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.Physicochemical properties of medicinal fungus Polyporus umbellatus sclerotial exudate.
Yong-Mei XING ; Hong-Lian LI ; Shun-Xing GUO
China Journal of Chinese Materia Medica 2014;39(1):40-43
This study was conducted to investigate the physicochemical properties of Polyporus umbellatus sclerotial exudate. Morphological characteristics of the sclerotia and its exudate were observed during different stages of sclerotial formation. The pH of the exudate was detected at different time during cultivation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of P. umbellatus sclerotial exudate during cultivating time. Additionally, the protein content was measured by means of BCA protein assay. Furthermore, CAT content was detected using ultraviolet absorption method. That the protein content of the exudate and CAT specific activity rose gradually during the passage of the cultivating time indicated a high level of oxidative stress during P. umbellatus sclerotial exudate formation. The results showed that the pH of the exudate increased gradually and then dropped down during sclerotial formation. That the pH of the exudate maintained the acidity state during the cultivation indirectly indicated that acidic environment would help sclerotial formation. The exudate produced gradually and was absorbed by the sclerotia itself.
Culture Media
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chemistry
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metabolism
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Fungal Proteins
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chemistry
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metabolism
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Fungi
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chemistry
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metabolism
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Hydrogen-Ion Concentration
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Medicine, Chinese Traditional
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methods
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Oxidative Stress
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Polyporus
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chemistry
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metabolism
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Polysaccharides
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chemistry
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metabolism
3.Prokaryotic soluble expression, purification and function study of LEDGF/p75 protein.
Da-Wei ZHANG ; Hong-Qiu HE ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2014;49(8):1200-1207
HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular
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Escherichia coli
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metabolism
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HIV Integrase
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metabolism
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HIV-1
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physiology
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Humans
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Intercellular Signaling Peptides and Proteins
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biosynthesis
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Protein Binding
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Virus Replication
4.Studies on twelve cytokines in serum of coronary heart disease patients
Xing-Wang JIA ; Ya-Ping TIAN ; Guang-Hong GUO ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective The significance of serum cytokine profile in coronary heart disease patients have been evaluated by observing the variation cytokines Methods The serum IL-1 a,IL-1 ?,IL-2,IL-4, IL-6 ,IL-8,IL-10,VEGF,IFN-?,,TNF-?,MCP-1 and EGF in 76 patients and 26 matched healthy volunteers have been observed.Serum cytokines had been measured by protein chips methods in EVIDENCE 180 automatic biochips analyzer and cytokine profile were got by bioinformatics.Results The serum level of IL-1?,IL-6,IL-8,and TNF-? were (4.32?8.14) ng/L ,(13.16?16.81) rig/L,(375.53?292.14) ng/L,(15.46?15.38) ng/L respectively,which increased obviously in patients (P
5.Related complications of implantable venous access port
Hong LIU ; Guo SHI ; Lei XING ; Feng LUO
Journal of Regional Anatomy and Operative Surgery 2017;26(9):648-651
Objective To explore the incidence and clinical treatment of related complications caused by implantable venous access port(IVAP) in patients with breast cancer during chemotherapy.Methods The data of 755 patients with breast cancer recieved chemotherapy by which caused some related complications in our hospital from January 2014 to March 2016 were retrospectively analyzed.Results 753 patients IVAPs were implanted succussfully.The total placement time of implantable venous access port was from 110 days to 940 days,with median placement 147.33 days.The related complications of IVAP were catheter malposition(0.79%,6/755),catheter-related thrombosis(27.81%,210/755),catheter fracture(0.13%,1/755),port exposure(0.93%,7/755) and IVAP-related bloodstream infection(0.13%,1/755).The IVAP-related complications and thrombosis rate were significant higher when IVAPs implanted in the left internal jugular veincompared with that in right internal jugular vein(34.88% vs.25.74%,33.10% vs.24.68%).Conclusion Application of IVAP in patients with breast cancer during chemotherapy is a safe and effective operation.The most common complication is asymptomatic mural thrombus formation around the catheter,which should be paid attention to.
6.Assessment of overall function after percutaneous coronary intervention by cardiopulmonary exercise testing in patients with stable coronary heart disease.
Hong-chao ZHENG ; Yue-you DING ; Xing-guo SUN ; Jian YANG ; Qing LI ; Fang LIU
Chinese Journal of Applied Physiology 2015;31(4):378-382
UNLABELLEDObjective: In order to assess the integrative cardiopulmonary function after percutaneous coronary intervention (PCI) in patients with stable coronary artery disease (CAD), we used symptom limited maximum cardiopulmonary exercise testing (CPET).
METHODSAll 59 patients diagnosed stable CAD by coronary angiography and echocardiography from August to December of 2014 in our hospital, were divided two groups. PCI group, 31 patients received PCI and drugs. Control group, 28 patients received drugs therapy only. All patients performed CPET before and after the treatment.
RESULTSAll patients safely completed CPET without any complications. The control group, all functional parameters were unchanged (P > 0.05). PCI group, the anaerobic threshold, peak oxygen uptake and peak oxygen pulse increased significantly (P < 0.05) from baseline,but not for others (P > 0.05). For individual analysis, PCI group had higher rates of increase (≥ 10% of baseline) in both peak oxygen uptake and peak oxygen pulse than those of control group (P < 0.05).
CONCLUSIONCPET is an objective, quantitative, safe and effective method to evaluate the clinical therapeutic efficiency. PCI can improve the integrative cardiopulmonary function in CAD patients.
Anaerobic Threshold ; Coronary Angiography ; Coronary Artery Disease ; surgery ; Exercise Test ; Heart Rate ; Humans ; Oxygen ; Oxygen Consumption ; Percutaneous Coronary Intervention
7.Study on mechanism of combined administration of Coptidis Rhizoma and Rehmanniae Radix in treating type II diabetes mellitus.
Jing WANG ; Zi-Min YUAN ; Yun-Xing LI ; Hong-Wei KONG ; Guo-Wang XU
China Journal of Chinese Materia Medica 2014;39(3):526-530
To make a preliminary study on the mechanism of Coptidis Rhizoma(CR) and Rehmanniae Radix(RR) before and after the combined administration in treating type II diabetes mellitus. The type I diabetes animal model in rats was established by fat emulsion and intraperitoneal injection with streptozotocin, in order to compare the hpyerglycemic and hypolipidemic effects of CR, RR and their combined administration of different ratio. The urinary metabolic profiling in rats of Coptidis Rhizoma and Rehmanniae Radix before and after the combined administration was analyzed by using the gas chromatography-mass spectrometry. The differences among groups in metabolome were analyzed by the principal component analysis (PCA). The biochemical index results indicated that both CR and RR before and after the combined administration could lower high blood glucose, hypertriglyceride and high cholesterol. According to the analytical results of PCA of the rats' urine samples, the CR group was the most close to the normal group, with no significant difference in CR and RR group of different combination ratios. Twelve differentiated metabolites were identified to be related to type II diabetes. Compared with the normal group, the CR-treated group showed significant increase in seven differentiated metabolites. Among CR and RR drugs with different combination ratios, CR played a major role and thus acted as the monarch drug. Whereas RR served as the ministerial drug and assisted CR to show the efficacy. This study laid a foundation for the explanation of the combination mechanism of traditional Chinese medicines.
Animals
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Blood Glucose
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metabolism
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Diabetes Mellitus, Type 2
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blood
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drug therapy
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urine
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Drugs, Chinese Herbal
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pharmacology
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therapeutic use
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Male
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Rats
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Rats, Wistar
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Rehmannia
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chemistry
8.Expression profiles analysis of two member of squaleneepoxidase gene family from Eleutherococcus senticosus.
Yue-Hong LONG ; Fei-Fei LI ; Guo YANG ; Zhao-Bin XING
China Journal of Chinese Materia Medica 2015;40(1):59-62
In order to find the characteristics of two members of gene family of squaleneexpoxidase (SE) , a quantitative real time PCR method was developed to analyze the expression of Eleutherococcus senticosus SE1 and SE2 gene from different growth periods and in different organs. The result indicated that all the expression of SE2 more than SE1 in the whole growth period and organs of E. senticosus. And in the whole growth period, expression of SE1 showed a low-high-low characteristic. Both expression of SE2 and growth period showed the same trend. The lowest content of the expression was in the roots. SE1 expression have been improved more than SE2 when treated with MeJA. The expression of E. senticosus SE1 and saponins content had significantly positive correlation (P < 0.05) and the correlation coefficients was 0. 858, while the correlation was not significant for SE2. That indicated that SE1 played a key enzyme gene in the biosynthesis of triterpenoidsaponins
Eleutherococcus
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chemistry
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enzymology
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genetics
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growth & development
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Gene Expression Regulation, Plant
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Peroxidase
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genetics
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metabolism
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Plant Proteins
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genetics
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metabolism
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Saponins
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analysis
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metabolism
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Transcriptome
9.Expression and clinical significance of lymphoid differentiation antigens in acute myeloid leukemia.
Journal of Experimental Hematology 2004;12(3):329-331
The present study was aimed to elucidate the expression features of lymphoid differentiation antigens and their clinical implications in acute myeloid leukemia (AML). Immunophenotypes were examined by indirect immunofluorescence method with monoclonal antibodies (McAb) in 62 patients with AML. The results showed that 11 cases of AML were found to express lymphoid differentiation antigen Ly(+)AML in addition to myeloid antigen expression. There was no significant difference in clinical manifestation and blood test between the groups of Ly(+)AML and Ly(-)AML when they were newly diagnosed. Only one case gained long-term remission using standard chemotherapy. It is concluded that Ly(+)AML cells seem to be not sensitive to conventional chemotherapy, however, a treatment protocol for both ALL and AML can improve the curative effects. The overexpression of CD34(+) may be responsible for the relatively low level of long term survival rate in Ly(+)AML patients.
Adolescent
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Adult
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Aged
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Antigens, Differentiation
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analysis
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Antigens, Ly
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analysis
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Female
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Fluorescent Antibody Technique, Indirect
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Humans
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Immunophenotyping
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Leukemia, Myeloid, Acute
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drug therapy
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immunology
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pathology
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Male
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Middle Aged
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Prognosis
10.Time tendency and its cohort study of leukemia mortality of residents in Urumqi.
Journal of Experimental Hematology 2004;12(5):694-696
To explore the distribution of leukemia mortality and its relationship with age of residents who live in city Urumqi, data of leukemia mortality of Urumqi residents from 1985 years to 2000 years was analysed. On the basis of multiple cross-sectional analysis,a cohort study was applied to further analysis. The results showed that the total mortality and male's mortality of leukemia in Urumqi residents were almost the same as that of national level, while the female's mortality of leukemia was lower than that of national level. The cohort study showed that mortality of adolescent with leukemia descended with the increase of age, but mortality of gerontic leukemia ascended with the increase of age. In conclusion, there is a significant relationship between the morbific age of leukemia and prognosis of patients with leukemia.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Child
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Child, Preschool
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China
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epidemiology
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Cohort Studies
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Female
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Humans
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Infant
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Infant, Newborn
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Leukemia
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mortality
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Male
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Middle Aged
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Time Factors