Periprosthetic Fracture after Hook Plate Fixation in Neer Type II Distal Clavicle Fracture: A Report of 3 Cases

Hook plate fixation is a treatment method for the displaced distal clavicle fracture with favorable results regarding bone union and shoulder function, however possible complications include impingement syndromes, subacrormial erosions, acromial fractures, and periprosthetic fractures. In this report, we observed 3 cases of periprosthetic fracture after hook plate fixation. All cases of periprosthetic fractures were initiated at the medial end screw holes. The causes of these periprosthetic fractures appeared to be the off centered fixation of medial end screws near the anterior or posterior cortex which were specific during operations with hook plates with more than 6 holes and the increased stress on the medial end screw by over-reduced or inferiorly reduced position of the distal end of the clavicle by the hook plate.

Sex determination using upper limb bones in Korean populations / 대한해부학회지

The purpose of this research is to establish metric standards for the determination of sex from the upper limb bones of Korean. We took a set of eleven measurements on each of 175 right sides of adult skeletons chosen at Korean sample. Classification accuracy dropped only one or two individuals when only vertical head diameter of humerus is used. Variables in relation with maximal length were less accurate than head diameter of humerus. Two variables were selected by the stepwise procedure: maximal length of humerus, vertical head diameter of humerus. The combined accuracy was 87%. This study of modern Korean skeletons underscores the need for population-specific techniques, not only for medicolegal investigations, but also for the study of population affinities and factors affecting bone configurations.

Pes anserinus and anserine bursa: anatomical study / 대한해부학회지

This study investigated the boundary of anserine bursa with the recommended injection site and shape on the insertion area of pes anserinus (PA), with the aim of improving clinical practice. Eighty six legs from 45 Korean cadavers were investigated. The mixed gelatin solution was injected to identify the shape of anserine bursa, and then the insertion site of the PA tendons was exposed completely and carefully dissected to identify the shape of the PA. The sartorius was inserted into the superficial layer and gracilis, and the semitendinosus was inserted into the deep layer on the medial surface of the tibia. The number of the semitendinosus tendons at the insertion site varied: 1 in 66% of specimens, 2 in 31%, and 3 in 3%. The gracilis and semitendinosus tendons were connected to the deep fascia of leg. Overall, the shape of the anserine bursa was irregularly circular. Most of the anserine bursa specimens reached the proximal line of the tibia, and some of the specimens reached above the proximal line of the tibia. In the medial view of the tibia, the anserine bursa was located posteriorly and superiorly from the tibia's midline, and it followed the lines of the sartorius muscle. The injection site for anserine bursa should be carried out at 20degrees from the vertical line medially and inferiorly, 15 or 20 mm deeply, and at the point of about 20 mm medial and 12 mm superior from inferomedial point of tibial tuberosity.

Pes anserinus and anserine bursa: anatomical study / 대한해부학회지

This study investigated the boundary of anserine bursa with the recommended injection site and shape on the insertion area of pes anserinus (PA), with the aim of improving clinical practice. Eighty six legs from 45 Korean cadavers were investigated. The mixed gelatin solution was injected to identify the shape of anserine bursa, and then the insertion site of the PA tendons was exposed completely and carefully dissected to identify the shape of the PA. The sartorius was inserted into the superficial layer and gracilis, and the semitendinosus was inserted into the deep layer on the medial surface of the tibia. The number of the semitendinosus tendons at the insertion site varied: 1 in 66% of specimens, 2 in 31%, and 3 in 3%. The gracilis and semitendinosus tendons were connected to the deep fascia of leg. Overall, the shape of the anserine bursa was irregularly circular. Most of the anserine bursa specimens reached the proximal line of the tibia, and some of the specimens reached above the proximal line of the tibia. In the medial view of the tibia, the anserine bursa was located posteriorly and superiorly from the tibia's midline, and it followed the lines of the sartorius muscle. The injection site for anserine bursa should be carried out at 20degrees from the vertical line medially and inferiorly, 15 or 20 mm deeply, and at the point of about 20 mm medial and 12 mm superior from inferomedial point of tibial tuberosity.

HLA-DRB1 Study of DNA from Ancient Human Skeleton by Sequence-based Typing / 체질인류학회지

The analysis of ancient human DNA is increasingly used recently in the study of anthropology and human evolution. Although mitochondrial DNA and Y chromosomal DNA has commonly been the target in the field of human DNA study, HLA analysis of ancient human DNA is extremely rare. This study aimed to develop the PCR method of ancient human DNA for analyzing the sequence of HLA. Authors established a new method for HLA-DRB1 analysis by sequence-based typing. Alleles of HLA-DRB1 were analyzed and typed by sequencing with DNA of ancient human skeletons from Korea and Mongolia 3000-500 years ago. The types of HLA-DRB1 were determined by comparing the sequences with those of HLA database (http://www. ebi.ac.uk/Tools/blast2/nucleotide.html). The alleles of HLA-DRB1 of ancient human DNA from Korea and Mongolia were classified by types. The frequencies of HLA-DRB1 types of Mongolia were also presented according to the geography such as West, Central, East, and North. In summary, our method was successful in the analyzing the type of HLA-DRB1 from DNA of ancient human bones. Authors anticipate that many researchers could do their research in a better way to get the genetic information for the kinship analysis between individuals or communities from ancient human bones.

Acute Testis Toxicity of Bisphenol A Diglycidyl Ether in Sprague-Dawley Rats / 예방의학회지

OBJECTIVES: Bisphenol A diglycidyl ether (BADGE) is a liquid compound obtained by condensation of two molecules of epichlorohydrin with one molecule of bisphenol A. General and reproductive toxicity with BADGE has been reported higher than 1000 mg/kg/day. This study was performed to show the effects of acute exposure to BADGE below 1000 mg/kg/day on the testis in adult male rats. METHODS: BADGE was administered by gastric lavage in a single dose of 500, 750, 1000, and 2000 mg/kg/day in 8-week old male SPF Sprague-Dawley rats. The right testis was processed for light microscopic analysis. The left testis was homogenized and spermatids were counted to determine the daily sperm production and daily abnormal sperm production. The sperm count, sperm motility, and incidence of abnormal sperm were estimated in the epididymis. In testicular sections, the seminiferous tubules were observed for qualitative changes. The progression of spermatogenesis was arbitrarily classified as full-matured, maturing, and immature. The specimen slide was observed at 3 points and 10 seminiferous tubules were evaluated at each point. RESULTS: The male rats exposed to single oral dose of BADGE at 750, 1000, and 2000 mg/kg/day were significantly increased the number of immature and maturing sperm on the testis. There were no significant differences with respect to sperm head count, sperm motility, and sperm abnormality in the BADGE treatment groups. CONCLUSIONS: These results suggest that single oral exposure of BADGE 750 mg/kg/day can affect adult male testis development.

A Kinship Analysis of Ancient Human Bones and Teeth from Mongolia / 체질인류학회지

The kinship was analyzed genetically on the three 2000 year old ancient human bones and teeth excavated in Mongolia. The samples were processed in a clean room to prevent the contamination from modern human DNA. The DNA extraction and purification was done with ion-exchange column kit (Qiagen G-tip 20G, USA). The PCR was done with purified DNAs from ancient human bones for paternal Y-SNP haplogroup, maternal mtDNA haplogroup, and autosomal short tandem repeats (STR). Two samples belonged to the maternal D major haplogroup, which is one of the most frequent types in the present North East Asia. One of them, showing male genotype, belonged to the paternal C major haplogroup, which is also one of the most frequent types in the present North East Asia. The remaining one belonged to the paternal R major haplogroup, frequent in the present Europe, and the maternal U haplogroup, frequent in the present Europe and East Mediterranean. The repeated results were consistent in the autosomal STR PCR. The STR data were analyzed with DNA-VIEW program (http://www.dna-view.com), which showed no close kinship among the three ancient humans. Our method was successful in the analyzing kinship among ancient human bones, which has been possible in few restricted laboratories in the World. Authors anticipate that many researchers could do their research in a better way to get the genetic information from ancient human bones.

Comparison of SNP Occurred in Coding Regions of Mitochondrial DNA Analyzed from Korean and Mongolian / 체질인류학회지

Even though mitochondrial DNA analysis is performed in the field of molecular genetics, differences of the results exist regarding which nucleotide positions are analyzed. In this study, we strategically analyzed to find ethnic specific SNP of coding regions of mitochondrial DNA of Korean and Mongolian. Mitochondrial DNA was analyzed with PCR amplification and sequencing with 112 blood samples of Korean and 92 blood samples of Mongolian. As a result, the mutation which commonly appears both in Korean and Mongolian population is 17 nucleotide positions, and the one that shown in the only Korean is 13 nucleotide positions, the one that shown in the only Mongolian 26 nucleotide positions. However, it was thought as individual variation as most mutations are shown in a sample. Among them, it appears as 9% substitution rate in 10397, 4850 nucleotide position of Korean, whereas 12.3% or 15% substitution rate in 5108, 9950 nucleotide positions of Mongolian, respectively. Beside, we observed high level of heteroplasmy in 3546, 3553 nucleotide positions. Therefore, we suggest that these regions might be novel genetic markers for dividing mitochondrial haplogroup of Korean and Mongolian population, but additional analysis needs on several nucleotide positions in huge samples as analyzing on restricted nucleotide positions using restricted DNA samples.

Heteroplasmy of Mitochondrial DNA Shown in Korean and Mongolian / 체질인류학회지

As characterization of mitochondrial DNA (mtDNA) shows maternal inheritance and exists as more than thousands copies per cell, it is widely used for population genetics and forensic scientific field. However, mitochondrial DNA study has difficulties because heteroplasmy of mtDNA is being reported from coding and control region. In this study, we have analyzed 200 samples to examine heteroplasmy in mitochondrial DNA of Korean and Mongolian. The control region and coding region in mtDNA of blood from Koreans and Mongolians were analyzed with PCR amplication and sequencing. As a result, several heteroplasmy was observed from total 10 positions including 5 positions in coding region and 5 positions in control region, respectively. Moreover, it showed more than one heteroplasmy in coding region from 6 samples in Korean and 17 samples in Mongolian. Interestingly, heteroplasmy at 5178 position was shown in 6 samples among 23 samples. Considering that the position is important for deciding haplogroup D, we suggest that additional analysis on 4883 position needs for correct haplogrouping. Beside, we also found heteroplasmy in the other positions of 204, 4853, or 16249. Therefore, we suggest that it is required of combinatory analysis on several key nucleotide positions to obtain good results when determining mitochondrial haplogroups.

Establishment of PCR Reaction Condition for Highly Successful Ancient DNA PCR / 체질인류학회지

The ancient bone DNA analysis essentially requires PCR amplification of the targeting genes of study due to the limitation of the ancient bone sample and DNA amounts. In contrast to the fresh living human DNA, it is common to face failing in amplifying the poorly preserved ancient DNA after death. Therefore, the optimized PCR methods appropriate for ancient DNA are required. However, there is no report to date that a systemic investigation of enhanced PCR amplification methods suitable for ancient samples has been conducted Approximately 500~3,300-year-old Korean and Mongolian ancient bones that are resistant to PCR were selected and an extensive number of PCR conditions were systematically investigated for the comparison of PCR success rates. For the PCR analysis, a mitochondrial DNA fragment as a multicopy DNA and a M175 Y chromosome biallelic marker DNA fragment as a single copy DNA that is the marker of the prevalent Y haplogroup (haplogroup O) in Korea were targeted. The identity of the amplified products were confirmed by DNA sequencing. Through this study, we established the optimized PCR conditions for the highly successful amplification of ancient bone DNAs. This estabilished method allowed for the successful amplification of mitochondrial DNAs from all the ancient bone samples tested and the amplification by 50% success rates in the amplification of M175 Y chromosome biallelic marker DNA but with the highest success rates. These results demonstrate that the optimized PCR condition will be useful for the promising ancient DNA analysis in the fields of molecular genetic anthropological studies.

Protective Effect of HSP90 on Neuronal Cell Death-induced by beta-Amyloid Peptide / 체질인류학회지

In the present study, we determined the protective mechanism of HSP90 against neuronal cell death induced by Abeta. For the evaluation of protective role of HSP90, we used human neuroblastoma SK-N-SH cell lines, examined AlamarBlue assay, Western blot analysis and immunofluorescence assay. Incubation of SK-N-SH cells with Abeta significantly induced neuronal cell death. However, HSP90 induced by mild heat shock could attenuate neuronal apoptosis in Abeta treated condition. To identify the role of HSP90, we determined localization of HSP90 in SK-N-SH cells. Interestingly, HSP90 was increased and localized in mitochondria as treatment of mild heat shock. Also, treatment or increase of HSP90 largely elevated level of Bcl-2 expression, whereas inhibition of HSP90 with HSP90 antisense oligonucleotide significantly decreased Bcl-2 expression. In contrast to Bcl-2, Bax expression was regulated independently by HSP90. Moreover, increase of HSP90 could attenuate collapse of mitochondrial membrane potential induced by Abeta. However, HSP90 antisense oligonucleotide largely increase breakdown of mitochondrial membrane potential induced by Abeta. These data suggest that HSP90 as chaperone protein significantly attenuates neuronal damage and protects neuroanl cells from neurotoxin such as Abeta.

Immunohistochemical Study on the Distribution of Insulin-like Growth Factor Binding Protein 7 (IGFBP7) in the Central Nervous System of Adult Rats / 체질인류학회지

In the present study, we performed immunohistochemical studies to investigate the detailed distribution of insulin-like growth factor binding protein 7 (IGFBP7) in the central nervous system of adult rats. Twelve adult (4~6 month old) Sprague-Dawley rats were examined in this study. Immunohistochemistry using specific antibodies against IGFBP7 was performed in accordance with the free-floating method. In the present study, IGFBP7 immunoreactivity was observed in the cerebral cortex, hippocampus, brainstem, cerebellum and spinal cord. In the cerebral cortex, heavily stained neurons were seen in layers II-VI. In the hippocampus, pyramidal cells in CA1-3 region were strongly immunoreactive for IGFBP7. Strong immunoreactive neurons were also found in the supraoptic nucleus, paraventricular nucleus, periaqueductal gray and oculomotor nucleus. In the cerebellum, IGFBP7 immunoreactivity was prominent in the Purkinje cells and cerebellar output neurons. IGFBP7-immunoreactive neurons were prominent in the superior vestibular nucleus, cochlear nucleus, trigeminal motor nucleus, nucleus of the trapezoid, and facial nucleus. IGFBP7-immunoreactive neurons were also observed mainly in the anterior horn of the spinal cord. The first demonstration of IGFBP7 localization in the whole brain may provide useful data for the future investigations on the structural and functional properties of IGFBP7.

Differential Expression of Neuronal Death-related Factors in Aged Rat Cerebellum / 체질인류학회지

In the present study, we investigated the expression of apoptosis-associated proteins in the cerebellum of aged rats: IGF-I receptor (IGF-IR), nitrotyrosine (NT), p53, key pro-apoptotic gene ICH-1 (caspase-2), c-Fos and Bcl-2 family members (Bcl-2 and Bax). Twelve adult (4~6 month old) and 15 aged (24~29 month old) Sprague-Dawley rats were examined in this study. We performed immunohistochemical staining, in situ hybridization and densitometric measurement using a NIH image program (Scion Image) to determine the staining density. In adult rats, there were no immunoreactivities for insulin-like growth factor-I receptor (IGF-IR), nitrotyrosine (NT) or p53 in any region of cerebellum. However, IGF-IR immunoreactivity was found in some Purkinje cells in aged rat cerebellum. The prominent staining of NT or p53 was also localized in the Purkinje cell layer in aged rats. A high density of ICH-1 (caspase-2) immunoreactivity was observed in the molecular and Purkinje cell layers in aged rats. Immunoreactivity for c-Fos was significantly decreased in the granule cells in aged rats. Positive signal for bcl-2 was significantly decreased in the Purkinje cells and granule cells of aged rats. The most intense staining for Bax was observed in the soma of Purkinje cells of adult rats. However, Bax immunoreactivity was not changed in any layers in the cerebellar cortex of aged rats. In conclusion, this study provides the first morphological data concerning the differential regulation of apoptosisrelated genes in rat cerebellum during aging.

Immunohistochemical Study on the Distribution of Kv1.2 in Aged Rat Brain / 체질인류학회지

In the present study, we demonstrated age-related changes in Kv1.2 immunoreactivity in the rat brain for the first time. Twelve adult (4~6 month old) and 15 aged (20~29 month old) Sprague-Dawley rats were examined in this study. Immunohistochemistry was performed in accordance with the free-floating method, and densitometric measurement using a NIH image program (Scion Image) determined the staining density. In the cerebral cortex of aged rats, there was a significant increase in the number of Kv1.2-immunoreactive neurons in the cingulate cortex, infralimbic cortex and piriform cortex, compare to adult rats. In the hippocampal CA1-3 regions, moderate Kv1.2 immunoreactivity was found in the cell bodies and processes of some medium to large-sized neurons in aged rats. The intensity was increased in the cell bodies of Kv1.2-positive neurons in the amygdala of aged rats, whereas the number of immunoreactive neurons was not significantly increased. It was noteworthy that age-related changes in Kv1.2-immunoreactive neurons were prominent in the facial nuclei, raphe magnus nuclei, and pontine and medullary reticular formation. Although the present study has not addressed multiple mechanisms contributing to neuronal degeneration during aging, the first demonstration of age-related changes in Kv1.2 immnuoreactivity may offer a comprehensive understanding of the pathophysiology of aging and age-related neurodegenerative diseases such as Alzheimer's disease.

Changes of Tight Junction and Epithelial Cells after Treatment of Cadmium / 체질인류학회지

Cadmium (Cd) affects cell proliferation, differentiation, apoptosis and other cellular activities and can cause numerous molecular lesions that would be relevant to carcinogenesis. The mechanism of adverse effects of Cd has been poorly understood and, especially on the tight junction. Since there is rare information about the effect of Cd on tight junction protein, we here investigated whether Cd can alter the localization of the proteins. This study examined Cd effects on of tight junction (occludin, ZO-1, and ZO-2) using MDCK cell culture. The change of MDCK cell and tight junction was investigated after treatment of cadmium with phase contrast microscopy, TEER, cell viability, Transmission electron microscopy and confocal laser microscopy. After treatment of cadmium, transendothelial electrical resistance decreased with time and concentration dependent manner. AlamarBlue assay revealed that decreased cell viability also decreased with time and concentration dependent manner. The tight junction moved down between intercellular spaces with decreased density and the cellular thickness around cell junctions decreased with increasing concentration and exposure time of CdCl2. The MDCK cells eventually showed cell death with. Confocal laser microscopy revealed that immunofluorescent reaction of occludin, ZO-1 and ZO-2 decreased. Occludin, ZO-1 and ZO-2 were disrupted at tight junction. These data suggest that after treatment of Cd, increased permeability of MDCK cell monolayer increased. This might be accompanied with disruption of occludin, ZO-1 and ZO-2.

Immunohistochemical Study on the Distribution of Glycogen Synthase Kinase (GSK) 3beta in the Central Nervous System of SOD1G93A Transgenic Mice / 체질인류학회지

In the present study, we investigated influences of glycogen synthase kinase (GSK) 3beta on the development and/or progression of amyotrophic lateral sclerosis (ALS). We used transgenic mice expressing a human Cu/Zn superoxide dismutase mutant (SOD1G93A) as an in vivo model of ALS and examined expressional changes of GSK3beta immunohistochemically in the spinal cord, brain stem and cerebellum. With these experiments we demonstrate that the neurons in these regions of symptomatic SOD1G93A transgenic mice showed increased GSK3beta immunoreactivities compared with wild-type SOD1 transgenic mice. In contrast to symptomatic SOD1G93A transgenic mice, few GSK3beta immunoreactivity changes were detected in 8w- and 13w-old presymptomatic SOD1G93A transgenic mice. These data suggest the possibility that GSK3 functions as a modulating factor of apoptosis-related alterations in ALS and that GSK3beta exert differential functions in the development and/or progression of ALS. But the exact functional significances of these changes require further elucidation.

Development of Ancient DNA Isolation Method for Improved PCR Amplification / 체질인류학회지

Ancient DNA analyses are widely used for evolutionary and phylogenetic study of mankind in anthropology and archeology. However, the DNA extraction from particularly poorly preserved ancient human samples is often unsuccessful in these analyses. In the present study, to improve the success rate of ancient DNA analysis, we introduced a high grade ancient DNA purification method using ion-exchange columns. We compared the success rate of ancient DNA analysis of this new method with that of the two methods that have been used for ancient DNA extraction, GENECLEAN(R) kit (Qbiogene) and Qiaquick column (Qiagen). Twelve ancient bone samples from Korea and Mongolia that are about 500 to 5,000 years old by an archeological estimation were used. As the DNA analysis methods, polymerase chain reaction (PCR) methods for the amplification of a mitochondrial DNA HV1 segment, a male sex determination marker DNA and M175 marker DNA that is used for the determination of O haplogroup of Y chromosome that is reportedly a common one in modern Korean people. The method developed in this study remarkably increased the success rate of DNA analysis compared with the other two methods. Using the GENECLEAN(R) kit, only two samples were amplifiable for the mitochondrial DNA, no samples for the male sex determination marker and M175 marker DNAs. Using the Qiaquick columns, nine samples were amplifiable for mitochondirial DNA, nine samples for male sex determination marker and six samples for M175 marker. The developed method allowed for the amplification of mitochondrial DNA from all samples, male sex determination marker from eight samples and M175 marker from eight samples. The results demonstrate that ion-exchange columns can be useful for the improved ancient DNA extraction in anthropology and archeology.

Comparison between Morphological Sex and Genotype Sex of Uzbekistan Ancient Bones Using Improved Amelogenin PCR Amplication Method / 체질인류학회지

Determination of male and female is important in anthropology, archeology and forensic science. This study was designed to compare genotype sex of improved amelogenin PCR amplication method with morphological sex of ancient human bones. Sixty human skulls which lived from the Bronze Age to twenties centuries and excavated in Uzbekistan were used in this study. Morphological sex was determined by Uzbekistan scientist, and genotype sex was determined by improved amelogenin PCR amplication developed in this study. Among 20 morphological males, 13 samples (65%) were genotypical male. Among 40 morphological females, 20 samples (50%) were genotypical male. In conclusion, morphological method might be inadequate for sex determination of ancient bones. The improved amelogenin PCR method will be useful in sex determination of ancient bones.

Study on the Mitochondrial Dysfunction by p53 Regulation in Ceramide-induced Neuronal Cell Death / 체질인류학회지

Ceramide induces cell death in a dose- and time-dependent manner in neuroblastoma SK-N-SH cells. To investigate the mechanism of SK-N-SH cell death by C2-ceramide, morphological features and Hoechst 33258 staining were analyzed. In these morphlogic study the cell death by ceramide showed typical apoptotic features, nuclear condensation, fragmentation, and membrane blebbing. Ceramide-induced apoptosis was accompanied by nuclear accumulation of p53. Inhibition of p53 expression with p53 antisense oligonucleotides inhibited apoptosis evoked by ceramide. Also, ceramide induced mitochondrial event, collapse of mitochondrial membrane potential (delta psi m) and interestingly, inhibition of p53 attenuated collapse of mitochondrial membrane potential, suggests that ceramide induces mitochondrial dysfunction through upregulation of p53 expression. These results suggest that ceramide-induced apoptosis is dependent upon increase in cellular p53 levels which play a critical role in the regulation of apoptotic cell death and p53 modulates mitochondrial function such as mitochondrial membrane potential level.

Alpha-tocopherol Prevents H2O2-induced Tight Junction Occludin Disruption in Blood-Brain Barrier / 체질인류학회지

Vitamin E is the most important lipid-soluble antioxidant in humans. Although alpha-tocopherol is suggested that it has protective effect from many diseases, little is known about the prevention of occludin alteration in tight junction of blood-brain barrier (BBB) under pathologic insults producing reactive oxygen species (ROSs). In this study, the effects of alpha-tocopherol on H2O2-induced tight junction occludin were studied. Primary culture of rat brain microvessel endothelial cells was investigated with confocal microscopy, Western blot, and cell viability assay. Alpha-tocopherol had no apparent cytotoxicity up to 2.8 mM. The preincubation with alpha-tocopherol suppressed the H2O2-induced cytotoxicity in Alamar Blue assay and phase contrast microscopy. In confocal laser microscopy and Western blot, H2O2-induced loss of occludin was suppressed by preincubation with alpha-tocopherol. The present findings provide evidence that alpha-tocopherol may be beneficial for cellular protection from pathologic insults. Since alpha-tocopherol was demonstrated to have far fewer adverse effects, it would become a noteworthy nutrient or drug for the treatment of neurodegenerative diseases.