2.Differentiation of mouse embryonic stem cells into insulin-secreting cells induced by a 5-step model system
Mu-Chao WU ; Hua CHENG ; Ming-Tong XU ; Li-Hong CHEN ; Feng LI ; Sheng-Neng XUE ;
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.
3.The application of dual-source CT in coronary angiography combined with right ventricular function analysis
Qiang LI ; Nong QIAN ; Sheng LI ; Hongzhang TONG ; Liefu XU ; Jianhua WANG
Chinese Journal of Radiological Medicine and Protection 2013;33(4):438-441
Objective To explore the accuracy and radiation dose of the right ventricular analysis with DSCT(dual-source computed tomography)using dual-step prospective ECG trigger.Methods Fortyeight consecutive patients who were suspected or diagnosed with coronary artery disease were examined with DSCT coronary angiography and MRI ventricular analysis.Sequential acquisition and dual-step prospective ECG-trigger were used with 30%-90% width R-R interval.Full tube current output was adopted at 70% (HR ≤70 bpm)or 40% (HR > 70 bpm) R-R interval according to heart rates,while 20% current output was adopted in other R-R interval.Coronary artery was divided into 16 segments according to the American Heart Association.Image quality of coronary arteries were graded with 4-points scale.The DSCT date was reconstructed with 5% R-R interval.RVESV,RVEDV and RVEF were evaluated in DSCT and MRI.Results Forty-two cases accomplished DSCT and MRI examination.In 558 evaluated coronary segments,96.42% could be diagnosed.The average radiation dose was(2.82 ± 0.55)mSy.Paired t-test indicated that the RVESV,RVEDV and RVEF of DSCT and MRI had no statistically significant differences (t =-0.28,0.44 and 1.49,P>0.05),and the correlation was high (r =0.89,0.89,0.87).Conclusions The two generation DSCT with dual-step prospective ECG-triggered sequential acquisition can be used in coronary angiography and right ventricular function analysis simultaneously,which is high in imaging quality of coronary artery,reliable in right ventricular function analysis,as well as lower in radiation dose.
4.Analysis of changes in stray light and relevant factors before and after femtosecond laser in situ keratomileusis
Yang-chen, LIU ; Yan, WANG ; Jing, LI ; Tong, ZUO ; Lu, WANG ; Xi-liu, BAO ; Sheng-sheng, WEI
Chinese Journal of Experimental Ophthalmology 2012;(12):1096-1099
Background As a newly developed refractive surgery,femtosecond laser in situ keratomileusis (LASIK) receives more and more attention.Changes in ocular stray light after femtosecond LASIK is an important problem that impacts visual quality.Some relevant research has been performed before,but the outcomes are conflicting.Objective This primary study was to investigate the change of stray light values before and after femtosecond LASIK and analyze the relevant factors underlying the change.Methods Clinical data from 109 eyes of 55 myopic patients who underwent femtosecond LASIK in Tianjin Eye Hospital from December,2010 through February,2011 were included in this study.Stray light values were measured and recorded by C-Quant stray light meter preoperatively,and 1 day,1 week,1 month and 6 months postoperatively.The stray light data were compared by repeated measures ANOVA,and the correlations among the stray light values with preoperative factors,such as age,equivalent sphere,pupil diameter and central corneal thickness(CCT),as well as with postoperative factors,such as residual bed thickness(RBT),RBT/CCT,ablation depth,ablation ratio,flap thickness,flap diameter flap base width and exiting energy,were analyzed statistically.Written informed consent was obtained from each individual at initial of this study.Results The stray light values were 0.90±0.19,1.10±0.19,1.02±0.18,0.96±0.16 and 0.94±0.15 pre-operation,and 1 day,1 week,1 month and 6 months after femtosecond LASIK.The stray light values increased significantly 1 day,1 week,and 1 month after surgery,showing significant differences among the various time points (F=17.699,P=0.000).The mean stray light value was higher in 1 week than that in 6 months after operation (t=2.412,P=0.017).No significant differences were found in the stray light values between 6 months and 1 month or pre-operation (t =0.779,P =0.437 ; t =-1.877,P =0.062).No significant correlations were seen between preoperative and postoperative stray light values with age,preoperative refractive diopter,pupil diameter or CCT (P>0.05).The changes in stray light values 1 week,1 month and 6 months after surgery were negatively correlated with ablation depth(r=-0.226,-0.228,-0.241 ;P<0.05) and ablation ratio(r =-0.149,-0.219,-0.255 ;P<0.05).Conclusions Stray light values increase within 1 month and return to normal 6 months after femtosecond LASIK.Stray light values may be related to multiple setting laser parameters and factors during femtosecond LASIK.
5.Treatment of non-traumatic femoral head avascular necrosis by perfusion of bone marrow stromal stem cells through optional artery.
Pei-Jian TONG ; Fu-Sheng YE ; Shan-Xing ZHANG ; Ju LI ; Liu XIN-QI
China Journal of Orthopaedics and Traumatology 2014;27(7):565-569
OBJECTIVETo study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis.
METHODSFrom January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis.
RESULTSThe patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening.
CONCLUSIONPerfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.
Adult ; Angiography, Digital Subtraction ; Arthroplasty, Replacement, Hip ; Female ; Femur Head Necrosis ; therapy ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged
6.Effects of Hypericum perforatum L. extract on anti-stress response. In normal mice and antioxidant activity in exhaustive exercise mice.
Ting-Ting YE ; Le LI ; Ying-Ying LIAO ; Ji-Zhong YAN ; Sheng-Qiang TONG
Chinese Journal of Applied Physiology 2014;30(4):332-334
Animals
;
Antioxidants
;
metabolism
;
Fatigue
;
blood
;
Female
;
Hypericum
;
chemistry
;
Male
;
Mice
;
Mice, Inbred ICR
;
Physical Conditioning, Animal
;
Plant Extracts
;
pharmacology
7.Study on biolgical properties of hemopoiesis-supporting function and others of human bone marrow stromal cells
Jicheng YANG ; Weihua SHENG ; Lie LI ; Ningzheng TONG ; Hongwei WANG ; Chun GUO ; Xiaodong WANG
Journal of Cellular and Molecular Immunology 2001;17(1):41-45
Aim To study on biological properties and hemopoiesis-supporting function of human bone marrow stromal cells cultured for long-term in vitro. Methods ① Bone marrow stromal cells from fetus, chidren and adults were cultured for long-time in vitro by static adherent wall assay. ② The phenotypes of the stromal cells were analyzed by immunocytochemical staining or FACA.③ Hemopoietic stem cells in umbilical cord blood were co-cultured and expanded with the stromal cells at different development stages. Results ① The fibroblast myoid cell lines were established and could be propagated to 10 generations and kept for 6 months, meantime, endothelial cells and macrophoges were obtained. ② The child stromal myoid cells were positive for viementin and negative for VIII factor by immunocytochemical staining and their phenotypes were CD33-,CD34-,CD38-and CDW90+ , while the phenotypes of adult stromal cells were CD33-, CD34-, CD38+ and CDW90+ . ③ The stromal cellls could support long-time survival or expansion of LTC-ICs. Survival time of LTC-ICs in the stromal cell-supporting system supplemented with SCF, IL-3 and IL-6 was even longer than that in system without stromal cells(P∨ 0.01). The productivity rate of LTC-ICs cultured in the stromal cell-supporting system was 2 to 4 times higher than that cultured in system without stromal cells. Conclusion The human bone marrow stromal cells could be cultured for long-time in vitro and have hemopoiesis-supp ortingfunction for the hemopoietic stem cells in cord blood.
8.The Study of Treatment of Odor Gas Emitted from the Wastewater Plant by the Biofilter
Jian-Jun LI ; Yan-Zhen LIANG ; Tong-Sheng CHEN ; Ying-Hua CEN ; Guo-Ping SUN ;
Microbiology 1992;0(05):-
A pilot-scale research on purification of odorous gas emitted from wastewater treatment plant using a biofilter was conducted. The aim of this study is to check on the performance of biofilter running in various conditions and the effect of pH fluctuations on the performance of biofilter. The relation between distribution of microorganism and removal of odorous gases were also discussed here. The experimental results show that the predominant odor-causing gas can be efficiently eliminated by a biofilter inoculated with deodoring microorganism which were isolated previously. Moreover the biofilter had been proved having good tolerance to shocking loads of pollutant and can operate well in the condition of low pH.
9.Effects of hypothermia on the liver in a swine model of cardiopulmonary resuscitation
Yi HAN ; Zhen-Ju SONG ; Chao-Yang TONG ; Chun-Sheng LI
World Journal of Emergency Medicine 2013;4(4):298-303
BACKGROUND: The study aimed to explore the effects of hypothermia state induced by 4 oC normal saline (NS) on liver biochemistry, enzymology and morphology after restoration of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in swine. METHODS: After 4 minutes of ventricular fibrillation (VF), standard CPR was carried out. Then the survivors were divided into two groups: low temperature group and normal temperature group. The low temperature (LT) group (n=5) received continuously 4 oC NS at the speed of 1.33 mL/kg per minute for 22 minutes, then at the speed lowering to 10 mL/kg per hour. The normal temperature (NT) group (n=5) received NS with normal room temperature at the same speed of the LT group. Hemodynamic status and oxygen metabolism were monitored and the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured in blood samples obtained at baseline and at 10 minutes, 2 hours and 4 hours after ROSC. At 24 hours after ROSC, the animals were killed and the liver was removed to determine the Na+-K+-ATPase and Ca2+-ATPase enzyme activities and histological changes under a light or electron microscope. RESULTS: Core temperature was decreased in the LT group (P<0.05), while HR, MAP and CPP were not significantly decreased (P>0.05) compared with the NT group (P>0.05). The oxygen extraction ratio was lower in the LT group than in the NT group (P<0.05). The serum levels of ALT, AST and LDH increased in both groups but not significantly in the LT group. The enzyme activity of liver ATP was much higher in the LT group (Na+-K+-ATP enzyme: 8.64±3.32 U vs. 3.28±0.71 U; Ca2+-ATP enzyme: 10.92±2.12 U vs. 2.75±0.78 U, P<0.05). The LT group showed less cellular edema, inflammation and few damaged mitochondria as compared with the NT group. CONCLUSION: These data suggested that infusing 4 oC NS continuously after ROSC could quickly lower the core body temperature, while maintaining a stable hemodynamic state and balancing oxygen metabolism, which protect the liver in terms of biochemistry, enzymology and histology after CPR.
10.Fabrication and optimization of HLA-DRB1-12 oligonucleotide microarray.
Shuang-Ding LI ; Li TONG ; Su-Hong CHENG ; Yu DING ; Sheng-Bin LI ; Sheng-Qi WANG
Journal of Experimental Hematology 2003;11(4):393-397
Oligonucleotide microarray is developed on the basis of hybridization on the solid substrate. The pre-activated glass substrates and the terminal modification of the oligonucleotides are the two important factors in the process of fabrication for microarray. In order to compare the hybridization signal intensity of the different terminal modified oligonucleotide probes, the eight kinds of oligonucleotides were designed according to the sequence of HLA-DRB1-12, including the amino modified oligonucleotides with PEG spacer and the one without spacer, the phosphorothioate modified oligonucleotides with PEG spacer and the one without spacer. They were modified on 5' terminal and 3' terminal, respectively. In addition, the oligonucleotides probes with the internal spacer of different number of PEG were designed to observe the relationship between the spacer of PEG and the hybridization efficiency. These probes were respectively fixed on the bromoacetylation activated and glutaraldehyde activated slides to manufacture the two kinds of microarray which hybridized with the fluorescence labeled PCR product of HLA-DRB1-12 gene. The results from the study demonstrated that the signal intensity of 3' amino-modified probes with the internal spacer of different number of PEG on the bromoacetylation activated slides was stronger than the others. It is concluded that the 3' amino-modified oligonucleotide with an internal PEG spacer and the bromoacetylation activated slide enhanced the hybridization efficiency and were worthy to be proposed for the fabrication of HLA microarray or other kinds of microarrays for detecting fluorescence labeled PCR product in the future study.
HLA-DR Antigens
;
genetics
;
HLA-DRB1 Chains
;
Humans
;
Oligonucleotide Array Sequence Analysis
;
methods