Comparative study of the effects of rhKGF, CBLB502 and WR2721 on radiation-induced oral mucositis / 解放军医学杂志

Objective To compare the protective effect of rhKGF, CBLB502 and WR2721 on radiation-induced oral mucositis (ROM). Methods Fifty male 6-8-week-old C57BL/6J mice were randomly divided into normal group, irradiation control group, rhKGF group, CBLB502 group, and WR2721 group (n=10 each). The 30-day survival rate and change in body weight of mice that had received 17Gy irradiation of head and neck area were recorded. In another group of 20 mice, 1% toluidine blue staining and HE staining were used to observe oral ulcers and pathological changes in the tongue tissue. The proliferation of keratinocyte cells was assessed by Ki-67 immunohistochemistry. Results Compared with the irradiation control group, administration of rhKGF and WR2721 could significantly improve the 30-day survival rate, accelerate the recovery of body weight, and promote the proliferation of keratinized epithelial cells of mice after irradiation, without inducing obvious oral mucositis. However, There was no significant difference between CBLB502 group and irradiation control group in survival rate, body weight and pathological changes in tongue tissues of mice. Conclusion rhKGF and WR2721 could alleviate ROM and improve the survival of mice, while CBLB502 has no such effect.

Human beta defensin-4 and keratinocyte growth factor gene expression in cultured keratinocyte and fibroblasts of burned patients

PURPOSE: To evaluate KGF and human beta defensin-4 (HBD-4) levels produced by dermic fibroblasts and keratinocytes cultivated from burned patients' skin samples. METHODS: Keratinocytes and fibroblasts of 10 patients (four major burns, four minor burns and two controls) were primarily cultivated according to standard methods. HBD-4 and KGF genes were analyzed by quantitative PCR. RESULTS: In fibroblasts, KGF gene expression was 220±80 and 33.33±6.67 (M±SD; N=4), respectively for major and minor burn groups. In keratinocytes, KGF gene expression was 11.2±1.9 and 3.45±0.37 (M±SD; N=4), respectively for major and minor burn groups. In fibroblasts, HBD-4 gene expression was 15.0±4.0 and 11.5±0.5 (M±SD; N=4), respectively for major and minor burn. In keratinocyte, HBD-4 gene expression was 0.0±0.0 and 13.4±4.8 (M±SD; N=4), respectively for major and minor burn. CONCLUSIONS: KGF expression was increased in burn patient fibroblasts compared to control group. In keratinocytes culture, KGF suppression is inversely proportional to burn extension; it is active and increased in major burn but decreased in minor burn. HBD-4 expression was increased in fibroblasts and decreased in keratinocytes from all burned patients. .

Keratinocyte growth factor, interleukins (1 beta, 6, 8, 10, 12), and tumor necrosis factor alpha in culture medium of dermal fibroblast of burned patients

PURPOSE: To evaluate the level of cytokines and keratinocyte growth factor (KGF) or Fibroblast Growth Factor 7 (FGF-7) in the culture medium of cultured human dermal fibroblasts from patients with large burn in comparison to small burn. METHODS: Fibroblasts of 10 patients (four large burns, four small burns and two controls) were initiated by the enzymatic method using collagenase. Cytokines and KGF in the supernatant of the culture medium was measured by, respectively, flow cytometry using Cytometric Bead Array Human Inflammation kit (CBA, BD Biosciences, USA) and the enzyme immunoassay method using the Quantikine (r) Human KGF. The experiments were performed in triplicate. RESULTS: The expression of IL-12 protein in patients with large burns showed a tendency to increase. IL- 6, IL- 10, and IL- 1beta were observed no difference. For IL - 8, TNF - alpha and KGF was observed a significant difference between the expression in large and small burned patient. CONCLUSION: That IL-8, TNF-alpha and KGF showed higher expression in cultured fibroblasts of large burned patients. .

Effects of histologic chorioamnionitis induced by lipopolysaccharide intra-amniotic injection on fetal lung maturation and development in preterm fetal rats / 中华围产医学杂志

Objective With various doses of lipopolysaccharide (LPS) intra-amniotic injection in pregnant rats to establish histologic chorioamnionitis (HCA) model,and to determine the effects of HCA on fetal lung maturation and development in preterm fetal rats.Methods Thirty pregnant Sprague-Dawley rats were randomly assigned to four groups.On 19 days of gestation,rats in three model groups (n=8) were given intra-amniotic injection of LPS 2,8,10 μg,respectively,and those in the only one control group (n=6)was given normal saline (NS) 40 μl.Fetal rats were taken out 48 hours after LPS injection.Placenta,fetal membrane,fetal lung and umbilical cord were collected for pathological examination.mRNAs of surfactant protein (SP)-A,B and C and keratinocyte growth factor (KGF) in lung were determined by quantitative real-time polymerase chain preaction.KGF protein expression in lung was measured by immunohistochemistry.Morphologic observation of lung was performed on postnatal day 3.R× C table Chi-square test,KruskalWallis test and analysis of variance were used as statistical methods.Results (1) Fetal rat mortality in control,LPS2,LPS8 and LPS10 group was 0.0％ (0/55),22.5％ (18/80),51.4％ (36/70) and 87.5％(35/40),respectively,which increased with increasing dose of LPS (x2=46.183,P＜0.005).(2) HCA,fetal lung inflammation and umbilical vasculitis were induced in LPS groups,and the severity was dosedependent (fetal lung inflammation:Hc=39.84,HCA:Hc=41.13,umbilical vasculitis:Hc=41.52,all P＜0.01).(3) The expression of SP-A,SP-B,SP-C and KGF mRNAs in the four groups (control,LPS2,8,10) had statistical differences (SP-A mRNA:0.99±0.28,2.48±0.34,1.09±0.31 and 0.09±0.09,F=78.051; SP-B mRNA:0.99±0.34,2.23±0.53,1.49±0.51 and 0.14±0.06,F=28.327; SP-C mRNA:1.20±0.39,2.00±0.20,1.04±0.37 and 0.12±0.16,F=39.546; KGF mRNA:0.97±0.19,2.18±0.61,0.93±0.09 and 0.21±0.11,F=37.544; all P＜0.01).All mRNA expressions in LPS2 group were higher than those in the control group and those in LPS10 group was lower (all P＜0.05).(4) The expression of KGF protein in LPS2 group was significantly higher than that in other groups (0.60±0.20 vs 0.28±0.12,0.37±0.22,0.24±0.12,F=17.280,all P＜0.01).(5) Alveolarization was significantly inhibited in LPS8 group on postnatal day 3,and less maturity of pulmonary tissue was observed.Conclusions Various doses of LPS intra-amniotic injection in rats could induce HCA,fetal lung inflammation and umbilical vasculitis with different degrees.Histological inflammation would be worse with increasing LPS dosage.Moderate inflammation could promote lung maturation without inducing bronchopulmonary dysplasia,and KGF may play a role in this process.

Effects and mechanisms of human umbilical cord-derived mesenchymal stem cells on the thymus development in nude mice / 中国组织工程研究

BACKGROUND:For autoimmune diseases, it is difficult to effectively solve the lack of immunological tolerance in patients by traditional treatments. Mesenchymal stem cells have the biological functions of tissue and organ regeneration and immune regulation. OBJECTIVE:To explore the effects and mechanisms of human umbilical cord-derived mesenchymal stem cells on the development of thymus in nude mice. METHODS:Human umbilical cord-derived mesenchymal stem cells were intraperitoneal y injected into BABL/c nude mice at a dose of 2×106 per mouse. We analyzed the maturation and distribution of thymic epithelial cells in the thymus rudiment of nude mice and the thymopoiesis of this newly developed thymus rudiment;furthermore, we explored the therapeutic effects of human umbilical cord-derived mesenchymal stem cells. RESULTS AND CONCLUSION:Our data showed wel-organized cortex-medul a structure and an obvious improvement in the maturation of thymic epithelial cells in the rudiment. We further demonstrated the improved thymopoiesis and the enhanced export of mature T cells with the T regulatory cells increase in peripheral blood. Furthermore, we found that human umbilical cord-derived mesenchymal stem cells could be engrafted in the thymus and express many cytokines, especial y keratinocyte growth factor which is essential for the thymus development. These findings indicate that a new mechanism of human umbilical cord-derived mesenchymal stem cells can provide a proper microenvironment for the reconstitution and functional maturation of thymus in nude mice, and elicit another insight in terms of therapeutic efficacy of human umbilical cord-derived mesenchymal stem cells in many autoimmune diseases.

Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients

PURPOSE: To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. METHODS: Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. RESULTS: The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). CONCLUSIONS: The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.