• Journal of Life Science
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• v.19 no.11
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• pp.1547-1552
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• 2009

This study was carried out to evaluate the supplementation effect of dietary natural plant extracts (NP: $Coxynil^{(R)}$, $Growell^{(R)}$ and $Respowell^{(R)}$) on broiler chickens. Forty thousand male broilers with 7 days adaptation after hatching were fed experiment diets for 34 days. The supplementation effects of NP on growth performance, blood parameters and biopsy were examined with twenty thousand broilers as the treatment group. Twenty thousand broilers for the control group (CON) were fed the diet with salinomycin-6, clopidol-25, enramycin-1, and BMD-2.5. In the diet of the treatment group, the antibiotics were replaced with 0.03%, 0.035% and 0.03% of $Coxynil^{(R)}$, $Growell^{(R)}$ and $Respowell^{(R)}$, respectively. The weight gain of the treatment group was increased but the feed intake was decreased, indicating that feed efficiency was increased compared to the CON. The mortality of the NP group was also lower compared to the CON group (1,008 birds to 1,693 birds), showing positive dietary effects from natural plant extracts. In the activity of infectious bursal disease virus (IBDV) and new cattle disease virus (NDV) antibodies, the NP showed lower antibody titer levels for both of IBDV and NDV compared to the CON. The levels of total cholesterol, HDL-cholesterol, globulin, and IgG in blood did not show significant differences between the groups. In the microscopic tissue analysis, no significant differences were detected. These results may suggest that a complex of three natural plant extracts can be used as alternative antibiotics in broilers.

• The Journal of Korean Medicine
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• v.17 no.1 s.31
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• pp.190-211
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• 1996

Generally the Myasthenia Gravis is classified into two of hereditary factor and acquement. Aquired Myasthenia Gravis was Quite well known to be caused by the autoimmune mechanism. Not in accurateness, on the hereditary, acetylcholine receptor antibody was to be analyzed very high in their parents and brothers. Also Myasthenia Gravis is a chronic disease characterized by voluntary muscle weakness and fatigue. above all, ocular Myasthenia Gravis is characterized clinically by blepharoptosis and external ophthalmoplegia and to be showed abut 90% cases and so oriental medicine can not but deal with myasthenia gravis at blepharoptosis. Accordinglv 20 out patients with Myasthenia Gravis were clinical study and observation as to the sex, age, progress state of MG, blood type, history, main symptom, liking for warm and cool food and tepidity, state of pulse, treatment of acupuncture and administration of oriental medicine etc. The results were as follows. 1. There was investigaed on the frequency of attack for sex, age, oculus dexter, oculus sinister, oculus uterque. Among the 20 patients, the number of female were 60% with 12 cases and male were 40% with 8 cases, therefore it was the rate of 6 : 4. the patients under 10 ages and 40 ages were 20% with 4 cases, 10 ages and 50 ages were 15% with 3 cases, 20, 30, 60 ages were 10% with 2 cases. And then oculus uterque was 90% with 18 cases, oculus sinister was 10% with 2 cases and oculus dexter were none of them. 2. Stage I were 50% with 10 patients, stage $II_A$ were 30% with 6 patients and stage $II_B$ were 20% with 4 patients, on the clinical stage and too class I were 20% with 4, class II were 45% with 9, class III were 35% with 7, in the functional activity the patients with chest heavy were 15% with 3 and hyperthyroidism were 10% with 2. 3. Hospital which patients had used to before came to this hospital were 10 university hospital and 6 local clinic. 4. The duration of disease was from 3 months to 30 years, the patients suffering between 3 months and within 1 year were 25% with 5 cases, 1-2 years were 30% with 6 cases, therefore within 2 years were 55%. 4-5 years were 15%, over 7 years were appeared less than 10%. 5. In the main symptom, all of patients were appeared to be heavy in opening their eyes the patients with blephroptosis were 70% with 14 cases on the oculus uterque, oculus sinisterf and oculus dexter, there were 20% with 4 eases each other in the oculogyation incomplete. visual failing, ophthalmoxerosis, strabismus etc and indigetion, frequency of urine(feel hurt), mild stools(or diarrea), oversensitiveness etc. but in addition, all of the other were 10%. 6. In the distribution of blood type, 0 types were 45% with 9 cases, A types were 25% with 5 cases, B and AB types were 15% each other. 7. For the rates of patients of liking for warm and cool food or tepidity, patients of liking for warm food possess 45% with 9 cases, and cool food possess 35% with 7 and tepidity possess 20% with 4, and then most of patients liking for warm food were females and cool food were much more males than females. 8. Hyunsae(弦細) were 40% with 8 cases, Buhurl(浮滑) were 20% with 4 cases, Hyunsak(弦數) were 15% with 3 cases, and in addition, the others were 10%, among 7 types of pulses. 9. The patients with less than 1 week were 40% with 8 cases, and there were female most of them and over 4 weeks were 20% and 1-2weeks were 15%, in the duration acupuncture treatment. 10. 15 kinds of prescriptions were administrated with oriental medicine from 1 week to 20weeks 1_2 weeks were 25.71% with 9 cases, 3 weeks were 17.14% with 6 cases and 6 weeks were 11.42% with 4 cases and also Gamibaetaugunbitang(加味培土健脾湯) were 28.57% with 10 cases, Gamijeounyongtang(加味正容湯) were 14.28% with 5 cases, Gamibojoongyigitung (加味補中益氣湯), Gamiyinsamyangyoungtung (加味人蔘養榮湯) were 8.57% with 3 cases each other and also Gamisamgitung (加味蔘?湯), Gamisamuloajatung(加味四物五子湯) Gamigoudungum (加味鉤藤飮), etc were applied.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.30-37
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• 2007

It is well-known that Korean mistletoe (Viscum album) extract has an immune activity and anticancer effect. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to examine whether this extract might activate human peripheral monocyte to produce tumor necrosis $factor-\alpha$ $(TNF-\alpha)$. To examine the effect of M11C on the production of $TNF-\alpha$ from monocyte, the monocyte were stimulated by the M11C, and then collected the supernatant (M11C stimulated monocyte-conditioned media; MCM). MCM was treated to the $TNF-\alpha$ sensitive L929 cells, and then L929 cytotoxicity was measured by means of MTT. MCM had cytotoxic effect on L929. And the cytotoxic effect of MCM on L929 was almost abolished by $anti-TNF-\alpha$ antibody. These data indicated that MCM contained $TNF-\alpha$, suggesting the $TNF-\alpha$ generation from M11C-stimulated monocyte. This suggestion was confirmed from the data that $TNF-\alpha$ was highly detected in MCM by immunoblotting technique. M11C effect on $TNF-\alpha$ production from monocyte was in the dose and stimulating time dependent manners. Also the effect of M11C on the expression of $TNF-\alpha$ mRNA from monocyte was shown in the dose and stimulating time dependent manners. As a result, Korean mistletoe extract, M11C, could be used for an immunostimulator.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Journal of Korean Medicine Rehabilitation
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• v.18 no.4
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• pp.39-61
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• 2008

Objectives : The present study was performed to investigate whether acupuncture stimulation in the rats affected regeneration properties of the injured sciatic nerve. A differential effect of acupuncture stimulation on the one point near the spinal nerve root controlling sciatic nerve activity and the other point in the peripheral area subordinated by injured nerve was compared. Materials and Methods: Rat sciatic nerves were injured by crush, and the effects on axonal regeneration on injured sciatic nerves were evaluated by acupuncture stimulation at two different regions. In proximal acupuncture stimulation group, acupuncture stimulation was performed on Huatuo Jiaji(EX B2) points located from L5 to S1 vertebral levels to stimulate the nearest spinal nerve root that innervates sciatic nerves. In distal acupuncture stimulation group, acupuncture stimulation was performed on Zusanli(ST 36) and Weizhong(BL 40) points to stimulate at peripheral area dominated by injured sciatic nerves. Acupuncture stimulation was given every other days for 1 or 2 weeks. Sciatic nerve tissues collected from acupuncture stimulation experimental groups, injury control group, and intact animal group were used for protein analysis by Western blotting or Hoechst nuclear staining. To determine axonal regeneration, Dil fluorescence dye was injected into the sciatic nerve 0.5 cm distal to the injury site in individual animal groups and Dil-labeled cells by retrograde tracing were measured in the DRG at lumbar 5 or in the spinal cord. DRG sensory neurons prepared from individual animal groups were used to measure the extent of neurite outgrowth and for immunofluorescence staining with anti-GAP-43 antibody. Results : Animal groups given proximal or distal acupuncture stimulation showed upregulation of GAP-43 and Cdc2 protein levels in the sciatic nerve at 7 days after injury. Cdk2 protein levels were strongly induced by nerve injury, but did not show changes by acupuncture stimulation. Phospho-Erk1/2 protein levels were elevated by acupuncture stimulation above those present in the injury control animals. These increase in regeneration-associated protein levels appeared to be related with increase cell proliferation in the injured sciatic nerves. Hoechst 33258 staining of sciatic nerve tissue to visualize nuclei of individual cells showed increased Schwann cell number in the distal portion of the injured nerve 7 and 14 days after injury and further increases by acupuncture stimulation particularly at the proximal position. Measurement of axonal regeneration by retrograde tracing showed significantly increased Dil-labeled cells in proximal acupuncture stimulation group compared to distal acupuncture stimulation group and injury control group. Finally, an evaluation of axonal regeneration by retrograde tracing showed increased number of Dil labeled cells in the DRG at lumbar 5 or in the ventral horn of the spinal cord at lower thoracic level at 7 days after nerve injury. Conclusions : The present data show that the proximal acupuncture stimulation at Huatuo Jiaji(EX B2) points governing injured sciatic nerves was more effective for axonal regeneration than the distal acupuncture stimulation. Further studies on functional recovery or associated molecular mechanisms should be critical for developing animal models and clinical applications.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.2 no.2
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• pp.169-177
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• 1999

Purpose: To evaluate the efficacy of interferon alpha therapy with or without prednisolone in children with chronic hepatitis B. Methods: Twenty-eight children (22 boys, 6 girls, mean age 130 months) had seropositive results for HBsAg, HBeAg and HBV DNA; 11 had chronic persistent hepatitis and 17 had chronic active hepatitis. The patients were divided into two groups depending upon their inflammatory activity on liver biopsy, pretreatment serum ALT levels and HBV DNA levels. Fourteen children (group 1: chronic active hepatitis, ALT ${\geq}$ 100 IU/L and HBV DNA ${\leq}$ 100 pg/$300\;{\mu}L$) received interferon alpha 2a 5 $MU/m^2$ of body surface three times weekly for 6 months. Fourteen children (group 2: chronic persistent hepatitis or chronic active hepatitis with ALT < 100 IU/L or HBV DNA > 100 pg/$300\;{\mu}L$) received prednisolone in decreasing daily doses of 60 mg/$m^2$, 40 mg/$m^2$, and 20 mg/$m^2$, each for 2 weeks, followed after 2 weeks by interferon alpha 2a on the same schedule. At the end of therapy, 3 end points were analyzed: HBeAg seroconversion, serum ALT normalization rate and clearance of serum HBV DNA. Results: At the end of treatment, HBe antigen-to antibody seroconversion was higher but not more significant in group 1 than group 2 (71.4% vs. 50.0%). Only one patient in group 2 who lost HBeAg, also cleared HBsAg. ALT normalization was similar in both groups (64.3% in group 1 vs. 55.6% in group 2). Clearance of serum HBV DNA was observed in 78.6% of patients in group 1 and 64.3% in group 2, but no significant differences. Complete response was similarly achieved in both groups (57.1% in group 1 vs. 50.0% in group 2). Interferon alpha therapy with prednisolone priming was well tolerated and all children finished therapy. Conclusion: The combined therapy with prednisolone followed by interferon alpha may be safe and effective in inducing a serological and biochemical remission of the disease in approximately 50% of children with chronic hepatitis B and with a high level of viral replication and less active disease. However, a controlled study should be performed to confirm these results.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.339-353
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• 1994

Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.