1.Long-term in vitro culture and hepatocytic differentiation of human umbilical cord-derived mesenchymal stem cells
Zhengyan ZHU ; Junqing YAN ; Tao HAN ; Zhi DU ; Ying LUO ; Peng WANG ; Yingtang GAO ; Tong LIU
Chinese Journal of Tissue Engineering Research 2009;13(49):9792-9796
OBJECTIVE: To investigate biological characteristics of human umbilical cord-derived mesenchymal stem cells, and to explore the possibility of hepatocyte-like cells differentiation.METHODS: The umbilical cord was provided by healthy term birth woman in Tianjin Third Central Hospital. Mesenchymal stem cells were isolated from human umbilical cord by enzyme digestion method. Cells were passaged at 80%-90% confluent. The ninth passage of cells at a density of 5×10~(10)/L were seeded in 12-well culture plate and incubated with DMEM containing hepatocyte growth factor, fibroblast growth factor-4 and oncostatin for 28 days. Cell growth activity was detected by MTT method; cell cycle was detected by flow cytometry; surface immunological marker in MSC was detected by immunocytochemical stain and flow cytometry; specific surface phenotype of hepatocyte was detected by immunocytochemical staining. Function characteristic of hepatocyte was determined by staining for glycogen.RESULTS: MSCs were isolated from human umbilical cord and presented with fibroblastic morphology. 80% of cells were at G_0/G_1 phase with good growth activity and stably passaged over 20 times. These cells were positive for CD29, CD105, and Vimentin, but negative for CD34 and CD31. MSCs were induced to hepatocyte-1 ike cells that were positive for alpha fetoprotein, CK18, CK19 at 1 week and albumin at 3 weeks. At 4 weeks, induced cells were positive for glycogen staining.CONCLUSION: MSCs isolated from human umbilical cord can be cultured in a long periods time in vitro and are able to differentiate into functional hepatocyte-like cells.
2.Establishment of the limited cell line of skin fibroblast and their differentiation to hepatic cells
Zhengyan ZHU ; Zhi DU ; Ying LUO ; Jiao LI ; Linlin SUN ; Peng WANG ; Tong LIU ; Yingtang GAO
International Journal of Biomedical Engineering 2011;34(5):284-287,292
Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3~30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5%.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.
3.A new caffeate compound from Nardostachys chinensis.
Ying-peng CHEN ; Zhong-ping WANG ; Hong-hong ZHENG ; Yan-tong XU ; Yani ZHU ; Peng ZHANG ; Hong-hua WU
Acta Pharmaceutica Sinica 2016;51(1):100-104
A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including (+)-licarin A (2), naringenin 4', 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this is the first report of compounds 2, 5 and 6 from Nardostachys genus.
Caffeic Acids
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chemistry
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isolation & purification
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Flavanones
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chemistry
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isolation & purification
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Furans
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chemistry
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isolation & purification
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Glucosides
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chemistry
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isolation & purification
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Hydroxybenzoates
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chemistry
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isolation & purification
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Lignans
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chemistry
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isolation & purification
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Nardostachys
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chemistry
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Plant Roots
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chemistry
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Rhizome
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chemistry
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Vanillic Acid
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chemistry
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isolation & purification
4.Multiplex sequence-specific PCR detection for simultaneous screening of 5 types of JAK2 mutations in myeloproliferative diseases
Hongxing LIU ; Chunrong TONG ; Peng CAI ; Guirong TANG ; Xian ZHANG ; Wen TENG ; He WANG ; Ying ZHANG ; Ping ZHU
Chinese Journal of Laboratory Medicine 2009;32(1):30-34
Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.
5.Content Determination of Polyphyllin Ⅰ and Ⅱ in Kangkeling Mixture by HPLC
Min PENG ; qin Mei SHI ; dong Xiao TONG ; ying Shui YANG ; Yan XU
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(11):75-77
Objective To establish a method for determining the contents of polyphyllin Ⅰ and polyphyllin Ⅱ in Kangkeling Mixture. Methods The contents of polyphyllin Ⅰ and Ⅱ were determined by HPLC gradient elution. Poroshell 120 Ec-C18 column (4.6 mm × 150 mm, 4 μm) was used; Acetonitrile-water (A:B) was set as the mobile phase; the flow rate was 1.0 mL/min; the detection wavelength was 210 nm; column temperature was 30 ℃. Results Polyphyllin Ⅰ showed good linear relationship in the range of 1.009–10.09 μg (r=0.999 6), and the average recovery was 97.31% (RSD=2.05%, n=6). Polyphyllin Ⅱ showed good linear relationship in the range of 0.640 5–6.405 μg (r=0.999 8), and the average recovery was 96.41% (RSD=1.67%, n=6). Conclusion The method is simple, with good repeatability and accurate results, which can be used to determine the contents of polyphyllin Ⅰ and Ⅱ in Kangkeling Mixture.
6.Analysis on the rodents density monitoring data in Fangshan District of Beijing, 2016-2017
Hai-ying GONG ; Zhao-hui LIU ; Ying TONG ; Yong ZHANG ; Mei-de LIU ; Xiao-peng ZENG
Shanghai Journal of Preventive Medicine 2020;32(12):992-
Objective To understand species, population density, and seasonality of rodents in Fangshan District of Beijing and provide the information for prevention and control of rodents and rodent-borne diseases. Methods The rodents were captured by using the trap at-night method in Mukezhai, Xianfenggu, Liushilu, Gushanzhai, Wanjingxiangou.The rodents pieces were identified and the density was calculated. Results Totally 4 665 traps were placed and 268 rodents were captured during 2016-2017.Among the seven identified species, the proportion of
7.Impact of Qi-invigorating traditional Chinese medicines on intestinal flora: A basis for rational choice of prebiotics.
Xiao-Meng WANG ; Xiao-Bo LI ; Ying PENG
Chinese Journal of Natural Medicines (English Ed.) 2017;15(4):241-254
According to the theory of traditional Chinese medicine (TCM), Qi (vital energy) is regarded as a driving force of biological activities in human body, including both nutrient substances and organ functions. Qi-invigorating TCMs are widely used to treat various symptoms and disorders, such as fatigue, obesity, immunosuppression, intestinal flora imbalance, and gastrointestinal diseases, in which Qi is considered to be reduced or depleted. Interestingly, abundant clinical evidences suggest that these disorders are associated with the alternation of intestinal flora, which directly affects disease status. Herein we review the interaction between gut microbiota and Qi-invigorating TCMs under healthy and disease conditions and discuss the mechanisms of action and applications of Qi-invigorating TCMs in enhancing health status through microbial alternation. A better understanding of the role of Qi-invigorating TCMs in modulating microbial composition and the association between intestinal microbiota and diseases would help reveal the clinical consequences of microbiota alteration and explore opportunities to harness this symbiotic relationship to improve public health.
Drugs, Chinese Herbal
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pharmacology
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Gastrointestinal Microbiome
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drug effects
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Humans
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Prebiotics
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Qi
8.Correlation of JAK2V617F mutation burden with clinical features in patients with polycythemia vera and essential thrombocythemia.
Hong-Xing LIU ; Chun-Rong TONG ; Peng CAI ; Wen TENG ; He WANG ; Ying ZHANG ; Ping ZHU
Journal of Experimental Hematology 2009;17(3):742-745
This study was aimed to analyze the correlation of JAK2V617F mutation burden with clinical features in patients with polycythemia vera (PV) and essential thrombocythemia (ET), The JAK2V617F mutation ratios in 47 PV samples and 43 ET samples were detected by real-time PCR. The correlation of mutation allele ratio in PV and ET samples with clinical features (hemoglobin, hematocrit, white blood cell count and platelet count) was analyzed. The results showed that the JAK2V617F mutation burden was higher in PV (0.441 +/- 0.270) than that in ET (0.209 +/- 0.192). The JAK2V617F mutation burden was positively correlated with levels of hemoglobin (PV: R = 0.518, p < 0.001; ET: R = 0.528, p = 0.005), hematocrit (PV: R = 0.510, p < 0.001; ET: R = 0.524, p = 0.005) and leukocyte (PV: R = 0.584, p = < 0.001; ET: R = 0.471, p = 0.013) in PV and ET samples. The higher JAK2V617F mutation burden was negatively correlated with levels of platelet count in PV samples (R = -0.354, p = 0.020), but there was no correlation between the JAK2V617F mutation burden and platelet count in ET samples (R = 0.233, p = 0.242). It is concluded that the higher JAK2V617F mutation burden is related with higher hemoglobin, hematocrit and leukocyte count in both PV and ET samples. The higher JAK2V617F mutation burden is correlated with lower platelet count in PV samples, but there is no correlation between JAK2V617F mutation burden and platelet count in ET samples.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Case-Control Studies
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Female
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Hemoglobins
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Humans
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Janus Kinase 2
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genetics
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Male
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Middle Aged
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Mutation
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Polycythemia Vera
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genetics
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Polymerase Chain Reaction
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Thrombocythemia, Essential
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genetics
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Young Adult
9.Radiographic predictors of residual low back pain after laminectomy for lumbar canal stenosis: a minimum of 6-year follow-up.
Ying-peng XIA ; Tian-tong XU ; Qing-feng SHEN ; Xue-li ZHANG ; Han JIANG ; Rong TIAN
Chinese Journal of Traumatology 2008;11(3):135-140
OBJECTIVETo identify radiographic predictors of residual low back pain (LBP) after laminectomy for lumbar canal stenosis (LCS).
METHODSClinical results and radiographic findings in 69 patients who underwent single level laminectomy for LCS were retrospectively reviewed. Patients who had an improvement in LBP scores evaluated by Japanese Orthopaedic Association (JOA) scoring system during the follow-up periods were classified as the recovery group, and others were classified as the non-recovery group. Patients'clinical data and radiographic parameters like lordosis angle, range of motion and intervertebral rotational angle were analyzed using binary logistic regression analysis to detect factors significantly related with the occurrence of residual LBP.
RESULTSThe average preoperative JOA score of 14.8+/-5.05 improved to 21.59+/-5.51 at the final follow-up. Binary logistic regression analysis revealed that significant predictors of residual LBP were preoperative lumbar lordosis angle and range of motion.
CONCLUSIONSOur results suggest that patients with flat back and limited lumbar mobility before surgery tend to have poor results in terms of LBP. Therefore, these sagittal radiographic parameters should be taken into account when choosing laminectomy as the surgical option for LCS.
Aged ; Female ; Humans ; Laminectomy ; Low Back Pain ; diagnostic imaging ; Lumbosacral Region ; Male ; Prognosis ; Radiography ; Retrospective Studies ; Spinal Stenosis ; surgery ; Spine ; diagnostic imaging ; Treatment Outcome
10.Real-time three dimensional echocardiography-based evaluation of left ventricular function in children with Kawasaki disease.
Xiao-Xu CHEN ; Ling ZHANG ; Kun WANG ; Ying FU ; Peng-Xiang ZHOU ; Xin-Tong ZHOU
Chinese Journal of Contemporary Pediatrics 2014;16(3):268-271
OBJECTIVETo evaluate the clinical significance of full volume real-time three-dimensional echocardiography (RT-3DE) in the assessment of general and local systolic functions of the left ventricle in children with Kawasaki disease (KD).
METHODSA total of 73 KD children (40 with and 33 without coronary artery lesions) and 35 healthy control children were recruited. Left ventricular ejection fraction (LVEF) was measured by M-mode ultrasound and full volume RT-3DE imaging. A left ventricular volume-time curve and a segmental speed-time curve were generated. Differences between control subjects and patients with and without coronary artery lesions were analyzed.
RESULTSThe M-mode ultrasound measurements of LVEF in KD patients with coronary artery lesions were significantly lower than in KD patients without coronary artery lesions and control children (P<0.05), while there was no significant difference between KD patients without coronary artery lesions and control children. RT-3DE measurements of LVEF were significantly different between the three groups analyzed (P<0.05): coronary artery lesion group < no coronary artery lesion group < control group. RT-3DE-based segmental ventricular wall analysis revealed that Tmsv16-SD and Tmsv12-SD in KD patients with coronary artery lesions were significantly higher than other two groups and Tmsv6-SD was also significantly higher than in the normal control group (P<0.05) and that Tmsv16-SD in KD patients without coronary artery lesions increased significantly compared with the normal control group (P<0.05).
CONCLUSIONSRT-3DE can be used in the quantitative evaluation of the left ventricular function and therefore has significant clinical implications.
Child ; Child, Preschool ; Echocardiography, Three-Dimensional ; methods ; Female ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; diagnostic imaging ; physiopathology ; Ventricular Function, Left