• Title/Summary/Keyword: Gene injection

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Molecular Biologic Study on the Role of Glutamate in Spinal Sensitization (척수통증과민반응에서 Glutamate의 역할에 대한 분자생물학적 연구)

  • Kim, Hae-Kyu;Jung, Jin-Sup;Baik, Seong-Wan
    • The Korean Journal of Pain
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    • v.14 no.1
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    • pp.1-6
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    • 2001
  • Background: Subcutaneous injection of 5% formalin into the hind paw of the rat produces a biphasic nociceptive response. The second phase depends on changes in the dorsal horn cell function that occur shortly after an initial C-fiber discharge, spinal sensitization, or windup phenomenon. This study was performed to investigate the role of glutamate during spinal sensitization. Methods: Sprague-Dawley rats weighing 200 to 250 g were used for this study. Under light anesthesia (0.5% isoflurane) the rats were segregated in a specially designed cage and $50{\mu}l$ 0.5% formalin was injected subcutaneously in the foot dorsum of right hindlimb. Forty minutes after the formalin injection, the rat was quickly decapitated and spinal cord was removed. The spinal segments at the level of L3 (largest area) was collected and stored in a deep freezer ($-70^{\circ}C$). The mRNA gene expression of N-methyl-D-aspartate receptor (NMDAR) and the metabotropic glutamate receptor subtype 5 (mGluR5) were determined by the polymerase chain reaction. Results: The number of flinches was $19.8{\pm}2.3/min$. at one minute after formalin injection and decreased to zero after then. The second peak appeared at 35 and 40 minutes after formalin injection. The values were $17.8{\pm}2.2$ and $17.2{\pm}3.0/min$. The mRNA gene expressions of NMDAR and mGluR5 were increased by $459.0{\pm}46.8%$ (P < 0.01) and $111.1{\pm}4.8%$ (P > 0.05) respectively at 40 minutes after formalin injection. The increased rate of NMDAR was significantly higher than that of mGluR5 (P < 0.01). Conclusions: From these results it suggested that NMDAR partly contributed to the mechanism of central sensitization after the formalin test but mGluR5 did not.

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Therapeutic applications of gene editing in chronic liver diseases: an update

  • Shin, Ji Hyun;Lee, Jinho;Jung, Yun Kyung;Kim, Kyeong Sik;Jeong, Jaemin;Choi, Dongho
    • BMB Reports
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    • v.55 no.6
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    • pp.251-258
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    • 2022
  • Innovative genome editing techniques developed in recent decades have revolutionized the biomedical research field. Liver is the most favored target organ for genome editing owing to its ability to regenerate. The regenerative capacity of the liver enables ex vivo gene editing in which the mutated gene in hepatocytes isolated from the animal model of genetic disease is repaired. The edited hepatocytes are injected back into the animal to mitigate the disease. Furthermore, the liver is considered as the easiest target organ for gene editing as it absorbs almost all foreign molecules. The mRNA vaccines, which have been developed to manage the COVID-19 pandemic, have provided a novel gene editing strategy using Cas mRNA. A single injection of gene editing components with Cas mRNA is reported to be efficient in the treatment of patients with genetic liver diseases. In this review, we first discuss previously reported gene editing tools and cases managed using them, as well as liver diseases caused by genetic mutations. Next, we summarize the recent successes of ex vivo and in vivo gene editing approaches in ameliorating liver diseases in animals and humans.

Intravenous Delivery of Transgene-Liposome Complexes

  • Park, Seung-Kyu;Kim, Sun-Uk;Cho, Na-Na;Park, Chang-Sik;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.187-187
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    • 2004
  • Gene delivery is one of the keen interests in animal industry as well as research on gene function. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. (omitted)

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Parallel Regulation of Prolactin and c-fos Gene Expression by 17$\beta$-estradiol and Stress in the Mouse Pituitary

  • Kim, Ji-Eune;Ko, Ji-Yun;Kim, Young-il;Yoon, Yong-Dal;Cho, Byung-Nam
    • Animal cells and systems
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    • v.4 no.1
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    • pp.71-76
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    • 2000
  • The aim of this study was to investigate expression patterns of the prolactin (PRL) and c-fos genes by 17$\beta$-estradiol (17$\beta$-E) and stress in the mouse pituitary. In the pituitary, the levels of PRL mRNA were found high with some fluctuation at 30, 50, and 90 min whereas the levels of PRL mRNA were low at 120 min when ovariectomized female mice were injected with 17$\beta$-E or vehicle. PRL mRNA levels began to increase again at 4 h and remained high up to 24 h only in the 17$\beta$-E- treated mice. The overall changes in c-fos mRNA by 17$\beta$-E were very similar to those in PRL mRNA in the pituitary. Subsequent study revealed that these high initial levels of PRL and c-fos mRNAs were caused by stress during Injection, not by 17$\beta$-E, since vehicle injection alone into the ovariectomized mice could increase the levels of PRL and c-fos mRNAs. The stress-induced elevations of PRL and c-fos mRNAs were inhibited by bromocriptin, a dopamine agonist, suggesting that the dopaminergic system is involved in the action route of injection stress. In addition, the induced levels of c-fos mRNA by 17$\beta$-E and stress in the pituitary were very low compared with those in the uterus. The time course changes in c-fos mRNA level were different between the pituitary and uterus. Taken together, these data indicate that PRL and c-tos gene expression in the pituitary are regulated by 17$\beta$-E and stress in a parallel manner, supporting the notion that c-Fos plays a role in regulation of PRL gene expression.

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ADENOVIRAL VECTOR MEDIATED IN VIVO GENE TRANSFER OF BDNF PROMOTE FUNCTIONAL RECOVERY AFTER FACIAL NERVE CRUSH INJURY (안면신경 압박손상 후 Adenovirus 매개 BDNF 유전자 전달을 통한 신경손상 회복에 관한 연구)

  • Yang, Byoung-Eun;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.4
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    • pp.308-316
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    • 2006
  • Objectives Despite considerable advances in technique, experience and skill, the precise place of surgery in the treatment of facial nerve injury remains uncertain. We designed a facial nerve crush injury model in rats and evaluated the recovery of crushed nerve which is the most common injury type of facial nerve using adenovirus vector mediated in vivo gene transfer of Brain derived neurotrophic factor(BDNF). Materials and methods In 48 Sprague Dawley rats, we made a facial nerve crush injury model to main trunk before the furcation, and injected a $10^{11}$pfu adenoviral BDNF in experimental group(BDNF adenoviral injection group; ad-BDNF) and $3{\mu}l$ saline in control group(Saline injection group; saline). After a period of regeneration from 10 to 40 days, nerve regeneration was evaluated with functioinal test (vibrissae and ocular movement), electrophysiologic study(threshold, peak voltage, conduction velocity) and histomorphometric study of axon density. Results Vibrissae and ocular movement, threshold and conduction velocity improved as time elapse in both group, however axon density was increased significantly only in experimental group. Functional test in 10 days and 20 days showed no difference between experimental group and control group. Vibrissae movement, threshold, conduction velocity and axon density in 30 days revealed that the regeneration in quality of experimental group was significantly superior to that of control group. Conclusion In general, there is tendency for nerve regeneration in experimental group (BDNF-adenovirus injection group) during 40 days, functional recovery was detected successfully after facial nerve crush in 30 days postoperatively.

Hydroalcoholic Extract of Scrophularia Striata Attenuates Hypertrophic Scar, Suppresses Collagen Synthesis, and Stimulates MMP2 and 9 Gene Expression in Rabbit Ear Model

  • Zarei, Hatam;Tamri, Pari;Asl, Sara Soleimani;Soleimani, Meysam;Moradkhani, Shirin
    • Journal of Pharmacopuncture
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    • v.25 no.3
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    • pp.258-267
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    • 2022
  • Objectives: Hypertrophic scars (HSs) are caused by abnormal wound healing. To date, no standard treatment has been made available for HSs. Scrophularia striata has been reported to accelerate wound healing and has the potential to prevent HS formation. In this study, we investigated the anti-scarring effects of S. striata extract (SSE) in a rabbit ear model of scarring. Methods: In this study, New Zealand white rabbit (weight: 2.3-2.5 kg) were used. In the prevention phase of the study, three test groups received 5%, 10%, and 15% ointments of SSE in the Eucerin base, the fourth group received Eucerin, and the fifth group received no treatment. The samples were obtained on day 35 after wounding. In the treatment phase of the study, the test groups received an intralesional injection of SSE (5%, 10%, and 15%), the fourth group received an intralesional injection of triamcinolone, the fifth group received a solvent (injection vehicle), and the sixth group received no treatment. To evaluate the anti-scarring effects of SSE, the scar elevation index (SEI), epidermis thickness index (ETI), collagen deposition, and MMP2 and MMP9 gene expression were evaluated. Results: A significant reduction in SEI, ETI, and collagen deposition was noted in animals treated with SSE compared with the control groups. In addition, topical SSE stimulated MMP2 and MMP9 gene expression. Conclusion: The findings of this study demonstrate the potential for SSE in the prevention and treatment of HS. SSE could be prepared as an appropriate formulation to treat wounds and prevent abnormal scarring.

Effects of oral administration and herbal acupuncture with Sayeoksanhap-Pyeongweisan-Gamibang(SPG) on gene exdpression in rats damaged by CCl4 (사역산합평위산가미방(四逆散合平胃散加味方)의 경구 투여와 약침이 사염화탄소에 의한 간 손상 모델 백서의 유전자 변화에 미치는 영향)

  • Kim, Hyung-Woo;Jeong, Byeong-Han;Kim, Gye-Yeop;Kim, Young-Kyun;Baek, Jin-Woong;Cho, Su-In
    • The Korea Journal of Herbology
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    • v.22 no.2
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    • pp.163-168
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    • 2007
  • Objectives : This study was designed to investigate effects of Sayeoksanhap-Pyeongweisan-Gambang (SPG) on gene expression in rats damaged by CCl4 Methods : We investigated the effects of SPG on gene expression in terms of microarray methods in rat liver which were obtained from rats damaged by CCl4. Results : Decreased gene expressions, which were induced by single injection of CCl4, were restored to those in normal rats by administration of SPG or herbal acupuncture. In acupuncture group, gene expressions were restored by 80% of those in control group. In oral administration group and combination group, gene expressions were restored above 90% of those in cuntrol group. Conclusion : These results suggest that oral administraion of SPG was useful to protect liver against CCl4 by its restoration of gene expressions in liver resected from rat damaged by CCl4.

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Enhancement of Anti-tumor Activity of Newcastle Disease Virus by the Synergistic Effect of Cytosine Deaminase

  • Lv, Zheng;Zhang, Tian-Yuan;Yin, Jie-Chao;Wang, Hui;Sun, Tian;Chen, Li-Qun;Bai, Fu-Liang;Wu, Wei;Ren, Gui-Ping;Li, De-Shan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7489-7496
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    • 2013
  • This study was conducted to investigate enhancement of anti-tumor effects of the lentogenic Newcastle disease virus Clone30 strain (NDV rClone30) expressing cytosine deaminase (CD) gene against tumor cells and in murine groin tumor-bearing models. Cytotoxic effects of the rClone30-CD/5-FC on the HepG2 cell line were examined by an MTT method. Anti-tumor activity of rClone30-CD/5-FC was examined in H22 tumor-bearing mice. Compared to the rClone30-CD virus treatment alone, NDV rClone30-CD/5-FC at 0.1 and 1 MOIs exerted significant cytotoxic effects (P<0.05) on HepG2 cells. For treatment of H22 tumor-bearing mice, recombinant NDV was injected together with 5-FC given by either intra-tumor injection or tail vein injection. When 5-FC was administered by intra-tumor injection, survival for the rClone30-CD/5-FC-treated mice was 4/6 for 80 days period vs 1/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC and saline alone, respectively. When 5-FC was given by tail vein injection, survival for the rClone30-CD/5-FC-treated mice was 3/6 vs 2/6, 0/6 and 0/6 for the mice treated with rClone30-CD, 5-FC or saline alone, respectively. In this study, NDV was used for the first time to deliver the suicide gene for cancer therapy. Incorporation of the CD gene in the lentogenic NDV genome together with 5-FC significantly enhances cell death of HepG2 tumor cells in vitro, decreases tumor volume and increases survival of H22 tumor-bearing mice in vivo.