OBJECTIVETo explore action mechanism of acupuncture and moxibustion for Alzheimer's disease (AD) to provide evidence for prevention and treatment with acupuncture and moxibustion on AD in clinic.

METHODSEighty SPF-grade male Wistar rats, (200 +/- 20) g, were randomly divided into a normal group, a sham-operation group, a model group and a treatment group, 20 cases in each one. The model was duplicated with injection of Abeta1-42 in rats' hippocampus. Expect the treatment group, the rest groups were treated with regular feeding after respective intervention. The treatment group was treated with acupuncture and moxibustion at "Baihui" (GV 20) and "Shenshu" (BL 23), once a day, seven days as a treatment course and totally for two courses. There was one day of interval between the courses. The immunohistochemistry and quantitative RT-PCR methods were applied to test level of Abeta-binding alcohol dehydrogense (ABAD) and cytochrome oxidase IV (COX IV) in hippocampal neurons mitochondria.

RESULTSAcupuncture and moxibustion could reduce effectively level of ABAD and improve activity of COX IV in hippocampal neurons mitochondria in the treatment group, which has statistical significance compared with that in the model group (P < 0.01) and no statistical significance compared with that in the normal group and sham-operation group (P > 0.05). This indicated that acupuncture and moxibustion could effectively suppress overexpression of ABAD, improve activity of COX IV and reduce leak of reactive oxygen species, which could improve metabolic disturbance of mitochondria energy to achieve the goal of prevention and treatment of AD.

CONCLUSIONThe prevention and treatment of AD with acupuncture and moxibustion could be related with suppressing overexpression of ABAD and improving activity of COX IV in hippocampal neurons mitochondria to improve mitochondria energy metabolism.

3-Hydroxyacyl CoA Dehydrogenases ; genetics ; metabolism ; Acupuncture Therapy ; Alzheimer Disease ; enzymology ; metabolism ; therapy ; Animals ; Electron Transport Complex IV ; genetics ; metabolism ; Energy Metabolism ; Hippocampus ; cytology ; enzymology ; metabolism ; Humans ; Male ; Mitochondria ; enzymology ; metabolism ; Moxibustion ; Neurons ; enzymology ; metabolism ; Rats ; Rats, Wistar

There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.
Animals ; Base Sequence ; DNA, Mitochondrial/genetics ; Electron Transport Complex IV/*genetics ; Genetic Markers/genetics ; Leeches/*classification/*genetics ; Mitochondria/enzymology/genetics ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 18S/*genetics ; Republic of Korea ; Sequence Alignment ; Sequence Analysis, DNA

Diphyllobothrium latum and Diphyllobothrium nihonkaiense are the 2 reported main causes of human diphyllobothriasis in the Republic of Korea. However, the differentiation of these 2 species based on morphologic features alone is difficult. The authors used nucleotide sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene to diagnose Diphyllobothrium spp. Two patients visited the emergency room at Kyungpook National University Hospital on 3 April and 12 April 2013, respectively, with fragments of parasites found while defecating. The parasites were identified as Diphyllobothrium spp. based on morphologic characteristics, and subsequent cox1 gene sequencing showed 99.9% similarity (1,478/1,480 bp) with D. nihonkaiense. Our findings support the hypothesis that D. nihonkaiense is a dominant species in Korea.
Adult ; Animals ; Anthelmintics/therapeutic use ; Base Sequence ; DNA, Helminth/*genetics ; Diphyllobothriasis/*diagnosis/drug therapy ; Diphyllobothrium/*genetics ; Electron Transport Complex IV/*genetics ; Humans ; Male ; Middle Aged ; Mitochondria/enzymology ; Phylogeny ; Praziquantel/therapeutic use ; Republic of Korea ; Sequence Analysis, DNA ; Young Adult

Taeniasis has been known as one of the prevalent parasitic infections in Korea. Until recently, Taenia saginata had long been considered a dominant, and widely distributed species but epidemiological profiles of human Taenia species in Korea still remain unclear. In order to better understand distribution patterns of human Taenia tapeworms in Korea, partial nucleotide sequences of mitochondrial cox1 and ITS2 (internal transcribed spacer 2) were determined, along with morphological examinations, on 68 Taenia specimens obtained from university museum collections deposited since 1935. Genomic DNA was extracted from formalin-preserved specimens. Phylogenetic relationships among the genotypes (cox1 haplotype) detected in this study were inferred using the neighbor-joining method as a tree building method. Morphological and genetic analyses identified 3 specimens as T. solium, 51 specimens as T. asiatica, and 14 specimens as T. saginata. Our results indicate that all 3 Taenia tapeworms are sympatrically distributed in Korea with T. asiatica dominating over T. saginata and T. solium.
Animals ; Base Sequence ; Cestode Infections/epidemiology/parasitology ; Cyclooxygenase 1/genetics/metabolism ; DNA, Ribosomal Spacer/genetics ; Gene Expression Regulation ; Humans ; Korea/epidemiology ; Mitochondria/enzymology ; Molecular Sequence Data ; Phylogeny ; Taenia/*classification/*genetics ; Time Factors

We designed the specific primers and TaqMan probes targeting cytochrome b genes of mitochondrial DNA from bovine, goat and sheep. We used different fluorescents to label the probes. After optimization of reaction conditions, we set up a multiplex fluorescent real-time PCR method to detect bovine, goat and sheep derived materials, simultaneously. We finished the detection tests of 17 kinds of animal DNA and 200 DNA samples from different sources with the developed method and the National Standard GB/T 20190Y-2006 routine PCR method. The coincidence rate of these two methods was 100%. Without electrophoresis or restriction digestion, the developed method could reduce the test time to one third as routine PCR and identify three kinds of animal derived materials including bovine, goat and sheep in one reaction. The developed method was approximately 10 times more sensitive than routine PCR, and was applicable to identifications of bovine, goat and sheep derived materials in feed stuff, meat, milk, pelt and grease, etc. The study showed that the developed real-time PCR method is a rapid, sensitive and efficacious detection assay for bovine, goat and sheep derived materials in animal products.
Animal Feed ; analysis ; Animals ; Cattle ; Cytochromes b ; genetics ; DNA Primers ; DNA, Mitochondrial ; analysis ; genetics ; Food Contamination ; analysis ; Genes, Mitochondrial ; Goats ; Meat ; analysis ; Mitochondria, Muscle ; enzymology ; genetics ; Polymerase Chain Reaction ; methods ; Sheep

The parasite Plasmodium falciparum causes severe malaria and is the most dangerous to humans. However, it exhibits resistance to their drugs. Farnesyltransferase has been identified in pathogenic protozoa of the genera Plasmodium and the target of farnesyltransferase includes Ras family. Therefore, the inhibition of farnesyltransferase has been suggested as a new strategy for the treatment of malaria. However, the exact functional mechanism of this agent is still unknown. In addition, the effect of farnesyltransferase inhibitor (FTIs) on mitochondrial level of malaria parasites is not fully understood. In this study, therefore, the effect of a FTI R115777 on the function of mitochondria of P. falciparum was investigated experimentally. As a result, FTI R115777 was found to suppress the infection rate of malaria parasites under in vitro condition. It also reduces the copy number of mtDNA-encoded cytochrome c oxidase III. In addition, the mitochondrial membrane potential (DeltaPsim) and the green fluorescence intensity of MitoTracker were decreased by FTI R115777. Chloroquine and atovaquone were measured by the mtDNA copy number as mitochondrial non-specific or specific inhibitor, respectively. Chloroquine did not affect the copy number of mtDNA-encoded cytochrome c oxidase III, while atovaquone induced to change the mtDNA copy number. These results suggest that FTI R115777 has strong influence on the mitochondrial function of P. falciparum. It may have therapeutic potential for malaria by targeting the mitochondria of parasites.
Antimalarials/*pharmacology ; Enzyme Inhibitors/*pharmacology ; Farnesyltranstransferase/*antagonists & inhibitors/genetics/*metabolism ; Humans ; Malaria, Falciparum/drug therapy/*parasitology ; Mitochondria/*drug effects/metabolism ; Plasmodium falciparum/drug effects/*enzymology/genetics ; Protozoan Proteins/*antagonists & inhibitors/genetics/metabolism ; Quinolones/*pharmacology

We analyzed the sequence of vertebrate molecular marker genes, then we selected the mitochondrial DNA (mtDNA) 16S rRNA gene as marker gene. In order to detect four kinds of animal-derived ingredients, which including bovine, goat, pig and chicken. We utilized a pair of universal primers, designed four sets of species-specific microarray probes and two pairs of quality control probes. We optimized the PCR amplifications and hybridization conditions, therefore these four kinds of animal-derived ingredients could be rapid and accurate detected by this approach. The detection limits were all reaches 1 pg. We established the detection platform of these four kinds of animal-derived ingredients. This universal PCR-microarray assay provides a new method for the identification of animal-derived ingredients in the import-export field.
Animal Feed ; analysis ; Animals ; Cattle ; Chickens ; DNA, Mitochondrial ; genetics ; Food Contamination ; analysis ; Goats ; Meat ; analysis ; Microarray Analysis ; methods ; Mitochondria, Muscle ; enzymology ; genetics ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Sensitivity and Specificity ; Swine

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

A subset of patients of amyotrophic lateral sclerosis (ALS) present with mutation of Cu/Zn superoxide dismutase 1 (SOD1), and such mutants caused an ALS-like disorder when expressed in rodents. These findings implicated SOD1 in ALS pathogenesis and made the transgenic animals a widely used ALS model. However, previous studies of these animals have focused largely on motor neuron damage. We report herein that the spinal cords of mice expressing a human SOD1 mutant (hSOD1-G93A), besides showing typical destruction of motor neurons and axons, exhibit significant damage in the sensory system, including Wallerian-like degeneration in axons of dorsal root and dorsal funiculus, and mitochondrial damage in dorsal root ganglia neurons. Thus, hSOD1-G93A mutation causes both motor and sensory neuropathies, and as such the disease developed in the transgenic mice very closely resembles human ALS.
Amyotrophic Lateral Sclerosis/enzymology/*pathology ; Animals ; Axons/*pathology ; Disease Models, Animal ; Ganglia, Spinal/pathology ; Humans ; Mice ; Mice, Transgenic ; Mitochondria/pathology ; Motor Neurons/metabolism/pathology ; Mutation ; Nerve Degeneration/*pathology ; Sensory Receptor Cells/*pathology ; Spinal Cord/*pathology ; Superoxide Dismutase/genetics/*physiology