2.Cellular Apoptosis of C6 Mouse Glioma Cells Induced by hING4 Mediated by Adenovirus
Yao-Dong ZHAO ; Jing-Cheng MIAO ; Hai-Feng ZHANG ; Wei-Hu SHENG ; Li MIAO ; Yu-Feng XIE ; Ji-Cheng YANG ;
China Biotechnology 2006;0(07):-
The known members of inhibitor of growth (ING) gene family are considered as candidate tumor suppressor genes. ING4, a novel member of ING family, is recently reported to regulate brain tumour angiogenesis through transcriptional repression of NF-?B-responsive genes, induce G2/M arrest by the increased p21 expression in a p53-dependent manner, suppress the loss of contact inhibition and represses activation of the hypoxia inducible factor, which plays an important role in the progression of tumorigenesis. However, seldom studies about ING4 inducing tumor cells apoptosis were reported.The C6 cells (mouse glioma cells) were infected respectively with the blank adenovirus carrying GFP (Ad) and the recombinated Ad-hING4-His, then RT-PCR assay was used to detect the transcriptions of hING4, as well Western-blotting assay was ued to detect the expressions of hING4. The effects of hING4 expression upon C6 cells were observed, and the growth curve was drawed and tumor control rates were calculated. The C6 cells, which were affected by blank Ad and Ad-hING4-His, were respectively observed by LSCM (laser scan confocal microscope) and transmission electron microscope (TEM), detected by flow cytometry; and the genomic DNA of both groups were extracted and electrophoresised in agarose gel to examinate the DNA fragments. The results showed hING4 can significantly inhibit the growth of C6 cells by promoting the cell’s apoptosis, which probably is the first one to prove this property of ING4.The experimental and theoretical foundation for gene therapy for gliomas with ING4 in the future was established.
3.Chromosomal localization of primary gout susceptibility gene in a Chinese pedigree
Chang-Gui LI ; Ying CHEN ; Chao XU ; Zhi-Min MIAO ; Sheng-Li YAN ; Huai-Dong SONG
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To determine chromosomal localization of the primary gout susceptibility gene in a pedigree.Methods The clinical data and the peripheral blood samples were collected in the pedigree members and the genomic DNA was extracted from peripheral blood.A genome-wide screening was performed using 400 micro-satellite DNA markers in this family,and linkage analysis was used to determine the chromosomal location of the primary gout susceptibility gene.Results Linkage analysis showed that the maximum LOD score reached 1.50 at marker D4S1572 (at recombination fraction?=0.00).Conclusion Since D4S1572 is localized at 4q25,the primary gout susceptibility gene of this pedigree is localized at 4q25.
4.Effects of auricular acupuncture on the memory and the expression of ChAT and GFAP in model rats with Alzheimer's disease.
Ting MIAO ; Tian-sheng JIANG ; Yu-hua DONG ; Nai-chang JIANG
Chinese Acupuncture & Moxibustion 2009;29(10):827-832
OBJECTIVETo observe the effects of auricular acupuncture on the learning and memory abilities of model rats with Alzheimer's disease (AD), and investigate its mechanism.
METHODSThirty SD rats were randomly divided into a control group, a model group and an auricular acupuncture group, 10 rats in each group. The model rats with AD were established by multiple injections with Okadaic Acid into the CA1 region of hippocampus. In the control group, the same quantity injection with Dimethyl Sulfoxide (DMSO) was applied on experimental rats. The auricular acupoints of "Nao" (brain) and "Shen" (kidney) were used for treating in the auricular acupuncture group, in contrast, the auricular region were not treated in the model and the control groups. The learning and memory capabilities of the rats were assessed with Morris Water Maze behavioral test, and the expressions of choline acetyltransferase (ChAT) and glial fibrillary acidic protein (GFAP) were examined by immunohistochemistry.
RESULTSComparing with the model group, the treated AD rats with auricular acupuncture was showed that the average escape latency was obviously shortened in the place navigation test (P<0.01), the movement time in plateform quadrant was obviously prolonged in the spatial probe test (P<0.05), and the number of traversing platform obviously increased (P<0.01) after the platform was taken away. The expression of ChAT increased in the hippocampus and cortex (P<0.01, P<0.05), but the expression of GFAP obviously decreased in the CA1 region of hippocampus (P<0.01).
CONCLUSIONAuricular acupuncture can improve the learning and memory capability of the model rats with AD. Its mechanism might be related with decreasing cholinergic neuron damage and reducing the abnormal activation and hyperplasia of astrocyte.
Acupuncture, Ear ; Alzheimer Disease ; genetics ; metabolism ; psychology ; therapy ; Animals ; Choline O-Acetyltransferase ; genetics ; metabolism ; Disease Models, Animal ; Gene Expression ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Humans ; Male ; Memory ; Random Allocation ; Rats ; Rats, Sprague-Dawley
5.Effect of curcumine on the expression of Fas/FasL in rat brain tissue under chronic low O2 and high CO2.
Jun-Li LI ; Yan-Yan FAN ; Guang-Hua YE ; Miao-Wu DONG ; Ke-Zhi LIN ; Feng LI ; Lin-Sheng YU
Chinese Journal of Applied Physiology 2014;30(2):165-167
Animals
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Brain
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drug effects
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metabolism
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Carbon Dioxide
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metabolism
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Chronic Disease
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Curcumin
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pharmacology
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Disease Models, Animal
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Fas Ligand Protein
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metabolism
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Hypoxia
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metabolism
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Male
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Oxygen
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metabolism
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Rats
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Rats, Sprague-Dawley
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fas Receptor
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metabolism
6.Case-control study on suture-assisted locking plate for the treatment of proximal humeral fractures in elderly.
Zhang-sheng DAI ; Jie-miao HUANG ; Xun-rong ZHUANG ; Shou-bo CHEN ; Shi-qiang WU ; Xue-dong YAO ; Hui YE
China Journal of Orthopaedics and Traumatology 2014;27(12):1015-1018
OBJECTIVETo explore clinical effects of suturing-assisted locking plate in treating elderly proximal humeral fractures.
METHODSFrom January 2005 to January 2013, 55 elderly patients with three- and four-part fractures of proximal humeral fractures were divided into treatment group and control group. In treatment group, there were 31 patients including 12 males, and 19 females aged from 65 to 85 with an average of (74.00±5.42) years old, and treated with suturing-assisted locking plates; 19 patients were Neer 3-part fractures, and 12 patients were Neer 4-part fractures of proximal humerus; 23 patients were suffered from low-energy injuries and 8 patients were caused by high-energy injuries. In control group, there were 24 patients including 7 males, and 17 females aged from 65 to 85 with an average of (72.79±5.34) years old, and treated with locking plates; 16 patients were Neer 3-part fractures, and 8 patients were Neer 4-part fractures of proximal humerus; 17 patients were suffered from low-energy injuries and 7 patients were caused by high-energy injuries. Operative time, blood loss during operation, and bone healing time between two groups were observed and compared. Postoperative Neer scoring were used to evaluate recovery of shoulder joint function.
RESULTSAll patients were followed up from 6 to 24 months with an average of 16.1 months. In treatment group, blood loss was (495.806±143.150) ml, function of Neer scoring was 22.645±2.443, range of action was 18.194±2.613, anatomy was 7.935±1.504 and total score of Neer scoring was 77.161±8.335; while in control group, blood loss was (641.667±169.851) ml, function of Neer scoring was 13.958±1.989, range of action was 13.083±2.165, anatomy was 5.500±1.978 and total score of Neer scoring was 58.792±7.313. There were sigificant difference between two groups in these indexes.
CONCLUSIONSuturing-assisted locking plate for the treatment of proximal humerus fractures in elderly, has advantages of less blood loss, simple fracture reduction and rapid recovery of shoulder joint, and is a effective method.
Aged ; Aged, 80 and over ; Bone Plates ; Case-Control Studies ; Female ; Humans ; Male ; Recovery of Function ; Shoulder Fractures ; physiopathology ; surgery ; Shoulder Joint ; physiopathology ; Sutures
7.Expression of Human Interferon-?1 and Interferon-? Gene in WI-38 Cells and Comparison of Their Biological Activity
Bhushan SARODE ; Wei-Hua SHENG ; Yu-Feng XIE ; Yao-Dong ZHAO ; Jing-Cheng MIAO ; Ji-Cheng YANG ;
China Biotechnology 2006;0(03):-
The biological!activities i.e. antineoplastic activities and antiviral activity of the two novel kinds of interferons: hIFN-?1 and hIFN-? were studied and compared. First the fusion expression vector: pcDNA3.1A-hIFN-?1-His and pcDNA3.1A-hIFN-?-His by PCR was constructed,then the two kinds of plasmids were transfected into WI-38 (human embryonic lung cells) with liposome. And cytopathic effect (CPE) suppression test was used to study and compare the antiviral activities of rhIFN-?1-His and rhIFN-?-His, meanwhile MTT assay was used to detect their antineoplastic activities.It was found that, antiproliferative activity and MxA protein induction shown by rhIFN-?1-His is more powerful than of rhIFN-?-His. The antiviral molecular mechanisms of both hIFN-?1 and hIFN-? are related to MxA.The foundation for further study on the bioactivities and mechanism of action of hIFN-?1 and hIFN-? was established.
8.Effect of graded composite zirconia-hydroxyapatite on viability of rat osteoblast cells cultured in vitro
Ren-Fu QUAN ; Di-Sheng YANG ; Xu-Dong MIAO ; Wei LI ; Xiao-Chun WU ; Hong-Bin WANG
Chinese Journal of Trauma 2003;0(11):-
Objective To evaluate the effect of a novel orthopedic biomaterial,graded composite zireonia(ZrO2)hydroxyapatite(HAP)on activity of rat osteoblast ceils(OB)cultured in vitro. Methods The pure zirconia material was used as control to measure surface roughness of the composite material that was examined by using scanning electron microscope(SEM)and X-ray diffraetometer (XRD).The rat osteoblast cells were cultured on the two materials.Alkali phosphatase(ALP)of the two groups was measured and ELISA was used to detect IL-6 and TGF-?eoncentration of the supematant of OB cells.Tumor growth factor-?(TGF-?)mRNA was detected by RT-PCR.SEM was used to observe OB cells on the two materials.The extract of the composite material was used for a eytotoxicity test to cal- culate the relative proliferation rate(RGR)and classify the toxicity.Results The surface roughness of the gradual composite materials was significantly higher than that of the control materials(P<0.01). The ALP of the gradual material group was markedly higher than that of the control group at different in- tervals.There was significant difference of the IL-6 and TGF-?concentrations 2-4 days after culture be- tween two groups(P<0.05,P<0.01).The mRNA level of TGF-?of the two OB groups also showed marked statistical difference(P<0.01).The ossification of the OB cells on the composed material was marked after 14 days.The MTT color experiments showed no statistic significance between materials group and negative group,with the toxicity at levelⅠand 0(P<0.05).Conclusion Graded composite ZrO2 HAP can significantly promote proliferation and differentiation of OB cells cultured in vitro and has good biocompatibility.
9.Synergistic induction of apoptosis by the combination of TRAIL and low dose adriamycin in human osteosarcoma cell line U2OS.
Di-sheng YANG ; Xu-dong MIAO ; Zhao-ming YE ; Yu-sheng XU
Chinese Journal of Oncology 2005;27(10):595-597
OBJECTIVETo investigate whether TRAIL can synergize with adriamycin (ADM) to kill osteosarcoma cells (U2OS) in vitro, and its possible molecular mechanism.
METHODSMTT was used to evaluate the cytotoxic effect of TRAIL and ADM either used alone or in combination at 24 hours after treatment to U20S cells. Cell apoptosis and its proportion were detected by flow cytometry assay. Acridine orange fluorescence microscopy and transmission electron microscopy were used to examine cellular and ultrastructural changes of apoptosis. The changes of cFLIP in mRNA and protein level were semi-quantified by RT-PCR and Western blot analysis.
RESULTS(1) U2OS cells were not sensitive to TRAIL (IC(50) > 1 mg/L); the cells were relatively more responsive to ADM in an apparent dose-effect fashion. (2) The combination of TRAIL and ADM presented a synergistic effect on U2OS cells. Subtoxic concentration of TRAIL (0.1 mg/L) combined with subtoxic concentration of ADM (1.0 micromol/l) killed (49.54 +/- 2.79)% of U2OS cells. (3) The cytotoxicity was mainly attributed to cell apoptosis as demonstrated by flow cytometry assay, fluorescence microscopy and electron microscopy.
CONCLUSIONSubtoxic dose of TRAIL can effectively kill osteosarcoma cells (U2OS) in combination with subtoxic dose of ADM, but not effective when used alone. Apoptosis is the main mechanism of this killing effect induced by combination of TRAIL and ADM. Down regulation of cFLIP at mRNA and protein level is involved in this apoptosis pathway.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Bone Neoplasms ; pathology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; Caspase Inhibitors ; Doxorubicin ; pharmacology ; Drug Synergism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Membrane Glycoproteins ; pharmacology ; Osteosarcoma ; pathology ; RNA, Messenger ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; pharmacology
10.Effect of apigenin on the vasodilatory of basilar artery in rats and it mechanism
Peng-Mei GUO ; Yu LIU ; Yi-Xin JING ; Qi-Ying SONG ; Miao-Miao DONG ; Li-Na DONG ; Ming-Sheng ZHANG
The Chinese Journal of Clinical Pharmacology 2018;34(9):1100-1104
Objective To study the spasmolytic effect and the underlying mechanism of apigenin in isolated rat basilar artery.Methods Healthy male rats were euthanized by exsanguination.The rat brain was removed and transferred immediately into physiological salt solution.The rat basilar arteries were isolated and cut into 2 mm-long rings under the microscope.Diastolic test:the different segments of the same blood vessel were randomly divided into two groups:the control group (solvent dimethylsulfoxide,diastolic) and the experimental group (apigenin 1-100 μmol · L-1,diastolic).Inhibition of contraction test:the same blood vessel was divided into the control group,low,medium and high dose experimental groups based on the dosage of administration (apigenin 0,10,30,100 μmol · L-1 respectively).The duration of administration was at least 10 min.Calcium ion (Ca2+) deprivation and specific inhibitors:an inhibitor of protein kinase C (Go6983),an inhibitor of extracellular signal-regulated kinase (PD98059),an inhibitor of Rho-associated kinase (Y-27632),an inhibitor of p38 Mitogen-activated protein kinase (SB239063) were used to explore the possible involvement of modulation of Ca2 + availability and activities of protein kinases in apigenin-induced vasorelaxation.The vascular myogenic tone was recorded with a wire myograph.Half maximal diastolic concentration (RC50),half maximal inhibitory concentration (IC50),maximal diastolic rate and maximal inhibition rate were calculated.Results After the administration,apigenin 1 ~ 100 μmol · L-1 concentration-dependently relaxed the arterial rings precontracted with either KCl 60 mmol · L-1,U46619 0.3 μmol · L-1 or vasopressin (VP,3 μmol · L-1) and the RC50 values were (24.27 ±1.14),(4.1 ±1.16),(1.9±1.33) μmol · L-1 respectively.Comparison between experimental groups with control group,the difference had significantly (all P < 0.01).Incubation with apigenin 10 ~ 100 μmol · L-1 shifted the concentration-contraction curves of KCl,U46619,VP or Ca2+ to the right with the maximal contractions depressed.The IC50values of low,medium and high dose experimental groups against KCl,U46619,VP or Ca2+ were (77.73 ± 1.72),(14.91 ± 1.31),(20.66 ± 1.21),(7.53 ± 1.02) μmol · L-1 respectively.Comparison between three doses experimental groups with control group,the difference had significantly (all P < 0.01).Both intracellular Ca2+ release dependent and extracellular Ca2+ influx dependent contractions were reduced after preincubation with apigenin by (24.53 ± 5.32) % and (48.58 ± 8.92) %.Preincubation with Go6983 and PD98059,but not Y-27632 and SB239063 attenuated apigenin-induced vasorelaxation.Relaxation rate of Go6983 made API on the KCl,U46619 shrinkingwere decreased (40.33 ± 2.51)%,(36.51±5.13)%;Relaxation rate of PD98059 made API on the KCl,U46619 shrinkingwere decreased (25.84 ± 3.86)%,(21.80 ± 5.95)%.Compared with control group (without apigenin or inhibitors intervention),the difference were statistically significant (P <0.05,P <0.01).Conclusion Apigenin are spasmolytic against a variety of vasoconstrictors in rat basilar artery.Reduction of Ca2 + availability,modulation of activities of extracellular signal-regulated kinase and protein kinase C may be,at least in part,contributory to vasorelaxant effects of apigenin.