Delayed callose degradation restores the fertility of multiple P/TGMS lines in Arabidopsis

Photoperiod/temperature-sensitive genic male sterility(P/TGMS)is widely applied for improving crop production.Previous investigations using the reversible male sterile(rvms)mutant showed that slow development is a general mechanism for restoring fertility to P/TGMS lines in Arabidopsis.In this work,we isolated a restorer of rvms–2(res3),as the male sterility of rvms–2 was rescued by res3.Phenotype analysis and molecular cloning show that a point mutation in UPEX1 l in res3 leads to delayed secretion of callase A6 from the tapetum to the locule and tetrad callose wall degradation.Electrophoretic mobility shift assay and chromatin immunoprecipitation analysis demonstrated that the tapetal transcription factor ABORTED MICROSPORES directly regulates UPEX1 expression,revealing a pathway for tapetum secretory function.Early degradation of the callose wall in the transgenic line eliminated the fertility restoration effect of res3.The fertility of multiple known P/TGMS lines with pollen wall defects was also restored by res3.We propose that the remnant callose wall may broadly compensate for the pollen wall defects of P/TGMS lines by providing protection for pollen formation.A cellular mechanism is proposed to explain how slow development restores the fertility of P/TGMS lines in Arabidopsis.

Studies on GA_3 Spraying Dosage for eui TGMS Rice Changxuan 3S in Its Hybrid Seed Production

[Objective] Changxuan 3S was thermo-sensitive genicmale sterile（TGMS）rice selected from irradiated seeds of Peiai 64S by 350 Gy^（60）Coγ-ray.The aim of the study was to confirm GA3 spraying dosage of Changxuan 3S with eui gene in its hybrid seed production.[Method] Changxuan 3S possessing eui gene and its parent Peiai 64S were chosen as materials.Comparison studies on sensitivity to GA3 in their hybrid seed production were carried out.[Result] The suitable stage for spraying GA3 in the hybrid seed production of Changxuan 3S was at 10% of panicles headed;The optimal dosage was 90 g/hm2 with 2 split sprayings,the first spraying of 45 g/hm2 at heading of 10% panicles and the second one of 45 g/hm2 on the following day.Under the condition of spraying GA3 at the rate of 90 g/hm2,the panicle neck exsertions of Changxuan 3S was ＋1.78 cm,and exserted stigma rate and seed setting rate of Changxuan 3S were 96.87% and 36.44%,being 21.46% and 16.33% more than those of Peiai 64S,respectively.The theoretical yield of ＂Changxuan 3S/9311＂ reached 2 931.90 kg/hm2,which was increased by 1 259.40 kg/hm2 comparing with ＂Peiai 64S/9311＂.[Conclusion] Compared with Peiai 64S,Changxuan 3S is more sensitive to GA3,which results in no or little using GA3 in seed production of Changxuan 3S.Moreover,Changxuan 3S showed higher yield potential than Peiai 64S.

18β-glycyrrhetinic acid inhibits proliferation of gastric cancer cells through regulating the miR-345-5p/TGM2 signaling pathway

BACKGROUND Gastric cancer(GC)is a common gastrointestinal malignancy worldwide.Based on cancer-related mortality,the current prevention and treatment strategies for GC still show poor clinical results.Therefore,it is important to find effective drug treatment targets.AIM To explore the molecular mechanism of 18β-glycyrrhetinic acid(18β-GRA)regulating the miR-345-5p/TGM2 signaling pathway to inhibit the proliferation of GC cells.METHODS CCK-8 assay was used to determine the effect of 18β-GRA on the survival rate of GES-1 cells and AGS and HGC-27 cells.Cell cycle and apoptosis were detected by flow cytometry,cell migration was detected by a wound healing assay,the effect of 18β-GRA on subcutaneous tumor growth in BALB/c nude mice was investigated,and the cell autophagy level was determined by MDC staining.TMT proteomic analysis was used to detect the differentially expressed autophagy-related proteins in GC cells after 18β-GRA intervention,and then the protein-protein interaction was predicted using STRING(https://string-db.org/).MicroRNAs(miRNAs)transcriptome analysis was used to detect the miRNA differential expression profile,and use miRBase(https://www.mirbase/)and TargetScan(https://www.targetscan.org/)to predict the miRNA and complementary binding sites.Quantitative real-time polymerase chain reaction was used to detect the expression level of miRNA in 18β-GRA treated cells,and western blot was used to detect the expression of autophagy related proteins.Finally,the effect of miR-345-5p on GC cells was verified by mir-345-5p overexpression.RESULTS 18β-GRA could inhibit GC cells viability,promote cell apoptosis,block cell cycle,reduce cell wound healing ability,and inhibit the GC cells growth in vivo.MDC staining results showed that 18β-GRA could promote autophagy in GC cells.By TMT proteomic analysis and miRNAs transcriptome analysis,it was concluded that 18β-GRA could down-regulate TGM2 expression and up-regulate miR-345-5p expression in GC cells.Subsequently,we verified that TGM2 is the target of miR-345-5p,and that overexpression of miR-345-5p significantly inhibited the protein expression level of TGM2.Western blot showed that the expression of autophagy-related proteins of TGM2 and p62 was significantly reduced,and LC3II,ULK1 and AMPK expression was significantly increased in GC cells treated with 18β-GRA.Overexpression of miR-345-5p not only inhibited the expression of TGM2,but also inhibited the proliferation of GC cells by promoting cell apoptosis and arresting cell cycle.CONCLUSION 18β-GRA inhibits the proliferation of GC cells and promotes autophagy by regulating the miR-345-5p/TGM2 signaling pathway.

基于P-中心模型的城市轨道交通线网应急救援站选址研究

[目的]随着城市轨道交通线网的扩大,其系统风险变得更为复杂且不可控。为提高城市轨道交通系统的应急救援能力,需对线网内应急救援站的选址问题展开研究。[方法]将地铁覆盖区域按照城市道路环线进行分区限速,得到任意路径的速度矩阵,结合距离矩阵进行转换后得到时间矩阵。在此基础上,计算任意站点间基于时间的最短路径。考虑时间、救援关系、成本3个方面的约束条件,建立了以系统最大救援时间最小化为目标的P-中心选址模型。以武汉市城市轨道交通线网应急救援站选址为案例,应用该模型,并采用自适应遗传算法在MATLAB软件中进行编程求解,得出该线网应急救援站选址的最优方案。[结果及结论]与相似研究相比,该选址方案投入更少但效益更大,充分证明了所提的P-中心模型具有先进性。

水稻CMS和TGMS近等基因系的构建及其配合力评价

以温敏核不育系培矮64S和6311S为母本和轮回亲本,与保持系新协黄B杂交,采用回交-测交的方法,排除恢复基因和温敏核不育基因,首次育成了温敏核雄性不育系的质核互作型雄性不育近等基因系培矮64A和6311A。并配组了一套4个不育系×6个恢复系的不完全双列杂交组合。研究了温敏核不育系和质核互作型雄性不育近等基因系培矮64S和培矮64A及6311S和6311A的育性、形态特征和一般配合力的差异。结果表明,在遗传背景相同的前提下,不育期的温敏核不育系与其相应的质核互作型雄性不育系除花粉特征、柱头大小和异交结实率有一定差异外,其他农艺经济性状均表现一致。近等基因系培矮64S和培矮64A与6311S和6311A产量及其他农艺性状的一般配合力存在显著差异,但是培矮64S和培矮64A、6311S和6311A的产量及其他农艺性状的一般配合力没有显著差异,说明水稻杂种优势的高低与两系三系法没有必然的联系。

基于通用可调(p,q)阶Rife-Vincent自乘-卷积窗的改进四谱线插值高精度谐波分析算法

加窗快速傅里叶变换(WIFFT)是一类重要的谐波参数估计方法。然而,快速傅里叶变换固有的栅栏效应以及非同步采样产生的频谱泄漏对谐波参数的估计精度有较大影响。为此,利用具有最大旁瓣衰减特性的I类Rife-Vincent窗作为父窗,提出一种新型的通用可调(p,q)阶Rife-Vincent自乘-卷积窗(RVSMC p-q),其特点在于通过改变乘积阶数和卷积阶数可以灵活地改变窗函数的主瓣宽度、旁瓣峰值和衰减速度。在此基础上,提出一种基于RVSMC p-q窗的四谱线插值高精度谐波参数估计方法,利用最小二乘拟合原理给出谐波信号的幅值、频率及相位参数的低复杂度多项式估计公式。在复杂谐波、间谐波及基波频率波动的情况下对基于RVSMC p-q窗的四谱线插值谐波参数估计方法的效果进行仿真。研究结果表明:在较低的乘积和卷积阶数条件下,新算法可以获得比经典的基于自乘积窗或自卷积窗的WIFFT算法更为优越的参数估计精度,且算法复杂度较低。

Defensive physiological characters of Pyropia yezoensis resistant lines to the red rot disease

To explore ef fective physiological indexes to distinguish the resistant lines and susceptible lines of Pyt hium porphyrae,the causal agent of red rot disease of Pyropia yezoensis,and establish the disease-resistance breeding strategy,we obtained and analyzed the candidate resistant and susceptible lines by population selection.Gametophytes of the candidate lines were cultured in seawater containing Pyt.porphyrae zoospores.Antioxidase activities,including superoxide dismutase(SOD),peroxidase(POD)and polyphenol oxidase(PPO),were measured and compared between the two lines before and after infection.In the resistant lines,SOD and POD activities increased and then decreased,but PPO activity rose with the prolongation of the infection time.The phenylalanine ammonia lyase(PAL)activities also increased and then decreased after infection,but it had significantly different expression in the two lines without pathogen attack.The synthesis rates ofβ-1,3-glucanase,and cell-wall degrading enzyme were different from each other between the two lines after infection of P yt.porphyrae.Comparison in the contents of malondialdehyde(MDA)and reactive oxygen species(ROS)in the two lines showed that,the two contents varied synchronously in response to the pathogen attack.Changes of these enzymes activities or contents demonstrated that Pyr.yezoensis could resist against the pathogen of P yt.porphyrae with the antioxidant defense capacity.In addition,β-1.3-glucanase content showed extremely significant dif ference between the two lines,and the PAL had consistent expression dif ference.Therefore,phenylalanine ammonia lyase(PAL)andβ-1,3-glucanase can be considered as an ef fective index to distinguish susceptible line and resistant line,with which the workload of the resistant breeding could be reduced in the future.

Apoptosis mechanisms of human gastric cancer cell line MKN-45 infected with human mutant p27

AIM: To explore the inducing effect of human mutant p27 gene on the apoptosis of the human gastric cancer cell line MKN-45 and its associated mechanisms. METHODS: The recombinant adenovirus Ad-p27mt was constructed to infect the human gastric cancer cell line MKN-45. Using flow cytometry, TUNEL assay and DNA fragment analysis, we measured the apoptotic effect of Ad-p27mt on the human gastric cancer cells. RESULTS: Ad-p27mt was successfully constructed and the infection efficiency reached 100%. After 18 h of infection, we observed an apoptotic hypodiploid peak on the flow cytometer before G1-S and apoptotic characteristic bands in the DNA electrophoresis. The apoptotic rate detected by TUNEL method was significantly higher in the Ad-p27mt group (89.4±3.12%)compared to the control group (3.12±0.13%, P ＜ 0.01).CONCLUSION: Human mutant p27 can induce apoptosis of the human gastric cancer cells in vitro.

MiR-25-3p attenuates the proliferation of tongue squamous cell carcinoma cell line Tca8113

Objective:To investigate the effects of miR-25-3p on the occurrence,development and proliferation of tongue squamous cell carcinoma cells.Methods:To establish tongue squamous cell carcinoma cell line Tca8113 that stably and highly express miR-25-3p using recombinant reiroviral vector-mediated gene transfer method.The proliferation of transfected Tca8113 was detected by thiazolyl blue tetrazolium bromide(MTT)and cell colony formation assays.eyclnD1,p21^(cipt)and p27^(kipt)mRNA expressions in the transfected Tca-8113 were detected by quantitative PCR.cyclinD1,p21^(cipt),p27^(kipt),AKT,p-AKT,FOXOt and p-FOX01 expressions in the transfected Tca8113 were detected by western blot analysis.In addition,miR-25-3p expression in the tongue squamous cell carcinoma cell line and tissue specimen was also detected by quantitative PCR.Results:Quantitative PCR showed that mitt-25-3p expression in the tongue squamous cell carcinoma cell lines and tissue specimen was significantly lower than that in the adjacent tissue.MTT and cell colony formation assays showed that after miR-25-3p overexpression,the proliferation of transfected Tca8113 was obviously attenuated.Western blot analysis and quantitative PCR showed that after miR-25-3p overexpression.p21^(cipt)and p27^(kipt)expressions were upregulated,while cyclinD1,AKT,FOXO1 expressions were downregulated,and AKT and FOXO1 phosphorylation was inactivated in the transfected Tca8113 cells.Conclusions:MiR-25-3p inhibited the proliferation of tongue squamous cell carcinoma cells and regulated cell cycle-related protein expression,playing an important role in the occurrence and development of squamous cell carcinoma of the tongue.

Studies on Breeding and Application of Thermo-sensitive Genie Male Sterile Line 402S in Brassica napus Ⅰ. Hie Breeding Procedure and Character of 402S

A new rapeseed TGMS line 402S has been bred through six years with seven generations by hybridization and systematic breeding.402S possesses the similar thermo-sensitive male sterility to its female parent Xiangyou 91S which performed fertile,partially sterile and sterile throughout the flowering stage,and resembles its male parent Zhongshuang 4 on agronomic traits.And 402S has the double-low (low erutic acid and low glucosinolate) quality like its original parents.

Effect of N-tosyl-L-phenylalanylchloromethyl Ketone on Tumor Necrosis Factor-alpha-induced NF-κB Activation and Apoptosis in U937 Cell Line

Summary: To investigate the effect of N-tosyl-L-phenylalanylchloromethyl ketone (TPCK) on tumor necrosis factor-alpha-induced NF-κB activation and apoptosis in U937 cell line, changes and subcellular localization of NF-κB/p65 and IκB-α were observed by fluorescencemicroscopy and expression and degradation of IκB-α by flow cytometry. The apoptosis of U937 cells was measured by flow cytometry and electrophoresis of DNA. Immunolfluorescence assay showed that NF-κB/p65, IκB-α only localized in cytoplasm. After TNF-α stimulation, p65 was localized only in nuclei, and IκB-α was only localized in cytoplasm and decreased. The changes of TNF-α stimulation were specifically inhibited by TPCK. Flow cytometry also revealed the downregulation of IκB-α protein during TNF-α-induced apoptosis and the down-regulation was specifically inhibited by TPCK. Flow cytometry also showed the apoptosis of U937 cells after TNF-α induction. DNA ladder can be detected in cells treated by TNF-α. It is concluded that degradation of IκB-α protein and NF-κB/p65 translocation occur during TNF-α-induced apoptosis of U937 cells, suggesting the activation of NF-κB. TPCK-sensitive protease plays an important role in the degradation of IκB-α protein induced by TNF-α in U937 cells. TPCK sensitive protease also plays an important role in the apoptosis of U937 cells induced by TNF-α.

The effect of adenovirus expressing wild-type p53 on 5-fluorouracil chemosensitivity is related to p53 status in pancreatic cancer cell lines

AIM:There are conflicting data about p53 function on cellular sensitivity to the cytotoxic action of 5-fluorouracil (5-FU). Therefore the objective of this study was to determine the combined effects of adenovirus-mediated wild-type (wt) p53 gene transfer and 5-FU chemotherapy on pancreatic cancer cells with different p53 gene status. METHODS:Human pancreatic cancer cell lines Capan-1^(p53mut), Capan-2^(p53wt),FAMPAC^(p53mut),PANC1^(p53mut),and rat pancreatic cancer cell lines AS^(p53wt) and DSL6A^(p53null) were used for in vitro studies.Following infection with different ratios of Ad- p53-particles (MOI) in combination with 5-FU,proliferation of tumor cells and apoptosis were quantified by cell proliferation assay (WST-1) and FACS (PI-staining).In addition,DSL6A syngeneic pancreatic tumor cells were inoculated subcutaneously in to Lewis rats for in vivo studies. Tumor size,apoptosis (TUNEL) and survival were determined. RESULTS:Ad-p53 gene transfer combined with 5-FU significantly inhibited tumor cell proliferation and substantially enhanced apoptosis in all four cell lines with an alteration in the p53 gene compared to those two cell lines containing wt-p53.In vivo experiments showed the most effective tumor regression in animals treated with Ad-p53 plus 5-FU.Both in vitro and in vivo analyses revealed that a sublethal dose of Ad-p53 augmented the apoptotic response induced by 5-FU. CONCLUSION:Our results suggest that Ad-p53 may synergistically enhance 5-FU-chemosensitivity most strikingly in pancreatic cancer cells lacking p53 function.These findings illustrate that the anticancer efficacy of this combination treatment is dependent on the p53 gene status of the target tumor cells.

Stark-Broadened Profiles of the Spectral Line P_α in He Ⅱ Ions

We report a systematic method to perform calculations of spectral line broadening parameters in plasmas. This method is applied to calculate Stark-broadening line profiles of Pα（n = 4 → n = 3） transitions under certain specific plasma conditions, by treating this case as an example. In the framework of the fully relativistic Dirac R- matrix theory, we calculate the electron-impact broadening operators, which are assumed to be diagonal matrix to simplify the situation. The electric microfield distribution function is calculated by retaining Hooper＇s formalism. The dipole matrix elements and atomic structure parameters used in these calculations have been obtained from atomic structure GRASP code. Based on this required data, we calculate the Stark-broadened line profiles of the Paschen spectral lines in He Ⅱ ions in a systematic manner. Overall, there is a very good agreement between our calculated Stark-broadened line profiles and other line Our reported spectral line-broadening data have real also play a fundamental role in plasma modeling. broadening numerical simulation codes （Sire U and MELS）. applications in plasma spectroscopy, plasma diagnosis and

Role of stress-activated MAP kinase P38 in cisplatin-and DTT-induced apoptosis of the esophageal carcinoma cell line Eca109

AIM： To study the role of P38 kinase in esophageal cancer cell apoptosis induced by genotoxin, cisplatin and the unfolded protein response （UPR） inducer, dithiothreitol （DTT）. METHODS： Esophageal carcinoma cell line Eca109 was cultured in RPMI 1640 medium to 70% confluency and treated with either cisplatin, DTT, or cisplatin plus DTT in the presence or absence of P38 inhibitor, SB203580. The untreated cells served as the control. The esophageal carcinoma cell apoptosis was detected by agarose gel DNA ladder analysis and quantified by flow cytometry. The P38 phosphorylation was detected by immunohistochemistry using antibodies specific to phosphorylated P38 protein. RESULTS： （1） Both cisplatin and DTF induced apoptosis in the esophageal cancer cell line Eca109 as shown by DNA ladder formation; （2） As detected by antibodies specific for the phosphorylated P38 protein （p-P38）, both cisplatin and DTT treatments activated the stress-activated enzyme, MAP kinase P38. The number of positive cells was about 50% for the treatment groups, comparing to that of 10% for untreated group. DTF treatment, but not cisplatin treatment, induces nuclear localization of p-P38; （3） As measured by flow cytometry, inhibition of P38 activity by SB203580 blocks DTT- and cisplatin-induced apoptosis. The rates for DTT, cisplatin, and DTT plus cisplatin-induced apoptosis were 16.8%, 17.1%, and 21.4%, respectively. Addition of the SB compound during the incubation reduced the apoptotic rate to about 7.6% for all the treatment groups, suggesting that P38 activation is essential for cisplatin- and DTT-induced apoptosis in Eca109 cells. CONCLUSION： （1） Both DTT and cisplatin were able to induce apoptosis in esophageal cancer cell line Eca109; （2） P38 MAP kinase is essential for DTT- and cisplatininduced apoptosis in Eca109 cells; （3） P38 activation may be the common signaling component relaying the multiple upstream signaling events to the downstream cell death program.

Mutations and altered expression of p16^(INK4a) in human pancreatic carcinoma cell lines with different potential of metastasis

Objective:To analyze the p16INK4a genomic alteration and expression status in 3 human pancreatic carcinoma cell lines with different potential of metastasis. Methods:Using PCR-SSCP, Dot-blot and immunohistochemistry, the p16INK4a genomic mutation and expression were analyzed on DNA, mRNA and protein levels in 3 human pancreatic carcinoma cell lines Patu8902, Patu8988 and SW1990, which had different potential of metastasis. Results: (1) On DNA level: there was no deletion of p16INK4a Exon Ⅰ in 3cell lines; p16INK4a Exon Ⅱ was only deleted in Patu8902 while no deletion in Patu8988 and SW1990. No insertion, microdeletion and point mutation were found in the 3 cell lines. (2) On RNA level: the expression of p16INK4a protein was negative in Patu8902, low expressed in SW1990, but highly expressed in Patu8988.(3) On protein level: P16 protein was strongly stained in Patu8988, much lower in SW1990, but not stained in Patu8902. Conclusion:The genomic type and expression of p16INK4a are quite different in 3 pancreatic carcinoma cell lines which have different potential of metastasis. It is suggested that genomic homozygous deletion and low expression of mRNA might relate to the potential of metastasis of pancreatic cell lines. In other words, dysfunction of p16INK4a might be an important mechanism in the metastasis of pancreatic carcinoma.

Effects of Terminalia arjuna bark extract on apoptosis of human hepatoma cell line HepG2

AIM：To investigate the effects of Terminalia arjuna （T. arjuna） extract on human hepatoma cell line （HepG2） and its possible role in induction of apoptosis.METHODS： Human hepatoma cells were treated with different concentrations of ethanolic extract of T. arjuna and its cytotoxicity effect was measured by trypan blue exclusion method and lactate dehydrogenase leakage assay. Apoptosis was analyzed by light and fluorescence microscopic methods, and DNA fragmentation. The mechanism of apoptosis was studied with expression of p53 and caspase-3 proteins. Glutathione （GSH） content was also measured in HepG2 cells after T. arjuna treatment.RESULTS： T. arjuna inhibited the proliferation of HepG2 cells in a concentration-dependent manner. Apoptotic morphology was observed in HepG2 cells treated with T. arjuna at the concentrations of 60 and 100 mg/L. DNA fragmentation, accumulation of p53 and cleavage of procaspase-3 protein were observed in HepG2 cells after the treatment with T. arjuna. The depletion of GSH was observed in HepG2 cells treated with T. arjuna.CONCLUSION： T. arjuna induced cytotoxicity in HepG2 cells in vitro. Apoptosis of HepG2 cells may be due to the DNA damage and expression of apoptotic proteins. Depletion of GSH may be involved in the induction of apoptosis of HepG2 cells.

RADIATION-INDUCED APOPTOSIS OF TWO NASOPHARANGEALCARCINOMA CELL LINES

Objective: To study apoptosis induced by radiation in two nasopharyngeal carcinoma (NPC) cell lines, CNE and CNE-2. Methods: Hoechst 33342 staining, immuno-histochemical staining, RT-PCR, DNA dot blotting and Southern blotting were used to identify apoptosis. Results: A single dose of X-irradiation resulted in apoptosis, the apoptotic index (AI) was time- and dose-dependent. Different apoptotic responses existed in the two cell lines. Immunohistochemical staining showed that bcl-2 protein was strongly positive in CNE but negative in CNE-2. However, RT-PCR revealed p53 mRNA in CNE-2 but not in CNE. P53 and bcl-2 genes were both present in the two cell lines as shown by DNA blotting, but the 2.8 kb fragment of the p53 gene was much lower than the 5.6 kb fragment on CNE which was clearly shown in Southern hybridization, suggestive of partial deletion of p53 gene in CNE. Conclusion: Apoptotic response to radiation is different in two NPC cell lines. CNE is more radioresistant than CNE-2. Overexpression of bcl-2 protein and partial deletion of p53 gene may explain their difference in radiosensitivity.

c-Ha-ras and c-myc antisense oligodeoxynucleotides inhibit the proliferation and DNA synthesis in human gastric carcinoma cell lines

The effects of two antisense oligodeoxynucleotides on the expression of c-Ha-ras proto-oncogene and the growth of human gastric carcinoma cell lines were observed. Synthetic 15-mer directed at the region of the translational initiation site of c-Ha-ras proto-oncogene (ASO-r) greatly inhibited the proliferation (55. 61%,P<0. 05) and DNA synthesis (76. 79%,P<0. 05) of MGc-803 cell line. It also inhibited the proliferation (62. 02%,P<0. 05) and DNA synthesis (76. 78%, P<0. 05) of SGc-7901 cell line. A reduction in intracellular P21 ras protein levels in MGc-803 cell line was observed 6 h after the treatment with ASO-r and maintained over 12 h. Another synthetic 15-mer targeted against the initiation codon and downstream 4 codons of c-myc proto-oncogene (ASOm) inhibited only DNA synthesis of MGc-803 cell line (71. 37%, P<0. 05). The control 15-mer did not inhibit the expression of P21 protein and proliferation of these cell lines. These experiments seemed to provide evidence that ASO-r could be effective in inhibiting the expression of c-Ha-ras proto-oncogene and controlling the growth of human gastric carcinoma cells,and that the over-expression of c-Ha-ras proto-oncogene might mainly be associated with the malignant proliferation of human gastric carcinoma cells.

Studies on Breeding and Application of Thermo-sensitive Genie Male Sterile Line 402S in Brassica napus Ⅱ.Evaluation of Heterosis and Combining Ability

Augmented randomized complete block test was conducted to evaluate the heterosis of 80 hybrid combinations from TGMS line 402S and its original parent Xiangyou 91S,and the combining a-bility of 40 testcrossing lines.The results of identification test showed that among 47 combinations yielding over the control Xiangyou 15,17 ones with 402S and 3 ones with Xiangyou 91S overyielded more than 20%,reaching the significant level of 1 %;and among 51 combinations yielding over their corresponding higher yield parents,18 ones with 402S and 9 ones with Xiangyou 91S overyielded at 5% or 1% significant level.The test for the GCA effect of all parents indicated that 402S possessed a stronger combining ability than Xiangyou 91S on yield,ailiquae of main inflorescence,total siliquae per plant,seed yield of single plant and 1000 seed weight.10 testcrossing lines with high GCA were picked out for next testcrosses.Among 8 agronomic traits,total siliquae per plant and seed yield of single plant were regarded as the key selecting indexes according to the correlation analysis between yield and the agronomic traits on heterosis and on the GCA effect of all parents.

Effects of 4-(3-Chloro-Benzyl)-6,7-Dimethoxy-Quinazoline on Kinetics of P120-Catenin and Periplakin in Human Buccal Mucosa Squamous Carcinoma Cell Line

In order to detect molecular markers for the epidermal growth factor inhibitor 4-(3-chloro-benzyl)- 6,7-dimethoxy-quinazoline (tyrphostin), we investigated the kinetics of p120-catenin and periplakin in the human buccal mucosa squamous cancer cell line BICR 10 treated with 3 nM tyrphostin. Growth of BICR 10 cells was inhibited by treatment with tyrphostin. Although changes were not observed in the expression of EGFR and p120-catenin, expression of Akt, Src and periplakin in BICR 10 treated with 3 nM tyrphostin tended to decrease. In addition, phosphorylation of EGFR, Akt and Src was inhibited by treatment with tyrphostin. On immunocytochemical staining, immunoreactions with phosphorylated EGFR, phosphorylated Akt and phosphorylated p120-catenin were weak in BICR 10 treated with tyrphostin. There was a slight immunocy to chemical reaction to periplakin in BICR 10 cells induced by tyrphostin. In conclusion, the decrease in phosphorylation in EGFR and p120-catenin by tyrphostin, following the decrease in Src or Akt phosphorylation, may inhibit expression of several growth factors associated with the proliferation and migration of cancer cells.