An Angiotensin I converting enzyme polymorphism is associated with clinical phenotype when using differentiation-syndrome to categorize korean bronchial asthma patients.

In this study, genetic analysis was conducted to investigate the association of angiotensin I converting enzyme (ACE) gene polymorphism with clinical phenotype based on differentiation-syndrome of bronchial asthma patients. Differentiation-syndrome is a traditional Korean medicine (TKM) theory in which patients are classified into a Deficiency Syndrome Group (DSG) and an Excess Syndrome Group (ESG) according to their symptomatic classification. For this study, 110 participants were evaluated by pulmonary function test. Among them, 39 patients were excluded because they refused genotyping. Of the remaining patients, 52 with DSG of asthma (DSGA) and 29 with ESG of asthma (ESGA), as determined by the differentiation-syndrome techniques were assessed by genetic analysis. ACE insertion/deletion (I/D) polymorphism analysis was conducted using polymerase chain reaction (PCR). Student's t, chi-square, Fisher and Hardy-Weinberg equilibrium tests were used to compare groups. No significant differences in pulmonary function were observed between DSGA and ESGA. The genotypic frequency of ACE I/D polymorphism was found to differ slightly between DSGA and ESGA (P = .0495). However, there were no significant differences in allelic frequency observed between DSGA and ESGA (P = .7006, OR = 1.1223). Interestingly, the allelic (P = .0043, OR = 3.4545) and genotypic (P = .0126) frequencies of the ACE I/D polymorphism in female patients differed significantly between DSGA and ESGA. Taken together, the results presented here indicate that the symptomatic classification of DSGA and ESGA by differentiation-syndrome in Korean asthma patients could be useful in evaluation of the pathogenesis of bronchial asthma.

Effects of Bupleurum falcatum and its combination with an angiotensin II receptor blocker on cytokine and chemokine expression in human mesangial cells.

This study aimed to investigate the inhibitory effect of Bupleurum falcatum and its combination with angiotensin II receptor blocker (ARB) on cytokine and chemokine production in cultured human mesangial cells. Human mesangial cells were isolated and cultured in Dulbecco's modified Eagle's medium culture medium. Bupleurum falcatum, ARB, and the combination of the two were added to human mesangial cells. Cytokine and chemokine levels were analysed using an enzyme-linked immunosorbent assay. There were no significant differences in the expression of IL-1ss, IL-2 or TNF-a between controls and the experimental groups. However, IL-11 and monocyte chemoattractant protein-1 (MCP-1) levels were significantly reduced in response to ARB, Bupleurum falcatum, or their combination when compared with controls. IL-8 expression was reduced significantly only in cells treated with ARB. Both Bupleurum falcatum and ARB treatments alone reduced the cytokine concentration, but there was not a stronger reduction when the two drugs were combined. It was shown that Bupleurum falcatum inhibited cytokine production in human mesangial cells. However, there were no additive effects on the suppression of cytokine production when Bupleurum falcatum was combined with ARB. Further studies are needed to elucidate the renoprotective effects of Bupleurum falcatum.

Analgesic effects by electroacupuncture were decreased in inducible nitric oxide synthase knockout mice.

BACKGROUND: This study was designed to investigate the involvement of inducible nitric oxide synthase (iNOS) in electroacupuncture (EA)-induced analgesia. METHODS: Two and 100 Hz EA stimulation were applied at acupoint ST 36 (Zusanli) in iNOS knockout mice (n = 28). Needles were inserted 5 mm in depth in ST36. After insertion, the needles were fixed in situ with adhesive tape. EA was applied after the basal threshold determination. The EA parameters were set as follows: constant square wave current output (pulse width: 0.6 ms at 2 Hz and 0.2 ms at 100 Hz) and 2 mA intensities. EA was performed for 30 minutes and tail-flick latencies (TFLs) were evaluated every 15 minutes for 1 hour. RESULTS: In 2 Hz EA stimulation, the tail-flick response (TFR) of wild-type mice for durations of 0, 15, 30, 45 and 60 minutes were 2.70 +/- 0.26, 4.19 +/- 0.37, 4.17 +/- 0.34, 3.57 +/- 0.27 and 3.39 +/- 0.32 seconds of TFLs. Meanwhile, iNOS -/- mice showed 4.10 +/- 0.33, 4.77 +/- 0.24, 5.26 +/- 0.30, 4.48 +/- 0.33 and 5.00 +/- 0.41 seconds of TFLs. In 100 Hz EA stimulation, the TFR of wild-type mice were 3.01 +/- 0.24, 4.67 +/- 0.31, 4.76 +/- 0.25, 4.04 +/- 0.45 and 4.26 +/- 0.30 seconds of TFLs. The iNOS -/- mice were 4.33 +/- 0.16, 5.29 +/- 0.28, 5.06 +/- 0.35, 4.52 +/- 0.17 and 4.80 +/- 0.28 seconds of TFLs. Wild-type mice exhibited 63.9% increase in TFL compared to the baseline after 2 Hz EA, whereas the iNOS knockout mice exhibited 32.9% increase in TFL. The TFL after 100 Hz EA showed similar trends: 66.5% increase in TFL in wildtype mice and 18.3% increase in the iNOS knockout mice. CONCLUSION: The present findings suggested that iNOS may play a crucial role in both low- and high-frequency EA-induced analgesic effects.

Peroxisome proliferator-activated receptor-gamma Pro12Ala polymorphism is associated with the susceptibility to ischemic stroke in Taeeumin classified by Sasang medicine.

BACKGROUND: Sasang constitutional medicine classifies mankind into four constitutional types according to individual psychologic and physical traits. We hypothesized that differences among constitutional types might be explained by genetic variations. METHODS: To evaluate the hypothesis, we determined the possible association in ischemic stroke patients (n = 134) of peroxisome proliferator-activated receptor (PPAR)-gamma with four constitutional types of Sasang medicine. The constitutional type of each patient and control subject (n = 129) was classified and genotyped for PPAR-gamma polymorphism Pro12Ala by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) methods. RESULTS: The distribution of the Pro/Ala genotypes in the ischemic stroke patients was not significantly different from that of healthy controls [odds ratio (OR)= 0.46; p = 0.1214]. However, very interestingly, we observed that all six Pro/Ala genotypes in ischemic patients were Taeeumin, one of four constitutional types of Sasang medicine. Statistical analysis revealed that Pro/Ala genotype in Taeeumin increases almost 15-fold the susceptibility to ischemic stroke compared to other constitutional types, Taeyangin, Soyangin or Soeumin (OR= 14.72; p = 0.0110). CONCLUSION: From the results in this study, we might suggest that Pro/Ala genotype in Taeeumin is associated with the susceptibility to ischemic stroke. To the author's best knowledge, this is the first report to study on genetic level the potential relationship between ischemic stroke and Sasang constitutional medicine, one of traditional Korean medicines (TKM). Authors hope that this study could provide a new approach for the study of ischemic stroke and merit further research.

Acupuncture application for neurological disorders.

BACKGROUND: Acupuncture has been widely used for a range of neurological disorders. Despite its popularity, the evidence to support the use of acupuncture is contradictory. METHODS: This review was designed to summarize and to evaluate the available evidence of acupuncture for neurological disorders. RESULTS: Most of the reviewed studies suffer from lack of methodological rigor. Owing to paucity and poor quality of the primary studies, no firm conclusion could be drawn on the use of acupuncture for epilepsy, Alzheimer's disease, Parkinson's disease, ataxic disorders, multiple sclerosis, amyotrophic lateral sclerosis and spinal cord injury. For stroke rehabilitation, the evidence from recent high-quality trials and previous systematic reviews is not convincing. CONCLUSION: More rigorous trials are warranted to establish acupuncture's role in neurological disorders.

fMRI review on brain responses to acupuncture: the limitations and possibilities in traditional Korean acupuncture.

BACKGROUND: Since the late 1990s, imaging studies have allowed the visualization of brain response to acupuncture stimulation with alteration in blood flow. In 1998, the first functional magnetic resonance imaging (fMRI) study for the Korean acupuncture was published and thereafter, many subsequent neuroimaging studies with acupuncture have been published and revealed that acupuncture modulates central nervous system in human being. The purpose of this review is to summarize and evaluate the acupuncture studies using fMRI. METHODS: A survey of computerized literature searches for experimental studies of acupuncture studies using fMRI revealed that a total of 19 studies were published from 1998 to 2005. RESULTS: These studies have mainly focused on the correlation between acupoints and corresponding brain cortices, acupoint-specific patterns of brain activity. The analgesic effect of acupuncture implied that the pain network, including the hypothalamus and limbic system, was induced by different kinds of acupuncture stimulation. There are still several limitations in these experimental designs. CONCLUSION: We reviewed studies that contributed to an understanding of the neurophysiologic mechanisms of acupuncture with the hope that this review will be of benefit to the future traditional Korean acupunctural fMRI studies.

Protein array analysis of cytokine levels on the action of acupuncture in carrageenan-induced inflammation.

BACKGROUND: The aim of this study was to investigate the anti-inflammatory effect of acupuncture on acute paw edema induced by carrageenan (CR) injection and to detect differential cytokine responses in response to acupuncture stimulation using protein array technology. METHODS: Control group was injected with CR (1%, 50 mul) into the plantar surface of the male Sprague-Dawley rats. Acupuncture group was stimulated with acupuncture at Zusanli (ST36) 30 minutes after CR injection. Rat cytokine antibody array coated with 19 specific cytokine antibodies were probed with protein samples and the relative cytokine levels were investigated. RESULTS: Acupuncture stimulation significantly inhibited the inflammatory response to CR injection. Compared to control group, three cytokines, interleukin-6 (IL-6), beta-nerve growth factor (beta-NGF) and tissue inhibitors of metalloproteinase-1 (TIMP-1), showed significantly decreased expression levels in the acupuncture group. The other 16 cytokines did not exhibit significant changes between two groups. CONCLUSION: These results indicate that acupuncture markedly inhibited CR-induced edema and modulated the expressions of certain cytokines in response to CR-induced inflammation. These findings might give us a clue in elucidating the underlying mechanism of anti-inflammatory effect of acupuncture.

Panax ginseng increases hypoxia-induced down-regulated cellular response related genes in human neuroblastoma cells, SK-N-MC.

BACKGROUND: Many studies have suggested that hypoxia plays a crucial role in the pathogenesis of various neurological disorders. To determine protective effect of Panax ginseng (PG) on hypoxia (0.1% O(2))-induced cell death in human neuroblastoma cells SK-N-MC, we profiled the gene expression among hypoxia, PG-treated hypoxia and normoxia groups. METHODS: To determine protective effect on hypoxia-induced cytotoxicity of PG, we performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. We compared the gene expression profiles among hypoxia, PG-treated hypoxia (100 mug/ml, 6 hours) and normoxia groups using 8K human cDNA microarray analysis. Additionally, in order to identify differentially expressed genes between hypoxia and PG-treated hypoxia groups, hierarchical clustering of genes was also performed. RESULTS: MTT assay showed that PG protected hypoxia-induced cell death. In cDNA microarray analysis, hypoxia remarkably down-regulated IGF-II mRNA-binding protein 3 (IMP-3), integrin alpha 2 (ITGA2), syndecan binding protein (SDCBP), insulin-like growth factor binding protein 3 (IGBP3) and M-phase phosphoprotein 10 (MPHOSPH10), belonging to category of cellular physiologic response (global M<-3.5). In cluster analysis, 1428 genes exhibited differential expression levels between hypoxia and PG-treated hypoxia groups. Of them, the expressions of 11 genes were increased more than two-fold by PG treatment compared to those in hypoxia group. Particularly, of 11 genes, the expression levels of cellular physiologic response related genes such as MPHOSPH10, IMP-3 and SDCBP, which markedly down-regulated by hypoxia, are increased more than four-fold by PG treatment, compared to hypoxia group. CONCLUSION: In summary, hypoxia induced down-regulation of cellular physiologic response related genes in human neuroblastoma cells, SK-N-MC, and PG ameliorated the hypoxia-induced down-regulation of such genes. These results indicate possible usage of PG in hypoxia-induced neuronal injury including ischemia, trauma and degenerative diseases.

Suppressive effect by Hizikia fusiforme on the production of tumor necrosis factor in BV2 murine microglial cells.

BACKGROUND: Hizikia fusiforme has been commonly used as food in Korea. Antioxidant effect of Hizikia fusiforme, however, was recently reported. Thus, herein, we investigated the effect of Hizikia fusiforme on the production and expression of tumor necrosis factor (TNF), a major proinflammatory mediator, in lipopolysaccharide (LPS)-activated BV2 microglial cells. METHODS: Cells were pre-treated with 5 or 50 mug/ml Hizikia fusiforme and treated with 1 mug/ml LPS. The production of TNF was measured by enzyme-linked immunosorbent assay (ELISA). The effect of Hizikia fusiforme on the expression of TNF was also performed by immunoblot analysis and reverse transcription-polymerase chain reaction (RT-PCR). Activation of nuclear factor kappab (NFkappab) was determined by electrophoretic mobility shift assay (EMSA). RESULTS: We observed that Hizikia fusiforme decreased the production of TNF. The inhibitory effect of the Hizikia fusiforme on the expression of TNF was confirmed by immunoblot and RT-PCR analyses. In addition, EMSA experiment revealed that Hizikia fusiforme blocked the LPS-induced activation of NFkappab. CONCLUSION: The present study suggests that Hizikia fusiforme may suppress LPS-stimulated TNF production via inhibition of NFkappab in murine microglial cells.

Neuroprotective effect by Dammishimgyu-herbal acupuncture against H(2)O(2)-induced apoptosis in human neuroblastoma, SH-SY5Y cells.

BACKGROUND: The free radical is involved in neuronal cell death in human neurodegenerative disease. Dammishimgyu (DMSG)-herbal acupuncture has been used to treat neurological disorders in Korea. The present study was aimed to investigate the neuroprotective effect of DMSG-herbal acupuncture against H(2)O(2)-induced apoptosis in human neuroblastoma cell line, SH-SY5Y. METHODS: The neuroprotective effect of DMSG-herbal acupuncture on H(2)O(2) induced apoptosis was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, reverse transcription-polymerase chain reaction (RT-PCR), western blots and nitrite assay. RESULTS: In this study, 100 microM H(2)O(2)-treated cells decreased the cell viability with apoptotic features and increased the production of nitric oxide (NO). However, 0.1% DMSG treatment after exposure to 100 microM H(2)O(2) inhibited both H(2)O(2)-stimulated mRNA and protein expressions of BCL2-associated X protein (BAX) and caspase 3 apoptosis-related cysteine peptidase (CASP3). In addition, 0.1% DMSG treatment inhibited the increased NO production induced by H(2)O(2). CONCLUSION: These results suggest that DMSG-herbal acupuncture shows protective effect against H(2)O(2)-induced neuronal damage.

Epidermal growth factor gene polymorphism is different between schizophrenia and lung cancer patients in Korean population.

Low incidence of cancer in schizophrenia is one of the interesting puzzles in psychiatric field over decades. Analysis of genetic difference between schizophrenia and lung cancer might provide us with possible clues to understand molecular mechanisms of pathophysiology of schizophrenia. Epidermal growth factor (EGF), one of the potent growth promoting factors, has been studied for its roles in cancer development. EGF is also known to be involved in cognitive function. In order to analyze the genetic difference between schizophrenia and lung cancer, polymorphism of EGF gene was studied from 174 schizophrenia patients, 122 lung cancer patients and 132 controls in Korean population. Genotype frequency analysis of EGF gene (AluI restriction site, 5'-UTR, rs4444903) in the EGF gene was studied. The genotype and allele frequencies of the AluI polymorphism showed significant differences between schizophrenia and lung cancer patients [p<0.0001; p<0.0001, odds ratio (95% CI), 0.3690 (0.2600-0.5236)]. When compared with controls, schizophrenia patients showed no significant differences from controls in genotype and allele frequencies [p=0.5151; p=0.3516, odds ratio (95% CI), 0.8589 (0.6235-1.1830)]. However, lung cancer patients showed significant differences from controls in genotype and allele frequencies [p<0.0001; p<0.0001, odds ratio (95% CI), 2.3275 (1.6082-3.3687)]. These results indicate that schizophrenia is not associated with AluI polymorphism of EGF gene and EGF gene polymorphism is different between schizophrenia and lung cancer patients.

Estrogen receptor-alpha gene haplotype is associated with primary knee osteoarthritis in Korean population.

Estrogen and estrogen receptors (ERs) are known to play important roles in the pathophysiology of osteoarthritis (OA). To investigate ER-alpha gene polymorphisms for its associations with primary knee OA, we conducted a case-control association study in patients with primary knee OA (n = 151) and healthy individuals (n = 397) in the Korean population. Haplotyping analysis was used to determine the relationship between three polymorphisms in the ER-alpha gene (intron 1 T/C, intron 1 A/G and exon 8 G/A) and primary knee OA. Genotypes of the ER-alpha gene polymorphism were determined by PCR followed by restriction enzyme digestion (PvuII for intron 1 T/C, XbaI for intron 1 A/G, and BtgI for exon 8 G/A polymorphism). There was no significant difference between primary knee OA patients and healthy control individuals in the distribution of any of the genotypes evaluated. However, we found that the allele frequency for the exon 8 G/A BtgI polymorphism (codon 594) was significantly different between primary knee OA patients and control individuals (odds ratio = 1.38, 95% confidence interval = 1.01-1.88; P = 0.044). In haplotype frequency estimation analysis, there was a significant difference between primary knee OA patients and control individuals (degrees of freedom = 7, chi2 = 21.48; P = 0.003). Although the number OA patients studied is small, the present study shows that ER-alpha gene haplotype may be associated with primary knee OA, and genetic variations in the ER-alpha gene may be involved in OA.

Association of estrogen receptor 1 intron 1 C/T polymorphism in Korean vitiligo patients.

BACKGROUND: Vitiligo is a common disease characterized by cutaneous white maculae due to loss of melanocytes. It is a polygenic disease, however, the exact pathogenesis of vitiligo is not yet known. The estrogen receptor (ESR) 1 gene was selected as a candidate gene because some researchers treated vitiligo successfully with the steroid-thyroid hormone mixture containing estrogen. Furthermore ESR was expressed in the melanocytes which have an important role in the pigmentation. The polymorphisms of ESR1 gene in vitiligo patients was not reported yet. OBJECTIVE: To determine whether polymorphisms of ESR1 gene were associated with susceptibility to vitiligo patients in Korean population. METHODS: We conducted case-control association study of vitiligo patients (120) and healthy controls (254). Genotypes of ESR1 gene (intron 1 C/T, exon 4 C/G, and exon 8 A/G) were determined by polymerase chain reaction followed by restriction enzyme digestion. RESULTS: Intron 1 T/C allele frequency was significantly different between patients and controls (P = 0.034). Intron 1 T/C genotype distribution (P = 0.021) and allele frequency (P = 0.013) were different between female vitiligo patients and female controls. Intron 1 T/C allele frequency showed significantly difference between generalized type of vitiligo patients and controls (P = 0.044). Genotype distributions and allele frequencies of exon 4 C/G and exon 8 A/G polymorphisms were not different between patients and controls. CONCLUSION: The present study suggests that ESR1 may be a possible risk factor for female or generalized type of vitiligo patients.

Association between polymorphism in intron 1 of cocaine- and amphetamine-regulated transcript gene with alcoholism, but not with bipolar disorder and schizophrenia in Korean population.

Cocaine- and amphetamine-regulated transcript (CART) peptide is a novel neuropeptide involved in feeding, drug reward, and stress. We hypothesized that the polymorphism of CART gene might be related with susceptibility to neuropsychiatric diseases such as alcoholism, bipolar disorder, and schizophrenia. The polymorphism (rs2239670) in intron 1 was selected for study among other single nucleotide polymorphisms (SNPs) located at the area of CART gene, because it had not been tested until to date. The study included patients of alcoholism (100), bipolar disorder (76) and schizophrenia (169) from the Korean population. Healthy controls for bipolar disorder and schizophrenia consisted of 333 individuals. For alcoholism, both patient group and control subjects included only male. The restriction fragment length polymorphism (RFLP) using the AvaII restriction enzyme was designed to analyze the selected SNP. The distribution of GG, GA, and AA genotypes in the 333 controls was 50.2, 41.1, and 8.7%, respectively. The frequency of G and A alleles in the 333 controls was 70.7 and 29.3%, respectively. The distribution of genotype and allele frequencies of the AvaII polymorphism showed a significant difference between alcoholism and controls (P = 0.037 and P = 0.044). However, the AvaII polymorphism of the CART gene did not show association with bipolar disorder and schizophrenia. In conclusion, we report for the first time that the AvaII polymorphism (rs2239670) in intron 1 of the CART gene is associated with alcoholism in the Korean male population.

Association of IL-1beta, IL-1ra, and TNF-alpha gene polymorphisms in childhood nephrotic syndrome.

We investigated the association between IL-1beta, IL-1ra, and TNF-alpha gene polymorphisms and childhood nephrotic syndrome (NS). We analyzed the genetic polymorphism of IL-1beta, IL-1ra, and TNF-alpha genes in 152 patients with childhood NS and 292 healthy adult controls. The C to T exchange at position -511 of IL-1beta and the G to A at -308 of the TNF-alpha gene were genotyped. Five alleles of the IL-1ra gene were identified and designated as IL1RN*1, IL1RN*2, IL1RN*3, IL1RN*4, and IL1RN*5, according to the variable number of tandem repeats in intron 2. The allele frequencies of IL-1beta1 (-511C), IL-1beta2 (-511T), TNF1 (-308G), and TNF2 (-308A) were 53.0, 47.0, 92.1, and 7.9%, respectively, in the childhood NS group. This was not significantly different from normal controls. In the childhood NS group, the allele frequencies of IL1RN*1, IL1RN*2, IL1RN*3, IL1RN*4, and IL1RN*5 were 90.8, 7.6, 1.6, 0, and 0% [IL1RN*1 odds ratio (OR)=0.296, P=0.0001, IL1RN*2 OR=3.902, P=0.0002]. A high allele frequency of IL1RN*2 and a lower allele frequency of IL1RN*1 were found in childhood NS, although there was no association with IL-1beta and TNF-alpha. A high allele frequency of the IL1RN*2 allele may affect disease susceptibility in childhood NS.

Association of angiotensin converting enzyme gene I/D polymorphism of vitiligo in Korean population.

Vitiligo (leukoderma) is an acquired idiopathic hypomelanotic disorder characterized by the circumscribed depigmented patches. Vitiligo is a polygenic disease. The exact pathogenesis is not yet known. The angiotensin converting enzyme (ACE) gene was selected as a candidate gene as ACE plays an important role in the physiology of the vasculature, blood pressure and inflammation, and its relationship with various diseases, including autoimmune diseases, has been widely investigated. The I/D polymorphism of ACE gene in vitiligo patients has not been reported. In this study, we investigated ACE gene polymorphism in 120 vitiligo patients and in 429 healthy volunteers in Korea. The ACE gene genotype distribution (P = 0.032) and allele frequency (P = 0.012) were significantly different between vitiligo patients and healthy controls. This study suggests that the ACE gene polymorphism has a strong association with the development of vitiligo in Korean patients.

Acupuncture increases neuropeptide Y expression in hippocampus of maternally-separated rats.

Maternal separation (MS) is a risk factor in the development of mood-related disorders such as depression. Human and animal studies support the involvement of neuropeptide Y (NPY) in the pathology of depression. To investigate the effect of acupuncture on depression-like behavior and examine changes in NPY expression associated with MS, we observed body weight and locomotor activity, and performed NPY immunohistochemistry in the hippocampus. MS for 7 days beginning on postnatal day 14 induced a significant decrease in body weight and locomotion, while acupuncture treatment at acupoint Shenmen (HT7) resulted in a significant increase in both. NPY-immunoreactive cells in area CA1 and the dentate gyrus were decreased in the MS group, but significantly increased in the acupuncture group. These findings suggest that acupuncture has an effect on the depression-like disorder caused by MS, possibly by modulating NPY expression in the hippocampus.