1.Co-culture of adipose-derived stem cells and osteoblasts under different conditions
Yang ZHANG ; Dacheng LIU ; Xiaoning YANG
Chinese Journal of Tissue Engineering Research 2014;(37):6003-6007
BACKGROUND:After co-culture with osteoblasts, bone marrow stem cells can be induced to differentiate into osteoblasts. Whether adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts or not? OBJECTIVE:To observe whether adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts. METHODS:Adipose-derived stem cells and osteoblasts were isolated from New Zealand white rabbits. Then, passage 3 adipose-derived stem cells were co-cultured with passage 2 osteoblasts in 10%or 5%fetal bovine serum for 14 days. RESULTS AND CONCLUSION:After 7 days of co-culture, some adipose-derived stem cells became round in the two groups. After 14 days of co-culture, adipose-derived stem cells highly differentiated and differentiated cells were similar to mature osteoblasts that were positive for alkaline phosphatase staining and alizarin red staining. The mRNA expression of type I col agen and osteocalcin increased in both two group, especial y in the 10%fetal bovine serum group. These findings indicate that adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts induced by high-concentration serum culture.
2.Research progress and trend analysis of biology and chemistry of Taxus medicinal resources.
Dacheng HAO ; Peigen XIAO ; Yong PENG ; Ming LIU ; Li HUO
Acta Pharmaceutica Sinica 2012;47(7):827-35
Taxus is the source plant of anti-cancer drug paclitaxel and its biosynthetic precursor, analogs and derivatives, which has been studying for decades. There are many endemic Taxus species in China, which have been studied in the field of multiple disciplines. Based on the recent studies of the researchers, this review comments on the study of Taxus biology and chemistry. The bibliometric method is used to quantify the global scientific production of Taxus-related research, and identify patterns and tendencies of Taxus-related articles. Gaps are present in knowledge about the genomics, epigenomics, transcriptomics, proteomics, metabolomics and bioinformatics of Taxus and their endophytic fungi. Systems biology and various omics technologies will play an increasingly important role in the coming decades.
3.Calcitonin gene-related peptide induces the osteogenic differentiation of adipose derived stem cells combined with calcium alginate gel
Changzhi HUANG ; Xiaoning YANG ; Dacheng LIU ; Yigong SUN ; Xingming DAI
Chinese Journal of Tissue Engineering Research 2013;(42):7356-7362
BACKGROUND:Calcitonin gene-related peptide has been confirmed to induce osteogenic differentiation, but whether it can induce the osteogenic differentiation of adipose-derived stem cells under three-dimensional culture to construct tissue-engineered bone is rarely reported.
OBJECTIVE:To investigate the feasibility of exogenous calcitonin gene-related peptide to induce osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel in three-dimensional condition.
METHODS:Adipose-derived stem cells were gained by col agenase I digestion of the subcutaneous adipose tissue of both inguinal regions of New Zealand rabbits. Passage 3 cells were mixed with sodium alginate to prepare calcium alginate gel, and then the cells were assigned into two groups and cultured in 24-wel plates. Adipose-derived stem cells in the control group were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 supplemented with 10-2 mol/Lβ-glycerophosphate sodium, 10-7 mol/L dexamethasone, 50 mg/L ascorbic acid, 10%fetal bovine serum. While, adipose-derived stem cells in the experimental group were incubated with the same medium as above, but 1.5 μg/L calcitonin gene-related peptide was added. The cells proliferation and the mRNA expressions of col agen I and osteocalcin were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and reverse transcription-PCR respectively, and alkaline phosphatase and calcium concentration were detected at different induction time.
RESULTS AND CONCLUSION:The cellproliferation curves were S shaped. The absorbance values of the experimental group were higher than those of the control group at1, 3, 5, 7, 14, 21 days after osteogenic induction (P<0.05). Alkaline phosphatase and alizarin red staining of adipose-derived stem cells was al positive, but golden round nodes became more and bigger in the experimental group compared with the control group after 2 weeks. At 7 and 14 days, col agen I and osteocalcin mRNA expressions were higher in the experimental group than in the control group. Alkaline phosphatase activity and calcium concentration of the experimental group were higher than those of the control group at 1, 2, 3, 4 weeks after osteogenic induction (P<0.05). Results showed that the calcitonin gene-related peptide can induce the osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel.
4.Pharmaphylogeny vs. pharmacophylogenomics: molecular phylogeny, evolution and drug discovery.
Dacheng HAO ; Peigen XIAO ; Ming LIU ; Yong PENG ; Chunnian HE
Acta Pharmaceutica Sinica 2014;49(10):1387-94
With the surge of high-throughput sequencing technology, it is becoming popular to perform the phylogenetic study based on genomic data. A bundle of new terms is emerging, such as phylogenomics, pharmacophylogenomics and phylotranscriptomics, which are somewhat overlapping with pharmaphylogeny. Phylogenomics is the crossing of evolutionary biology and genomics, in which genome data are utilized for evolutionary reconstructions. Pharmaphylogeny, advocated by Prof. Pei-gen Xiao since 1980s, focuses on the phylogenetic relationship of medicinal plants and is thus nurtured by molecular phylogeny, chemotaxonomy and bioactivity studies. Phylogenomics can be integrated into the flow chart of drug discovery and development, and extend the field of pharmaphylogeny at the omic level, thus the concept of pharmacophylogenomics could be redefined. This review gives a brief analysis of the association and the distinguished feature of the pharmaphylogeny related terms, in the context of plant-based drug discovery and sustainable utilization of pharmaceutical resource.
5.Characteristics of lymph nodes on 18F-FDG PET/CT imaging in patients with non-Hodgkin's lymphoma and widespread lymph node metastases carcinoma
Qingqing LIU ; Zhenguang WANG ; Nan WANG ; Mingming YU ; Dacheng LI ; Simin LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2016;36(2):142-145
Objective To compare the characteristics of lymph nodes on 18F-FDG imaging in patients with NHL and widespread lymph node metastases carcinoma (WLNMC) for helping the differential diagnosis.Methods A total of 40 NHL patients(22 males,18 females,average age 51 years) and 42 WLNMC patients (19 males,23 females,average age 61 years) confirmed by histopathology from November 2010 to May 2015 were included in this retrospective study.The regions and the metabolism features of the lesions on PET/CT images were observed and recorded with routine visual method.Two-sample t test and x2 test were used for statistical analysis.Results Lymph nodes of NHL were kidney-shaped (47.2%,142/301),which was significantly different from that of WLNMC with circular form (47.6%,140/294;x2 =36.261,P<0.05).Lymph nodes of NHL mostly showed homogeneous density with no necrosis (51.8%,156/301);however,that of WLNMC showed uneven density (20.4%,60/294) or with necrosis (32.3%,95/294;x2 =26.266,P<0.05).Both of the two kinds of lymph nodes showed the characteristic of fusion (30.6% (92/301) vs 36.1% (106/294);x2 =2.019,P>0.05).The ratios of lymph nodes with clear boundary in NHL and WLNMC were significantly different:28.2%(85/301) vs 19.4%(57/294);x2 =6.413,P<0.05.Lymph nodes with symmetric distribution were more in NHL patients (52.5%,158/301)than that in WLNMC patients (42.2%,124/294;X2=6.347,P<0.05).Sizes of lymph nodes in NHL patients and WLNMC patients were not significantly different:(2.08±0.65) cm vs (1.97±0.81) cm;t=0.316,P>0.05.SUVmax of lymph nodes in NHL patients (9.02±3.97) was much higher than that in WLNMC patients (6.92± 1.34;t=0.370,P<0.05).Conclusions Some characteristics of lymph nodes on 18F-FDG PET/CT imaging,such as the distribution,morphology,density and glucose metabolism features,are different in patients with NHL and WLNMC.Those differences may be helpful for the differential diagnosis.
6.Transthyretin-down-regulation in sera of cholangiocarcinoma
Liyun LIU ; Bo LIU ; Xiaomin WANG ; Songwei DAI ; Lingyun HUANG ; Xueyuan XIAO ; Dacheng HE
Chinese Journal of Hepatobiliary Surgery 2011;17(2):127-131
Objective As SELDI-TOF-MS (Surface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) has been broadly used to screen biomarkers for a variety of diseases, the identification and validation of the revealed biomarkers requires more focused attention.Method In this paper, the serum samples from 60 cholangiocarcinoma, 146 lung cancer, 65 LGC and 58 LPC, 49 benign diseases of hepatobiliary and 53 normal individuals were analyzed by SELDI-TOF-MS. Results Among a set of proteins automatically selected as specific biomarkers by Biomarker Wizard software, three protein peaks, with molecular weights of 13. 71 × 103 , 13.83 × 103 and 13. 99 ×103 , were found significantly decreased in cholangiocarcinoma samples. The candidate biomarkers obtained from Tricine-SDS-PAGE gel bands by matching the molecular weight with peaks on CM10 chips were identified by Mass spectrometry as the native transthyretin(native TTR),cysTTR and glutTTR.These preliminary results were further proven by immunoprecipitation using commercial TTR antibodies. This allowed us to re-measure the TTR levels in all the groups more simply by ELISA assay. It showed a firm consistency between ELISA and SELDI analysis. In addition, while TTR levels in cholangiocarcinoma were found to be lower than those in normal healthy controls, TTR levels in benign diseases of the hepatobiliary system were found to be higher than those in healthy controls.Conclusion TTR could be a biomarker that better discriminates cholangiocarcinoma patients from the benign diseases compared to other biomarkers presently available.
7.Effect of simvastatin on bone formation and osteogenic differentiation of bone marrow stromal cells in young rats
Xiaoning LIU ; Liu ZHANG ; Faming TIAN ; Hui ZHANG ; Dacheng HAN ; Junqiang NIU ; Lei ZHANG
Chinese Journal of Tissue Engineering Research 2010;14(7):1152-1156
BACKGROUND: Recently simvastatin has been shown to stimulate osteogenic differentiation and bone formation, but there is no report about the effect of simvastatin on the bone development of young rats.OBJECTIVE: To evaluate the effects of simvastatin on osteogenic relative genes of proximal tibia trabecular bone and osteogenic differentiation of bone marrow stromal cells (BMSCs).METHODS: Twenty 1-week-old Spragua-Dawley young rats were randomly and equally divided into simvastatin and control groups. Rats in the simvastatin group were treated with a subcutaneous injection of simvastatin[5 mg/(kg·d)] for 2 weeks, while rats in the control group were treated with placebo for 2 weeks. The expressions of bone morphogenetic protein-2 (BMP-2), matrix metalloproteinase-13 (MMP-13), and vascular endothelial growth factor (VEGF) of trabecular bone in the tibia were analyzed by mmunohistochemicel staining. BMSCs harvested from the rat femur were osteogenic-differentiation cultured. Alkaline phosphatase (ALP) staining was performed on day 14, real-time PCR analysis was applied to investigate the BMP2, RUNX2,Osterix, MSX2, DLX3, DLX5 mRNA expressions during osteogenic differentiation in vitro on day 21, and von Kossa staining was detected on day 28.RESULTS AND CONCLUSION: ① There was no significant difference in the expressions of BMP-2, MMP-13, and VEGF between simvastatin and control groups. ② The percentages of ALP positive-stained cells were about 30% and there was no significant difference between the two groups (P > 0.05). ③There was no significant difference in the expressions of BMP-2,RUNX2, Osterix, MSX2, DLX3, DLX5 mRNA in osteoganic differentiation-induced BMSCs. ④ von Kossa staining demonstrated that dark brown calcified spots in various sizes were observed, but there was no significant difference in size and density between simvastatin and control groups. A subcutaneous injection of simvastatin[5 mg/(kg·d)] for 2 weeks could not remarkably affect osteogenic relative genes of bone trabecula and osteogenic differentiation of BMSCs.
8.Synthesis of novel beta-aminoalcohols containing nabumetone moiety with potential antidiabetic activity.
Kun ZHANG ; Jufang YAN ; Xuemei TANG ; Hongping LIU ; Li FAN ; Guangming ZHOU ; Dacheng YANG
Acta Pharmaceutica Sinica 2011;46(4):412-21
Twenty five new beta-aminoalcohols containing nabumetone moiety were prepared via the reduction of potassium borohydride with a convenient and efficient procedure, starting from beta-aminoketones that have been synthesized by our group. Their chemical structures were determined by IR, MS, 1H NMR, 13C NMR, HR-MS and antidiabetic activities were screened in vitro. Preliminary results revealed that the antidiabetic activity of most beta-aminoalcohols were better than that of the corresponding beta-aminoketones. Although most compounds showed weak antidiabetic activity, the alpha-glucosidase inhibitory activity of compounds 5hd(1) and 5id(2) reached 74.37% and 90.15%, respectively, which were superior to the positive control. The relative peroxisome proliferator-activated receptor response element (PPRE) activity of five compounds were more than 60%, among them compound 5ca possessed the highest activity (112.59%). As lead molecules of antidiabetic agents, compounds 5hd(1), 5id(2) and 5ca deserve further study.
9.Gene expression profile in osteoblastic differentiation of bone marrow stromal cells stimulated by simvastatin Gene chip analysis
Yaqiang MENG ; Liu ZHANG ; Faming TIAN ; Dacheng HAN ; Jie ZHENG ; Jun CAI
Chinese Journal of Tissue Engineering Research 2010;14(11):2081-2085
BACKGROUND:Simvastatin enhanced the expression of bone morphogenetic protein-2(BMP-2),which plays an anabolic role in bone metabolism and osteoblastic lineage differentiation.However,little is known about the molecular mechanism of simvastatin on regulation of bone marrow stromal cells differentiation.OBJECTIVE:To investigated the effect of simvastatin on osteoblastic differentiation of bone marrow stromal cells based on genetics level.METHODS:Bone marrow stromal cells derived from femur and tibia were cultured in different mediums with simvastatin or Vehicle for 7 days Following extraction and purification,mRNA was reverse-transcripted into cDNA.Fluorescence labelina was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes,which were utillzed to analyze osteogenesis-related factors.Alkaline phosphatase and Von Kossa staining were performed at days 14 and 21,respectively.RESULTS AND CONCLUSIONS:At day 14,alkaline phosphatase-positive cells were more in the experimental group than control group.Von Kossa staining demonstrated that simvastatin could promote BMSCs osteoblastic differentiation and mineralization.Comparative analysis showed that 103 genes out of 22 575 rat genes had differential expression (≥2 fold or≤ 0.5 fold),and some genes were related to cell proliferation and ostoeblastic differentiation,including C/EBP δ,Cited,Ascl2,Ptpnl6,Wisp2,Tieg,etc.Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells,involving in many osteogenetic-related genes.
10.Diagnostic value of magnetic resonance-perfusion weighted imaging in liver fibrosis of cynomolgus monkeys
Ke DING ; Xue WEI ; Manrong LIU ; Ruisui HUANG ; Shanjin LU ; Dacheng WANG ; Wei LU
Chinese Journal of Digestion 2021;41(1):43-49
Objective:To analyze the change rules of quantitative parameters of magnetic resonance-perfusion weighted imaging (MR-PWI) in cynomolgus monkeys with different degrees of liver fibrosis, and to explore the best parameter of MR-PWI in evaluating the severity of liver fibrosis.Methods:Liver fibrosis models of twenty-two cynomolgus monkeys were successfully established by subcutaneous injection of carbon tetrachloride and feeding with high-fat food. Among them, 15 cynomolgus monkeys developed into early liver cirrhosis (stage S4 of liver fibrosis). Compatibility group design was adopted, the comparative study on MR-PWI of exchange double blood supply model of liver was carried out in these 15 cynomolgus monkeys with a complete development process of liver fibrosis. The quantitative parameters of MR-PWI included endothelial transfer constant ( ktrans), reflux rate constant ( kep), extravascular extracellular space fractional volume ( ve), fractional plasma volume ( vp) and hepatic artery perfusion index (HPI). The change rules of the above parameters and their correlation with the severity of hepatic fibrosis were analyzed. The best parameter of MR-PWI was explored. Compatibility group design (randomized block design), analysis of variance, SNK- q test, Spearman rank correlation analysis and receiver operating characteristic (ROC) curve analysis were used for statistical analysis. Results:ktrans and kep of MR-PWI of cynomolgus monkeys decreased along with the progress of hepatic fibrosis, and the differences were statistically significant ( F=685.228, 99.718, both P<0.01). There were statistically significant differences between each stage of hepatic fibrosis (S1 to S4) and normal liver tissue (S0) ((0.527±0.038), (0.479±0.035), (0.432±0.032) and (0.387±0.031) mL/min vs.(0.584±0.044) mL/min, all P<0.01; (2.193±0.307), (1.997±0.301), (1.624±0.174) and (1.532±0.130) mL/min vs. (2.565±0.482) mL/min, all P<0.01). There were statistically significant in ktrans and kep between stage S3, S4 severe liver fibrosis and stage S1 mild liver fibrosis, stage S2 moderate liver fibrosis (all P<0.01), however there were no statistically significant differences between stage S3 and stage S4 liver fibrosis, between stage S1 and stage S2 liver fibrosis (all P>0.05). Along with the development of the severity of liver fibrosis, HPIs increased gradually, and the differences were statistically significant ( F=839.883, P<0.01). The HPIs of stage S0 to S4 were 0.244±0.022, 0.317±0.035, 0.421±0.046, 0.546±0.043 and 0.651±0.058, respectively, and there were statistically significant differences between groups (all P<0.01). Along with the progression of the severity of liver fibrosis, vp decreased while ve increased gradually, but there were no statistically significant differences among groups (all P>0.05). The results of Spearman rank correlation analysis indicated that ktrans and kep were negatively correlated with the severity of liver fibrosis ( rs=-0.875 and -0.797, both P<0.01), however HPI was positively correlated with the severity of liver fibrosis ( rs=0.959, P<0.01). The results of ROC curve analysis showed that area under curves (AUCs) of ktrans, kep and HPI in the diagnosis of early cirrhosis were 0.852 (95% CI 0.767 to 0.937), 0.799 (95% CI 0.700 to 0.897) and 0.967 (95% CI 0.932 to 1.002), respectively. The best cut-off values were 0.395 mL/min, 1.561 mL/min and 0.590, respectively. The sensitivity was 86.7%, 79.6% and 97.4%, respectively and the specificity was 77.4%, 71.9% and 93.1%, respectively. The thresholds of HPI in the diagnosis of liver fibrosis at stage S1, stage S2, stage S3 and stage S4 were 0.291, 0.376, 0.503 and 0.590, respectively; the sensitivity was 95.7%, 93.8% and 94.4% and 97.4%, respectively and the specificity was 89.5%, 84.7%, 91.3% and 92.7%, respectively. Conclusions:The parameters of MR-PWI change regularly with the development of liver fibrosis in the cynomolgus monkey model, among which HPI is the best parameter for quantitative evaluation of the severity of liver fibrosis.