2.Different concentrations of icariin for bone defect repair:disputes and exploration
Hongfei ZHU ; Jun ZHENG ; Xiaoyan XU ; Weizhong TANG ; Hua NIAN ; Enyuan ZHOU
Chinese Journal of Tissue Engineering Research 2014;(2):301-306
BACKGROUND:As one of the main active ingredients in epimedium, icari n plays an important role in bone defect repair. Sustained and effective concentration of icari n in vivo is essential for bone damage repair.
OBJECTIVE:To recommend the research progress of epimedium glycoside for bone repair and to explore the pharmaco-active concentration of icari n.
METHODS:A computer-based online search of CNKI database (http://www.cnki.net/) and PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) from January 2000 to October 2013 was performed for related articles of the effect of icari n for bone defect repair and bone damage repair. The key words were“icari n, concentration, bone”in Chinese and English. After repeated articles were excluded, 76 related articles were screened out and 44 of them met the inclusive criteria.
RESULTS AND CONCLUSION:The icari n-released scaffold materials can induce the osteogenic differentiation of bone marrow-derived mesenchymal stem cells, promote the viability of osteoblasts and inhibit the resorption of osteoclasts, thus repairing bone tissue. It is certain that icari n promotes cellular dif erentiation, however whether it can promote cellular proliferation remains unclear. The pharmaco-active concentration of icari n ranges from10-8 to 10-5 mol/L, but clinical trial has not yet been carried out, and specific drug concentration is uncertain, which needs further exploration.
3.Effects of coenzyme Q10 supplementation on liver mitochondrial function and aerobic capacity in adolescent athletes.
Peng LIAO ; Yong ZHANG ; Yue LIAO ; Nian-jun ZHENG ; Xu ZHANG
Chinese Journal of Applied Physiology 2007;23(4):491-494
AIMTo investigate the effects of CoQlo supplementation on liver mitochondrial function and aerobic capacity in adolescent athletes.
METHODSBased on a single blinded study design, 18 male adolescent swimming athletes were randomized into two groups, supplement CoQ10 100 mg/d (Q group), or placebo (P group) for 28 days respectively.
RESULTS(1) After supplementation, the plasma CoQ10 concentration in Q group was significantly elevated and significantly higher compared to P group. (2) After supplementation, the rest plasma MDA level in Q group remained unchanged and was significantly lower compared to P group. (3) The plasma CoQ10 concentration of the 18 athletes was significantly decreased during the first constant endurance exercise. (4) The baseline plasma CoQ10 of the 18 subjects showed significantly positive correlation with VO2max measured in the first incremental exercise. (5) No significant difference of increased level of AKBR between Q group and P group. (6) No significant difference of increase level of VO2max, individual lactate threshold and exercise economy between Q and P group.
CONCLUSIONAlthough there is an increased demand for plasma CoQ10 during endurance exercise and CoQ10 supplement can depress lipid peroxidation, there is no effect of CoQ10 supplementation on liver mitochondrial function and aerobic capacity in adolescent athletes.
Adolescent ; Exercise ; Humans ; Lipid Peroxidation ; Male ; Mitochondria, Liver ; metabolism ; physiology ; Physical Endurance ; Swimming ; Ubiquinone ; administration & dosage ; analogs & derivatives
4.Activation of transcription factors induced by low concentration of N-methyl-N'-nitro-N-nitrosoguanidine.
Gu-liang WANG ; Zheng WANG ; Jun YANG ; Ying-nian YU
Journal of Zhejiang University. Medical sciences 2003;32(5):390-392
OBJECTIVETo investigate the effect of MNNG on some of the transcription factors such as NF- kappaB, CREB, AP-1 and c-Myc.
METHODSThe activities of these transcription factors were measured by transient transfection assay of SEAP vectors.
RESULTThe expressions of AP-1, CREB and NF- kappaB driven reporter genes were elevated for about 1.3, 1.4 and 1.3 times in MNNG-treated cells, respectively, as compared to untreated controls. However, the exposure of MNNG had no effect on the activity of c-Myc.
CONCLUSIONThe activation of certain transcription factors might be involved in the process of untargeted mutation induced by low concentration of MNNG treatment.
Animals ; Cercopithecus aethiops ; Cyclic AMP Response Element-Binding Protein ; drug effects ; Methylnitronitrosoguanidine ; toxicity ; Mutation ; NF-kappa B ; drug effects ; Proto-Oncogene Proteins c-myc ; drug effects ; Transcription Factor AP-1 ; drug effects ; Transcription Factors ; drug effects ; Vero Cells
5.Clinical application of percutaneous endoscopic gastrostomy in gastrointestinal nutrition among critically ill pa-tients
Hua LIN ; Rui-Qiang ZHENG ; Hai-Hang ZHU ; Nian-Fang LU ; Qi-Hong CHEN ; Jun SHAO ; Jiang-Quan YU ;
Chinese Journal of Primary Medicine and Pharmacy 2005;0(11):-
Objective To explore the clinical application of percutaneous endoscopic gastrostomy in gastroin- testinal nutrition among critically ill patients.Methods Twenty-two patients were undertaken percutaneous endo- scopic gastrostomy under the lead of gastroscope.The puncturatio site was located in anterior wall of stomach.Results All the procedures were performed successfully for one time.Intraoperational blood pressure was very steady.At the same time,intraoperational SpO_2 of the patients all exceeced 97%.The bleeding amount and operation time were respectively (3.8?1.9) ml and (15.5?2.3) min.Severe complications such as gastrostoma,gastrocolic fistula, pneumoperitoneum,refluxing or aspiration of gastric juice,inhalant pneumonia didn't occur after the operation.Con- clusion Percutaneous endoscopic gastrostomy was safe and feasible among critically ill patients who needed gastroin- testinal nutrition because of its slight injury,little bleeding and shorter operational time.
6.Ocular inflammation and pathological characteristics of recurrent experimental autoimmune uveitis in rat
Hui, ZHENG ; Xiao-min, ZHANG ; Hong, NIAN ; Ling-jun, ZHANG ; Xun, LIU ; Shao, HUI ; Xiao-rong, LI
Chinese Journal of Experimental Ophthalmology 2013;(7):642-646
Background Most animal models of experimental autoimmune uveitis (EAU) are single attacked procedure,with a different feature from the natural course of human recurrent autoimmune uveitis.So establishing a recurrent EAU model is necessary for the clinical study on EAU.Objective This study was to establish the recurrent EAU model in rat and investigate the ocular inflammation and pathological manifestation and interleukin-17 (IL-17)expression in the eye.Methods T cells isolated from the spleen and draining lymph nodes of Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide fragments (R16) 10 days earlier were re-stimulated with R16 in vitro and injected into naive syngeneic rats to establish the recurrent EAU models,and the normal Lewis rats were used as controls.The eyes of model rats were then examined daily for clinical signs of uveitis by slit-lamp biomicroscopy and scored Caspi's criteria.The rats were sacrificed 1 month,2,3months after injection respectively,and the retinal sections were prepared for the pathological examination by hemotoxylin & eosin staining.Immunohistochemistry was performed to detect the expression of IL-17 in the retina.Results Adoptive transfer of R16-specific T cells to Lewis rats induced recurrent uveitis.The inflammatory scores on the fourth day,the sixth day,and the inflammatory response disappeared on the tenth day after injection.However,the inflammatory reaction occurred repeatedly 4 or 5 times in the 2-month duration after that,and the right and left eyes of a single recipient showed a different pattern of relapse,and the clinical manifestations of EAU was similar to the natural course to those of human autoimmune uveitis.In the retinal specimens of 1-,2-and 3-month group,the number of inflammatory cells was gradually decreased as the time lapse.Compared with the normal group,the thicknesses of the entire retina,outer nuclear layer and inner nuclear layer decreased with a significant difference among the 4 groups (F=20.46,288.40,4.43,all P=0.00).The number of RGCs in the normal group,1-,2-and 3-month group was 231.27 ± 15.36,225.36 ± 17.79,132.18 ±9.39 and 67.45 ± 11.90,respectively,showing a significant difference among them (F=68.94,P=0.00).Immunohistochemistry showed that the scores of the IL-17 expression in the rat retina were 0.64 ± 0.17,1.92 ± 0.19,1.17 ± 0.23 and 0.83 ± 0.23,showing statistically significant difference (F=64.10,P=0.00).Conclusions The stimulation of R16-specific T cells can induce recurrent EAU in Lewis rat.Th17 is involved in the disease course.
7.Effect of Yangjing Zhongyu Decoction on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulosa cells cultured by excess androgen.
Yan-Hua ZHENG ; Tao DING ; Hong-Xia MA ; Dan-Feng YE ; Nian-Jun SU ; Xiao-Ke WU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(3):312-316
OBJECTIVETo observe the effect of Yangjing Zhongyu Decoction (YZD) on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulose cells (GCs) cultured by excess androgen.
METHODSOvarian GCs from porcine follicles were isolated and cultured in vitro. Follicular stimulating hormone (FSH) or YZD was added in the GCs treated by excess testosterone propionate. Totally 48 h later mRNA and protein expression of PCNA, StAR, and FSHR were detected by RT-PCR and Western blot.
RESULTSExcess androgen inhibited mRNA and protein expression of PCNA, StAR, and FSHR of GCs. FSH and YZD could antagonize inhibition of excess androgens, and promote mRNA and protein expression of PCNA, StAR, and FSHR in GCs.
CONCLUSIONYZD could antagonize the inhibition of excess androgen on mRNA and protein expression of PCNA, StAR and FSHR in GCs. Thus, we inferred that YZD could improve the follicle dysplasia by promoting mRNA and protein expression of PCNA, StAR and FSHR in GCs.
Androgens ; pharmacology ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Membrane Transport Proteins ; genetics ; metabolism ; Ovarian Follicle ; cytology ; drug effects ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, FSH ; genetics ; metabolism ; Swine
8.Construction of an oncolytic adenovirus expressing small hairpin RNA and targeting the SATB1 gene.
Li-Jun MAO ; Jun-Nian ZHENG ; Wang LI ; Jun-Qi WANG ; Jia-Cun CHEN ; Xiao-Qing SUN
National Journal of Andrology 2010;16(8):679-683
OBJECTIVETo construct an oncolytic adenovirus with the DD3 promoter regulation, expressing small hairpin RNA and targeting the SATB1 gene (SATBI-shRNA), and to evaluate its potential for inhibiting the growth of human prostatic carcinoma cells (LNCaP) in vitro.
METHODSSATB1-shRNA expression cassettes containing the H1 promoter were produced by PCR from pSilencer3. 1-SATB1 and inserted into the pZD55 vector, and the recombinant plasmid pZD55-SATB1-shRNA was constructed, pZD55SATB1-shRNA and pZXC2-DD3-E1A were digested with EcoRV and Xba I , and the obtained expression cassettes linked each other to construct the recombinant plasmid pDD3-ZD55-SATB1, which was cotransfected with the pBHGE3 packaging plasmids mixture into 293 cells by Effectence. The recombined adenoviruses DD3-ZD55-SATB1 were identified by PCR. Viruses were propagated on HEK293 cells and purified by standard techniques, and the functional PFU titers determined by plaque assay on 293 cells. The antitumor potential of DD3-ZD55-SATB1 to LNCaP was evaluated by the crystal violet dye method. The expression level of the E1A gene was detected by Western blot, and that of the SATB1 gene in the adenovirus-infected LNCaP cells by both Western blot and RT-PCR.
RESULTSAn oncolytic adenovirus expressing SATB1-shRNA with the DD3 promoter regulation, DD3-ZD55-SATB1, was constructed successfully, whose functional PFU titer was 1.2 x 10(10) PFU/ml. DD3-ZD55-SATB1 showed an obvious cytopathic effect and a selective expression of E1A in the adenovirus-infected LNCaP cells; it exhibited a high LNCaP-targetability and specific SATB1-silencing effect.
CONCLUSIONThe successful construction of the oncolytic adenovirus DD3-ZD55-SATB1 offers a new tool for researches on the gene therapy for human prostate cancer.
Adenoviridae ; genetics ; Carcinoma ; therapy ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Male ; Matrix Attachment Region Binding Proteins ; genetics ; Oncolytic Virotherapy ; methods ; Oncolytic Viruses ; genetics ; Promoter Regions, Genetic ; Prostatic Neoplasms ; therapy ; RNA Interference ; RNA, Small Interfering ; genetics
9.Effects of adenovirus-mediated human bone morphogenetic protein-7 on proliferation and differentiation of human dental pulp cells.
Zheng-mei LIN ; Wei QIN ; Jun-qi LING ; Nian-hua ZHANG ; Lin XIAO
Chinese Journal of Stomatology 2006;41(10):612-615
OBJECTIVETo investigate the effect of adenovirus expressing human bone morphogenetic protein-7 (hBMP-7) on proliferation and differentiation of human dental pulp cells.
METHODSThe replication-deficient adenoviral vector encoding hBMP-7 gene was constructed by using homologous recombinant modality. The efficiency of transfection was evaluated by fluorescent microscopy and flow cytometry. The expression of hBMP-7 protein in adenovirus-infected dental pulp cells was determined by Western blot. The proliferation of cells was tested by MTT method, the activity of alkaline phosphatase was assayed, von Kossa staining was used to detect mineralized nodule formation, and the expression of DSPPmRNA in cells was detected using semi-quantitative RT-PCR.
RESULTSGreen fluorescent protein was visible under fluorescent microscopy. Higher transfection efficiency (91.1 +/- 1.0)% could be obtained at MOI of 75. Western blot from dental pulp cells infected with Ad-hBMP-7 for 48h detected protein expression of a hBMP-7 gene. The activity of alkaline phosphatase in cells was significantly higher than those of the control groups (P < 0.05). The cells infected with Ad-hBMP-7 had the ability of mineralization. DSPP mRNA expression of cells was in a time- and dose- dependent manner.
CONCLUSIONSAd-hBMP-7 can induce human pulp cells into odontoblasts, but has no obvious effect on their proliferation.
Adenoviridae ; genetics ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Dental Pulp ; cytology ; Humans ; Odontoblasts ; cytology ; Transfection
10.Activation of nucleus-independent signals triggered by N-methyl-N'-nitro-N- nitrosoguanidine.
Zheng WANG ; Gu-liang WANG ; Jun YANG ; Zhi-hua GAO ; Ying-nian YU
Journal of Zhejiang University. Medical sciences 2003;32(5):385-389
OBJECTIVETo study the effect of MNNG on inducement of non-targeted mutation and activation of several cellular signal transduction pathways, and to determine whether the activation of these signaling pathways was dependent on the DNA-damage.
METHODSVero cells were enucleated by discontinuous density centrifugation. The PKA activities were measured by enzyme-linked immunosorbent assay. The status of cell membrane receptors was studied with immunofluorescent staining and confocal microscopy.
RESULTIn enucleated cytoplasts, MNNG-treatment increased PKA activity for about 2.3-fold in accordance with the 2.7-fold up-regulation of PKA activity in whole vero cells exposed to MNNG. The clustering of cell surface receptors of epidermal growth factor and tumor necrosis factor alpha was also observed in cells exposed to MNNG; this phenomenon was also found in enucleated cells.
CONCLUSIONThe results indicate that the initiation of signal cascades induced by low concentration of MNNG might be associated with its interaction with cell surface receptors and/or direct activation of related signal proteins but not its DNA damage.
Animals ; Cell Nucleus ; physiology ; Cercopithecus aethiops ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; DNA Damage ; Enzyme Activation ; drug effects ; Methylnitronitrosoguanidine ; toxicity ; Receptor, Epidermal Growth Factor ; drug effects ; metabolism ; Receptors, Tumor Necrosis Factor ; drug effects ; metabolism ; Signal Transduction ; drug effects ; Vero Cells