2.Large-scale expansion of clinical-grade human adipose-derived stem cells using the extracellular matrix
Yuehan SU ; Chao WEI ; Pinlei LV ; Yun CAO ; Yun QIU ; Qing ZHENG ; Shudong XIAO ; Zheng WANG
Chinese Journal of Tissue Engineering Research 2014;(10):1521-1531
BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.
3.Dynamic Changes of Th1/Th2 Cell Factor in Induced Sputum and Clinical Significance in Children with My-coplasma Pneumoniae Pneumonia
xiao-hua, HAN ; jun, WANG ; li, SU ; yun-xiao, SHANG ; xu-xu, CAI
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To detect levels of IL - 4 and INF - ? in induced sputum dynamically in children with mycnplasma pneamo-niae pneumonia( MPP), and to analyze the function of Th1 /Th2 cell factor immune response in the genesis and development of MPP, so as to evaluate the clinical value of induced sputum method in MPP research. Methods There were 38 cases who were diagnoses as MPP using 3% high osmotic pressure of saline water to ultrasonic atomizing inhalation for inducing sputum. ELISA was used to detect IL-4 and INF-?. Results The content of IL-4 in acute stage was higher than that in convalescence stage in induced sputum of MPP children. Severe stage was higher than mild stage. However, the comparison between acute and convalescence stage didn't have statistics difference in the content of INF-?, neither did the comparison between severe and mild stage. IL- 4/INF- ? in acute stage was higher than that in convalescence stage. Severe stage was higher than mild stage. In convalescence stage, the comparison of INF - ?, IL - 4, IL - 4/INF - ? between the severe and the mild didn' t have statistic significance. Conclusions IL-4 and INF - ? have participated in the monogenesis of MPP. The disequilibria of Th1 /Fh2 is existed in MPP and Th2 reaction predominates. So induced sputum analysis can be a better way to judge the light or heavy press degree of MPP practically, conveniently and sensitively.
4.Clinical characteristics and treatment of severe encephalitis associated with neurogenic pulmonary edema caused by enterovirus 71 in China
Yu-Cai ZHANG ; Xing-Wang LI ; Xiao-Dong ZHU ; Su-Yun QIAN ; Yun-Xiao SHANG ; Bi-Ru LI ; Xiao-Lin LIU
World Journal of Emergency Medicine 2010;1(2):108-113
BACKGROUND:Hand-foot-mouth disease has become a major public health issue in children in China. In the present prospective study we investigated the clinical characteristics and emergency management of children with severe encephalitis associated with NPE caused by enterovirus 71. METHODS:The study was conducted in 2 pediatric intensive care units (PICUs) over a 2-month period. Clinical records were reviewed of critically ill children with severe encephalitis associated with NPE caused by EV71 who were admitted to PICUs during the period of May to June 2008 in Fuyang. RESULTS:We reviewed the complete records of 36 children, of whom 23 (63.9%) were male and 13 (36.1%) female. Their age ranged from 4 to 48 months, with an average of 15.8 months. Al children except one were under 3 years of age. The overal mortality in these children was 19.4%. The average duration of critical life threatening signs and symptoms was 2.1 days (12 hours-5 days). Nervous system diseases included brainstem encephalitis in 27 children (75%), brainstem encephalitis associated with myelitis in 6 children (16.7%), and general encephalitis in 3 chidren (8.3%), respectively. In 12 patients of NPE (33.3%) pink or bloody bubble sputum and asymmetric pulmonary edema or hemorrhage was the primary manifestation but no typical exanthema was observed. Five children died of acute onset of NPE and / or pulmonary hemorrhage with rapid progression of cardiopulmonary failure within hours after admission. Therapeutic management consisted of mechanical ventilation and administration of mannitol, methylprednisolone, intravenous immunoglobulin (IVIG) and vasoactive drugs, associated with the need of fluid volume resuscitation in 9 (25%) of the 36 children. CONCLUSIONS:In children less than 3 years of age found to be affected by severe EV71 encephalitis associated with NPE, one fifth may die. The major organ systems infected by severe EV71 include the central nervous system, the respiratory system, and the cardiovascular system. Early diagnosis and evaluation, respiratory support, treatment of intracranial hypertension, and mainttenance of function of the cardiovascular system are the most important therapeutic measures.
5.Formula method of medicated diet based on medicinal property combination patterns.
Li MA ; Su-Rong YAN ; Xiao-He LI ; Ou TAO ; Yun WANG ; Yan-Jiang QIAO
China Journal of Chinese Materia Medica 2014;39(13):2392-2395
To propose a formula method of medicated diet based on medicinal property combination patterns in this paper under the context of lack of innovation in medicated diets. By analyzing the property combination patterns of traditional Chinese medicine and commonly used foods recorded in the pharmacopoeia, medicated diet formulae were optimized by using the greedy algorithm, with the property combination patterns of classical formulae based on the syndrome differentiation and treatment. In this paper, the Baihu Rensheng decoction, which is a classical formula for treating lung and stomach heat-derived diabetes, was taken for example in the formula design. As a result, totally 18 medicated diet formulae were developed and proved to be rational in the analysis on traditional Chinese medicines and nutriology. This method expands the way of thinking for personalized diet therapies and provides theoretical basis the industrial development and clinical application of medicated diets.
Animals
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Diabetes Mellitus
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diet therapy
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drug therapy
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metabolism
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Diet
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Diet Therapy
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Drugs, Chinese Herbal
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chemistry
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therapeutic use
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Humans
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Phytotherapy
6.Perioperative fluid management during pediatric kidney transplantation
Peijiao LIAO ; Cuiling SU ; Yun XIAO ; Xiaoling LIANG ; Lijuan HUANG ; Xian REN
Modern Clinical Nursing 2013;(8):35-37
Objective To explore the strategies and effectiveness of perioperative fluid administration during pediatric kidney transplantation.Methods In our study,we evaluated the heart function and compliance of pediatric patients preoperatively,and then formulated perioperative fluid management strategy according to children’s circulation feature.The electrolytic balance was kept and complications were prevented.Results Twenty-nine out of all 48 children experienced electrolyte disturbance to different extents on day 1 postoperatively,which improved on day 2 after prompt management.There were five cases of oliguria,three cases of delayed graft function(DGF),one case of heart failure and multiple organ failure respectively.Conclusion Such nursing strategies like formulation of reasonable fluid infusion plan according to the physiological features of pediatric recipients,keeping the electrolytic balance and careful management of fluid at polyuria,oliguria and anuria,care critical for the children to live through the perioperative periods.
7.Investigation of clonorchiasis in Huachuan Country, Heilongjiang Province
Su, HAN ; Tao, GE ; Xiao-li, ZHANG ; Yun-xia, DONG ; Hong, LING ; Feng-min, ZHANG
Chinese Journal of Endemiology 2013;32(6):651-653
Objective To investigate the prevalence of clonorchiasis among residents of Huachuan Country,Heilongjiang Province and to provide a basis for development of control strategies.Methods From 2011 to 2012,cluster random sampling was performed to survey the incidence of clonorchiasis in Huachuan Country.Fecal specimens were collected and examined the clonorchis sinensis eggs by Kato-Katz method.A questionnaire survey was conducted to collect related information such as age,gender,occupation and eating habits.The infection characteristic was analyzed.Results Totally 884 patients with clonorchiasis were found among 2248 residents,and the infection rate was 39.32%.The infection rate in male[47.15%(611/1296)] was significantly higher than females [28.68%(273/952),x2 =34.55,P < 0.01].The infection rate increased with age,which was higher in the 20-69 years old people,with the highest infection rate in the 50-59 years old groups[45.34% (219/483)].Of the occupational distribution,farmers had the highest infection rate [47.24% (420/889)],followed by cadres and staffs[38.38%(190/495)].Of residents with fresh fish eating history,the prevalence of clonorchiasis was 53.38%(150/281).Conclusions The prevalence of clonorchiasis is still high in Huachuan County.To reduce the prevalence of clonorchiasis,comprehensive prevention measures,health education and group chemotherapy should be carried out.
8.Serum level of anti-monomer C-reactive protein antibody in systemic lupus erythematosus
Wei ZHAO ; Yin SU ; Ru-Lin JIA ; Xiao-Yun SUN ; Zhan-Guo LI ;
Chinese Journal of General Practitioners 2005;0(09):-
Objective To study clinical significance of anti-momoner C-reactive protein (anti- mCRP) antibody in systemic lupus erythematosus (SLE) and assess the relationship between serum CRP and anti-mCRP antibody.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to determine serum level of anti-mCRP antibody in 113 pateints with SLE,65 patients with other rheumatic diseases,including primary Sjgren syndrome,rheumatoid arthritis,osteoarthritis,ankylosing spondylitis and systemic sclerosis,and 32 healthy controls.Serum level of CRP was evaluated by turbidimetry.Clinical manifestations and laboratory indicators of the patients were all recorded.Results Serum level of anti- mCRP antibody in SLE patients was significantly higher than that in patients with other rheumatic diseases and healthy controls,respectively (t=2.502 and 5.352,respectively,P 0.05).Titer of anti-mCRP antibody closely correlated with systemic lupus erythematosus disease activity index score (r=0.248,P0.05). Conclusions Level of Anti-mCRP antibody increased significantly in patients with SLE,which associated with disease activity of SLE and can be used as a valuable marker in evaluating activity of SLE.
9.Generation of Transgenic Mice with Cardiac-specific Overexpressing Heat Shock Protein 27
Li LIU ; Xiao-Jin ZHANG ; Su-Rong JIANG ; Yun-Lin CHENG ;
China Biotechnology 2006;0(11):-
Objective:Increased reactive oxygen species (ROS) formation and by which in turn promotes cardiomyocytes apoptosis is associated with the pathogenesis and progression of various cardiac diseases. Small heat shock protein 27(Hsp27) could protect different cells from oxidative damage. By using tissue nonspecific overexpression Hsp 27 transgenic model, other investigators demonstrated that Hsp27 suppressed successfully kainate-induced seizures and hippocampal cell death in intact transgenic mice, and attenuated mimic ischemia/reperfusion injury in Langendorff-perfused isolated mice heart. As there are complicated and long distance neuro-humoral regulation associated with the development of cardiac diseases, it is better to choose a cardiac-specific overexpression transgenic model to study the effects of Hsp27 in hearts in vivo.Methods:A cDNA encoding human Hsp27 was subcloned into pBSII-SK+ containing the ?-myosin heavy chain (?-MHC) promoter (generously provided by Dr. J. Robbins, Children’s Hospital of Cincinnati, Ohio). BamHI-digested linear transgene consistent with the ?-MHC promoter, Hsp27 cDNA, and poly (A) of human growth hormone (hGH) was microinjected into the fertilized eggs from CBA/BL6 mice. Mice containing the transgene were identified by polymerase chain reaction. Founders revealed by this screening were used to establish independent transgenic lines. Following successful transmission, a range of tissues including heart, lung, liver, brain, skeleton muscle, spleen and kidney was screened by Western blot to confirm the cardiac specific expression of the transgene. Results and Discussion:Transgenic mice expressed Hsp27 under the control of a-MHC promoter. Cardiac tissues from independent TG line expressed abundant Hsp27, and as expected, Hsp27 expressed in cardiac tissues only, whereas none in liver, spleen, lung, kidney, brain and skeleton muscle. Surprisingly, Hsp25, the endogenous isoform of Hsp27 in murine, was downregulated by Hsp27 overexpression (data not shown), which is in contrast to the result of Hollander’s [1]. It needs to determine in future whether Hsp27 expression profile in heart could exert any effect on the regulation of Hsp25.Conclusion: The TG mice overexpression human Hsp27 specifically in the heart by using ?MHC- promotor were created. All TG mice expressed Hsp27 abundantly and cardiac-specifically. To our knowledge, it is the first report of creation of transgenic mice which overexpressing Hsp27 cardiac specifically. This current model suggests that the Hsp27 cardiac-specific over-expression of transgenic mouse remains a robust genetic tool for dissecting molecular and genetic events involving Hsp27, which could be a therapeutic target in heart failure.
10.Screening of Phytase-producing Strain and Cloning of Phytase Gene from the Selected Strain
Yan WANG ; Xiao-Rong GAO ; Qiao SU ; Jing-Yun WANG ; Li-Jia AN ;
Microbiology 1992;0(04):-
A strain with relative higher phytase-producing ability, Aspergillus fumigatus WY-2 was screened from soil. The optimal pH and temperature for activity of the phytase from A.fumigatus WY-2 were 5.5 and 55 ℃, respectively. The gene encoding the phytase was amplified from genomic DNA of the strain by PCR, and a 1.5 kb DNA fragment was obtained and then was cloned into vector pMD18-T. The sequencing analysis revealed that the DNA fragment contained a whole open reading frame (ORF) of phytase gene. The phytase gene was 1459 bp in length included with a 61 bp intron and encoded 465 amino acids. A signal peptide encoding 26 amino acids was found at 5′end of the gene. There were 7 potential glycosylation sites in the phytase. The present phytase showed 91% identity in nucleotide sequence and 91% identity in deduced amino acids sequence to the previously reported A.fumigatus ATCC34625 phytase.