2.Primary culture of adipose-derived stem cells and differentiation induction into myoblasts
xiao-fei, SONG ; qiang, FU ; yue-min, XU ; lei, CUI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(05):-
Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cells in induction group and control group,respectively.The induction lasted for 7 d,14 d,21 d,28 d and 35 d,respectively.Cell growth and cell morphology were observed by inverted phase contrast microscope,and immunofluorescence and flow cytometry were utilized to detect the expression of myoblast specific antigens desmin and myosin. Results ADSCs were successfully isolated and cultured,and were identified to be stem cells.On the 28th day of induction,cells in induction group displayed "swirl" morpholgy,and multinucleation was observed.It was revealed by immunofluorescence and flow cytometry that the highest expression rates of desmin and myosin were 52.57% and 50.04%,respectively on the 28th day of induction,while there was no expression before induction and in control group. Conclusion ADSCs can be isolated and cultured from rat adipose tissues,and can further differentiate into myoblasts after induction by culture media with 5-aza.The expression of myoblast specific antigen is the highest on the 28th day of induction.
3.The distribution of platelet glycoprotein I b? variable number tande repeat polymorphism and the relationship with cerebral infarction
Xiao-Min XIN ; Yu ZHAO ; Lan-Ying CUI ;
Chinese Journal of Laboratory Medicine 2001;0(02):-
Objective To study the distribution of variable number tande repeat(VNTR) polymorphisms of the platelet membrane glycoprotein Ⅰ b? in Han nationality at Harbin and the relationship between these polymorpbisms and cerebral infarction(CI).Methods The identification of alleles and genotypes of VNTR polymorphism of the glycoprotein Ⅰ b? gene was performed by polymerase chain reaction (PCR)in 200 healthy individuls and 200 CI patients(77 lacunar infarction patients and 123 atherosclerotic thrombotic infarction patients),to analyze The relationship between gene polymorphisms and cerebral infarction.Results(1)There were three types of alleles:B、C、D,and five types genotypes:BC,BD,CC, CD,DD in Harbin Han nationality.No person with A allele and BB genotype was found.(2)No statistically significant differences of GP Ⅰ b? gene VNTR polymorphism was found between CI patients or subtype CI patients and controls(P=0.412 and 0.572,respectively).Conclusions(1)This study indicates that the C and D alleles of VNTR polymorphisms of GP Ⅰ b? are the main alleles while the CC and CD genotypes are the main genotypes in Harbin Han people.(2)Our findings indicate that no association exists between the VNTR polymorphism of platelet GP Ⅰ b? gene and CI.
4.Expression of plasma microRNA-214 in patients with acute myocardial infarction and its correlation with left ventricular remodeling
Yali XIAO ; Min ZHAO ; Ping LYU ; Ruijuan LIU ; Jinyan WANG ; Tao CUI
Tianjin Medical Journal 2017;45(5):501-505
Objective To investigate the expression of plasma microRNA-214 in patients with acute myocardial infarction (AMI) and its correlation with left ventricular remodeling (LVR). Methods A total of 158 AMI patients and 85 controls were selected from Hebei province Zhuozhou City Hospital. According to the left ventricular remodeling after PCI operation, AMI patients were divided into LVR group (n=41) and non LVR group (n=105). Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the plasma microRNA-214 levels, and the brain natriuretic peptide (BNP) and C-reactive protein (CRP) levels were detected by immunoassay. Pearson correlation analysis of plasma microRNA-214 levels and the correlation between CRP and BNP were analyzed in patients with LVR. The receiver operating characteristic (ROC) curve analysis of plasma microRNA-214, BNP and CRP were used to predict the efficacy of LVR in patients with AMI after PCI. Logistic regression model was used to analyze the relationship between the above indexes and LVR. Results Compared with the control group, left ventricular ejection fraction (LVEF) and left ventricular short axis shortening (FS) were significantly decreased in LVR group and non LVR group, and which were decreased more significantly in LVR group compared with those of non LVR group (P<0.05). The left ventricular end diastolic diameter (LVDD), left ventricular end diastolic volume (LVEDV) and left ventricular end systolic volume (LVESV) were significantly increased in LVR group and non LVR group, and which were increased more significantly in LVR group than those in non LVR group (P<0.05). The levels of plasma microRNA-214, BNP and CRP were significantly higher in LVR group than those in non LVR group and control group (P<0.05). Correlation analysis showed that microRNA-214, BNP and CRP were positively correlated in patients with LVR (r=0.684 and r=0.405, P<0.01). The area under ROC curve (AUC) and 95%CI predicted by plasma microRNA-214 and BNP were 0.824 (0.757-1.015) and 0.785 (0.721-0.864) in patients with AMI and LVR, which were higher than CRP [0.716 (0.645-0.837), P=0.0167]. The sensitivity and specificity of plasma microRNA-214 in predicting the occurrence of LVR were 72.6% and 86.2% in patients with AMI, respectively. Logistic regression model analysis showed that plasma microRNA-214 and BNP were risk factors of LVR. Conclusion High expression of plasma microRNA-214 is found in LVR patients, which is expected to be the biological indicator for predicting the occurrence of LVR after PCI in patients with AMI.
5.Expressions of Let-7 in Normal Breast Tissue and Breast Cancer Tissue
Zhen YU ; Mingyao XIAO ; Ming CUI ; Min LIU ; Shuying DAI ; Ying CHEN ; Yuefeng HE
Journal of Kunming Medical University 2016;37(7):19-21
Objective To study the expression characteristics of Let-7 genes in breast cancer.Methods Twenty-eight patients with breast cancer were randomly selected,and their cancer tissue and adjacent normal tissue were collected.TRIzol was used to extract the total RNA and real-time quantitative PCR was used to detect the relative gene expression levels.Results The expression of precursor Let-7 in cancer tissue was (9.65 ± 2.31),which was lower than that in normal tissue (10.05 ± 2.81),P =0.048.Precursor Let-7 had dependence relationship with the long menstruation (b =0.816,P =0.029).The menarche age showed a positive correlation with precursor Let-7 in normal tissue (r =0.502,P =0.048) and a negative correlation in cancer tissue (r =-0.484,P =0.049) Conclusions The expression of precursor Let-7 in cancer tissue is lower than that in normal tissue.The period of menstruation is a protective factor to breast cancer.
6.Efficacy of tolvaptan for delusional hyponatremia in decompensated liver cirrhosis
Xiao KAN ; Lei CUI ; Jiachao PAN ; Rong WANG ; Zhaomin ZHENG ; Haitao ZHANG ; Min PENG
Chinese Journal of Infectious Diseases 2013;31(11):658-662
Objective To investigate the efficacy and safety of vasopressin receptor antagonist tolvaptan for treating dilutional hyponatremia casused by decompensated liver cirrhosis.Methods Ninety-six subjects with decompensated liver cirrhosis complicated by dilutional hyponatremia were divided into test group (n =56) and control group (n =40) by double blind method.Test group were treated with a single dose of tolvaptan 15 mg orally in addition to routine therapy,while control group were treated with routine therapy plus placebo.The changes in serum sodium concentration,liver functions,ascites,and edema of lower extremities between the two groups were observed.Measurement data were compared by t test,and categorical data were compared by chisquare test.Results On day 7,36 (64.3%) patients in test group and 9 (22.5%) patients in control group reached normal serum sodium concentrations (x2 =5.241,P<0.01).All the patients in test group and only 3 patients in control group had 24 hours' total urine volume above 3500 mL.Difference of 24 hours' total urine volume during 7-day treatment between the two groups was statistically significant (t=20.899,P<0.01).Thirty-nine patients in test group and 15 patients in control group had reduced ascites (x2 =4.260,P=0.039),but improvement of edema in lower extremities of both groups was comparable (12 cases in test group and 7 cases in control group; x2 =0.227,P=0.634).Serum alanine aminotransferase (ALT),total bilirubin (TBil),potassium and creatinine levels of test group were improved after treatment,but were not significantly different from either baseline levels (t=1.509,0.783,1.107,1.237; both P>0.05) or those of the control group (t=1.712,1.635,1.121,0.873; both P>0.05).Conclusions Single dose of tolvaptan (15 mg) exerts a strong diuretic effect and is able to greatly increase serum sodium concentration,thus has distinct therapeutic effect for patients with decompensated liver cirrhosis complicated by dilutional hyponatremia.
7.Analysis of electroacupuncturing Neiguan points on mitochondrial Ca2+ release and cardiomyocyte apoptosis of diabetic cardiomyopathy in rats
Fengying XIAO ; Jintao CUI ; Yaxi ZHANG ; Jing HE ; Min LI ; Jun ZHANG
International Journal of Traditional Chinese Medicine 2014;(6):544-546
Objective To investigate the influence of electroacupuncturing Neiguan points on mitochondrial Ca2+release and cardiomyocyte apoptosis of diabetic cardiomyopathy in rats. Methods 30 Wistar rats were randomly divided into a control group, a model group and a treatment group(10 in each group). In the treatment group, after eight weeks electroacupuncture Neiguan point treating diabetic cardiomyopathy in rats, the myocardium was quickly removed,mitochondrial was extracted,dual-beam UV spectrophotometer was adopted to detecting Ca2+transportation by testing Ca2+indicator A Ⅲ absorbance, and Cardiomyocyte apoptosis was evaluated by terminal-deoxynucleotidyl transferase mediated deoxy-UTP nick end labeling (TUNEL). Results Ca2+ indicator A Ⅲ absorbance detection(0.051±0.014) and cardiomyocyte apoptosis(0.49±0.36)were more depressed in the treatment group than in the model group[(0.077±0.025), (0.53±0.04)], there were significantly different(P<0.05) Conclusion Effect of Electroacupuncturing Neiguan points decreased myocardial apoptosis index in rats with diabetic cardiomyopathy, which may be related to the reduction of mitochondrial Ca2+influx.
8.Clinical observation of the carotid of patients with anterior ischemic optic neuropathy using doppler ultrasound
Ya-Li, ZHANG ; Xiao-Li, CUI ; Xian-Min, JI ; Ai-Yi, ZHOU
International Eye Science 2015;(7):1269-1270
AlM: To investigate the relationship between the anterior ischemic optic neuropathy ( AlON ) and the carotid artery change using doppler ultrasound.METHODS:Fifty-four cases of AlON patients and 54 cases of healthy control were observed, atherosclerotic spots were detected by the application of color ultrasound.RESULTS:ln AlON group of 54 patients, 38 cases appeared carotid atherosclerosis, accounting for 70%. The number of cases with hard plaque, soft plaque and mixed plaques were 18, 13, and 7 respectively, accounting for 33%, 24% and 13%. ln the control group, 20 cases were detected atherosclerotic change, accounting for 37%. And the number of cases with hard plaque, soft plaque and mixed plaques were 12, 5 and 3 respectively, accounting for 22%, 9%, 6%. Significant stenosis and velocity change were showed in neither AlON group nor control group. Compared with the control group, AlON group had more cases of atherosclerotic plaque, the difference was statistically significant (χ2=12. 836, P=0. 005)CONCLUSlON: The incidence of AlON is correlated with carotid atherosclerosis, and carotid ultrasonography is significantly valuable for AlON etiology and diagnosis.
9.Effects of inward rectifier potassium channel blockers on EPCs function.
Wen-ping LI ; Xiao-dong CUI ; Ning-ning HOU ; Xiao-yun ZHANG ; Jian-hua LIU ; Jing ZHANG ; Min CHENG
Chinese Journal of Applied Physiology 2015;31(5):448-451
OBJECTIVETo investigate the effects of inward rectifier potassium channel blockers (BaCl2, CsCl) on the functions of endothelial progenitor cells (EPCs).
METHODSDensity gradient centrifugation-isolated rat hone marrow mononuclear cells were cultured in vitro. EPCs were harvested and seeded on six culture dish when cells grew to 3-5 passages. Before testing the EPCs were synchronized with M199, which contain 2% fetal calf serum. In the end, EPCs were treated with different intervention. The experiment mainly included two parts: (1) BaCl2 (100 micromol/L) and free BaC2 of Tyrodes solution; (2) CsCl (1 mmol/L) and control. Cell pretreated with blockers above mentioned for 12 h, then the gene expression of stromal cell-derived factor-1 (SDF-1), epoprotenol (PGI2) were assessed, beyond that the ability of adhesion, migration were assayed with different tests. In addition, the medium was collected when EPCs were treated for 3 days. The levels of SDF-1 were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Going even further, EPCs were treated with the signal pathway blockers in advance, after repeat the above steps, in order to analyze the change of SDF-1 and then discuss its mechanism.
RESULTSCompared with control group, BaCl2, CsCl could increase EPC adhesion and migration to same extent. Moreover, the gene expression of SDF-1, PGI2 was significantly up-regulated and the production of SDF-1 increased evidently. Furthermore, the mechanism of SDF-1 secretion increasing mainly was associated with eNOS signaling pathways.
CONCLUSIONBa2+ and Cs+ play important roles in increasing EPCs functions, such as adhesion, migration and secretion.
Animals ; Barium Compounds ; pharmacology ; Cells, Cultured ; Cesium ; pharmacology ; Chemokine CXCL12 ; metabolism ; Chlorides ; pharmacology ; Endothelial Cells ; cytology ; Enzyme-Linked Immunosorbent Assay ; Potassium Channels, Inwardly Rectifying ; antagonists & inhibitors ; physiology ; Rats ; Stem Cells ; cytology
10.Breeding on eight strains of Pseudostellaria heterophylla based on phenotypic traits and quality in Guizhou province.
Hou-Xi XIONG ; Tao ZHOU ; Wei-Ke JIANG ; Min CHEN ; Cui-Cui HUAN ; Chuan-zhi KANG ; Chang-gui YANG ; Cheng-Hong XIAO ; Ming-Wu LIAO
China Journal of Chinese Materia Medica 2014;39(21):4197-4204
OBJECTIVETo provide new germplasm materials for breeding new varieties of Pseudostellaria heterophylla.
METHODThe method of single plant selection was adopted, with the comparative experiments being carried out under the same conditions in Shibing county. The 8 plants of Shibing SB-4 were compared respectively with factor analysis for 27 phenotypic traits and 8 yield traits, and single factor variance analysis for the contents of polysaccharides.
RESULTUsing factor analysis, 27 phenotypic traits were classified into 7 principal divisors and 8 yield traits were simplified into 3 principal divisors. The 4 strains of P. heterophylla, ZT-01, ZT-02, ZT-06 and ZT-07, performed better than others in the phenotypic traits, and ZT-01, ZT-02, ZT-03 and ZT-07 in the yield traits. The contents of polysaccharides of ZT-01, ZT-02, ZT-05 and ZT-08 showed significantly higher value.
CONCLUSIONThere is significant difference among the 8 strains of P. heterophylla in phenotypic traits, yield traits and quality traits, making it possible to select certain strains for different purposes. ZT-01 and ZT-02 can be breaded further. ZT-06 and ZT-07 were used as ornamental cultivars for its great phenotypic traits. ZT-03 with good resistance and high yield was taken as resistant variety, and ZT-05 would face next selection on the basis of its high content of polysaccharide.
Breeding ; Caryophyllaceae ; chemistry ; growth & development ; China ; Phenotype ; Polysaccharides ; analysis