1.Construction of eukaryotic expression plasmid pSecTag2B-msBlyS expressing mouse soluble B lymphocyte stimulator.
Chun-hua FU ; Ling TIAN ; Yu-quan WEI ; Yan-jun WEN ; Jong LI
West China Journal of Stomatology 2004;22(2):145-148
OBJECTIVEThe purpose of this study was to clone the soluble form of the mouse BlyS (msBlyS) and insert it into a eukaryotic expression vector pSecTag2B in order to further elucidat the antitumor activity induced by msBlyS expressed by the recombined plasmid pSecTag2B-msBlyS.
METHODSFull length cDNA of mouse soluble BlyS (msBlyS) was amplified by reverse transcription-PCR from total RNA of mouse spleen. The PCR product was ligated directly with linearized vector pCR2.1 supplied in the TA cloning kit. The recombined plasmid pCR2.1-msBlyS which was selected and identified using blue-white screening method and restriction map analysis and the purified original plasmid pSecTag2B were both cut by HindIII and EcoR I. The digested fragments were extracted and purified from low-melting temperature agarose and ligated by T4DNA ligase. The recombined plasmid pSecTag2B-msBlyS were isolated and identified by restricted endonuclease cutting and Sanger dideoxy DNA sequencing.
RESULTSThe sequencing data indicated that inserted msBlyS gene had correct DNA sequence and orientation.
CONCLUSIONEukaryotic expression vector pSecTag2B. Expressing mouse BlyS have successfully been cloned. This will provide us an opportunity to do further research work on BlyS.
Animals ; B-Cell Activating Factor ; Cloning, Molecular ; Epitopes, B-Lymphocyte ; genetics ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Membrane Proteins ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Polymerase Chain Reaction ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; Recombination, Genetic ; Sequence Analysis, DNA ; Spleen ; cytology ; immunology ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics
2.Endovascular stent-graft repair of aortocaval fistula complicated by the rupture of abdominal aortic aneurysm.
Su Jin SIN ; Jin Woo KIM ; Hyun Su KIM ; Yeon Ik CHOO ; Jong Jun YAN ; Hyo Seop LEE ; Jae Cheol HWANG
Korean Journal of Medicine 2004;67(Suppl 3):S746-S751
The incidence of infrarenal aneurym is about 6 percent after the age 60 years. A primary aortocaval fistula is present in less than 1% of all abdominal aortic aneurysms. Atherosclerotic abdominal aortic aneurysm account for about 90% of spontanous aortocaval fistula. The most common site of fistulation is the inferior vena cava. Until recently, surgical repair was the only method of treatment and was associated high incidence of morbidity and motality. With rapid development of aortic stent-graft technique, endovascular stent-graft repair may offer an alterative to the management of this often fatal condition. We report a case of 72-years old male with aortocaval fistula in the abdominal aorta, which was treated with endovascular stent-graft implantation. About 30 days before procedure, the patient diagnosed inferior acute myocardial infarction with triple vessel disease and also suffered from chronic obstructive pulmonary disease. After the stent-graft inserting, no further communication from aorta to inferior vena cava and improving symptoms and sign of congestive heart failure. He was discharged without complication, about 2 months after admission.
Aged
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Aneurysm
;
Aorta
;
Aorta, Abdominal
;
Aortic Aneurysm
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Aortic Aneurysm, Abdominal*
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Endovascular Procedures
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Fistula*
;
Heart Failure
;
Humans
;
Incidence
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Male
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Myocardial Infarction
;
Pulmonary Disease, Chronic Obstructive
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Rupture*
;
Vena Cava, Inferior
3.Stem Cell Dynamics in an Experimental Model of Stroke
Min Cheol LEE ; Chun Yan JIN ; Hyung Seok KIM ; Jae Hyu KIM ; Myeong Kyu KIM ; Hyoung Ihl KIM ; Young Jin LEE ; Young Jun SON ; Young Ok KIM ; Young Jong WOO
Chonnam Medical Journal 2011;47(2):90-98
We investigated the migration of endogenous neural stem cells (NSCs) toward an infarct lesion in a photo-thrombotic stroke model. The lesions produced by using rose bengal dye (20 mg/kg) with cold light in the motor cortex of Sprague-Dawley rats were also evaluated with sequential magnetic resonance imaging (MRI) from 30 minutes through 8 weeks. Migration of NSCs was identified by immunohistochemistry for nestin monoclonal antibody in the lesion cortex, subventricular zone (SVZ), and corpus callosum (CC). The contrast to noncontrast ratio (CNR) on MRI was greatest at 12 hours in DWI and decreased over time. By contrast, T1-weighted and T2-weighted images showed a constant CNR from the beginning through 8 weeks. MRI of the lesional cortex correlated with histopathologic findings, which could be divided into three stages: acute (edema and necrosis) within 24 hours, subacute (acute and chronic inflammatory cell infiltration) at 2 to 7 days, and chronic (gliofibrosis) at 2 to 4 weeks. The volume of the infarct was significantly reduced by reparative gliofibrosis. The number of nestin+ NSCs in the contralateral SVZ was similar to that of the ipsilateral SVZ in each group. However, the number of nestin+ NSCs in the ipsilateral cortex and CC increased at 12 hours to 3 days compared with the contralateral side (p<0.01) and was reduced significantly by 7 days (p<0.01). Active emigration of internal NSCs from the SVZ toward the infarct lesion may also contribute to decreased volume of the infarct lesion, but the self-repair mechanism by endogenous NSCs is insufficient to treat stroke causing extensive neuronal death. Further studies should be focused on amplification technologies of NSCs to enhance the collection of endogenous or transplanted NSCs for the treatment of stroke.
Cold Temperature
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Corpus Callosum
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Emigration and Immigration
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Immunohistochemistry
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Intermediate Filament Proteins
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Light
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Magnetic Resonance Imaging
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Models, Theoretical
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Motor Cortex
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Nerve Tissue Proteins
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Neural Stem Cells
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Neurons
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Rats, Sprague-Dawley
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Rose Bengal
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Stem Cells
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Stroke
;
Transplants
4.Systemic administration of low dosage of tetanus toxin decreases cell proliferation and neuroblast differentiation in the mouse hippocampal dentate gyrus.
Bing Chun YAN ; In Hye KIM ; Joon Ha PARK ; Ji Hyeon AHN ; Jeong Hwi CHO ; Bai Hui CHEN ; Jae Chul LEE ; Jung Hoon CHOI ; Ki Yeon YOO ; Choong Hyun LEE ; Jun Hwi CHO ; Jong Dai KIM ; Moo Ho WON
Laboratory Animal Research 2013;29(3):148-155
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
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Animals
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Bromodeoxyuridine
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Cell Proliferation
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Dentate Gyrus
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Exotoxins
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Fluoresceins
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Gliosis
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Humans
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Immunohistochemistry
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Mice
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Neurogenesis
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Neurons
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Tetanus
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Tetanus Toxin
5.Hemorrhagic fever caused by a novel tick-borne Bunyavirus in Huaiyangshan,China
Zhang YONG-ZHEN ; Zhou DUN-JIN ; Xiong YANWEN ; Chen XIAO-PING ; He YONG-WEN ; Sun QIANGZHENG ; Yu BIN ; Li JUAN ; Dai YONG-AN ; Tian JUN-HUA ; Qin XIN-CHENG ; Jin DONG ; Cui ZHIGANG ; Luo XUE-LIAN ; Li WEI ; Lu SHAN ; Wang WEN ; Peng JIN-SONG ; Guo WEN-PING ; Li MING-HUI ; Li ZHEN-JUN ; Zhang SHAOMIN ; Chen CHEN ; Wang YAN ; Jong D.de MENNO ; Xu JIANGUO
Chinese Journal of Epidemiology 2011;32(3):209-220
Background:From April to July in 2009 and 2010,unexplained severe hemorrhagic fever-like illnesses occurred in farmers from the Huaiyangshan mountains range.Methods:Clinical specimens (blood,urine,feces,and throat swabs) from suspected patients were obtained and stored.Mosquitoes and ticks in affected regions were collected.Virus was isolated from 2 patients and characterized by whole genome sequencing.Virus detection in additional patients and arthropods was done by virus-specific reverse transcription (RT) PCR.Clinical and epidemiological data of RT-PCR confirmed patients were analyzed.Results:An unknown virus was isolated from blood of two patients and from Haemaphysalis ticks collected from dogs.Whole genome sequence analysis identified the virus as a novel member of the family Bunyaviridae,most closely related to the viruses of the genus Phlebovirus within which it forms a separate lineage.Subsequently,infection was confirmed by RT-PCR in 33 of 58 suspected patients.The illness in these patients was characterized by fever,severe malaise,nausea,vomiting,and diarrhea.Prominent laboratory findings included low white cell- and platelet counts,coagulation disturbances,and elevation of liver enzymes.Hemorrhagic complications were observed in 3 cases,5 (15%) patients died.Conclusions:A novel tick-borne Bunyavirus causing life-threatening hemorrhagic fever in humans has emerged in the Huaiyangshan mountain areas of China.Further studies are needed to determine the epidemiology,geographic distribution and vertebrate animal ecology of this virus.
6.Erratum: Increased Expression of Herpes Virus-Encoded hsv1-miR-H18 and hsv2-miR-H9-5p in Cancer-Containing Prostate Tissue Compared to That in Benign Prostate Hyperplasia Tissue.
Seok Joong YUN ; Pildu JEONG ; Ho Won KANG ; Helen Ki SHINN ; Ye Hwan KIM ; Chunri YAN ; Young Ki CHOI ; Dongho KIM ; Dong Hee RYU ; Yun Sok HA ; Tae Hwan KIM ; Tae Gyun KWON ; Jung Min KIM ; Sang Heon SUH ; Seon Kyu KIM ; Seon Young KIM ; Sang Tae KIM ; Won Tae KIM ; Ok Jun LEE ; Sung Kwon MOON ; Nam Hyung KIM ; Isaac Yi KIM ; Jayoung KIM ; Hee Jae CHA ; Yung Hyun CHOI ; Eun Jong CHA ; Wun Jae KIM
International Neurourology Journal 2016;20(4):374-374
In this article, a part of fund and grant supports was omitted unintentionally.
7.Increased Expression of Herpes Virus-Encoded hsv1-miR-H18 and hsv2-miR-H9-5p in Cancer-Containing Prostate Tissue Compared to That in Benign Prostate Hyperplasia Tissue.
Seok Joong YUN ; Pildu JEONG ; Ho Won KANG ; Helen Ki SHINN ; Ye Hwan KIM ; Chunri YAN ; Young Ki CHOI ; Dongho KIM ; Dong Hee RYU ; Yun Sok HA ; Tae Hwan KIM ; Tae Gyun KWON ; Jung Min KIM ; Sang Heon SUH ; Seon Kyu KIM ; Seon Young KIM ; Sang Tae KIM ; Won Tae KIM ; Ok Jun LEE ; Sung Kwon MOON ; Nam Hyung KIM ; Isaac Yi KIM ; Jayoung KIM ; Hee Jae CHA ; Yung Hyun CHOI ; Eun Jong CHA ; Wun Jae KIM
International Neurourology Journal 2016;20(2):122-130
PURPOSE: Previously, we reported the presence of virus-encoded microRNAs (miRNAs) in the urine of prostate cancer (CaP) patients. In this study, we investigated the expression of two herpes virus-encoded miRNAs in prostate tissue. METHODS: A total of 175 tissue samples from noncancerous benign prostatic hyperplasia (BPH), 248 tissue samples from patients with CaP and BPH, and 50 samples from noncancerous surrounding tissues from these same patients were analyzed for the expression of two herpes virus-encoded miRNAs by real-time polymerase chain reaction (PCR) and immunocytochemistry using nanoparticles as molecular beacons. RESULTS: Real-time reverse transcription-PCR results revealed significantly higher expression of hsv1-miR-H18 and hsv2-miRH9- 5p in surrounding noncancerous and CaP tissues than that in BPH tissue (each comparison, P<0.001). Of note, these miRNA were expressed equivalently in the CaP tissues and surrounding noncancerous tissues. Moreover, immunocytochemistry clearly demonstrated a significant enrichment of both hsv1-miR-H18 and hsv2-miR-H9 beacon-labeled cells in CaP and surrounding noncancerous tissue compared to that in BPH tissue (each comparison, P<0.05 for hsv1-miR-H18 and hsv2- miR-H9). CONCLUSIONS: These results suggest that increased expression of hsv1-miR-H18 and hsv2-miR-H95p might be associated with tumorigenesis in the prostate. Further studies will be required to elucidate the role of these miRNAs with respect to CaP and herpes viral infections.
Carcinogenesis
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Herpesviridae
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Humans
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Hyperplasia*
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Immunohistochemistry
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MicroRNAs
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Nanoparticles
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Prostate*
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Prostatic Hyperplasia
;
Prostatic Neoplasms
;
Real-Time Polymerase Chain Reaction
8.PretreatedExtract Prevents Neuronal Death Following Transient Global Cerebral Ischemia through Increases of Superoxide Dismutase 1 and Brain-derived Neurotrophic Factor Expressions in the Gerbil Hippocampal Cornu Ammonis 1 Area.
Joon Ha PARK ; Tae-Kyeong LEE ; Bing-Chun YAN ; Bich-Na SHIN ; Ji Hyeon AHN ; In Hye KIM ; Jeong Hwi CHO ; Jae-Chul LEE ; In Koo HWANG ; Jong Dai KIM ; Seongkweon HONG ; Young Joo LEE ; Moo-Ho WON ; Il Jun KANG
Chinese Medical Journal 2017;130(15):1796-1803
BACKGROUNDGlehnia littoralis, as a traditional herbal medicine to heal various health ailments in East Asia, displays various therapeutic properties including antioxidant effects. However, neuroprotective effects of G. littoralis against cerebral ischemic insults have not yet been addressed. Therefore, in this study, we first examined its neuroprotective effects in the hippocampus using a gerbil model of transient global cerebral ischemia (TGCI).
METHODSGerbils were subjected to TGCI for 5 min. G. littoralis extract (GLE; 100 and 200 mg/kg) was administrated orally once daily for 7 days before ischemic surgery. Neuroprotection was examined by neuronal nuclear antigen immunohistochemistry and Fluoro-Jade B histofluorescence staining. Gliosis was observed by immunohistochemistry for glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1. For neuroprotective mechanisms, immunohistochemistry for superoxide dismutase (SOD) 1 and brain-derived neurotrophic factor (BDNF) was done.
RESULTSPretreatment with 200 mg/kg of GLE protected pyramidal neurons in the cornu ammonis 1 (CA1) area from ischemic insult area (F = 29.770, P < 0.05) and significantly inhibited activations of astrocytes (F = 22.959, P < 0.05) and microglia (F = 44.135, P < 0.05) in the ischemic CA1 area. In addition, pretreatment with GLE significantly increased expressions of SOD1 (F = 28.561, P < 0.05) and BDNF (F = 55.298, P < 0.05) in CA1 pyramidal neurons of the sham- and ischemia-operated groups.
CONCLUSIONSOur findings indicate that pretreatment with GLE can protect neurons from ischemic insults, and we suggest that its neuroprotective mechanism may be closely associated with increases of SOD1 and BDNF expressions as well as attenuation of glial activation.
9.Pretreatment of Populus tomentiglandulosa protects hippocampal CA1 pyramidal neurons from ischemia-reperfusion injury in gerbils via increasing SODs expressions and maintaining BDNF and IGF-I expressions.
Tae-Kyeong LEE ; Joon Ha PARK ; Ji Hyeon AHN ; Hyunjung KIM ; Minah SONG ; Jae-Chul LEE ; Jong Dai KIM ; Yong Hwan JEON ; Jung Hoon CHOI ; Choong Hyun LEE ; In Koo HWANG ; Bing-Chun YAN ; Moo-Ho WON ; Il Jun KANG
Chinese Journal of Natural Medicines (English Ed.) 2019;17(6):424-434
To examine the effects of Populus tomentiglandulosa (PT) extract on the expressions of antioxidant enzymes and neurotrophic factors in the cornu ammonis 1 (CA1) region of the hippocampus at 5 min after inducing transient global cerebral ischemia (TGCI) in gerbils, TGCI was induced by occlusion of common carotid arteries for 5 min. Before ischemic surgery, 200 mg·kg PT extract was orally administrated once daily for 7 d. We performed neuronal nuclear antigen immunohistochemistry and Fluoro-Jade B staining. Furthermore, we determined in situ production of superoxide anion radical, expression levels of SOD1 and SOD2 as antioxidant enzymes and brain-derived neurotrophic factor (BDNF) and insulin-like growth factor I (IGF-I) as neurotrophic factors. Pretreatment with 200 mg·kg PT extract prevented neuronal death (loss). Furthermore, pretreatment with 200 mg·kg PT extract significantly inhibited the production of superoxide anion radical, increased expressions of SODs and maintained expressions of BDNF and IGF-I. Such increased expressions of SODs were maintained in the neurons after IRI. In summary, pretreated PT extract can significantly increase levels of SODs and protect the neurons against TGCI, suggesting that PT can be a useful natural agent to protect against TGCI.
Animals
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Brain-Derived Neurotrophic Factor
;
genetics
;
metabolism
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CA1 Region, Hippocampal
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drug effects
;
metabolism
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Gerbillinae
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Humans
;
Insulin-Like Growth Factor I
;
genetics
;
metabolism
;
Male
;
Neuroprotective Agents
;
administration & dosage
;
Plant Extracts
;
administration & dosage
;
Populus
;
chemistry
;
Pyramidal Cells
;
drug effects
;
metabolism
;
Reperfusion Injury
;
drug therapy
;
genetics
;
metabolism
;
Superoxide Dismutase
;
genetics
;
metabolism
;
Up-Regulation
;
drug effects