1.Induce and differenctiation of marrow mesenchymal stem cell into photoreceptor-like cell in vitro and microenvironment
Chinese Journal of Experimental Ophthalmology 2013;(7):659-663
Background Marrow mesenchymal stem cells (MSCs) has been successful induced to differentiate into corneal cells,retinal ganglion cells (RGCs) and retinal neuron-like cells in recent years,but there are few studies about MSCs induced into photoreceptor cells and their microenvironment.Objective The aim of this study was to explore the induce and differentiation of bone marrow mesenchymal stem cells (BMSCs) into photoreceptor-like cells in vitro and microenvironment.Methods The second generation of human BMSCs strain and RPE cells strain were cultured and passaged,respectively,and the fourth generation of BMSCs and the third generation of RPE cells were used in the experiment.BMSCs were cocultured using the mesenchymal stem cells medium (MSCM) contained 20 μg/L basic fibroblast growth factor (bFGF),20 μg/L epithelial growth factor (EGF)and 20 μg/L brain derived neurotrophic factor (BDNF) with RPE cells to induce the differentiation of BMSCs in the induced group,and BMSCs were cultured in MSCM only in the control group.The morphology of induced and differentiated cells were observed under the inverted microscope.Inmmunocytochemistry was used in induced for 3-,5-,7-day cells to detect the expression rate of rhodopsin protein for the identification of phenotype of the differentiated cells.RT-PCR was used in induced for 5-,7-day cells to detect the expressions of rhodopsin mRNA and recoverin mRNA.Results Cultured BMSCs grew well with the spindle shape,and passaged RPE cells showed the uniform size and polygon shape with the abundant pigment in the cells.Some induced cells appeared to be the neuron-like cells with round shape and long prominence and the secondary reticular branches.The expression rates of rhodopsinin the cells were (5.83±0.29)%,(20.36±0.32)% and (29.80±2.30)% in the third,fifth and seventh day after induce,which were significantly higher than (0.71 ±0.35) %,(2.56±0.24) % and (2.32±0.42) % of control cells (t3 d =41.510,t5d =107.290,t7 d =30.036,P<0.01).The grey scales of rhodopsin mRNA and recoverin mRNA were significantly elevated in the induced and differentiated cells compared with control cells in the fifth and seventh day (rhodopsin mRNA:t5 d =103.506,t7 d =122.584,P<0.01 ; recoverin mRNA:t5 d =106.674,t7 d =189.992,P<0.01).Conclusions BMSCs can be successfully induced to differentiate into photoreceptor cells after cocultured by conditioned medium with RPE cells.
2.Interaction of Mina and IL-4 gene and their relevance to asthma children
Yun YU ; Qiang XU ; Naizhou GUO
Chinese Journal of Immunology 2014;(5):684-686
Objective:To study the relevance of Mina rs4857304 (G/T) gene locus, IL-4 gene locus rs2243248 (T/G), rs2070874 ( T/C ) single nucleotide polymorphism and allergic asthma pathogenesis of Chinese children , and further analysis of the in-teraction among three loci.Methods:Based on the case-control study, we have analyzed the different distributions , frequencies, inter-action of alleles and genotypes between asthma patients and patients in control group .Results: The Mina rs4857304 sites of patients were compared, and there was statistically significant difference between two groups ( P=0.001, OR =1.773,95%CI =1.256 -2.503);the rs4857304T allelic frequency of patients with asthma was higher than that of the patients in control group , but there was no statistically significant difference about the distribution of different locus genotypes of patients between two groups (P>0.05);MDR interaction analysis have showed that Mina gene loci , rs4857304 and IL-4loci rs2243248, consisting of 2 sites optimal model;Mina loci rs4857304, IL-4loci rs2070874 and rs2243248 that consisting of 3 sites optimal model.There was statistically significant difference be-tween two groups(P<0.05).Conclusion:Allergic asthma would be associated with Mina loci rs4857304, while there might be sig-nificant interactive effects among Mina rs4857304, IL-4loci rs2243248 and rs2070804.
5.Studies on a new experimental system of erythrocyte regulating IL-8 in patients with primary hepatocarcinoma
Lezhi ZHANG ; Feng GUO ; Yu XU
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To approach the capacity of erythrocyte regulating IL-8 in patients with primary hepatocarcinoma (PHC) by a new experimental system of hemaimmune. Methods 0.2ml suspension of cancer cells (S180: 5?10~6/ml) or NS were added into 0.2ml anticoagulant suspension of whole blood cells or leukocytes and 0.3ml plasma, then incubated for 1 h at 37℃. The content of IL-8 was determined by ELISA. Results In the patients with PHC, the IL-8 levels (pg/ml) in experimental and control groups of whole blood cells, and in experimental and control groups of leukocytes were 376.35?243.96, 353.64?271.92, 461.27?277.11 and 424.97?278.93, respectively; while in the normal human, they were 11.36?6.93, 4.98?4.35, 29.41?30.66 and 20.77?24.20, respectively. In the patients with PHC, the activation rates of cancer cells in the experimental groups of both whole blood cell and leukocyte were 0.22?0.24 and 0.25?0.53, respectively; while in normal human, they were 2.49?2.33 and 0.75?0.21, respectively. In the patients with PHC, the IL-8 adsorption rate of erythrocyte in both experimental and control groups of whole blood cell were 0.22?0.18 and 0.17?0.33, respectively; while in normal human, they were 1.18?2.29 and 0.86?0.49, respectively. The IL-8 activated rates of erythrocyte in the patients with PHC was much lower than in the normal human (P
6.Simultaneous Determination of Resveratrol and Polydatin in Polygonum Cuspidatum by Quantitative Nuclear Magnetic Resonance Spectroscopy
Shan YU ; Qiangsheng GUO ; Huilin WANG ; Jianping GAO ; Xu XU
Chinese Journal of Analytical Chemistry 2015;(1):69-74
A quantitative nuclear magnetic resonance spectroscopic ( qNMR) method was established for the simultaneous determination of resveratrol and polydatin in Polygonum Cuspidatum traditional Chinese herb cuts and granule. The 2_step ultrasonic extraction method using 80% alcohol and acetone was used for fully extracting these two components in samples before qNMR determination. The qNMR experimental conditions were investigated and deuterated dimethyl sulphoxide_deuterium oxide (10∶1, V/V) was selected as solvent, the pulse delay time was 5 s, the scan number was 32, 2,3,5_triiodobenzoate was used as internal standard which was calibrated with primary standard substance of potassium hydrogen phthalate. The 1 H_NMR peaks atδ6. 388-6. 391 ( d, 2H) of resveratrol and δ 6. 322-6. 330 ( t, 1H) of polydatin were chosen as the quantitative peaks. Method validation was performed in terms of precision ( RSD<0. 6%), linearity (correlative constants R2>0. 999), limit of detection (0. 23 g/L for resveratrol and 0. 24 g/L for polydatin) and limit of quantitation ( resveratrol 0. 69 g/L, polydatin 1. 57 g/L), recovery ( resveratrol 97. 7% -103 . 5%, RSD=2 . 4%, polydatin 94 . 5%-99 . 2%, RSD=1 . 6%, including the sample extraction and preparaton process) . The results showed the reliability of qNMR for traditional Chinese medicine assay. The resveratrol and polydatin in Polygonum Cuspidatum real cuts and granule samples were experimental determined as 3. 57-5. 69 mg/g and 12. 73-24. 07 mg/g, respectively.
9.Application evaluation of critical value fists data
Dagan YANG ; Xichao GUO ; Genyun XU ; Weimin ZHANG ; Yu CHEN
Chinese Journal of Laboratory Medicine 2008;31(6):695-698
Objective Analyze the historical data of critical values lists,providing scientific evidence for continuous improvement of critical value systems.Methods Screen out critical value lists data of 2006 from laboratory information system,after pretreatment and transformation of data,calculate the percentage of critical value,and its daily distribution,weekly distribution and department distribution, evaluate the range and turnaround time for critical value.Results The rate of critical value was 1.67%.It was mainly concentrated from 8 to 13 O'clock.Monday and Thursday have more critical value than other days.From the perspective of department,the majority critical value was from hematology department and transplantation department.After the evaluation of distribution diagram of critical value range,the lower critical value limit of blood potassium was adjusted from 3.0 mmol/L to 2.8 mmol/L,the blood platelet and leukocyte counts for parlents with hematology disease were a(Ijusted from 20×109/L,1.5×109/L to 10×1O9/L,1.0×109/L respectively.The laboratory turnaround time for 76.2% critical value was less than 1 hour.Conclusion Review and analyze critical value lists data regularly can improve the work efficiency and quality for the laboratory and clinic department and better meet patients' safety needs.
10.Ten years of follow-up study of 27 cases of elderly abdominal aortic aneurysm with ultrasound
Yu PENG ; Fajin GUO ; Guang XU ; Defa CHU
Chinese Journal of Geriatrics 2003;0(09):-
Objective To study the regularity of occurrence and development in the elderly with AAA. Methods Twenty-seven elderly cases with abdominal aortic aneurysm(AAA) were analyzed and followed up with ultrasound for ten years. Results The AAA diameters were 2.5 ~6.6 cm,with the average in crease of(0.14 ? 0.01) cm/ year. Intraluminal thromi(ILT) were found in 63.0% of all the patients. ILT were found(3.9?2.0 ) years after the AAA diagnosis with the average in crease of(0.21 ? 0.05)cm / year. Conclusions The size of AAA in the elderly is slowly enlarged to varying degree with ageing. ILT are found in more than half of all the patients. A few cases of AAA are found ruptured and a few cases of ILT exfoliated.