Delightful achievements have been obtained in forestry genetic breeding since the application of transgenic technology in this field during the past 20 years. Field trials of some genetic modified (GM) trees have been carried out, and some GM trees have been commercialized. Meanwhile, the risks of ecological safety caused by GM trees have raised attention in the public gradually. These issues mainly include the horizontal transfer and vertical flow of foreign genes, and the potential effects on insects, soil ecosystems and virus. The current status of field trials, commercial applications and the potential ecological risks of GM trees were summarized. Then the prospects of GM trees were also presented.

OBJECTIVETo evaluate the activity of DNA polbeta promoter on p53 gene in salivary adenoid cystic carcinoma (SACC) cells.

METHODSThe luciferase activity was examined and used to evaluate the activity of DNA polbeta promoter on SACC-83 cells. Eukaryotic expression plasmids of p53 gene were constructed and stably transfected into SACC-83 cells. RT-PCR was used to assess the expression of p53 gene. The SACC-83 cells were subjected to the treatments of H2O2, ultraviolet radiation, Bleocin, and affected p53 mRNA and protein level in SACC-83 cells were characterized with RT-PCR and Western blotting.

RESULTSThe result of luciferase activity proved that the activity of DNA polbeta promoter in SACC-83 cells was much higher than that of CMV promoter. The results of RT-PCR suggested that p53 gene with different promoters were all expressed effectively, but the expression efficiency was different. It was greater in DNA polbeta group than in CMV group. After DNA damage, p53 gene expression increased and DNA polbeta promoter could enhance the expression of p53 gene more than CMV promoter. The results of Western blotting indicated that the expression of P53 protein between the two groups did not show any difference.

CONCLUSIONIn SACC cells, the activity of DNA polbeta promoter was increased and DNA polbeta promoter could enhance the expression of p53.

Carcinoma, Adenoid Cystic ; Cell Line, Tumor ; DNA ; Genes, p53 ; Humans ; Hydrogen Peroxide ; Promoter Regions, Genetic ; RNA, Messenger ; Salivary Gland Neoplasms ; Transfection ; Ultraviolet Rays

OBJECTIVETo establish a model of ER stress-induced apoptosis with tunicamycin and to examine whether Bim is dysregulated and its potential role in resistance of melanoma cells to apoptosis under endoplasmic reticulum (ER) stress.

METHODSA model of ER stress-induced apoptosis was established with tunicamycin. Apoptotic cells were quantitated using the annexin V/propidium iodide method by flow cytometry. Hoechst staining was also used to confirm the apoptotic cell death. Western blotting was used to measure the activation of caspase-3 and -9, and the expression of Bim, GRP78, CHOP, and Foxo1 at the protein level. The expression of Bim, CHOP and Foxo1 at the mRNA level was quantitated by qPCR. The siRNA technique was used to inhibit the expression of Bim.

RESULTSTreatment of the melanoma cells with tunicamycin did not induce significant apoptosis and activation of caspase cascade, whereas it caused marked activation of caspase-3 and -9, and apoptosis in HEK293 cells which were used as a control. With exposure to tunicamycin (3 µmol/L) for 12, 24, 36 hours the Bim protein levels were not increased in Mel-RM and MM200 cells. Its mRNA levels were 0.37 ± 0.05, 0.13 ± 0.02 and 0.02 ± 0.01 in Mel-RM cells, while 0.41 ± 0.06, 0.16 ± 0.04 and 0.21 ± 0.03 in MM200 cells, respectively. The expression of Bim mRNA was significantly reduced compared with that in the control groups of the two cell lines (P < 0.01). siRNA knockdown of Bim protected HEK293 cells against activation of caspase-3. The cell apoptosis of Bim siRNA group was (5.69 ± 0.38)%, significantly lower than that of the siRNA control group (40.32 ± 1.64)% and blank control group (35.46 ± 2.01)% (P < 0.01). In the melanoma cells after exposure to tunicamycin (3 µmol/L) for 6, 12, 24, and 36 hours the transcription factor CHOP at mRNA level were significantly increased and the expressions at protein level were also up-regulated. The expressions of another transcription factor Foxo1 at mRNA level significantly decreased and the expressions at protein level were down-regulated, too.

CONCLUSIONSThe lack of Bim up-regulation contributes to the resistance of melanoma cells to ER stress-induced apoptosis and may be a mechanism by which melanoma cells adapt to ER stress conditions. Transcription factors CHOP and Foxo1 may be responsible for the dysregulation of Bim in melanoma cells upon ER stress.

Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Bcl-2-Like Protein 11 ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Endoplasmic Reticulum Stress ; drug effects ; Forkhead Box Protein O1 ; Forkhead Transcription Factors ; genetics ; metabolism ; HEK293 Cells ; Heat-Shock Proteins ; metabolism ; Humans ; Melanoma ; genetics ; metabolism ; pathology ; Membrane Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transcription Factor CHOP ; genetics ; metabolism ; Tunicamycin ; pharmacology

OBJECTIVETo study the oncogene transcriptor c-myc, stimulatory protein 1 (SP1) expression in ameloblastoma (AB) and their relation with telomerase reverse transcripase (hTERT), and to investigate the clinical biological characteristics of AB.

METHODSThe expression was observed in AB by in situ hybridization and SP method.

RESULTSThe positive rates of c-myc mRNA, hTERT mRNA and SP1 protein were 81.5% (44/54), 94.4% (51/54) and 83.3% (45/54), respectively. Their positive rates increased as AB recurred and transformed malignantly. A strong correlation was found between hTERT and c-myc, hTERT and SP1 (rs = 0.853, P < 0.001; rs = 0.900, P < 0.001).

CONCLUSIONActivity of telomerase plays an important role in the tumorigenesis development of AB. Increasing of hTERT expression may be related to c-myc and SP1. The expression of these three parameters has a significant correlation with the clinical biological characteristics of AB.

Ameloblastoma ; metabolism ; Humans ; Proto-Oncogene Proteins c-myc ; metabolism ; Sp1 Transcription Factor ; metabolism ; Telomerase ; metabolism

OBJECTIVETo study the expression of HOXC13 mRNA in ameloblastoma (AB), and to investigate its biological significance.

METHODSHOXC13 mRNA was examined in 47 cases of AB (primary AB 29 cases, recurrent AB 14 cases, malignant AB 4 cases). 2 cases of fibrous dysplasia of bone, 10 cases of keratocystic odontogenic tumor (KCOT) and 7 cases of normal oral mucosa were selected as control.

RESULTSThe positive rates of HOXC13 mRNA in AB, KCOT, and normal oral mucosa were 97.9% (46/47), 7/10 and 3/7, respectively. There was a significant difference among AB, OKC and normal mucosa (chi(2) = 21.665, P = 0.001). For HOXC13, the keratinizing cells and granulizing cells in AB were negative, some fibroblasts were positive, 2 cases of fibrous dysplasia of bone were positive.

CONCLUSIONSHOXC13 was highly expressed in AB. The expression of HOXC13 mRNA in AB had heterogeneity, which could improve the epithelial proliferation, and its loss may lead to the cornification and degeneration of epithelial cells.

Adolescent ; Adult ; Ameloblastoma ; genetics ; metabolism ; pathology ; Female ; Gene Expression ; Genes, Homeobox ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; Odontogenic Tumors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Young Adult

AIM: To explore the role of PI3K/Akt/nNOS in Zhouluotong extract resisting diabetic peripheral neuropathy.METHODS:The Schwann cells were divided into normal group ( D-glucose 25 mmol/L) , model group ( D-glucose 100 mmol/L) , Zhouluotong extract Z-6 +high glucose group, Zhouluotong +high glucose group, mecobalamine+high glucose group.The viability, nitric oxide content and the Ca2+-ATPase activity in Schwann cells were determined by Cell Counting Kit-8 , nitric oxide assay kit and Ca2+-ATPase assay kit, respectively.The apoptosis of Schwann cells were analyzed by flow cytometry.The expression of Bcl-2, Bcl-xL, Bax, Bak and caspase-3, and the phosphorylation levels of nNOS and Akt were determined by Western blotting.The signal pathway of PI3K/Akt was explored by dominant negative PI3K and Akt (δp85 and DN-Akt) transient transfection assay.RESULTS:Under high-glucose culture, the cell viability, nitric oxide content in culture supernatant, the expression of Bcl-2 and Bcl-xL, and the phosphorylation levels of Akt and nNOS in the Schwann cells were significantly increased.The cell apoptosis, the expression of Bax, Bak and caspase 3 in the Schwann cells were significantly decreased by Zhouluotong extract Z-6, compared with model group.In-creased nitric oxide content and the up-regulation of nNOS were observed.However, the effects of blocking PI3K/Akt, the upstream pathway of nNOS , by transfection with DN-δp85 on Akt phosphorylation in the Schwann cells was still unclear. CONCLUSION:Zhouluotong extract Z-6 changes the phosphorylation of nNOS, and the expression of anti-apoptotic fac-tors , caspase-3 and pro-apoptotic factors in Schwann cells under high-glucose culture, thus reducing apoptosis and elevating viability.The relationship to PI3K/Akt/nNOS pathway needs further investigation.

Objective To evaluate the reliability of HARSHAW-3500 thermoluminescence dosimetry system by testing its performances.Methods HARSHAW-3500 thermoluminescence dosimetry system had its performances tested and evaluated according to Verification regulation of thermoluminescence dosimetry systems used in persontal and environmental monitoring forXandgammaradiation(JJG 593-2006),Testingcriteriaofpersonneldosimetryperformanceforexternalexposure (GBZ 207-2016),Specifications for individual monitoring of occupational external exposure (GBZ 128-2016) and Thermoluminescence dosimetry systems for personal and environmental monitoring (GB/T 10264-2014),such as batch homogeneity,repeatability,linearity,incidence angle response,stability,energy response and scale factor,quantity inspection,residual dose,detection limit and etc.Results Testing results of various performance indicators proved to be within the limits according to national and industrial standards.Conclusion HARSHAW-3500 thermoluminescence dosimetry system conforms to the requirements for radiation dose measurement.It is beneficial to the improvement of quality and performance of thermoluminescence dosimetry by performances analysis and evaluation.

AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P ＜ 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P ＜ 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.

OBJECTIVETo evaluate the inhibitory effect of p53 gene on salivary adenoid cystic carcinoma (SACC) cells.

METHODSAdenoviral vector pDeltaE1-p53 was constructed and transfected into SACC-83 cells. The enhanced p53 expression was measured by reverse transcription polymerase chain reaction (RT-PCR), and the effects of transfected p53 on SACC-83 cells were analyzed by TRAP-PCR-ELISA, luciferase reporter, flow cytometry (FCM), soft agar assay and tumorigenicity test.

RESULTSThe expression of p53 gene in SACC-83 cells was increased after introduction of pDeltaE1-p53. The telomerase activity and the transcriptional activity of hTERT promoter were inhibited. The cells cycles of transfected SACC-83 were arrested in G(1) phase and the rate of colony-formation was decreased, and similarly the tumorigenicity in nude mice was also decreased.

CONCLUSIONSThe introduction of wild-type p53 by adenoviral vector could suppress the telomerase activity and malignant phenotypes of salivary adenoid cystic carcinoma cells.

Animals ; Carcinoma, Adenoid Cystic ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Genetic Vectors ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Salivary Gland Neoplasms ; genetics ; metabolism ; pathology ; Telomerase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Protein p53 ; genetics

Objective To demonstrate molecular characterization of a newly isolated enterovirus.Methods Virus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5' ,3'RACE (rapid amplification of eDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus. Results Sequence analysis showed that this newly isolated enterovirus shared 83 %-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89.Phylogenetic analysis indicated that it was probably a new subtype of enterovirus 89. Conclusion This newly isolated enterovirus in the stool specimen from patient has the same serotype with enterovirus 89, but it was probably a new subtype of enterovirus 89.