the effect that MMP-9 transfection on A549 cell growth.Flow cytometry was used to analyze the ratio of cell proliferation and apoptosis.RT-PCR WaS used to detect the expression of MMP-9mRNA and Western blot was used to detect the changes of MMP-9 protein.Results In some degree,MMP-9 ASODN that inhibited the survival rate of the A549 cell presented concentration and time dependence manner.The best dependence concentration of ASODN was 600nmol/L for 24 hour.After trasnfection of MMP-9ASODN to A549 cell.the perceive of A549 apoptosis cell was significandy higher than that in control group(P＜0.01).When the antisense oligonucleotides concentration is 600nmol/L and the action time is 48 hours,the relative expression level of MMP-9mRNA and MMP-9 protein are obviously less than that in control group(P＜0.01).Conclusion MMP-9ASODN may down regulate the expression of MMP-9mRNA and MMP-9 protein,effectively inhibit the proliferation of A549 cell and promote the apoptosis of A549 cell.

BACKGROUNDDecorin is a member of the small proteglycans in extracellular matrix of tumor microenvironment, which is known to relate to the initiation, progression and growth of the tumor. The aim of this study is to investigate the effects and mechanism of decorin on the proliferation of A549 lung adenocarcinoma cell line in vitro.

METHODSLung adenocarcinoma cell line A549 was cultured with decorin in a wide range of concentration for different time. Cell activities were studied by MTT. The changes of cell cycle and apoptosis were analyzed by FCM. Decorin mRNA expression was detected by RT-PCR. P21 expression was determined by Western blot. TGF-β concentration in the culture supernatants was determined by ELISA.

RESULTSThe proliferation of A549 cell could be inhibited by decorin in vitro and the inhibition effect was the time- and dose-dependent relationship. Apoptosis of adenocarcinoma cell could be efficiently induced by decorin in a time/dose-dependent manner. Decorin could upregulate the intrinsic decorin mRNA and P21 protein expression, downregulate the TGF-β, and block cell cycle at G1 phase.

CONCLUSIONSDecorin can inhibit adenocarcinoma cell proliferation and induce apoptosis of adenocarcinoma cells in vitro. The proliferation of A549 cell could be inhibited in vitro by decorin through the mechanism of increasing decorin mRNA, decreasing TGF-β, increasing P21 protein expression, inhibiting cell cycle and inducing cell apoptosis.

BACKGROUNDMatrix metalloproteinase-9 (MMP-9) is one of significant extracellular matrix catabolic enzymes, which is known to relate to the initiation, progression and growth of the tumor. The aim of this study is to investigate the effects of MMP-9 antisense oligonucleotide (ASODN) on the apoptosis and metastasis of human lung adenocarcinoma A549 cell.

METHODSMTT was used to analyze the effect of MMP-9 ASODN transfection on A549 cell growth. Flow cytometry was used to analyze the cell cycle and apoptosis. MTT chromometry and scratch migration were used to observe the capability of adhesion and immigration of the A549 cell.

RESULTSMMP-9 ASODN inhibited the survival rate of the A549 cell and the action was concentration-and time-dependent. The peak concentration of MMP-9 ASODN was 600 nmol/L at 48 hours after transfection. After transfection of MMP-9 ASODN to A549 cell, the percentage of A549 apoptosis cell was significantly higher than that of control group (P < 0.01), but the adhesion and migration cut down predominantly (P < 0.01).

CONCLUSIONSMMP-9 ASODN can depress the adhesion and migration and inhibit proliferation of adenocarcinoma A549 cell effectively, and can promote apoptosis of the A549 cell.

Objective: To investigate effects of lorsartan, fosinopril on myocardial fibrosis, angiotensin Ⅱ and cardiac remolding in the spontaneously hypertensive rats (SHR). Methods: 16-week-old SHRs were divided randomly into 3 groups: SHR-L (treated with lorsartan), SHR-F (treated with fosinopril) and SHR-C (untreated), each group consisting of 10 rats. After 8 weeks' and 16 weeks' therapeutic period, collagen volume fraction (CVF), perivascular circuferential area (PVCA), plasma and myocardium angiotensin Ⅱ concentrations were examined by pathological examination with computed processing and radioimmunoassay respectively. Results: (1) Compared with SHR-C after 8 weeks' and 16 weeks' therapeutic period, the systolic blood pressure (SBP) was decreased similarly in both treatment groups. Heart and left ventricular weights, heart weight and eft ventricular mass indexes were lower significantly in both treatment groups than in SHR-C. Left ventricular mass index was reduced to a lower extent in SHR-F group than in SHR-L group after 16 weeks. (2) Compared with SHR-C, CVF, PVCA after 8 weeks and 16 weeks were reduced significantly in SHR-F and SHR-L. Meanwhile, CVF after 16 weeks in SHR-F than in SHR-L. (3) Compared with SHR-C after both therapeutic periods, plasma and myocardium angiotensin Ⅱ concentrations were increased Significantly in SHR-L, but plasma angiotensin Ⅱ concentrations were not altered significantly in SHR-F. However, myocardium angiotensin Ⅱ concentrations were reduced significantly in SHR-F after 8 weeks and 16 weeks in SHR-F. Conclusion: Lorsartan, fosinopril inhibit myocardial fibrosis and reverse heart hypertrophy. Fosinopril may be more effective in these above effects than Lorsartan. The mechanism of the both drug's cardioprotective effects was related to inhibition of myocardium rennin-angiotension-aldsteron system.

Objective To explore the effect of glucocorticoid on glucose transport activity and the expression of insulin signaling peptides in the primary cultured rat adipocytes .Methods Isolated rat adipocytes were cultured for 2h,8h,16h and 24h respectively at 5 mmol glucose with dexamethasone (Dex) 0 3?mol. Then the glucose uptake, cellular contents of insulin receptor substrate (IRS) 1/2, phosphatidylinositol 3-kinase 85 subunit (p85) and protein kinase B(PKB)were measured by Western blotting.Results These adipocytes treated with Dex have shown to impair the basal and insulin-induced glucose uptake with a maximum inhibition at 2h and 24h respectively; Dex down-regulated IRS1 protein expression in a time-dependent manner and up-regulated IRS2 content. The cellular p85 and PKB contents were also decreased by Dex. Conclusion Chronic exposure to glucocorticoid can inhibit glucose uptake and induce insulin resistance. The mechanism may be involved in affecting the expression of insulin signaling peptides.

Objective To understand current situation of live quality of old patients and to explore its influencing factors.Methods Age of all subjects more than 60 years who was aged in-patients with a chronic disease in geriatric ward,department of XiangYa Hospital. General physical examination were executed. Symptom Checklist 90(SCL-90) and Quality of Life (QOL) scales were measured.Results ⑴The problems of QOL and mentality were occurred in 78%,77 33% respectively of the patients. ⑵Multiple linear regression showed that financial condition, mentality, age and physical disease were major factors that affected the patients' quality of life . The linear equation was YQOL=36 967-0 734 X age+2 012 X financial-0 0352 X mentality-0 712 X diseases(P