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Articles by Lu Chen
Total Records ( 5 ) for Lu Chen
  Lu Chen , Kangle Yi , Yongfeng Sun , Yanling Sun , Lina Tang and Xu Zhou
  Neurotrophins (NTs) belong to a family of soluble homodimeric polypeptide growth factors and are widely recognized for their essential roles in central and peripheral nervous systems. One such neurotrophin, Brain-Derived Neurotrophic Factor (BDNF) was originally described in the nervous system but has now been shown to be expressed in reproductive system. In this study, the researchers examined the presence and different expression levels of BDNF mRNA in porcine reproductive organs during different stages of estrous cycle and in pig oocytes in Germinal Vesicle (GV) and in vitro matured Metaphase II (MII) stage. In oviduct and uterus, BDNF mRNA expression was higher than that in ovary (p<0.05). The expression level in luteal phase ovary is higher than that in follicular phase ovary but the difference between them was not significant (p>0.05), a similar but more significant change occurred in oviduct (p<0.05). However, the expression levels in uterus were on an opposite trend i.e., a higher level of mRNA for BDNF was found in follicular phase uterus instead of in luteal phase uterus. BDNF mRNA was also detected in GV oocytes and in vitro matured MII oocytes with significantly higher amounts in GV oocytes than in MII oocytes (p<0.01). These results suggest a possible role for BDNF in the regulation and modulation of pig reproductive function and oocyte maturation.
  Kangle Yi , Chunjin Li , Qili Qin , Lu Chen , Yanling Sun , Huahai Chen , Haifeng Yan , Deshun Shi and Xu Zhou
  The factor in the germline alpha (FIGLA) is a basic helix-loop-helix transcription factor that is essential for folliculogenesis and regulates expression of zona pellucida genes in mouse and human. But bovine homologue has not as yet been confirmed experimentally. Here, researchers present the first cDNA cloning and transcript expression analysis of the bovine FIGLA gene. Using RT-PCR and quantitative real-time PCR, researchers revealed that expression within adult cattle tissues is limited to the ovary. The researchers found that Germinal Vesicle (GV) oocytes, Metaphase II (MII) oocytes, 4 and 8 cell embryos, morula and blastocysts were all shown to express mRNA for FIGLA and expression were different in bovine oocytes and IVF embryos at different stages (p<0.05) with the highest expression in Germinal Vesicle (GV) oocytes and lower expression in 8 cells and blastocysts embryos. There was no difference in mRNA levels for FIGLA gene among bovine IVF, PA (Parthenogenetic Activated) and NT (Nuclear Transfer) blastocysts (p>0.05). In situ hybridization the FIGLA RNA was only localized in ovarian follicle. The persistence of FIGLA in adult cowes suggests that it may regulate additional pathways that are essential for bovine ovarian and embryonic development.
  Lina Tang , Lu Chen , Chunjin Li , Yongfeng Sun , Yanling Sun and Xu Zhou
  The development of reproductive system in prepubertal gilts was affected not only by gonadotropins but also by other cytokines and growth factors. Recent studies have revealed that Neurotrophins (NTs) which are well known for their essential roles in neural system also expressed in mammalian reproductive organs and involved in the regulation of mammalian reproduction. Dietary energy level is an important factor affecting domestic animal reproductive performance including central effects on Hypothalamic Pituitary Gonadal (HPG) axis and local effects on gonads. In the present study, the expression of Neurotrophin-4 (NT-4) which is a member of NTs in reproductive tissues of prepubertal gilts including ovary, oviduct and uterus was studied by immunohistochemistry. The effects of dietary energy on NT-4 mRNA expression were also investigated by quantitative PCR. The results show that NT-4 was localized in all classes of follicles in ovaries of prepubertal gilts including primordial follicles, primary follicles and secondary follicles, NT-4 was localized in mucosal epithelial cells in oviducts and glandular epithelium and endometrial epithelium in uteri. Furthermore, different dietary energy levels altered mRNA expression level of NT-4 in ovaries, uteri and oviducts of prepubertal gilts. NT-4 mRNA expression level in ovaries and uteri was higher in high dietary energy group than that in mid and low-energy groups. In oviduct, the expression pattern of NT-4 mRNA among three different dietary energy groups is similar as that occurred in in ovaries and uteri but with more significant differences. These results suggest that NT-4 may be one of the signals that links metabolic status and neuroendocrine control of reproduction in pig.
  Yongfeng Sun , Tongao Yang , Jingtao Hu , Zhe Hao , Yujian Sui , Yingying Fu , Lu Chen , Haiyang Zhu and Wei Wu
  The muscular tissue of breast was dissected from 8 weeks old Jilin White goose in the present study. The big fragment PCR Method was used to amplify double-strand cDNA based on the SMART techniques for construction of a full-length cDNA library. After digestion with restriction endonuclease Sfi |, a modified vector of pBluescript II SK-plasmid with the adaptors containing Sfi |A and Sfi |B sites was used to recombine with the cDNA products amplified. The recombinants were cloned by transformation into competent Escherichia coli DH2α. A plasmid cDNA library with goose muscle was constructed. The results showed that the titer of the cDNA library was 1.01x106 pfu mL-1 and the percentage of recombinant clones was 97%. The length of most cDNA inserted was between 0.25 and 1.6 kb identified by gel electrophoresis after cDNA PCR amplification. The unigene ratio was 66.7% and the percentage of complete cDNA sequences was 80% by estimating from the 24 clones sequenced randomly. It is helpful to study muscle development of goose at molecular level in the future.
  Taro Masuda-Sasa , Piotr Polaczek , Xiao P. Peng , Lu Chen and Judith L. Campbell
  The polyguanine-rich DNA sequences commonly found at telomeres and in rDNA arrays have been shown to assemble into structures known as G quadruplexes, or G4 DNA, stabilized by base-stacked G quartets, an arrangement of four hydrogen-bonded guanines. G4 DNA structures are resistant to the many helicases and nucleases that process intermediates arising in the course of DNA replication and repair. The lagging strand DNA replication protein, Dna2, has demonstrated a unique localization to telomeres and a role in de novo telomere biogenesis, prompting us to study the activities of Dna2 on G4 DNA-containing substrates. We find that yeast Dna2 binds with 25-fold higher affinity to G4 DNA formed from yeast telomere repeats than to single-stranded DNA of the same sequence. Human Dna2 also binds G4 DNAs. The helicase activities of both yeast and human Dna2 are effective in unwinding G4 DNAs. On the other hand, the nuclease activities of both yeast and human Dna2 are attenuated by the formation of G4 DNA, with the extent of inhibition depending on the topology of the G4 structure. This inhibition can be overcome by replication protein A. Replication protein A is known to stimulate the 5`- to 3`-nuclease activity of Dna2; however, we go on to show that this same protein inhibits the 3`- to 5`-exo/endonuclease activity of Dna2. These observations are discussed in terms of possible roles for Dna2 in resolving G4 secondary structures that arise during Okazaki fragment processing and telomere lengthening.
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