2.Investigation of key miRNAs and their target genes in inflammatory bowel diseases and colitis-associated colorectal cancers using miRNA proifling and bioinformatic tools
Yuan YIN ; Cheng WANG ; Xin DAI ; Zhaohui HUANG
China Oncology 2016;26(11):916-921
Background and purpose:Inlfammatory bowel diseases (IBD) are a group of chronic intestinal diseases, including ulcerative colitis (UC) and Crohn’s disease (CD). This study identified differentially expressed miRNAs in UC, CD and colitis-associated colorectal cancers (CAC) to explore their potential as novel molecular biomarkers. Methods:Tissue samples were taken from 13 UC patients, 3 CD patients, 12 CAC patients, and 8 age-and gender-matched healthy controls. The miRNA expressions were detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) assay. Known targets of deregulated miRNAs were utilized using miRWalk 2.0 database, and subsequent bioinformatics analysis of these target genes was performed by DAVID software (GO-analysis, KEGG-analysis and BIOCARTA-analysis). Results:The data showed that miR-146a, miR-27a, miR-29a, miR-20a and miR-21 were upregulated in UC, CD and CAC tissues compared with normal control. Moreover, the target genes of these miRNAs were enriched in several key signal transduction pathways including cancer-related pathway and immu-nity-associated pathway. Conclusion:miR-146a, miR-27a, miR-29a, miR-20a and miR-21 may play important roles in the switching from IBD to CAC.
3.The effect of pulsed electromagnetic fields on the proliferation of immunomagnetically separated human pre-cartilaginous stem cells
Junfang WANG ; Renyun XIA ; Huang FANG ; Anmin CHEN ; Delong YIN ; Hao CHENG
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(5):296-300
Objective To study the biological effects of pulsed electromagnetic fields (PEMFs) on the pro-liferation of immunomagnetically separated human precartilaginous stem cells (PSCs) in vitro. Methods The cells from an aborted fetus's metaphysis were digested using collagenase. The PSCs were isolated by magnetic cell sorting (MACS), then subcultured and amplified. Flow cytometry, immunohistochemistry, immunofluorescence and RT-PCR analysis were performed to identify the purified PSCs. The PSCs were stimulated by PEMFs at 50 Hz frequency and 1 mT intensity. Cell proliferation was measured at different time points using methyl thiazolyl tetrazolium ( MTT), and the cell growth curve was plotted. Flow cytometry was applied to measure the cell cycle and apoptosis. Results The PSCs were successfully cultured. There was fibroblast growth factor receptor-3 (FGFR-3) on their sur-face. Cell proliferation was promoted after 4 and 6 days of PEMF stimulation. The percentage of cells at the S phase was higher than in a control group. Early, late and total rates of apoptosis in the experimental group decreased signifi-cantly. Conclusion PEMFs can enhance the proliferation and inhibit the apoptosis of human PSCs, and it is possi-ble to cultivate the high density human PSCs in vitro.
4.Comparison of biological characteristics of mesenchymal stem cells derived from bone marrow, peripheral blood and cord blood
Youzhang HUANG ; Jianliang SHEN ; Lizhong GONG ; Wenjie YIN ; Yi LIU ; Hai CHENG ; Peihao ZHENG ; Jian CEN
Chinese Journal of Tissue Engineering Research 2009;13(45):8966-8970
BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P < 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.
5.Inspection application value of digital tomosynthesis in children atlanto-axial joint subluxation
Lichun ZHU ; Chuangao YIN ; Zifeng SHI ; Lei FAN ; Shanfeng HUANG ; Cheng ZHANG
Journal of Practical Radiology 2016;32(4):588-590
Objective To explore the digital tomosynthesis(DTS)technology application value in children with atlanto-axial joint subluxation inspection.Methods 30 children with torticollis after injury,head and neck activities obstacles or local pain were selected.The consent of patient and family was obtained.At the same time the atlanto-axial joint open mouth,cervical lateral radiographies and DTS scanning were performed,two imaging quality and the diagnosis were compared.Results In all 30 cases,excellent imaging rate was 90% (27/30)on DTS and 53% (1 6/30)on DR respectively.On DTS atlanto-axial sublution were found in 22 cases but on DR atlanto-axial sublution were diagnosed in only 12 cases,suspicious subluxation in 4 cases,and misdiagnosis in 6 cases.Conclusion For diagnosis of atlanto-axial joint subluxation in children,DTS can obtain high quality images,clearly display anatomic structures and pathological morphology of atlanto-axial joint.
6.Correlation study among social avoidance and acceptance of disability in burn patients
Zheyuan XIA ; Shuang YU ; Yue KONG ; Tingting YIN ; Suhua SHI ; Rong HUANG ; Yuhong CHENG
Chinese Journal of Practical Nursing 2014;30(31):27-30
Objeetive To investigate the relationship among social avoidance and acceptance of disability in burn patients.Methods Totally 127 patients were investigated using Social Avoidance and Distress Scale (SADS),Acceptance of Disability Scale (AODS),demographic information questionnaire and disease related information sheet.Results The points of burn patients' SADS was (21.38±6.75),One-way ANOVA showed that gender,marital status,special parts burns,amputation affected social avoidance significantly.The total score and factors scores of acceptance of disability were all negatively related to the score of social avoidance.Regression analysis showed that acceptance of disability,head burns,gender were the independent risk factors of depression.Conclusions Acceptance of disability were negatively related to social avoidance; Nurse can help them accept the reality of disability,develop the confidence to partici-pate in social communication,and improve the social adaptability.
7.Characteristics of Neuro-Behcet disease: six cases case report and literature review
Yuhui CHEN ; Yongjing CHENG ; Cibo HUANG ; Yin WANG ; Lianzhe WU ; Tao GONG
Chinese Journal of General Practitioners 2015;14(9):694-697
Six patients with Neuro-Behcet disease (NBD) were admitted in Beijing Hospital from 2000 to 2014,during the same period 344 NBD cases were reported in the literature.The clinical data of 350 Chinese NBD patients,including the symptoms,signs,laboratory tests,imaging,treatment and prognosis were retrospectively analyzed.The average onset age was 36.7 years old.The average interval time from Behcet disease (BD) to NBD was 5.6 years.NBD often occurred in male,the male/female ratio was 1.94:1.The most common manifestations were oral ulcer 94.9% (277/292),followed by genital ulcer 69.1% (215/311) and fever 42.1% (131/311).The most common symptoms in neurology were limb weakness (36.9%,129/350),headache (34.0%,119/350) and sensory disturbance (21.1%,74/350),followed by bulbar symptoms (11.4%,40/350),pyramidal sign was most often (27.1%,95/350).The central neurology system (CNS) was the major target of NBD,parenchymal CNS was affected most (88.0%,308/350),non-parenchymal CNS was affected less (5.7%,20/350),peripheral neurology system (PNS) was rarely affected (6.9%,24/350).Steroids were the first-line drug for NBD,if it failed,immunosuppressive agents or biologic agents could be used.The most NBD patients had single episodes,32.0% (49/153) had relapses with remission,7.7% (27/350) had adverse prognosis.Analysis showed that NBD should be paid more attention.
8.The establishment and application of graded management under early warning for intrahospital transport
Lin WAN ; Suhua SHI ; Rong HUANG ; Yue KONG ; Tingting YIN ; Yuhong CHENG
Chinese Journal of Nursing 2017;52(1):59-62
Objective Through conducting the project of quality improvement for intrahospital transport of critically ill patients from ward to ICU,to establish graded management under early warning to improve transport efficiency and quality.Methods Through setting up project team,setting goals of quality improvement,measuring and analyzing transport status,the graded management under early warning was established from three aspects:condition,equipment and transport personnel.The graded management under early warning was applied to clinical nursing practice to evaluate the effects.Results There was no statistical difference before and after the implementation of graded management under early warning in gender,age and condition of critical ill patients between two groups,but the time of transport was significantly reduced after the implementation,while there was no equipment failure,and the incidence of adverse events associated with devices decreased effectively.Conclusion The establishment and application of graded management under early warning has effectively reduced the risks of transport,improved efficiency and quality of transport.
9.Prediction and analysis of the subcellular localization of Arnt2 in rat cerebellar granule neurons
Linguang SUN ; Wei YIN ; Yijun HUANG ; Wenfang CHENG ; Xingwen SU ; Pengxin QIU ; Guangmei YAN
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.
10.Study of the expression of the cyclin D1 in chronic myelogenous leukemia
Liefen YIN ; Ling YANG ; Mengmeng CUI ; Zhenglei SHEN ; Xiaoyuan QU ; Lifang CHENG ; Guiyun HUANG
Journal of Leukemia & Lymphoma 2008;17(3):187-188,191
Objective To examine the expression levels of cyclin Dl in the patients with chronic myelogenous leukemia (CML), and evaluate the pathogenesis and clinical significance of cyclin Dl in CML Methods The real-time quantitative polymerase chain reaction (RQ-PCR) was performed to detect the expression levels of cyclin Dl in the bone marrow samples of 18 patients with CML, and 16 samples of benign hemopoietic patients. The relationship between the expression levels of cyclin Dl and the progression and prognosis of patients with CML were analyzed. Results The level of cyclin Dl was higher expressed in 18 patients with CML than the control group (P <0.001). The levels of cyclin Dl was apparently higher expressed in accelerated phase /blast crisis phase than in chronic phase (P <0.05). And the RQ-PCR method showed the tendency that a significant increase was observed in the levels of cyclin Dl from 0.1980 in control group to 1.4002 in chronic phase and 5.4540 in accelerated phase /blast crisis phase. Conclusion The cyclin Dl overexpressed in CML, the roles of cyclin Dl in CML might be an oncogene expressed. The expression level is correlated with the progression and prognosis of patients with CML.