• Title/Summary/Keyword: tissues.

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Effects of Cytokinin and Auxin on Organ Formation in Leaf Scale Tissue of Allium Sativum L. (Cytokinin과 Auxin이 Allium Sativum L.의 인엽배양(鱗葉培養)에서 기관(器官)의 분화(分化)에 미치는 영향(影響))

  • Park, Kyeong Han;Lee, Yeong Bok
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.65-76
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    • 1980
  • In order to determine the effects of cytokinin and auxin on organ formation from tissue of garlic cloves, leaf blades and basal tissues contained meristem of garlic (Allium sativum L.) cloves harvested in 1979 (old cloves) and 1980 (new cloves) were explanted on a MS medium contained various levels of BA ($N^6$-benzyl amino purine), NAA (naphthalene acetic acid), and 2, 4-D (2, 4-dichlorophenoxyacetic acid). And some of the new cloves were explanted on a media contained BA and NAA after chilling treatment at $4^{\circ}C$ for 10, 20, 30 and 40 days. 1. In a culture of leaf blades of old cloves, shoots were differentiated on a medium supplemented with 2mg/l of BA and NAA. 2. Callus was grown as a quite straw-coloured globular mass on a medium contained 0.2 or 2mg/l 2.4-D. 3. As subcultures of globular calli, shoots and roots were differentiated on a medium contained 2mg/l BA and 0.5 or 1 mg/l NAA, whereas no shoots was shown on a conterol. 4. Shoots were differentiated in a culture of leaf blades of new cloves, but they were not in an old cloves in control, and better effect was shown on a medium contained 2mg/l BA and 1mg/l NAA. However shoots were no differentiated from leaf blades chilled at $4^{\circ}C$ for 30 or 40 days at the same condition. 5. Large numbers of adventitious shoots could be obtained from basal region of garlic cultured on a medium contained 1mg/l BA and 4mg/l NAA, or 2mg/l BA and 2mg/l NAA.

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Effects of Propylthiouracil and Thyroxine on Thyroid Function and Body Growth in Rats (Propylthiouracil 및 Thyroxine의 처리(處理)가 흰쥐의 갑상선기능(甲狀腺機能)과 체성장(體成長)에 미치는 영향(影響))

  • Seo, Kil Woong;Lee, Kyu Seung;Park, Chang Sik
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.210-218
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    • 1986
  • This study was conducted to find out the effects of propylthiouracil and thyroxine on thyroid function and body growth in female rats. One hundred and forty-four female rats (Wistar-imamichi albino rats) of 25 days old were divided into 4 groups; thyroidectomy (Thx), propylthiouracil (PTU), thyroxine (Thyro.)and control (Cont.) groups. Thirty-six rats were allotted to each group, and changes of body weights were weekly checked. In addition, 6 rats in each group were sacrificed at 1, 2, 3, 5 and 6 weeks after treatments with time elapse for investigating changes of thyroid weights and tissues. The results obtained were as follows: 1. The weights of thyroid gland showed significant differences among all the compared groups at all observation times. The weights of thyroid glands in PTU group were higher than those in control group, but those in the Thyro. group were lower than those in control group. 2. In the histological changes of thyroid glands in the PTU group, the follicle epithelium showed columnar cells following hype trophy and hyperplasia from 1 week after treatment. The follicle epithelium in the Thyro. group were recognized inactive showing squamous cells. 3. The body weights showed significant differences among the compared groups from 2 weeks after treatment. The body weights decreased significantly in PTU and Thx. groups, while those in Thyro. group increased significantly in comparison with those in control group. No significant difference in body weight was noted between PTU and Thx. groups.

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Effects of Probiotic Complex on Performance, Blood Biochemical and Immune Parameters, Digestive Enzyme Activity, Fecal Microbial Population and Noxious Gas Emission in Broiler Chicks (복합생균제가 육계의 생산성, 혈액생화학성분과 면역지표, 소화효소 활성도, 분중 미생물 및 유해가스 발생에 미치는 영향)

  • Kim, Min-Jeong;Jeon, Dong-Gyung;Ahn, Ho-Sung;Yoon, Il-Gyu;Moon, Eun-Seo;Lee, Chai-Hyun;Lim, Yong;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.47 no.3
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    • pp.169-180
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    • 2020
  • This study examined the effects of a probiotic complex (PC) containing Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae on growth performance, organ weight, immune parameters, fecal microbial count, and noxious odor in broiler chicks. A total of 216 birds (4-day-old) were fed a basal diet (CON) and a diet supplemented with 0.25% (PC1) and 0.5% (PC2) of PC until 35 days of age. No difference in body weight, feed intake, and FCR was observed among the groups. The intestinal mucosal weight of the PC1 group was greater than that of the CON group without affecting weights of the other organs. Intestinal secretory immunoglobulin A (sIgA) levels in the PC2 group increased significantly (P<0.05) compared with that in the CON group. The PC2 group also had a strong tendency for elevated blood sIgA levels. Dietary PC did not affect the level of interleukin-1β in the blood and mucosal tissues or alter maltase, sucrase, and leucine aminopeptidase activities in the intestinal mucosa. The PC2 group had higher colony-forming units (cfu) for L. plantarum and S. cerevisiae, but lower cfu for E. coli than those in the CON group. Compared to the CON diet, the PC2 diet resulted in a decreased H2S concentration and a tendency toward decreased CH3SH concentration. In conclusion, a 0.5% PC diet showed increased sIgA and desirable microbial population, and decreased noxious odor in the feces, suggesting that PC could be applied as an environmentally friendly feed additive in broiler chicks.

Chondrogenesis of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood (사람 제대혈 유래 간엽줄기세포로부터 연골세포 분화)

  • Koh, Phil-Ok;Cho, Jae-Hyun;Nho, Kyoung-Hwan;Cha, Yun-Im;Kim, Young-Ki;Cho, Eun-Hae;Lee, Hee-Chun;Jung, Tae-Sung;Yeon, Seong-Chan;Kang, Kyung-Sun;Lee, Hyo-Jong
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.528-533
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    • 2009
  • In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.

Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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Immunohistochemical Study on the Activation of Cell mediated immunity in Murine Lymph node on Allergic Contact Dermatitis by DNCB -Based on the change of T lymphocytes and Il-2 receptors- (알러지성 접촉피부염 유발 피부 주변 림프절에서의 세포성 면역 활성에 관한 면역조직화학적 연구 - T 림프구와 IL-2 수용기의 분포 변화를 중심으로 -)

  • kim, Jin-Taek;Ahn, Sang-Hyun;Park, In-Sick;Chung, Jae-Man;Kim, Ho-Hyun
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.1
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    • pp.33-41
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    • 1998
  • Lymph node tissues of BALB/C mouse treated with DNCB were immunohistochemically observed to investigate the activation of cell mediated immunity in lymph node of murine with allergic contact dermatitis. The inguinal region of BALB/C mice were sensitized by one application of $25{\mu}l$ of 5% 2,4-dinitrochlorobenzene(DNCB) onto an abdominal skin and 2 weeks later, the mice were challenged with $4{\mu}l$ of 2.5% DNCB. The inguinal lymph node were obtained at hour 24, 48, and 72 after 2nd DNCB treatment and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25). The distribution of helper T lymphocytes, cytotoxic T lymphocytes and IL-2 receptors began to increase at hour 24 after after 2nd DNCB treatment and these increase appeared in paracortical area and medullary sinius. These increase were greatest at hour 48. These results indicated that the IL-2 secretion began to increase by activation of helper T lymphocytes in lymph node of DNCB re-exposure area and subsequently to activate suppress T lymphocytes.

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Establishment of a Single Dose Radiation Model of Oral Mucositis in Mice (일회 방사선조사를 이용한 마우스 구강점막염 모델의 확립)

  • Ryu, Seung-Hee;Moon, Soo-Young;Choi, Eun-Kyung;Kim, Jong-Hoon;Ahn, Seung-Do;Song, Si-Yeol;Park, Jin-Hong;Noh, Young-Ju;Lee, Sang-Wook
    • Radiation Oncology Journal
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    • v.26 no.4
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    • pp.257-262
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    • 2008
  • Purpose: Oral mucositis induced by radiotherapy to the head and neck area, is a common acute complication and is considered as the most severe symptom for cancer patients in the early stages of treatment. This study was proposed to establish the oral mucositis mouse model induced by a single dose of radiation for the facility of testing therapeutic candidates which can be used for the oral mucositis treatments. Materials and Methods: Fifty-five BALB/c mice were divided into four groups: control, 16 Gy, 18 Gy, and 20 Gy. Oral mucositis was induced by a single dose of radiation to the head and neck using 6 MV x-Ray from linear accelerator. After irradiation, body weight and physical abnormalities were checked daily. Tongue tissues from all groups were taken on days 1, 2, 3, 5, 7, 9, and 14, respectively and H&E staining was conducted to examine morphological changes. Results: Body weight dramatically decreased after day 5 in all irradiated mice. In the 16 Gy treatment group, body weight was recovered on day 14. The histology data showed that the thickness of the epithelial cell layer was decreased by the accumulated time after radiation treatment, up to day 9. Severe ulceration was revealed on day 9. Conclusion: A single dose of 16 Gy is sufficient dose to induce oral mucositis in Balb/C mice. Significant changes were observed in the Balb/C mice on days 7 and 9 after radiation. It is suggested that this mouse model might be a useful standard tool for studying oral mucositis induced by radiation.

The Efficacy of Needle-Acupuncture and Moxibustion at BL18 on Hepatic Injury in Dogs (간유(肝兪)(BL18)에 대한 자침 및 뜸 요법의 개 간 손상 회복에 미치는 효과)

  • Jun, Hyung-Kyou;Lee, Chang-Seok;Lee, Jung-Yeon;Lee, Sang-Eun;Song, Kun-Ho;Park, Bae-Gun;Cho, Sung-Whan;Kim, Duck-Hwan
    • Journal of Veterinary Clinics
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    • v.24 no.1
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    • pp.10-14
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    • 2007
  • The present study was performed to elucidate the recovery effect of needle-acupuncture (needle-AP) and moxibustion treatments at Gan Shu (BL18) on $CCl_4$ induced hepatic injury in dogs, Total 9 clinically healthy experimental dogs (1 to 2 years old, 2.3-5.3kg body weight) were divided into control (3 dogs), needle-AP (3 dogs) and moxibustion (3 dogs) groups, respectively. Hepatic iniw was induced by intraperitoneal injection with $CCl_4$ (olive oil : $CCl_4=1:1,\;1ml/kg$, once/day). As far the treatments in each group, control group was not treated at all after induction of the hepatic injury till the end of experiment. Needle-Af group was treated by AP at BL18 (once/day, 20 minutes) from the next day of induction of the hepatic injury for 11 days. Moxibustion group was also treated with commercial moxa at BL18 (once/day, 20minutes) for 11days. The changes of the serum alanine transaminase(ALT), aspartate transaminase(AST) and gamma glutamyl transpeptidase (GGT) activities were investigated on pre, 0, 1, 3, 5, 7 and 11 days after hepatic injury, respectively. Histopathological changes were also investigated in the liver tissues on day 11 in experimental and control groups. The results obtained in the present study were as follows. In needle-AP group, they showed significant lower values on the 1st (p<0.05) and $3^{rd} day (p <0.05)$ in serum ALT activities, and the $11^{th}$ day in serum GGT activities, compared with those of control group, respectively. However, the significances were not detected in serum AST activities of needle-AP group by comparison with those of control group. In moxibustion group, they showed significant low values on the $1^{st}(p<0.05)$, 3rd day (p<0.05) and $5^{th}$ day (p<0.05) in serum ALT activities, compared with those of control group, respectively. However, significances were not detected in serum GGT and AST activities of moxibustion group by comparison with those of control group. Among the experimental groups, the significant low values were found on $1^{st}\;and\;3^{rd}$ day in serum ALT activities (p<0.05), and $3^{rd}\;and\;5^{th}$ day in serum GGT activities (p<0.05) in needle-AP group, compared by those of moxibustion group, respectively. In addition, the marked recovery findings of histopathological changes in experimental groups were found, compared by those of control group: mild vacuolar degeneration and necrosis findings in needle-AP group, and moderate vacuolar degeneration and necrotic findings in moxibustion group were found, in contrast to severe changes such as accumulation of bile juice, vacuolar degeneration and necrotic findings observed in control group. In conclusion, needle-AP and moxibustion treatments at BL-18 were effective for the recovery for $CCl_4$ induced hepatic injury and needle-AP was more effective than that of moxibustion in dogs.

Development of a Simple and Reproducible Method for Removal of Contaminants from Ginseng Protein Samples Prior to Proteomics Analysis (활성탄을 이용한 불순물제거에 의한 효과적인 인삼 조직 단백질체 분석 방법 개선 연구)

  • Gupta, Ravi;Kim, So Wun;Min, Chul Woo;Sung, Gi-Ho;Agrawal, Ganesh Kumar;Rakwal, Randeep;Jo, Ick Hyun;Bang, Kyong Hwan;Kim, Young-Chang;Kim, Kee-Hong;Kim, Sun Tae
    • Journal of Life Science
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    • v.25 no.7
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    • pp.826-832
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    • 2015
  • This study describes the effects of activated charcoal on the removal of salts, detergents, and pigments from protein extracts of ginseng leaves and roots. Incubation of protein extracts with 5% (w/v) activated charcoal (100-400 mesh) for 30 min at 4℃ almost removed the salts and detergents including NP-40 as can be observed on SDS-PAGE. In addition, analysis of chlorophyll content showed significant depletion of chlorophyll (~33%) after activated charcoal treatment, suggesting potential effect of activated charcoal on removal of pigments too along with the salts and detergents. 2-DE analysis of activated charcoal treated protein samples showed better resolution of proteins, further indicating the efficacy of activated charcoal in clearing of protein samples. In case of root proteins, although not major differences were observed on SDS-PAGE, 2-DE gels showed better resolution of spots after charcoal treatment. In addition, both Hierarchical clustering (HCL) and Principle component analysis (PCA) clearly separated acetone sample from rest of the samples. Phenol and AC-phenol samples almost overlapped each other suggesting no major differences between these samples. Overall, these results showed that activated charcoal can be used in a simple manner to remove the salts, detergents and pigments from the protein extracts of various plant tissues.

Potential Role of Hedgehog Signaling in Radiation-induced Liver Fibrosis (방사선에 의한 간섬유증에서 헤지호그의 잠재적 역할)

  • Wang, Sihyung;Jung, Youngmi
    • Journal of Life Science
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    • v.23 no.5
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    • pp.710-720
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    • 2013
  • Radiotherapy is commonly used in treating many kinds of cancers which cannot be cured by other therapeutic strategies. However, radiotherapy also induces the damages on the normal tissues. Radiation-induced fibrosis is frequently observed in the patients undergoing radiotherapy, and becomes a major obstacle in the treatment of intrahepatic cancer. Hedgehog (Hh) that is an essential in the liver formation during embryogenesis is not detected in the healthy liver, but activated and modulates the repair process in damaged livers in adult. The expression of Hh increases with the degree of liver damage, regulating the proliferation of hepatic progenitors and hepatic stellate cells (HSC). In addition, Hh induces epithelial-to-mesencymal transition (EMT) and activation of myofibroblasts. In the irradiated livers, up-regulated expression of Hh signaling was associated with proliferation of progenitors, EMT induction, and increased fibrosis. Female-specific expression of Hh leaded to the expansion of progenitors and the accumulation of collagen in the irradiated livers of female mice, indicating that gender disparity in Hh expression may be related with radiation-susceptibility in female. Hence, Hh signaling becomes a novel object of studies for fibrogenesis induced by radiation. However, the absence of the established experimental animal models showing the similar physiopathology with human liver diseases and fibrosis-favorable microenvironment hamper the studies for the radiation-induced fibrosis, providing a few descriptive results. Therefore, further research on the association of Hh with radiation-induced fibrosis can identify the cell and tissue-specific effects of Hh and provides the basic knowledge for underlying mechanisms, contributing to developing therapies for preventing the radiation-induced fibrosis.