• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.139-144
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• 2003

A chitinase cDNA named CHT1 was cloned from the hard tick, Haemaphysalis longicornis, and the enzymatic properties of its recombinant proteins were characterized. The CHT1 cDNA encodes 930 amino-acid (aa) residues including a 22 aa putative signal peptide, with the calculated molecular mass of the putative mature protein 104 kDa. The E coli-expressed rCHT1 exhibited weak chitinolytic activity against $4MU-(GlcNAc)_3$. The rCHT1 protein with higher activity was obtained using recombinant Autographa californica multiple nuclear polyhedrosis virus (AcMNPV), which expresses rCHT1 under polyhedrin promoter. These findings suggest that the rCHT1 expressed in baculovirus-mediated Sf 9 cells has a high activity than E coli-expressed rCHT1.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.6
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• pp.1127-1133
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• 2002

Cheonghunhwadam-tang(CHT) have been used in oriental medicine for many centuries as a therapeutic agent of vertigo by wind, fire and phlegm. CHTGR was CHT adding Gastrodae Rhizoma. The effects of CHTGR on the regional cerebral blood flow(rCBF) and mean arterial blood pressure(MABP) is not known. A comparative Study of action-mechanism of CHT and CHTGR on the cerebral hemodynamics is not known too. Therefore, purpose of this Study was to investigate effects of CHT and CHTGR on the rCBF and MABP, compare action-mechanism of CHT and CHTGR on the rCBF and MABP. The changes of rCBF and BP was determinated by Laser-Doppler Flowmetry(LDF). The results were as follows ; CHT extract was increased rCBF in a dose-dependent, but was not changed MABP compared with CHT non-treated group. CHTGR extract was decreased rCBF and MABP compared with CHTGR non-treated group in a dose-dependent. Action of CHT is not related with adrenergic β-receptor, cyclooxygenase and guanylate cyclase, but action of CHTGR is related with guanylate cyclase.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.6
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• pp.1287-1291
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• 2009

The genes of cyanide hydratase(CHT), a kind of nitrilases whichhydrolyze cyanide to formamide were extracted from N. crassa and A. nidulans, the two fungal strains. The recombinant forms of the CHT originated from N. crassa and A. nidulans were prepared with N-terminal hexahistidine purificationtags or no tags, and expressed in E. coli. The enzymes were purified using immobilized metal affinity chromatography. They were compared according to their pH activity profiles, and kinetic parameters. The N. crassa CHT has the wider pH range of activity above 50% and three-fold higher turnover rate (6.6 ${\times}$ $10^8$ $min^{-1}$) than the A. nidulans, meanwhile the CHT of A. nidulans has the higher $K_m$ value. Expression of CHT in both N. crassa and A. nidulans were induced by the presence of KCN, regardless of any presence of nitrogen sources. Max. 82% of KCN was degraded in 60 min for biological degradation tests.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.4
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• pp.85-105
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• 2005

Purpose : Investigate the effects of Chungganhaewooltang(CHT) on immobilization-stress or cold-stress in C576BL/6J mice. Methods : Male C57BL/6J 30 mice of weighting 18${\pm}$2g, were divided into sixs groups including the immobilization-stress group(5heads), after immobilization-stress CHT oral administration(500mg/kg) groups(5heads), cold-stress group(5heads) and after cold-stress CHT oral administration(500mg/kg) groups(5heads). then we observed changes in the serum histamine and corticosterone level and changes immune system Results : Immobilization-stress or cold-stress increased the serum level of histamine and corticosterone. CHT decreased the serum level of histamine and corticosterone increased by cold-stress. CHT inhibited the release of histamine from mast cells at the concentration of 0.1 mg/ml. In addition, immobilization-stress or cold-stress decreased the cell viability of murine thymocytes and splenocytes. CHT increased the cell viability of thymocytes decreased by immobilization-stress or cold-stress, but did not affect the cell viability of splenocytes decreased by immobilization-stress or cold-stress. Also immobilization-stress or cold-stress increased DNA fragmentation of thymocytes and splenocytes. CHT decreased DNA fragmentation of thymocytes increased by immobilization-stress or cold-stress, but did not affect DNA fragmentation of splenocytes increased by immobilization-stress or cold-stress. Immobilization-stress increased the population of thymic $CD4^+$ cells. CHT decreased the population of thymic $CD4^+$ cells increased by immobolization-stress. Immobilization-stress or cold-stress decreased the population of $B220^+$ cells and increased the population of $thy1^+$ cells. CHT decreased the population of $thy1^+$ cells increased by immobilization-stress or cold-stress. Immobilization-stress or cold-stress increased the population of splenic $CD4^+$ cells and $CD8^+$ cells. CHT decreased the population of splenic $CD4^+$ cells increased by immobolization-stress or cold-stress. Immobilization-stress or cold-stress decreased the production of ${\gamma}-interferon$(IFN) interleukin(IL)-2 and IL-4. CHT enhanced the production of ${\gamma}-IFN$ decreased by immobilization-stress or cold-stress but did not affect the production of IL-2 and IL-4 decreased by immobilization-stress or cold-stress. Furthermore, Immobilization- stress or cold-stress decreased the phagocytic activity of peritoneal macrophages and the production of nitric oxide. CHT enhanced the phagocytic activity and nitric oxide production decreased by cold-stress. Conclusion : CHT may be useful for the prevention and treatment of stress via suppression of serum histamine and corticosterone level and enhancement of immune response.

• Journal of Haehwa Medicine
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• v.20 no.2
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• pp.29-39
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• 2012

To investigate the clinical aspects of CHT in atopic dermatitis (AD) treatments, the effect of CHT in anti-oxidative and anti-inflammatory cytokines were tested. 100% or higher cell viability was observed in all tested groups from 25 to 200 ${\mu}g/m{\ell}$ using Raw 264.7 cells. CHT showed dose-dependent DPPH scavenging activity, with more than 90% scavenging activities at 800 ${\mu}g/m{\ell}$ concentrations. CHT showed dose-dependent suppression activity of ROS production, especially at 200 ${\mu}g/m{\ell}$ of 37.5%. CHT decreased NO production activity, with significant decrease of 33.2% at 200 ${\mu}g/m{\ell}$. IL-6, MCP-1, TNF-${\alpha}$ production rate were decreased by approximately 25% when Raw 264.7 cells were treated with LPS and with CHT of 200 ${\mu}g/m{\ell}$. Also, IL-$1{\beta}$ production rate was decreased by 25% at 100 ${\mu}g/m{\ell}$. The results above indicate that CHT significantly reduces the effect of oxidative and inflammatory cytokines. The use of CHT in dermatitis can be widely suggested.

• The Journal of Internal Korean Medicine
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• v.27 no.2
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• pp.305-315
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• 2006

Objectuves : The purpose of this study was to investigate the effects of Cheunghyeoltang(CHT) and Gami-Cheunghyeoltang(GCHT) on the obesity in rats. Methods : After we divided rats into four different groups, we Provided to normal group general diet and distilled water, to control group high fat diet and distilled water, to CHT group high fat diet and CHT and to GCHT group with high fat diet and GCHT for 9 weeks. Afterward we check the change of body weight, the concentration of AST, ALT, total cholesterol, HDL-cholesterol, LDL-cholesterol, the Atherogenic index and the gene expression of leptin in rats. Results : The increase of body weight were suppressed significantly in the CHT group and GCHT group in comparision with control group, the serum ALT showed significant decrease in the GCHT group in comparison with control group, the serum total cholesterol showed significant decrease in the CHT and GCHT group in comparison with control group, the atherogenic index(AI) showed significant decrease in the GCHT group in comparison with control group and the serum leptin showed increase of gone expression in the GCHT group in comparison with control group. Conclusions : According to the experimental results, it is suggested that CHT is effective to the suppression of body weight's increase, the improvement of hyperlipidemia and the activation of leptin's gene expression, GCHT is effective to the suppression of body weight's increase, the improvement of hyperlipidemia, the decrease of arteriosclerosis and the activation of leptin's gene expression. Therefore CHT and GCHT seem to be applicable to obesity, arteriosclerosis and hyperlipidemia in clinical practice.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1399-1404
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• 2005

In oriental medicine, Cheongpyehwadam-tang (CHT) has long been used for the cure of inflammatory diseases in the lung and bronchus such as bronchitis, bronchial asthma, pneumonia and tuberculosis. It's use is currently further extended for the treatment of allergic asthma. To investigate the anti-inflammatory effects of CHT, we investigated the effects of CHT on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) production, and on the level of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines expression in murine macrophage RAW 264.7 cells. CHT alone did not affect NO or pro-inflammatory cytokines production. In contrast, CHT inhibited LPS-induced NO and proinflammatory cytokines and the levels of LPS-induced iNOS and proinflarnmatory cytokine mRNA in a dose-dependent manner. CHT also inhibited the nuclear factor-kappa B (NF-kB) activation. Taken together, these results suggested that CHT inhibits the production of NO and pro-inflammatory cytokines in RAW 264.7 cells through blockade of NF-kB activation.

• Journal of Neurogastroenterology and Motility
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• v.24 no.4
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• pp.643-655
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• 2018

Background/Aims Irritable bowel syndrome (IBS) is a common disease characterized by intestinal dysmotility, the mechanism of which remains elusive. We aim to determine whether the high-affinity choline transporter 1 (CHT1), a determinant of cholinergic signaling capacity, modulates intestinal motility associated with stress-induced IBS. Methods A rat IBS model was established using chronic water avoidance stress (WAS). Colonic pathological alterations were evaluated histologically and intestinal motility was assessed by intestinal transit time and fecal water content (FWC). Visceral sensitivity was determined by visceromotor response to colorectal distension. RT-PCR, western blotting, and immunostaining were performed to identify colonic CHT1 expression. Contractility of colonic muscle strips was measured using isometric transducers. enzyme-linked immunosorbent assay was used to measure acetylcholine (ACh). We examined the effects of MKC-231, a choline uptake enhancer, on colonic motility. Results After 10 days of WAS, intestinal transit time was decreased and fecal water content increased. Visceromotor response magnitude in WAS rats in response to colorectal distension was significantly enhanced. Protein and mRNA CHT1 levels in the colon were markedly elevated after WAS. The density of CHT1-positive intramuscular interstitial cells of Cajal and myenteric plexus neurons in WAS rats was higher than in controls. Ammonium pyrrolidine dithiocarbamate partly reversed CHT1 upregulation and alleviated colonic hypermotility in WAS rats. Pharmacological enhancement of CHT1 activity by MKC-231 enhanced colonic motility in control rats via upregulation of CHT1 and elevation of ACh production. Conclusion Upregulation of CHT1 in intramuscular interstitial cells of Cajal and myenteric plexus neurons is implicated in chronic stress-induced colonic hypermotility by modulation of ACh synthesis via nuclear factor-kappa B signaling.

• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.145-150
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• 2003

To determine effectively the chitinolytic activity of rCHT1 from the hard tick H. longicornis expressed in baculovirus-mediated Spodoptera frugtperda (Sf) 9 cells, a simple and sensitive assay system was established in solid phase using agarose gel containing ethylene glycol chitin as substrate. The various factors affecting the efficacy of the assay were also investigated. The effects of various temperature, dosages of proteins, pH of media and time courses of reaction were examined to verify the sensitivity of assay for chitinolytic activity of rCHT1 protein. It was found that the optimal reactive conditions were $37^{\circ}C$ of temperature, 12 to 15 hours of reactive times, $0.1{\mu}g$ of protein concentration and pH 5 to 7 of media. Using the assay system designed, the functional activities of H. longicornis rCHT1l protein could be evaluated simply and sensitively.

• The Journal of Korean Medicine
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• v.29 no.3
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• pp.76-87
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• 2008

Objectives: In this study, the author investigated whether Haengso-tang (HST) and Chwiyeon-tang (CHT) affect both in vitro mucin secretion and MUC5AC gene expression in airway epithelial cells and in vivo mucin secretion from animal model for airway mucus hypersecretion. Materials and Methods: Confluent HTSE cells (non-labeled) were chased for 30 min in the presence of HST and CHT to assess the effects of the agents on mucin secretion by enzyme-linked immunosorbent assay (ELISA), with removal of oriental herbal medicine extract from each agent-treated sample by centrifuge microfilter. Also, the effects of the agents on TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. The author also induced hypersecretion of airway mucus by exposure of rats to SO2 for 3 weeks. Effects of orally-administered HST and CHT during 1 week on in vivo mucin secretion from tracheal goblet cells of rats were assessed using ELISA. Results: (1) HST significantly decreased in vitro mucin secretion from cultured HTSE cells. However, CHT did not affect in vitro mucin secretion from HTSE cells; (2) CHT significantly inhibited the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells. However, HST did not affect the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells; (3) CHT significantly inhibited hypersecretion of in vivo mucin. However, HST did not affect hypersecretion of in vivo mucin. Conclusion: These results suggest that CHT can not only affect the secretion of mucin but also the expression of the mucin gene and could be helpful for treating pulmonary disease caused by secretion of mucin.