Genetic Association of Tumor Necrosis Factor-β, Interleukin-10, and Interleukin-1 Gene Cluster Polymorphism With Susceptibility to Pneumonia in Kidney Transplant Recipients

Introduction

Pneumonia remains a significant cause of morbidity and mortality after kidney transplantation. The present study was therefore conducted to investigate whether or not the polymorphisms of tumor necrosis factor (TNF)β, interleukin (IL)-10, IL-1β, and IL-1 receptor antagonist (IL-1ra) gene predicted the susceptibility to pneumonia within the first year after kidney transplantation.

Methods

Subjects comprised 33 kidney transplant recipients with pneumonia and 63 noninfected kidney transplant recipients. Genomic DNA from these 96 kidney transplant recipients was extracted from peripheral blood leukocytes. The regions containing the NcoI polymorphic site at position +252 of TNFβ gene, the RsaI polymorphic site at position −592 of IL-10 gene, and the AvaI polymorphic site at position −511 of IL-1β gene were amplified by polymerase chain reaction (PCR) and subsequently digested with NcoI, RsaI, and AvaI restriction enzyme, respectively. The polymorphic regions with intron 2 of the IL-1 ra gene (IL-1 RN) containing variable numbers of a tandem repeat of 86 base pairs, were amplified by PCR.

Results

Univariate analysis showed that recipient IL-10, IL-1β, and IL-1 RN polymorphisms were not associated with the presence of pneumonia (P = .589, .940, and .286, respectively). However, compared with GG genotype, recipient TNFβ +252AA + AG genotype was significantly associated with susceptibility to pneumonia (P = .006). Age of 45 years or older was not significantly associated with susceptibility to develop pneumonia but had a tendency to develop it (P = .119). After adjusting for age of 45 years or older, recipient TNFβ+252 AA + AG genotype (odds ratio = 5.366, 95% confidence intervals = 1.470 − 19.589, P = .011) independently predicted the risk for pneumonia within the first year after kidney transplantation in the multivariate analysis.

Conclusion

These results suggested that recipient TNFβ gene polymorphism may be useful in predicting pneumonia, hence identifying individuals who could benefit from preventive treatment and a less potent immunosuppression regimen.     

Effects of transforming growth factor-β1 on formation and activation of osteoclasts

Transforming growth factor-₁ (TGF-₁) has biological functions in various types of cells. However, its roles in the regulation of osteoclast formation and function are unclear. To examine them, we employed a culture system in which unfractionated cells obtained from long bones of 13-day-old mice were cultured on a dentine slice. We found that TGF-₁ has a potent inhibitory effect on osteoclastic bone resorption at a dose of 0.2–5 ng/ml. By electron microscopy the osteoclasts appeared to have fewer mitochondria and ruffled borders than those in control cultures. But in the presence of 1,25-dihydroxyvitamin D₃, [1,25-(OH)₂D₃], TGF-₁ at a dose of 0.2–1 ng/ml stimulated the formation of osteoclasts from unfractionated bone cell cultures in which preexistent osteoclasts had degenerated. Thus, using stromal cell-free he-mopoietic blast cells, we examined the direct action of TGF-₁ on osteoclast precursors. Although TGF-₁ inhibited tartrate-resistant acid phosphatase-positive (TRAP) multinucleate cell (MNC) formation induced by 1,25-(OH)₂D₃, the conditioned medium (CM) of TGF-₁-treated MC3T3-E1 cells stimulated such formation. These results suggest that TGF-₁ inhibits osteoclastic bone resorption but stimulates osteoclast formation via the action of factor(s) produced by TGF-₁-treated osteoblasts in the presence of 1,25-(OH)₂D₃.     

Expression of MCP-1 and TGF-β1 and its significance in early stage of obstructive jaundice in rats

Objective To investigate the expression of MCP-1 and TGF-β1 in early stage of ob-structive jaundice and explore its relation to liver correlated injury indexes in rats. Methods Fifty male Wistar rats were randomized into the control group, sham-operated group and biliary obstruction group. On day 10, serum ALT and BIL-T levels were determined in inferior caval blood and hepatic MDA was estimated in liver homogenates. Meanwhile, serum TGF-β1 level was measured by ELISA and the MCP-1 infiltration determined with immunohistochemistry. Results Serum ALT and BIL-T values were elevated. Meanwhile, the liver MDA content was increased. The positive expression of MCP-1 was augmented and serum TGF-β1 was over-expressed in the early stage after obstructive jaun-dice in rats. The hepatic MCP-1 expression had a positive correlation with the elevated ALT, BIL-T and MDA levels. However, the serum TGF-β1 content only had a positive correlation with ALT and BIL-T. Conclution Hepatic MCP-1 and serum TGF-β1 expression in the early stage after biliary tract obstruction is related to liver injury and extrahepatic cholestasis. Meanwhile, the hepatic MCP-1 ex-pression is correlated with hepatic oxidation stress. They have an important significance in liver injury in rats during the early period of obstructive jaundice.     

Early Expression of Hepatocyte Growth Factor, Interleukin-6, and Transforming Growth Factor-β1 and -β2 in Symptomatic Infection in Patients Who Have Undergone Liver Transplantation

Introduction

Early septic complications may be a deciding factor for successful recovery among patients who have undergone orthotopic liver transplantation. Therefore, monitoring liver function parameters plays an important role in postoperative treatment to achieve an early diagnosis of postsurgical complications. We ought to measure standard liver function parameters and the expression levels for selected cytokines among patients exhibiting symptoms of infection after orthotopic liver transplantation.

Materials and methods

The study was performed on 30 patients who were divided into two groups: SI-0 consisted of patients free of infection, and SI-1, those who had symptoms of infection. We determined standard liver function parameters and expression of hepatocyte growth factor (HGF), interleukin (IL)-6, transforming growth factor (TGF)-β1, and TGF-β2.

Results

There were no significant differences in standard liver function parameters between the two groups of patients. There were no significant differences in the levels of expression for the cytokines in question between the two groups of patients.

Conclusions

Although standard liver function parameters provide diagnostically valuable information on the patient's condition, they cannot be used to determine the extent of systemic infection among patients showing signs of infection after liver transplantation. Determining gene expression levels in circulating lymphocytes is a sensitive method to monitor patients' condition after liver transplantation. The expression levels of HGF, IL-6, TGF-β1, and TGF-β2 in circulating lymphocytes were not sufficiently specific to diagnose transitory postsurgical complications such as symptomatic infection.     

Role of Interleukin-1α and Type I Interleukin-1 Receptor in the Growth Activity and Invasion of Gastric Carcinoma Cells

To clarify the role of the interleukin (IL)-1/IL-1 receptor system in the progression of gastric carcinoma cells in patients with advanced gastric cancer, we measured the tissue concentrations of IL-1α, the expression of IL-1-receptor type I (IL-1RtI) on tumor cells, and the cell-growth activity through an analysis of DNA content. The concentrations of IL-1α were significantly higher in differentiated than in undifferentiated tumors (P = 0.038). The expression of IL-1RtI was upregulated in the tumor cells associated with IFNγ, scirrhous type tumors, and T3 and T4 tumors. There was a clear linear correlation between the tissue concentrations of IL-1α and S-phase fractions in differentiated tumors (r = 0.664, P = 0.003). Tumor cells with high IL-1α concentrations and low IL-1RtI expression had significantly greater S-phase fractions than those with low IL-1α concentrations, independent of IL-1RtI expression (P = 0.024 in low IL-1RtI, P = 0.019 in high IL-1RtI). These findings indicate that IL-1α stimulates the growth of differentiated gastric carcinoma cells and that IL-1RtI expression is involved in tumor invasive activity. High S-phase levels were not necessarily associated with a high expression of IL-1RtI, which may be due to the downregulatory effects of high IL-1α concentrations. Received: December 27, 2000 / Accepted: July 17, 2001     

Effects of low-level laser therapy on ROS homeostasis and expression of IGF-1 and TGF-β1 in skeletal muscle during the repair process

The aim of the present study was to determine the effects of low-level laser therapy (LLLT) on the homeostasis of reactive oxygen species (ROS) and expression of IGF-1 and TGF-β1 in the gastrocnemius muscles of rats following contusion. Muscle regeneration involves cell proliferation, migration, and differentiation and is regulated by growth factors. A growing body of evidence suggests that LLLT promotes skeletal muscle regeneration and accelerates tissue repair. Adult male Sprague-Dawley rats (n?=?96) were randomly divided into three groups: control group (no lesion, untreated, n?=?6), contusion group (n?=?48), and contusion-plus-LLLT group (n?=?42). Gallium aluminum arsenide (GaAlAs) laser irradiation (635 nm; beam spot, 0.4 cm²; output power, 7 mW; power density, 17.5 mW/cm²; 20 min) was administered to the gastrocnemius contusion for 20 min daily for 10 days. Muscle remodeling was evaluated at 0 h and 1, 2, 3, 7, 14, 21, and 28 days after injury. Hematoxylin and eosin and Van Gieson staining were used to evaluate regeneration and fibrosis; muscle superoxide dismutase (SOD) and malondialdehyde (MDA) were detected via biochemical methods; expression of transforming growth factor beta-1 (TGF-β1) and insulin-like growth factor-1 (IGF-1) were investigated via immunohistochemistry. The results showed that LLLT markedly promoted the regeneration of muscle and reduced scar formation. LLLT also significantly enhanced muscle SOD activity and significantly decreased muscle MDA levels 1, 2, and 3 days after injury. LLLT increased the expression of IGF-1 2, 3, and 7 days after injury and decreased the expression of IGF-1 21 and 28 days after injury. LLLT decreased the expression of TGF-β1 3 and 28 days after injury but increased expression at 7 and 14 days after injury. Our study showed that LLLT could modulate the homeostasis of ROS and of the growth factors IGF-1 and TGF-β1, which are known to play important roles in the repair process. This may constitute a new preventive approach to muscular fibrosis.     

Effects of IGF-Ⅱand TGF-β1 on invasiveness of placental trophoblast cells in patients with pregnancy-induced hypertension syndrome

Objective: This study was to investigate the invasiveness of pregnancy-induced hypertension (PIH) trophoblast cells and evaluate the effects of IGF-Ⅱand TGF-β1 on cytotrophoblast invasion.Methods: Cytotrophoblast cells from normal and PIH placenta were separated and purified. Cytotrophoblast invasiveness of normal and PIH placenta was measured by in vitro invasion assay. Effects of IGF-Ⅱand TGF-β1 on cytotrophoblast invasion were also studied.Results: In PIH group, cytotrophoblast invasiveness was dramatically decreased. In normal group, trophoblast invasiveness was significantly enhanced by IGF-Ⅱ but inhibited by TGF-β1. Neither IGF-Ⅱ nor TGF-β1 had statistically significant effects on PIH trophoblast invasion.Conclusions: PIH cytotrophoblast invasiveness dramatically decreases as compared to the normal level. IGF-Ⅱand TGF-β1 may play an important role in shallow trophoblast invasion on PIH.     

Effects of Ginsenoside Rb1 on proliferation of Schwann kcells in culture

Objective:To investigate the effects of Ginsenoside Rb1 on the proliferation ofSchwann cells in culture.Methods:Applying MTT assay and Thymidine incorporation assay,the effects of Ginsenoside Rb1 on the proliferation of Schwann cells isolated from the sciatic nerve of adult rat were studied.Results:Ginsenoside Rb1(10μg/ml)significantly induced Schwann cell proliferation,the effect was similar to NGF(50μg/ml).At high concentrations of Ginsenoside Rb1(1 mg/ml) ,the proliferation of Schwann cells was significantly inhibited.Conclusions:Ginsenoside Rb1 at the optimal concentratios is found to be effective in inducing the proliferation of Schwann cells ,but at higher concentrations the drug is cytotoxic for Schwann cells.     

Expression and significance of HIF-1α and ET-1 in gastrointestinal stromal tumors

Objective To explore the expression and the value of HIF-1α,and ET-1 in judging the prognosis of gastrointestinal stromal tumors (GISTs). Methods The expression of HIF-1α, and ET-1 protein was examined in 76 GISTs by immunohistochemistry S -P methods. Results There was a positive correlation between the expression of HIF-1 α and ET-1 ( P < 0.05 ). The positive expression rate of HIF-1 α and ET-1 was 73.68% (50/76) ,and 65.79% (50/76) respectively,which was related with histologicial grade, tumor diameter, infiltration and metastasis, nuclear division rating of GISTs ( P < 0.05 ), but had no relationship with patients' age, gender, initial position of the tumor ( P > 0.05 ). There was statistically sig-nificant difference in the expression of HIF-1 α and ET-1 in the following groups:among the three classes of very low-risk and low-risk, middle-risk, high-risk, bewteen the diameter < 2 cm and > 5 cm (P < 0.05). The more malignant degree and larger diameter, the more highly positive expression rate ( P < 0.05 ). The positive expression in the groups with infiltration and metastasis, and nuclear division ≥5/50 HP was sig-nificantly higher than the groups without infiltration and metastasis, and nuclear division < 5/50 HP (P < 0.05). Conclusion The expression of HIF-1α had a significant correlation with ET-1. HIF-1α,and ET-1 expression was closely related with the prognosis of GISTs,and can serve as important predictors for survival.     

Expression and significance of HIF-1α and ET-1 in gastrointestinal stromal tumors

Objective To explore the expression and the value of HIF-1α,and ET-1 in judging the prognosis of gastrointestinal stromal tumors (GISTs). Methods The expression of HIF-1α, and ET-1 protein was examined in 76 GISTs by immunohistochemistry S -P methods. Results There was a positive correlation between the expression of HIF-1 α and ET-1 ( P < 0.05 ). The positive expression rate of HIF-1 α and ET-1 was 73.68% (50/76) ,and 65.79% (50/76) respectively,which was related with histologicial grade, tumor diameter, infiltration and metastasis, nuclear division rating of GISTs ( P < 0.05 ), but had no relationship with patients' age, gender, initial position of the tumor ( P > 0.05 ). There was statistically sig-nificant difference in the expression of HIF-1 α and ET-1 in the following groups:among the three classes of very low-risk and low-risk, middle-risk, high-risk, bewteen the diameter < 2 cm and > 5 cm (P < 0.05). The more malignant degree and larger diameter, the more highly positive expression rate ( P < 0.05 ). The positive expression in the groups with infiltration and metastasis, and nuclear division ≥5/50 HP was sig-nificantly higher than the groups without infiltration and metastasis, and nuclear division < 5/50 HP (P < 0.05). Conclusion The expression of HIF-1α had a significant correlation with ET-1. HIF-1α,and ET-1 expression was closely related with the prognosis of GISTs,and can serve as important predictors for survival.     

Expression and significance of HIF-1α and ET-1 in gastrointestinal stromal tumors

Objective To explore the expression and the value of HIF-1α,and ET-1 in judging the prognosis of gastrointestinal stromal tumors (GISTs). Methods The expression of HIF-1α, and ET-1 protein was examined in 76 GISTs by immunohistochemistry S -P methods. Results There was a positive correlation between the expression of HIF-1 α and ET-1 ( P < 0.05 ). The positive expression rate of HIF-1 α and ET-1 was 73.68% (50/76) ,and 65.79% (50/76) respectively,which was related with histologicial grade, tumor diameter, infiltration and metastasis, nuclear division rating of GISTs ( P < 0.05 ), but had no relationship with patients' age, gender, initial position of the tumor ( P > 0.05 ). There was statistically sig-nificant difference in the expression of HIF-1 α and ET-1 in the following groups:among the three classes of very low-risk and low-risk, middle-risk, high-risk, bewteen the diameter < 2 cm and > 5 cm (P < 0.05). The more malignant degree and larger diameter, the more highly positive expression rate ( P < 0.05 ). The positive expression in the groups with infiltration and metastasis, and nuclear division ≥5/50 HP was sig-nificantly higher than the groups without infiltration and metastasis, and nuclear division < 5/50 HP (P < 0.05). Conclusion The expression of HIF-1α had a significant correlation with ET-1. HIF-1α,and ET-1 expression was closely related with the prognosis of GISTs,and can serve as important predictors for survival.