1.Effect of different decellularizing methods on cell-secreted extracellular matrix in vitro
Limei FAN ; Rong XIA ; Xiaochen DOU
Chinese Journal of Tissue Engineering Research 2014;(30):4863-4867
BACKGROUND:Compared with single or composite biomolecular materials, decellulazied matrices are more biomimetic to natural ectocytic surroundings. So cel-secreted extracellular matrix is paid more and more attention in the field of tissue engineering, and the composition of these matrices are influenced by decellulezired preparation methods more or less. But there are few studies about the biological effect of different decellulazried methods on the extracellular matrix. OBJECTIVE:To investigate the composition of cel-secreted extracellular matrices in vitro by different decellularizing methods and their effects as surface modification on cytobiological reaction. METHODS:After the treatment of different decellularizing methods (freeze/thaw cycles, trypsin, weak alkal, detergents), the extracellular matrix was obtained and grouped into four kinds. The biological composition of the extracellular matrix was determined by ELISA assay. Then osteoblasts were seeded onto the four kinds of extracellular matrix surfaces. cells cultured normal y served as controls. The effect of extracellular matrix coatings on cellgrowth and differentiation were determined by MTT test and alkaline phosphatase activity test. RESULTS AND CONCLUSION:The residual components were the most in the freeze-thaw group, fol owed by the detergent and weak alkal groups, and the least in the trypsin group. Compared with the control group, the absorbance value of cells were lower in the freeze-thaw and detergent groups at days 3, 5, 7 after inoculation (both P<0.05);the alkaline phosphatase activity was lower in the trypsin group at days 5, 7 after inoculation (P<0.05), in the weak alkal group at 7 days after inoculation (P<0.05), and in the detergent group at days 3, 5, 7 after inoculation (P<0.05). Therefore, we can harvest more extracellular matrices by the freeze-thaw method, and the extracellular matrix coating synthesized by the freeze-thaw method is more helpful for cellgrowth than others.
2.Repair of cancellous bone defects with compound of allogenic bone marrow mesenchymal stem cells and fibrin
Qiangbing DOU ; Rong KONG ; Liulong ZHU
Orthopedic Journal of China 2006;0(10):-
[Objective]To discuss bone marrow derived mesenchymal stem cells(MSCs) culture in vitro and the curative effect of repairing cancellous bone defects in rabbits with the compound of allogenic MSCs and fibrin.[Method]MSCs were isolated and culture-expanted from rabbit bone marrow.Cancellous bone defects were created on bilateral femur condylar in 15 Japanese big ear rabbits,the left bone defects were treated with allogenic MSCs and fibrin,the right were treated with fibrin only.The bone defects were examined radiologically and histoloyically at the 2nd,5th,8th week after operation.[Result]The defects with only fibrin implantation couldn't heal spontaneously and no bone formation were found at any time-point.In the defects with compound of MSCs and fibrin implantation,there were a few bone formation within 2 weeks.By the 5th week plentiful callus were found,at the end of the 8th week,the defects healed.There was no immune respone except a few inflammatory cells infiltrate.[Conclusion]The compound grafts of allogenic MSCs and fibrin shows low immunogenicity within 8 weeks and is an effective method for repairing cancellous bone defects.
3.ROLE OF GRANULOCYTE COLONY-STIMULATING FACTOR IN REGULATING DENDRITIC CELL SUBSETS AND ITS EFFECT ON IMMUNE RESPONSE
Jingshi ZHOU ; Kefeng DOU ; Rong FU
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To study the role of granulocyte colony-stimulating factor (G-CSF) in regulating dendritic cells (DC) subsets and its effect on immune response. Methods BALB/c mice received 10?g of G-CSF once a day by subcutaneous injection for 6 days. Then, DC were separated from the peripheral blood. The ratio of DC1(CD11c +CD8a -) and DC2(CD11c +CD8a +) and their absolute numbers were determined with flow cytometry. One week after C57BL/6 PBMC stimulation, mixed lymphocyte culture was used to determine the ability of DC to proliferate priming T cells and the intensity of mixed lymphocyte reaction (MLR). Results After stimulation with different doses of G-CSF for 6 days, the absolute number of DC2 was increased from 9.6?10 6/L to 45.4?9.9?10 6/L (P
4.A study on loneliness among people who lost their children in disaster area of 5.12 Earthquake
Xiaolin HU ; Xiaolin LI ; Rong LI ; Xinman DOU
Journal of Central South University(Medical Sciences) 2014;(12):1279-1284
Objective: To evaluate the status of loneliness, the influential factors and the relationship between loneliness and family function among bereaved parents in disaster area of 5.12 Earthquake and to provide basis for mental and family reconstruction for those people suffering from the Earthquake. Methods: A total 190 bereaved parents in disaster area of 5.12 Earthquake were enrolled for an across-sectional study. A face-to-face survey was conducted by following scales: General Data Scale, University of California at Los Angels Loneliness Scale, Emotional and Social Loneliness Scale and Family APGAR Index. Results: hTe score of loneliness ranged from 24 to 69 (mean=43.58, SD=9.61), the score of emotional loneliness ranged from 5 to 24 (mean=11.95, SD=3.60) while the social loneliness ranged from 6 to 24 (mean=13.99, SD=3.69). hTe scores of loneliness, social loneliness and emotional loneliness were all increased among bereaved parents compared to people without loss of their child. The results of multivariate analysis showed that family function and status of fertility significantly impacted the loneliness (P<0.05). Furthermore, there was negative correlation between family function and the loneliness, social loneliness and emotional loneliness (P<0.05). Conclusion: The bereaved parents in disaster area of 5.12 Earthquake showed higher level of loneliness. The better of family function indicated the less of loneliness in bereaved parents. Measures should be taken to promote mental and family reconstruction for such people, including providing effective skills to improve emotional communication and family function and offering technical support for having another baby.
5.Immunoblot analysis of hepatitis C antibody-weak positive samples detected by chemiluminescent assay
Yaling DOU ; Rong RONG ; Xiuzhi GUO ; Jianguo ZHANG ; Anping NI ; Weijuan YAN
Chinese Journal of Laboratory Medicine 2009;32(9):984-988
ted by the ECI analyzer.
6.NEUROPROTECTIVE EFFECT OF c-fos ANTISENSE OLIGODEOXYNUCLEOTIDE UPON APOPTOSIS INDUCED BY SODIUM SELENITE IN CULTURED CORTICAL NEURONS: POSSIBLE CASCADE OF ACTIVATION OF RELATED GENES
Rong XIAO ; Yan DOU ; Jiahui ZHAO ; Rui WANG ; Xiuzhen YAN ; Jiantian QIAO
Chinese Journal of Neuroanatomy 2002;18(2):93-100
To investigate the role of immediate-early gene c-fos in sodium selenite-induced apoptosis and its position in a possible cascade of apoptogenic genes, we compared the time-courses of expression for 5 related genes, including c-fos, during the apop- tosis induced by sodium selenite with or without blockage of c-fos expression by adding c-fos antisense oligodeoxynucleotide ( ASO) in cultured cortical neurons. The results showed that: (1) in control experiments without c-fos ASO adding, 0. 5 μmol/ L sodium selenite-induced apoptosis as revealed by electrophoretic and flow cytometric examinations; at the same time, sodium selenite also induced down-regulation of bcl-2 mRNA expression and up-regulations of mRNAs related to bax, c-fos, p53, and acetylcholinesterase (AChE) genes; (2) in similar experimental conditions with c-fos ASO cotreatment, the sodium selenite-in- duced apoptosis was blocked with the up-regulation of p53 expression still emerging as before, while the changes in expressions of bcl-2, bax, AChE genes were reversed at the same time. The results suggest that c-fos ASO could play a protective role upon cortical neurons from suffering apoptosis induced by sodium selenite, and there might exist a cascade of gene expressions with p53 and c-fos genes being regulated upstream and then bcl-2, bax, and AChE genes being regulated downstream.
7.Cerebral palsy in children with prematurity and its comorbidities
Dianrong SUN ; Mei HOU ; Wenyan LI ; Tanfeng DOU ; Rong YU ; Ke WANG
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(12):911-914
Objective To analyze the comorbidities and gross motor function classifications (GMFCs) of premature children with cerebral palsy (CP) in terms of neurological subtype and gestational age in search of some relationship. Methods Children with cerebral palsy treated at the Qingdao Children's Hospital from 2006 to mid2009 received intelligence capacity tests, ophthalmological consultations, language/speech tests, brainstem auditory evoked potential measurements, electroencephalograms and GMFC evaluations. All of the children were stratified according to neurological subtype, gestational age, comorbidities and gross motor function classification. Results Of all 258 children, spastic diplegic cerebral palsy predominated (183 case, 70.9% ). 124 cases (48.1% ) had visual disorders, 121 (46.9%) had language or speech disorders and 103 (39.9%) showed mental retardation. The frequencies of individual comorbidities were distributed disproportionately between the different neurologic subtypes.GMFC levels also differed with the different CP types. The GMFC levels of diplegics were significantly better than those of the other types. The distribution of comorbidities such as visual disorders, language or speech disorders, and mental retardation was not related to gestational age or type of CP. Conclusions There is some correlation between the neurological subtype, comorbidities and the GMFC levels. But there is no significant correlation between gestation age and the severity of CP.
8.Absorbable bandage wrapping in treatment of severe blast liver injury: a miniature swine model.
Can-Rong LU ; Lin CHEN ; Wen-Bin CHEN ; Chun-Qing DOU ; Rong LIU ; Zhi-Qiang HUANG
Chinese Medical Journal 2011;124(22):3757-3761
BACKGROUNDThe aim of this research was to evaluate the feasibility and efficacy of absorbable bandage wrapping in the treatment of cases of severe liver trauma.
METHODSElectric firecrackers were detonated in 16 miniature swine to produce a severe blast liver injury. After fluid resuscitation, the animals were randomly divided into two groups (n = 8 each) and were either treated with absorbable bandage wrapping of the injured lobe of liver (Group B) or hepatic lobectomy (Group H). Time to hemostasis, blood loss during the treatment period, and other parameters were compared, including postoperative serum total bilirubin (TB), alanine aminotransferase (ALT), and aspartate aminotransferase (AST).
RESULTSBlood loss during the treatment period was significantly lower in Group B than that in Group H ((81.3 ± 26.0) ml vs. (130.8 ± 29.5) ml, P = 0.0031). Serum AST and ALT were transiently increased post-surgically. These transient increases were significantly higher in Group B. No difference in time to hemostasis was noted ((8.70 ± 2.27) minutes vs. (10.28 ± 1.93) minutes, P = 0.1559) in Groups B and H, respectively. Two pigs were humanely euthanized 28 days post-surgically and the wrapped liver lobes appeared atrophies. Microscopically, there was evidence of emerging and mature fibrous tissue.
CONCLUSIONAbsorbable bandage wrapping is both feasible and effective in the treatment of severe blast liver injury.
Animals ; Bandages ; Female ; Liver ; injuries ; surgery ; Male ; Swine ; Swine, Miniature
9.Effects of chitosan on cell proliferation and collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes.
Chang-Suo XIA ; Guang-Xiang HONG ; Rong-Rong DOU ; Xuan-Ying YANG
Chinese Journal of Traumatology 2005;8(6):369-374
OBJECTIVETo study the proliferation and collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes; and the effects of chitosan on cell proliferation and collagen production in the 3 cell types of rabbit flexor tendon.
METHODSThree cell lines of tendon sheath, epitenon, and endotenon were isolated from rabbit flexor tendon and cultured. Cell culture media was added with chitosan. The cell number and production of collagens I, II, and III were measured and compared with those cultured without chitosan. The expression of type I collagen in tendon sheath fibroblasts was determined by quantitative analysis of reverse-transcription polymerase chain reaction.
RESULTSAll 3 cell lines produced collagens I, II, and III. Adding chitosan to cell media resulted in a significant decrease in cell number in all 3 cell lines. In addition, there was a significant decrease in collagens I, II, and III production in all 3 cell lines as well as the expression levels of type I collagen in tendon sheath fibroblasts (P<0.05).
CONCLUSIONSChitosan can inhibit cell proliferation and collagen production of the tendon sheath, epitenon, and endotenon, and may provide a promising approach to obviating tendon adhesion formation clinically.
10.Hereditary hemorrhagic telangiectasia: a report of two cases.
Yan HUANG ; Chen-Tao LIU ; Xiang-Rong ZHENG ; Bo DOU ; Rong HUANG
Chinese Journal of Contemporary Pediatrics 2020;22(9):1041-1042
This article reports two children with hereditary hemorrhagic telangiectasia (HHT). Patient 1 was a boy aged 12 years and was admitted due to intermittent cough and wheezing for more than 10 years. This boy and his mother and grandmother had a history of epistaxis. The boy had a history of the rupture of cerebral arteriovenous malformations. Gene detection showed a heterozygous mutation, c.277C>T(p.Arg93*), in the ENG gene. Patient 2 was a girl aged 13 years and was admitted due to cyanosis of lips for more than 1 year. The girl had a history of recurrent epistaxis and the manifestations of severe decline in pulmonary diffuse function, pulmonary hypertension, dilation of blood vessels at the distal end of lungs, and small arteriovenous communications in both lungs. Children with HHT often lack typical respiratory symptoms, which may lead to missed diagnosis and misdiagnosis in the early stage. Pulmonary computed tomography or right cardiac acoustic contrast can help with the diagnosis of HHT, and gene detection can improve the early diagnostic rate of this disease.
Adolescent
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Child
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Female
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Humans
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Lung
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Male
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Mutation
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Telangiectasia, Hereditary Hemorrhagic
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Tomography, X-Ray Computed