1.The co-stimulatory effect of growth factors on chondrogenesis induction of rat mesenchymai stem cells
Feng-Ying GAN ; Chen TANG ; De-Fu YE ; Di-Bin GUO ;
Chinese Journal of Rheumatology 2003;0(11):-
Objective The present study aims to explore the optimal combination of growth factors, including TGF-?_1,IGF-I and BMP-2,for the induction of ehondrogenesis of MSCs in vitro monolayer culture. Methods MSCs were induced to differentiate into ehondrocytes in the medium with the following growth fac- tors:TGF-?_1,BMP-2 and IGF-I,either alone or in combination.We detected two cartilage specific markers in differentiated cells.Alcian blue stain was used for aggreean and RT-PCR for collagen type II mRNA.Results The level of Collagen typeⅡmRNA of group BMP-2+IGF-I,TGF-?_1,TGF-?_1+IGF-I,TGF-?_1+BMP-2 and TGF-?_1+IGF-I+BMP-2 was 0.147?0.062,0.478?0.069,0.489?0.022,0.653?0.013,0.807?0.051 respectively. Alcian blue stain in all gruups were positive.Conclusion The TGF-?_1+IGF-I+BMP-2 group is the most ef- fective combination in chondrogenesis induction.
2.Construction and transplantation of tissue-engineered skin with mouse embryonic fibroblasts in SD mice.
Liu LIU ; Wu-de LI ; Guo-bin CAI
Chinese Journal of Plastic Surgery 2011;27(4):284-289
OBJECTIVETo investigate the application and mechanism of tissue-engineered skin with mouse embryonic fibroblasts (MEFs) for the full-thickness skin defects on mice.
METHODSThe MEFs and fibroblasts were cultured and seeded in scaffold made of rat tail collage. ELISA method was used for detection of secretory function. The full-thickness skin defects were created on mice and covered by MEFs-scaffold complex (experimental group), or FBs-scaffold complex (control group 1), or scaffold only (control group 2). The process of wound healing was evaluated by observation of the re-epithelization rate. Microvessel density (MVD) and vimentin within the wound sites were also detected with immunohistochemistry staining technique to describe the characteristics of wound healing. Hoechst 33342 staining was performed to trace MEFs'fate.
RESULTSMEFs scaffold group had higher level secretion of IL-6 and lower of TGF-beta1 than FBs scaffold group (P<0.05). Compared with wounds in control groups, the wounds in MEFs group healed markedly fast (P<0.05) and the MVD was significantly higher (P <0.05). The fibroblasts in the wounds of MEFs group were arranged regularly and the MEFs decreased during the healing process.
CONCLUSIONSThe MEFs-scaffold complex can promote wound healing with less scar formation. MEFs may have an inducing effect on the wound healing.
Animals ; Female ; Fibroblasts ; cytology ; Interleukin-6 ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Skin, Artificial ; Tissue Engineering ; Tissue Scaffolds ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing
3.The inhibitory effect of Euonymus alatus against mouse hepatic fibrosis induced by CCl4
Xing WAN ; Qiong GUO ; Xiang-Dong LIU ; De-Bin HUANG
Chinese Pharmacological Bulletin 2018;34(4):485-490
Aim To explore the inhibitory effect of Euonymus alatus on hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice and its mechanism. Methods Eighty C57BL/6 male mice were randomly divided into eight groups: normal group, CCl4model group, Euonymus alatus(EA) ethanol extracts groups in early stage(EAE), EA ethanol extracts groups in later stage(EAL),two drug groups with low/medium/high dose(EAE-L/M/H, EAL-L/M/H), with 10 mice in each group. Fibrosis model was established by injecting CCl4in peritoneal cavity,and the study lasted for 30 days. Different doses of drugs were given from 1 st day to 15 th day in EAE while from 16 th day to 30 th day in EAL,then all mice were sacrificed to for the observation of the morphological changes and collage-nous fiber by HE and Masson staining. Liver index, ALT,AST and TNF-α were tested by ELISA. The ex-pressions of α-SMA and CollagenⅠwere measured by immunohistochemistry and Western blot. Results Compared to normal group, liver index, ALT, AST, TNF-α, α-SMA and CollagenⅠ in EA groups were lower than those in model group in a dose-dependent manner(P<0.01 or P<0.05). Liver morphology and collagenous fiber in EAE and EAL were better than those in model group in a dose-dependent manner. The effect of EAE were superior to that of the EAL in HE, Masson, α-SMA, Collagen Ⅰ indexes(P <0.05). Conclusions Euonymus alatus may inhibit the process of hepatic fibrosis in mice with dose-effect de-pendence, and drug treatment in early stage performs better,which may be related to the decrease of TNF-α that affects the expression of α-SMA and Collagen Ⅰ.
4.To investigate plasma levels of free and total carnitine in patients undergoing hemodialysis or peritoneal dialysis
Li-Li JIAO ; Fu-De ZHOU ; Guo-Bin XU ; Xue-Jing WANG ; Jie DONG ; Hai-Xia LI ; Tie-An XIA ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.
5.Misdiagnosis analysis of tumor-induced hypophosphatemic osteomalacia: a report of 3 cases.
China Journal of Orthopaedics and Traumatology 2010;23(3):230-232
Adult
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Diagnostic Errors
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Humans
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Hypophosphatemia
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diagnosis
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etiology
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therapy
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Male
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Middle Aged
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Neoplasms
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complications
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Osteomalacia
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diagnosis
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etiology
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therapy
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Young Adult
6.Reply: The Characteristics about Our Study in Irritable Bowel Syndrome.
Kang Min ZHUANG ; Yu Bin GUO ; Si De LIU
Gut and Liver 2017;11(1):168-168
No abstract available.
Irritable Bowel Syndrome*
7.Palmitic acid induces hepatocellular oxidative stress and activation of inflammasomes.
Wen XU ; Yu-Bin GUO ; Xu LI ; Mei-Rong HE ; Si-de LIU
Journal of Southern Medical University 2016;36(5):655-659
OBJECTIVETo evaluate the effect of palmitic acid (PA) on oxidative stress and activation of inflammasomes in hepatocytes.
METHODSTo test the dose-dependent effect of PA on normal murine hepatocytes AML12, the cells were treated with 0, 0.15, 0.25 and 0.4 mmol/L of palmitic acid (PA). The cells were also divided into blank control group, 0.25 mmol/L PA group and 0.25 mmol/L PA+N-acetylcysteine (NAC) group to examine the effect of reactive oxygen species (ROS) on the activation of inflammasomes. After 24 h of treatment, lipid accumulation, total ROS, mitochondrial ROS, expression and localization of NOX4, and expressions of inflammasomes and IL-1β were detected in the hepatocytes.
RESULTSCompared with the control cells, PA treatment of the cells significantly increased cytoplasmic lipid accumulation, concentrations of total ROS (12 463.09±2.72 vs 6691.23±2.45, P=0.00) and mitochondrial ROS (64.98±0.94 vs 45.04±0.92, P=0.00), and the expressions of NOX4, NLRP3, ASC, caspase-1, and IL-1β (1603.52±1.32 vs 2629.33±2.57, P=0.00). The mitochondria and NOX4 were found to be co-localized in the cytoplasm. NAC obviously reduced cellular ROS level stimulated by PA (7782.15±2.87 vs 5445.6±1.17, P=0.00) and suppressed the expressions of NLRP3, ASC and caspase-1.
CONCLUSIONPA treatment can stimulate lipid accumulation in hepatocytes and induce oxidative stress through NOX4 and mitochondria pathway to activate inflammasomes and stimulate the secretion of IL-1β.
Acetylcysteine ; pharmacology ; Animals ; Carrier Proteins ; metabolism ; Caspase 1 ; metabolism ; Cells, Cultured ; Hepatocytes ; drug effects ; metabolism ; Inflammasomes ; drug effects ; metabolism ; Interleukin-1beta ; metabolism ; Mice ; Mitochondria ; drug effects ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein ; Oxidative Stress ; Palmitic Acid ; pharmacology ; Reactive Oxygen Species ; metabolism
8.Construction and expression of anti-tumor necrosis factor related apoptosis-inducing ligand receptor death receptor 5 chimeric antibody in eukaryotic cells.
Feng CHEN ; Ya-Bin GUO ; Shi-Lian LIU ; De-Xian ZHENG ; Yan-Xin LIU
Acta Academiae Medicinae Sinicae 2008;30(6):690-695
OBJECTIVETo construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase(-) Chinese hamster ovary cell (CHO-dhfr(-)) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western blotting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay.
RESULTSThe expression vectors stably expressed chimeric antibody in CHO-dhfr(-). In the cell supernatant of the F4' clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively.
CONCLUSIONThe human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.
Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; immunology ; pharmacology ; CHO Cells ; Cell Survival ; drug effects ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Mice ; Protein Engineering ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology
9.Protective effects of heat shock response on circulatory collapse induced by hyperthermia.
Bin WANG ; Bing-de LUO ; Fei ZOU ; Wei-ren WAN ; Jin-qiang GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(2):115-118
OBJECTIVETo investigate the protective effects and mechanism of heat shock response (HSR) on circulatory collapse induced by hyperthermia.
METHODSTwo experiments were carried out: (1) Protective effects of HSR. Rats were divided into 2 groups: heat shock (HS) group, sham control (SC) group. After HS group was pretreated with heat shock and recovered for 20 h at room temperature, both groups were exposed to heat till death, and blood pressure, electrocardiogram were measured continuously during exposure. Mean arterial pressure (MAP), survival time etc were acquired through Chart software. (2) Mechanism of effects. Rats were divided into 3 groups: HS group, SC group and normal control (NC) group. The treatment in HS and SC groups was identical with that in the first experiment, but it would be terminated at 73 min after heat exposure. Systolic pressure (Ps), diastolic pressure (Pd) etc were recorded and content of NO and HSP70 in myocardium were measured.
RESULTS(1) The survival time in HS group [(102.3 +/- 11.4) min] was longer than that in SC group [(87.9 +/- 7.7) min] and shock revealed later (P < 0.01); (2) During early heat exposure MAP in HS group was not different from that in SC group, but after 60 min MAP in HS group were higher than that in SC group; (3) MAP, Ps, Pd, HR and HSP70 in HS group were significantly higher but content of NO was lower than those in SC group (P < 0.01, P < 0.05).
CONCLUSIONHSR may induce upregulation of HSP70 and inhibit excessive production of NO in myocardium, thus result in relief of circulatory collapse induced by hyperthermia.
Animals ; Heat-Shock Proteins ; analysis ; Heat-Shock Response ; physiology ; Hot Temperature ; Male ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley ; Shock ; metabolism ; physiopathology ; Time Factors
10.Mechanism of copper and zinc on tanshinones of cultivated Salvia miltiorrhiza.
Bin WANG ; De-hui LIU ; Xian-he TAN ; Kang-cai WANG ; Ya-qin GUO ; Jing-yu DAI
China Journal of Chinese Materia Medica 2008;33(17):2082-2087
OBJECTIVETo study the mechanism of microelement including Cu and Zn on the accumulation of three danshinones in Salviae miltiorrhizae root and build a theory basis for its good quality and high yield.
METHODSand culture experiments were conducted to study the effect of Cu and Zn on the accumulation of three danshinones and oxidase including peroxidase and polyphenol oxidase activity in the plant root. The correlation between available Cu and Zn contents in matrix and oxidase activity in the plant root and, the correlation between available Cu and Zn contents in matrix and contents of tanshinones in the root were discussed.
RESULTContents of danshinones in the root increased with the increasing of Cu and Zn concentration. Dynamic monitoring on contents of dan-shinones of the plant roots growing in the pots with different Cu and Zn concentration in the whole growing season showed that the contents of danshinones for 60 days were the lowest, for 120 days the highest and then dropped for 150 days. In addition, among available Cu and Zn contents of matrix, oxidase including peroxidase and polyphenol oxidase activity and contents of tanshinones in the root,the correlation between two factors were significant difference (P < 0.05 or P < 0.01).
CONCLUSIONThe mechanism of Cu and Zn on the accumulation of danshinones may be that Cu and Zn improve the activity of peroxidase and polyphenol oxidase, which promote transformation of phenolic compounds to terpenes and therefore to increase contents of danshinones.
Catechol Oxidase ; metabolism ; Copper ; metabolism ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; analysis ; metabolism ; Peroxidase ; metabolism ; Phenanthrenes ; metabolism ; Plant Proteins ; metabolism ; Salvia miltiorrhiza ; enzymology ; metabolism ; Zinc ; metabolism