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Decreased Neutrophil Apoptosis in Patients with Sepsis is Related to the Activation of NF-κB (패혈증 환자에서 NF-κB 활성화에 의한 호중구 아포프토시스의 억제)

  • Kwon, Sung Youn;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.495-509
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    • 2003
  • Background : Neutrophil-mediated inflammation is usually self-limiting, because neutrophils have a remarkably short life span. Prolonged neutrophil survival, which is caused by decreased spontaneous apoptosis, leads to persistent inflammation in sepsis. Because many inflammatory cytokines, which generate signals that delay apoptosis, are regulated by nuclear factor-${\kappa}B$ transcription factor, we hypothesized that nuclear factor-${\kappa}B$ might be related to the reduced neutrophil apoptosis observed in sepsis. Methods : Neutrophils of healthy volunteers and sepsis patients were freshly isolated from venous blood. Neutrophil apoptosis was assayed with two approaches : by counting apoptotic cells under a microscope and by flow cytometry using Annexin V. The activity of nuclear factor-${\kappa}B$ was assessed by immunofluorescent staining or electrophoretic mobility shift assay. Expression of X-linked inhibitor of apoptosis was measured by western blot assay. Results : We confirmed reduced spontaneous neutrophil apoptosis in patients with sepsis. The number of apoptotic neutrophils in patients with sepsis increased to the level of that in healthy controls after cycloheximide treatment, suggesting that decreased spontaneous neutrophil apoptosis is dependent on de novo protein synthesis. In patients with sepsis, basal neutrophil nuclear factor-${\kappa}B$ was activated compared to the level in healthy controls. Moreover, a blockade of nuclear factor-${\kappa}B$ activity reversed the decreased spontaneous neutrophil apoptosis in sepsis patients. Meanwhile, X-linked inhibition of apoptosis expression, which is regulated by nuclear factor-${\kappa}B$, decreased 24 hours after incubation in healthy persons, but persisted for 24 hours in patients with sepsis. Conclusion : These observations suggest that the reduced spontaneous neutrophil apoptosis observed in patients with sepsis may be related to the induction of survival protein by nuclear factor-${\kappa}B$.

DNA Methylation of Multiple Genes in Gastric Cancer: Association with CpG Island Methylator Phenotype and Helicobocter pylori Infection (위암에서 유전자 메틸화와 CpG Island Methylator Phenotype 및 Helicobacter pylori균 감염과의 연관성)

  • Jun, Kyong-Hwa;Won, Yong-Sung;Shin, Eun-Young;Cho, Hyun-Min;Im, Myoung-Goo;Chin, Hyung-Min;Park, Woo-Bae
    • Journal of Gastric Cancer
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    • v.6 no.4
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    • pp.227-236
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    • 2006
  • Purpose: Methylation of gene regulatory elements plays an important role in gene inactivation without genetic alteration. Gastric cancer is one of the tumors that exhibit a high frequency of CpG island hypermethylation. The purpose of this study was to investigate the occurrence of CpG island hypermethylation in gastric carcinoma in relation to H. pylori infection, CIMP and clincopathologic variables. Materials and Methods: We investigated the promoter methylation Status of six genes (hMLH1, p16, p14, COX-2, MGMT, E-cadherin) and CIMP in 36 gastric carcinoma tissues as well as in nontumor tissues. CIMP status was investigated by examining the methylation status of MINT 1, 2, 12, 25 and 31. The methylation status of the promoter was examined by methylation-specific PCR (MSP) and H. pylori infection was examined by histological diagnosis after staining with Warthin-Starry silver. Results: Among the 36 gastric carcinoma tissues, DNA hypermethylation was detected in the following frequencies: 14 (38.9%) for p14, 13 (36.1%) for p16, 8 (22.2%) for MGMT, 10 (27.8%) for COX-2, 21 (58.3%) for E-cadherin, and 6 (16.7%) for hMLH1. The frequencies for MINT1 and MINT25 hypermethylation were significantly higher in tumor tissues than in nontumor tissues. 16 (44.4%) of the 36 gastric carcinoma tissues were positive for the CIMP CIMP-H tumors were associated with older patients and larger tumor size than CIMP-L tumors. We found a significant association between the presence of the CIMP and hypermethylation of p16. Hypermethylation of p16 and MINT2 were significantly different when compared by age. MINT1 gene methylation was significantly associated with H. pylori infection (P=0.004). Conclusion: Our results suggest that aberrant hypermethylation of multiple tumor related genes (hMLH1, p16, p14, COX-2, MGMT, E-cadherin, MINT1, 2, 12, 25, 31) occurs frequently in gastric carcinoma tissues. The hypermethylation of MINT1 was significantly higher in the tumor tissues and was associated with H. pylori infection.

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Expression of the Peroxisome-proliferator-activated $Receptor-{\gamma}$ in Human Gastric Cancer (위암에서 $PPAR-{\gamma}$ 발현의 임상적 의의)

  • Cho, Dong-Hui;Kang, Sung-Gu;Park, Sang-Su;Yoon, Jin;Kim, Il-Myung;Yoon, Seong-Min;Lee, Yun-Kyung;Lee, Yong-Jik;Yang, Dae-Hyun;Cho, Ik-Hang;Shin, Dong-Gue
    • Journal of Gastric Cancer
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    • v.6 no.4
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    • pp.250-256
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    • 2006
  • Purpose: Recently, interest in peroxisome-proliferator-activated receptors (PPAR) has increased, although clinical studies of the effect of $PPAR-{\gamma}$ expression on gastric cancer have not been reported yet. In this study, we investigated the role of $PPAR-{\gamma}$ expression in gastric cancer patients. Materials and Methods: One hundred twenty-eight (128) samples of both gastric cancer and normal tissues were obtained from 128 patients who had undergone at a curative gastrectomy at Seoul Medical Center from Jan. 2001 to Dec. 2005. $PPAR-{\gamma}$ expression was determined by using immunohistochemical staining, and the results were analyzed. The statistical analysis was based on clinicopathological findings and the differences in survival rates. Results: The mean age of the patients was 6n, and the male : female ratio was 1.9 : 1. $PPAR-{\gamma}$ expression was significantly higher in cancer tissues than in normal tissue (81.3% vs. 57.0%, p<0.001). There was insignificant difference between well and moderately differentiated types and poorly differentiated types in terms of the expression of $PPAR-{\gamma}$ (87.0% vs. 74.6%, P=0.074). In the univariate analysis the survival rate was significantly increased when $PPAR-{\gamma}$ was expressed in normal tissue (P=0.003). In the multivariate analysis, only the UICC TNM staging had significance related to the survival rate. Conclusion: The rate of $PPAR-{\gamma}$ expression was higher in cancer tissue than it was in normal tissue from gastric cancer patients. In the univariate analysis, $PPAR-{\gamma}$ expression in normal tissue had significance with respect to survival, but the multivariate analysis showed no such significance. Thus, we should further evaluate more cases to determine whether or not such a significance exists.

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Correlation between the Serum Leptin Level and the Expression of Leptin in Stomach Cancer Patients (위암에서 혈청 렙틴 레벨과 조직 내 렙틴 발현과의 상관관계)

  • Kim, Ji-Hyun;Jung, Hun;Jun, Kyong-Hwa;Kim, Sung-Keun;Chin, Hyung-Min;Jung, Ji-Han;Kim, Wook;Jeon, Hae-Myung;Park, Cho-Hyun;Park, Seung-Man;Park, Woo-Bae;Lim, Keun-Woo;Kim, Seung-Nam
    • Journal of Gastric Cancer
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    • v.8 no.4
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    • pp.176-181
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    • 2008
  • Purpose: The adipocyte-derived cytokine leptin plays a major role in the control of stable body weight by suppressing food intake and increasing energy metabolism. Leptin regulates the cell proliferation of various epithelial cells and it may be involved in the promotion of cancer. Leptin and its receptor are highly expressed in gastric adenocarcinoma, but the association between the serum leptin level and the tissue expression of leptin is uncertain. We evaluated the serum leptin level and the expressions of leptin and leptin receptor in gastric cancer, and we explore the possible mechanism and role of leptin in the carcinogenesis of gastric cancer. Materials and Methods: 72 carcinomas that were curatively resected at our hospital from October 2005 to March 2007 were included in this study. By immunoassay and immunohistochemical staining, we evaluated the serum leptin level and the expressions of leptin and its receptor, and we analyzed their relationship together with the clinicopathological variables. Results: The serum leptin level was increased as the patient's BMI increased and it was decreased in H. pylori infected patients. The expression of leptin was increased as the TNM stage increased (P=0.014), and the expression of leptin receptor in the intestinal type gastric adenocarcinoma was higher than that in the diffuse type gastric adenocarcinoma (71.4% vs 28.6%, respectively, P=0.033). Conclusion: There was no significant correlation between the serum leptin level and expression of leptin in gastric cancer patients. The expression of leptin was associated with the TNM stage, but its role in the pathogenesis of gastric cancer has to be elucidated.

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The Immunohistochemical Expression of Nuclear Retinoid Receptor and CREB in Gastric Adenocarcinoma (위선암종에서 핵의 Retinoic Acid Receptor (RAR) 및 cAMP Response Element Binding Protein (CREB)의 면역조직화학적 발현양상)

  • Han, Ho-Sun;Kim, Yong-Seok;Park, Joong-Min;Choi, Yoo-Shin;Cha, Seong-Jae;Kim, Mi-Kyung;Chi, Kyung-Choun
    • Journal of Gastric Cancer
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    • v.8 no.4
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    • pp.182-188
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    • 2008
  • Purpose: Transcriptional factors of CREB (cAMP response element binding protein) are involved in regulating the gene expression in response to a variety of signaling pathways. The proteins produced by the CREB genes play key roles in many physiological processes, including memory and long-term potentiation. The retinoic acid receptor (RAR) axis mediates epithelial cell differentiation and proliferation in many tissues. This study examined the expressions of RAR and CREB and their relationship with the clinicopathologic factors and their significance. Materials and Methods: The levels of the RAR and CREB expressions were measured in 150 gastric adenocarcinomas by performing immunohistochemical staining. Results: 1. An RAR protein expression was found in 63.3% of the adenocarcinomas (95/150) and a CREB expression was found in 60.7% (91/150) of the adenocarcinomas. 2. An RAR protein expression was found in 72.2% (78/108) of the intestinal type adenocarcinomas and in 40.5% (17/42) of the diffuse type adenocarcinomas (P<0.05). Based on the depth of invasion, an RAR protein expression was found in 58.3% (14/24) of the T1 adenocarcinomas, in 61.9% (13/21) of the T2 adenocarcinomas, in 63.5% (61/96) of the T3 adenocarcinomas, in 77.8% (7/9) of the T4 adenocarcinomas and in 74.7% (62/83) of the adenocarcinomas with lymph node metastasis and in 49.2% (33/67) of the adenocarcinomas without lymph node metastasis (P<0.01). 3. A CREB expression was found in 69.4% (75/108) of the intestinal type and in 38.1% (16/42) of the diffuse type (P>0.05). Based on the depth of invasion, a CREB expression was found in 50% (12/24) of the T1 adenocarcinomas, in 52.4% (11/21) of the T2 adenocarcinomas, in 64.6% (62/96) of the T3 adenocarcinomas, in 66.6% (6/9) of the T4 adenocarcinomas, in 71.1% (59/83) of the adenocarcinomas with lymph node metastasis and in 47.8% (32/67) of the adenocarcinomas without lymph node metastasis (P<0.01). 4. The RAR protein and CREB expressions coincided in 71.4% of the gastric adenocarcinomas and a significant correlation between them was found (P<0.05). Conclusion: We found a significant relationship between the expression of RAR and CREB and the histology and lymph node metastasis of gastric cancer. Further studies are needed to confirm their biologic meaning in gastric carcinogenesis.

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The Effects of Green Tea on the Serum Lipid and Liver Tissue of Cholesterol Fed Rats (녹차의 음용이 지방과 콜레스테롤을 투여한 흰쥐의 혈청지질 및 간의 조직에 미치는 영향)

  • Sin, Mee-Kung;Han, Sung-Hee;Han, Gun-Jo
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1255-1263
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    • 1997
  • This study was performed to investigate the effects of green tea on fat metabolism of rats and prevention to cardiovascular disease in drinking green tea. Male Spague-Dawley rats were divided into seven groups consisting of the control, lard and cholestrol, aqueous green tea at the level of 1% and 3%. After 4 weeks of feeding serum lipid levels were measured for experimental rats, and analyzed the total cholesterol (TC), HDL-cholesterol (HDL-C), triglyceride (TG), phospholipid (PL). And total lipid (TL) to Folch method, lipid deposition to oil red O staining on liver tissue. The results are as follows: Total cholesterol (TC) decreased by administration of 1% aqueous green tea group and increased addition to lard and cholesterol (LC) group as compared to each groups (p<0.05). HDL-cholestrol in serum increased by administration of la aqueous green tea group (1G) and decreased to the control group, 1% aqueous green tea (L-1G) added lard group (p<0.05). Triglyceride (TG) decreased by administration of 3% aqueous green tea groups (L-3G, LC-3G) and increased by lard and cholesterol group (LC) (p<0.01). Phospholipid(PL) decreased by administration of 3% aqueous green tea added lard and cholesterol group (LC-3G) and increased by control group, lard and cholestrol group (LC) (p<0.05). Total lipid of liver decreased by administration of aqueous green tea at the level of 1% group and increased by LC group (p<0.01). The fat deposition of liver was increased in fat diet groups and decrease in the drink green tea of some but did not showed significant differences from concentration of the green tea.

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EVALUATION OF THE VIABILITY OF PERIODONTAL LIGAMENT CELL IN RAT TEETH USING SLOW CRYOPRESERVATION METHOD WITH MAGNETIC FIELD (자기장 저속 냉동보관법을 이용한 쥐 치아 치주인대세포의 활성도 검사)

  • Ahn, Hyun-Jung;Kim, Eui-Seong;Kim, Jin;Kim, Duck-Won;Kim, Ki-Yeol;Lee, Chan-Young;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.33 no.4
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    • pp.332-340
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    • 2008
  • The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4$^{\circ}C$ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Study on the Development and Properties of Ceramics Restoration Purpose Urethane resin with Excellent Reversibility (가역성이 우수한 도자기 복원용 Urethane resin의 개발 및 물성에 대한 연구)

  • Oh, Seung Jun;Park, Gi Jung;Wi, Koang Chul
    • Journal of Conservation Science
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    • v.31 no.1
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    • pp.37-46
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    • 2015
  • In the case of adhesives & restoration agents currently being used for the preservation treatment of ceramics and earthenware, epoxy type, cellulose type and cellulose type are mainly being used. However, they are showing various problems such as re-detachment from severe contraction, color change from yellowing, work inconvenience of staining on tools and hand during usage and irreversibility. For the purpose of solving the issues of yellowing and irreversibility of epoxy resin being used to restore ceramics, urethane synthetic resin with low yellowing excellent reversibility has been developed in this study. The adhesive strength of urethane resin that has been developed has excellent properties with 2.07MPa for undiluted solution, which is 1.5 times higher than that of existing material EPO-$TEK301^{(R)}$ 1.21MPa. The result of workability measurement showed that the wear rate of urethane resin (in Talc 50wt%) was 1.09%, which was somewhat higher than that of existing material Quick $Wood^{(R)}$ (1.02%). In addition, its wear rate is two times higher than that of $EPO-TEK301^{(R)}$ (0.41%) and $L30^{(R)}$ (0.39%), thereby showing an advantage in its forming process compared to existing materials. As for the advantage of urethane resin of reversibility experiment, 12 hours after acetone, ethyl alcohol deposition, urethane resin and filler talc were dissolved 100% while showing powdering phenomenon. Compared to 0% reversibility of existing epoxy resin, it has much superior reversibility. The result of UV rays experiment to evaluate its durability showed that ${\Delta}E^*ab$ color change value based on undiluted solution of urethane resin was 2.76 before & after UV rays exposure, which was a decrease by about 7-20 times compared to that of existing resin, thereby minimizing the issue of heterogeneity.

Genetic Variations in Geographic Venus Clam(Gomphina aequilatera, Sowerby) Populations from Samcheok and Wonsan (삼척과 원산의 지리적 민들조개(Gomphina aequilatera, Sowerby) 집단의 유전적 변이)

  • Kim, Jong-Rae;Jung, Chang-Ho;Kim, Yong-Ho;Yoon, Jong-Man
    • Development and Reproduction
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    • v.10 no.4
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    • pp.227-238
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    • 2006
  • Genomic DNAs(gDNAs) were isolated from the venus clam(Gomphina aequilatera) from Samcheok(venus clam from Samcheok; VCS) and Wonsan(venus clam from Wonsan; VCW) located in the East Sea of the Korean Peninsula. The amplified products were generated by agarose gel electrophoresis(AGE) with oligonucleotides primer, detected by staining with ethidium bromide and viewed by ultraviolet ray. The seven arbitrarily selected primers BION-21, BION-23, BION-25, BION-27, BION-29, BION-31 and BION-33 generated the shared loci, polymorphic, and specific loci, with the molecular sizes ranging from 150 bp to 2,400 bp. In this study, 147 polymorphic loci(147/954 loci, 15.41%) in VCS population and 274(274/996 loci, 27.51%) in VCW population were generated with seven primers. These results suggest the genetic variation in VCW population is higher than in VCS population. Especially, the 700 bp bands generated by the primer BION-21 were identified commonly in two Gomphina populations, which identified populations and/or species. This specific primer was found to be useful in the identification of individuals and/or population, resulting from the different DNA polymorphism among individuals/species/population. Two Gomphina populations between the individual SAMCHEOK no. 03 and WONSAN no. 22 showed the longest genetic distance(0.696) in comparison with other individuals used. The complete linkage cluster analysis indicating three genetic groupings and dendrogram revealed close relationships among individual identities within two geographical populations of venus clam(G. aequilatera) from the Samcheok and Wonsan. The intra-species classification and clustering analyses inferred from molecular markers supported the traditional taxonomy of the species based on morphological characters such as shell size, shape and color. Accordingly, as mentioned above, RAPD analysis showed that VCS population was more or less separated from VCW population.

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Effects of Different Media and Oxygen Concentrations on In Vitro Maturation and Development of Porcine Follicular Oocytes (배양액과 산소농도가 돼지난포란의 체외성숙과 배발달에 미치는 영향)

  • 천행수;한만희;김종화;박병권;이규승;서길웅
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.119-126
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    • 2004
  • The present study was carried out to examine the effect of four different media (NCSU (North Carolina State University)-23, PZM (Porcine Zygotes Medium)-3, PZM-4 and TCM (Tissue Culture Medium)-l99) and two oxygen concentrations (39 , 5% $O_2$, 5% $CO_2$ and 90% $N_2$, 5% $CO_2$ in air) on in vitro production of porcine IVM/IVF embryos. The results were summarized as follows: The rates of GVBD and nuclear maturations were not significantly different (p>0.05) for 44 hours of culture with four media in two oxygen concentrations. The rates of polyspermy, penetrated sperm(s) and male and female prouclei formation were not significantly different (p>0.05). among four media in two oxygen concentrations. The cleavage rates were not significantly different (p>0.05) among four media in two oxygen concentrations. At day 7 under gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$, the blastocyst formation was significantly higher (p<0.05) in PZM-3 (19.9$\pm$2.4) than other media. Also, NCSU-23 medium gave high rate of blastocyst formation at day 7 under gas atmosphere of 5% $CO_2$ in air (p<0.05). Based on the result of differential staining of porcine blastocyst at dat 7, inner cell mass cell and total cell numbers were not significantly different (p>0.05) among four media in two oxygen concentrations. However, the observed total cell number was higher in PZM-3 medium (36.8$\pm$6.5) than other madia. In conclusion, these results suggested that in vitro production of porcine embryos in PZM-3 medium under a gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$ was effective on the blastocyst formation rate and total blastocyst cell number.