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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Effects of Added Vegetable Oils on In vitro Formation of Fatty Acid Soaps and Fermentation Characteristics and NDF Disappearance Rate (식물성유 첨가가 In vitro 발효성상, NDF 소실율 및 지방산염 형성에 미치는 영향)

  • Kim, D.I.;Choi, J.R.;Lee, Y.H.;Lee, J.K.;Chung, T.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.355-372
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    • 2004
  • In vitro experiments were conducted to determine the formation of fatty acid soaps (FAS) and neutral detergent fiber (NDF) disappearance rate. The substrates were a basal alfalfa hay containing 1) no oil, 2) 10% soybean oil, 3) 10% com oil, on a weight basis. All the substrates were incubated in triplicate for 0, 3, 6, 12, 24 and 48h in each experiment. After the incubation in the first experiment serum bottles (6oml) were analyzed for nonesterified, esterifed and fatty acid soaps contents. The serum bottles (120mI) from the second experiment were analyzed for pH, $NH_3-N$ and VFA concentration, and dry matter and NDF disappearance rate. pH decreased and the concentration of NH3-N increased significantly with longer incubation time (P<0.0001). The disappearance rates of dry matter and NDF significantly varied with feed, incubation time and oils (P<0.05). The molar concentration of total VFA increased and proportion of acetate significantly decreased with incubation time (P<0.0001), but the proportion of propionate significantly increased with longer incubation time (P<0.0001). Addition of oils to diet lowered the ratio of acetate:propionate (P<0.05). The esterified fatty acids (EFA) decreased with increasing incubation time (P<0.0001), and nonesterified fatty acids (NEFA) increased due to lipolysis of EFA, NEFA then reacted with cations to increase formation of FAS. The formation of FAS increased significantly at 48h of incubation time (P<0.0001). Especially, formation of stearic acid soaps was 27.5 and 32.5 folds with soybean oil and com oil supplements, respectively, by 48h of incubation time (P<0.0001). Alfalfa hay had higher cation contents, particularly Ca, which react with NEFA and FAS can be formed with longer incubation time. Saturated fatty acids had a higher proportion of FAS than did unsaturated fatty acids, suggesting that the former may react more extensively with cations. FAS contents increased with increasing chain length of the fatty acids. Since added vegetable oils fonned FAS, it might decrease negative effects on in vitro fermentation characteristics and NDF disappearance rate.

Cloning, cSNP Identification, and Genotyping of Pig Complement Factor B(CFB) Gene Located on the SLA Class III Region (SLA Class III 영역의 돼지 Complement Factor B(CFB) 유전자의 Cloning, cSNP 동정 및 유전자형 분석)

  • Kim, Jae-Hwan;Lim, Hyun-Tae;Seo, Bo-Yeong;Zhong, Tao;Yoo, Chae-Kyoung;Jung, Eun-Ji;Jeon, Jin-Tae
    • Journal of Animal Science and Technology
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    • v.50 no.6
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    • pp.753-762
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    • 2008
  • The primers for RT-PCR and RACE-PCR were designed by aligning the pig genomic sequence and the human complement factor B(CFB) coding sequence(CDS) from the GenBank. Each PCR product was amplified in pig cDNA and sequencing was carried out. The CDS length of pig CFB gene was determined to be 2298 bp. In addition, the pig CDS was more longer than human and mouse orthologs because of insertion and deletion. The identities of porcine nucleotide sequences with those of human and mice were 84% and 80%, and the identities of amino acids were 79% to 77%, respectively. Three complement control protein(CCP) domains, one Von Willebrand factor A(VWFA) domain and a serine protease domain, that are revealed typically in mammals, were found in the pig CFB gene. Based on the CDSs determined, the primers were designed in intron regions for amplification of entire length of exons. In amplification and direct sequencing with genomic DNAs of six pig breeds, three cSNPs(coding single nucleotide polymorphisms) were identified and verified as missense mutations. Using the Multiplex-ARMS method, we genotyped and verified the mutations identified from direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS with two randomly selected DNA samples. The genotype of each sample exhibited the same results using both methods. Therefore, three cSNPs were identified from pig CFB gene and that can be used for haplotype analysis of the swine leukocyte antigen(SLA) class III region. Moreover, the results indicate that the Multiplex-ARMS method should be powerful for genotyping of genes in the SLA region.

Analysis of Vasopressin Receptor Type 2(AVPR2) Gene in a Pedigree with Congenital Nehrogenic Diabetes Insipidus : Identification of a Family with R202C Mutation in AVPR2 Gene (신성요붕증 가계에서 바소프레신 V2 수용체(AVPR2) 유전자 분석 : AVPR2 유전자 R202C 돌연변이의 발견)

  • Park June-Dong;Kim Ho-Sung;Kim Hee-Joo;Lee Yoon-Kyung;Kwak Young-Ho;Ha Il-Soo;Cheong Hae-Il;Choi Yong;Park Hye-Won
    • Childhood Kidney Diseases
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    • v.3 no.2
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    • pp.209-216
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    • 1999
  • Purpose : Nephrogenic diabetes insipidus (NDI) is a rare X-linked disorder associated with renal tubule resistance to arginine vasopressin (AVP). The hypothesis that the defect underlying NDI might be a dysfunctional renal AVPR2 has recently been proven by the identification of mutations in the AVPR2 gene in NDT patients. To investigate the association of mutations in th AVPR2 gene with NDI, we analyzed the AVPR2 gene located on the X chromosome. Methods : We have analyzed the AVPR2 gene in a kindred with X-linked NDI. The proband and proband's mother were analyzed by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP) and DNA sequencing of the AVPR2 gene. We also have used restriction enzyme analysis of genomic PCR product to evaluate the AVPR2 gene. Results : C to T transition at codon 202, predictive of an exchange of tryptophan 202 by cysteine(R202C) in the third extracellular domain was identified. This mutation causes a loss of Hae III site within the gene. Conclusion : We found a R202C missense mutation in the AVPR2 gene causing X-linked NDI, and now direct mutational analysis is available for carrier screening and early diagnosis.

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Production of hTPO Transgenic Chickens using Tetracycline-Inducible Expression System (Tetracycline-Inducible Expression System을 이용한 Human Thrombopoietin (hTPO) 형질전환 닭의 생산)

  • Kwon, M.S.;Koo, B.C.;Kim, D.H.;Kim, M.J.;Kim, T.
    • Korean Journal of Poultry Science
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    • v.36 no.2
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    • pp.177-186
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    • 2009
  • It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

Experimental Pathogenesis of Pullorum Disease with the Local Isolate of Salmonella enterica serovar. enterica subspecies Pullorum in Pullets in Bangladesh

  • Haider, M.G.;Chowdhury, E.H.;Khan, M.A.H.N.A.;Hossain, M.T.;Rahman, M.S.;Song, H.J.;Hossain, M.M.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.341-350
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    • 2009
  • The research work was carried out to study the pathogenesis covering the clinical signs, gross and histopathological lesions in different organs, and reisolation and identification of the organisms after experimental infection with the local isolate of Salmonella enterica serovar. enterica subspecies (S.) Pullorum at different time interval of the experiment during the period February 2006 to December 2006. One hundred pullets (seronegative to S. Pullorum of 12 weeks age were purchased and divided into 5 (A, B, C, D and E) groups and each group consisted of 20 birds. Four groups (A, B, C and D) were infected orally with a dose of $10^6\;CFU$, $10^7\;CFU$, $2{\times}10^7\;CFU$, $10^8\;CFU$ of S. Pullorum, respectively, and one group (E) was treated as uninfected control. The used methods were necropsy and histopathology, culture of bacteria, staining and biochemical test of Salmonella. Five birds from each group were randomly selected and sacrificed $1^{st}$ week, $2^{nd}$, $3^{rd}$ and $4^{th}$ weeks of post infection (PI). From all the groups, the bacteriological samples (crop, liver, lung, heart, spleen, bile duodenum, ceca and blood) were collected with pre enriched in buffered peptone water in sterile poly bags. Liver, lungs, heart, spleen, intestine, etc. were collected in 10% buffered-formalin for histopathological examination. No clinical signs, gross and histopathological lesions were found in control group and no S. Pullorum was reisolated. Clinical sign of experimentally infected with S. Pullorum in pullets were loss of appetite (100%), slight depression (75%), ruffled feathers (85%), diarrhea (60%) and loss of weight (100%) in chickens. The feed intake and body weight at different weeks after PI differed significantly (p<0.01) among the groups. Grossly, the highest recorded lesion was button-like ulcer in the ceca (80%) and the lowest was white nodules in lungs (1.25%). S. Pullorum were reisolated from crop (91.25%), liver (91.25%), lung (83.75%), heart (71.25%), spleen (87.75%), bile (33.25%), duodenum (92.50%), ceca (97.50%) and from different group of infection (61.25%). The highest microscopic findings were intestinal and cecal mucosa and submucosa exhibited infiltration of mononuclear cells and congestion (96.25%), and the lowest finding was nodule formation in the lungs (3.75%). The pattern of the disease production by local isolate of S. Pullorum in Bangladesh is almost similar with other isolates in different countries.

Effect of the Induced Molting with Wheat Bran on the Postmolt Performance of Laying Hens (밀기울 위주의 환우용 사료가 산란계의 환우 후 생산성에 미치는 영향)

  • HwangBo, Jong;Hong, Eui-Chul;Kang, Bo-Seok;Kim, Hak-Kyu;Heo, Kang-Nyeong;Choo, Hyo-Jun;Na, Jae-Cheon;Choi, Yang-Ho;Kim, Won
    • Korean Journal of Poultry Science
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    • v.38 no.3
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    • pp.205-211
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    • 2011
  • This work was carried out to investigate the effect of the induced molting diet based on wheat bran on the postmolt performance of layers. Two hundred White Leghorn layers (65-old-wk) with over 80% egg production were used for 8 weeks in this work. Treatments were non-molt control (CO), fasting treatment for 10 days (FW), molt treatment with used molting diet for 4 wk (UM), molt treatment with molting diet based on corn-wheat bran for 4 wk (CW), and molt treatment with molting diet based on wheat bran for 4 wk (WM) as 5 treatments (4 replications/treatment and 10 birds/replication). Feed intake decreased at molting treatments at first weeks and increased after the 3rd week compared to control (P<0.05). Body weight (BW) loss were 18.6% of initial BW at first week in FW treatment, and were 11.4, 14.2 and 17.4% in UM, CW and WM treatments at 4th weeks (P<0.05). Egg production decreased at 1 week in molt treatment and stopped at 2 week in FW, whereas, other molting treatments didn't stop laying eggs. The birds started to lay egg at 4 week in FW and recovered at 5 week in other molting treatments. Egg quality (eggshell thickness, eggshell breaking strength, haugh unit) was high in molting treatments compared with control at 8th weeks (P<0.05). Finally, molting diet based on wheat bran affected BW loss and egg quality such as eggshell thickness, eggshell breaking strength and haugh unit.

Design and Implementation of Game Server using the Efficient Load Balancing Technology based on CPU Utilization (게임서버의 CPU 사용율 기반 효율적인 부하균등화 기술의 설계 및 구현)

  • Myung, Won-Shig;Han, Jun-Tak
    • Journal of Korea Game Society
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    • v.4 no.4
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    • pp.11-18
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    • 2004
  • The on-line games in the past were played by only two persons exchanging data based on one-to-one connections, whereas recent ones (e.g. MMORPG: Massively Multi-player Online Role-playings Game) enable tens of thousands of people to be connected simultaneously. Specifically, Korea has established an excellent network infrastructure that can't be found anywhere in the world. Almost every household has a high-speed Internet access. What made this possible was, in part, high density of population that has accelerated the formation of good Internet infrastructure. However, this rapid increase in the use of on-line games may lead to surging traffics exceeding the limited Internet communication capacity so that the connection to the games is unstable or the server fails. expanding the servers though this measure is very costly could solve this problem. To deal with this problem, the present study proposes the load distribution technology that connects in the form of local clustering the game servers divided by their contents used in each on-line game reduces the loads of specific servers using the load balancer, and enhances performance of sewer for their efficient operation. In this paper, a cluster system is proposed where each Game server in the system has different contents service and loads are distributed efficiently using the game server resource information such as CPU utilization. Game sewers having different contents are mutually connected and managed with a network file system to maintain information consistency required to support resource information updates, deletions, and additions. Simulation studies show that our method performs better than other traditional methods. In terms of response time, our method shows shorter latency than RR (Round Robin) and LC (Least Connection) by about 12%, 10% respectively.

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A Clinical Study for Promoting Quality Nusing Care in a University Hospital (질적 간호제공을 위한 간호단위 시범 운영 효과에 관한 임상적 연구)

  • Lee, A.J.;Kim, S.H.;Seong, Y.H.;Yoo, S.A.;Kwon, I.G.;Jeong, Y.I.;Nam, H.K.;Kwon, E.J.
    • The Korean Nurse
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    • v.32 no.5
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    • pp.66-77
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    • 1994
  • The purpose of this study was to develop a new nursing unit which can meet changing health care needs, enhance patients' satisfaction and nurses' job satisfaction, and finally guarantee quality nursing care with present manpower. For this, one medical unit was selected as a unit for quality care. And one medical unit which is similar in staffing and patients' characteristics was selected as a control unit. To assess present problems and identify the remedies to the problems a hospital-wide survey and a workshop were performed. According to the survey results, educational programs and improvement of the facilities and equipment supply system, managereal support for interdepartmental cooperation and intensification of bed-side nursing care were adopted as main principles for operating model unit, This model unit was operated for 3 months from Sep. 1, 1992 to Nov. 30, 1992. To evaluate the effectiveness of the model unit, derect/indirect nursing care hours, patients' satisfaction to nursing care, nurses' job satisfaction, and quality care index were measured. Direct/indirect nursing care hours were compared with that of the control unit, and patients' and nurses' satisfaction and quality care index were measured before and after operating model unit and compared with each other. The results of the study were as follows; 1. In the model unit mean direct nursing care hours per cach shift was 146.88 minutes and indirect nursing care hours was 354.72 minutes. The ratio of the direct nursing care hour to indirect nursing hour was 29.6 ; 70.4 and that of the control unit was 26.9 : 73.1. Direct nursing care hour in model unit was longer than that of the control unit. But, the difference was not significant. In subcategories of direct nursing care, the time spent in mobility and exercise, conservation of body temperature, hygiene, and communication and health education were longer than that of the con" trol unit. 2. Indirect nursing care hour in model unit was shorter than that of the control unit. But, the difference was not significant. In subcategories of indirect nursing care, the time spent in drug management and ward arrangement was shorter than that of the control unit. 3. Patients' satisfaction to nursing care was increased significantly after operating the model unit (T=-3.48, P=-0.002) and satisfaction to subcategories of physical comfort measure, psychological cate, and unit management components were significantly higher than before. 4. In the model unit, nurses' total job satisfaction was increased significantly after operating the model unit(Z=2.1004, P=.0357) and satisfaction to subcategory of satisfaction to administration was significantly higher than before (Z=-2.0732, P=.0382). 5. After operating the model unit, quality care index was increased from 89 to 93. With this results, it can be summarized that all the measures tried for quality care, such as educational programs, managereal support for interdepartmental cooperation, and improvement of the equipment and facility provision resulted in partial increase in direct nursing care hours, nurses satisfaction to their job and patients' satisfaction to nursing care. In can be postulated that managereal support and motivation without proper staff supplementation is not enough for increasing direct nursing care hours. And for the enhancement of the level in clinical nursing, and staff supplement must be considered sincerely and the measures for reducing indirect nursing care hours, such as computerization of nursing care activities, improvement of facilities and equipment and facilities supply system, must be instituted in addition.

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Effect of Feeding Rare Earth on Egg Production and Hatchability Broiler Growth (희토(稀土)의 급여가 종계의 산란율 및 부화율에 미치는 영향)

  • Ham, S.K.;Song, T.H.;Zhang, G.Q.;Hur, S.N.;Park, H.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.225-231
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    • 2006
  • To investigate the effect of feeding rare earth (RE) on the performance of breeder hens a feeding trial with sixteen thousands of 158 day old Ross broiler breeder hens was conducted for thirty weeks. A mixture of RE- chlorides containing mainly La, Ce, and Pr was mixed into corn-soy based diet at two levels, 300 and 600 mg/kg, All the hens were housed in flat layer houses and the ratio of male to female was maintained at one to ten. Dietary supplementation of the of RE at a level of 300 mg/kg made the hens reach egg production peak higher by about 6% and earlier by about two weeks. As a result, it made higher hatchable egg production by 3.5%. It also reduced dramatically the mortality of both male and female breeders(P<0.05). Egg weight was slightly increased but egg qualify was not much influenced by dietary supplementation of RE. Egg albumin hight and Haugh unit were significantly improved while egg shell thickness, egg breaking strength and yolk color were similar to those of the control. Experimental results appeared to show a good possibility that egg fertility and hatchability were improved by feeding RE. Dietary supplementation of RE at a level of 300 mg/kg should be acceptable but not the 600 mg/kg level for breeder hens. Further studies on the effect of RE on egg fertility and hatchability appears to be necessary.