Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn2+ were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.
Amino Acid Sequence ; Animals ; Clonorchis sinensis/*enzymology ; Cluster Analysis ; Conserved Sequence ; DNA, Complementary/genetics ; Energy Metabolism ; Gene Expression Profiling ; Mitochondria/metabolism ; Models, Molecular ; Molecular Weight ; Phylogeny ; Protein Conformation ; Sequence Homology, Amino Acid ; Ubiquitin-Protein Ligases/chemistry/*genetics/*metabolism

Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn2+ were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.
Amino Acid Sequence ; Animals ; Clonorchis sinensis/*enzymology ; Cluster Analysis ; Conserved Sequence ; DNA, Complementary/genetics ; Energy Metabolism ; Gene Expression Profiling ; Mitochondria/metabolism ; Models, Molecular ; Molecular Weight ; Phylogeny ; Protein Conformation ; Sequence Homology, Amino Acid ; Ubiquitin-Protein Ligases/chemistry/*genetics/*metabolism

OBJECTIVE: To investigate the frequency, type and severity of cardiac abnormalities in the patients with ankylosing spondylitis and undifferentiated spondyloarthopathy. METHODS: A history, clinical examination, standard 12 lead electrocardiography, two dimensional, M mode, and Doppler echocardiographies were performed on 19 patients with ankylosing spondylitis, 15 patients with undifferentiated spondyloarthropathy and 21 normal controls. RESULTS: 1) Cardiac abnormalities were detected in 8 patients(42.1%) with ankylosing spondylitis. 2) Cardiac abnormalities were detected in 8 patients(53.3%) with undifferentiated spondyloarthropathy including 2 patients with aortic valve abnormalities(mild aortic insufficiency, aortic valve thickening. 3) Cardiac abnormalities were detected in one (4. 8%) among normal controls (mild tricuspid regurgitation). 4) There were sinus bradycardias on electrocardiography in 2 patients among patients witn anl;ylosing spond!litis and in 1 patient among undifferentiated spondyloarthropathy. But there was no conduction disturbance in both groups. 5) The frequency of cardiac abnormality was higher in patients with ankylosing spondylitis and undifferentiated spondyloarthropathy than in normal controls. 6) The mean age, mean disease duration, presence of uveitis, peripheral arthritis, HLA-B27, enthesopathy, Schober test and chest expansion in the patients with ankylosing spondylitis and undifferentiated spondyloarthropathy with cardiac abnormalities were not different from those in the patients without cardiac abnormalities. CONCLUSION: The frequency of cardiac abnormality was higher in patients with ankylosing spondylitis and undifferentiated spondyloarthropathy than in normal controls. The frequency, type and severity of cardiac involvement in patients with ankylosing spondylitis were not different from those in patients with undifferentiated spondyloarthropathy.
Aortic Valve ; Aortic Valve Insufficiency ; Arthritis ; Bradycardia ; Electrocardiography ; HLA-B27 Antigen ; Humans ; Rheumatic Diseases ; Spondylarthropathies* ; Spondylitis, Ankylosing* ; Thorax ; Uveitis

OBJECTIVE: To introduce a newly designed arm sling and to compare the effects on a shoulder subluxation. METHOD: Sixteen hemiplegic patients with shoulder subluxation were evaluated by a simple X-ray with and without the slings and the vertical and horizontal distances on the plain AP views were measured. The newly designed arm sling was compared in terms of the effects of correction with a pouch sling and a Bobath sling. The arm sling designed for this study was developed for the purpose of maintaining patients' hands in a functional position and performing ROM exercise of the shoulder easily. The mean values of the vertical and horizontal distance were compared to determine if there was significant difference of function between the new sling and the conventional slings. Variables such as comfort, choice, and easiness for donning and doffing of the arm slings were evaluated by questions. RESULTS: The new sling provided the patients with good vertical correction of the subluxation (p<0.05) but did not increase the horizontal distance significantly. According to the responses to the questions, the new sling was more comfortable than the conventional slings. CONCLUSION: These results support the effectiveness and the comfort of a new sling to decrease subluxation in hemiplegia. Further study on the long term effects or complication of the new sling is recommended.
Arm ; Hand ; Hemiplegia ; Humans ; Pilot Projects* ; Shoulder*

Objective: : This study aimed to validate the Korean version of the Repeatable Battery for the Assessment of Neuropsychological Status Update (K-RBANS). Methods: : We performed a retrospective analysis of 283 psychiatric and neurosurgery patients. To investigate the convergent validity of the K-RBANS, correlation analyses were performed for other intelligence and neuropsychological test results. Confirmatory factor analysis was used to test a series of alternative plausible models of the K-RBANS. To analyze the various capabilities of the K-RBANS, we compared the area under the receiver operating characteristic (ROC) curves (AUC). Results: : Significant correlations were observed, confirming the convergent validity of the K-RBANS among the Total Scale Index (TSI) and indices of the K-RBANS and indices of intelligence (r=0.47–0.81; p<0.001) and other neuropsychological tests at moderate and above significance (r=0.41–0.63; p<0.001). Additionally, the results testing the construct validity of the K-RBANS showed that the second-order factor structure model (model 2, similar to an original factor structure of RBANS), which includes a first-order factor comprising five index scores (immediate memory, visuospatial capacity, language, attention, delayed memory) and one higher-order factor (TSI), was statistically acceptable. The comparative fit index (CFI) (CFI, 0.949) values and the goodness of fit index (GFI) (GFI, 0.942) values higher than 0.90 indicated an excellent fit. The root mean squared error of approximation (RMSEA) (RMSEA, 0.082) was considered an acceptable fit. Additionally, the factor structure of model 2 was found to be better and more valid than the other model in χ2 values (Δχ2=7.69, p<0.05). In the ROC analysis, the AUCs of the TSI and five indices were 0.716–0.837, and the AUC of TSI (AUC, 0.837; 95% confidence interval, 0.760–0.896) was higher than the AUCs of the other indices. The sensitivity and specificity of TSI were 77.66% and 78.12%, respectively. Conclusion : The overall results of this study suggest that the K-RBANS may be used as a valid tool for the brief screening of neuropsychological patients in Korea.

Background Oenanthe javanica is an aquatic perennial herb originated from East Asia.Nowadays,the effects of Oenanthe javanica have been proven in various disease models.Studies regarding the antioxidant effect of Oenanthe javanica in the kidney are still unclear.Methods This study was therefore performed to investigate the effect of the Oenanthe javanica extract (OJE) in the rat kidney using immunohistochemistry for antioxidant enzymes,copper,zinc-superoxide dismutase (SOD1),manganese superoxide dismutase (SOD2),catalase (CAT) and glutathione peroxidase (GPx).Sprague-Dawley rats were randomly assigned to three groups:(1) normal diet fed-group (normal-group),(2) diet containing ascorbic acid (AA)-fed group (AA-group) as a positive control,(3) diet containing OJE-fed group (OJE-group).AA and OJE were supplied during 28 days.Results The side-effects were not observed in all the groups.Immunoreactivities of SOD1,SOD2,CAT and GPx were easily detected in the distal tubules of the kidney,and their immunoreactivities in the AA-and OJE-groups were increased to about 1.4-1.5 times and 2 times,respectively,compared with those in the normal-group.Conclusion OJE significantly increased expressions of SOD1 & 2,CAT and GPx immunoreactivities in the distal tubules of the rat kidney,and this finding suggests that significant enhancements of endogenous enzymatic antioxidants by OJE treatment may be a legitimate strategy for decreasing oxidative stresses in the kidney.

Hypoxia-ischemia leads to serious neuronal damage in some brain regions and is a strong risk factor for stroke. The aim of this study was to investigate the neuroprotective effect of tanshinone I (TsI) derived from Danshen (Radix Salvia miltiorrhiza root extract) against neuronal damage using a mouse model of cerebral hypoxia-ischemia. Brain infarction and neuronal damage were examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin histochemistry, and Fluoro-Jade B histofluorescence. Pre-treatment with TsI (10 mg/kg) was associated with a significant reduction in infarct volume 1 day after hypoxia-ischemia was induced. In addition, TsI protected against hypoxia-ischemia-induced neuronal death in the ipsilateral region. Our present findings suggest that TsI has strong potential for neuroprotection against hypoxic-ischemic damage. These results may be used in research into new anti-stroke medications.
Animals ; Brain ; Brain Infarction ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; Eosine Yellowish-(YS) ; Fluoresceins ; Hematoxylin ; Hypoxia-Ischemia, Brain ; Mice ; Neurons ; Neuroprotective Agents ; Risk Factors ; Salvia miltiorrhiza ; Stroke ; Tetrazolium Salts

Clonorchis sinensis is a carcinogenic human liver fluke that promotes hepatic inflammatory environments via direct contact or through their excretory-secretory products (ESPs), subsequently leading to cholangitis, periductal fibrosis, liver cirrhosis, and even cholangiocarcinoma (CCA). This study was conducted to examine the host inflammatory responses to C. sinensis ESPs and their putative protein components selected from C. sinensis expressed sequenced tag (EST) pool databases, including TGF-β receptor interacting protein 1(CsTRIP1), legumain (CsLeg), and growth factor binding protein 2 (CsGrb2). Treatment of CCA cells (HuCCT1) with the ESPs or bacterial recombinant C. sinensis proteins differentially promoted the secretion of proinflammatory cytokines (IL-1β, IL-6, and TNF-α) as well as anti-inflammatory cytokines (IL-10, TGF-β1, and TGF-β2) in a time-dependent manner. In particular, recombinant C. sinensis protein treatment resulted in increase (at maximum) of ~7-fold in TGF-β1, ~30-fold in TGF-β2, and ~3-fold in TNF-α compared with the increase produced by ESPs, indicating that CsTrip1, CsLeg, and CsGrb2 function as strong inducers for secretion of these cytokines in host cells. These results suggest that C. sinensis ESPs contribute to the immunopathological response in host cells, leading to clonorchiasis-associated hepatobiliary abnormalities of greater severity.
Carrier Proteins ; Cholangiocarcinoma ; Cholangitis ; Clonorchis sinensis ; Cytokines ; Fasciola hepatica ; Fibrosis ; Humans ; Interleukin-6 ; Liver Cirrhosis

BACKGROUNDOenanthe javanica (O. javanica) has been known to have high antioxidant properties via scavenging reactive oxygen species. We examined the effect of O. javanica extract (OJE) on antioxidant enzymes in the rat liver.

METHODSWe examined the effect of the OJE on copper, zinc-superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), catalase (CAT), and glutathione peroxidase (GPx) in the rat liver using immunohistochemistry and western blot analysis. Sprague-Dawley rats were randomly assigned to three groups; (1) normal diet fed group (normal-group), (2) diet containing ascorbic acid (AA)-fed group (AA-group) as a positive control, (3) diet containing OJE-fed group (OJE-group).

RESULTSIn this study, no histopathological finding in the rat liver was found in all the experimental groups. Numbers of SOD1, SOD2, CAT, and GPx immunoreactive cells and their protein levels were significantly increased in the AA-fed group compared with those in the normal-group. On the other hand, in the OJE-group, numbers of SOD1, SOD2, CAT, and GPx immunoreactive cells in the liver were significantly increased by about 190%, 478%, 685%, and 346%, respectively, compared with those in the AA-group. In addition, protein levels of SOD1, SOD2, CAT, and GPx in the OJE-group were also significantly much higher than those in the AA-group.

CONCLUSIONOJE significantly increased expressions of SOD1 and SOD2, CAT, and GPx in the liver cells of the rat, and these suggests that significant enhancements of endogenous enzymatic antioxidants by OJE might be a legitimate strategy for decreasing oxidative stresses in the liver.

Animals ; Antioxidants ; metabolism ; Ascorbic Acid ; pharmacology ; Catalase ; metabolism ; Glutathione Peroxidase ; metabolism ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; metabolism ; Male ; Oenanthe ; chemistry ; Oxidative Stress ; drug effects ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

In the present study, we investigated the effect of Tetaus toxin (TeT) on cell proliferation and neuroblast differentiation using specific markers: 5-bromo-2-deoxyuridine (BrdU) as an exogenous marker for cell proliferation, Ki-67 as an endogenous marker for cell proliferation and doublecortin (DCX) as a marker for neuroblasts in the mouse hippocampal dentate gyrus (DG) after TeT treatment. Mice were intraperitoneally administered 2.5 and 10 ng/kg TeT and sacrificed 15 days after the treatment. In both the TeT-treated groups, no neuronal death occurred in any layers of the DG using neuronal nuclei (NeuN, a neuron nuclei maker) and Fluoro-Jade B (F-J B, a high-affinity fluorescent marker for the localization of neuronal degeneration). In addition, no significant change in glial activation in both the 2.5 and 10 ng/kg TeT-treated-groups was found by GFAP (a marker for astrocytes) and Iba-1 (a marker for microglia) immunohistochemistry. However, in the 2.5 ng/kg TeT-treated-group, the mean number of BrdU, Ki-67 and DCX immunoreactive cells, respectively, were apparently decreased compared to the control group, and the mean number of each in the 10 ng/kg TeT-treated-group was much more decreased. In addition, processes of DCX-immunoreactive cells, which projected into the molecular layer, were short compared to those in the control group. In brief, our present results show that low dosage (10 ng/kg) TeT treatment apparently decreased cell proliferation and neuroblast differentiation in the mouse hippocampal DG without distinct gliosis as well as any loss of adult neurons.
Adult ; Animals ; Bromodeoxyuridine ; Cell Proliferation ; Dentate Gyrus ; Exotoxins ; Fluoresceins ; Gliosis ; Humans ; Immunohistochemistry ; Mice ; Neurogenesis ; Neurons ; Tetanus ; Tetanus Toxin