To identify the treatment effect of lactic acid bacteria for diabetes, the treatment effects of a single administration of acarbose (a diabetes treatment drug) or lactic acid bacteria, and the mixture of acarbose and lactic acid bacteria on diabetes in a type 1 diabetes animal model, were studied. In this study, streptozotocin was inoculated into a Sprague-Dawley rat to induce diabetes, and sham control (Sham), diabetic control (STZ), STZ and composition with live cell, STZ and composition with heat killed cell, STZ and composition with drugs (acarbose) were orally administered. Then the treatment effect on diabetes was observed by measuring the body weight, blood glucose, and serum lipid. For the histopathological examination of the pancreas, the Langerhans islet of the pancreas was observed using hematoxylin and eosin staining, and the renal cortex, outer medullar, and inner medullar were also observed. The induced diabetes decreased the body weight, and the fasting blood glucose level decreased in the lactic-acid-bacteriaadministered group and the mixture-administered group. In addition, the probiotic resulted in the greatest decrease in the serum cholesterol level, which is closely related to diabetes. Also, the hematoxylin and eosin staining of the Langerhans islet showed that the reduction in the size of the Langerhans islet slowed in the lactic-acid-bacteria-administered group. The histopathological examination confirmed that the symptoms of diabetic nephropathy decreased in the group to which viable bacteria and acarbose were administered, unlike in the group to which dead bacteria was administered. The mixture of lactic acid bacteria and acarbose and the single administration of lactic acid bacteria or acarbose had treatment effects on the size of the Langerhans islet and of the kidney histopathology. Thus, it is believed that lactic acid bacteria have treatment effects on diabetes and can be used as supplements for the treatment of diabetes.

형질 전환 동물 생산에는 조직 및 시기 특이적 발현 조절이 가능하다는 장점 때문에 유즙 내로 외부 유전자를 발현시키는 시스템이 널리 이용되고 있다. 유전자 발현 즉, 단백질 생산은 프로모터의 강도뿐만 아니라 mRNA의 안정성에 의해서도 조절된다. 특히, polyadenylation에 의한 poly A의 길이는 in vivo와 올 in vitro에서 mRNA 안정성 및 목적 유전자의 번역효율에 영향을 준다. 본 연구에서는 이러한 mRNA 안정성이 목적 유전자의 발현에 미치는 영향을 알아보기 위해 3'-UTR 염기 서열을 분석하였다. 이 3'-untranslated region(UTR) 내의 poly A signal을 기준으로 putative cytoplasmic polyadenylation element(CPE) 부위와 downstream elements(DSE: U-rich, G-rich, GU-rich)의 염기 서열을 분석하고, 각각의 element를 기준으로 15 종의 luciferase reporter vector를 제작하여, 생쥐 유선 세포주(HC11)와 돼지 유선 세포주(PMGC)에 각각 transfection시킨 후 48시간 동안 배양하고 luciferase 발현량을 분석하였다. PMGC의 경우, luciferase의 발현은 exon 9의 CPE 2,3 및 DSE 1을 포함한 #6 construct에서 유의적으로 높은 발현량을 보였으며, exon 9의 CPE 2, 3과 DSE를 모두 포함하고 있는 #11 construct에서도 유의적으로 높은 발현량을 보였다. 이러한 결과는 형질 전환 돼지 생산에 있어 #6 및 11 construct의 사용은 목적의 유전자를 효과적으로 발현시키는데 기여할 것으로 사료된다.

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma 17B-estradiol (E2) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and E2 level were measured by Hemavet 950 and E2 ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma E2 level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma E2 level, thereby causing reproductive disorders.

Transforming growth factor-β (TGF-β) has been shown to have a positive effect on in vitro fertilization (IVF) and has been reported to stimulate meiosis at follicular level in variety of species. The study was designed to determine the expression patterns of TGF-β1, TGF-β receptors type Ⅰ, Ⅱ and Smads gene in bovine oocytes and embryos. TGF-β1 and their receptors were observed in the unfertilized oocytes. TGF-β1 and type Ⅱ receptor were not expressed at the blastocyst stage, however, only type I receptor was exclusively observed at the same stage. The blastocyst stage, in particular, showed high levels of mRNA expression patterns containing a TGF-β type Ⅰ receptor. The mRNA expression pattern of Smad 2 at all stages of embryonic development was similar in all respect with TGF-β1 type I receptor. On the contrary, Smad 3 and 4 were expressed with high and low level mRNA at the blastocyst stage. In conclusion, it is suggested that TGF-β signaling may be regarded as an important entity during the preimplantation embryo development.

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.