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Quality Assay of Human Fetal-Cord Serum for Human IVF-ET with Mouse 2-Cell Embryos (생쥐 2-세포배아에 의한 시험관아기 배양용 대아제대혈청의 절적평가에 관한 연구)

  • Moon, S.Y.;Shin, C.J.;Chung, K.M.;Oh, S.K.;Pang, M.G.;Chang, Y.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.2
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    • pp.139-146
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    • 1989
  • The purpose of this study was to examine the qualitative variation of human fetal-cord sera (HCS) and to accept the sera in human lVF-ET program. One hundred and sixteenth RCS were tested with 1772 2-cell embryos of F1 (C57BL x CBA) virgin mice, Ten to sixteenth embryos were cultured in m-KRB medium with a aliquot of each serum (10%, v/v) or with bovine serum albumin(O.4%, w/v) as a control medium. Embryonic development were recorded at every 24hr for 4 days by such events as cellular compaction, cavitation, and hatching. In the control groups of eight assays, 98.1%(106/ 108) of 2-ce1l embryos developed above expanded blastocyst and the embryonic development was unified through the tests. But the developmental pattern in medium with each serum was various. Namely, the sera that supported development of 100% 2-cell embryos to above morula, early blastocyst, expanded blastocyst and hatching blastocyst was 45,7%(53/116) , 35.3%(41/116), 15.5%08/116.) and 6.9-%(8/116), respectively. And the sera that supported development of above 80% 2-cell embryos to the each embryonic stage was 92.2% (107/116), 83.6%(97/116), 63.8%(74/116) and 36.2%(42/116), respectively. Meanwhile two kinds of toxic pattern to the embryonic development were observed in some sera. The first pattern is that some sera arrested development of most embryos in pre- or post-stage of morula or blastocyst. The second pattern is that some sera promoted or arrested a part of embryos in the same dish. The ability of serum was depended on the batch of serum. Finally we could accept 69%(80/116) of the tested sera for human IVF-ET program. The base line for acceptance was the ability that supported above 80% 2-ce1l embryos to blastocyst. But some deterious sera were contained in this range. We cut off about 10% of the sera (83.6% , 97/116) that passed the baseline. This final percent of sera was similar to that of grade N of this study.

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Polybrominated Diphenyl Ethers Orally Administration to Mice Were Tansferred to Offspring during Gestation and Lactation with Disruptions on the Immune System

  • Hong, Soon-Keun;Sohn, Kyung-Hee;Kim, In-Young;Lee, Jong-Kwon;Ju, Jung-Hun;Kim, Jin-Ho;Lim, Chae-Hyung;Han, Beom-Seok;Jung, Hwa-Chul;Lee, Jin-Yong;Park, Kui-Lea
    • IMMUNE NETWORK
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    • v.10 no.2
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    • pp.64-74
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    • 2010
  • Background: The present study was undertaken to examine the immunological effects of pentabrominated diphenyl ether (penta-BDE) and decabrominated diphenyl ether (deca-BDE) on the immune system of the dams and the developmental immune system of the offsprings. Methods: In this study, mated female C57BL/6J mice were orally administered penta-BDE, deca-BDE or corn oil for 5 weeks, from gestational day 6 to lactational day 21. Results: The body weight of PND21 exposed to penta-BDE was significantly decreased relative to control mice, but that of post-natal day 63 (PND63) were recovered. Orally dosed dams with penta-BDE had significantly smaller absolute and relative spleen masses than control mice. Absolute and relative spleen and thymus masses of PND21 exposed to penta-BDE were significantly decreased over control. The exposure of dams and PND21 with penta-BDE reduced the number of splenocytes and thymocytes. As results of hematologic analysis, percentage WBC and percentage neutrophils increased in dams with deca-BDE. Splenic T cell proliferation in dams and PND21 exposed to penta-BDE was increased, and there were no significant difference in splenic B cell proliferation in all treatment groups. As results of flow cytometric analysis of splenocyte, percentage total T cell, Th cell and Tc cell in PND21 exposed to penta-BDE was slightly increased, and percentage macrophage in dams and PND21 exposed to deca-BDE was decreased. The ELISA results of antibody production show no significant difference in all treatment groups relative to controls. Conclusion: These results imply that PBDEs given to the dam were transferred to the offspring during gestation and lactation, and PBDEs transferred from the dam affect immune system of offspring.

In Vitro Differentiation-induced hES Cells Relieve Symptomatic Motor Behavior of PD Animal Model

  • 이창현;김은경;이영재;주완석;조현정;길광수;이금실;신현아;안소연
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.95-95
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    • 2002
  • Human embryonic stem (hES) cells can be induced to differentiate into tyrosine hydroxylase expressing (TH+) cells that may serve as an alternative for cell replacement therapy for Parkinson's disease (PD). To examine in vitro differentiation of hES (MB03, registered in NIH) cells into TH+ cells, hES cells were induced to differentiate according to the 4-/4+ protocol using retinoic acid (RA), ascorbic acid (AA), and/or lithium chloride (LiCl) followed by culture in N2 medium for 14 days, during which time the differentiation occurs. Immunocytochemical stainings of the cells revealed that approximately 21.1% of cells treated with RA plus AA expressed TH protein that is higher than the ratio of TH+ cells seen in any other treatment groups (RA, RA+LiCl or RA+AA+LiCl). In order to see the differentiation pattern in vivo and the ability of in vitro differentiation-induced cells in easing symptomatic motor function of PD animal model, cells (2 $\times$ 10$^{5}$ cells/2${mu}ell$) undergone 4-/4+ protocol using RA plus AA without any further treatment were transplanted into unilateral striatum of MPTP-lesioned PD animal model (C57BL/6). Following the surgery, motor behavior of the animals was examined by measuring the retention time on an accelerating rotar-rod far next 10 weeks. No significant differences in retention time of the animals were noticed until 2 weeks post-graft; however, it increased markedly at 6 weeks and 10 weeks time point after the surgery. Immunohistochemical studies confirmed that a reasonable number of TH+ cells were found at the graft site as well as other remote sites, showing the migrating nature of embryonic stem cells. These results suggest that in viかo differentiated hES cells relieve symptomatic motor behavior of PD animal model and should be considered as a promising alternative for the treatment of PD.

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Residual, Unresectable and Recurrent Rectal Cancer : Role of External Radiation Therapy in 46 Patients (국소 재발성 또는 진행된 직장암의 방사선 치료 -46예의 치료 성적 분석-)

  • Gil, Hack-Joon;Oh, Yoon-Kyeong;Yoon, Sei-Chul;Shinn, Kyung-Sub;Bahk, Yong-Whee
    • Radiation Oncology Journal
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    • v.6 no.1
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    • pp.55-61
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    • 1988
  • Fifty patients with residual, unresectable or recurrent rectal cancer were treated with external irradiation using a 6-MV linear accelerator at the Division of Therapeutic Radiology, Department of Radiology, Kangnam St. Mary's Hospital, Catholic University Medical College during the period of April 1983 to December 1987. This paper describes the results of a retrospective analysis of the results of external irradiation for the residual, unresectable and recurrent rectal cancer in 46 patients. Four patients were lost to follow-up. Of the 46 patients, $18 (39\%)$ presented with unresectable primary lesions and $28 (61\%)$ with residual or recurrent rectal cancer. In $93\%$, the pathologic diagnosis was adenocarcinoma. Resonse to irradiation was observed in $22 (73\%)$ out of 30 patients who were treated for pain, $12 (86\%)$ out of 14 patients who were treated for mass, and $17 (77\%)$ out of 22 patients who were treated for bloody discharge. The actuarial postoperative 2-year and 3-year survival rates in recurrent and unresectable patients were $43\%$ and $22\%$, respectively. However, the post-RT 2-year survival rate was $13\% (6/46)$.

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Patterns of Protein Synthesis During the Second Cleavage of Mouse Two-Cell Embryos: Effects of Colcemid and a-Amanitin (생쥐 배아의 2세포기 분열과정에 있어서의 단백질 합성 분석 : Colcemid와 a-Amanitin의 영향)

  • Kang, Hae-Mook;Kvu
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.404-411
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    • 1989
  • In this study, we attempted to determine the precise patterns of protein synthesis during the second cleavage in mouse. F1 hybrid 2-cell embryos showing a highly synchronized cell cycle and outbreed ICR strain 2-cell embryos were used. The patterns of protein synthesis during the second cleavage showed the sequential changes in the F1 hybrid and ICR strain 2-cell embryos. Moreover, we examined the effects of mitotic and transcriptional inhibitors such as colcemid and a-amanitin on the protein synthesis in the late 2-cell embryos of ICR strain. Treatment of colcemid (0.1mg/ml) blocked the second cleavage, but did not affect on the change of protein synthesis. However, treatment of a-amanitin induced the synthesis of two set of polypeptides without affecting on synthesis of other proteins and cleavage. It thus seems that the appearance of a-amanitin-sensitive proteins may be not involved in the second cleavage. Therefore, these results indicate that the second cell cycle in mouse embryos appears to be regulated at post transcriptional level, presumably independent on the expression of embryonic genome. 본 연구는 생쥐 배아의 2세포기 분열과정중 단백질 합성양상과 단백질합성에 미치는 colcemid와 $\alpha$-amanitin의 영향을 조사하였다 이를 위하여 체내 수정된 ICR strain의 2세포기 배아와 매우 일치된 초기배아 분열양상을 보여주는 체외 수정된 F1 (C57BL x CBA) hybrid 2세포기 배아를 사용하였다. 두 종류의 2세포기 배아에서 단백질 합성은 분열단계에 따라서 매우 일치된 변화를 보여 주었다. 또한 유사분열 억제제인 colcemid (0.1mg/ml)의 처리는 2세포기 배아분열을 억제하였으나, 단백질 합성에는 아무런 변화를 주지 못하였다. 그리고 후기 2세포기 배아에 전사 억제제인 a-amanitin (100mg/ml)을 처리하였을 때 세포분열이나 다른 단백질의 합성에는 아무런 영향이 없이 단지 두개의 단백질의 합성만을 유도하였다. 이는 아마도 a-amanitin의 stress효과에 기인하는 것으로 추측된다. 따라서 생쥐 2세포기 배아의 분열과정은 배아게놈의 유전자 발현과는 무관하게 이미 합성되어 존재하는 mRNA에 의하여 조절되는 것으로 사료된다.

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The Clinical Review of Samgi-Halleak Pharmacopuncture Effects for Insomnia & Fatigue (삼기활력약침이 불면, 피로에 미치는 효과)

  • Lee, Yoo-Hwan;Kwon, Gi-Sun;Lee, Seung-Hwon;Lee, Eun-Sol;Kim, Cheol-Hong;Jang, Kyung-Jun;Song, Choon-Ho;Kim, Young-Gyun;Kim, Won-Il;Yoon, Hyun-Min
    • Journal of Acupuncture Research
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    • v.29 no.3
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    • pp.101-113
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    • 2012
  • Objectives : The purpose of this study is to investigate the effects of Samgi-Halleak pharmacopuncture treatment on insomnia and fatigue. Methods : This study was done with 47 nurses under shiftwork schedule. Subjects were divided into two groups ; Samgi-Halleak pharmacopuncture treated group(experimental group, N=24), normal saline treated group(control group, N=23). The procedures had been conducted to the subjects injecting 0.1~0.2mL total 1mL, 0.5~1cm deep on each acupoint 2 times per week. The acupoints were Pungji($GB_{20}$), Gyeonjeong($GB_{21}$), Sinsu($BL_{23}$). Both of groups were treated total 8 times, but control group was treated additional pharmacopuncture the same way as experimental group after 8 times normal saline treatment. The collected data were analyzed with insomnia and fatigue at baseline and post pharmacopuncture using independent samples t-test, Paired t-test, ${\chi}^2$-test by SPSS 18.0 Windows program. Results : After 8 times treatment, the scores of both groups showed significant improvement in insomnia and fatigue. Comparing the experimental and control group, there were more significant improvement in experimental group than control group in the scores of insomnia and fatigue. Conclusions : Samgi-Halleak pharmacopuncture can be used for effective treatment in patients with insomnia and fatigue.

Prospero Homeobox 1 and Doublecortin Correlate with Neural Damage after Ischemic Stroke

  • Dong-Hun Lee;Eun Chae Lee;Sang-Won Park;Ji young Lee;Kee-Pyo Kim;Jae Sang Oh
    • Journal of Korean Neurosurgical Society
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    • v.67 no.3
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    • pp.333-344
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    • 2024
  • Objective : Markers of neuroinflammation during ischemic stroke are well characterized, but additional markers of neural damage are lacking. The study identified associations of behavioral disorders after stroke with histologic neural damage and molecular biological change. Methods : Eight-week-old, 25 g male mice of the C57BL/6J strain were subjected to middle cerebral artery occlusion (MCAO) to induce ischemic stroke. The control group was a healthy wild type (WT), and the experimental group were designed as a low severity MCAO1 and a high severity MCAO2 based on post-stroke neurological scoring. All groups underwent behavioral tests, realtime polymerase chain reaction, triphenyltetrazolium chloride (TTC) staining and Hematoxylin and Eosin staining. One-way analysis of variance was used to analyze statistical significance between groups. Results : In TTC staining, MCAO1 showed 29.02% and MCAO2 showed 38.94% infarct volume (p<0.0001). The pro-inflammatory cytokine interleukin (IL)-1β was most highly expressed in MCAO2 (WT 0.44 vs. MCAO1 2.69 vs. MCAO2 5.02, p<0.0001). From the distance to target in the Barnes maze test, WT had a distance of 178 cm, MCAO1 had a distance of 276 cm, and MCAO2 had a distance of 1051 (p=0.0015). The latency to target was 13.3 seconds for WT, 27.9 seconds for MCAO1, and 87.9 seconds for MCAO2 (p=0.0007). Prospero homeobox 1 (Prox1) was most highly expressed in MCAO2 (p=0.0004). Doublecortin (Dcx) was most highly expressed in MCAO2 (p<0.0001). Conclusion : The study demonstrated that histological damage to neural cells and changes in brain mRNA expression were associated with behavioral impairment after ischemic stroke. Prox1 and Dcx may be biomarkers of neural damage associated with long-term cognitive decline, and increased expression at the mRNA level was consistent with neural damage and long-term cognitive dysfunction.

Effect of Glucose Exposure on the Development of the Mouse Preimplantation Embryo In Vitro (착상전 생쥐배아의 Glucose에 대한 노출이 체외 배발생에 미치는 영향)

  • Kim, Seon-Ui;Eom, Sang-Jun;Yun, San-Hyeon;Im, Jin-Ho;Jeong, Gil-Saeng
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.227-234
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    • 1995
  • This study was carried out to investigate the effects according to the time course of glucose exposure on the development of one-cell embryos beyond morula in CR$_laa$ medium. One-cell zygotes from B6CBA F$_1$ mice were recovered at 24 ~ 25h after hCG and cumulus cells were removed with 0.1% hyaluronidase. The embryos were pooled and subsequently divided into each groups and cultured in CR$_laa$ at 37$^{\circ}C$ in 5% CO$_2$ in aIr. The embryos were either, placed in CR$_laa$ containing various concentration (5.5, 16.5, 27.5 and 38.5 mM) of glucose for 1 min. and subsequently returned to the fresh culture medium (without glucose), or were transferred to the same media containing glucose at 72 h post hCG. The results obtained in these experiments were summarized as follows: 1. The development rates of zygotes, recovered from the oviducts in M2 and cultured in CR$_laa$ with 3mg/Im FAF-BSA, to expanded blastocysts (25.7%) and hatching bIastocysts (17.6%) were significantly higher than those of zygotes recovered in TL Hepes (0% and 0%, respectively). 2. The development rates of one-cell embryos exposed to 27.5 mM glucose at 72 h post hCG for 1 min, were 68.8% (CR$_1$+BSA) a and 77.1% (CR$_1$+FBS) of expanded blastocyst stage, but there were no significant differences between the embryos exposed for 1 min. or transferred at 72 h. 3. Regardless of glucose concentration (5.5, 16.5, 27.5 & 38.5mM), 45.7~61.5% of embryos developed to the blastocyst stage. There were no significant differences between any of the treatments on the devel-opment of one-cell embryos. Therefore, the detrimental effect of highly concentration was not appeared.

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Emergency Coronary Artery Bypass Operation for Card iogen ic Shock (심인성 쇼크에 대한 응급 관상동맥 우회술)

  • 김응중;이원용
    • Journal of Chest Surgery
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    • v.30 no.10
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    • pp.966-972
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    • 1997
  • Between June 1994 to August 1996, 13 patients underwent emergency coronary artery bypass operations. There were 3 males and 10 females and ages ranged from 56 to 80 years with the mean of 65.5 years. The indications for emergency operations were cardiogenic shock in 12 cases and intractable polymorphic VT(ve'ntricular tachycardia) in 1 case. The causes of cardiogenic shock were acute evolving infarction in 6 cases, PTCA failure in 4 cases, acute myocardial infarction in 1 case, and post-AMI VSR(ventricular septal rupture) in 1 case. Pive out of 13 patients could go to operating room within 2 hours. However, the operations were delayed from 3 to 10 hours in 8 patients due to non-medical causes. In 12 patients, 37 distal anastomoses were constructed with only 3 LITA's(left internal thoracic arteries) and 34 saphenous veins. In a patient with post-AMI VSR, VSR repair was added. In a patient with intractable VT and critical sten sis limited to left main coronary artery, left main coronary angioplasty was performed. Pive patients died after operation with the operative mortality of 38.5%. Three patients died in the operating room due to LV pump failure, one patient died due to intractable ventricular tachycardia on postoperative second day, and one patient died on postoperative 7th day due to multi-organ failure with complications of mediastinal bleeding, low cardiac output syndrome, ARF, and lower extremity ischemia due to IABP. In 8 survived patients, 3 major complications (mediastinitis, PMI, UGI bleeding) developed but eventually recovered. We think that the aggressive approach to critically ill patients will salvage some of such patients and the most important factor for patient salvage is early surgical intervention before irreversible damage occurs.

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Immunostimulntory Effects of Immu-Forte at 3 Months Post-Treatment in Mice (면역기능증강성 동암 바이오스 신물질에 대한 3개월간의 마우스 투여후의 면역학적 및 혈액학적 변화)

  • Jung Ji-Youn;Ahn Nam-Shik;Park Joon-Suk;Jo Eun-Hye;Hwang Jae-Woong;Lee Seoung-Hun;Park Jung-Ran;Kim Sun-Jung;Lee Yong-Geon;Jeong Yun-Hyeok;Chung Ji-Hye;Lee Soo-Jin;Lee Sang-Bum
    • Journal of Food Hygiene and Safety
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    • v.20 no.2
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    • pp.118-122
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    • 2005
  • Immu-Forte (Dong-Ahm Bio's. Corp., Korea) was evaluated fir its effectiveness as a nonspecific immunostimulator in mice. The effects of Immu-Forte were determined by analysis of cytokines using ELISh and phenotype of leukocyte subpopulations using monoclonal antibodies specific to mouse leukocyte differentiation antigens and flow cytometry. CD4 T cells, CD8 T cells, macrophages, IL-12 and IFN-r in Immu-Forte EX-treated middle dose group increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. All T cells, all B cells, macrophages, IL-2, IL-4 and IL-12 in Immu-Forte EX-treated low dose uoup increased in 3 months posttreatment and were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte soy-treated group, CD4 T cells, IL-2, IL-4 and IL-12 were significantly higher in high dose-treated group, and CD 4 T cell, macrophages, IL-2, IL-4 and IL-12 were significantly higher in middle dose-treated group, and all T cell, IL-2, IL-4 and IL-12 were significantly higher in low dose-treated group. In the Itnmu-Forte A-treated group, macrophages, m cells and IL-12 in high dose-treated group and all T cells, macrophages, NK cells, IL-2, IL-4 and IL-12 in middle dose-treated group and NK cells in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In the Immu-Forte F-treated Group, all B cells, IL-4 and IL-12 in high dose-treated group and all T cells, aBl B cells, CD 4 T cells, CD8 T cells, macrophage, IL-2, IL-4, IL-12 and IFN-r in middle dose-treated group and NK cells and IL-12 in low dose-treated group were significantly higher (p<0.05) than that of control at 3 months posttreatment. In conclusion, the study has demonstrated that Immu-Forte had an immunostimulatory effect on mice through proliferation and activation of mouse immune cells.