1.Influence of T- and B-cell-deficiency on retinal neurocytes of mice withacute ocular hypertension
Yanjiao, HUO ; Ping, HUANG ; Shaomin, ZHANG ; Chun, ZHANG
Chinese Ophthalmic Research 2010;28(3):193-197
Background Recently,the study on the cause of optic nerve damage induced by glaucoma is of concern in ophthalmology.Some research showed that the immune system is associated with glaucoma-induced optic neuropathy.Acute ischemia-reperfusion is an ideal model of studying optic neuropathy.ObjectiveThe present study investigates the effect of T and B lymphocyte deficiency on the retinal neurocytes of mice with acute intraocular hypertension.Methods Sixteen SPF CB-17/Icr.Cg-Prkdc~(scid)Lyst~(bg)/CrlVR mice 6-8 week-old (severe combined immunodeficiency mouse,SCID) were used in this study and 16 age-matched SPF wild type (C57BL/6) mice served as controls.The ischemia-reperfusion injury models were induced in the right eyes of 10 SCID mice and 10 C57BL/6 mice through intra-anterior chamber infusion of balanced saline solution for 45minutes to increase the intraocular pressure to 104mmHg,and the left eyes served as model controls.The other 6 SCID mice and 6 C57BL/6 mice served as normal control group.10g/L (2μL) of FlouroGold was injected into the brains of the mice for the labeling of surviving retinal ganglion cells 21 days after ischemia-reperfusion.The thickness of retinal inner nuclear layer was measured by H&E staining under the fluorescent microscope 21 days after ischemic insult.The use of the animals followed the Standard of Association for Research in Vision and Ophthalmology.Results In normal control mice,the morphology of retinal ganglion cells (RGCs) and retinal structure were similar between SCID mice and C57BL/6 mice.The differences in the numbers of RGCs and retinal thickness were insignificant between the two types of mice(P>0.05).In the experimental mice,the surviving RGCs were strikingly increased in SCID mice (91%±5%) compared with C57BL/6 mice(78%±5%)(P=0.003).The thickness of the retinal inner nuclear layer was obviously thinner in the model eyes (22.44±1.70μm) compared to model control eyes (31.06±3.75μm) in C57BL/6 mice(P=0.004),but no statistically significant difference was found between the model eyes and model control eyes in SCID mice (33.52±2.13μm vs 34.06±3.00μm) 21 days after ischemia-reperfusion injury(P>0.05).Conclusion T and B lymphocytes deficient mice show a better tolerance to acute intraocular hypertension than the wild type C57BL/6 mice.
2.Specific induction of anti-leukemia effects by umbilical cord cell-derived CD8+ T cytotoxic lymphocytes.
Xin LIU ; Huo TAN ; Chun-Yan WANG ; Zhen-Qian HUANG ; Huan-Zhu ZHANG
Chinese Journal of Hematology 2006;27(7):452-455
OBJECTIVETo explore the specific anti-leukemia immune response of CD8+ cytotoxic T lymphocyte (CTL) derived from cord blood (CB) ex vivo and evaluate the feasibilities and values of the CTL for specific immunotherapy.
METHODSDendritic cells (DC) were induced from mononuclear cells (MNC) by combination cytokines in 10 CB samples. Loading U937 cell lysate antigen on the mature DC, they could stimulate the lymphocytes of the same origin to generate CTL. MidiMACS was used to isolate CD8+ CTL. Analysis of DC was performed by inverted microscopy, scanning electron microscopy and flow cytometry. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the cytotoxicity of the CTL.
RESULTSCocultured with GM-CSF, IL-4, TNF-alpha and PGE2, CB-MNC could be induced into functional DC with typical morphology. The mean cytotoxicity of CD8+ CTL to U937 cells was significant stronger than that of CD8- CTL and TL at the same E: T ratios. The mean cytotoxicity rate of CD8+ CTL to U937 cells was higher than that to K562 cells [(66.36 +/- 12.43)% vs (41.97 +/- 14.24)%] at E: T ratio of 40: 1 (P < 0.05). The cytotoxicity of CD8- CTL to K562 cells showed no difference from that to U937 cells (P > 0.05).
CONCLUSIONMature CB-DC loading U937 cell antigens could induce CB-T lymphocytes to generate leukemia-specific CD8+ CTL. The cytotoxicity of the CD8+ CTL is specific against U937 cells and is more potent than that of CD8- CTL.
CD8-Positive T-Lymphocytes ; cytology ; immunology ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; immunology ; Humans ; In Vitro Techniques ; K562 Cells ; T-Lymphocytes, Cytotoxic ; immunology ; U937 Cells
3.Involvement of leukotrine B4 receptors in the inflammatory responses and immunological regulation in vitro.
Chun-guang HAN ; Huo-gao HUANG ; Ming HU ; Wen-yan LUO ; Yue GAO ; Qiong WANG ; Yong-xue LIU
Chinese Journal of Applied Physiology 2009;25(2):273-276
AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.
METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.
RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.
CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.
Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance
4.R1239H mutation of CACNA1S gene in a Chinese family with hypokalaemic periodic paralysis.
Qing KE ; Wei-ping WU ; Xiu-hai GUO ; Quan-gang XU ; De-hui HUANG ; Yan-ling MAO ; Chun-nuan HUO
Chinese Journal of Medical Genetics 2006;23(3):272-274
OBJECTIVEMutation screening was performed to a Chinese family with hypokalaemic periodic paraiysis(HOKPP) for locating the corresponding mutations of gene and for specifying the clinical features associated with mutations.
METHODSThe cilnical features of patients from HOKPP family were summurized. Techniques of target exon PCR and direct sequencing were used to screen the mutation in CACNA1S and SCN4A genes in all numbers of the family.
RESULTSTwo patients of the family showed the typical features of HOKPP: the age of disease onset is during the childhood, acetazolamide is effective to patients treated. A heterozygous point mutation 3716 (G>A) causing R1239H was found in exon 30 of CACNA1S gene of the patients, but not found in normal members of the family.
CONCLUSIONThe mutant R1239H in CACNA1S gene exists in Chinese patients with familial hypokalaemic periodic paralysis.
Adolescent ; Adult ; Base Sequence ; Calcium Channels ; genetics ; China ; DNA Mutational Analysis ; Family Health ; Female ; Humans ; Hypokalemic Periodic Paralysis ; genetics ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction
5.Polymorphism K469E of intercellular adhesion molecule-1 gene and restenosis after coronary stenting in Chinese patients.
Zhao-ping LIU ; Yong HUO ; Jian-ping LI ; Yan ZHANG ; Lin XUE ; Chun-yu ZHAO ; Xiu-mei HONG ; Ai-qun HUANG ; Wei GAO
Chinese Medical Journal 2004;117(2):172-175
BACKGROUNDInflammation is a major cause of restenosis after coronary stenting. Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule that plays a key role in the tight adhesion between leukocytes and vascular endothelium. The object of this study was to investigate the association between the K469E polymorphism of the ICAM-1 gene and restenosis after coronary stenting in North Chinese population.
METHODSThe ICAM-1 K469E polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism method in 124 patients who had undergone coronary stenting and coronary angiography at least 3 months earlier. Information on clinical risk factors and procedure-related data were also collected.
RESULTSOf 124 enrolled patients in total, there were 72 cases of in-stent restenosis. The restenosis rate in this population was 58.1%. The frequencies of the three possible genotypes of the ICAM-1 K469E polymorphism were: KK genotype 50.8%, EE genotype 41.9%, and EK genotype 41.9%. Among restenosis patients, the frequency of the KK genotype was 58.3% and the frequency of E allele carriers was 41.7%. Among non-restenosis patients, the frequency of the KK genotype was 40.4%, and the frequency of E allele carriers was 59.6%. The distribution of these two genotype groups between restenosis and non-restenosis patients was significantly different (P = 0.049). Using multivariate logistic regression, the difference between the two groups was more apparent. The odds ratio of KK homozygotes vs E allele carriers was 2.6, with 95% confidence interval 1.2 - 5.8 (P = 0.018). After grading of risk factors, we found that the KK genotype was a stronger predictor of in-stent restenosis in obesity or hyperlipemia patients, with an odds ratio of 9.3 and 3.7, respectively (P < 0.05).
CONCLUSIONIn our study population, KK homozygotes of the ICAM-1 codon 469 mutation had a higher risk of restenosis after coronary stenting, especially in the case of obese or hyperlipemia patients.
Asian Continental Ancestry Group ; genetics ; China ; Codon ; Coronary Restenosis ; genetics ; Female ; Genotype ; Humans ; Hyperlipidemias ; complications ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Middle Aged ; Obesity ; complications ; Polymorphism, Genetic ; Stents
7.Efficacy of different iron fortificants in wheat flour in controlling iron deficiency.
Jian HUANG ; Jing SUN ; Wen-Xian LI ; Li-Juan WANG ; An-Xu WANG ; Jun-Sheng HUO ; Jun-Shi CHEN ; Chun-Ming CHEN
Biomedical and Environmental Sciences 2009;22(2):118-121
OBJECTIVETo observe the different impacts of electrolytic iron, FeSO4, and NaFeEDTA on body iron store of anemic school students.
METHODSFour hundreds anemic students at the age of 11-18 years were divided into four groups. Of which, three consumed different iron fortificants from wheat flour as food vehicle for six months and one consumed non-fortified flour (control). The fortification level of electrolytic iron, FeSO4, and NaFeEDTA was 60 mg Fe/kg, 30 mg Fe/kg, and 20 mg Fe/kg, respectively. Blood samples were collected at 0, 2, 4, and 6 months and hemoglobin (Hb), serum ferritin (SF), and transferrin receptor (TfR) were measured.
RESULTSThe hemoglobin levels in three intervention groups increased, the increments of Hb in the NaFeEDTA group were significantly higher than that in the other groups. SF and TfR levels increased in the tested groups and body iron store in the NaFeEDTA group was higher than that in the other groups. These parameters did not show any significant changes in the control group.
CONCLUSIONNaFeEDTA and FeSO4 fortified wheat flour has positive impacts on iron status in anemic students and NaFeEDTA is more effective than FeSO4, while electrolytic iron is less effective in improving iron store in anemic students.
Adolescent ; Anemia, Iron-Deficiency ; drug therapy ; Child ; Dietary Supplements ; Dose-Response Relationship, Drug ; Edetic Acid ; chemistry ; pharmacology ; Female ; Ferric Compounds ; chemistry ; pharmacology ; Flour ; analysis ; Food, Fortified ; Humans ; Iron ; chemistry ; pharmacology ; Iron, Dietary ; Male ; Nutritional Status ; Triticum
8.The nutrition status of children aged 6-23 months after three months of Wenchuan Earthquake in Beichuan and Lixian, Sichuan Province.
Li-juan WANG ; Jun-sheng HUO ; Jing SUN ; Wen-xian LI ; Cheng-yu HUANG ; Shao-lin LAI ; Ji-fang HU ; Chun-ming CHEN ; Yu-ying WANG
Chinese Journal of Preventive Medicine 2010;44(8):696-700
OBJECTIVEThis project aims to investigate the nutrition status of children aged from 6 to 23 months living in Beichuan and Lixian where were seriously affected by the earthquake in 2008.
METHODSThe height, body weight and hemoglobin concentration were measured for 478 children from Leigu township, Qushan township in Beichuan county and Zagunao township, Xuecheng township and Putou township in Lixian county; the number of group 6 - 11, 12 - 17, 18 - 23 months were 190, 144, 144 respectively. The height for age (HAZ), the body weight for age (WAZ) and the weight for height (WHZ) were calculated respectively. The low weight, stunting, wasting prevalence, anemic rate were analyzed.
RESULTSThe WAZ scores of 6 - 23 months males and females in Beichuan county and Lixian county ranged from -0.97 to -0.13 and from -0.67 to -0.23, HAZ scores ranged from -1.23 to -0.31 and from -1.25 to -0.38, respectively. The WAZ (-0.89 ± 1.16) and HAZ (-1.20 ± 1.60) of 18 - 23 months infants in Beichuan county were significantly different from the WAZ and HAZ of 6 - 11 months (WAZ: -0.32 ± 0.92; HAZ: -0.58 ± 0.98) and 12 - 17 months (WAZ: -0.47 ± 1.00; HAZ: -0.68 ± 1.34) infants (F values were 7.161 and 4.584, respectively; all P values < 0.05). The WAZ (-0.63 ± 1.03) and HAZ (-1.11 ± 1.15) of 18 - 23 months in Lixian county infants were significantly different from the WAZ and HAZ of 6 - 11 months (WAZ: -0.23 ± 0.93; HAZ: -0.51 ± 1.55) infants and 12 - 17 months (WAZ: -0.58 ± 0.52; HAZ: -0.80 ± 1.19) (F values were 3.156, 4.345, all P values < 0.05). The prevalence of low weight infants (18 - 23 months) these two counties were 15.6% (12/77) and 9.1% (6/66), respectively. The prevalence of stunting in Beichuan and Lixian have reached 26% (20/77) and 24.2% (16/66), respectively. The wasting rate of 18 - 23 months infants had reached 9.1% (7/77) in Beichuan. The total anemia prevalence among infants reached 49.6% (123/248) and 78.8% (178/226) in Beichuan and Lixian, respectively. The prevalence of moderate anemia have reached 7.7% (19/248) and 19.9% (45/226), respectively.
CONCLUSIONAfter the earthquake in Beichuan, Lixian, the growth of infants aged 6 to 23 months old was below average. The prevalence of anemia was high. The malnutrition status of infant and young children aged from 18 to 23 months in Beichuan and Lixian was more serious, should be targeted of nutrition interventions.
China ; epidemiology ; Disasters ; Earthquakes ; Female ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Male ; Malnutrition ; epidemiology ; Nutritional Status
9.Clinical application of micro-implant anchorage for treatment of scissors bite on one-side posterior teeth.
Zeng-quan WANG ; Hui-xia ZHOU ; Gui-ling CHEN ; Yi-long AI ; Chun-huo HUANG ; Yu LU ; Kai-hong TANG ; Li-juan DU ; Hua-bin ZENG
West China Journal of Stomatology 2008;26(1):40-43
OBJECTIVETo evaluate the efficiency of micro-implant anchorage (MIA) for posterior teeth intruded and the result of the treatment of scissors bite on one-side posterior teeth.
METHODSThe study included 3 females and 1 male. All the overextruding upper posterior teeth were intruded by the MIA. The micro-implant screws were inserted into the buccal and lingual alveolar hone of the maxillary posterior teeth or the buccal alveolar hone of mandibular posterior teeth. About 0.833 N force was used to intrude the overgrowthing upper posterior teeth, and about 0.559 N force was used to draw buecally the low posterior teeth tilting lingually.
RESULTSThe overextruding upper posterior teeth were intruded 2.0 mm on average, the low posterior teeth tilting lingually were upreared buccally. All the MIA screws kept stable during the treatment, but there was a slight inflammation around the implant screws.
CONCLUSIONMIA could be used as an efficient method to correct scissors bite on one-side posterior teeth with intruding overgrowth upper posterior teeth, or uprearing buccally the tilting low posterior teeth.
Bone Screws ; Dental Occlusion ; Female ; Humans ; Male ; Molar ; Open Bite ; Orthodontic Anchorage Procedures ; Tooth Movement Techniques
10.Subcellular localization and tissues expression profile of hGPCRc: an orphan G protein-coupled receptor.
Guang-Sheng YUAN ; Guang-Tang PAN ; Fang-Ming WU ; Chun-Guang HAN ; Huo-Gao HUANG ; Ming HU ; Li SHENG ; Jing CHEN ; Yong-Xue LIU
Chinese Journal of Biotechnology 2005;21(3):365-369
As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Amino Acid Sequence
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Animals
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CHO Cells
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Cell Membrane
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metabolism
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Cricetinae
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Cricetulus
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Gene Expression Profiling
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Green Fluorescent Proteins
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genetics
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Humans
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Molecular Sequence Data
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Receptors, G-Protein-Coupled
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genetics
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metabolism
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Tissue Distribution
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Transfection