Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the racliosensitivity of CNE2-implanted tumors, and to investigate the related mechanism. Methods: E1A gene was transfected into CNE2 cells using adenovirus system, and sta-ble E1A positive clones were established. The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was ob-served in nude mice. The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated. The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR. Results: CNE2 cells stably transfected with E1A gene (CNE2-Ad-E1A) were successfully established. The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell- and CNE2-Ad-β-gal cell-implanted mice (CNE2 cells stably transfected with Ad-β-gal). Radiotherapy, E1A gene therapy and E1A gene + radio-therapy all suppressed the growth of implanted tumors, with the tumor suppression rates being (60.32±5.34) %, (70.53±6.12) %, and (97.15±4.87) % , respectively. E1A gene therapy significantly increased the expression of P53 gene in tumor tissues. Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells, and enhance its sensitivity to radiotherapy, which may be related to the increased expression of P53 gene in tumor tissues.

Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the radiosensitivity of CNE2-implanted tumors,and to investigate the related mechanism.Methods: E1A gene was transfected into CNE2 cells using adenovirus system,and stable E1A positive clones were established.The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was observed in nude mice.The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated.The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR.Results: CNE2 cells stably transfected with E1A gene(CNE2-Ad-E1A)were successfully established.The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell-and CNE2-Ad-?-gal cell-implanted mice(CNE2 cells stably transfected with Ad-?-gal).Radiotherapy,E1A gene therapy and E1A gene+radiotherapy all suppressed the growth of implanted tumors,with the tumor suppression rates being(60.32?5.34)%,(70.53?6.12)%,and(97.15?4.87)%,respectively.E1A gene therapy significantly increased the expression of P53 gene in tumor tissues.Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells,and enhance its sensitivity to radiotherapy,which may be related to the increased expression of P53 gene in tumor tissues.

Objective To investigate the effect of El A gene on the radiosensitivity of human laryngeal carcinoma cells and its correlated mechanisms. Methods The Ad-E1A and Ad-β-gal were amplifieated in Hek293 cells, extracted by freezing (-80℃) and thawing(37℃) repeatedly (3 times) , purificated by the method of density gradient of CsC1 and titrated by plaque assay method. Then they were transfected into human laryngeal carcinoma cells (Hep-2) and authenticated by RT-PCR. The radiosensitivity of Hep-2 cells transfeeted with or without El A were studied by cell surviral curve. Finally we investigated the correlated mechanisms including cell apoptosis studied by flow cytometry and VEGF content studied by RT-PCR. Resuits The radiosensitivity of Hep-2 cells transfected with El A was intensified, Do and Dq were lowered and α was increased. Flow cytometry showed that the apoptosis rate of cells with E1A or with El A and radiotherapy was increased. The VEGF content of the cells transfeeted with E1 A or treated by radiotherapy was decreased, which reached the lowest level when the cells were treated with the both mathods. Conclusions E1 A gene can intensify the radiosensitivity and contribute to the apoptosis of human laryngeal carcinoma cells. El A gene and radiotherapy can markedly decrease the VEGF content.

Objective To investigate the clinical and pathological features of microscopic thymoma(MT).Methods The histopathological features of 12 cases of MT were observed by histopathologic and immunohistochemical methods.The pathological morphology,diagnosis,and differential diagnosis were discussed combined with literature.Results 6 cases of MT were accompanied by myasthenia gravis(MG) symptoms.Focal hyperplastic thymic epithelial islands were accompanied by large tracts of mature adipose tissue in 12 cases of MTs,and immunohistochemistry showed CK positive.Cyst formation was found in 5 cases,lymphoid hyperplasia in 7 cases,and vascular proliferation in 5 cases.Conclusions MG may be the clinical manifestation of MT.MT can occur in thymic cortex,medulla and cortex and medulla junction.Since the tumor is small and the lesions are multiple,it can not be found by X-ray or CT examination.Diagnosis depends on histopathological examination.Correct understanding of the clinical and pathological features of MT has guiding significance on the treatment and prognosis judgment of MT.Thymic resection was recommended for MG patients either with or without thymoma.

Objective To explore the effects of different doses of 5,2′,4′-trihydroxy-6,7,5′-trimethoxyflavone nanoparticles (TTF1-NP)on the apoptosis of human hepatoma cells and human normal hepatocytes,and to explore their mechanisms through endoplasmic reticulum stress (ERS)-meditated apoptosis pathway. Methods The human hepatoma cell lines (HepG2,Hep3B and PLC/PRF/5)and human hepatocytes (Chang Liver)were used as cell model, and divided into vehicle, 5-Fu and TTF1-NP treated groups with the concentrations of 50, 100 and 200 μmol·L-1 respectively. The inhibitory effects of TTF1-NP on the cell growth were assessed using MTT assay and the best inhibitory one (HepG-2)was selected as the main research cell lines.Flow cytometry was used to detect the TTF1-NP-induced apoptosis;Western blotting and immunocytochemistry were used to determine the expressions of ERS key proteins.Finally,the expressions of key proteins were detected by Western blotting after using the ERS inhibitor 4-PBA.Results Compared with vehicle group,the inhibitory rates of growth of 4 kinds of human hepatoma cells in different concentrations of TTF1-NP groups were increased (P <0.05 or P <0.01);moreover,the inhibitory effects of TTF1-NP were in a time-and dose-dependent manner.Compared with vehicle group,the apoptotic rates of the cells in TTF1-NP groups were increased in a dose-dependent manner (P <0.05 or P < 0.01 );the expression levels of ERS key proteins GRP78 and caspase-4 were increased with the increasing of the concentration of TTF1-NP (P < 0.05 or P < 0.01).The expression levels of ERS key proteins GRP78 and caspase-4 induced by TTF1-NP were inhibited by ERS inhibitor 4-PBA (P < 0.05 or P < 0.01 ). Conclusion TTF1-NP can induce the apoptosis of HepG2 cells;ERS pathway plays a central role in TTF1-NP-induced apoptosis of HepG-2 cells.

Objective To analyse the relationship between lymphatic vessels invasion and clinical pathological features of papillary thyroid carcinoma ( PTC ) .Methods The expressions of D2-40 and CK19 were examined in the 104 specimens of PTC using immunohistochemical staining with combined monoclonal antibodies and cocktail double enzyme labeled antibody( D2-40/CK19) stainings.The two methods were compared in the diagnosis of PTC metastasis, and the factors affecting lymphatic vessels formation were analyzed.Results The positive rate of lymphatic vessels invasion was 37.5%(39/104) by using immunohistochemical staining with combined monoclonal antibodies and 53.8%( 56/104 ) by cocktail double enzyme labeled antibody ( D2-40/CK19 ) staining ( P<0.05).The lymph node metastasis rate was 83.9%(47/56) in the group with lymphatic vessels invasion, significantly higher than that without invasion 22.9%(11/48, P<0.01).The age of patients, diameter of primary tumor were the influence factors of lymphatic vessels invasion in PTC patients(P<0.05 and P=0.063).Conclusion Cocktail double enzyme labeled antibody ( D2-40/CK19 ) staining is a better method to detect lymphatic vessels invasion in PTC than immunohistochemical staining with combined monoclonal antibodies.

Objective To study the effect of E1A gene on the radiosensitivity of nasopharyngeal carcinoma (NPC) cells and its mechanism. Methods Ad-E1A gene was transfected into human NPC cells (CNE2), then the positive clones (CNE2-Ad-E1A) were identified by RT-PCR. CNE2 cells, CNE2 cells transfected with Ad-β-gal (CNE2-Ad-β-gal) and CNE2-Ad-E1A cells were irradiated with 0 Gy,2 Gy,4 Gy,6 Gy and 8 Gy respectively using 6 MV X-ray. Clone forming assays were carried out, cell survival curves were drawn and the sensitivity enhancing ratio (SER) was calculated. The redistributions of cell cy-cle were analyzed by flow cytometry. RT-PCR was used to detect the expression of wtp53. Results RT-PCR confirmed that E1A gene had been integTated into positively transfected cells and stably expressed. Cell survival curves showed that the SER of D0,Dq and SF_2 value was 1.37, 1.95 and 1.46 in CNE2-Ad-E1A cells. The D_0,D_q and SF_2 value was 1.57 Gy,1.82 Gy, 0.89 in CNE-2 cells and 1.53 Gy,1.78 Gy,0.82 in CNE2-Ad-β-gal cells, respectively. The G_2/M arrest was shown in CNE2-Ad-E1A cells. Moreover, the expression of wtp53 gene was markedly enhanced in Ad-E1A-CNE2 cells. Conclusions E1A gene can ef-fectively enhance the radiosensitivity of human NPC cells, which may be associated the enhancement of wt-p53 expression and G_2/M arrest.

Objective To study the distribution patterns of bacterial flora in sigmoid colon tissues and stools in normal population.Method Bacterial flora were identified and analyzed by using 16sDNA sequencing technology in fresh stool samples (n =13) and colon mucosa samples (n =10).Results The diversity and abundance of bacterial flora were significantly larger in the stool samples than in the sigmoid colon samples (P ＜ 0.001,P ＜ 0.001,P =0.042,P =0.006).The consititution of phylum flora between the two groups were same,including flrmicutes,bacteroides,proteobacteria,and actinomycetes.However,the proportions of firmicutes and bacteroides in stool samples were significantly higher than in the sigmoid colon samples,whereas the proportion of proteobacteria was significantly lower (P ＜ 0.001,P =0.025,P ＜ 0.001).At the genus level,faecalibacterium and bacteroides were the dominant flora in feces,whereas pseudomonas,lactococcus,acinetobacter,and flavobacterium were the most common flora in sigmoid colon mucosa.The amounts of bifidobacterium and lactobacillus were low in both two groups.Conclusion The distribution of bacterial flora remarkably differ in stools and sigmoid colon mucosa.

The aim of this study was to investigate the effect of Paris saponin I (PS I) on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was monitored by the MTT cell viability assay, while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained cells. Western blotting was used to examine the expression of several cell cycle proteins, including cyclin B1 and Cdk1, and the apoptosis-regulated proteins Bcl-2, Bax, cytochrome c, procaspase-9, and procaspase-3. The MTT assay demonstrated that PS I could induce significant dose- and time-dependent inhibition of SGC7901 cell proliferation. Marked morphological changes, including condensation of chromatin, nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining. PSI treatment also resulted in the disruption of the cell cycle at G(2)/M and the induction of apoptosis. Following PSI treatment, the cell cycle-related proteins cyclin B1 and Cdk1 were down-regulated. Expression of the pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 decreased. PSI treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3. These data indicate that PS acts as an inhibitor of proli I feration in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis. PSI is a potential therapeutic agent against human gastric carcinoma.

Objective To investigate the relationship between KRAS,NRAS and BRAF gene mutations and clinicopathological parameters in patients with colorectal carcinoma (CRC).Methods By using TagMan real-time PCR method KRAS/NRAS/BRAF hotspot mutations were detected in 260 cases of CRC.The associations between KRAS/NRAS/BRAF mutation status and clinical pathological characteristics were analysed in different groups divided by gender,age,tumor size,tumor differentiation.Results (1) The KRAS hotspot mutations were G12D,G12A,G12R,G12C,G12V,G12S in codon 12 and G13 C,G13D in codon 13.They were identified in 43.1％ CRC.KRAS mutation rate was higher in females than in males (P =0.05) and the mutation rate in patients ≥ 60 years was significantly higher than that in patients ＜ 60 years(P =0.008).The incidence of metastasis and mortality were higher in KRAS mutant than in KRAS wild type (P =0.004,P =0.037).(2)The NRAS hotspot mutations were in codon1 2,13 and 61.They were identified in 4.6％ CRC.NRAS mutation rate was significantly higher in patients ≥ 60 years and well-differentiated tumors (P =0.032,P =0.042).(3) The mutation rate of BRAF V600E in CRC patients was 4.6％.BRAF V600E mutation rate was significantly higher in patients ≥60 years,with distant metastases and tumors ＞ 5 cm (P =0.032,P =0.026,P =0.038).The incidence of metastasis and rucurrence and mortality were higher in BRAF mutant (P =0.030,P =0.002,P =0.007).Conclusions In CRC patients,KRAS mutations correlate with demographic factors,metastasis and mortality,NRAS mutations correlate with age and tumor differentiation,while BRAF mutation correlate with age,tumor size,metastasis,recurrence and mortality.