Intraspecific diversity of molluscan species is usually studied based on maternally inherited mitochondrial DNA,from which only part of the evolutionary history can be reflected.Some nuclear ribosomal RNA genes such a...Intraspecific diversity of molluscan species is usually studied based on maternally inherited mitochondrial DNA,from which only part of the evolutionary history can be reflected.Some nuclear ribosomal RNA genes such as 28S rRNA represent poten-tial candidates that can be easily applied in phylogeography because of lacking intraindividual variation.However,considering their low polymorphism,genetic appraisals on whether and how they can be used in population studies are necessary.Here,we applied a short 28S rRNA to assess genetic patterns of the clam Cyclina sinensis along the coast of China and compared the results with a for-mer study based on COI and ITS-1 analyses.The results revealed the 28S rRNA data set was characterized by an extremely low level of variation,with only seven haplotypes defined for 93 individuals.Haplotype and nucleotide diversity for each population was al-most the lowest when compared with the other two markers.However,the distribution of two dominant haplotypes showed clear geo-graphic patterns,and significant population differentiation was revealed between the East China Sea and the South China Sea.These patterns were highly concordant with findings of the former study that populations of C.sinensis were historically separated by land bridges among sea basins.Our study suggested that although the nuclear rRNAs have shortcomings such as low variation,they have advantages including lack of intraindividual variation and high amplification rates.Applying rRNA genes can enrich the toolbox of nuclear markers in molluscan phylogeographic studies.展开更多
AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: ...AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: NF-κB activation was assayed by immunohistochemical staining in formalin-fixed, paraffin-embedded specimens from 45 gastric carcinoma patients. Electrophoretic mobility shift assay (EMSA) method was used for nuclear protein from these fresh tissue specimens. Heparanase gene expression was quantified using quantitative RT-PCR.RESULTS: The nuclear translocation of RelA (marker of NF-κB activation) was significantly higher in tumor cells compared to adjacent and normal epithelial cells [(41.3±3.52)% vs (0.38±0.22) %, t= 10.993, P= 0.000<0.05; (41.3±3.52)%vs (0±0.31)%, t = 11.484, P = 0.000<0.05]. NF-κB activation was correlated with tumor invasion-related clinicopathological features such as lymphatic invasion,pathological stage, and depth of invasion (Z = 2.148,P= 0.032<0.05; χ2 = 8.758, P= 0.033<0.05; χ2 = 18.531,P = 0.006<0.05). NF-κB activation was significantly correlated with expression of heparanase gene (r= 0.194,P= 0.046<0.05).CONCLUSION: NF-κB RelA (p65) activation was related with increased heparanase gene expression and correlated with poor clinicopathological characteristics in gastric cancers. This suggests NF-κB as a major controller of the metastatic phenotype through its reciprocal regulation of some metastasis-related genes.展开更多
The nuclear capsid protein gene (vp39) or Bombyx mori nuclear polykedrosis virus (BmNPV)was amplified successfully by PCR technique and inserted into pGEM 3zf(+). The 5’ and 3’ terminal area orthe amplirled vp39 gen...The nuclear capsid protein gene (vp39) or Bombyx mori nuclear polykedrosis virus (BmNPV)was amplified successfully by PCR technique and inserted into pGEM 3zf(+). The 5’ and 3’ terminal area orthe amplirled vp39 gene were sequenced with sliver-staining dideoxy method. Bmvp39 gene was sub-cloned into the expression vector pRSET-A, and transformed into E. coli BL21. This gene was highly expressed byIPTG induction. SDS-PAGE analysis showed that the expressed protein is about 38 kd, and the expressedamount reached maxlum in 4 h with IPTG induction.展开更多
Objective To investgate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. Methods One hundred and ninety three CHD patients (94 males and 99 females) and their biol...Objective To investgate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. Methods One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method. Results In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+)in case fathers (5.0 %) was apparently lower than that in the control (9.1%, P=0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-)with OR 0.26 (95% CI: 0.11-0.60). Conclusion MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.展开更多
The nucleax mains attachment regions(MARs) and the binding nuclear matrix proteins in the 5’-flalildng cisacting elements of the humanε-globin gene have been examined. Using in vitro DNA-matrix binding assay,it has ...The nucleax mains attachment regions(MARs) and the binding nuclear matrix proteins in the 5’-flalildng cisacting elements of the humanε-globin gene have been examined. Using in vitro DNA-matrix binding assay,it has been shown that the positive stage-specific regulatory element (ε-PREII, -446bp-419bp) upstream of this gene could specifically associate with the nuclear matrix from K562 cells, indicating thatε-PREII mad be an erythroidspecilic facultstive MAR. In gel mobility shift assay and Southwestern blotting assal an eothroid-specific nuclear matrix protein (ε-NMPk) in K562 cells has been revealed to bind to this positive regulatory element (E-PREII). Furthermore, we demonstrated that the silencer (-392hp -177bp) uP8tream of the humanε-globin gene could associate with the nuclear matrices from K562, HEL and Raji cells. In addition, the nucleax matrix proteins prepared from these three cell lines could also bind to this silencer, suggesting that this silencer element linght be a constitutive nuclear mains attachment region (constitutive MAR). Our results demonstrated that the nucleax madrid and nuclear mains proteins lxilght play an important role in the regulation of the human 5-globin gene expression.展开更多
Harmful algal blooms (HABs) have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved an...Harmful algal blooms (HABs) have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene (pcna) was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates (89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively), and also 42%–43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages (r 2 =0.024 6). However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.展开更多
AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus ...AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus influenza and Escherichia coli infection was analyzed by employing quantitative Reverse Transcription Polymerase Chain Reaction techniques. Nucleofection was used to deliver Lenti-viral system to express or knock down Lpcat1 in MLE cells. Subcellular protein fractionation and Western blotting were utilized to study Lpcat1 nuclear relocation. RESULTS:Lpcat1 translocates into the nucleus from thecytoplasm in murine lung epithelia (MLE) after LPS treatment. Haemophilus influenza and Escherichia coli , two LPS-containing pathogens that cause pneumonia, triggered Lpcat1 nuclear translocation from the cytoplasm. The LPS inducible gene expression profile was determined by quantitative reverse transcription polymerase chain reaction after silencing Lpcat1 or overexpression of the enzyme in MLE cells. We detected that 17 out of a total 38 screened genes were upregulated, 14 genes were suppressed, and 7 genes remained unchanged in LPS treated cells in comparison to controls. Knockdown of Lpcat1 by shRNA dramatically changed the spectrum of the LPS inducible gene transcription, as 18 genes out of 38 genes were upregulated, of which 20 genes were suppressed or unchanged. Notably, in Lpcat1 overex-pressed cells, 25 genes out of 38 genes were reduced in the setting of LPS treatment.CONCLUSION:These observations suggest that Lpcat1 relocates into the nucleus in response to bacterial infection to differentially regulate gene transcriptional repression.展开更多
Homeobox A10(Hoxa10) gene is one of the most important candidate genes associated with the reproductive performance of humans and mice. Overexpression of Hoxa10 in mouse endometrium can increase litter size. Moreover,...Homeobox A10(Hoxa10) gene is one of the most important candidate genes associated with the reproductive performance of humans and mice. Overexpression of Hoxa10 in mouse endometrium can increase litter size. Moreover, Hoxa10 plays a key role in regulating the embryo implantation of sows. This study aimed to generate transgenic pigs using Hoxa10 via somatic cell nuclear transfer(SCNT). We established seven Hoxa10-transgenic cell lines, and two of the cell lines were selected as nuclear donors for the transfer. A total of 1 270 cloned embryos were generated and transferred to five surrogate mothers(Landrace×Yorkshire). Eight cloned male piglets were produced including one with cryptorchidism. Six transgenic piglets grew up healthy and produced 56 offspring. Finally, we obtained six transgenic male pigs and 26 transgenic positive offspring that can be used to further study the regulatory mechanism of Hoxa10 on the reproductive performance of pigs.展开更多
To explore a novel strategy for antisense gene therapy of cancer,the coding sequence of hum an proliferating cell nuclear antigen(PCNA) c DNA was reversely inserted into the eukaryotic vector p L XSN by molecular clon...To explore a novel strategy for antisense gene therapy of cancer,the coding sequence of hum an proliferating cell nuclear antigen(PCNA) c DNA was reversely inserted into the eukaryotic vector p L XSN by molecular cloning techniques and transferred into bladder cancer EJcells with li- posome. The PCNA expression in transferred cells was dynamically detected by immunofluo- rescence and RT- PCR techniques. Changes of proliferation activities of cancer cells were assayed by MTT colorim etric and cloning formation m ethods.In the experiment,the antisense eukaryotic vector was successfully constructed and nam ed as p L APSN.After transfection with it for1- 7 days,PCNA protein and m RNA levels in cancer cells were blocked by16 .74 % - 84 .2 1% (P< 0 .0 5 ) and2 3.2 7% - 86 .15 % (P<0 .0 5 ) respectively.The proliferation activities of transferred cells were inhibited by 2 7.91% - 6 2 .0 7% (P<0 .0 1) ,with cloning formation abilities being de- creased by 5 0 .81% (P<0 .0 1) . Itwas concluded that the in vitro proliferation activities of cancer cells could be effectively inhibited by blocking PCNA expression with antisense technique,which could serve as an ideal strategy for gene therapy of bladder cancer.展开更多
核输出蛋白是细胞核质运输中的重要蛋白。核输出蛋白1(export protein 1,XPO1)可介导包括p53在内的多种肿瘤抑制蛋白以及细胞周期调节蛋白的转运。抑制XPO1的转运功能可影响细胞增殖、分化、凋亡等过程,达到抗肿瘤目的。本文就核输出蛋...核输出蛋白是细胞核质运输中的重要蛋白。核输出蛋白1(export protein 1,XPO1)可介导包括p53在内的多种肿瘤抑制蛋白以及细胞周期调节蛋白的转运。抑制XPO1的转运功能可影响细胞增殖、分化、凋亡等过程,达到抗肿瘤目的。本文就核输出蛋白抑制剂在血液肿瘤常见基因突变和部分常用抗肿瘤药物耐药性方面的研究展开综述,探讨核输出蛋白抑制剂在血液肿瘤以及更多领域的应用。展开更多
Mitochondria are important in eukaryotic cells due to their functions in energy production and regulation over other cellular activities.Oocytes are produced by a long and precisely controlled process,the dysfunction ...Mitochondria are important in eukaryotic cells due to their functions in energy production and regulation over other cellular activities.Oocytes are produced by a long and precisely controlled process,the dysfunction of which leads to impaired female fertility.As oocytes mature,mitochondria are constantly under the regulation of nuclear genes,the process of which can be modulated by extracellular signals.Understanding how nuclear genes regulate mitochondrial functions is important for studying animal reproduction and human fertility.As more and more genes regulating mitochondrial functions in oocytes are being revealed,new approaches for improving female fertility in both human and animals through mitochondria can be developed.展开更多
Nucleotide sequences from 18S rDNA of 11 ostracodes, which represent four suborders and six superfamilies of podocopidan, were determined. The phylogenetic relationships were analyzed based on three kinds of methods (...Nucleotide sequences from 18S rDNA of 11 ostracodes, which represent four suborders and six superfamilies of podocopidan, were determined. The phylogenetic relationships were analyzed based on three kinds of methods (maximum-likelihood, maximum-parsimony, and neighbor-joining), and the three topologies gained were basically similar. The results have showed that (1) a monophyletic Podocopida was supported strongly; (2) the phylogenetic relationships of four suborders were (Darwinulocopina plus (Bairdiocopina plus (Cytherocopina plus Cypridocopina))), which indicated that a close relationship between Cytherocopina and Cypridocopina, and Darwinulocopina had separated early from the main podocopinan; (3) Cypridocopinan formed a monophyletic group, among which the phylogenetic relationship of three superfamilies was (Cypridoidea plus (Macrocypridoidea plus Pontocypridoidea)).展开更多
基金supported by research grants from the Science and Technology De-velopment Project of Weihai City(No.2018NS01)the In-dustrial Development Project of Qingdao City(No.20-3-4-16-nsh),and Guangxi Province(No.AA17204080-4).
文摘Intraspecific diversity of molluscan species is usually studied based on maternally inherited mitochondrial DNA,from which only part of the evolutionary history can be reflected.Some nuclear ribosomal RNA genes such as 28S rRNA represent poten-tial candidates that can be easily applied in phylogeography because of lacking intraindividual variation.However,considering their low polymorphism,genetic appraisals on whether and how they can be used in population studies are necessary.Here,we applied a short 28S rRNA to assess genetic patterns of the clam Cyclina sinensis along the coast of China and compared the results with a for-mer study based on COI and ITS-1 analyses.The results revealed the 28S rRNA data set was characterized by an extremely low level of variation,with only seven haplotypes defined for 93 individuals.Haplotype and nucleotide diversity for each population was al-most the lowest when compared with the other two markers.However,the distribution of two dominant haplotypes showed clear geo-graphic patterns,and significant population differentiation was revealed between the East China Sea and the South China Sea.These patterns were highly concordant with findings of the former study that populations of C.sinensis were historically separated by land bridges among sea basins.Our study suggested that although the nuclear rRNAs have shortcomings such as low variation,they have advantages including lack of intraindividual variation and high amplification rates.Applying rRNA genes can enrich the toolbox of nuclear markers in molluscan phylogeographic studies.
基金Supported by the Sate Key Basic Research and Development Plan of China (2003CB715904) and the National Science Foundation for 0verseas Distinguished Young Scholar (30428003)
文摘AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: NF-κB activation was assayed by immunohistochemical staining in formalin-fixed, paraffin-embedded specimens from 45 gastric carcinoma patients. Electrophoretic mobility shift assay (EMSA) method was used for nuclear protein from these fresh tissue specimens. Heparanase gene expression was quantified using quantitative RT-PCR.RESULTS: The nuclear translocation of RelA (marker of NF-κB activation) was significantly higher in tumor cells compared to adjacent and normal epithelial cells [(41.3±3.52)% vs (0.38±0.22) %, t= 10.993, P= 0.000<0.05; (41.3±3.52)%vs (0±0.31)%, t = 11.484, P = 0.000<0.05]. NF-κB activation was correlated with tumor invasion-related clinicopathological features such as lymphatic invasion,pathological stage, and depth of invasion (Z = 2.148,P= 0.032<0.05; χ2 = 8.758, P= 0.033<0.05; χ2 = 18.531,P = 0.006<0.05). NF-κB activation was significantly correlated with expression of heparanase gene (r= 0.194,P= 0.046<0.05).CONCLUSION: NF-κB RelA (p65) activation was related with increased heparanase gene expression and correlated with poor clinicopathological characteristics in gastric cancers. This suggests NF-κB as a major controller of the metastatic phenotype through its reciprocal regulation of some metastasis-related genes.
文摘The nuclear capsid protein gene (vp39) or Bombyx mori nuclear polykedrosis virus (BmNPV)was amplified successfully by PCR technique and inserted into pGEM 3zf(+). The 5’ and 3’ terminal area orthe amplirled vp39 gene were sequenced with sliver-staining dideoxy method. Bmvp39 gene was sub-cloned into the expression vector pRSET-A, and transformed into E. coli BL21. This gene was highly expressed byIPTG induction. SDS-PAGE analysis showed that the expressed protein is about 38 kd, and the expressedamount reached maxlum in 4 h with IPTG induction.
基金This work was supported by the Major State Basic Research Development Program of People's Republic of China (G1999055904) the Danone's Diet and Nutrition Research and Education Grant (DIC2002-08).
文摘Objective To investgate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. Methods One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method. Results In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+)in case fathers (5.0 %) was apparently lower than that in the control (9.1%, P=0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-)with OR 0.26 (95% CI: 0.11-0.60). Conclusion MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.
文摘The nucleax mains attachment regions(MARs) and the binding nuclear matrix proteins in the 5’-flalildng cisacting elements of the humanε-globin gene have been examined. Using in vitro DNA-matrix binding assay,it has been shown that the positive stage-specific regulatory element (ε-PREII, -446bp-419bp) upstream of this gene could specifically associate with the nuclear matrix from K562 cells, indicating thatε-PREII mad be an erythroidspecilic facultstive MAR. In gel mobility shift assay and Southwestern blotting assal an eothroid-specific nuclear matrix protein (ε-NMPk) in K562 cells has been revealed to bind to this positive regulatory element (E-PREII). Furthermore, we demonstrated that the silencer (-392hp -177bp) uP8tream of the humanε-globin gene could associate with the nuclear matrices from K562, HEL and Raji cells. In addition, the nucleax matrix proteins prepared from these three cell lines could also bind to this silencer, suggesting that this silencer element linght be a constitutive nuclear mains attachment region (constitutive MAR). Our results demonstrated that the nucleax madrid and nuclear mains proteins lxilght play an important role in the regulation of the human 5-globin gene expression.
基金The Nature Science Foundation of Qingdao, China under contract No. 05-1-JC-87International Foundation for Science under contract No.AA/16180 awarded to Sui Zhenghong
文摘Harmful algal blooms (HABs) have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene (pcna) was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates (89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively), and also 42%–43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages (r 2 =0.024 6). However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.
基金Supported by A United States National Institutes of Health R01 grant HL091916 to Zhao Yan American Heart Association grant 12SDG12040330 to Zou C, in part
文摘AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus influenza and Escherichia coli infection was analyzed by employing quantitative Reverse Transcription Polymerase Chain Reaction techniques. Nucleofection was used to deliver Lenti-viral system to express or knock down Lpcat1 in MLE cells. Subcellular protein fractionation and Western blotting were utilized to study Lpcat1 nuclear relocation. RESULTS:Lpcat1 translocates into the nucleus from thecytoplasm in murine lung epithelia (MLE) after LPS treatment. Haemophilus influenza and Escherichia coli , two LPS-containing pathogens that cause pneumonia, triggered Lpcat1 nuclear translocation from the cytoplasm. The LPS inducible gene expression profile was determined by quantitative reverse transcription polymerase chain reaction after silencing Lpcat1 or overexpression of the enzyme in MLE cells. We detected that 17 out of a total 38 screened genes were upregulated, 14 genes were suppressed, and 7 genes remained unchanged in LPS treated cells in comparison to controls. Knockdown of Lpcat1 by shRNA dramatically changed the spectrum of the LPS inducible gene transcription, as 18 genes out of 38 genes were upregulated, of which 20 genes were suppressed or unchanged. Notably, in Lpcat1 overex-pressed cells, 25 genes out of 38 genes were reduced in the setting of LPS treatment.CONCLUSION:These observations suggest that Lpcat1 relocates into the nucleus in response to bacterial infection to differentially regulate gene transcriptional repression.
基金supported by the National Major Special Project on New Varieties Cultivation for Transgenic Organisms,China(2014ZX08006-005 and 2014ZX0800950B)
文摘Homeobox A10(Hoxa10) gene is one of the most important candidate genes associated with the reproductive performance of humans and mice. Overexpression of Hoxa10 in mouse endometrium can increase litter size. Moreover, Hoxa10 plays a key role in regulating the embryo implantation of sows. This study aimed to generate transgenic pigs using Hoxa10 via somatic cell nuclear transfer(SCNT). We established seven Hoxa10-transgenic cell lines, and two of the cell lines were selected as nuclear donors for the transfer. A total of 1 270 cloned embryos were generated and transferred to five surrogate mothers(Landrace×Yorkshire). Eight cloned male piglets were produced including one with cryptorchidism. Six transgenic piglets grew up healthy and produced 56 offspring. Finally, we obtained six transgenic male pigs and 26 transgenic positive offspring that can be used to further study the regulatory mechanism of Hoxa10 on the reproductive performance of pigs.
基金This studywassupported by a grant from the NationalNatural Sciences Foundation ofChina(No. 39770 739)
文摘To explore a novel strategy for antisense gene therapy of cancer,the coding sequence of hum an proliferating cell nuclear antigen(PCNA) c DNA was reversely inserted into the eukaryotic vector p L XSN by molecular cloning techniques and transferred into bladder cancer EJcells with li- posome. The PCNA expression in transferred cells was dynamically detected by immunofluo- rescence and RT- PCR techniques. Changes of proliferation activities of cancer cells were assayed by MTT colorim etric and cloning formation m ethods.In the experiment,the antisense eukaryotic vector was successfully constructed and nam ed as p L APSN.After transfection with it for1- 7 days,PCNA protein and m RNA levels in cancer cells were blocked by16 .74 % - 84 .2 1% (P< 0 .0 5 ) and2 3.2 7% - 86 .15 % (P<0 .0 5 ) respectively.The proliferation activities of transferred cells were inhibited by 2 7.91% - 6 2 .0 7% (P<0 .0 1) ,with cloning formation abilities being de- creased by 5 0 .81% (P<0 .0 1) . Itwas concluded that the in vitro proliferation activities of cancer cells could be effectively inhibited by blocking PCNA expression with antisense technique,which could serve as an ideal strategy for gene therapy of bladder cancer.
文摘核输出蛋白是细胞核质运输中的重要蛋白。核输出蛋白1(export protein 1,XPO1)可介导包括p53在内的多种肿瘤抑制蛋白以及细胞周期调节蛋白的转运。抑制XPO1的转运功能可影响细胞增殖、分化、凋亡等过程,达到抗肿瘤目的。本文就核输出蛋白抑制剂在血液肿瘤常见基因突变和部分常用抗肿瘤药物耐药性方面的研究展开综述,探讨核输出蛋白抑制剂在血液肿瘤以及更多领域的应用。
基金the National Natural Science Foundation of China(Nos.41776144 and 32072954)Zhejiang Province Public Welfare Technology Application Research Project(including Natural Science Foundation)(No.LGF20C120001).
文摘Mitochondria are important in eukaryotic cells due to their functions in energy production and regulation over other cellular activities.Oocytes are produced by a long and precisely controlled process,the dysfunction of which leads to impaired female fertility.As oocytes mature,mitochondria are constantly under the regulation of nuclear genes,the process of which can be modulated by extracellular signals.Understanding how nuclear genes regulate mitochondrial functions is important for studying animal reproduction and human fertility.As more and more genes regulating mitochondrial functions in oocytes are being revealed,new approaches for improving female fertility in both human and animals through mitochondria can be developed.
基金supported by the Major Project of the National Natural Science Foundation of China under contract No.30130040the Cultivation Fund of the Key Scientific and Technical hmovation Project,Ministry of Education of China under contract No.704023+2 种基金E-institute of Shanghai Municipal Education Commission under contract No.E03009the Program of Shanghai Subject Chief Scientist under contract No.05XD14005 to Chen Liqiaothe PhD Program Scholarship Fund of ECNU 2005.
文摘Nucleotide sequences from 18S rDNA of 11 ostracodes, which represent four suborders and six superfamilies of podocopidan, were determined. The phylogenetic relationships were analyzed based on three kinds of methods (maximum-likelihood, maximum-parsimony, and neighbor-joining), and the three topologies gained were basically similar. The results have showed that (1) a monophyletic Podocopida was supported strongly; (2) the phylogenetic relationships of four suborders were (Darwinulocopina plus (Bairdiocopina plus (Cytherocopina plus Cypridocopina))), which indicated that a close relationship between Cytherocopina and Cypridocopina, and Darwinulocopina had separated early from the main podocopinan; (3) Cypridocopinan formed a monophyletic group, among which the phylogenetic relationship of three superfamilies was (Cypridoidea plus (Macrocypridoidea plus Pontocypridoidea)).