2.Directional differentiation of bone marrow mesenchymal stem cells into oligodendrocytes induced by the combination of various neurotrophic factors
Yongtao XU ; Feng LI ; Tie LIU ; Hongbo YOU ; Zhong FANG
Chinese Journal of Tissue Engineering Research 2009;13(14):2793-2796
BACKGROUND: Myelination following axonal regeneration is a key factor affecting the recovery of spinal cord injury. Oligodendrocyte survival directly affects the myelination following axonal regeneration. OBJECTIVE: To investigate the feasibility of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into oligodendrocytes induced by neurotrophic factors. DESIGN, TIME AND SETTING: The cell molecular biology in vitro study was performed at the Laboratory of Department of Orthopaedics, Tongji Hospital from September 2006 to June 2007. MATERIALS: A total of 5 Sprague Dawley rats aged 2-4 weeks, of both gender were selected. Bilateral femur and tibia bone marrow was obtained to harvest BMSCs. METHODS: At passage 4, BMSCs were incubated in serum-free medium, supplemented with N2, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor for 48 hours, and incubated in medium containing 500 ng/mL insulin-like growth factor I and N2 for 3 days. MAIN OUTCOME MEASURES: Morphological changes were observed using an phase contrast microscope. Semiquantitative RT-PCR was utilized to detect specific marker mRNA expression of oligodendrocytes. Using neuron marker anti-microtubule-associated protein, astrocyte marker anti-glial fibrillary acidic protein, oligodendrocyte marker anti-galactocerebroside, anti-myelin basic protein antibody, immunocytochemical staining was performed to detect the positive rate of the differentiation of BMSCs into oligodendrocytes. RESULTS: Morphological changes in BMSCs during the differentiation into oligodendrocytes: After the induction, a majority of BMSCs presented the morphological characteristics of oligodendrocytes. Cytoplasm retraction towards nucleus, cell process extension towards outwards, and strong refraction were found. With the prolongation of time, several cell processes connected and formed a typical net-shape structure. Specific marker mRNA expression of oligodendrocytes: Following induction, specific strap of myelin basic protein mRNA and galactocerebroside mRNA could be detected. Positive rate of oligodendrocytes: During induction, the positive rates of galactocarebroside, myelin basic protein and microtubule-associated protein were 65%, 45% and 10%, respectively. CONCLUSION: The combination of epidermal growth factor, basic fibroblast growth factor and insulin-like growth factor can effectively promote the directional differentiation of BMSCs into oligodendrocytes.
3.Mechanical and biodegradable properties of absorbable hydroxyapatite/poly-DL-lactide screws
Rong FU ; Xiaobo YOU ; Zhendong ZHONG ; Yan LI
Chinese Journal of Tissue Engineering Research 2013;(38):6709-6716
BACKGROUND:Through compounding hydroxyapatite and poly-DL-lactide, the mechanical properties, physical and chemical properties of the implants can be enhanced.
OBJECTIVE:To detect the effect of hydroxyapatite/poly-DL-lactide composite internal fixation screws in canine femoral condyle cancel ous bone fracture repair.
METHODS:Forty-two beagle dogs were operated to bilateral femoral condyle fracture models. The left side was fixed using hydroxyapatite/poly-DL-lactide screws as experimental group;while the right side was fixed with pure poly-DL-lactide screws as control group. After 2, 4, 8, 12, 24, 36 and 48 weeks, conditions of fracture were observed using X-ray, femoral and screw specimens were observed histopathological y, and the bending strength and the average molecular weight were detected. The biological absorption rate, intensity decay rate and biodegradation rate were calculated.
RESULTS AND CONCLUSION:After 2-48 weeks, bilateral fractures were fixed wel and new bone grew wel , but the biological absorption rate of the screws in the experimental group was lower than that in the control group (P ≤0.01). In the first 2, 4 weeks, the bending strength of the experimental screws was higher than that of the control screw (P ≤0.05), but the biodegradable speed was slower in the former one (P ≤ 0.05 or P ≤ 0.01). The pathological changes were similar in the two groups. After 48 weeks, the fractures were healed and bone tissue reconstruction was completed. Compared with the pure poly-DL-lactide screws, the hydroxyapatite/poly-DL-lactide composite internal fixation screws have better fixation effects, biocompatibility, mechanical properties and biodegradability.
4.Relapse after hematopoietic stem cell transplantation: Report of two cases and review of literature
Qing LI ; Yaohui WU ; Zhaodong ZHONG ; Yong YOU ; Ping ZOU
Chinese Journal of Organ Transplantation 2013;(2):75-78
Objective To evaluate the effect of cytokine-induced killer cells (CIKs) as an adoptive immunotherapy option for treatment of leukemia relapse after allo-hematopoietic stem cell transplantation (allo-HSCT).Methods Two cases of infusion of donor CIKs in patients with leukemia relapse after allo-HSCT were retrospectively analyzed.Patient one relapsed 986 days (+986d) after HLA-matched unrelated donor allo-HSCT.Applications of chemotherapy only resulted in short term remission,but allo-CIKs were successfully expanded from the patient's peripheral blood mononuclear cells of donor origin.Totally five cycles of CIKs infusion were infused as an alternative of adoptive immunotherapy.Patient two had recurrent in the + 158d after HLA-matched sibling alloHSCT.At + 204d and + 294d,two cycles of CIKs which were expanded from donor peripheral blood mononuclear cells were infused.Results One cycle of CIKs was given to patient one after the application of chemotherapy to reduce the tumor burden,and the patient successively achieved complete remission.Again after additional four cycles of CIKs infusion,consistent remission was maintained during the following seven months.Patient two who had relapsed disease posttransplantation,achieved cytological complete remission after withdrawal of immunosuppressants and undergoing chemotherapy combined with G-CSF mobilized stem cell infusion.However,at + 187d,the patient suffered from side-effect of acute graft versus host disease and extramedullary infiltration.The symptoms were alleviated markedly after one cycle of CIKs infusion at + 204d.Moreover,the pain disappeared after an additional infusion at + 294d.And up to the present,the bone marrow aspiration showed complete remission while the extramedullary disease vanished.Conclusion The use of CIKs in the treatment of leukemia relapse after allogeneic bone marrow transplantation can be feasible and well tolerated.
5.Statistical analysis of monitoring data regarding urinary iodine level of infants and babies in Ningxia in 2009
Hui-zhong, WU ; Li, LI ; Wen-ning, YOU ; Hai-tao, GAO
Chinese Journal of Endemiology 2010;29(4):420-422
Objective To determine urinary iodine level among babies and infants aged 0-30 months in Ningxia, and to provide scientific evidence for strengthening iodine supplement or decreasing salt iodine level in special population. Methods One thousand four hundred and eighty-three babies and infants were selected randomly from 664 administrative villages of 22 counties (city or district) by two-stage sampling method. Urinary iodine was tested with the arsenic cerium catalysis spectrophotometric method and related influencing factors were analyzed. Results Urinary iodine median was 216.5 μg/L of the whole autonomous region, and the value that was lower than 100 μg/L, accounted for 19.1%(283/1483), 100 - 300 μg/L accounted for 49.3%(731/1483), higher than 300 μg/L, accounted for 31.6% (469/1483). Urinary iodine median was in 130.6 - 328.4 μg/L of all counties, which was higher than 100 μg/L. The urinary iodine median of men(223.2 μg/L) was slightly higher than that(210.2 μg/L) of female, no significant difference was observed(Z = - 1.76, P > 0.05). Urinary iodine level changed little when child was younger than one year old(Z = - 0.624, P > 0.05). Then the value dropped gradually after one year old(χ2 = 13.59, P < 0.05), decreased with age by month, and the proportion of the value smaller than 100 μg/L was increased gradually. Urinary iodine level(257.5 μg/L) of child whose mother had taken iodine oil pills was higher than that (221.2 μg/L) of child whose mother had significant difference was observed(Z = - 2.54, P < 0.05). The urinary iodine level (239.1 μg/L) of child who received breast feeding was higher than that (204.2 μg/L) of child without breast feeding among one year old and younger infants and babies, significant difference was observed (Z = - 2.74, P < 0.05). Conclusions Current iodine level in iodized salt is probably higher than suitable in Ningxia, and the value could be decreased. It is unnecessary to strengthen iodine supplement procedure in special population other than people in Xiji county.
6.Analysis of monitoring results of urinary iodine levels of 5 special groups in Ningxia
Hui-zhong, WU ; Li, LI ; Wen-ning, YOU ; Chuan-tao, MAO
Chinese Journal of Endemiology 2011;30(2):173-175
Objective To investigate the iodine nutritional level of 5 special groups (newly married women,pregnant women, lactating women, babies and infants, and students) in Ningxia, and to provide the basis for formulating prevention and control strategies. Methods Clustering and random sampling method were used. In 2008 and 2009, in the 22 counties investigated, in every county with 9 townships or more, nine townships were randomly selected according to their sub-area positions of east, west, south, north and center; four villages were randomly selected in each chosen township, four people with special needs and 2 infants were randomly selected for urine samples collection in each chosen village. In every county with 9 or less townships, one township was randomly selected respectively in east, west, south, north and center sub-areas; four villages were randomly sampled in each chosen township, eight people with special needs and 3 infants' urine samples were randomly collected in each chosen village. In the 22 counties, one township was randomly selected respectively in east, west,south, north and center sub-areas, one village elementary school was randomly sampled in each chosen township,twenty students aged 8 - 10 were randomly selected to collect their urine samples in each school. The iodine concentration was determined by arsenic-cerium contact method. Results A total of 6894 copied of urine samples from newly married women, pregnant women, lactating women, babies and infants, and students were examined, the urinary iodine medians were 209.3, 187.4, 184.0, 216.5, 216.3 μg/L, respectively. From low to high in the order was lactating women, pregnant women, newly married women, students, babies and infants. The level of urinary iodine of babies and infants, pregnant women and lactating women were appropriate, the one of newly married women and students were higher than appropriate. The proportion of less than 100 μg/L of urinary iodine of the 5 kinds special groups were 11.1% (53/475), 35.4% (308/871), 35.4% (659/1863), 19.1% (283/1483), 8.4%(185/2202), respectively, while the urinary iodine of the pregnant women and lactating women were relatively high. The urinary iodine medians of the 5 special groups were also very different among counties. Conclusions The urinary iodine of the 5 special groups in Ningxia presents obvious differences between populations and regions.Current iodized salt is sufficient to ensure iodine nutrition needs for the 5 special groups. But married women and students have higher levels of iodine nutrition, indicating that the salt iodine concentration of Ningxia residents have cut space, full consideration of the 5 special groups and regional differences should be taken.
7.Effect of rabbit adipose-derived stem cells transfected by adenoviral vector mediated hTGF-?_1 gene on chondrocyte differentiation in vitro
Zhong FANG ; Feng LI ; Hong-Bo YOU ; Wei XIONG ; Guang-Hui LI ;
Chinese Journal of Rheumatology 2001;0(04):-
Objective To investigate the proliferation of rabbit adipose-derived stem cell(ADSCs) transfected by adenoviral vector mediated hTGF-?_1 gene and its chondrocyte differentiation potential.Methods The Ad-hTGF-?_1 plasmid vetor which had the hTGF-?_1 gene was developed and transfected the ADSCs.The experimental group was the hTGF-?_1 transfected group.The cells enclosed by alginate were cultured in com- plete chondrogenie medium(CMM).The morphology of the cells were observed,and RT-PCR was used to measure hTGF-?_1 and collagenⅡexpression,at the same time western blot and immunohistochemistry were applied to detect the expression of collagenⅡin ADSCs before and after transfected with hTGF-?_1 gene. Results The hTGF-?_1 transfected ADSCs became the polygon and it proliferated well.The RT-PCR result of hTGF-?_1 on the transfected group was better than the control after transtected for 7 day and 21 day.The dif- ference between the two groups was significant(P
8.In vitro experiment of allergic reactions induced by traditional Chinese medicine injections.
Rui-xia KANG ; Rong-li YOU ; Lei WANG ; Lei LEI ; Zhong WANG
China Journal of Chinese Materia Medica 2015;40(13):2503-2507
Allergic reactions caused by traditional Chinese medicine injections (TCMIs) become a greatest concern in the clinic application safety. The integral animal evaluation method commonly used in the preclinical evaluation for allergic reactions of TCMIs was not sensitive, specific, quick and objective in observation indexes. Therefore, more researchers have paid attention to the in vitro test method for evaluating allergic reactions induced by TCMIs. Currently, the methods for evaluating allergic reactions induced by TCMIs are mainly targeted at type I allergic reaction and anaphylactic reaction, with only a few in vitro methods for evaluating type II allergic reaction. In this paper, researchers summarized relevant literatures published about evaluation methods for allergic reactions induced by TCMIs in recent years.
Animals
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Complement Activation
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Drug Hypersensitivity
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diagnosis
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etiology
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Humans
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Injections
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Medicine, Chinese Traditional
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adverse effects
9.Application of laser-captured single-cell PCR in clonality analysis of neuroendocrine cells in gastric adenocarcinoma.
Xuan-Li TANG ; Gen-You YAO ; Zhong-Sheng ZHAO ; Xing-Chang REN
Chinese Journal of Pathology 2008;37(3):202-204
Adenocarcinoma
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pathology
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Adult
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Aged
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Female
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Humans
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Lasers
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Male
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Middle Aged
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Neuroendocrine Cells
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pathology
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Polymerase Chain Reaction
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methods
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Stomach Neoplasms
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pathology