Objective To investigate the effects of the insulin-like growth factor 1 (IGF-1), the transforming growth factor β1(TGFβ1), and the basic fibroblast growth factor (bFGF) on proliferation and cell phenotype of the human fetal meniscal cells, and to find out the best combination and concentration of the growth factors for the meniscus tissue engineering. Methods The fetus came from the healthy woman accidental abortion and the procedure had got her approval.The human fetal meniscal fibrochondrocytes were cultured in vitro. The cell phenotype was identifiedby the collagen type Ⅱ immunohistochemistry and Aggrecan immunofluorescence. Inthe growth factor groups, the 3rd passage meniscal cells synchronized by the serum starvation method and were mixed with IGF-1 (1, 10, 50, 100 μg/L), TGF-β1 (0.1, 1.0, 5.0, 10.0, 50.0 μg/L), and bFGF (5, 10, 50, 100, 200 μg/L), respectively, and in the combination groups, the combinations of bFGF and TGF-β1, bFGF and IGF-1, TGF-β1 and IGF-1 were established at their optimal effect concentrations. The control group was also established for comparison. The dose-response relationship was studied at 48 h and 72 h bythe MTT colorimetric method. Results The 3rd passage meniscalcells could express collagen type Ⅱ and Aggrecan before and after the addition of the three growth factors. The proliferating effects of the growth factors (IGF-1 50 μg/L，TGF-β1 5 μg/L，bFGF 50 μg/L) on the 3rd passage cells at 48 h and 72 h were significantly better in the growth factor groups than in the control group (Plt;0.05)，and the combination groups of bFGF 50 μg/L and IGF-1 50 μg/L, IGF-1 50 μg/L and TGF-β1 5 μg/L showed a significantly higher proliferatingeffect than that in the single growth factor group (Plt;0.05). bFGF 50 μg/L and TGF-β1 5 μg/L had no synergetic effect (Pgt;0.05). Conclusion IGF-1, TGF-β1 and bFGF can promote the proliferation of the human fetal meniscal cells, respectively, and the combinations of bFGF and IGF-1, IGF-1 and TGF-β1 at their optimal concentrations can have better proliferating effects than the single growth factor. They can be used for the in vitro amplification of the meniscal seed cells.
Objective To investigate the relationship between the level of serum-insulin like growth factor-1( IGF-1) and the nut ritional status of cancerous cachexia. Methods Colon cancer CT-26 cells were implanted subcutaneously to 30 liver2specified IGF-1 gene deleted (L ID) C57BL/ 6 mice to establish cancerous cachexia model and theother 30 C57BL/ 6 mice were included as cont rol group. The serum levels of IGF-1 , cytokine TNF-αand IL-6 , bloodglucose , albumin and t riglyceride were detected respectively on day 14 , 18 and 22 af ter the plantation of tumor. Thebody weight of mice , tumor weight and the weight af ter tumor removed in two group s were measured respectively.Results Af ter the plantation , the levels of IGF-1 in L ID group at different times were all significantly lower thanthose in cont rol group ( Plt; 0. 05) . The serum levels of TNF-α, IL-6 , blood glucose and t riglyceride were ascendinggradually over time ( Plt; 0. 05) , but weight s af ter tumor removed and the level of albumin were descending in twogroup s ( Plt; 0. 05) . Compared with the cont rol group , the serum levels of IL-6 , TNF-α, blood glucose and t riglyceride in L ID tumor-bearing mice were all significantly higher at different time point s ( P lt; 0. 05) . On day 18 and 22 ,the weight s af ter tumor removed and the amount of ingestion in L ID group were significantly lower than those in thecont rol group ( Plt; 0. 05) . Conclusion Compared with the low level of IGF-1 in cancerous cachexia , normal level ofserum IGF-1 may represent lower degree of cancerous cachexia2related cytokines and better nut ritional state , whichmay provide a novel idea of the therapy of cancerous cachexia.
Objective To investigate the effect of short-term administration of growth hormone (GH) on serum insulin-like growth factor-1 (IGF-1) level and nutritional status in patients after gastrointestinal operation, and evaluate whether postoperative application of GH rise the risk of tumor recurrence. Methods Forty-eight patients undergoing major gastrointestinal operation were randomly divided into two groups: GH group （n=24） and control group （n=24）. The two groups received isocaloric isonitrogenous nutrition with daily injection of either GH 0.15 U/kg or placebo for a period of day 3-9 postoperatively. Serum albumin, fibronectin, and IGF-1 were measured before operation as a baseline, and day 3 and 10 after operation using standard laboratory techniques. Nitrogen balance was measured daily from day 3 to day 9 after operation. Postoperative complications and adverse reaction were observed. All cancer patients received regular abdominal B-type ultrasonography and chest X-ray examination during 2 years of follow-up. Results Compared with control group, GH treatment did not influence serum IGF-1 and serum albumin level （Pgt;0.05）, but improved significantly the rise from day 3 to day 10 of serum fibronectin level 〔(22.8±5.8) mg/L vs.（9.6±3.6） mg/L， P＜0.05〕 and the cumulative nitrogen balance 〔（11.37±16.82） g vs.（－9.11±17.52） g， P＜0.01〕 postoperatively. There was no severe adverse effects and complications during GH treatment. The tumor-recurrence rates were not statistically different between two groups during follow-up. Conclusions Short-term administration of low-dose GH combined with early nutrition support can improve total nitrogen retention and protein metabolism, but not influence serum IGF-1 level after major abdominal surgery. Short-term administration of low-dose GH may not cause the tumor-recurrence.
Objective To investigate the effects of insulin-like growth factor 1（IGF-1） and ethanol (EtOH) on the changes in the osteoblast proliferation and the osteoblast function under the normal serum concentration and serum starvationMethodsThe osteoblasts harvested from the SD rat calvaria were incubated in the following six conditions according to the supplements in DMEM: the F15group：15% newborn calf serum (NCS)； the F15/EtOH group：100 mmol/L of EtOH added to 15% NCS； the F2 group：2% NCS； the F2/EtOH group：100 mmol/L of EtOH added to 2% NCS；the F2/IGF-1 group：25ng/ml of IGF-1 added to 2% NCS；the F2/IGF-1/EtOH group：100 mmol/L EtOH added to 25 ng/ml IGF-1 and 2% NCS. The osteoblasts were analyzed by the MTTassay, alkaline phosphatase（ALP） activity, and RTPCR at 24, 48, 72 and 96h ours after the culture. Results The absorbance (A), the ALP activity, and the expression of BGP mRNA (the proliferation and function indicators of the osteoblasts) were significantly decreased in the F15/EtOH group at all the time points when compared with those in the F15the group (P＜ 0.05); the above 3 indicators were significantly decreased in the F2 groupwhen compared with those in the F15 group (P＜0.05); they were significantly decreased in the F2/EtOH group when compared with those in the F2 group (P＜0.05); however, the indicators in the F2/IGF-1 group were significantly increased when compared with those in the F2 group (P＜0.05); the A value in the F2/IGF-1/EtOH group was not significantly decreased when compared with that in the F2/IGF-1 group, with an exception of the A value at 24 hours (P＞0.05); however, ALP and BGP mRNA were significantly decreased (P＜0.05). All the indicators were significantly increased when compared with those in the F2/EtOH group (P＜0.05) Conclusion Ethanol can inhibit the osteoblast proliferation and the osteoblast function, and can increase the inhibition when the osteoblasts were cultured under the serum starvation. This may be one of the mechanisms for alcoholic bone disease. IGF-1 can prevent the inhibition of the osteoblasts under the serum starvation and counteract the ethanolinduced proliferation inhibition; therefore, IGF-1 is an alternaive therapeutic intervention for alcoholic bone disease.
ObjectiveTo evaluate the possible role of the expression of insulin-like growth factor-1 receptor (IGF-1R) in determining rectal cancer radiosensitivity. MethodsThe paired preradiation biopsy specimens and postoperative specimens were obtained from 87 patients with rectal cancer in the department of digestive tumor surgery, Jiangsu Province Hospital of Traditional Chinese Medicine, Affiliated Hospital of Nanjing University of Traditional Chinese Medicine from January 2009 to December 2010. The IGF-1R expression was examined by immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR). The tumor radiosensitivity was defined according to Rectal Cancer Regression Grade, then the relation between the IGF-1R expression and tumor radiosensitivity was evaluated. ResultsCompared with the preradiation biopsy specimens, IGF-1R expression significantly increased in the paired postoperative specimens of the residual cancer cells (Plt;0.001). The IHC result demonstrated IGF-1R overexpression was significantly associated with a poor response to radiotherapy (rs=0.401, Plt;0.001); RT-PCR detection of IGF-1R expression on preradiation biopsy specimens also showed that IGF-1R mRNA negative patients had a higher radiation sensitivity (rs=0.497, Plt;0.001). ConclusionDetection of IGF-1R expression may predict radiosensitivity of preoperative irradiation for rectal cancer.
Objective We investigated the effect of supplementation with alanyl-glutamine dipeptide on insulin resistance and outcome in patients with chronic obstructive pulmonary disease (COPD) and respiratory failure. Methods A prospective, randomized, open and controlled trial was conducted. Patients with COPD and respiratory failure were recruited between Jan 2005 to Feb 2006 and randomly assigned to a trial group (n=14) with glutamine dipeptide supplmented parenteral nutrition and a control group (n=16) with isocaloric, isonitrogenic parenteral nutrition. On the third day and fifth day of nutrition treatment, blood glucose was clamped at level of 4.4 to 6.1 mmol/L by intravenously bumped insulin. Blood gas, blood glucose level, insulin dosage were recorded everyday. The outcomes were mortality, length of stay (LOS) in hospital and in ICU, mechanical ventilation times and the costs of ICU and hospital.Results Thirty patients successfully completed the trial. There was no difference in blood gas between two groups, but PaO2 increased gradually. Compared with control group, blood glucose level had trend to decrease in trial group. The average insul in consumption decreased significantly in trial group on the fifth day. There was no statistical difference between two groups in mortality, length of stay in hospital and the costs of hospital. But compared with control group, length of stay in ICU and mechanical ventilation days had trend to decrease in trial group. Conclusion Alanyl-glutamine dipeptide do not improve pulmonary function of patients with COPD and respiratory failure. However, alanyl-glutamine dipeptide attenuated insul in resistance and stabilized blood glucose. This trial does not confirm alanyl-glutamine di peptide can improve outcome in critically ill patients with COPD and respiratory failure between two groups in mortality at the end of 30 days, length of stay in hospital and the costs of hospital. But the length of stay in ICU and the duration of mechanical ventilation does decrease, but not significantly, in the trial group.
ObjectiveTo observe the effect of lentivirus-mediated cyclooxygenase 2 (COX-2) and Aggrecanase-1 silencing and insulin-like growth factor 1 (IGF-1) in BMSCs after injecting into the knee joint cavity in cynomolgus monkeys with knee osteoarthritis (OA). MethodsBMSCs were isolated from the bone marrow of 10 donors. The lentivirus vector expressing genes of COX-2, Aggrecanase-1, and IGF-1 were constructed, and transfected into the third generation human BMSCs at 40 multiplicity of infection (virus group); BMSCs transfected with lentivirus-empty vector served as blank-virus group. The growth status and number of BMSCs were observed under inverted phase contrast microscope, and normal BMSCs were used as normal control group. At 1 week after transfected, the mRNA expressions of COX-2, Aggrecanase-1, and IGF-1 were detected with RT-PCR. Nine 3-year-old cynomolgus monkeys were selected to establish the OA model according to Hulth modeling method, and were randomly divided into 3 groups (n=3). At 6 weeks after remodeling, the right knee joint cavity was injected accordingly with 1 mL BMSCs (about 1×107 cells) in virus group and blank-virus group, with 1 mL of normal saline in the blank control group; the left knee served as normal controls. The general condition was observed after injection; at 1, 4, and 6 weeks, the concentrations of prostaglandin E2 (PGE2), IL-1, Aggrecanase-1, and IGF-1 of double knee liquid were detected with ELISA; at 6 weeks, MRI, general observation, histology method, and immunohistochemistry method were used to detect the knee cartilage changes and the expressions of COX-2, Aggrecanase-1, and IGF-1 were measured with RT-PCR. ResultsNo significant difference was found in cell morphology and growth curve between 2 groups after transfection. By RT-PCR, COX-2, and Aggrecanase-1 expressions were significantly reduced, IGF-1 expression was significantly increased in virus group when compared with normal control group and the blank-virus group (P < 0.05). All monkeys survived to the end of the experiment after injection. When compared with blank-virus group and blank control group, the concentrations of PGE2, Aggrecanase-1, and IL-1 significantly decreased and the concentration of IGF-1 significantly increased in the virus group (P < 0.05), but the indicators in 3 groups were significantly higher than those in the normal control group (P < 0.05). MRI showed that abnormal articular surface with high density could be found in virus group, blank-virus group, and blank control group, while the virus group had the minimum area. Gross observation and histological observation showed that the cartilage morphology of virus group, blank-virus group, and blank control group was accordance with early OA articular cartilage changes, but virus group was better than blank-virus group and blank control group in repair degree, whose improved Pineda score was significantly lower (P < 0.05). Immunohistochemical staining showed that the virus group had deeper dyeing with occasional brown particles and more chondrocytes than blank-virus group and blank control group. By RT-PCR, COX-2 and Aggrecanase-1 mRNA expressions of cartilage in virus group were significantly decreased, and IGF-1 expression was significantly increased when compared with blank control group and the blank-virus group (P < 0.05). ConclusionLentivirus-mediated multi-genes co-transfection in BMSCs can inhibit the expressions of COX-2 mRNA and Aggrecanase-1 mRNA, and enhance the IGF-1 mRNA expression, which decreases the concentration of inflammatory factors, and protects the joint cartilage effectively.
Objective To discuss the diagnosis and treatment of insulinoma, clinical characteristics and summarize our experiences. Methods The clinical, operative and pathologic findings from 15 cases of insulinoma of in our hospital from 1989 to 1998 were retrospectively studied. Results All 15 patients recieved surgical treatment. Fourteen patients were cured and the 15th patient died from hepatic failure six months later. Conclusion Insulinoma should be diagnosed and treated as early as possible. It can be cured by resection of the tumor.
Objective To identify the best therapy regimen for a patient with rare hypoglycemia due to insulin autoantibody (IAA). Methods We searched The Cochrane Library (Issue 3, 2008), PubMed (1966-July 2009), EMbase (1974-July 2009) and CBM (1978-July 2009) to identify relevant evidence. The quality of the retrieved studies was critically assessed. Results A total of 291 records were retrieved. No clinical guidelines, systematic reviews or clinical randomized studies were identified. Thirty treatment-related studies involving 6 interventions showed that insulin combined with Prednisone was relatively more effective and safer than conventional therapies. Conclusion The steroid treatment might be useful for the improvement of glycamic control in patients with high IAA levels and severe hypoglycemia and hyperglycemia due to insulin antibodies raised against subcutaneously-injected human insulin.
Objective To assess the tolerance of preoperative carbohydrate-rich beverage, to determine its effect on postoperative insulin resistance and analyze its potential mechanism. Methods Thirty-two patients undergoing elective colorectal cancer resection were recruited to this randomized controlled study and assigned to two groups at random. Patient in control group was fasted before operation, while patient in study group was given oral water. Homeostasis model assessment (HOMA) indexes, activity of PTK, and mRNA and (or) protein expressions of PKB, PI3K and GluT4 were measured before and (or) immediately after surgery. Furthermore preoperative well-beings of patients were studied. Results Among well-beings, feeling of thirst, hunger and anxiety tended to be better in patients receiving carbohydrate-rich beverages compared with fasted ones (P＜0.05). Whole body insulin sensitivity decreased by 33% in the study group while 38% in the control group (P=0.007 2), and the activity of PTK, expressions of PI3K and PKB in study group were higher than those in control group (P＜0.05， P＜0.01), but no significantly difference was observed about GluT4 in both groups (Pgt;0.05）. Conclusion Preoperative consumption of carbohydrate-containing fluids is safe and effective. Provision of carbohydrate energy source prior to surgery may attenuate immediate postoperative insulin resistance. A carbohydrate-rich drink enhances insulin action at the time of onset of anaesthesia or surgery by activating three kinases named PTK, PI3K, PKB which are key enzymes in pathway of insulin signal transduction. It is likely to explain the effects on postoperative insulin resistance.