1.Preliminary results of fetal DNA determination in maternal serum by nested PCR
Thuy Thanh Nguyen ; An Trieu Vu ; Anh Duy Nguyen
Journal of Medical Research 2007;47(2):6-10
Background: Prenatal diagnosis has become a standard part of obstetrics care. Genetic diagnoses are established prenatally through the sampling of fetal genetic material. So that the presence of fetal DNA in maternal plasma has led to exciting possibilities of non-invasive prenatal diagnosis. Objectives:In order to provide a reliable non-invasive method for diagnostic of the sex linked disorders. Subjects and method: Fetal gender was determined in 10 pregnant women in which 6 male fetus and 4 female fetus. They are between 34 and 36 weeks of gestation using DNA extracted from 1.6ml of each maternal serum. Maternal serum was put into vacutainer SST before delivery while two pregnant women were implemented by caesarean section at Hanoi Hospital for Obstetricts and Gynecology. Results:The 198bp SRY gene-specific sequence on Y chromosome, 261 bp ATL1 gene specific sequence on X chromosome were amplified in nested PCR. The results were confirmed by examination of newborn child after delivery.The mean level of using DNA extracted from maternal serum was 8.73 \xb1 2.36 ng/\xb5l. The mean level extracted fromperipheral blood was 66.2 \xb1 7.07 ng/\xb5l. Conclusion: The 198 bpSRY specific sequence we detected in 6 serum sample from pregnant women with male fetus. In the remaining cases bearing female foetuses only the 261 bp ATL1 gene sequence was detected. The result was completely concordant with the examination of the newborn child after delivery.
Serum/ immunology
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Prenatal Diagnosis/ methods
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Polymerase Chain Reaction/ methods
3.Study on sensitization and mechanism of CGA-BSA.
Xiaowu HUANG ; Hongbo LIAO ; Ping LIU ; Hui LIN ; Liyan YUAN ; Yuan HU
China Journal of Chinese Materia Medica 2010;35(9):1181-1184
OBJECTIVETo investigate the sensitization and mechanism of artificial antigen of chlorogenic acid (CGA-BSA).
METHODUsing intensive immunization to establish allergy animal model on guinea pig and preparing antiserum and tissue for further test. Using HE staining to observe pathology change of lungs, trachea, liver. Using passive mast cell (PMC) degranulation test to observe the immunogenicity of CGA-BSA and using ELISA to detect IgE and histamine in plasma.
RESULTThere established allergy animal model on guinea pig, which include a increase cell degranulation by a ratio (63.58 +/- 10.23)% in PMC test, increase of specific antibody IgE and increase of histamine in plasma after provocation by ELISA.
CONCLUSIONAllergen CGA-BSA could provoke allergenic response in guinea pig, and the allergic response belongs to type I allergy.
Animals ; Chlorogenic Acid ; immunology ; Drugs, Chinese Herbal ; adverse effects ; Guinea Pigs ; Histamine Release ; Hypersensitivity ; blood ; immunology ; Immunoglobulin E ; blood ; Mast Cells ; immunology ; Random Allocation ; Serum Albumin, Bovine ; immunology
5.Synthesis, identification of artificial antigen of catalpol and preliminary study of immunogenicity.
Zhuang LI ; Wei-ku ZHANG ; Yue ZHANG ; Geng-ni DI ; Hui-hua QU ; Yan ZHAO ; Qing-guo WANG ; Jie-kun XU
China Journal of Chinese Materia Medica 2015;40(7):1287-1290
The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
Animals
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Antibodies
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immunology
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Antigens
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chemistry
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immunology
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Immunoassay
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Iridoid Glucosides
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chemistry
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immunology
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Male
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Medicine, Chinese Traditional
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Mice
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Mice, Inbred BALB C
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Serum Albumin, Bovine
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chemistry
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immunology
6.Acute Serum Sickness Induced Immunologic Injury of the Choroid Plexus; With Particular Reference to the Effect of Prednisolone and the Nature of the Interstitial Cell.
In Joon CHOI ; Sang Ho CHO ; Dong Sik KIM
Yonsei Medical Journal 1974;15(2):115-127
Immune complex deposits have been found in the choroid plexus in patients with systemic lupus erythematosus, and it can be assumed that an immune complex injury to the choroid plexus might be related to the neuropsychiatric disorder seen in patients with SLE. Acute serum sickness was experimentally induced in rabbits by intravenous injection of crystalized BSA. Prednisolone in conventionl dosage was administered to study the immunologic injury of the choroid plexus as well as the mechanisms involved in the prednisolone effect. Light, electron microscopic and immunofluorescent studies were made. The host immunoglobulins(IgG, IgA, IgM) and beta 1 C globulin were demonstrated in the choroid plexus. Histopathological findings included mild to moderate interstitial and perivascular lymphocyte and plasma cell infiltrations and edema. Control animals showed no immune deposits and no histopathologic changes. Electron microscopic findings comparing the immunofluorescent and histopathologic changes were minimal, and showed sparse, vague electron dense deposits particularly in the interstitial spaces, knob-like focal thickening of vascular basement membrane, swelling of endothelial cells, and some accentuation of interstitial cells. The morphologic and functional similarities of the choroid plexus and glomerular basement membrane, the findings in morphologic, electron microscopic and immunofluorescent examinations of the experimental rabbits, along with the observed effects of prednisolone, together with similar reports in the recent literature suggest that immunologic injury of the choroid plexus could be considered as a new disease entity. This immunologic injury might play a significant role in neuropsychiatric disorders in the long standing immune complex deposit diseases. The very interesting finding is the nature and function of the interstitial cell between the endothelial (vascular) and epithelial side basement membranes, and speculation as to whether or not the role of this interstitial cell in choroid plexus injury may be in its possible analogy with glomerular mesangial cells.
Acute Disease
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Animal
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Choroid Plexus/drug effects
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Choroid Plexus/immunology*
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Choroid Plexus/pathology
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Lupus Erythematosus, Systemic/etiology
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Prednisolone/pharmacology*
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Rabbits
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Serum Sickness/chemically induced
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Serum Sickness/complications*
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Serum Sickness/immunology
8.Antigenicity of the soluble egg antigen of Paragonimus westermani.
Suk Il KIM ; Eng Ku KO ; Shin Yong KANG ; Seung Yull CHO
The Korean Journal of Parasitology 1986;24(1):49-54
To evaluate the immature eggs of Paragonimus westermani as a source of diagnostic antigen, about a million eggs which were excreted by 104 adult worms were collected; their saline extract (soluble egg antigen; PwSEA) was prepared. The specific IgG and IgM antibody levels were observed in experimental dog paragonimiasis by micro-ELISA, using PwSEA as well as whole worm extract of 12 week-old P. westermani (PwWWE). The protein composition of the PwSEA was observed by disc-PAGE. The results could be summarized as follows: Specific IgG antibody to PwSEA began to increase on 8 weeks after the experimental infection; it maintained its high level until the observation period of 13 weeks. The levels of IgM antibody to PwSEA, however, did not show any significant change. Specific IgG antibody to PwWWE began to increase earlier from 2 weeks after the infection and continued to increase until the observation period of 13 weeks. Its level was much higher than that to PwSEA. Specific IgM antibody to PwWWE increased temporarily during 2-8 weeks after the infection. By disc-PAGE, PwSEA showed 2 protein bands of very low motility. The bands of PwSEA corresponded to the first and second bands in the electrophoretic pattern of PwWWE of the 12 week-old worms. The above results indicated that the PwSEA induced antibody production in dog paragonimiasis but its antigenicity was weaker than PwWWE to be used as a diagnostic antigen.
parasitology-helminth-trematoda
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immunology
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Paragonimus westermani
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antigen
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enzyme-linked-immunosorbent assay
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serum
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IgG
9.Evaluation of enzyme-linked immunosorbent assay in serological diagnosis of human neurocysticercosis using paired samples of serum and cerebrospinal fluid.
Seung Yull CHO ; Suk Il KIM ; Shin Yong KANG ; Duck Young CHOI ; Jong Sik SUK ; Kil Soo CHOI ; Young Soo HA ; Chin Sang CHUNG ; Ho Jin MYUNG
The Korean Journal of Parasitology 1986;24(1):25-41
The applicability of micro-ELISA was evaluatd in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of 2.5 micro-g/ml. Serum was diluted to 1:100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of 0.18 in both samples. The results were summarized as follows: The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1%; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5% ; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 of 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.
parasitology-helminth-cestoda
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immunology
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Taenia solium
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cysticercus
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enzyme-linked-immunosorbent assay
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serum
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cerebrospinal fluid
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IgG
10.Paper electrophoretic pattern of serum protein fractions in various parasitic diseases.
Won Young CHOI ; Ok Ran LEE ; Won Koo LEE
The Korean Journal of Parasitology 1974;12(2):157-164
Electrophoretic patterns of human blood serum protein were studied in parasitic diseases including 19 cases of clonlrchiasis, 29 cases of clonorchiasis and other helminthiasis and 22 cases of healthy controls in Kim-hae area. The same study was made on 32 cases of ascariasis, 56 cases of trichocephaliasis, 80 cases of ascariasis and trichocephaliasis, 9 cases of amebiasis and 44 cases of healthy controls in the Seoul Juvenile Center. The total serum protein was performed by Folin method and the serum protein fraction test by Durrum's paper electrophoretic method. The results were as follows: In the case of clonolrchiasis, the total serum protein, albumin fraction and A/G ratio decreased and alpha (1) globulin fraction increased twice as much as in the case of healthy controls. In the case of clonorchiasis and other helminthiasis, alpha(1) globulin fractions increased. In the case of ascariasis, albumin fractions increased but alpha(1) globulin fractions decreased. In the case of ascariasis and trichocephaliasis, alpha(2) and beta globulin fractions decreased. In the case of amebiasis, alpha(1) globulin decresed.
parasitology-helminth-trematoda
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electrophoresis-immunology
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serum
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alpha-1 globulin
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alpha-2 globulin
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beta globulin
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protein