Adult ; Antipsychotic Agents ; therapeutic use ; Diagnosis, Differential ; Humans ; Insulin-Like Growth Factor I ; biosynthesis ; Magnetic Resonance Imaging ; methods ; Male ; Mental Status Schedule ; Pituitary Neoplasms ; complications ; diagnosis ; Prolactinoma ; complications ; diagnosis ; Psychotic Disorders ; complications ; diagnosis

OBJECTIVETo optimize the culture media by adding the growth factors required to maintain tenocytes survival and promote their differentiation without fetal bovine serum (FBS) supplementation, in order for the approach to be used for any future tendon tissue engineering.

METHODSThe human tenocytes were cultured in α-MEM media by adding FBS at various concentrations and supplementing both insulin-like growth factor 1 (IGF-1) and transforming growth factor β-3 (TGFβ-3). A number of growth factors were selected that could support tenocytes expansion at reduced differentiated state with the minimum FBS. By employing fractional factorial design, different treatment groups went through AlamarBlue(TM) tests to evaluate the cell number growth whilst collagen quantification by real time RT-PCR technique and tenocyte differentiation were also studied.

RESULTSThe tenocytes cultured for 14 days with 0% FBS, 50 ng/ml IGF-1 and 10 ng/ml TGFβ-3 maintained survival over 14 days, the Cell count were 6228.68 ± 43.87. They were higher than the other experimental groups, but less than 10% FBS control group (13 576.74 ± 286.75, t = 41.29, P < 0.05). The tenocytes cultured in the treated group also showed enhanced collagen synthesis ((0.322 ± 0.003) ng, t = 4.13 - 5.93, P < 0.05).

CONCLUSIONThese findings have shown for the first time that human tenocytes could be maintained survival for a long period of time in the culture media without FBS, having this approach a suitable one for tendon tissue engineering.

Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Culture Media ; pharmacology ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Tendons ; cytology ; drug effects ; metabolism ; Transforming Growth Factor beta3 ; pharmacology

OBJECTIVETo examine the effects of high and low concentrations of transforming growth factor (TGF) β(1) and insulin-like growth factor-I (IGF-I) on the extracelluar matrix synthesis of the self-assembled constructs of temporomandibular joint (TMJ) disc.

METHODSThe experimental groups of self-assembled constructs were exposed to IGF-I (10, 100 µg/L) and TGF-β(1) (5, 50 µg/L), the control groups were not added with any growth factors. All groups were examined at 3 and 6 weeks for gross morphological, histological, and biochemical changes. Safranin-O/fast green staining was used to examine glycosaminoglycan (GAG) distribution, picrosirius red and immunohistochemical staining to observe type I collagen distribution. Type I collagen contents were tested by ELISA assay kit, GAG contents were measured by Blyscan GAG assay kit, and the cell numbers were quantified with a Picogreen reagent kit.

RESULTSThe growth factor groups all upregulated the matrix synthesis of the self-assembled constructs compared with control groups. TGF-β(1) (5 µg/L) and IGF-I (10 µg/L) were the two most potent concentration in increasing type I collagen and GAG synthesis and cells proliferation. IGF-I group (10 µg/L) produced nearly 2 times (109.16 ± 5.12 µg) as much type I collagen as the control group (69.13 ± 5.94 µg) at 3 weeks. The matrix contents and the number of the proliferated cells in control group and all GF groups at 6 weeks were more than those at 3 weeks.

CONCLUSIONSIGF-I (10 µg/L) is the most beneficial growth factor and can be applied in tissue-engineering stratigies of the temporomandibular joint disc. At the same time, the exposure time of growth factors is another key factor that affects matrix synthesis of TMJ disc constructs.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Extracellular Matrix ; metabolism ; Glycosaminoglycans ; biosynthesis ; Goats ; Insulin-Like Growth Factor I ; pharmacology ; Temporomandibular Joint Disc ; cytology ; metabolism ; Tissue Engineering ; methods ; Transforming Growth Factor beta1 ; pharmacology

Expression of zinc-finger protein 143 (ZNF143), a human homolog of the Xenopus transcriptional activator protein Staf, is induced by various DNA-damaging agents including etoposide, doxorubicin, and gamma-irradiation. ZNF143 binds to cisplatin-modified DNA, and its levels are increased in cancer cells that are resistant to anticancer drugs, including cisplatin, suggesting that it plays a role in carcinogenesis and cancer cell survival. However, the mechanism of ZNF143 induction in cancer cells remains unclear. Both insulin-like growth factor-1 (IGF-1) and its receptor (IGF-1R) have been reported to be overexpressed in cancer cells and to be related to anticancer drug resistance, but the identity of the relevant signaling mediators is still being investigated. In the present study, we observed that IGF-1 was able to induce ZNF143 expression in HCT116 human colon cancer cells and that wortmannin, an inhibitor of phosphatidylinositide 3-kinase (PI3-kinase), inhibited this induction, as did diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, and monodansylcardavarine (MDC), a receptor internalization inhibitor. Treatment with MDC decreased the IGF-1-stimulated generation of reactive oxygen species. Taken together, these data suggest that IGF-1 induces ZNF143 expression in cancer cells via PI3-kinase and reactive oxygen species generation during receptor internalization.
Antineoplastic Agents/pharmacology ; Cell Line, Tumor ; Cisplatin/pharmacology ; Colonic Neoplasms/enzymology/genetics/*metabolism ; HCT116 Cells ; Humans ; Insulin-Like Growth Factor I/*pharmacology ; Phosphatidylinositol 3-Kinase/*metabolism ; Reactive Oxygen Species/*metabolism ; Signal Transduction ; Trans-Activators/*biosynthesis/genetics

Alanine Transaminase ; blood ; Animals ; Antibodies ; pharmacology ; Apoptosis ; Collagen Type I ; biosynthesis ; DNA-Binding Proteins ; metabolism ; Disease Models, Animal ; Insulin-Like Growth Factor Binding Protein 1 ; immunology ; L-Lactate Dehydrogenase ; blood ; Liver ; drug effects ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; immunology ; metabolism ; Male ; Mice ; Protective Agents ; pharmacology ; Thioacetamide ; Transforming Growth Factor beta ; metabolism

This study was aimed to construct the recombinant adenovirus containing rat insulin-like growth factor 1(rIGF-1), and then to investigate its expression in islet beta-cells. RNA was extracted using Trizol from rat livers. rIGF-1 cDNA was obtained using RT-PCR. The purified RT-PCR products and pAdTrack-CMV were digested using Bg1 II and EcoR V and religated by T4 DNA ligase, then transformed into electro-competent JM109 bacteria and selected on Kanamycin LB plates. This plasmid pAd-CMV-rIGF-1 was linearized by PmeI and co-transformed into electro-competent BJ5183 bacteria with pAdEasy-1 and selected on Kanamycin LB plates. After having been screened, the extracted plasmid of positive bacteria was transfected into HEK 293 cells with liposome and was identified by the green fluorescence protein (GFP) expression. The recombinant adenovirus encoding rIGF-1 was named Ad-rIGF-1, and the viral particles were further amplified, purified, and its titer was about 4.0 x 10(8)pfu/ml. Ad-rIGF-1 was transfected into rat pancreatic beta cell lines- RINm5F cells, RT-PCR was carried out to detect the transfer genes, rIGF-1 protein in cells culture supernatants was detected by ELISA method, and its concentration was 91.6 +/- 26.8 ng/ml. rIGF-1 was present in Ad-rIGF-1-infected RINm5F cells as measured by Western blotting. The recombinant adenovirus vector containing rIGF-1 was constructed successfully, and the rIGF-1 protein was expressed by RINm5F cells. This method provided the mechanism of rLGF-1 to prevent beta cell from impairmentand to treat the case of type 1 diabetes.
Adenoviridae ; genetics ; metabolism ; Animals ; Cell Line ; Diabetes Mellitus, Type 1 ; therapy ; Genetic Therapy ; Genetic Vectors ; genetics ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Islets of Langerhans ; metabolism ; Rats ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Mechano-growth factor (MGF) is one of IGF-1 isoforms. MGF is mechanosensitive and has important functions in muscle hypertrophy, regeneration and nerve injury recovery. In this study, MGF cDNA (330 bp) was cloned from stretched osteoblasts by RT-PCR. In order to avoid prolin residue inhibiting enterokinase cleavage, 9bp of MGF cDNA 5' end sequence was truncated by primer, then the obtained truncated MGF (des(1-3)MGF) cDNA (321 bp) was subcloned in pET32a(+) vector to construct a prokaryotic recombination expression plasmid. Trx/des(1-3)MGF fusion protein, existing in forms of solution, was expressed in transformed Escherichia coli strain BL21(DE3) by IPTG induction at 30 degres C. The supernatant of cell lysates was subjected to ion exchange chromatography and Ni2+ metal affinity chromatography, and the fusion protein was obtained with the purity over 95%. After the fusion protein was cleaved by enterokinase, Trx and des(1-3)MGF was isolated by reverse-phase HPLC. Through these procedures, des(1-3) MGF was obtained with the purity of 98%. The protein molecular mass was conformity to the theoretical value by SDS-PAGE and mass spectrometry analysis. The purified des(1-3)MGF was incubated with MC3T3-E1 for cell proliferation and migration assays. The results show that des(1-3)MGF exhibited more facilitative effects on proliferation and migration of MC3T3-E1 than that of des(1-3)IGF-1.
Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Insulin-Like Growth Factor I ; Osteoblasts ; metabolism ; Protein Isoforms ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; STAT5 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo study the changes of insulin-like growth factor-I (IGF-I) in focal cerebral ischemical reperfusion injury model.

METHODSFocal cerebral ischemia was induced in rats using filament method, and the expressions of IGF-I was detected with immunohistochemical staining.

RESULTSThe expression of IGF-I was very low in normal brain tissues, but increased in the infracted area and penumbra after cerebral ischemia. The expressions of IGF-I reached the peak level of 14.83-/+0.48 at days 3 after reperfusion.

CONCLUSIONFocal cerebral ischemia reperfusion injury enhances the expression of endogenous IGF-I.

Animals ; Female ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; Insulin-Like Growth Factor I ; biosynthesis ; Ischemic Attack, Transient ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

BACKGROUND/AIMS: The insulin-like growth factor (IGF) system has been implicated in tumor growth, invasion, and metastasis. However, reports on the IGF-1 receptor (IGF-1R) based on radioimmunoassays are conflicting, and its prognostic implications in non-small-cell lung cancer (NSCLC) are still controversial. METHODS: Seventy-one paraffin-embedded tissue sections from stage I NSCLC patients were stained using a mouse monoclonal antibody against human IGF-1R. RESULTS: The intensity and frequency of IGF-1R expression on the membrane and cytoplasm of cancer cells was evaluated and scored using a semiquantitative system. IGF-1R expression was detected in nine of 71 (12.7%) cases. No significant relationship was found between clinical/histopathological parameters and IGF-1R expression. None of the patients whose tumor expressed IGF-1R had experienced distant metastasis or cancer-related death, although the difference did not reach statistical significance. CONCLUSIONS: We conclude that IGF-1R expression may not be a major prognostic factor for stage I NSCLC.
Adult ; Aged ; Aged, 80 and over ; Animals ; Carcinoma, Non-Small-Cell Lung/*immunology/mortality/pathology ; Female ; Humans ; Immunohistochemistry ; Insulin-Like Growth Factor I/*biosynthesis ; Male ; Mice ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; Receptor, IGF Type 1/*biosynthesis

With the use of a cyclic strain unit, the proliferation and gene expression of IGF-1 in the rat osteoblasts that underwent mechanical strain were studied. The cells were subjected to 15% elongation at frequency 20 cycles/min for different loading time. Under the action of different loading time, the relative proliferation index of the rat osteoblasts was the biggest of all when loading time was 12h; during the course, the expression of IGF-1 mRNA increased significantly, and then gradually tended toward 1 with the increase of the loading time. These results demonstrate that osteoblasts respond to the mechanical forces which may regulate the activities of osteoblasts indirectly by promoting the autocrine effect of IGF-1. Loaded osteoblasts can adjust and adapt themselves to new mechanical stimulation, and hence maintain a new state of equilibrium.
Animals ; Animals, Newborn ; Cell Proliferation ; Cells, Cultured ; Gene Expression ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Skull ; cytology ; Stress, Mechanical